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Food Emulsifiers and

Their Applications

Food Emulsifiers and


Their Applications
Edited by

Gerard L. Hasenhuettl
Kraft Foods

Richard W. Hartel
University of Wisconsin

SPRINGER-SCIENCE+BUSINESS MEDIA, B.V.

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12345678910 XXX 0100999897
Library of Congress Cataloging-in-Publication Data
Food emulsifiers and their applications/edited by Gerard L.
Hasenhuettl and Richard W. Hartel.
p. em.
Includes bibliographical references and index.
ISBN 978-1-4757-2662-6 (eBook)
ISBN 978-1-4757-2664-0
DOI 10.1007/978-1-4757-2662-6
1. Food additives. 2. Dispersing agents. I. Hassenhuettl, Gerard L., 1944II. Hartel, Richard W., 1951TP455.F667 1997
664'.06-dc21
97-1647
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To our wives and children

Contents

xiii

Contributors
Preface

XV

ONE
Overview of Food Emulsifiers

Gerard L. Hasenhuettl
1.1
1.2
1.3
1.4

Introduction
Emulsifiers as Food Additives
Emulsifier Structure
Emulsifier Functionality
References

1
2
5
7
9

TWO
Synthesis and Composition
of Food-Grade Emulsifiers

11

R. J. Zielinski
2.1
2.2
2.3
2.4
2.5

Introduction
Mono- and Diglycerides
Propylene Glycol Monoesters
Lactylated Esters
Polyglycerol Esters
vii

11
13
15
18
23

viii

Food Emulsifiers and Their Applications

2.6 Sorbitan Esters


2. 7 Ethoxylated Esters
2.8 Succinylated Esters
2.9 Fruit Acid Esters
2.10 Acetylated Monoglycerides
2.11 Phosphated Esters
2.12 Sucrose Esters
References

26
26
29
31
33
36
36
38

THREE
Analysis of Food Emulsifiers

39

Gerard L. Hasenhuettl

3.1
3.2
3.3
3.4
3.5
3.6

Introduction
Thin-Layer and Column Chromatography
Wet Chemical Analysis
Physical Methods
Instrumental Methods
Setting Specifications for Food Emulsifiers
References

39
40
43

52
54
62
63

FOUR

Carbohydrate/Emulsifier Interactions

67

Lynn B. Deffenbaugh

4.1 Introduction
4.2 Inclusion Complexes of Starch
4.3 Effects of External Lipid Materials on
Starch Properties
4.4 Factors Affecting Complex Formation
4.5 Physical Properties of Starch/Emulsifier Complexes
4.6 Summary
References

67
68
69
81
85
90
90

Contents

ix

FIVE

Protein/Emulsifier Interactions

Martin Bos, Tommy Nylander, Thomas Arnebrant,

95

and David C. Clark


5.1
5.2
5.3
5.4
5.5

Introduction
Protein Stability
Protein/Surfactant Interactions
Protein/Phospholipid Interactions
Protein/Emulsifier Interactions-Food Applications
References

95
97
98
120
132
137

SIX

Physicochemical Aspects of an Emulsifier


Functionality
147
Bjorn Bergenstahl
6.1
6.2
6.3
6.4

Introduction
Surface Activity
Solution Properties of Emulfiers
The Use of Phase Diagrams to
Understand Emulsifier Properties
6.5 Examples of the Relation between
Phase Diagrams and Emulsion Stability
6.6 Some Ways to Classify Emulsifiers
6.7 The Emulsifier Surface
References

147
148
149
153
154
161
167
171

SEVEN

Emulsifiers in Dairy Products


and Dairy Substitutes

173

Stephen R. Euston
7.1 Introduction
7.2 Ice Cream

173
174

Food Emulsifiers and Their Applications

7.3 Whipped Cream and Whipping Cream


7.4 Whipped Toppings
7.5 Cream Liqueurs
7.6 Creams and Coffee Whiteners
7. 7 Processed Cheese
7.8 Recombined, Concentrated and Evaporated Milks
7.9 Other Dairy Applications of Emulsifiers
7.10 Summary
References

183
187
191
194
197
199
202
203
204

EIGHT

Applications of Emulsifiers
in Baked Foods

211

Frank T. Orthoefer

8.1
8.2
8.3
8.4
8.5
8.6
8. 7
8.8

Introduction
History of Emulsified Shortenings
Emulsifier Function in Baked Goods
Role of the Shortening
Role of the Emulsifier
Emulsifier Interaction with Bakery Components
Applications in Baked Goods
Summary
References

211
211
213
215
216
221
225
233
234

NINE

Emulsifiers in Confectionery

235

Mark Weyland

9.1 Introduction
9.2 Emulsifiers in Chocolate and Compound Coatings
9.3 Antibloom Agents in Chocolate and
Compound Coatings
9.4 Other Emulsifiers Used in Coatings
9.5 Emulsifiers in Nonchocolate Confectionery

235
236
244
248
249

Contents

xi
252
253

9.6 Processing Aids


References

TEN
Margarines and Spreads

255

Eric Flack

10.1
10.2
10.3
10.4
10.5
10.6
10.7
10.8
10.9
10.10
10.11
10.12

Introduction
Early Development
Yellow-Fat Consumption
Definitions and Descriptions
Structure and Raw Materials
Fat Crystallization
Emulsifiers
Processing
Reduced- and Low-Fat Spreads
Oil-in-Water Spreads
Liquid Magarine
Summary
References

255
256
257
258
262
263
267
270
274
278
278
279
279

ELEVEN
Emulsifier Trends for the Future

281

Gerard L. Hasenhuettl

Globalization in the Food Industry


Nutritionally Driven Changes in Food
Trends toward Safer Emulsifers
Emulsifer Structure and Interactions
with Other Ingredients
11.5 Enzymatic Synthesis of Food Emulsifiers
References

11.1
11.2
11.3
11.4

Index

282
282
283
284
285
286
287

Contributors

Thomas Arnebrant
Department of Pharmaceutical Analysis
Ferring AB
Box 30047
S-216 13 Malmo, Sweden
Bjorn Bergenstahl
Institute for Surface Chemistry
P.O. Box 5607
S-114 86 Stockholm, Sweden
Martin Bos
TNO Nutrition & Food Research Institute
P.O. Box 360
3700 AJ Zeist, The Netherlands
David C. Clark
DMV International
NBC-Iaan 80
P.O. Box 13
5460 BA Veghal, The Netherlands

xiii

xiv

Contributors

Lynn B. Deffenbaugh
Hill's Pet Nutrition
Science & Technology Center
P.O. Bo.c 1658
Topeka, KS 66601-1658
Stephen R. Euston
New Zealand Dairy Research Institute
Private Bag 11029
Palmerston North, New Zealand
Eric Flack
Danisco Ingredients, Ltd.
North Way
Bury St. Edmonds
Suffolk, 1P32 6NP, United Kingdom
Gerard L. Hasenhuettl
Kraft Foods, Inc.
801 Waukegan Rd.
Glenview, IL 60025
Tommy Nylander
University of Lund, Chemical Center
Department of Physical Chemistry
P.O. Box 124
S-22100 Lund, Sweden
Frank T. Orthoefer
2004 Me Cracken St.
Stuttgart, AR 72160
Mark Weyland
Quest International
5115 Sedge Blvd.
Hoffman Estates, IL 60192
Richard J. Zielinski
Quest International
Joliet Technical Center
24708 Durke Dr.
Joliet, IL 60410-5249

Preface

Food emulsions have existed since long before people began to process foods for
distribution and consumption. Milk, for example, is a natural emulsion/colloid in
which a nutritional fat is stabilized by a milk-fat-globule membrane. Early
processed foods were developed when people began to explore the art of cuisine.
Butter and gravies were early foods used to enhance flavors and aid in cooking.
By contrast, food emulsifiers have only recently been recognized for their ability to stabilize foods during processing and distribution. As economies of scale
emerged, pressures for higher quality and extension of shelf life prodded the development of food emulsifiers and their adjunct technologies. Natural emulsifiers,
such as egg and milk proteins and phospholipids, were the first to be generally
utilized. Development of technologies for processing oils, such as refining,
bleaching, and hydrogenation, led to the design of synthetic food emulsifiers.
Formulation of food emulsions has, until recently, been practiced more as an
art than a science. The complexity offood systems has been the barrier to fundamental understanding. Scientists have long studied emulsions using pure water,
hydrocarbon, and surfactant, but food systems, by contrast, are typically a complex mixture of carbohydrate, lipid, protein, salts, and acid. Other surface-active
ingredients, such as proteins and phospholipids, can demonstrate either synerXV

xvi

Preface

gistic or deleterious functionality during processing or in the finished food.


The formulator of food emulsions has therefore traditionally been an experienced individual who reasoned by analogy to obtain desired technical effects
in the new food products. Recent impressive progress has been made in understanding the physical chemistry of food emulsions, dispersions, and foams by
application of sophisticated instrumentation and computing power. An appreciation of ingredient interactions has also been developed, as many of the references in this book bear testimony. However, a coherent work focusing on the
design of emulsifiers for food applications has been notably absent.
In this volume, we have attempted to collect material that clarifies the process
of designing a commercially viable emulsifier system for new products or improvement of existing foods. The process begins with an understanding of the
role and possibilities of the emulsifier to not only stabilize emulsions but also to
provide critical secondary functionalities. Manufacturing technology is described and analytical tests that ensure the quality of the emulsifier ingredient
are presented. Interactions of food emulsifiers with carbohydrates, proteins, and
water are significant in their use and are extensively discussed. Applications of
emulsifiers in the dairy, bakery, confectionery, and margarine industries demonstrate the reasoning process used to develop emulsion-based products. It is our

hope that this effort will stimulate further innovation directed at increasing the
quality and reducing the expense of processed food products.

Acknowledgments
The editors express their apprec1atwn to Barbara Bagnuolo and Julie
Hasenhuettl for typing and proofreading large portions of the manuscript.

Gerard L. Hasenhuettl
Richard W. Hartel

Food Emulsifiers and


Their Applications

ONE

Overview of Food Emulsifiers


Gerard L. Hasenhuettl

1.1

Introduction

Food emulsions, colloids, and foams have their origins in the evolution of animal species. Milk has a naturally occurring membrane that allows the dispersion of fat droplets into an aqueous environment. Early food formulations to
produce butter, whipped cream, cheese, and ice cream built upon the natural
emulsifiers present in the system. The development of mayonnaise in France
as a cold sauce utilized the natural egg phospholipids to disperse a liquid oil
into an acidified aqueous phase. The emulsifying power is still very impressive
by today's standards since it allowed up to 80% oil to be dispersed without inversion to an oil-continuous emulsion. The invention of margarine by the
French chemist Hippolyte Mege-Mouries in 1889 utilized the solid fat of tallow to produce a stable oil-continuous emulsion to serve as a low-cost substitute for butter. In this case, the emulsion had to be stable temporarily only
until the product was chilled.
Synthetic emulsifiers have come into common use only in the latter half of
the twentieth century. Their development was driven by the emergence of the
processed food industry, which needed technology to produce and preserve
quality products that could be distributed through mass market channels. A
key technical hurdle was to maintain product stability over an extended shelf

Food Emulsifiers and Their Applications

life. With small amounts of emulsifier, for example, salad dressings can be
stored on a shelf for more than a year without visible separation. Now, other
factors such as the development of rancidity limit consumer acceptance of aging products.
Detailed knowledge of the physical chemistry of emulsions is best obtained
when pure oil, water, and surfactants are used. Food emulsions, by contrast, are
extraordinarily complex systems. Commercial fats and oils are complex mixtures
of triglycerides that also contain small amounts of highly surface-active materials (Friberg and Larsson, 1990). Salt content and pH in food emulsions vary
widely enough to have significant effects on their stability. Natural and commercial emulsifiers are often complex mixtures that vary in composition between different manufacturers. Other food ingredients, such as proteins and particulates,
contribute surface activity that may dramatically alter the character of the emulsion. Because of all these complex relationships, the formulation of food emulsions grew up as an art, dominated by individuals having a great deal of
experience. The gradual development of sophisticated techniques such as electron microscopy, nuclear magnetic resonance, and chromatography/mass spectrometry has solidified the art with a scientific dimension.
The subject of food emulsions has been extensively reviewed by Friberg
and Larsson (1990), Krog et al. (1985), and Jaynes (1985). This book will be
oriented to the design, manufacture, and use of food emulsifiers as ingredients
to improve the quality and economy of processed foods.

1.2

Emulsifiers as Food Additives

Approximately 500,000 metric tons of emulsifiers are produced and sold


worldwide. However, since the value/volume ratio of these products is low,
very little truly global trade has yet developed. In the United States, food
emulsifiers fall into two categories: substances affirmed as GRAS (21CFR184)
and direct food additives (21CFR172). Substances that have been affirmed as
GRAS (generally recognized as safe) usually have less stringent regulations attached to their use. However, Food and Drug Administration Standards of
Identity may preclude their use in certain standardized foods. In comparison,
direct food additives may be allowed only in certain specific foods at low maximum allowable levels. The method of manufacture and analytical constants
may also be defined. The most commonly used food emulsifiers are listed in
Tables 1.1 and 1.2.

Overview of Food Emulsifiers

Table 1.1 Emulsifiers affirmed as GRAS


Emulsifier

2ICFR
No.

Lecithin

Functionalities

Typical
applications

184.1400

Coemulsifier, viscosity
reducer

Margarines, chocolate
products

Monoglycerides

184.1505

Emulsifier, aerator,
crystal stabilizer

Margarines, whipped
toppings, peanut
butter stabilizers

Diacetyltartaric acid
esters of
monoglycerides

184.1101

Emulsifier, film former

Baked goods,
confections, dairy
product analogs

Monosodium salt of
analogs, phosphated
monoglycerides

184.1521

Emulsifier, lubricant,
release agent

Dairy products,
soft candy

The European Economic Community (EEC) has also developed regulations


for food additives that may be utilized within the member nations. Substances
are divided into four annexes that have some resemblance to United States
regulations. Annex I is similar to the GRAS list (2ICFRI82). These additives
may be used anywhere except in natural or standardized processed foods.
Annexes 2 and 3 resemble direct food additive regulations. A separate listing
for solvents and solubilizing agents is contained in Annex 4, which is not separated in FDA regulations. Although the additive regulations are similar, great
care must be taken in trade between the United States and the EEC because
some specific emulsifiers are defined differently. For example, emulsifiers derived from polyglycerols up to decaglycerol are permitted in the United States,
but they are limited to tetraglycerol or lower in the EEC.
Other parts of the world have not developed into trading blocks with defined
regulations. Each country may have a unique perspective on which emulsifiers
may be allowed in food products. The problem is compounded by the fact that
regulations are written in the national language, requiring extensive and careful
translation. Some major differences in technology may develop because of a
country's unique specifications. For example, polysorbates have not been permitted as food additives in Japan. As a result, technology for production of
higher cost polyglycerol and sucrose esters has been widely developed there.

Food Emulsifiers and Their Applications

Table 1.2 Emulsifiers classified as direct food additives


Emulsifier

21CFR
No.

Functionalities

Typical
applications

Emulsifier, plasticizer,
surface-active agent

whipped toppings

Lactylated
monoglycerides

172.850

Acetylated
monoglycerides

172.828

Film former, moisture


barrier

Fruits, nuts, pizza

Succinylated
monoglycerides

172.830

Emulsifier, dough
strengthener

Shortenings, bread

Ethoxylated
monoglycerides

172.834

Emulsifier, stabilizer

Cakes, whipped
toppings, frozen
desserts

Sorbitan
monostearate

172.842

Emulsifier, rehydrating
agent

Confectionery
coatings, yeast,
cakes, icings

Polysorbates

172.836
172.838
172.840

Emulsifier, opacifier,
solubilizer, wetting agent

Salad dressings,
coffee whiteners,
gelatins, ice cream

Polyglycerol esters

172.854

Emulsifier, aerator, cloud


inhibitor

Icings, salad oils,


peanut butter,
fillings

Sucrose esters of
fatty acids

172.859

Emulsifier, texturizer,
film former

Baked goods, fruit


coatings,
confectionery

Calcium and sodium


Stearoyllactylates

172.844
172.846

Emulsifier, dough
conditioner, whipping
agent

Bread, coffee
whiteners, icings,
dehydrated

Baked products,

potatoes
Propylene glycol
esters

172.858

Emulsifier, aerator

Cake mixes,
whipped toppings

Overview of Food Emulsifiers

As with any other totally new food additive, the need to prove safety of the
product in foods at high levels of consumption requires extensive toxicity studies
and enormous documentation. The consequent financial and time commitments
make development of totally new emulsifiers unattractive for emulsifier manufacturers. A somewhat easier development approach is to petition for expanded
use (new applications or higher permitted levels) of emulsifiers that are already
approved. However, even this tactic may require several years of review.
In addition to national regulations, many food processors require their ingredients, including food emulsifiers, to be kosher so that their products are
acceptable to Jewish and many Islamic consumers. For emulsifiers to be
kosher, they must be produced from kosher-certified raw materials. This requirement precludes the use of almost all animal fats. This is not much of a
problem since emulsifiers are easily produced from vegetable fats that can be
blended to give similar fatty acid compositions. The major concern in kosher
certification is to determine in advance whether the customer's rabbinical
council recognizes the Hekhsher (kosher symbol) of the producer's rabbi.
Products labeled as "all natural" must contain ingredients that have not
been chemically processed or modified. Only lecithin or other naturally occurring materials such as proteins would be acceptable for these products.

1.3

Emulsifier Structure

Surface-active compounds (surfactants) operate through a hydrophilic head


group that is attracted to the aqueous phase, and an often larger lipophilic tail
that prefers to be in the oil phase. The surfactant therefore positions itself at
the air/water or oil/water interface where it can act to lower surface or interfacial tension, respectively. Figure l.l shows some typical hydrophilic and
lipophilic groups. Lipophilic tails are composed of Cl6 (palmitic) or longer
fatty acids. Shorter chains, such as Cl2 (lauric), even though they can be excellent emulsifiers, can hydrolyze to give soapy or other undesirable flavors.
Unsaturated fatty acids are Cl8 molecules having one (oleic) or two (linoleic)
double bonds. Linoleic acid is usually avoided since it is easily oxidized and
may produce an oxidized rancid off-flavor in the finished food. Fats may be hydrogenated to produce a mixture of saturated and unsaturated fatty acids.
Emulsifiers produced from these fatty acids may have an intermediate consistency (often referred to as "plastic") between liquid and solid. These products

Food Emulsifiers and Their Applications

Lipophilic end

Hydrophilic end
-OH

Saturated; palmitate or stearate

Unsaturated; oleate

Unsaturated; linoleate

Figure 1.1

Typical hydrophilic and lipophilic groups.

also contain measurable concentrations of trans (E) fatty acids that have higher
melting points than the cis (Z) fatty acids.
Polar head groups may be present in a variety of functional groups. They
may be incorporated to produce anionic, cationic, amphoteric, or nonionic surfactants. Mono- and diglycerides, which contain an -OH functional group, are
the most widely used nonionic emulsifiers. Lecithin, whose head group is a
mixture of phosphatides, may be visualized as amphoteric or cationic, depending on the pH of the product.
Proteins may also be surface active due to the occurrence of lipophilic
amino acids such as phenylalanine, leucine, and isoleucine. lnterfacially active proteins will fold so that lipophilic groups penetrate into the oil droplet
while hydrophilic portions of the chain extend into the aqueous phase.
Proteins in this configuration may produce a looped structure that provides
steric hindrance to oil-droplet flocculation and coalescence. Charged proteins
may also stabilize emulsions due to repulsion of like charged droplets.
Proteins may also destabilize water-in-oil emulsions, such as reduced fat margarines, by causing the emulsion to invert.
Food emulsifiers may be thought of as designer molecules because the
structure and number of heads and tails may be independently varied. A very
useful conceptual tool is hydrophile/lipophile balance (HLB). The topic has
been extensively reviewed by Becher, so only a brief description will be presented here. The number and relative polarity of polar groups in a surface-ac-

Overview of Food Emulsifiers

tive molecule determine whether the molecule will be water- or oil-soluble (or
-dispersible). This concept has been quantitated by calculation of an HLB
value to describe a given emulsifier. High-HLB values are associated with
easy water dispersibility. Since conventional practice is to disperse the surfactant into the continuous phase, high-HLB emulsifiers are useful for preparing
and stabilizing oil-in-water (0/W) emulsions. Low-HLB emulsifiers are useful
for formulation of water-in-oil (W/0) emulsions, such as margarine. Extreme
high or low values are not functional as emulsifiers since almost all of the molecule will be solubilized in the continuous phase. They would, however, be
very useful for full solubilization of another ingredient, such as a flavor oil or
vitamin, in the continuous phase. At some intermediate values of HLB, the
molecule may not be stable in either phase and will result in high concentration at the interface.

1.4 Emulsifier Functionality


In addition to their major function of producing and stabilizing emulsions, food
emulsifiers contribute to numerous other functional roles, as shown in Table
1.3. Some foods, notably chocolate and peanut butter, are actually dispersions

Table 1.3 Other functions offood emulsifiers


Emulsifier frmction

Example(s)

Whipping (aerating) agent


Dispersant
Dough conditioner
Defoamer
Starch complexer
Crystallization inhibitor
Antistaling agent
Antis ticking agent
Antispattering agent
Freeze-thaw stabilizer

Whipped toppings
Flavors, vitamins
Bread, buns, rolls
Yeast and sugar manufacturing
Macaroni, pasta
Salad oil
Yeast-raised baked goods
Candy, chewing gum
Margarines, frying shortenings
Frozen toppings and coffee

Gloss enhancer
Cloud formers
Hydrating agents
Encapsulating agents
Dispersion stabilizers

Whiteners
Confectionery coatings
Beverages
Powdered milk drinks
Flavors, aromas
Peanut butter

Food Emulsifiers and Their Applications

of solid particles in a continuous fat or oil phase. Chocolate viscosity is controlled by the addition of soy lecithin or polyglycerol ricinoleate (PGPR). Oil
separation in peanut butter is prevented by use of a monoglyceride or highmelting fat. In some cases the secondary effect may be of greater concern than
formation of the emulsion. Strengthening of dough and retardation of staling
are vital considerations to processors who bake bread.
A common practice in the food industry is to use two- or three-component
emulsifier blends to achieve multiple functionalities. In a cake emulsion, for
example, aeration to produce high volume, foam stabilization, softness, and
moisture retention are achieved by using an emulsifier blend. One useful statistical method to optimize emulsifier blends is the full factorial experimental
design using a zero or low level of each emulsifier and a higher level of each
emulsifier. The major advantage of this design is that it will detect two- and
three-factor interactions that are not uncommon in complex food systems.
Small-molecule emulsifiers (e.g., monoglycerides) may exert their effect by
partially or totally displacing proteins from an oil/water interface. This replacement is entropically favored because of the difference in size and mobility of the species. Direct interaction of emulsifiers and proteins may be
visualized through electrostatic and hydrogen bonding, although it is difficult
to observe in a system that contains appreciable amounts of oil. Chapter 5 on
emulsifier/protein interactions will elaborate on these concepts.
Emulsifier suppliers generally employ knowledgeable technical service
professionals to support their customers product development efforts. Their experience in selecting emulsifiers for a functional response is a valuable initial
source of information. However, food processors may want to develop unique
products that have no close relationship to a product currently in commerce. In
this case, the supplier may have some general ideas for emulsifier selection.
However, it may be necessary for product developers to define their own criteria for emulsifiers based on critical functions required in the product.
Statistical experimental design is a very useful tool to optimize food emulsifiers and their concentrations. For example, a full factorial design (Krog et al.,
1985; Jaynes, 1985) may be used to determine the levels of three emulsifiers to
obtain an optimum product. Response surface methodology (RSM) and fractional factorial designs are very useful techniques because they reduce the
number of experiments necessary to obtain optimal concentrations. However,
since synergistic and antisynergistic effects are often observed between emulsifiers, care should be taken to design the experiments so that two-factor inter-

Overview of Food Emulsifiers

actions are not confounded. Robust design is also highly recommended so that
the food product has minimal sensitivity to uncontrolled noise factors.
The objective of this book is to provide the food industry professional or interested technical professional with an overview of what emulsifiers are, how they
are prepared, and how they are utilized in food products. Although in many
senses food emulsifiers have become commodity ingredients, sophisticated understanding and application in processed foods is likely to continue to advance.

References
Friberg, S., Larsson, K. (eds.) (1990). Food Emulsions, Marcel Dekker, New York.
Jaynes, E. {1985). "Applications in the food industry, II," in Encyclopedia of Emulsion
Technology {ed. P. Becher), Vol. 2, Marcel Dekker, New York, 1985, pp. 367--85.
Krog, N., Rilson, T.H., Larsson, K. (1985). "Applications in the food industry, " in
Encyclopedia of Emulsion Technology (ed. P. Becher), Vol. 2, Marcel Dekker, New
York, pp. 321-66.

TWO
Synthesis and Composition of
Food-Grade Emulsifiers
R.J. Zielinski

There are a number of families of food-grade emulsifiers, which may be classified as


Mono- and diglycerides
Propylene glycol monoesters
Lactylated esters
Polyglycerol esters
Sorbitan esters
Ethoxylated esters
Succinylated esters
Fruit acid esters
Acetylated mono- and diglycerides
Phosphated mono- and diglycerides
Sucrose esters
In this chapter the general methods of preparing these food-grade emulsifiers
and some of the characteristics of each group of esters will be discussed.

2.1

Introduction

The food-grade emulsifiers are generally esters composed of a hydrophilic (water-loving) end and a lipophilic (fat-loving) end. In general, the lipophilic end
is composed of stearic, palmitic, oleic, or linoleic acid or combinations of these
fatty acids. The hydrophilic end is generally composed of hydroxyl or carboxyl

11

12

Food Emulsifiers and Their Applications

groups. The melting point of the various esters within each family will be determined by the melting point of the fatty acids used to prepare the emulsifier.
The melting points of the common fatty acids are given in Table 2.1. When
stearic and palmitic acids dominate, the ester will be solid and relatively highTable 2.1 Melting points of fatty acids

C8:0
CIO:O
Cl4:0
Cl6:0
Cl8:0
Cl8:1
Cl8:2

Acid

CMP

Caprylic acid
Capric acid
Lauric acid
Myristic acid
Palmitic acid
Stearic acid
Oleic acid
Linoleic acid

16.3
31.5
44.2
56.4
62.9
69.6
14
-6

eC)

melting; when oleic and linoleic acids dominate, the ester will be low-melting
and could be a liquid at room temperature. The fatty acids present in an emulsifier may be obtained from either a fat or oil or a fatty acid source. All fats and
oils are triglycerides, and the fatty acids can be obtained from the triglycerides
by a hydrolytic process followed by fractional distillation. Generally, either
natural or fully hydrogenated fats and oils are split to obtain the fatty acids.
This process is illustrated in Equation 1.
0

II

CH 2-0-C-R

CH 2-QH

II

CH-o--c-R1

CH 2-0-C-R

3H 20

CH-QH

CH 2-0H

R--cOOH

R1--cOOH

R2--cooH

0
Triglyceride

Glycerine

Fatty acids

(R = saturated or unsaturated alkyl chain)


(1)

where R, R 1, and R2 are the alkyl portion of fatty acid groups.

13

Synthesis and Composition of Food-Grade Emulsifiers

Kosher oleic acid may be obtained from high-oleic safflower and high-oleic
sunflower seed oils. It may also be obtained from a specially purified tall oil
from pine trees. Commercial stearic acid may be of three types: (i) a mixture of
about 90% stearic acid and 10% palmitic acid, (ii) a mixture of about 70%
stearic acid, 30% palmitic acid, or (iii) a mixture of about 50% palmitic acid
and 50% stearic acid; all are known as stearic acid. Generally, partially hydrogenated fats and oils are used to prepare plastic-type emulsifiers.

2.2

Mono- and Diglycerides

The mono- and diglycerides are the most widely used food-grade emulsifiers.
They may be esters that are solid and high-melting, esters that are liquid at
about room temperature, or a plastic-type ester. There are a variety of monoand diglycerides commercially available at the present time. These are commonly designated as (i) 40% a-monoglycerides, (ii) 50% a-monoglycerides,
and (iii) 90% monoglycerides. Monoglycerides are commonly composed of a
number of components, as indicated in Figure 2.1.
These components are present in different amounts in the commercially
available esters. The various types of commercial compositions are described
in the following discussion.
The mono- and diglycerides may be prepared by either an interesterification process with glycerine and triglycerides or a direct esterification process
with fatty acids. These processes are illustrated in Equation 2. In the latter
case, water of esterification is formed and removed during the process step.

0
CH 2-0H

CH-OH

CH-C-R

CH-OH

II

CH 2-0-C-R

II

CH-0-C-R

II

CH2-0-C-R

II

CH-0-C-R

II

CH 2-0H

CH-0-H

CH 2-0H

CH2-0-C-R

Glycerine

Monoglyceride

Diglyceride

Triglyceride

Figure 2.1

14

Food Emulsifiers and Their Applications

II

CH 2-0H

CH 2-0-c-R

CH-QH

II

heat

CH-0-C-R

CH-OH

CH 2-0H

CH-OH
catalyst

II

CH-QH
0

II

CH 2-0H

CH 2-0-C-R

CH 2-0H

CH-Q-C-R

Glycerine

Triglyceride

Glycerine

Monoglyceride

CH-OH + R-cOOH

I
Glycerine

II

CH 2-0H

CH 2-0H
heat
catalyst

CH 2-D-c-R

II

CH-O-c-R

II

II

CH-0-C-R

II

CH 2-0-C-R

CH 2-D-C-R

Diglyceride

Triglyceride

(2)

The catalysts commonly used in these processes are sodium hydroxide or hydrated lime (calcium hydroxide); the temperatures involved range from 200 to
250C. The proportions of each of the products-free glycerine, monoglyceride,
diglyceride, and triglyceride-are purely dependent on the molar ratio of glycerine and oil or glycerine and fatty acid used. The overall composition can be approximated using a random distribution of the free hydroxyl group and fatty acid
groups (Feuge and Bailey, 1946). The various fatty acid groups from the oil or
fatty acid are also assumed to he randomly distributed in the final product.
Once the desired ratio of oil or fatty acid and glycerine has been chosen to
yield the desired monoglyceride content, a number of different types of monoglyceride compositions are still potentially available. For example, if a 40%
monoglyceride composition is desired, then (i) the catalyst may be neutralized,

Synthesis and Composition of Food-Grade Emulsifiers

15

generally by phosphoric acid; (ii) the catalyst may be not neutralized; (iii) the
reaction mixture may be cooled to a determined temperature and the glycerine
that is insoluble in the final composition removed by decantation type process;
and (iv) the excess free glycerine may be removed from the reaction products
by vacuum distillation. A 40% monoglyceride prepared by a decantation-type
process (iii) may contain about 4% free glycerine, while a 40% monoglyceride
prepared by a vacuum-stripping process will typically contain less than 1%
free glycerine. A stripped 40% monoglyceride will typically be composed of
about 46% monoglyceride, 43% diglyceride, 10% triglyceride, and 1% glycerine. Similarly, a 50% monoglyceride may be prepared by decantation from insoluble glycerine or by a vacuum-stripping process. A vacuum-stripped 50%
monoglyceride will typically contain about 55% monoglyceride, 38% diglyceride, 5% triglyceride, and 2% free glycerine.
Under the conditions of high temperature, very low pressure, and an extremely short path, monoglycerides may be distilled and thus concentrated and
purified. This process is known as "molecular distillation." Typically, a 40%
monoglyceride mixture is subjected to molecular distillation to yield monoglycerides of over 90% purity. The nondistilled portion is recycled for an additional interesterification process to yield another 40% monoglyceride-type
composition for use as the feedstock.
As previously mentioned, either nonhydrogenated or partially hydrogenated
triglycerides, or saturated or unsaturated fatty acids, may be used to prepare
solid, plastic, or liquid monoglyceride mixtures.
The mono- and diglycerides have a generally recognized as safe (GRAS)
FDA status.

2.3

Propylene Glycol Monoesters

Propylene glycol or 1,2-propanediol is a food-allowed, dihydroxy polyol that is


used to prepare a variety of food-grade emulsifiers. The 1,3-propanediol is not
allowed in preparing food-grade emulsifiers. There are two methods of preparing food-grade propylene glycol monoesters (PGME): (i) interesterification of
propylene glycol with triglycerides and (ii) direct esterification with fatty acids.
In contrast to the mono- and diglycerides, both procedures do not yield the
same compositions. When an interesterification process is used, the final composition contains mono-, di-, and triglycerides in addition to propylene glycol
mono and diesters. This is illustrated in Equation 3.

16

Food Emulsifiers and Their Applications

II

CH 2-QH

CH-OH

CH 2-0-c-R

heat

CH-O-C-R

catalyst

II

CH 3

CH 2-0-c-R

Propylene
glycol

Triglyceride

0
CH 2-QH

CH-QH

II

II

CH 2-o-c-R

CH-OH

CH 2-0-c-R

CH-0-C-R

CH2-0H

CH-OH

CH 3

CH 3

CH 3

CH2-QH

Propylene
glycol

Propylene
glycol
monoester

Propylene
glycol
diester

Glycerine

II

CH 2-0-c-R

CH 2-Q-C-R

CH-QH

II

CH 2-o-c-R

CH-0-C-R

II

CH-o-c-R

CH-QH

CH 2-QH

CH 2-o-c-R

Monoglyceride

Diglyceride

Triglyceride

(R =saturated or unsaturated alkyl chain)


(3)

Synthesis and Composition of Food-Grade Emulsifiers

17

Basic catalysts such as sodium hydroxide or hydrated lime (calcium hydroxide)


are used in the interesterification process. Generally, the excess propylene glycol
and the glycerine formed during the reaction are removed by a vacuum distillation
process. The basic catalyst is neutralized prior to the distillation process to prevent
disproportionation of the products during the distillation process; the most commonly used acid is 85% phosphoric acid. The composition of the final product is
controlled by the molar ratio of propylene glycol to triglyceride in the starting reaction mixture. The approximate final concentrations of propylene glycol monoester
and monoglyceride may be determined by assuming that a random distribution occurs during the reaction process (Feuge and Bailey, 1946). By controlling the molar ratio of propylene glycol to triglyceride, propylene glycol monoester contents
ranging from about 13 to 70% may be obtained. Generally, most commercial compositions contain 50 to 70% propylene glycol monoester.
Commercial products containing propylene glycol monoesters of greater
than 90% are produced by means of a molecular distillation process similar to
that employed for the production of distilled monoglycerides. In the PGME
case, an interesterification mixture containing a high concentration of PGME
is used as a feedstock for the distillation process. Small amounts of distilled
monoglycerides may co-distill with the propylene glycol monoester.
The direct esterification of propylene glycol with fatty acids yields a mixture of free propylene glycol, propylene glycol monoester, and propylene glycol
diester, as illustrated in Equation 4.
The residual propylene glycol is generally removed by a vacuum steamstripping process. The amount of propylene glycol monoester ranges from 45
to 70% in commercially available compositions prepared by direct esterification of propylene glycol with fatty acids. The amount of monoester can be
approximated by assuming a random distribution with various molar ratios of
propylene glycol to fatty acid (Feuge and Bailey, 1946). A basic catalyst
such as sodium hydroxide or hydrated lime is generally used during the esterification step to avoid the formation of dipropylene glycol esters. The basic catalyst is neutralized with an acid, usually 85% phosphoric acid, prior
to the steam-stripping step to avoid a disproportionation of the monoester
during the stripping step.

If a strong acid is used as a catalyst during the direct esterification step,


then the distinct possibility of the self-condensation of propylene glycol to the
dimer or trimer followed by esterification of the dimer or trimer exists. Thus,
acid-catalyzed esterifications can contain dipropylene glycol monoesters and

18

Food Emulsifiers and Their Applications

CH 2--o-c-R

CH 2-0H

CH-OH

R-cOOH

heat

CH-OH

catalyst

CH 2

CH 3

II

CH 2-0-c-R

CH 2-0-C-R

I
(4)

tripropylene glycol monoesters. These esters are not allowed by the FDA as direct food additives.
The FDA regulations for propylene glycol monoesters allow the use of all
edible oils and edible fatty acids. However, the majority of the propylene glycol monoesters commercially available contain very high percentages of
palmitic acid and stearic acid that are saturated fatty acids. Few unsaturated
propylene glycol monoesters are available at this time.

2.4

Lactylated Esters

Lactic acid, 2-hydroxypropanoic acid, is a bifunctional acid. As illustrated in


Figure 2.2, fatty acid esters may be prepared by reaction with either the hydroxyl group or the fatty acid group. Lactic acid can also self-react to form
polymeric (dimer, trimer, etc.) chains. These polymers can also react with fatty
acid moieties. This is also illustrated in Figure 2.2.
This self-condensation reaction cannot be avoided, and all lactic acid
emulsifiers contain mixtures of monomer, dimer, and trimer esters.

Synthesis and Composition of Food-Grade Emulsifiers

19

/R-COOH
OH f--- R-COOH

OH

I 0II

CH 3-c-COOH

f--- R-QH

CH 3-C-c-o-cOOH

f--- R-OH

Lactic acid

CH3

Dilactic acid

Figure2.2

2.4.1

Reaction at the Hydroxyl Group

Two broad categories of food-grade emulsifiers that are prepared by the reaction
of the hydroxyl group of lactic acid with either fatty acids or fatty acid chlorides
are commercially available: (i) the lactylic esters of fatty acids and (ii) partial
metal salts of lactylic esters of fatty acids. Each will be discussed in tum.
2.4.1.1

Lactylic &ters of Fatty Acids.

Two types of lactylic esters of fatty acids

are commercially available. One composition contains approximately 60%


monomer, 10% dimer and trimer, 30% free fatty acid, and 3% free polylactic
acid. The second composition contains approximately 42% monomer, 16%
dimer and trimer, 30% free fatty acid, and 12% polylactic acid. Three methods
are available to prepare a lactylic ester with a 60% monomer content. The first is
the reaction of lactic acid of known monomer content with a fatty acid chloride
with or without the presence of a weak base to capture the hydrogen chloride
generated (Thompson and Buddemeyer, 1956). This method uses a costly acid
chloride and an extensive purification process. The second method involves the
direct esterification of about a 1:1 molar ratio of sodium lactate and fatty acid,
followed by acidification with mineral acid and isolation of the fatty lactylic ester
(Eng, 1972). A third method of preparing a lactylic ester with a 60% monomer
content is to react lactic acid with acetic anhydride to form the lactyl acetate.
The lactyl acetate is then subjected to an acidolysis reaction with fatty acids in
the presence of a sodium base under high-vacuum conditions. The mixture is
then acidified with mineral acid and the fatty lactylic esters isolated.
The lactylic ester with about 42% monomer and 16% dimer and trimer

20

Food Emulsifiers and Their Applications

acids most probably is prepared by the direct esterification of lactic acid with
fatty acid in about a 1:1 molar ratio (Thompson et al., 1956). A high free polylactic acid content (see Chapter 3 for analytical procedure) indicates that the
product has not been water-washed.
There are no restrictions on the lactic acid content of the lactylic esters of
fatty acids in the FDA regulations (21CFR172). There are no FDA restrictions
on the edible fatty acids that may be used in the preparation of the lactylic esters of fatty acids; however, the stearate esters are most commonly produced.
2.4.1.2

Metallic Salts of Lactylic Esters of Fatty Acids. There are two metallic salts of lactylic esters of fatty acids that are used in the food industry: calcium salt and sodium salt. The calcium salt was commercialized first, as a
dough conditioner for bread, and is known as calcium stearoyl-2-lactylate.
The 2 indicates that 2 moles of lactic acid were used in its preparation. The
sodium salt, which was commercialized later, is known as sodium stearoyl-2lactylate, sodium stearoyllactate, or simply "SSL." It is used extensively in
many food applications.
Both esters can be prepared by the same process, the direct esterification of
the partial salt of lactic acid with a fatty acid in about a 2: l molar ratio of lactic acid to fatty acid (Thompson et al., 1956). Both esters have a monomer content of about 40%, a polymeric content of about 31%, and a polylactic acid
content of about 6%.
Care must be taken to use high-quality starting materials to avoid excessive
darkening during the reaction process. The color of the final product can be reduced by the use of hydrogen peroxide as a bleaching agent.
The amounts of lactic acid and metal ion content of both esters are regulated
by the FDA and are about 34% and 4.5%, respectively. Only the use of specified
grades of stearic acid is permitted by the FDA regulations for the products.

2.4.2
2.4.2.1

Reaction at the Carboxylic Acid Group


Lactylated Monoglycerides.

The carboxylic acid group of lactic acid

can react with the hydroxyl group of other fatty acid-derived compositions to
yield emulsifiers that are useful in many food products. One of the most common
fatty acid compositions used is a reaction product with a monoglyceride to form a
lactylated monoglyceride identified in the FDA regulations as glycerol-lacto esters of fatty acids. The overall simplified reaction is illustrated in Equation 5.
As can be seen from Equation 5, the number of hydroxyl groups remain the

Synthesis and Composition of Food-Grade Emulsifiers

21

OH

I
+ CH -C-COOH
3
I
H

____.

r,--o--c

OH

I I

-c-cH,

+ Hp

CH-OH

I
Monoglyceride

Lactic acid

Lactylated monoglyceride

(5)

same in the reaction product as in the starting monoglyceride and a methyl


group is introduced. This has the net effect of lowering the melting point and
increasing the fat solubility of the lactylated monoglyceride compared to the
starting monoglyceride.
As previously discussed, lactic acid can homopolymerize; therefore, dilactate and trilactate esters of monoglycerides can and will be formed during the
esterification reaction.
A major variable in the formation of lactylated monoglycerides is the composition of the monoglyceride that is reacted with lactic acid. For example, Barsky
(1950) disclosed the use of shortening compositions containing lactylated monoglyceride made by two different methods. The first lactylated monoglyceride was
prepared by the reaction of l mole of oleic acid with l mole of glycerine; then the
reaction product was reacted with 33% of 88% lactic acid. The second monoglyceride was prepared by the interesterification of 100 parts of partially hydrogenated oil with 25 parts of glycerine; then the reaction product was reacted with
29.5% of anhydrous lactic acid. In both cases, about a nominal 40% monoglyceride intermediate composition was made, and about 7% of free glycerine could
be expected to be present in the monoglyceride mixture. The free glycerine can
react with lactic acid to yield water-soluble, oil-insoluble, glycerol lactates.

22

Food Emulsifiers and Their Applications

These glycerol lactates can be removed by water-washing or can be steamstripped from the reaction mixture under reduced pressure.
A distilled monoglyceride could also be used as a reacting monoglyceride
composition. However, during the reaction with lactic acid, the distilled monoglyceride could totally or partially disproportionate to yield free glycerine and
a mixture of mono-, di-, and triglycerides. Thus, the benefit of using a distilled
monoglyceride could be lost.
Other possibilities of starting monoglycerides are an unstripped intermediate monoglyceride with an a-mono content of 28 to 40%, a 40% U-monoglyceride that has been vacuum-stripped to remove the free glycerine, or a 50%
monoglyceride type that has or has not been vacuum-stripped to remove the
free glycerine before reaction with lactic acid. Of course, any diglycerides present in the monoglyceride mixture can also react with lactic acid.
Another variable in the production of lactylated monoglycerides is the
amount of lactic acid reacted with the monoglyceride intermediate. In theory,
a monoglyceride can react with 2 moles of lactic acid to yield the dilactylated monoglyceride ester. Commercially, lactylated monoglycerides are
available that contain either about 15% or 22% lactic acid. There is an analytical constant known as WICLA (see Chapter 3), which stands for water-insoluble combined lactic acid, used by some companies to characterize
lactylated monoglycerides. This value will give an estimation of the amount
of active functional lactylated monoglyceride present in the commercial
product as opposed to a total lactic acid value that is a summation of the
WICLA value and the amount of lactic acid present in a water-soluble form,
such as glycerol lactates.
Because of the wide range of possible starting monoglycerides and the reaction conditions chosen, all commerciallactylated monoglyceride compositions
may not be compositionally the same, and possibly, differences in functionality
can exist between products with a similar lactic acid content.
The FDA regulations for lactylated mono- and diglycerides permit the use
of any edible fat or oil or edible fatty acids to be used in their manufacture.
There are no restrictions on the amount of lactic acid that can be present in the
lactylated mono- and diglycerides.
2.4.2.2

Lactylated Fatty Acid Esters of Glycerol and Propylene Glycol.

Another

emulsifier, prepared from lactic acid by the reaction of the carboxylic acid
group of the lactic acid with the hydroxyl groups of glycerol and propylene gly-

Synthesis and Composition of Food-Grade Emulsifiers

23

col monoesters, is the lactylated fatty acid esters of glycerol and propylene glycol. In this type of emulsifier, propylene glycol is interesterified with an edible
fat or oil to yield a mixture of mono- and diglycerides and propylene glycol monoesters. The reaction mixture is then reacted with lactic acid. As with the
lactylated mono- and diglycerides, a great variety of propylene glycol monoester and monoglyceride mixtures could be used to react with lactic acid to
yield a final composition. In addition, lactic acid can react with any free propylene glycol and free glycerine initially present or formed by a disproportionation of the starting propylene glycol monoesterlmonoglyceride composition.
These propylene glycol lactates and glycerol lactates are removed by waterwashing or vacuum distillation of the reaction product. Obviously a complex
composition is produced.

It should be noted that only propylene glycol esters prepared by an interesterification process may be used. Propylene glycol mono- and diesters prepared by a direct fatty acid esterification route are not allowed under the
pertinent FDA regulation (21CFR172.850). The FDA regulation further specifies the WICLA content of the final ester composition to be 14 to 18%.

2.5

Polyglycerol Esters

Polyglycerol esters have been commercially available to the food industry for
over 25 years, but they have been known in the nonfood industry for many
more years. The polyglycerol alcohols are most often prepared by the polymerization of glycerine with an alkaline catalyst at elevated temperatures
(Harris, 1941; Babayan and Lehman, 1973). The polymerization is a random
process and a number of different polyglycerols are produced. This is illustrated in Equation 6.
The extent of polymerization is followed by refractive index, viscosity, or
hydroxyl value. When the theoretical hydroxyl value for a diglycerol is obtained, the polyglycerol could be called diglycerol. When the hydroxyl value
for triglycerol is obtained, the polyglycerol could be called a triglycerol, etc.
The hydroxyl values for di- to decaglycerol are summarized in Table 2.2. The
theoretical molecular weight of each polyglycerol is also given in Table 2.2.
In general, no effort is made to separate the various polyglycyerols, and the
entire reaction mixture that contains a distribution of polyglycerols is used to
prepare polyglycerol esters. At lower degrees of polymerization, higher concentrations of lower poyglycerols are present; at higher degrees of polymeriza-

24

Food Emulsifiers and Their Applications

heat
catalyst

OH

OH

OH

I I

OH

OH

OH

I I

OH

OH

OH

OH

CH 2-cH-cH 2-0-CH 2-CH-CH 2-0H

OH

Diglycerol

CH 2-CH-(H 2-0-CH 2-(H-(H 2-0-cH 2-CH-CH 2-0H

OH

I I

Triglycerol

CH 2-CH-cH 2-G-CH 2-CH-CH 2-0-CH 2-CH-CH 2-0-CH 2-CH-CH 2-0H


Tetragl ycerol

and so on, up to decaglycerol + (n- 1) H20


(6)
Table 2.2
Polyol
Glycerine
Diglycerol
Triglycerol
Tetraglycerol
Pentaglycerol
Hexaglycerol
Heptaglycerol
Octaglycerol
Nonaglycerol
Decaglycerol

Theoretical
hydroxyl value

Theoretical
molecular weight

1839
1352
1169

92
166
240
314
338
462
536
610
684
758

1072

1012
971
942
920
902
888

Synthesis and Composition of Food-Grade Emulsifiers

25

tion, higher concentrations of higher polyglycerols are present. The number of


possible esterification sites in a polyglycerol is the nominal polyglycerol plus
2(n + 2). Thus, a triglycerol has 5 possible esterification sites and an octaglycerol has lO possible esterification sites.
The polyglycerol esters may be prepared from the polyglycerols by either a
direct esterification with fatty acids or an interesterification with triglycerides.
When a fatty acid process is used, the theoretical molecular weight of the
polyglycerol is used with the molecular weight of the fatty acids to calculate
the reaction charge. Generally, if low degrees of esterification are used, the reaction product is neutralized and allowed to "phase out" at about l00C. The
free polyol is separated and the emulsifier is filtered and packaged. If higher
degrees of esterification are used, then no polyol will phase out and the product, neutralized or not neutralized, is filtered and packaged.

If an interesterification process is used, then additional glycerine from the


fat or oil is introduced into the reaction mixture. This additional glycerine will
modify the polyol distribution of the final product compared to the polyol distribution of the starting polyglycerol. Additional mono- and diglycerides will
be produced compared to a direct esterification process, and distribution of
fatty acids ester will be different than if a direct esterification process were
used. Esters prepared by an interesterification process are still identified by
the polyol that was initially used.
It is obvious that even if a single fatty acid were used to make polyglycerol
esters that the number of compositions possible are very numerous and com-

plex. Since mixtures of fatty acids are used, this makes the polyglycerol esters even more complex. Thus, a triglycerol monostearate polyglycerol ester
is a mixture of the stearate and palmitate esters of glycerine, diglycerol,
triglycerol, tetraglycerol, pentaglycerol, hexaglycerol, heptaglycerol, octaglycerol, nonaglycerol, and decaglycerol, while a decaglycerol tristearate is
a mixture of the stearate and palmitate esters of the same polyglycerols but in
different proportions.
A wide range of polyglycerol esters are commercially available from liquid
esters such as hexaglycerol dioleate to plastic esters like triglycerol monoshortening to solid esters like decaglycerol decastearate.
FDA regulations permit the use of edible hydrogenated or nonhydrogenated nonlauric oils and the edible fatty acids derived from them as well
as oleic acid from tall oil to prepare polyglycerol esters up to and including
decaglycerol.

26

Food Emulsifiers and Their Applications

2.6

Sorbitan Esters

Only one sorbitan ester is currently approved in the FDA regulations as a direct food additive in the United States: sorbitan monostearate. However, a
GRAS petition has been filed to permit the use of sorbitan tristearate in confectionery coatings (Federal Register, 1988). The FDA, as of 1996, has not
acted on this petition. The most commonly used process for the production of
sorbitan esters is the direct fatty acid esterification of sorbitol with stearic acid
(Brown, 1943; Japanese Patent, 1974). Generally, the stearic acid used is
about a 50:50 mixture of stearic and palmitic acids. The overall reaction is
shown in Equation 7.
Sorbitan is a monoanhydro sorbitol and sorbide (or isosorbide) is a dianhydro sorbitol, as illustrated in Equation 7. Commercial sorbitan monostearate is
the stearate, palmitate ester of a mixture of about 1 to 12% sorbitol, 65 to 72%
sorbitan, and 16 to 32% isosorbide. It can be demonstrated that the amount of
sorbitan is fairly constant but that the amount of linear sorbitol and isosorbide
can vary among manufacturers.
The use of an acidic catalyst during the esterification of sorbitol promotes
the cyclization of sorbitol to the mono- and dianhydro forms. The use of a basic
catalyst promotes the esterification reaction at the expense of color formation.
It is possible that each commercial supplier has its own blend of acidic and
basic catalysts to generate the ratio of sorbitan esters required to meet tight
FDA specifications for hydroxyl number, saponification number, and acid
number while minimizing color degradation of the product. Hydrogen peroxide
can be used to reduce the color of a sorbitan monostearate.
Stockburger (1981) has patented a process for preparing sorbitan esters in
which sorbitol is first dehydrated with an acid catalyst to the desired ratio of
sorbitol, sorbitan, and sorbide polyols. The mixture of polyols is then reacted
with fatty acids and a basic catalyst to yield the desired sorbitan ester with an
improved light color. The use of sorbitan monostearate in foods is highly regulated. The reader is referred to 21CFR172.842 for further details on where sorbitan monostearate can be used in food products, and for the required
analytical constants.

2.7

Ethoxylated Esters

Four ethoxylated fatty acid esters have FDA approval for use as direct food additives. These are ethoxylated sorbitan monostearate, ethoxylated sorbitan

Synthesis and Composition of Food-Grade Emulsifiers

CH 2-0H

CH 2-DH

CH-OH

CH-OH

CH-OH

27

catalyst

CH-OH

R-COOH

CH-OH

CH-OH

CH-DH

CH-OH

II

CH 2-0H

CH 2-0-C-R

Sorbitol

Linear sorbitol
monoester

J=l

CHI

L<JH
I

r-<JH
I
I

r_j

I
o

CH_j

CH-OH

CH

CH-OH

CH 2-0-C-R

Sorbitan

Sorbide

monoester

monoester

(7)

28

Food Emulsifiers and Their Applications

monooleate, ethoxylated sorbitan tristearate, and ethoxylated saturated monoand diglycerides. The base ester that has been ethoxylated is sorbitan monostearate, sorbitan monooleate, sorbitan tristearate, and a 28% monoglyceride,
respectively. For each of these esters the same basic reaction is involved, the
reaction of a hydroxyl group with ethylene oxide. This reaction is illustrated in
Equation 8 for a monoglyceride.
A basic catalyst such as potassium hydroxide is used during the ethoxylation process (Egan and Lampost, 1969). The ethoxylation process is a highly
exothermic reaction and adequate cooling must be available to control the reaction temperature. The ethoxylated esters are highly complex in composition,
and the actual components of the ethoxylated sorbitan or monoglyceride esters
have not been defined. Ethylene oxide itself is a highly explosive compound
under certain conditions. Safety instructions on the handling of ethylene oxide
must be read and understood before an ethoxylation process is undertaken.
Another name for ethoxylated compositions is polyoxyethylene (20) sorbitan

CH 2-0H

I \
CH -CH

CH-QH

CH 2-0-CH 2-CHpH

catalyst

II

CH 2-0-C-R

Monoglyceride

1\

CH 2-CH 2

CH-OH

CH -C-R
2 II
0
Ethylene oxide

CH 2-0-CH 2-CHpH

IH2-0-CH2-CH2 -O-CH2-CH2-0H

CH-O-CH 2-CH 2-0H

CH-Q-CH -CH -OH

CH -O-C-R
2

II

(8)

Synthesis and Composition of Food-Grade Emulsifiers

29

monostearate, for example. Polyoxyethylene refers to the ethylene oxide polymeric chain and the (20) indicates that approximately 20 moles of ethylene oxide
have been reacted with one average molecular weight of the starting fatty acid
ester. Other more common names for these products are polysorbate 60 for
ethoxylated sorbitan monostearate, polysorbate 80 for the oleate, polysorbate 65
for the tristearate, and polyglycerate 60 for the saturated monoglyceride.
The polyoxyethylene group is a hydrophilic, or water-loving, group. By the
introduction of the polyoxyethylene group, the starting fatty acid ester is made
more hydrophilic and the final composition has excellent water solubility or
dispersibility. The melting point of each of the starting esters is decreased and
the ethoxylated esters are liquids or soft pastes at room temperature.
The use of the ethoxylated esters in food products is highly regulated, and
the reader is referred to 21CFR172.834, 21CFR172.836, 21CFR172.832, and

21CFR172.840 for details of where and how much polyglycerate 60, polysorbate 60, polysorbate 65, and polysorbate 80, respectively, may be used.

2.8

Succinylated Esters

Succinylated esters of monoglycerides and propylene glycolesters are permitted as direct food additives. The essential step in the preparation of succinylated esters is the reaction of a hydroxyl group with succinic anhydride. This is
illustrated in Equation 9 for a monoglyceride.

II
CH,--c~

Monoglyceride

II

CH 2-0-C-CH 2-CH 2-COOH

+
CH -0-C-R

0 -------+ CH-OH

CH--c/
2

II

Succinic anhydride

CH -0-C-R
2

II

Succinylated monoglyceride

(9)

30

Food Emulsifiers and Their Applications

Freund (1966) patented the manufacture of succinyl monoglycerides; in


this patent he also describes the production of succinyl propylene glycol monoesters. Martin (1968) has patented stearoyl and behenoyl propylene glycol
hydrogen succinate. The succinylation reaction is carried out under anhydrous
conditions with or without a catalyst such as potassium carbonate by the reaction of a distilled monoglyceride or distilled propylene glycol monoester with
succinic anhydride above the melting point of the fatty acid ester and below
the melting point of succinic anhydride. Care must be exercised when handling succinic anhydride because it is an irritant and is sensitive to moisture.
Generally, a reaction temperature of around ll0C is used. If a catalyst is
used, it is generally not removed and is left in the final composition. Care must
be taken to avoid the reaction of the carboxylic acid group of the succinyl
group with other available hydroxyl groups to form polymeric materials. This
polymeric condensation reaction will take place easier in the presence of a
catalyst and at temperatures above 170C. In the determination of the extent of
completion of the succinylation reaction, a nonalcoholic solvent should be
used for an acid number determination. For, if a solvent such as ethyl alcohol
is used, then the alcohol will react with the succinic anhydride, in a fashion
similar to the monoglyceride, and a false indication of the completeness of reaction will be obtained.
While it is possible to react 2 moles of succinic anhydride with 1 mole of
monoglyceride, the most functional results have been found with between
about 0.75 to 1.1 moles of succinic anhydride per mole of distilled monoester,
either monoglyceride or propylene glycol monoester.
The current FDA regulations for succinylated monoglycerides permit any
edible oil or edible fatty acid to be used as a source of the monoglyceride intermediate. However, restrictions are placed on the succinic acid content, acid
number, and melting point of the final composition. Restrictions have been
placed on the use of succinyl monoglycerides in food products; see
21CFR172.830. In contrast, only saturated edible oils predominantly C16 and
C18 in chain length may be used in the preparation of the propylene glycol
monoester intermediate. Regulation 21 CFR172. 765 specifies the acid number,
hydroxyl number, and degree of succinylation of the succistearin (stearoyl
propylene glycol hydrogen succinate). The emulsifier can be used in many applications with good manufacturing practice.

Synthesis and Composition of Food-Grade Emulsifiers

2.9

31

Fruit Acid Esters

There are two fruit acids used to make food-grade emulsifiers: tartaric acid and
citric acid. Tartaric acid-derived esters have been used extensively in Europe,
but they have not been used very much in the United States. One of the foremost uses of tartaric acid-derived esters is a dough conditioner; in the United
States other dough-conditioning agents are used in place of tartaric acid esters.
The tartaric acid esters used are the mono- and diglycerides of diacetyl tartaric
acid (DATEM esters). The synthesis of these esters is shown in Equation 10.

COOH

HO-CH

HO-CH

COOH

Tartaric acid

Acetic anhydride

Diacetyl tartaric
anhydride

Acetic acid

(lOa)

32

Food Emulsifiers and Their Applications

CH -0-C-R
2

0
COOH

H3 C-c-O-CH

H3

I
c-c-o-cz o
""IIc-o CH
2

I
CH-QH

DATEM

(lOb)

The DATEM esters have FDA GRAS status under 21CFR184.ll0l.


However, the regulation states that the DATEM esters must meet the specifications of the Food Chemicals Codex (1981). The Food Chemical Codex specifies the tartaric acid content, the acetic acid content, the total fatty acid
content, the glycerine content, the acid value, and the saponification value of
the DATEM ester. Thus, while DATEM esters have a GRAS status, their composition is limited in the United States.
The acid group of the DATEM esters makes them somewhat sensitive to

Synthesis and Composition of Food-Grade Emulsifiers

33

pH; if the food product is basic, the DATEM ester can be converted to the anionic form. The two acetyl groups are somewhat liable to hydrolysis, which
yields acetic acid. These esters should not be used in aqueous systems that are
expected to have a long storage life.
Two different types of citric acid-based emulsifiers are permitted as direct
food additives: stearyl monoglyceride citrate (21CFR172.755) and monoglyceride citrate (21CFR172.832). The monoglyceride citrate is restricted to use in
antioxidant solutions. It is prepared by the reaction of glycerol monooleate
with citric acid. The final product has an acid number of 70 to 100 and has a
14 to 17% total citric acid content.
Stearyl monoglyceride citrate is prepared by the reaction of citric acid,
monoglycerides, and stearyl alcohol to yield a composition with an acid number of 40 to 52, a saponification number of 215 to 255, and a total citric acid
content of 15 to 18%. The expected reaction is shown in Equation 11.
As can be seen, the reaction products are quite complex. Both saturated and
unsaturated monoglycerides may be used. This type of ester is used in shortenings containing other emulsifiers and is not used extensively in the food industry.

2.10

Acetylated Monoglycerides

Another food-grade additive that is formed by the reaction of an acid anhydride with a hydroxyl group is the acetylated monoglycerides. This reaction is
illustrated in Equation 12.
The starting monoglyceride may be saturated or unsaturated and is of molecularly distilled type. 21CFR172.828 permits the use of a non-food-grade catalyst or a food-grade catalyst in the acetylation reaction. The acetic acid
byproduct and any excess acetic anhydride or formed triacetin are removed by
vacuum distillation. Since the acetic anhydride is monofunctional, there is no
danger of polymers being formed.
The acetylated monoglycerides may also be formed by an interesterification
process. In this case, triacetin is interesterified with a suitable mixture of edible fats or oils and glycerol, as illustrated in Equation 13.
In this case, the composition of the final mixture can be calculated using a random distribution (Feuge and Bailey, 1946). The final reaction mixture is separated by means of steam-stripping and molecular distillation. Any unreacted
triacetin is removed by the vacuum steam-stripping process, and the desired
acetylated monoglycerides are concentrated by the molecular distillation process.

34

Food Emulsifiers and Their Applications

CH 2-COOH

HQ-C-COOH

CH 2-COOH

Citric acid

CH-QH

CH-QH

II

CH-0-C-R

Monoglyceride

Stearyl alcohol

(ll)

Note that in direct reaction acetylation, the hydrophilic hydroxyl group is replaced by a lipophilic acetyl group; thus, if a fully acetylated ester is prepared,
the product is no longer an emulsifier since it has no hydrophilic groups. In the
interesterification process both fully esterified and partially esterified compositions can also be formed, depending on the amount of glycerine used.
The more fully esterified compositions are useful for their film-forming and
moisture barrier properties.
FDA regulation 21CFR172.828 specifies that the final composition have a
Reichert-Meiss! value of 75 to 200 and an acid value under 6. The Reichert-

Synthesis and Composition of Food-Grade Emulsifiers

------+

CH 3-c-o-c-c-CH 3

CH -0-C-R
2

35

+
CH -0-C-R

Monoglyceride

Acetic anhydride

(12)

0
CH 2-0H

I
CH-OH

CH 2-0-C-cH 3

CH 2-0-c-R

CH-O-C-cH 3

I
CH 2-0H

CH-Q-C-R

CH 2-0-C-cH 3

CH-O-C-CH 3

CH 2-0-C-CH 3

catalyst

CH 2-Q-C-R

CH 2-0-C-cH 3

CH-0-C-R
0

II

CH 2-0-C-R

CH-OH

I
CH -0-C-CH

other
compounds

(13)

36

Food Emulsifiers and Their Applications

Meissl number determines the amount of short-chain fatty acids in the final
composition, and this is a measure of the degree of acetylation (see Chapter 3
for methodology). The degree of acetylation in commercially available acetylated monoglycerides ranges from 50 to 90%.

2.11

Phosphated Esters

Information on the synthesis of phosphate esters is sparse. However, Harris


(1939a, b) suggests that they can be prepared by the reaction of a mono- and
diglyceride intermediate with phosphorus pentoxide at 50 to 125C in a twostep reaction. The reaction mixture is then neutralized with sodium carbonate
to produce the desired product.
The monosodium phosphate derivative of mono- and diglycerides of edible
fats or oils or edible fat-forming fatty acids is recognized as GRAS by the FDA.
These emulsifiers are available in a solid, liquid, or paste form. Typically, the
phosphorus content of these compositions ranges from 3.5 to 5.5%, and their pH
ranges from 6.0 to 7.5. They have found some applications in the confectionery
industry as viscosity reducers analogous to lecithin (Chapter 9).

2.12

Sucrose Esters

The newest family of emulsifiers to obtain FDA approval for use as direct food
additives is the sucrose esters. This occurred in 1982 (47FR5547). The sucrose esters permitted as additives under 21CFR172.859 are the mono-, di-,
and triesters. This regulation does not relate to the fully esterified sucrose esters that have been promoted as nonabsorbable fat and oil replacements. These
esters are a mixture of the octa-, hepta-, and hexaesters of sucrose.
The main method of preparing sucrose monoesters is by means of an interesterification process between sucrose and methyl esters of fatty acids. The basic process is illustrated in Equation 14.
Sucrose and the methyl esters of fatty acids are very insoluble in one another. In addition, sucrose darkens and carmelizes on heating to temperatures
around 150C. This insolubility and darkening of the sucrose has led to the
use of organic solvents such as N,N-dimethylformamide (DMF) or dimethyl
sulfoxide (DMSO) and reduced pressure in the preparation of the sucrose
mono-, di-, and triesters (Haas, 1959). The use of DMSO as a solvent was allowed in 1987 (52FR10883).

Synthesis and Composition of Food-Grade Emulsifiers

37

HOCH 2

HO
H

OH

OH

Sucrose

II

R-C-OCH 3 _______.

Methyl esters

RCOOCH 2

HO
H

Sucrose monoester

OH

OH

Methyl alcohol

(14)

Another interesterification process used to prepare the sucrose monoesters


under vacuum is known as the "microemulsion process," or the "transparent
emulsion process." In this process edible propylene glycol is utilized as a solvent and reactant (Osipow and Rosenblatt, 1969). In an improvement on the
original microemulsion process, it was found that water could be used under
carefully controlled reaction conditions in place of the propylene glycol (Osipow
and Rosenblatt, 1972; Yamagishi et al., 1974). In the microemulsion process, a
very high amount of a sodium or potassium salt of a carboxylic acid is used to

38

Food Emulsifiers and Their Applications

produce an emulsion particle size less than one-quarter the wavelength of light,
in contrast to other emulsions in which particle sizes are greater than the wavelength of light. This particle is so small the emulsion appears transparent.
However, the use of the high amount of the metal salts of the fatty acids and unreacted sucrose leads to difficult purification and separation procedures. In general, solvents such as ethyl acetate, methyl ethyl ketone, or isobutyl alcohol are
used during the purification process. FDA rule 21CFII 72.859 specifies the
amount of these solvents that can be in the final sucrose ester. A patent has been
issued regarding the purification of the sucrose ester reaction mixture (Mizutani
et al., 1973), but many other purification processes are possible. Overall, the
production of the sucrose monoesters is a more complicated process than the
production of the other food-grade emulsifiers.

References
Babayan, V.K., Lehman, H. (1973). U.S. Patent 3,637,774, January 25.
Barsky, G. (1950). U.S. Patent 2,509,414, May 30.
Brown, K.B. (1943). U.S. Patent, 2,322,820, June 29.
Egan, R.R., Lampost, S.B. (1969). U.S. Patent, 3,433,645, March 18.
Eng, S. (1972). U.S. Patent 3,636,017, January 18.
Federal Register, 53, 40808, December, 1988.
Feuge, R.O., Bailey, A.E. (1946). Oil and Soap, 23, 259.
Food Chemical Codex (1981). 3d ed., pp. 98-99.
Freund, E.H. (1966). U.S. Patent 3,293,272, December 20.
Harris, B.R. (1939a). U.S. Patent 2,177,983, October 31.
- - (1939b). U.S. Patent 2,177,984, October 31.
- - (1941). U.S. Patent 2,258,892, October 14.
Hass, H.B. (1959). U.S. Patent 2,893,990, July 7.
Japanese Patent Publication (1974). No. 15246, April13.
Martin, J.B. (1968). U.S. Patent 3,375,262, March 26.
Mizutani, N., et al. (1973). U.S. Patent 3,798,324, July 24.
Osipow, L.l., Rosenblatt, W. (1969). U.S. Patent 3,480,616, November 25.
- - (1972). U.S. Patent 3,644,333, February 22.
Stockburger, G.J. (1981). U.S. Patent 4,297,290, October 21.
Thompson, J.B., Buddemeyer, B.D. (1956). U.S. Patent 2,744,825, May 8.
Thompson, J.B., eta!. (1956). U.S. Patent 2,733,252, January 31.
Yamagishi, F., eta!. (1974). U.S. Patent 3,792,041, February 12.

THREE

Analysis of Food Emulsifiers


Gerard L. Hasenhuettl

3.1

Introduction

Analytical methods for food emulsifiers are closely associated with or derived
from methods commonly used for fats and oils (Sonntag, 1982; Karleskind,
1996). Test methods are of several types and are carried out for a variety of
reasons. Analysis ensures that the composition of the emulsifier is correct and
that it has not seriously degraded during processing. Often the composition
and distribution of homologs has implications for the utility of the emulsifier in
the individual food product (see, for example, Halkier, 1980). The final level of
testing is often a measurement of performance in the food system itself.
Specifications are negotiated and agreed upon between the producer and the
customer (usually a processed food manufacturer). Analytical tests are carried
out on the process line or control laboratory of the supplier, who then provides
a certificate of analysis to the customer. The customer may then check the
analysis as part of the receiving procedure and accept or reject the shipment.
Disputes may be settled by submitting a sample to an independent laboratory.
Quality assurance testing is also necessary to ensure that foods conform to

39

40

Food Emulsifiers and Their Applications

standards set by government regulatory agencies. The U.S. Food and Drug
Administration (FDA), the European Economic Community (EEC), and the
regulatory agencies of individual countries have limits on the maximum concentration of emulsifiers or other food additives allowed in specific products.
This topic has been reviewed by several authors (Kroeller, 1966, 1968;
Gernert, 1968; Amano, 1979). Professional organizations, such as the
Association of Official Analytical Chemists (AOAC), the American Oil
Chemists' Society (AOCS), International Union of Pure & Applied Chemistry
(IUPAC), Leatherhead Food Research Association, and the National Academy
of Sciences (Food Chemicals Codex), have compiled lists of official methods
that are used for fats, oils, and emulsifiers (Cunniff, 1995; Firestone, 1990;
Paquot and Hauffen, 1987; Slack, 1987; Taylor, 1996).
To determine emulsifiers in intact food products, the fats and emulsifiers must
first be extracted. Fats and oils are soluble in nonpolar solvents, such as hexane
or toluene. However, emulsifiers are amphiphilic and more polar and tend to
be less soluble in cold hexane or petroleum ether, particularly when emulsifier
concentration is high compared to total lipid level. Chloroform or chlorofonn/
methanol mixtures (Flor and Prager, 1980) are sufficiently polar but also sufficiently immiscible to allow for extraction from aqueous food systems. In cases
where total lipid content is high compared to emulsifier concentration,
Halverson and Qvist (1974) have reported extracting total lipids with refluxing
hexane followed by extraction with acetonitrile. Solid foods samples (e.g., cake,
coffee whitener) are conveniently extracted using a Soxhlet extraction apparatus.
Liquid samples (e.g., milk, ice cream mix) are usually separated in a separatory
funnel or countercurrent distribution apparatus. Another factor complicating extraction from foods is that lipids may be tightly complexed by starches or proteins or may simply be encapsulated in a biopolymeric matrix. Jodlbauer (1976)
has demonstrated the use of an amylase enzyme to release lipids from a pasta
product so that they are more easily extracted.

3.2 Thin-Layer and Column Chromatography


After lipids have been extracted, the emulsifiers may be isolated from the nonpolar lipids by simple thin-layer or column chromatography. For example, on a
silica gel column, triglycerides may be eluted with hexane followed by diglycerides with 5% diethyl ether, and monoglycerides in 10% ether/hexane.
Dieffenbacher et al. (1988, 1989) have described a one-step extraction and

Analysis of Food Emulsifiers

41

purification process using a Celite/sodium sulfate column. Fractions from


these separations may be quantitated gravimetrically and/or subjected to further analytical testing.
Thin-layer chromatography (TLC) and paper chromatography have been
used for detection of food emulsifiers. The method was a simple, rapid, and
reasonably reliable means to test for the presence of emulsifiers in lipid components (Murphy and Hibbert, 1969; Wurziger, 1968). Mono- and diglycerides
are readily separable on a boric acid-impregnated silica gel plate. Spots can
be visualized by spraying with dichlorofluorescein and identified by their Rf
values. Schmid and Otteneder (1976) utilized a petroleum ether, diethyl ether,
and acetic acid solvent system to identify monoglycerides in alimentary pastes.
Kanematsu and coworkers (1972) separated monoglycerides from propylene
glycol fatty acid esters in shortenings using a chloroform/acetone solvent system. Thin-layer chromatography was also used by Paganuzzi (1987) to separate
lipids for further analysis by gas-liquid chromatography.
Because of their importance in lipid metabolism and as components of lipid
membranes, phospholipids have been widely studied and analyzed. TLC has
been reported as a method of separation and analysis of phospholipids from
fish and oilseeds (Erdahl et al., 1973; Vyncke and Lagrou, 1973; Kimura et al.,
1969; El-Sebaiy et al., 1980; Lendrath et al., 1990; Biacs et al., 1978).
Watanabe and coworkers (1986) have reported a two-dimensional TLC procedure on silica gel that uses acidic and basic solvent systems. The method was
applied to soy and egg lecithins. Detection of phospholipids may be accomplished by conventional nonselective stains, such as sulfuric acid. However,
the compound class may be distinguished from other lipids by phosphorus detection, by either selective reagents or spectroscopy (Senelt et al., 1986).
Experimental design can also be used to optimize separation of phosphoruscontaining lipids (Olsson et al., 1990).
Monoglycerides can be further derivatized with organic acids to produce
new emulsifiers with unique functionalities (see Chapter 2 for synthesis).
Compounds such as lactylated, succinylated, and diacetyltartaric esters
(DATEM) monoglycerides may be determined by TLC (Bruemmer, 1971;
Yusupoca et al., 1976; Jodlbauer, 1981). These methods deal with the complexity of separating derivatives from unreacted monoglycerides. In some
cases, synthesized authentic standards were used to confirm identification of
the spots. Calcium and sodium stearoyllactylates, also widely used in bakery
products, are also capable of being determined by TLC. Regula (1975) carried

42

Food Emulsifiers and Their Applications

out the separation on Kieselgel G using a hexane/acetone/acetic acid solvent


system. Spots were visualized by spraying with bromocresol green. Kroeller
(1969) used starch paper impregnated with mineral oil as the stationary phase.
Methyl red was used to visualize the top half of the chromatogram, and potassium hexacyanoferrate was used on the bottom.
One of the major drawbacks to the routine use of TLC in lipid analysis has
been the difficulty of obtaining quantitative rather than qualitative results.
This problem has been addressed by the development of a unique chromatographic system that uses a coated rod rather than a plate. After development,
the rod is placed in a scanning-flame ionization detector that transforms spots
into peaks on an x-y plot. Determinations of monoglycerides (Takagi and
ltabashi, 1986; Ranny et al., 1983), phospholipids (Tanaka et al., 1979), and
sucrose esters (Rios et al., 1994) have been reported using this technique.
Duden and Fricker (1977) have described the detection of monoglycerides and
lecithin using a fluorescence reagent in a chromatogram spectrophotometer.
Despite these improvements, TLC is being rapidly replaced by gas-liquid
chromatography (GLC) and high-performance liquid chromatography. These
methods offer the convenience of autosampling, which translates into improved productivity for routine analyses.
Historically, the first methods developed for testing fats, oils, and emulsifiers were wet methods; that is, they involved the use of solutions and chemical
reactions. Procedures used to define these tests often involved separations and
structure determination of the products through chemical degradation. Tests to
measure the melting properties of these materials were spawned in the early
1900s as an adjunct technology of catalytic hydrogenation. With the development of gas-liquid chromatography and high-performance liquid chromatography, instrumental methods have often replaced older, more labor intensive wet
methods. Automated sampling served to increase analytical productivity even
further. Perhaps the ideal model of a control lab is a clinical chemistry facility.
Automated sampling, fluid transfer, reactions, determination, and reporting of
results have virtually eliminated repetitive procedures.
The major problem with control testing has been that it occurs after the
emulsifier has been produced. The results then compel the quality control
manager to accept or reject the product. Modem quality practices are incorporating analytical measurements directly into the process stream or chemical
reactor. In-process analysis allows rapid correction before the product is produced, resulting in less scrap and rework.

Analysis of Food Emulsifiers

3.3

43

Wet Chemical Analysis

Wet analysis describes a determination that uses solvents and chemical reagents
to determine an aspect of emulsifier composition. An example would be titration
of free fatty acid with alcoholic potassium hydroxide. Wet methods have been
standard practice for many years. However, a large analysis volume in laboratories poses the problem of disposal or recycling of reagents and/or spent solvents.
For this reason, the current trend is to replace the wet method with instrumental
or physical measurements that supply related or equivalent information. Some of
the most widely used methods for emulsifiers are described next.

3.3.1

a- Monoglyceride t

Monoglycerides may exist as alpha, the most predominant form, which has the
fatty acid esterified in the 1 or 3 positions of the glycerol head group, or beta,
where the substitution is at the 2 position. The alpha isomer has adjacent hydroxyl groups in the 1, 2 or 2, 3 positions. The isomer is determined by titration
with periodic acid resulting in cleavage, as shown in the following equation:
The analysis is carried out by dissolving the sample (extracted lipid or pure

OH

I
OH

0 -1-1 C17 H35

----+

HI0 4

II

-c-c

CH

0 -~-~C17 H35

HI04

HI03

Hp

(15)
t

Cunniff (l995a); Firestone (l990n).

44

Food Emulsifiers and Their Applications

emulsifier) in a solvent and reacting with an excess of periodic acid dissolved


in 95% methanol. Potassium iodide is then added, and liberated iodine is
titrated with standard arsenite solution. An aqueous solution of periodic acid
is used to determine free glycerine. The result for monoglyceride is then corrected for the amount of periodic acid consumed by free glycerine. For many
years, chloroform, a potential carcinogen, was the solvent used in this test.
Since about 90% of the monoglyceride is alpha, this determination is an accepted approximation of the total monoglyceride concentration. Due to the
problem of solvent disposal and the fact that the method is labor intensive, it
has been largely supplanted by gas-liquid chromatography (Firestone, l990g)
(see Section 3.5, Instrumental Methods). AOCS has classified the test as a surplus (obsolete) method.

3.3.2

Acid Value/Free Fatty Acid t

Fatty acids may occur during the preparation of many common food emulsifiers. They may be present as residues from direct esterification reactions or
may be due to hydrolysis of glycerides. Fatty acids may act as antiemulsifiers
in some systems. They also make a system sensitive to pH, since alkali transforms fatty acids into soaps, which are powerful anionic surfactants. Attempts
are therefore made to keep free fatty acids in mono/diglycerides as low as possible. Other emulsifiers, such as sodium stearoyl lactylate and lactylated
monoglycerides have high acid values that must fall in a range defined by the
FDA or other regulatory body.
Acid value is determined by dissolving a weighed sample of the emulsifier
in an isopropanol/toluene or ethanol/diethyl ether solvent and titrating with a
standard potassium hydroxide solution to a phenolphthalein endpoint. In cases
where the acid value is used to track the progress of reactions using anhydrides, such as in production of DATEM and acetylated monoglycerides, an
aprotic solvent rather than an alcohol must be used. Alternatively, the sample
solution may be titrated potentiometrically to the equivalence inflection point.
The latter method is particularly useful for dark-colored samples where a visual endpoint is difficult to observe. The acid value is defined as the number of
milligrams of potassium hydroxide necessary to neutralize the free fatty acid
per gram of sample and is determined by the formula

+Cunniff (l995c); Firestone (1990k).

Analysis of Food Emulsifiers

45

(A- B)(N)(56.1)

w
where A

mL standard alkali used in titration

B = mL standard alkali used to titrate blank

N = normality of standard alkali


W = g of sample
Free fatty acid percentage can be determined by dividing the acid value by
a factor characteristic of the fatty acid present. Values for this factor are C12
(lauric)= 2.81, C16 (palmitic)= 2.19, and C18 (stearic or oleic)= 1.99.

3.3.3

Iodine Valuet

Emulsifiers may be prepared from saturated or unsaturated (mainly oleic) acid


or from partially hydrogenated triglycerides. Unsaturated emulsifiers are more
susceptible to oxidative rancidity than those derived from saturated fats or
fatty acids. Titration with reagents that add across the carbon-carbon double
bond have been utilized to determine unsaturation since the early days of organic chemistry. Two common methods have gained common acceptance. (i)
The Wijs method reacts the sample dissolved in carbon tetrachloride with an
iodine monochloride solution in glacial acetic acid; excess reagent is then
titrated with standard sodium thiosulfate solution. (ii) The Hanus method is almost identical but uses iodine monobromide as the reagent. Because of the
high toxicity of carbon tetrachloride, a modified method has been developed
that substitutes cyclohexane. Iodine value is expressed as the number of centigrams of iodine absorbed per gram of sample (same as % iodine absorbed) and
is calculated using the formula

(S- B)(N)(l2.69)

where

s=

w
mL used to titrate sample

B = mL used to titrate blank


N = normality of thiosulfate solution

w=
t

weight of sample

Cunniff (1990e); Firestone (l990f).

46

Food Emulsifiers and Their Applications

In this test, when reporting the value, it is important to also identify the
method by which the value was determined.
Iodine value may also be calculated from the fatty acid composition (FAC) of
fats, oils, and fatty acids through gas-liquid chromatography (GLC)(Firestone,
1990h). The same method may be used for food emulsifiers, but new conversion
factors would probably need to be developed to translate GLC peaks to iodine
values. Near infrared reflectance (NIR) has also been developed as a rapid
method to determine unsaturation by measurement of the carbon-carbon double
bond absorbance peak. However, a great deal of work is necessary up front to assemble a calibration curve to relate the measured absorbances to traditionally
measured values.

3.3.4

Peroxide Valuet

As mentioned before, unsaturated fatty acids can undergo oxidation to produce


off-flavors characteristic of rancidity. Autoxidation occurs through a free radical
chain reaction that produces hydroperoxides as an early intermediate. The hydroperoxide can react further to produce aldehydes, ketones, and hydrocarbons.
Detection of hydroperoxides in food ingredients is therefore a necessary precaution to ensure that the product does not become rancid during its normal shelf life.

Peroxides and hydroperoxides may be determined by the reaction of a solution of the sample in acetic acid and chloroform with potassium iodide.
Liberated iodine is then titrated with standardized sodium thiosulfate solution
using a starch indicator. Precautions need to be taken to avoid the presence of
residual oxidizing or reducing agents on glassware. Also, strong ultraviolet
light must be avoided, since it forms peroxides by a photochemically induced
oxygen addition. Peroxide is defined as the number of milliequivalents of peroxide (AOAC uses the term "active oxygen") per 1000 g of sample that oxidize
potassium iodide. The exact value is obtained from the following formula:

(V)(T) (1000)
m

where V = volume of titration


T = normality of thiosulfate solution

m =

wt

in g of sample

Cunniff (l995g); Firestone (1990s).

Analysis of Food Emulsifiers

47

Since the practicing formulator needs to minimize the peroxide content of ingredients, the lower the peroxide value, the less is the risk of developing rancidity.
Special surveillance should be maintained on materials that have higher iodine
values (higher unsaturation). Of course, other materials that catalyze oxidation,
such as iron, copper, and lipoxygenase enzymes, must also be considered.
Recently, peroxides have been determined utilizing high-performance liquid chromatography (HPLC) (Yang, 1992; Yang et al., 1991). A sample is separated on a column and fractions then pass through a post-column reaction
chamber where they are mixed with a luminescence reagent. Light emitted
from the chemiluminescence reactions is then measured by a special photochemical detector. The method is fairly rapid and requires only small amounts
of sample. An additional advantage is that different peroxide species may be
measured separately, which may indicate where oxidation is occurring. The
major disadvantage is the capital cost of the equipment and the high cost of the
luminescence reagent.

3.3.5

Saponification Numbert

Reaction of a fat, oil, or emulsifier with hot alcoholic potassium hydroxide


causes the cleavage (saponification) of fatty acids from the polyol portion of the
lipid. The extent of this cleavage can be measured by back titration with dilute
hydrochloric acid with a phenolphthalein indicator. Alternatively, the titration
can be potentiometric, and the endpoint determined by observation of the inflection point. This procedure is especially convenient for dark samples where
the phenolphthalein endpoint is difficult to see. Saponification value is the
number of milligrams of potassium hydroxide required to saponify one gram of
fat and is determined by the formula

(B- S)(N) (56.1)

where B = mL used to titrate blank


= mL used to titrate sample

N = normality of standard alkali

w=
t

wt of sampling

Cunniff (1995d); Firestone (1990j).

48

Food Emulsifiers and Their Applications

This value is sensitive to both the degree of esterification and the chain
length of the substituent fatty acid. Higher degrees of esterification and shorter
chain lengths are reflected by higher saponification values. Conversely, longer
chain lengths and lower degrees of substitution are reflected in lower saponification values.

3.3.6

Hydroxyl Valuet

Free hydroxyl groups may be determined by reaction of the emulsifier with


acetic anhydride in the presence of pyridine. Water is then added and heated
to hydrolyze excess acetic anhydride to acetic acid (as well as mixed anhydrides to acetic and fatty acids). The mixture is then titrated with standard alkali to determine residual acetic (and fatty) acid. The hydroxyl value is
determined by the formula

_(5_6._1)_(7)_(V._
0 -_V_) + AV
m
where T = normality of standard alkali solution

V0 = mL of standard alkali to titrate blank


V = mL of standard alkali to titrate sample
w = wt of sampling

AV = acid value of sample

It is recommended that the value be obtained by averaging duplicates. In a


sense, the hydroxyl value provides inverse information to the saponification
value. High saponification value indicates a high degree of substitution, while
high hydroxyl values are indicative of low degrees of substitution.
Major difficulties with the hydroxyl method are (i) a great deal of operator
skill and experience is required, (ii) the method is very time-consuming and
labor intensive, and (iii) results are often inconsistent due to the differential
reactivity of primary and secondary hydroxyl groups. A near infrared reflectance (NIR) method that uses the hydroxyl-stretching frequency is an attractive alternative. It requires only 5 to 10 minutes to obtain a value and is
therefore more adaptable to process control than the titrimetric method. The
only drawback is the necessity of constructing a calibration curve when setting

Cunniff (1995!); Firestone (19901).

Analysis of Food Emulsifiers

49

up the determination. Hydroxyl values are used mainly to specify and control
polyglycerols and polyglycerol esters.

3.3.7

Lactic Acid Analysest

Monoglycerides and fatty acids may be reacted with lactic acid to produce
lactylated derivatives that are useful emulsifiers. Lactic acid may be present as
esterified or free lactic acid. In addition, lactic acid has both a carboxyl function and can self-condense to produce small polymers. The total lactic acid
present in a sample encompasses all these species and is indicative of the lactic acid/lipid ratio in the production batch. In this test, a sample is boiled with
alcoholic potassium hydroxide followed by addition of hydrochloric acid solution. Lipid material is separated by exhaustive extraction with diethyl ether.
The aqueous phase is then titrated with standard potassium hydroxide solution
to the phenolphthalein endpoint.
Free lactic acid is determined by dissolving the sample in benzene and extracting the solution with water. Free lactic and poly lactic acids are then quantitated by titration with standard potassium hydroxide. An obvious problem
with this test is the use of benzene, which is a carcinogen.
Water-insoluble combined lactic acid (WICLA) is determined by dissolving a
sample in benzene and extracting the sample as described in the previous test. The
benzene layer (or upper phase from the previous test) is evaporated to dryness. The
residual solid is then heated with alcoholic potassium hydroxide neutralized and
titrated in the same manner as carried out in determination of total lactic acid.
The above methods suffer from the disadvantage of using a toxic, carcinogenic solvent and laborious phase-separation procedures. Franzke and Kroll
(1977) recommend the use of chloroform and petroleum ether for the determination of free and total hydroxy acids, respectively. The authors report the
analysis of lactic, citric, and tartaric acid in emulsifiers. They have also reported the use of an enzymatic method for determination of total lactic acid
(Franzke and Kroll, 1980).

3.3.8

Reichert-Meiss! Valuet

This method is specific for determination of acetic and short-chain fatty acids that
have been esterified to mono- or diglycerides. The method involves (i) cleavage of

Taylor (l996a).

50

Food Emulsifiers and Their Applications

the short-chain acid in alkali followed by neutralization with dilute sulfuric acid,
(ii) distillation of the liberated acetic (or other short-chain) acid, and (iii) determination of the condensed acid by titration with standard base to the phenolphthalein
endpoint. The method is therefore equipment intensive (the condensing unit must
be replaced or cleaned between samples) and time-consuming.

3.3.9

Moisture

Moisture is generally undesirable in food emulsifiers because it is a diluent and


it represents a potential problem for microbial growth. Some exceptions are the
polysorbates that require a small amount of water to provide clarity and prevent
phase separation, and emulsifier hydrates where gel phases are preformed for
specific applications. Several methods for moisture have been developed through
the history of oil processing. The distillation method uses a toluene solvent to assist distillation of water from a large sample through a condenser and into a graduated separation tube (Firestone, l990i). The water is quantitated by volume.
The method is equipment intensive because of the need to clean and replace the
distillation apparatus between samples. The distillation method is most
amenable where the water concentration is about 0.5% or higher.
Lower levels of moisture are readily analyzed by reaction with Karl Fischer
reagent. The procedure is amenable to lecithin as well as other emulsifiers that
can be solubilized for the analysis (Firestone, l990p). Instruments are currently available that allow repetitive determinations without the necessity of
disassembly and cleaning between samples.

3.3.10

Color

Color of emulsifier is a reflection of the quality of starting lipid in its manufacture and/or manufacturing conditions. Thermal abuse of fats and emulsifiers
are well known to produce dark colors. In many cases, oxidized tocopherols
are thought to be the chromatic impurities. Dark-colored emulsifiers may detract from the appearance of finished food products where they are used in
high concentrations (e.g., cake icings). However, color evidence that an emulsifier has been abused also suggests that pro-oxidants may be present that may
adversely affect the flavor of the product. Emulsifiers would therefore be
processed to produce the lightest color possible.

Firestone (l990m).

Analysis of Food Emulsifiers

51

Color is most commonly determined by comparison with a set of standards,


such as colored glass. The most common example is the Lovibond method (also
known as the Wesson method) in which a column ofliquid (or molten) emulsifier
is compared to a set of colored standards (Firestone, l990b). Two values are obtained for red (R) and yellow (Y) colors. These are the most common colors encountered in thermally abused emulsifiers. A related method, which reports only
one value, is the Gardner method (Firestone, l990t). The method is most commonly used for measurement of color in lecithin, but otherwise is widely applied
to nonedible products. Spectrophotometric methods, such as ultraviolet absorption, have not been widely accepted, probably because the information they provide is ambiguous. Many constituents commonly found in lipids can absorb in
the ultraviolet region. Spectrophotometry is useful in determination of green pigments such as chlorophyll (Cunniff, l995b; Firestone, l990c). Since spectrophotometric methods are most amenable to adaptation for on-line process control, it
is likely that their use will increase as processing becomes more advanced.

3.3.11

Fatty Acid Soaps

Although the official AOCS method has been classified as a surplus method
(obsolete), residual soap remains an important process check in emulsifier production. Many emulsifiers are produced by interesterification using alkaline
catalysts (see Chapter 2). The catalyst is neutralized at the end of the process
prior to removal of excess polyol by distillation. In batch processes, if the alkali is not properly neutralized, disproportionation (reverse reactions) can occur, resulting in emulsifiers that are out of specification. Analysis of soap
(alkali) is used to assess proper neutralization.
The determination is carried out by dissolving an emulsifier sample in a
suitable organic solvent/water mixture and titrating with standard hydrochloric
acid. Bromophenol blue or phenolphthalein may be used as indicators. The
method may also be adapted for potentiometric titration. Another method for
assessing the extent of neutralization is to measure the pH. A 5% solution of
emulsifier in isopropanol/water is equilibrated to room temperature and the pH
is read using a standard electrode. A pH reading of 6.5 to 6.8 indicates that
soap is absent and the emulsifier has been properly neutralized.

3.3.12

Phosphorus and Other Lecithin Procedures

Soy lecithin, which is widely used as an emulsifier in food products such as


chocolate and margarine, is obtained by degumming of soybean oil. Purity of

52

Food Emulsifiers and Their Applications

the lecithin obtained in this way is therefore a major concern. Its chemical
structure is that of a phospholipid where phosphoric acid is esterified to a glycerol molecule and an organic base. Total phosphorus in the sample may be determined by a laborious process in which the sample is saponified, phosphorus
is separated by precipitation with molybdate solution, the precipitate is dissolved in base, and excess base is titrated with standard nitric acid (Firestone,
1990r). Tamanaka and Kudo (1991) have reported a rapid method where phosphorus precipitated by molybdate is determined spectrophotometrically. A
much simpler, albeit less precise, procedure is to determine lecithin content as
acetone-insoluble material based on the principle that neutral lipids are soluble in acetone while phospholipids are not (Firestone, 1990g). This principle
was used by Goldstein (1984) to develop a method for determination of lecithin
in oils. The oil sample was dispersed in acetone and lecithin was quantitated
by measurement of the turbidity.
Individual

phosphatides,

such

as

phosphatidylcholine

and

phos-

phatidylethanolamine may also be determined by HPLC (see Section 3.5,


Instrumental Methods). Combinational measurements have been reported for
separation and phospholipid class identification in commercial soy lecithin
(Lendrath et al., 1991) and in egg yolk (Holopainen, 1972). Individual phospholipids of soybean and rapeseed were separated by micellar electrokinetic
capillary chromatography (lngvardsen and Michaelsen, 1994).
Determining the presence and composition of phospholipids in foods is an
exercise in separation followed by measurement. Wewala and Baldwin (1982)
describe a procedure for the analysis of lecithin in instant milk powders.
Ugrinovits (1983) described determination oflecithin in cocoa powders.

3.4

Physical Methods

Physical properties of emulsifiers are often critical to their functionality and


subsequent consumer acceptance of finished food products. The next chapter
will explore this topic in some detail. In this section we will discuss analytical
procedures for a few routinely measured properties.

3.4.1

Melting Point

The temperature at which an emulsifier melts may have a substantial impact on a


food-processing operation. For example, the melting point of a peanut butter stabilizer must be matched to the fill temperature of the peanut butter to prevent oil

Analysis of Food Emulsifiers

53

separation. Several methods are commonly used to determine melting points of


fats and emulsifiers. Unlike pure chemical compounds these materials exhibit
broad melting ranges rather than sharp melting points. To further complicate the
situation, phases may melt and then recrystallize as polymorphic forms undergo
transitions in the system. Techniques such as differential scanning calorimetry
and programmed-temperature x-ray diffraction are necessary to study these
polymorphic transitions, but they will not be discussed here.
Capillary melting points, widely used for other substances, are sometimes
utilized. The range is usually observed as an initial softening progressing gradually to complete liquefaction. The melting point is defined as the temperature
at which the sample first becomes a clear liquid (Firestone, l990a).
Shortcomings of this method are that it is subjective, it is susceptible to variations in heating rate, and dispersed inorganic material interferes with observation of the clear liquid. Softening point, also referred to as the slip point, has
been used to determine melting behavior of fats and oils. The procedure is
similar to capillary melting point, except it uses an open capillary, and the
endpoint is the temperature at which the sample runs out of the tube. However,
collaborative studies have found it to be unsuitable for emulsifiers. The congeal point (Firestone, l990d) is determined by cooling a relatively large sample of fat to a first crystallization temperature. A temperature curve is then
constructed as a crystallization exotherm is observed. The maximum temperature of the curve is defined as the "congeal point." This method is less sensitive to insoluble impurities but is time-consuming when performed manually.
Wiley melting point is a long accepted method for emulsifiers as well as for
fats and oils. The measurement involves suspending a disc of material at an interface of unmixed water and ethanol. Heating and gentle stirring with a thermometer are applied. When the disc melts and becomes spherical, the
temperature is read and reported. Definition of the exact temperature where
the solid becomes a sphere is somewhat operator-dependent and the method is
also time-consuming. AOCS has classified the method as surplus (obsolete).
Mettler dropping point also starts with a disc of emulsifier that is placed in
a disc holder. The disc is heated in an instrument at a specified rate. When the
disc melts, it drops out of the holder and breaks an electric eye circuit and the
temperature is recorded. Advantages of this method over other techniques are
(i) it is unaffected by operator variability in determining the endpoint, (ii) the
measuring process can be left unattended, and (iii) output can be recorded into
a computer control chart thus eliminating transcription errors.

54

Food Emulsifiers and Their Applications

3.4.2

Viscosity/Consistency

Viscosity is a property of an emulsifier that is important to processing and transfer


of material, such as pumping through product feed lines. Measurements are not
generally made routinely. A viscosity/temperature profile is usually constructed,
and acceptable ranges are defined for processing operations. Some classes of
emulsifiers, such as polyglycerol esters, are very thick and have to be heated to be
pumped through heat-traced lines. Sucrose stearate poses a problem since it decomposes when heated too high and is therefore preferred as a powder form.
Viscosity of lecithins had been measured by a procedure known as the "bubble time" method (Firestone, l990v) that was also used widely in the paint industry. A clear liquid sample is placed in an ASTM tube in a constant-temperature
bath. The tube is inverted to produce a bubble at the bottom of a column ofliquid.
The time for the bubble to reach the top is measured and recorded. The value in
poise is obtained by comparison with standard samples. Viscosity may also be determined directly using a Brookfield viscometer (Firestone, 1990u). In the latter
case, it is not as critical that the samples be clear.
Rheological properties are most often determined when emulsifiers are incorporated into foods and ingredients such as shortenings. Consistency (Firestone,
l990e) may be measured using a penetrometer, such as an lnstron or Stevens
texture analyzer. The method measures the distance a probe penetrates into a
sample at a given force, temperature, and time. This property is critical to performance when the product is mechanically incorporated into a food product. An
example is dough-sheeting to produce Danish pastries or pizza crust.

3.4.3

Refractive Index

Production of some emulsifiers is controlled by refractive index measurement.


For emulsifiers, in most cases, the measurement is correlated to some other
property. For example, in the preparation of polyglycerol, refractive index increases with the degree of polymerization. The method is rapid and much more
convenient than a wet chemical method such as hydroxyl number. Since many
food emulsifiers are solids, it is convenient to maintain the refractometer at a
temperature of 60C for all measurements.

3.5

Instrumental Methods

There has been little recent effort toward development of new wet methods for
analysis of food emulsifiers. Studies that are carried out in this area have fo-

Analysis of Food Emulsifiers

55

cused almost exclusively on elimination of toxic solvents or substitution with a


more benign solvent. Instrumental methods that provide a rapid unattended
process are attractive areas of investigation. In addition, some of the more sophisticated terchniques can separate lipid classes and even distinct compounds. The disadvantage of instrumental methods is the high capital cost and
maintenance of sophisticated instruments. In this section, we will provide an
overview of a number of instrumental methods for characterizing emulsifiers.

3.5.1

Gas-Liquid Chromatographyt

Volatile compounds may be easily separated and detected using a gas-liquid


chromatograph. However, food emulsifiers and lipids in general are nonvolatile. To usefully apply this analytical technique, emulsifiers must be derivatized to increase their volatility. Structural information may also be
obtained by cleavage of the emulsifier followed by derivatization and analysis
of its subunits.
The most common GLC analysis performed on lipids is the fatty acid composition (FAC). In this test, fatty acids are cleaved from their glycerol or other
polyol backbone and converted to a volatile derivative, most often methyl esters. The sample is injected and separated on a packed or capillary column
and detected on a flame ionization or thermal conductivity detector. Fatty
acids are well separated primarily by chain length and secondarily by unsaturation. Peaks are identified by their retention times as previously established
by a standard. Concentration is determined by peak height or area corrected
by response factor as a percentage of the total fatty acids.
Monoglycerides and propylene glycol esters of fatty acids can be determined directly by derivatization of the dry sample by hexamethyldisilazane
and chlorotrimethylsilane in the presence of pyridine (Nakanishi and Tsuda,
1983). Dieffenbacher has suggested the use of N,N-bis(trimethylsilyl)trifluoroacetamide (BSTFA) in a 3:1 ratio with chlorotrimethylsilane (Brueschweiler
and Dieffenbacher, 1991). Lee and coworkers (1988) have formed propyl derivatives of mono- and diglycerides and determined them by GC/MS. After
separation of the sample, peaks are quantitatively determined by reference to
an internal standard. Monoheptadecanoylglycerol (monomargarine) had been
traditionally used but it is difficult to synthesize. Solutions of this standard are
not stable and must be periodically prepared. Butyl alcohol, which has the

Christie (1989).

56

Food Emulsifiers and Their Applications

same molecular weight as monomargarine, is commercially available at a reasonable cost and stable to 1,2-acyl shifts (Hasenhuettl, 1990). Cholesteryl acetate has also been utilized as an internal standard (Blum and Koehler, 1970).
Collaborative studies have been conducted with the objective of developing a
uniform method for AOCS, AOAC, and IUPAC (Brueschweiler and
Hautefenne, 1990; Firestone, 1994). Figure 3.1 shows the separation of propylene glycol esters and monoglycerides by GLC.
Polyol distributions of other food emulsifiers can be determined by cleavage of fatty acids, as for fatty acid distribution. The residual polyol can be purified and converted to a volatile trimethylsilyl derivative that is analyzed by
GLC. Use of this technique provides structure information on the underlying
polyol. Percentages of sorbitan, sorbitol, and isosorbide present in sorbitan
mono- or tristearate can be determined using this procedure (Murohy and
Grisley, 1969; Tsuda et al., 1984). Polyols from polysorbates are detectable by
GLC, but under conditions reported by Lundquist and Meloan (1971), they are
eluted as a single peak. Detection of 1,4-dioxane, a carcinogenic byproduct of
ethoxylation reactions, may also be determined in polysorbates by GLC (Birkel
et al., 1979). Polyol distribution from diglycerol through dodecaglycerol are
obtainable for polyglycerol esters (Schuetze, 1977; Sahastrabudhe, 1967).
Polyglycerol polyricinoleate (PGPR), which has been extracted from chocolate
products, has also been determined by gas-liquid chromatography (Dick and

Jmcmal Standard

Internal Standard

PGMS

PGMP

PGMS

GMO&GMS

GMS

12

Time (min)

15

18

21

24

Time (min)

Figure 3.1 GLC separation of monoglycerides and propylene glycol ester emulsifiers.
(a) Commercial emulsifier; (b) in shortening. (Hasenhuettl et al., 1990.)

Analysis of Food Emulsifiers

57

Miserez, 1976). Although fatty acid esters of saccharides, such as galactose,


xylose, and sucrose, are unstable to high temperatures, volatile derivatives of
these esters have been determined by GLC (Tsuda and Nakanishi, 1983;
Fregapane et al., 1992). Calcium stearoyl lactylate has also been analyzed
(Yukawa and Hanada, 1982). The number of lactic acid residues esterified to
stearic acid in calcium or sodium stearoyllactylate may also be analyzed using
this procedure.

3.5.2

High-Performance Liquid Chromatography (HPLC)

As previously discussed, thin-layer and column chromatography have been


useful in the separation and analysis of lipids. HPLC may be viewed as a logical extension of these techniques. Small amounts of lipid are introduced onto a
solid or reverse phase column and the column is developed using a solvent as
a moving (mobile) phase. As peaks are eluted, they are observed and quantitated by a detector. Modern HPLC units collect data on a digital computer.
This method of analysis has an advantage over GLC because nonvolatile lipids
can be separated without the necessity of forming volatile derivatives.
However, lipids as a class pose some disadvantages for HPLC. Saturated lipids
do not absorb UV light except at short wavelengths where some useful solvents
are not transparent. Derivatives of lipids that contain chromophores can be
prepared but this negates a major advantage of HPLC. Refractive index detection may be used for detection of peaks. The analyst is then limited to an iso-

cratic solvent system because gradient elution causes drift of the IR detector.
Recently, an evaporative light-scattering detector (ELSD) has been developed
to overcome these technical problems for lipids (Christie, 1992; Bruns, I 988;
Lee et al., 1993). Figure 3.2 shows a separation of phospholipids using the
ELSD detector. HPLC can also be interfaced with an MS instrument to give
both separation and identification of peaks.
The most commonly reported separations of food emulsifiers under a potpourri of columns, solvents, and detectors have been monoglycerides (Filip
and Kleinova, 1993; Takagi and Ando, 1994; Renger and Wenz, 1989; Tajano
and Kondoh, 1987; Martin et al., 1989; Rilsom and Hoffmyer, 1978;
Brueschweiler, 1977) and phospholipids of soy (Melton, 1992; Sotirhos et al.,
1986; Hurst and Martin, 1984; Huyghebaert and Baert, 1992; Tumanaka and
Fujita, 1990; Rhee and Shin, 1982) and egg (Hsieh et al., 1981; Kaitaranta
and Bessman, 1981). Hurst and Martin (1980) have also employed HPLC to
determine phosphatidylcholine in chocolate. Separation of phospholipids has

58

Food Emulsifiers and Their Applications

Phospholipids Class Standards

Egg Yolk Phospholipids

1. Cholesterol
2. PalmHic acid
3. Phosphatidylethanolamine
4. Phosphatidylserine
5. Phosphatidylcholine
6. Sphingomyelin

12

16

1. Cholesterol
2. Phosphalidylethanolamine
3. Phosphalidylcholine
3 4. Sphingomyelin

12

2DIIin.

Column:

Spherisorb Silica, 3jlm,


100x 4.6nvn
Mobile Phase:
A: IPA, 8: Hexane, C: Waler
Gradient:
Time 0 7 15
58 52 52
A
40 40 40
8
c 2 8 8
Sample:
Phospholipids Class Slandards
Flowrate:
1.25mllmin
Detector:
ELSD Mark IIA,
Drift Tube Temp: 105'C
Gao Flow:
50mm

16

2D Min.

Column:

Spherisorb Silica, 3~m.


100x4.6mm
Mobile Phase:
A: IPA, 8: Hexane, C: Waler
Gradient:
Time o 7 15
A
585252
8
404040
c 2 8 8
Saf111le:
Egg Yolk Phospholipids
Flowrate:
1.25mUmin
Detector:
ELSD Mark IIA,
Drift Tube Temp: 105'C
Gaa Flow:
50mm

Figure 3.2 HPLC separation of phospholipids using an evaporative light-scattering


detector. (Courtesy of Alltech Associates, Inc.)

also been reviewed by Christie (1996). Garti and Aserin (1981) reported the
separation of polyglycerol mono- and polyesters by HPLC on a LiChrosorb column. Sorbitan esters of fatty acids could also be separated on the same stationary phase (Garti and Aserin, 1983). Murakami and coworkers (1989)
described the separation of sucrose esters by preparing their 3,5-dinitrobenzoyl derivatives. This technique was used to determine sucrose esters in a
wide range of food products.
The development of facile extraction and rapid sample prepurification
methods as well as autosampling technology will undoubtedly encourage more
widespread use of HPLC.

3.5.3

Spectroscopic Methods

As previously mentioned with respect to HPLC detectors, saturated lipids do


not contain chromophores that absorb in the UV range, except at short wave-

Analysis of Food Emulsifiers

59

lengths. The short-wavelength region is not selective enough to allow determination of lipids in the presence of other organic compounds. However, complexes of emulsifier functional groups have been widely utilized to analyze
emulsifiers colorimetrically. A diagnostic qualitative analysis for ionic surfactants was developed by Lew (1975). Methylene blue was used to analyze for
anionic surfactants at 650 nm while cationic compounds were determined with
Orange 2 at 485 nm.
Several determinations of phospholipids rely on complexation of the choline
moiety of phosphatidylcholine. Molybdenum blue (Hartman et al., 1980),
dipicrylamine (Mueller, 1977), and Reinecke's salt (Moelering and Bergmeyer,
1974) were used to determine lecithins in a variety of food products.
Polyoxyethylene chains in food emulsifiers can complex with cobalt thiocyanate
or Dragondorff's reagent to produce a colored complex. This method has been
used to detect polysorbates in instant noodle soup (Saito et al., 1987; Tonogau et
al., 1987) and in salad dressings (Daniels et al., 1982). Shmidt (1979) reported a
procedure for determination of DATEM esters by mixing saponified lipid with
sodium metavanadate solution and measurement of the absorption at 490 nm.
Infrared spectroscopy is applicable to the functional groups present in the
common food emulsifiers. The emergence of Fourier transform infrared spectroscopy (FTIR) makes rapid detection of functional groups possible. The technique could therefore be used to monitor manufacturing processes. Given
these capabilities, it is somewhat surprising that little work has been done to

measure emulsifiers. Ingber utilized near infrared (NIR) reflectance to determine monoglyceride concentration and determine the hydroxyl value in polyglycerols and polyglycerol esters (N. Ingber, personal communication).
Spectrophotometric analysis has also been utilized to measure impurities in
food emulsifiers. Fujita and Yamanaka (1991) measured the absorbance of iodine liberated from an iodide solution from reaction with lipid peroxides. This
technique yields equivalent information to the peroxide method described previously. Infrared absorption at 1675 cm1 was utilized to determine residual dimethylformamide in sucrose esters (Jakubska, 1977). However, this method is
inadequate to determine the solvent at ppm levels.

3.5.4

Nuclear Magnetic Resonance (NMR)

Nuclei of atoms having an odd atomic number may be measured in terms of


their chemical shifts. This measurement is carried out in an instrument that
places a sample in a magnetic field and then scans the radio frequency to ob-

60

Food Emulsifiers and Their Applications

tain signals. These signals are dependent on the environment of the nuclei and
are therefore useful for determining functional groups and chemical structure.
Relaxation time of the signal is useful to determine the physical state of a molecule. Wide-line NMR is widely used to measure the amount of solid fat or
solid fat content (SFC) in a sample. However, this technique is rarely used for
food emulsifiers and is not included here. Chemical shifts have been utilized
to determine the mesomorphic form of emulsifiers in aqueous systems, which
is likewise beyond the scope of this chapter. The reader is referred to
Lindblom (1996) for a thorough treatment of this subject. Mesomorphic phase
behavior in food emulsions will be discussed in Chapter 6.
Proton (1 H) NMR is the oldest technique. However, because of the large number of similar protons in alkyl chains of emulsifiers, this technique has not shown
widespread utility. Press and coworkers (1981) observed the protons on choline at
3.3 ppm to determine phosphatidylcholine content in lecithin. Sheeley et al.
(1986), in the same laboratory, proposed that the vinylic protons at 5.3 ppm could
serve as an alternative measurement of the iodine value, discussed earlier.

Chemical shifts of carbon (13 C) are useful for determination of the functional environment of the carbon atom being observed. Since lipids have significantly fewer carbons than hydrogen atoms, the problem of spectra

interpretation is more tractable. Gunstone (1993) has recently reviewed the


field of 13 C NMR of lipids. Chemical shifts of isopropylidene derivatives of
monoglycerides have been reported by Dawe and Wright (1988). Sacchi and
coworkers (1990) utilized the shifts of glyceryl and carbonyl carbons to measure the levels of monoglycerides, diglycerides, and free fatty acids in olive oil.
Chemical shifts for glyceryl carbon atoms are listed for monoglycerides, propylene glycol esters, acetylated monoglycerides, phosphatidylcholine, and phosphatidylethanolamine in Table 3.1.
Chemical shifts of phosphorus atoms (3 1P) are of great utility in determining
the structure and concentration in phospholipids. Since there is only one phosphorus atom per lipid molecule, assignment is more straightforward than for
1H or 13C NMR. However, this is somewhat offset by the numerous types of
phospholipid molecules found in natural samples, such as egg. Glonek and
Merchant (1996) have recently reviewed this method.

3.5.5

Mass Spectrometry

Mass spectrometry has long been used to elucidate the structures of organic
molecules. The molecular ion is an accurate indicator of the molecular weight

Analysis of Food Emulsifiers

Table 3.1

13 C

61

chemical shifts for some food emulsifiers

Emulsifier structure

Gl-1

Gl-2

GL-3

N-CH 2 O-CH 2

Soy phosphatidylcholine
Egg yolk phosphatidylcholine
Soy phosphatidylethanolamine
Egg yolk phosphatidylethanolamine
1-Monoglyceride
1,2-Diglyceride
!-Propylene glycol monoester
2-Propylene glycol monoester
Propylene glycol diester
Monoacetylated monoglyceride:

63.01
62.94
62.81
62.81
65.04
65.04
69.46
65.92
65.42

70.51
70.63
70.59
70.55
70.27
72.25
66.13

63.33
63.78
64.07
64.07
63.47
61.58
19.2 (CH 3)

66.26
66.62
40.69
40.59

71.77
67.98

16.25 (CH 3)
16.5 (CH 3)

62.07
63.00
62.00

72.39
68.19
69.16

61.40
65.26
62.33

a
~
Diacetylated monoglyceride

59.34
59.32
62.08
62.13

of the compound, while the fragmentation pattern is an indication of carbon


chain (or daughter) structure and associated functionality. Lipids and food
emulsifiers are usually nonvolatile complex mixtures of molecules of highly
varying molecular weights. Therefore, great confusion is encountered trying to
determine which species are molecular ions and which are fragmentation
peaks. New techniques such as direct sample introduction, fast atom bombardment (FAB), chemical ionization (CI), and negative ion mass spectrometry
have made analysis of lipids and emulsifiers more feasible. Tandem mass
spectroscopy (MS/MS) allows an ion peak to be further fragmented to smaller
peaks. Insight on molecular structure is therefore increased dramatically. Le
Quere (1993) has reviewed the apparatus, methodology, and application to
lipids of this technique.
Integration of chromatographic techniques, such as GLC and HPLC, can
simplify mass spectra by enabling analysis of compounds that are present in
high concentration. Application to food emulsifiers is expected to occur
rapidly as instrument and detector technology are improved.
Fast atom bombardment was utilized to analyze complex mixtures of phospholipids present in egg yolk (Traitler and Nikiforov, 1984). Protonated molecular ions (MH+) could be readily resolved and identified. Negative (OH-) chemical
ionization mass spectrometry was used to characterize polysorbates in foods
(Daniels et al., 1985). Two peak series were attributed to polyoxyethylene chains
alone and linked to sorbitans. Fatty acids attached to the sorbitans could also be

62

Food Emulsifiers and Their Applications

identified from the spectra. An electrospray technique has also been found to be
useful for analysis of sucrose fatty acid esters (Schuyl and van Platerink, 1994).
This technique showed a series of molecular ions that corresponded to the degree of substitution of fatty acids onto the sucrose molecule.

3.6

Setting Specifications for Emulsifiers

The practice of setting specifications for a food ingredient may be a matter of


custom, such as adoption of the manufacturer's values, or it may be a carefully
reasoned approach based on functionality. When food processors are making
commodities or products very similar to those on the market, the first approach
may be quite adequate and even time-saving. Emulsifier producers are very
likely to have an understanding of what the customer needs for an emulsifier to
function properly in the food system. Sometimes, however, the food processor
may develop a "new to the world" product that has no close relative in current
commerce. In this case a well-reasoned logical approach is necessary.
A logical specification approach starts with selection of an emulsifier system that imparts functionality to the finished or in-process food product. This
selection has most likely resulted from a careful regulatory and technical evaluation of possible emulsifier(s) system. During the process, the most critical
functional responses such as emulsion stability, aeration, or viscosity should
have been defined.
As a second step, each emulsifier in the system should be correlated to
each functional response.
Third, critical analytical values need to be selected for each emulsifier.
Most of these tests have been discussed, but which of them show a correlation
with functionality is not always clear. A factorial design may be constructed for
each analytical constant and each emulsifier as measured by the critical response function. Emulsifiers having a range of analytical values should be
tested. If a wide range is not available, an approximation could be obtained using a low and high lot and preparing blends to represent values in the middle.
Based on the analytical constant correlation with function, a range of values
is selected that give acceptable functional response. A statistical safety factor
should be defined and specifications appropriately tightened.
As a reality check, the emulsifier manufacturer and food processor should evaluate the specifications to determine whether they can realistically be met. If not,
further experimental design may be necessary to arrive at a more robust system.

Analysis of Food Emulsifiers

63

Finally, appropriate purity analyses should be defined. These are analyses


such as color, odor, and peroxide value that can influence the appearance, flavor, and shelf stability of the finished food product. For example, an unsaturated monoglyceride may need to have a low peroxide value to keep it from
acting as a pro-oxidant in a susceptible product.

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Food Emulsifiers and Their Applications

Filip, V., Kleinova, M.(l993). Z. Lebensm.-Unteres. Forsch., 196(6):532-5.


Firestone, D. (1994). ]. Assoc. Off. Anal. Chem., 77(3):677-80.
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Ibid. (1990b). Method Cc 13b.
Ibid. (1990c). Method Cc 13d-55.
Ibid. (1990d). Method Cc 14--59.
Ibid. (1990e). Method Cc 14--59.
Ibid. (1990f). Methods Cd 1-25, Cd 1b-97.
Ibid. (1990g). Method Cd 1b--9l.
Ibid. (1990h). Method Cd 1c-85.
Ibid. (1990i). Methods Cd 2a-45, Ja 2a-46.
Ibid. (1990j). Methods Cd 3-25, Cd 3b--76, Cd 3c9l.
Ibid. (1990k). Methods Cd 3a-63, Cd 2d-63, Ja 6-55.
Ibid. (19901). Method Cd 4-40.
Ibid. (1990m). Method Cd 5-40.
Ibid. (1990n). Method Cd 11-57.
Ibid. (1990o). Method Ce 1b-89.
Ibid. (1990p). Method Ja 2b-87.
Ibid. (1990q). Method Ja 4-46.
Ibid. (1990r). Method Ja 5-55.
Ibid. (1990s). Method Ja 8-87.
Ibid. (1990t). Method Ja 9-87.
Ibid. (1990u). Method Ja 10-87.
Ibid. (1990v). Method Ja 11-87.
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Franzke, C., Kroll, 1. (1980). Nahmng, 24(1):89-90.
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Fregapane, G., et al. (1992). Prog. Biotechnol., 8:563-8.
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Goldstein, S. (1984). U.S. Patent 4,473,651 A, Sept. 25.
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Lipid Methodology -Two (ed. W.W. Christie), The Oily Press, Ayr, Scotland.
Halkier, S.B., (1980). Ger. Int. Ges. Getreidechem., 10, 36-7.
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Hasenhuettl, G., et al. (1990). ]. Am. Oil Chem. Soc., 67(11):797-9.
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Hsieh, J.Y., et al. (1981). ]. Chromatogr., 208(2):398-403.
Hurst, W.J., Martin, R.A. (1984).]. Am. Oil Chem. Soc., 61(9):1462-3.

Analysis of Food Emulsifiers

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Huyghebaert, G., Baert, L. (1992). Chromatographia, 34(11-12):557-62.
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FOUR

Carbohydrate/Emulsifier
Interactions
Lynn B. Deffenbaugh

4.1

Introduction

Many different types of emulsifiers are used in food products. The inclusion rate is
usually low, < 3%. Composition and characteristics of emulsifiers used in foods
are discussed in other chapters. Emulsifiers often function indirectly in foods by
modifying the properties of major components such as carbohydrates, protein, or
fat. This chapter discusses the impact of emulsifiers on carbohydrates.
Carbohydrates are a large group of diverse food components. Carbohydrates
can be classified into three categories based on degree of polymerization: monoand disaccharides, oligosaccharides, and polysaccharides. Polysaccharides
are further divided into starch and nonstarch fractions. The nonstarch polysaccharides are plant cell wall materials that contribute dietary fiber to foods.
Mono-, di-, and oligosaccharides and nonstarch polysaccharides (fiber) interact minimally with emulsifiers from the perspective of food-product characteristics. Starches, however, form complexes with emulsifiers that significantly
alter the characteristics that the starches provide in foods. Emulsifiers that
complex with starch are notably different in function and usage from proteincomplexing emulsifiers discussed in the next chapter.
This chapter discusses how complexes form between starches and emulsi-

67

68

Food Emulsifiers and Their Applications

fiers, and how starch properties are affected by complex formation.


Understanding the effects of emulsifiers on carbohydrates is dependent on
measurement of physical properties of starch/emulsifier complexes, which are
also discussed.

4.2

Inclusion Complexes of Starch

The interaction between emulsifiers and starches is primarily formation of an


inclusion or clathrate complex. Linear sections of starch molecules can form a
helix into which a fatty adjunct can enter. The lipophilic portion of an emulsifier is often a fatty acid chain connected to a hydrophilic portion via an ester
linkage. The fatty acid chain can serve as the fatty adjunct in an inclusion
complex with starch.
Starches are composed of linear molecules (amylose), and branched molecules (amylopectin). In an inclusion complex with amylose, the amylose exists
as a left-handed helix with 6 glucosyl residues per turn and 0.8 nm between
helices (Mikus et al., 1946). The branch points of amylopectin intercept helix
formation and limit complex formation. Monoglycerides, one of the most commonly used types of emulsifiers, are known to bind with amylopectin at much

lower levels than with amylose (Hahn and Hood, 1987; Lagendijk and
Pennings, 1970; Twillman and White, 1988).
In general, a complexing agent can be any hydrophobic molecule with a rodlike shape and a diameter less than the 4.5 to 6 A-wide internal core of a starch
helix (Rutschmann and Solms, 1989). Complexing agents include iodine (asP-),
fatty acids, hydrocarbon chains of emulsifiers, and various organic molecules
(i.e., n-butyl alcohol, t-butyl alcohol, dimethyl sulfoxide, and a-naphthol).
Organic aroma compounds have been shown to form thermostable inclusion
complexes with amylose (Maier et al., 1987; Schmidt and Maier, 1987).
Amylose alone exists in solution as a random coil. In the presence of complexing agents, energy minimization drives amylose into a helix configuration
(Neszmelyi et al., 1987). Hydrocarbon chains of fatty acids or hydrophobic
complexing adjuncts are attracted into the hydrogen-lined, hydrophobic core
of an amylose helix (Krog, 1971). Once formed, the complex is stabilized by
dipolar interactions (Mikus et al., 1946) and fulfillment of hydrophobic solvation requirements of the helix (Krog, 1971). Computer-derived models of amylose complexes with stearic acid and iodine have been derived on the basis of
the energy minimization concept (Neszmelyi et al., 1987).

Carbohydrate/Emulsifier Interactions

69

The long axis of complexing agents is internal and parallel to the long axis
of the amylose molecule (Mikus et al., 1946; Rundle and French, 1943a, b).
Complexing agents interchange reversibly, competing for the same space in
the amylose helix (Mikus et al., 1946; Schoch and Williams, 1944). The unit
cell packing size and distance between starch helices are not altered by the
type of complexing agent (Raphaelides and Karkalas, 1988).
Lipid chains usually occupy the helix as paired dimers linked by hydrogen
bonds at the carboxyl groups in fatty acids or glycerol in monoglycerides
(Raphaelides and Karkalas, 1988). The hydrocarbon chains in complexes are so
highly ordered that they have been reported to be crystalline (Carlson et al., 1979).
Amylose helices induced by complexation aggregate into partially crystalline structures that exhibit a V-type x-ray diffraction pattern (Szczodrak and
Pomeranz, 1992) and are insoluble. These insoluble complexes consist of
lamellar crystals with the helix perpendicular to the lamella (Raphaelides and
Karkalas, 1988). Amylose and amylopectin complexes can be differentiated by
their physical properties. Amylopectin emulsifier complexes are more soluble
than amylose complexes. The insolubility of the fatty acid/amylose complex
has long been used as a means to selectively precipitate amylose, but not amylopectin, from solution (Schoch and Williams, 1944). Complexes cannot always be assumed to be insoluble, however, since the degree of insolubility
varies with the surfactant (Kim and Robinson, 1979). The complex between
starch molecules and iodine is used to differentiate and quantitate the amylose
fraction, which forms a blue complex, and the amylopectin fraction, which
forms a red-purple complex.

4.3 Effects of External Lipid Materials


on Starch Properties
4.3.1 General
Either native or external lipid materials can interact with starch. The mechanisms and effects of the interactions are qualitatively similar but much larger in
magnitude for external lipids such as emulsifiers added to foods. While native
lipids significantly affect starch properties, the presence and level is not discretionary in product development. The effect of emulsifiers on starch properties in food is especially relevant because use of emulsifiers provides choice
and control in product development. This chapter focuses on the effects of externally added emulsifiers on starch properties.

70

Food Emulsifiers and Their Applications

Many food emulsifiers function by interacting with the starch component of


food. Emulsifiers were first used as crumb softeners in breads and were shown
to interact with starch but not gluten (Favor and Johnston, 1947; Strandine et
al., 1951). Starch-complexing emulsifiers are used to retard firming or staling
in bread, prevent stickiness in instant mashed potatoes, and control texture in
extruded products.
Specific effects of emulsifiers and various fatty adjuncts on starch properties are described in the accompanying tables. Data for nonwaxy "normal"
starches are summarized in Table 4.1, and data for waxy, high amylopectin
starches are summarized in Table 4.2. These data from many sources are consistent overall and support general conclusions about starch/emulsifier interactions, as discussed below.

4.3.2

Iodine-Binding Capacity

Emulsifiers or externally added fatty adjuncts repress iodine binding capacity


(IBC) of nonwaxy starches (Table 4.2). This effect on IBC is caused by the reversible exchange between the adjunct and J3- within the starch helix.
No or only slight reduction of IBC with waxy starches or amylopectin have
been reported (fable 4.2). The average length of amylopectin branches is 20 to
26 residues. Fatty acids require 3 turns of a starch helix with 6 residues per
tum to form a complex, and thus can interact with amylopectin (Rundle and
French, 1943b). The IBC of amylopectin is low compared to normal or high
amylose starches, and starch-complexing agents decrease IBC of waxy maize
starch slightly (Figure 4.1). Formation of weak or unstable complexes between
amylopectin and emulsifiers may significantly modify the starch properties,
but IBC is not a sensitive method to predict or monitor the interaction.

4.3.3

Starch-Pasting

Starches and starch-containing ingredients provide many of the physical and


organoleptic properties of foods. Dramatic changes occur in starches and
starch-containing products when processed. When starches are cooked in the
presence of water, starch granules absorb water and swell to many times their
original size. Amylose, the linear fraction of starch, leaches from the granules.
A hot starch paste is a mixture of swollen granules, granule fragments, and colloidally and molecularly dispersed starch molecules (Olkku and Rha, 1978).
The viscosity increases greatly as this hot paste matrix forms. The fragile,
swollen granules will eventually disintegrate, especially when shear is ap-

......
......

Sucrose monoesters
GMS
Sucrose ester
GMS, SSL
EMG, polysorbate 60
MG
CTAB, GMS, SLS
Sucrose esters
Sucrose monoesters
GMS, SSL
GMS, SSL
MG
EMG, polysorbate 60
MG
MG
CTAB, GMS, SLS
Sucrose esters
MG
MG,SSL
MG

Wheat
Potato
Maize, potato, tapioca, wheat
Wheat
Potato amylose
Amylose

Tapioca
Amylose
Wheat
Maize, potato, wheat
Wheat
Potato
Potato amylose
Amylose
Tapioca
Tapioca
Amylose
Nonwaxy
Wheat flour
Nonwaxy

Reduce iodine-binding capacity

Repress granule swelling


and starch solubilization

Complexing
agent

Starch
type/fraction

Effect of emulsifiers and complexing agents on properties of nonwaxy starch

Effect of complexation
on starch properties

Table 4.1

Bourne et al., 1960


Conde-Petit and Escher, 1992
Deffenbaugh, 1990a
Ghiasi et al., 1982a
Kim & Robinson, 1979
Krog, 1971
Krog & Nybo Jensen, 1970
Moorthy, 1985
Osman et al., 1961
Bourne et al., 1960
Eliasson, 1986b
Ghiasi et al., l982a
Hoover & Hadziyev, 1981
Kim & Robinson, 1979
Krog, 1971
Mercier et al., 1980
Moorthy, 1985
Osman et al., 1961
Strandine et al., 1951
Roach & Hoseney, 1995a,b
van Lonkhuysen & Blankestijn,
1974

Reference(s)

.....
I()

Sucrose ester
Sucrose monoesters
SSL
SSL
DATEM, MG, SSL
POEMS
MG
DATEM, MG, SSL

DATEM, MG, SSL


GMS, SSL
MG
Sucrose monoesters
Sucrose monoesters
MG,SSL
GMS, SDS, SSL
MG, SSL
Sucrose monoesters
MG
MG

Wheat
Wheat
Potato
Wheat
Wheat starch
Wheat
Maize, potato, wheat
Wheat
Wheat flour
Potato
Various

Maize, potato, tapioca, wheat


Wheat starch
Wheat
Potato, wheat
Wheat
Nonwaxy
Maize
Maize, potato, tapioca, wheat

SLS, an anionic surfactant

Tapioca

Increase initial pasting


temperature, hot paste
viscosity, temperature of
peak viscosity Walker, 1984
(i.e., amylograph or RVA),
delay gelatinization

SDS, an anionic surfactant

Maize, potato, wheat

Increase granule swelling;


make gelatinization occur
earlier
Destabilize granule and
increase paste viscosity
Decrease starch thickening
power < 85C (before
gelatinization)
Delay loss of
birefringence

Complexing
agent

Starch
type/fraction

continued

Effect of complexation
on starch properties

Table 4.1

Evans 1986
Ghiasi et al., 1982d
Hoover & Hadziyev, 1981
Bourne et a!., 1960
Ebeler & Walker, 1984
Eliasson, 1985
Eliasson, 1986b
Ghiasi et a!., 1982a,b
Pomeranz et a!., 1969
Rilsom eta!., 1984
van Lonkyhuysen &
Blankestijn, 1974
Deffenbaugh, 1990a
Ebeler & Walker, 1984
Eliasson, 1983
Eliasson, 1986b
Evans, 1986
Favor & Johnston, 1947
Krog, 1971
Krog, 1973

Moorthy, 1985

Eliasson, 1986b

Reference(s)

.....
w

Decrease retrogradation of starch

Increase setback viscosity


Increase setback viscosity (gelation)
Depressed G' and G"; increased
temperature of G' and G";
increased viscous part of
viscoelastic response
Induced gelation (increased
rigidity of fresh starch gels)
Decreased gel volume of
heated starch
Decrease cold paste viscosity

Stabilize pasting viscosity and


prevent long cohesive texture
Decrease peak viscosity
Decrease gelatinization enthalpy
Eliasson, 1986a
Twillman & White, 1988
Deffenbaugh, 1990a
Eliasson, l986b

Conde-Petit and Escher, 1994


Eliasson, 1985
Favor & Johnston, 1947
Krog, 1971
Osman & Dix, 1960
Hoover & Hadziyev, 1981
Riisom eta!., 1984
Gudmundsson & Eliasson, 1990
Krog & Nybo-Jensen, 1970
Lagendijk & Penning, 1970

C'fAB, sat'd MG, SDS, SSL,


Lecithin, lysolecithin
MG
Sucrose ester
GMS, SLS

CSL, GMS
MG, SSL
POEMS
MG
MG
MG
CTAB, SDS

Maize, potato, wheat

Wheat

Maize, potato, tapioca, wheat


Maize

Potato
Potato
Amylose/amylopectin mixtures

Masa harina flour


Maize, potato, tapioca, wheat
Maize, potato, wheat

Favor & Johnston, 1947

POEMS
Sucrose ester

waxy maize, potato


Maize, potato, tapioca, wheat
Deffenbaugh, 1990a
Potato, wheat

Lin et al., 1990


Pomeranz et a!., 1969
Riisom et a!., 1984
Twillman & White, 1988
Moorthy, 1985

SSL
Sucrose monoesters
MG
MG
GMS, SLS

Pea flour
Wheat flour
Potato
Masa harina flour
Tapioca

....

continued
Complexing
agent

DATEM, MG, SSL,


Sucrose esters
Sucrose esters
EMG, DATEM, MG, SSL
CSL,GMS
MG
CSL, MG
MG, SSL
EMG, polysorbate 60
MG
MG
Lysolecithin
Sucrose ester
Lysolecithin
Lysolecithin

Starch
type/fraction

Rice

Maize

Barley, maize, waxy maize

Maize, potato, wheat


Tapioca
Potato and maize
Potato
Potato amylose
Amylose

Amylose
Potato amylose
Maize, potato, tapioca, wheat
Potato amylose
Potato amylose

Eliasson & Krog, 1985


Holm et al., 1983
Deffenbaugh, 1990a
Holm et al., 1983
Holm et al., 1983

Conde-Petit and Escher, 1994


Mercier et al., 1980
Staeger et al., 1988
Ghiasi et al., i982
Kim & Robinson, 1979
Eliasson & Krog, 1985

Szczodrak & Pomeranz, 1992

Matsunaga & Kainuma, 1986

Miura et al., 1992

Reference(s)

CSL =calcium stearoyl-lactyl-2-lactylate; CTAB =cetyltrimethylammonium bromide; DATEM =diacetyl tartaric acid esters of monoglylcerides; EMG =ethoxylated
monoglycerides; GMP =glycerol monopalmitate; GMS =glycerol monostearate; MG =monoglycerides; POEMS =polyoxyethylene stearate; SDS =sodium dodecyl
sulfate; SLS =sodium Iaury! sulfate; SSL =sodium stearoyl-2-lactylate.

Decrease amylopectin
recrystallization
Decreased formulation of
resistant starch
Reduced gel breaking strength
Reduced starch extrudate
solubility and retrogradation
Reduced in vitro enzymolysis
with B-amylase
Reduced in vitro amyloglucosidase
digestion
Reduced in vitro a-amylase
digestion
Decreased glucoamylase
digestibility
Slowed rate of in vivo a-amylase
digestion

Effect of complexation
on starch properties

Table 4.1

.......

Ul

Bourne et al., 1960


Batres & White, 1986
Staeger et al., 1988
Deffenbaugh, 1990a

Sucrose monostearate
Sucrose esters
MG
DATEM, MG, SSL
POEMS
GMS, SLS

Sucrose monostearate
MG
CSL, MG
Sucrose esters

Potato amylopectin
Waxy maize
Amylopectin
Waxy maize
Waxy maize
Waxy barley

Potato amylopectin
Amylopectin
Waxy maize
Waxy maize

No extrudate complex formed


No complex detected by x-ray
diffraction or DSC

Decreased hot paste viscosity


Depressed G' and G"; slightly
increased temperature of G'
and G"; slightly increased viscous
part of viscoelastic response
Insoluble complex precipitated

Bourne et al., 1960


Deffenbaugh, 1990a
Hoover & Hadziyev, 1981
Evans, 1986
Favor & Johnston, 1947
Eliasson, 1986b

Sucrose esters
Sucrose monostearate

Waxy maize
Potato amylopectin

Deffenbaugh, 1990a
Bourne et al., 1960

Krog, 1971; Krog & Nybo-Jensen,

MG

Amylopectin

Reference(s)

Slight reduction iodine1970


binding capacity
No reduction in iodinebinding capacity
No effect on swelling
Slight delay in peak viscosity
Viscosity profile not affected

Complexing
agent

Starch
type/fraction

Effect of emulsifiers and complexing agents on properties of waxy starch

Effect of complexation
on starch properties

Table 4.2

Gudmundsson & Eliasson, 1990


Eliasson, 1988
Gudmundsson & Eliasson, 1990

CTAB, SDS
CTAB, unsat'd MG
CTAB, SDS

Potato amylopectin

Waxy maize, amylopectin


Potato amylopectin

CSL = calcium stearoyl-lactyl-2-lactylate; CTAB = cetyltrimethylammonium bromide; DATEM = diacetyl tartaric acid esters of monoglycerides; GMS = glycerol
monostearate; MG = monoglycerides; POEMS= polyoxyethylene stearate; SDS =sodium dodecyl sulfate; SLS =sodium Iaury! sulfate; SSL =sodium stearoyl-2-lactylate.

Deffenbaugh, 1990a

Sucrose esters

Waxy maize

Reference(s)

Weak complex suggested by


glucoamylase digestion, viscosity
profiles, high-performance size
exclusion chromatography,
and NMR
Complex confirmed by DSC
and x-ray diffraction
Reduced amylopectin
retrogradation

Complexing
agent

Starch
type/fraction

continued

Effect of complexation
on starch properties

Table 4.2

Carbohydrate/Emulsifier Interactions

.--..

77

1.0

...::::
()
L..

.....0en

0.8

--Hylon VII
o--oWheat
A - - A Maize
c--o Potato
o--oTapioca
v--vWaxy Maize

0>

0.6
IJ")
...._,

c:
0
0

tO
.....
0

en

0.4
0.2

..0
<(

0.0

T--------v-----~v------------------------------v

2
3
% SE (starch wt. basis)

Figure 4.1 Iodine-binding capacity of starches measured in the presence of a sucrose


ester emulsifier. (From Deffenbaugh, 1990a.)

plied. This relative decrease in viscosity after time and application of shear
defines a peak viscosity.
The process whereby the ordered structure of the starch granule is lost is
called "gelatinization." The transition is a first-order water-mediated melting
of crystalline regions in the starch granule (Donovan, 1979; Zobel, 1984).
Maximum solubilization and swelling occur when excess (> 5 times) water is
present, typical of sauces, gravies, and puddings. In lower moisture systems,
such as baked or extruded products, granule swelling can be limited by moisture content. Extremely high viscosities result because diluent is limited.
Uses and functions of starches are multiplied by modifying properties with
emulsifiers. For example, time of addition and/or time for diffusion of fatty adjuncts into starch granules are processing parameters than can be varied to
produce substantially different properties in cooked starch or cereal grain
products (Lund, 1984). If added before starch gelatinization, monoglycerides
penetrate the granule, form molecular complexes, and reduce starch swelling
power. Addition of monoglycerides after gelatinization enhances granule stability (van Lonkhuysen and Blankestijn, 1974). The function of starch-complexing emulsifiers is primarily due to formation of starch/emulsifier
complexes. The magnitude of the effect of starch/emulsifier complexes on
starch cooking properties varies with starch type, emulsifier type, and processing conditions.

78

Food Emulsifiers and Their Applications

Emulsifier and starch molecules must have access to each other to interact.
Starch-complexing emulsifiers must be soluble enough or in a phase that
yields monomers (see Section 4.4.4). Starch molecules have limited availability until gelatinization begins. Emulsifiers adhere to the granule surface before
gelatinization, and they begin to form insoluble starch/emulsifier complexes as
soon as the granule begins to swell and amylose begins to solubilize. These insoluble complexes near the surface stabilize the granule. The rate of further
swelling and amylose leaching are slowed. Gelatinization temperature is increased because more energy is required to cook or swell the stabilized granule. Some emulsifiers, such as polysorbate 60, may cover the starch surface
with a film, increase hydrophobicity, and inhibit water transfer into the granule
(Kim and Walker, 1992c).
The effect of emulsifiers on starch gelatinization is detected in many ways
(Table 4.1). When starch pastes were made with SSL or GMS, changes in viscoelastic properties of the paste were coincident with reduced granule swelling
(Eliasson, 1986b). The granules were less deformable (stiffer), as indicated by
an increase in the temperatures where peaks in storage (G') and loss (G") module were reached. Pasting temperature, hot viscosity, and temperature of peak
viscosity of most normal starches are increased in the presence of emulsifiers
that have a fatty adjunct that can form inclusion complexes with starch.
According to Mitchell and Zillman (1951), the increase in starch paste viscosity in the presence of complexing agents is a result of the increased ability of
granules to absorb and hold water without rupturing.
Gelatinization, the melting of crystalline regions in the granule, causes loss
of birefringence and disappearance of a starch x-ray diffraction pattern
(Eliasson, 1986a). When emulsifiers delay gelatinization, the loss of birefringence is delayed (see Table 4.1).
Some emulsifiers with a fatty acid chain adjunct do not form complexes
with starch. Anionic surfactants, such as sodium lauryl sulfate (SLS) and
sodium dodecyl sulfate (SDS), destabilize the granule because of its highly
negative charge (Eliasson, 1986b; Moorthy, 1985). The destabilization leads to
rapid, complete swelling of the granules, and a rapid increase in viscosity
early in the pasting (cooking) cycle. Loss of granule integrity will subsequently
lead to a dramatic loss in viscosity after peak. This is undesirable if the starch
is being utilized for its thickening, texturizing abilities. In the selection of an
emulsifier for product development applications, charge as well as chemical
composition need to be considered when effect on starch is important.

Carbohydrate/Emulsifier Interactions

4.3.4

79

Starch Paste Gelation

The formation of a composite starch gel is responsible for the texture and
structure of many food systems. A starch gel formed from a paste is a composite of swollen starch granules embedded in and reinforcing the amylose gel
matrix (Ring, 1985). As a starch paste cools, molecules become less soluble
and aggregate, and a three-dimensional gel forms (Osman, 1967). The consistency increases because molecular associations form a cross-linked network
that increases the resistance of the paste to deformation (Zobel, 1984).
Gelation is initiated by rapid precipitation of amylose in solution, while amylopectin gels much slower and requires higher concentrations. Amylose serves
as (i) the chief material for formation of a gel network to entrap unabsorbed water and (ii) a binding material to link intact or fragmented swollen granules.
In starch gels made with emulsifiers, the insoluble complex produced will
form a gel (Conde-Petit and Escher, 1992). The formation of complexes between amylose and emulsifiers accelerates gelation in the first few hours of aging after gelatinization compared to starch gels made without emulsifiers
(Conde-Petit and Escher, 1994). The gelation of maize, potato, tapioca, and
wheat starch during cooling is responsible for the setback viscosity of profiles
shown in Figure 4.2 (Deffenbaugh, 1990a). A sucrose ester emulsifier increased setback viscosity due to complex formation that accelerated gelation.
Gelation can be induced and controlled by use of emulsifiers in starch-containing foods (Conde-Petit and Escher, 1992).

4.3.5

Retrogradation

The primary reasons for using emulsifiers are modification of finished product
characteristics. Retrogradation in starch-containing foods impairs texture and
taste, and emulsifiers inhibit this retrogradation.
Retrogradation is the formation of ordered, partially crystalline sites within
a cooled starch paste or gel. The recrystallization of starch is a long-term
process that occurs during the hours and weeks after pasting and gelation.
Amylose retrogrades so fast that it is often complete before the food is consumed. Retrogradation of amylopectin occurs more slowly and affects the texture and taste offoods throughout shelf life (Miles et al., 1985).
Emulsifiers decrease the rate of retrogradation of starch gels mainly through
emulsifier/starch-complex formation (Miura et al., 1992). The helix structure of
starch in a complex prevents side-by-side stacking of starch molecules and,

80

Food Emulsifiers and Their Applications

200

100

100

- MIn
- l S Sit
-2!1. 5I
pp

gao

Sl

];-

'
5
~

1;-

60

40

Ill

-~

40 .

I5>0CI

B
I 0 IZ
Time (n1in)

H.ot to g~0c

I<

16

18

20

100

1:-

8 60
40

...0

40

20

0
~ 20
0~~-+--~+--l~r-- ~--

I50CI

Heat to

10

IZ

Time (min}
95C
I

14

Cool

16

to

18

20

~C

: h

- w,.,.. .

~: ~~

eo
GO

.'-

~il

16

--

IB

20

c~ lo S0C

100

-1"-H

~~--~-+--~~-+_,

120

-Phl
I:IL $(

0
~ 20

~ 20
0

60

/ - - - - - -

;~\:...

~~/----~

;.\\ \..

//1

\~:~. ]!

1 i:
I i:

,_{.~-!:,'

10

12

Timl!!(mln}

Heat to 95C

14

IG

18

Cool lo .50C

Figure 4.2 Rapid Visco Analyzer viscosity profiles of maize, potato, tapioca, and
wheat starches with 0, 1, 2, or 5% (starch wt basis) of a sucrose ester emulsifier. (From
Deffenbaugh, J990a .)

thus, reduces nucleation sites for retrogradation or recrystallization (Matsunaga


and Kainuma, 1986).
Amylopectin retrogradation plays an important role in the shelf life of
foods. The increase in firmness and loss of flavor in staled bread is caused by
the retrogradation of the amylopectin fraction of starch (Schoch and French,
1947). Controlling or modifying amylopectin retrogradation through use of
emulsifiers is of practical significance.
The interaction between emulsifiers and amylopectin is harder to prove
than the interaction between emulsifiers and amylose. In the literature, indirect or inconclusive evidence of amylopectin/emulsifier interactions have been
reported by various authors (Kugimiya et al., 1980; Evans, 1986; Eliasson and
Ljunger, 1988). For example, formation of insoluble complexes between a
monoglyceride and amylopectin has been reported (Batres and White, 1986).

20

Carbohydrate/Emulsifier Interactions

81

A weak, limited interaction between amylopectin and a sucrose ester emulsifier was observed with various methods (Deffenbaugh, 1990a). IBC and glucoamylase digestion of waxy maize starch were slightly reduced by the
emulsifier. Delay in development of viscosity during gelatinization of waxy
maize starch also suggested that the emulsifier interacted with amylopectin.
Conclusive data describing direct and indirect effects of emulsifiers on amylopectin retrogradation are now available (Gudmundsson and Eliasson, 1990).
Emulsifiers were found to be most effective at directly inhibiting amylopectin retrogradation at very high, near 100%, amylopectin levels. Amylopectin/emulsifier
complexes were responsible for this direct effect. In the presence of amylose,
amylopectin will cocrystallize with amylose, which forms nuclei. When amylose
and amylopectin are present together, ligands will preferentially complex with
amylose. The amylose then cannot co-crystallize with amylopectin, and the emulsifier's effect on amylopectin is indirect. In addition, when the amylose component complexes with emulsifiers, less emulsifier is available to interact with
amylopectin, and direct amylopectin/ emulsifier-complex formation is reduced.

4.3.6

Enzymolysis

Complex formation interferes with the ability of enzymes to digest starch molecules
(see Table 4.1). Kinetics of glucoamylase, which cleaves successive glucose units
from the nonreducing end of starch chains, were not directly affected by a sucrose
ester emulsifier (Deffenbaugh, 1990a). Reduced digestibility of normal starches in
the presence of an emulsifier were due to decreased availability of the starch substrate. Precipitation of the complex and reduced steric compatibility of helical
starch with the active site of the enzyme reduced the starch substrate availability.
Glucoamylase digestion of waxy maize starch was not inhibited by a sucrose ester (Deffenbaugh, 1990a). If a complex formed in the outer branches, it
was not stable enough or strong enough to inhibit glucoamylase.
The inhibition of starch enzymolysis by emulsifiers is analytical evidence of
complex formation. It should be noted however, that emulsifier/starch-complex
formation does not have a significant effect on overall extent of digestibility of
starch in vivo as shown in rats (Holm et al., 1933).

4.4

Factors Affecting Complex Formation

4.4.1

Adjunct Properties

Binding of saturated fatty acids and monoglycerides increases as chain length


increases (Hahn and Hood, 1937; Gray and Schoch, 1962). Maximum binding

82

Food Emulsifiers and Their Applications

has been reported with 18-carbon saturated fatty acids (Hahn and Hood, 1987)
and 14- to 16-carbon fatty acids on monoglycerides (Hoover and Hadziyev,
1981; Krog, 1971; Lagendijk and Pennings, 1970). Geometrical structure as
well as chain length and molecular weight will influence the ability of the ligand to penetrate the starch helix (Miura et al., 1992). If chain length as well as
helix conformation are known, the fatty acid to amylose ratio required to
achieve saturation of the amylose helix is predictable from stoichiometric relationships (Karkalas and Raphaelides, 1986).
Solubility of the adjunct affects the equilibrium between complex formation
and concentration of the adjunct in solution. The lower solubility of fatty acids
versus monoglycerides in aqueous systems drives fatty acids into the lipophilic
core of the amylose helix more than monoglycerides.
Increased unsaturation in free fatty acids or fatty acid chains of monoglycerides reduces the ability of the adjunct to bind with starch (Hahn and Hood,
1987; Krog, 1971; Lagendijk and Pennings, 1970). The 30 bend at cis bonds
inhibits rotation and causes steric hindrance. In addition, lower solubility of saturated fatty acids will favor complex formation with the hydrophobic helix core.
Steric hindrance from bulky groups on emulsifiers and surfactants interferes
with the ability of the adjuncts to form complexes with starch (Gray and Schoch,
1962). Monoglyceride binding is less than binding of fatty acids of the same
chain length because steric hindrance of the glycerol group interferes with complex formation (Hahn and Hood, 1987). Propylene glycol esters of fatty acids,
with a particularly bulky polar group, have lower complexing ability than emulsifiers with smaller polar groups (Krog, 1971). In addition to steric hindrance,
hydrophilic polar groups are not compatible with the hydrophobic helix core.
The higher the proportion of the polar group to the overall molecule, the less
compatible the molecule is with the hydrophobic interior of the helix.

4.4.2

Starch Granule

The starch granules may also sterically hinder starch complexation with adjuncts in solution. Monoglycerides exist in micelles in solution and attach to
the starch granule surface simply by adsorption at low temperatures (< 50C)
(van Lonkhuysen and Blankestijn, 1974). As the temperature increases, the
monoglycerides penetrate swollen granules to complex with amylose. Other
workers measured strong surfactant/starch complexes at temperatures as low
as 60C, where only slight disorganization (gelatinization) of the starch granule
has occurred (Ghiasi et al., 1982c). The amount of the insoluble, possibly crys-

Carbohydrate/Emulsifier Interactions

83

talline complex, increased when held at 60C for 1 hour. These authors concluded that the surfactants do enter the granule and complex with amylose before gelatinization.

4.4.3

Starch Type

The source of starch in part determines its functionality in foods, and different
starches will be affected by emulsifiers in different ways and to varying extents. GMS restricted swelling of potato starch granules more than it did maize
or wheat starch granules (Eliasson, 1986b). Some methods of analysis, such as
iodine-binding capacity and glucoamylase digestion, are less sensitive to the
unique reaction of different starches in the presence of an emulsifier
(Deffenbaugh, 1990a). Other methods, such as viscoelastic properties of starch
gels (Eliasson, 1986b) and viscosity profiles (Deffenbaugh, 1990a) do differentiate the response of different starches in the presence of emulsifiers.
Viscosity parameters of various starches in the presence of a sucrose ester
emulsifier are shown in Table 4.3 (Deffenbaugh, 1990a). Time to peak viscosity was changed more in tapioca than in maize, wheat, and potato starches. The
emulsifier changed peak viscosity most in potato starch, and changed setback
viscosity most in wheat starch. Potato and tapioca granules were stabilized by
complex formation so that granule swelling and disintegration were more gradual. Moorthy (1985) also reported that starch-complexing emulsifiers stabilized the pasting viscosity of tapioca starch. Viscosity profiles are a
convenient, easy way to study complex interactions between components in
food systems.

4.4.4

Environmental Conditions

Iodine and fatty acid-binding capacity of amylose decrease with increasing


temperature (Banks and Greenwood, 1975; Hahn and Hood, 1987). Thermal
stress reduces the strength of interactions holding the complex together, and
the helix becomes more disorganized as temperature increases.
Binding of some fatty acids is affected by pH via changes in protonation of
the carbonyl group (Hahn and Hood, 1987). Palmitic or stearic acid form dimers
near or below the pKa (pH 4. 7 to 5.0) where the carbonyl groups are partially
protonated. It is assumed that dimers form by parallel alignment, the double hydrocarbon chain cannot fit into the amylose helix. Above the pK"' dimers dissociate and complex formation with single fatty acid chains occurs. pH does not
affect binding of fatty acids, such as lauric or myristic acid, that do not form

84

Food Emulsifiers and Their Applications

Table 4.3 Viscosity profile parameters of starches with 0, 1, 2, or 5% (starch wt


basis) sucrose ester emulsifier. (From Deffenbaugh, 1990a.)

T'lllle to peak (min)


Starch

0% SE

1% SE

2% SE

5% SE

Maize
Potato
Tapioca
Wheat

5.431
3.031
3.67 1
7.32 1

5.962
3.642
4.262
8.082

6.72 3
4.083
7.23 3
8.453

7.724
5.154
8.33 4
8.844

Waxy maize

3.45 1

3.54 1

3.862

4.163

Starch

0% SE

1% SE

2%SE

5% SE

Maize
Potato
Tapioca
101.3223
Wheat

57.9 1
256 1
113.2 1

77.2 2
2322
104.92

74.3 2
2263
99.63

65.93
183.64

78.4 1

80.11

81.2 1

81.61

Waxy maize

88.8

101.62

98.02

89.83

Starch

0% SE

I% SE

2% SE

5% SE

Maize
Potato
Tapioca
Wheat
Waxy maize

55.0 1
83.9 1
61.5 1
78.81 1
50.2 1

86.02
83.9 1
68.12
90.92
51.0 1

93.83
110.32
84.83
129.!3
52.61

97.4 3

Peak viscosity (%)

Maximum setback viscosity (%)

I, 2, 3, 4

118.04
166.64
51.31

These numbers indicate significant difference (P < 0.05) within starch type.

dimers at lower pH. pH does not affect binding of monoglycerides because the
carbonyl group of the fatty acid is involved in the ester linkage to glycerol.
Amylose-complexing ability of compounds containing hydrocarbon chains
is significantly affected by the phase behavior of the lipid material (Larsson,
1980). The most effective amylose-complexing emulsifiers have a high degree
of molecular freedom in an aqueous phase and exhibit lyotrophic mesomorphism. Micelles or liposomes of emulsifiers are the most effective polymorphs
for providing lipid monomers, and as a result, are more effective complex formers than lipids in other polymorphic states (Riisom et al., 1984; Eliasson,
1986a). Lysolecithin, a native lipid in cereal starches, complexes readily with

Carbohydrate/Emulsifier Interactions

85

amylose because it exists as micelles in solution with a high equilibrium concentration of monomers readily available for interaction (Takahashi and Seib,
1988). Crumb softeners are added to bread dough as J3-crystal monoglycerides
that convert into an a-crystalline state during baking that readily complex with
starch (Krog, 1981; Krog and Nybo-Jensen, 1970). Blends of emulsifiers that
form an emulsion in solution will be less effective starch-complexing agents
than pure monoglycerides.

4.5 Physical Properties of Starch/Emulsifier


Complexes
Physical characterization of starch/emulsifier complexes has provided information on the functionality of emulsifiers in starch systems. Techniques such
as differential scanning calorimetry, nuclear magnetic resonance, and electron
spin resonance spectroscopy have been especially useful.

4.5.1

X-Ray Diffraction Pattern

X-ray diffraction was one of the first methods used to identify and study inclusion complexes with starch (Mikus et al., 1946). X-ray diffraction provides information on the crystallinity of starches. A clathrate (inclusion) complex is
indicated when powder diffractograms of starch have a so-called V-type pattern.
X-ray diffraction has been used to confirm the presence of inclusion complexes
when an aqueous starch system has been heated in the presence of native lipids
or fatty adjuncts (Biliaderis et al., 1986; Deffenbaugh, 1990a; Eliasson, 1988;
Eliasson and Krog, 1985; Hanna and Lelievre, 1975;

Hoove~

and Hadziyev,

1981; Kugimiya et al., 1980; Osman et al., 1961; Rutschmann and Solms, 1990).
The helical conformation of amylose within the complex has also been shown using x-ray diffraction (Rutschmann and Solms, 1990). X-ray diffraction data does
suggest V-type complex specifically between amylopectin and lipid ligands
(Gudmundsson and Eliasson, 1990). It appeared important that "free" amylopectin was used, whereas amylopectin in waxy maize granules did not complex
(Eliasson and Ljunger, 1988; Evans, 1986; Kugimiya et al., 1980)
X-ray diffraction measurements have indicated that the cell dimension of
the helix is essentially the same for complexes with various emulsifiers containing one fatty acid moiety. Emulsifiers with two or more fatty acids are sterically excluded from the helix and complex formation is inhibited (Osman et
al., 1961). Most V complexes have a pitch of approximately 0.8 nm, indicating

86

Food Emulsifiers and Their Applications

that the coils of the helix are in contact with each other (French and Murphy,
1977). Jane and Robyt (1984) studied the lamella-like organization of V-type
complexes and concluded that starch chains were folded such that the chain
axes were perpendicular to the surface of the lamella.

4.5.2

Infrared Spectroscopy

Infrared spectroscopy has confirmed the presence of the carboxyl (Bates and
White 1986; Osman et al., 1961), methyl (Bates and White, 1986), or carbonyl
(Hahnel et al., 1995) group of complexing agent inside the starch helix in a
complex. A positive shift in the carbonyl IR absorption peak for GMS in a
starch complex has been measured (Hahnel et al., 1995). The strength of the
carbonyl group double bond increased because of the additional electron density of the starch complex.

4.5.3

Electron Spin Resonance

Stable free radical spin probes of fatty acids are used to study the interaction between the fatty acid and starch using electron spin resonance (ESR). Changes in
probe motion, reflected as changes of spectral line shape, occur when the probe
is adsorbed or in a highly viscous medium compared to measurements of the
probe in a solvent. The motion of a spin probe of stearic acid was greatly slowed
in the presence of wheat, high amylose maize, and waxy maize starches (Pearce
et al., 1985, 1987a). Binding was weaker in waxy maize than in the other
starches (Pearce et al., 1987b). The interactive moiety of the probe was shown to
be the fatty acid. Results were similar at room temperature or after heating to
95C and cooling back to room temperature. Binding was thought to occur
throughout the interior of the granule since surface absorption could not account
for the amount of probe bound. The presence of water-facilitated binding, presumably by enhancing solvent penetration into the granule (Nolan et al., 1986;
Pearce et al., 1985). Similar results were reported by Nolan et al. (1986) for
probe binding to maize starch at room temperature. Johnson et al. (1990) also
measured spin probe binding by regular and waxy maize starches at room temperature after heating to 95C. Heating was found to decrease the strength of the
complex formed after cooling, most likely because of altered molecular conformation. Spin probe penetration into the granule increased upon heating, due
both to increased granular surface area and increased accessibility of the interior
of the granule to the fatty acid probe.
Additional ESR analysis indicated that the fatty acid spin probe was slowly

Carbohydrate/Emulsifier Interactions

87

released during heating of starch to 95C as the complex dissociated upon


melting (Pearce et al., 1987a, b). Upon cooling, the probe was immobilized
again indicating reformation of the complex.

4.5.4

Nuclear Magnetic Resonance

Nuclear magnetic resonance (NMR) can measure changes in the environment


of food components on a molecular basis.

13

C NMR can detect changes in the

environment of carbon atoms in starch induced by complex formation with


emulsifiers (Deffenbaugh, 1990a; Jane et al., 1985).
Jane et al. (1985) reported a downfield shift in resonances of all starch carbons, but especially Cl and C4, in preformed inclusion complexes with amylodextrin. Deshielding of Cl and C4, the carbons in the glycosidic linkage, was
indicated by the downfield shift. Rotation about the CO bonds of the glycosidic
linkage must have occurred upon formation of a helix from a random coil. In solution state, l3C NMR, Cl and C4 of maize starch, experienced a downfield shift
in the presence of complexing agents during gelatinization between 55 to 75C
(Deffenbaugh, 1990a). No effect was seen > 70C in solution state. The complexes were forming during gelatinization but could not be detected in solution
state 13C NMR after precipitation of the starch/emulsifier complexes. Formation
of complexes with amylopectin in waxy maize starch was also detected.
Proton NMR has also been used to understand starch/ligand-complex formation. Signal intensity from amylose protons was reduced when Na palmitate was
added, an effect attributed to the loss of conformational mobility upon complex formation leading to extreme line broadening (Bulpin et al., 1982). The signal was restored upon heating to 90C, apparently due to dissociation of the thermally
reversible complex. Resonances of protons 3 and 5 of the glucosyl residues in cycloheptaamylose shifted upfield in the presence of lysolecithin (Kim and Hill,
1985). These changes indicated that these protons, directed toward the interior of
the molecule, were in a more hydrophobic environment upon inclusion of the
lysolecithin. No band shifts were found for amylopectin protons upon formation of
amylopectin/monoglyceride complexes (Batres and White, 1986).

4.5.5

Differential Scanning Calorimetry

Differential scanning calorimetry (DSC) data defines the temperature and enthalpy of thermal transitions. Gelatinization of starch is a water-mediated, endothermic melting phenomenon. Complexes between starches and emulsifiers
form exothermally (i.e., crystallize) during heating of starches with the emulsifier

88

Food Emulsifiers and Their Applications

and water (Kugimiya et al., 1980). The net enthalpy of starch gelatinization endotherm is reduced in the presence of external lipids due to simultaneous,
exothermic crystallization of starch/lipid complexes (Biliaderis et al., 1986;
Deffenbaugh, 1990a; Eliasson, 1983; Eliasson, 1986a; Eliasson and Ljunder,
1988; Eliasson et al., 1988; Evans, 1986; Kugimiya et al., 1980). Data is given
in Table 4.4 showing the effect of a sucrose ester emulsifier on gelatinization
temperature and enthalpy of various starches (Deffenbaugh, 1990a). Microscopy
and viscosity profile measurements, discussed previously, also detect a delay in
gelatinization of normal starches in the presence of emulsifiers. Depending on
moisture and emulsifier levels, DSC may not be sensitive enough to detect the
delay in starch gelatinization caused by emulsifiers (Deffenbaugh, 1990a; Kim
and Walker, 1992b). See Table 4.4.
Table 4.4 DSC parameters of starch gelatinization endotherm from
thermograms of starch with 0, I, 2 or 5% (starch wt basis) sucrose
ester emulsifier. (From Deffenbaugh, l990a.)

To

eq

Starch

0% SE

1% SE

2% SE

5% SE

Maize
Potato
Tapioca
Wheat
Waxy maize

66.661
59.741
63.54 1
58.701
69.03 1

66.53 1
59.831
63.97 1
59.101
68.40 1

66.491
59.751
64.03 I
58.42 1
68.401

66.42 1
59.611
63.90 1
59.021
68.13 1

Starch

0% SE

1% SE

2% SE

5% SE

Maize
Potato
Tapioca
Wheat
Waxy maize

72.83 1
64.751
70.191
63.691

72.591
64.75 1
70.64 1
63.72 1

72.69 1
64.891
70.82 1

72.661
64.60 1
70.31 1

74.751

74.171

63.301
74.291

63.671
74.24 1

Tpeq

MI (J/g)
Starch

0% SE

1% SE

2% SE

5% SE

Maize
Potato
Tapioca
Wheat
Waxy maize

13.441
16.931
18.191
10.61 1
16.901

11.5()2
16.641
15.282
9.58 12
17.011

10.612
16.2612
13.773
9.33 2
16.961

10.662
15.372
11.834
8.782
16.83 1

12 3 4

Different numbers indicate significant difference (P- 0.05) within starch type.

Carbohydrate/Emulsifier Interactions

89

Starch crystallites in most starches melt or gelatinize between 60 and 80C


in a first-order melting process. The insoluble starch/lipid complexes that form
during gelatinization typically melt between 100 to ll5C. At high moisture
levels, melting of the complexes is highly cooperative and only a single endotherm is observed.
The gelatinization endotherm is not observed in DSC rescans of starch samples because gelatinization is irreversible. Starch inclusion complexes melt
and recrystallize reversibly and, thus, occur in rescans. Identification of starch
complex-melting endotherms in first and subsequent DSC scans of starch samples is used to confirm the presence of starch/lipid complexes (Deffenbaugh,
1990a; Eliasson et al., 1988; Hoover and Hadziyev, 1981; Kugimiya et al.,
1980; Staeger et al., 1988; Szczodrak and Pomeranz, 1992).
DSC can be used to measure the relative thermal stabilities of starch inclusion complexes. The stability of the complex depends on the type of starch and
emulsifier, and is important because complex stability affects the rheological
changes during gelatinization (Eliasson, 1986b). Thermal stability (Tg) and
complex-melting enthalpy (All) decrease when a fatty acid chain is interrupted
with cis unsaturation (Eliasson and Krog, 1985; Stute and Konieczny-Janda,
1983; Raphaelides and Karkalas, 1988). Chain length of the lipid moiety has
no effect on

~and

may or may not affect Tg (Hoover and Hadziyev, 1981;

Eliasson and Krog, 1985; Raphaelides and Karkalas, 1988; Stute and
Konieczny-Janda, 1983). GMS forms a very stable complex with starch and
has some of the most profound effects on gelatinization properties.
Physical properties of starch/emulsifier complexes will depend on conditions
during crystallization. Multiple-melting endotherms for starch/emulsifier complexes or shifting of endotherms upon rescanning indicates the presence of different crystal polymorphs (Kugimiya et al., 1980; Biliaderis and Galloway, 1989;
Eliasson, 1988; Biliaderis et al., 1986; Bulpin et al., 1982; Paton, 1987). At the
onset of gelatinization, association of the amylose chain with a ligand provides
the conformational order (helicity) for nucleation that precedes crystallization.
Complexation during the first heating may be incomplete due to restricted mobility of amylose during gelatinization (Kugimiya and Donovan, 1981).
Small, imperfect crystals formed during gelatinization may have individual
helical segments distributed randomly without crystallographic register. An
annealing process occurs between Tg, amorphous glass transition temperature,
and Tm, crystallite melting temperature. Partial melting of less stable crystals
allows sufficient chain mobility for growth of more stable crystals. The melt-

90

Food Emulsifiers and Their Applications

ing/recrystallization process yields a more dense crystal composed of crystallites embedded in and molecularly continuous with disordered chain segments. Annealing of polymorphs as the temperature increases or during
rescans will increase crystallite melting temperature.
Different polymorphs of starch/emulsifier complexes may also exist within a
larger crystal that has folded onto itself (Eliasson, 1988). Complexes in folds or
at the surfaces of crystals would have a lower melting temperature than complexes further inside the crystal.

4.6

Summary

It is ironic that the interaction between emulsifiers and starches is "simply"


based on one phenomenon-the formation of inclusion complexes between
starch helices and fatty adjuncts. The amount of research devoted to understanding the interaction between starches and emulsifiers is daunting. Many
methods and measurements are used in these studies because each provides a
different perspective to understanding the extent and nature of the interaction.
The physical and sensory characteristics of many foods are largely defined by
starches. New and improved foods are developed because emulsifiers can be
used to modify the characteristics of starches in foods. Reduction of stickiness in
reconstituted potatoes is an example. The food industry is also dependent on
emulsifiers to improve shelf life of foods. The economic advantage of increasing
shelf life of bread with anti staling emulsifiers outweighs the cost of the emulsifier
by manyfold. Applications for starch-complexing emulsifiers will continue to expand as the properties of the interaction are better understood.

Acknowledgment
The author acknowledges with great appreciation the assistance of Mrs. Teresa
Booher in preparation of the manuscript.

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Chem., 24, 231-49.
- - - , Williams, C.B. (1944). Adsorption of fatty acid by the linear component of
corn starch, ]. Amer. Chem. Soc., 66, 1232-33.
Staeger, G., Escher, F., Solms, J. (1988). Formation of starch inclusion compounds during extrusion cooking, in Frontiers of Flavor (ed. G. Charalambous), Elsevier,
Amsterdam, pp. 639-54.
Strandine, E.F., et al. (1951 ). Effect of monoglycerides on starch, flour, and bread: a microscopic and chemical study, Cereal Chem., 28, 449-62.
Stute, V.R., Konieczny-Janda, G. (1983). DSC investigations of starches. Part II.
Investigations on starch-lipid-complexes, Starke, 35, 340-47.
Szczodrak, ]., Pomeranz, Y. (1992). Starch lipid interactions and formation of resistant
starch in high-amylose barley, Cereal Chem., 69(6), 626-32.
Takahashi, S., Seib, P.A. (1988). Paste and gel properties of prime corn and wheat
starches with and without native lipids, Cereal Chem., 65, 474-83.
Twillman, T.J., White, P.J. (1988). Influence of monoglycerides on the textural shelf life
and dough rheology of com tortillas, Cereal Chem., 65, 253-7.
Van Lonkhuysen, H., Blankestijn, J. (1974) Interaction of monoglycerides with
starches, Starke, 26, 337-42.
Zobel, H.F. (1984). Gelatinization of starch and mechanical properties of starch pastes,
in Starch: Chemistry and Technology, 2d ed. (eds. R.L. Whistler, J.N. BeMiller, and
E.F. Pachall), Academic, Orlando, FL, pp. 285-311.

FIVE

Protein/Emulsifier Interactions
Martin Bos, Tommy Nylander, Thomas Arnebrant,
and David C. Clark

5.1

Introduction

Many food emulsions are more complex than the traditional definition of an
emulsion: a colloidal dispersion of liquid droplets in another liquid phase.
This is mainly because the dispersed phase is often partially solidified or the
continuous phase may contain crystalline material, as in ice cream. However,
one characteristic that all emulsions have in common is that they are (thermodynamically) unstable. The four main mechanisms that can be identified in the
process of breaking down an emulsion are creaming, flocculation, coalescence,
and Ostwald ripening. There are two ways in which the process of breakdown
of an emulsion can be influenced. First, use of mechanical devices to control
the size of the dispersion droplets and second, the addition of stabilizing
chemical additives like low molecular weight emulsifiers and polymers to keep
it dispersed. The main purpose of the latter is to prevent the emulsion droplets
from fusing together (coalescence), often achieved by repulsive droplet/droplet
interactions. These interparticle interactions are determined mainly by the
droplet surface, which is coated with emulsifiers, often biologically surface-active components like proteins, mono- and diglycerides, fatty acids, or phospholipids. The forces most commonly observed are electrostatic double layer, van

95

96

Food Emulsifiers and Their Applications

der Waals, hydration, hydrophobic, and steric forces. They are responsible for
many emulsion properties including their stability.
The complex mechanisms involved in formation, stabilization, and destabilization of emulsions make fundamental studies on applied systems difficult.
One approach has therefore been to clarify the basic physical and chemical
properties of emulsions by the study of simpler model systems. The adsorption
behavior of single-emulsion components like proteins, fatty acids, surfactants,
or phospholipids at liquid/air or liquid/liquid interfaces have given information about surface activity, adsorbed amounts, kinetics, conformation, and surface rheology. The development of experimental techniques has made it
possible to extend these studies to multicomponent systems. This has provided
further information concerning competitive adsorption, displacement, and
complex formation, which can be related to emulsion and foam stability.
For further information concerning the physicochemical factors affecting
the emulsion structure as well as characterization of food emulsion stability,
the reader is referred to the reviews of Dickinson (1987a; 1988), Dickinson
and Stainsby (1982), and Tadros and Vincent (1983), and for the principles of
emulsion formation to the review of Walstra (1983) along with the other chapters in this book. In this chapter we will focus on the molecular interactions
between proteins and other surface-active components present at the interface
of the emulsion droplets that prevent the droplets from fusing together.
Understanding the interaction between these emulsifier components is the key
to increase the emulsion stability as well as to be able to tailor the structure of
these systems. Various surface-active components like lipids, low molecular
weight (LMW) surfactants, and even phospholipids will be regarded as emulsifiers. We will first discuss the stability of the protein in liquid solutions, which
is an important factor for their behavior in emulsion systems. Although the behavior at liquid/liquid and liquid/air interfaces can be best compared with the
situation in an emulsion or foam, we will also discuss some relevant studies
concerning the solid/liquid interface as well as the effect of emulsifiers on the
solution behavior of proteins. A number of techniques can be used to study
protein/emulsifier interactions, including surface film balance, ellipsometry,
Brewster angle microscopy (BAM), circular dichroism (CD), differential scanning calorimetry (DSC), surface rheology, fluorescence spectroscopy, and neutron reflectivity. It is beyond the scope of this chapter to discuss these
techniques in detail, but when necessary a brief explanation will be given.
The link between the molecular interactions between emulsifier compo-

Protein/Emulsifer Interactions

97

nents and the properties of food emulsions will be discussed in the last section
of this chapter.

5.2 Protein Stability


A protein molecule can interact with other molecules and surfaces in many
ways. This is mainly due to the complexity of the surface of the protein molecule, which comprises positive and negative charges, groups with hydrogenbonding capability, and nonpolar regions. Intermolecular and surface forces
can thus occur via electrostatic interactions, hydration forces, acid/base interactions, hydrogen bonding, and the van der Waals forces. The hydrophobic and
ionic interactions together with the gain in entropy due to conformational
changes are often regarded as the driving forces for protein adsorption (Norde,
1986; Haynes and Norde, 1994). The orientation of the protein may affect its
interfacial properties (Bos and Kleijn, 1995a, b). Proteins can be classified as
"soft" when they have low internal stability and consequently can change their
conformation upon adsorption, and "hard" if the protein has a high internal
stability (Arai and Norde, 1990a, b; Norde et al., 1991). However, from this
simplified description it is impossible to generalize the behavior of proteins at
interfaces, due to their difference in size, shape, and flexibility and their susceptibility to changes in solution conditions and properties of the interface.
One important aspect regarding emulsifying properties is the stability of a
protein in solution. It is important to distinguish the conformational stability of
the protein from protein aggregation or precipitation due to reduced solubility
at pH close to the isoelectric point, at high ionic strength, and/or through the
binding of ions or lipids. Note that aggregation or precipitation can occur without any conformational changes of the protein (Tanford, 1967), although the
two types of instability relate to each other. Information about conformational
stability of a protein can be obtained from circular dichroism measurements
(Townend, 1969), compressibility measurements (Gekko, 1986), and calorimetry (Privalov, 1979; 1988) and has been extensively reviewed by several authors (Privalov, 1979, 1982; Dill, 1990; Creigton, 1990).
Here, we will focus only on some important aspects. The amino acid sequence of the peptide chain determines its folding into ordered structural motifs (a helix, ~ sheets, etc.) that in turn associate to form tertiary and
quartenary structure. Introducing disulfide bridges will also increase the stability. As a result, the interior of globular proteins is very densely packed, with

98

Food Emulsifiers and Their Applications

a mean packing density of 0.74, which is comparable with the values found for
crystals and organic molecules (Richards, 1977). In this densely packed environment, short-range forces like van der Waals interactions and hydrogen
bonding play an important role (Privalov, 1978). In addition, the hydrophobic
interactions contribute significantly in stabilizing the structure, as first pointed
out by Kauzmann (1959). Accumulation of hydrophobic residues in the core of
the molecule leads to a strong folding force. In contrast, the polar groups prefer
the outside of the protein molecule due to their solvent affinity.
Due to the delicate balance of forces mentioned above, the protein is marginally stable (Pace, 1981; Privalov, 1988; Dill, 1990). Changing one amino acid
residue might stabilize or destabilize the protein (Matsumura et al., 1988).
Binding of a lipid, surfactant, or another protein molecule can also have the
same effect. For instance, it has been shown that at low surfactant-to-protein ratios, the binding can stabilize the protein against thermally induced unfolding,
as observed for the interaction between fatty acids or sodium dodecylsulfate
(SDS) and bovine serum albumin (Gumpen et al., 1979), as well as SDS and~
lactoglobulin (Hegg, 1980). On the other hand, increasing the surfactant-to-protein ratio above lO moles of SDS per mole serum albumin or 1 mole of SDS per
mole of J3-lactoglobulin monomer causes unfolding of the protein. Creamer
(1995) observed similar stabilization of ~-lactoglobulin against urea induced unfolding at 1:1 molar ratios between the protein and SDS as well as palmitate. In
many emulsions involving proteins and emulsifiers, the interaction could take
place at the interface. The proximity of an interface might disturb the stability
force balance of the protein, resulting in unfolding of the protein. The unfolding
of proteins upon adsorption is entropically favored (Dill, 1990; Norde, 1986) and
might be the driving force for adsorption where the contact between the hydrophobic domains and the aqueous environment are minimized by proper orientation of the molecule (Haynes and Norde, 1994).

5.3

Protein/Surfactant Interactions

5.3.1

Protein/Surfactant Interactions at Solid Surfaces

Protein/surfactant interactions of solid surfaces have been studied with the aim
of estimating the protein attachment strength to surfaces, for optimizing detergency processes, and for avoiding undesired adsorption in medical applications.
The major part of the work has been carried out with the purpose of characterizing the protein binding to the surface rather than the protein/surfactant interac-

Protein/Emulsifer Interactions

99

tion. Due to this fact, many studies referenced below are concerned with the degree of removal, or elution, of adsorbed protein by surfactant. We have, however,
included these data in this review and tried to evaluate them in connection with
the sparse data that allow understanding on a molecular level. Even if the data
mainly refer to solid surfaces, the basic principles are also valid at liquid interfaces such as those of the emulsion droplet. Since the process of surfactant interaction with proteins at interfaces is determined by the surfactant/protein, the
surfactant/surface and protein/surface interactions, the following brief introduction is intended to provide a background on surfactant association and adsorption, and on surfactant/protein interactions in solution.
The polarized structure of surfactants derives from the hydrophilic head
group and a hydrophobic, usually hydrocarbon, part that promotes interaction
with each other, other molecules, and surfaces by the hydrophilic head as well
as the hydrophobic moieties. These interactions can be expected to be of electrostatic or polar, steric, and hydrophobic type, and the total interaction is
often a sum of or a balance between two or more of these forces. As a consequence of the strong tendency for the hydrophobic chains to avoid contact with
water, self-association will take place both in solution and at interfaces (see
Section 5.4.4). The monomer concentration in solution will be strongly dependent on the association pattern and thus have a pronounced effect on the interfacial behavior (Lindman and Wennerstrom, 1980; Israelachvili, 1992).
The general features of surfactant adsorption are
l. At high surfactant concentrations [around the critical micelle concentra-

tion (erne)], the adsorption rate is fast.


2. For water-soluble surfactants, the adsorption is reversible upon dilution.
3. Usually a plateau in the adsorption isotherm is reached in the range of
the erne.
4. As a rule surfactants adsorb at hydrophobic surfaces. The amounts adsorbed are in the range of, or below, those corresponding to a monolayer.
5. In the absence of specific chemical interactions, ionic surfactants adsorb
only onto hydrophilic surfaces of opposite charge (Amebrant et al., 1989).
These aspects of surfactant adsorption are illustrated in Figure 5.1 for the
adsorption of dodecyl trimethyl ammonium bromide (DTAB) onto methylated
silica and in Figure 5.2, where a schematic adsorption isotherm is shown.
There is a vast literature concerning the association of surfactants at
solid/aqueous interfaces (Somasundaran and Kunjappu, 1989; Pashley and

100

Food Emulsifiers and Their Applications

0.05

"'su
'bn

0.04

0.03

6
0

~ooooosoo8~
~

<

0.02
"a
Q)

Ul

"a

0.01

<:t;

0.00

1800
Time (s)

_Q

3600

Figure 5.1 The adsorption of DTAB (2 X erne in phosphate-buffered saline pH 7, I=


0.17) onto methylated silica; rinsing took place after 1800 seconds of adsorption
(Wahlgren, 1992). (Reprinted with kind permission of the American Chemical Society,
Washington, DC)

iv

ii

c
Figure 5.2 A schematic illustration of an isotherm for adsorption of ionic surfactants
to hydrophilic surfaces (Somasundaran and Kunjappu, 1989). Different stages of the
isotherm are labeled (I) to (IV), and schematic drawings of possible structures that exist
in these regions are presented in Figure 5.3. (Reprinted with kind permission of the
American Chemical Society, Washington, DC.)

lsraelachvili, 1981; Scamehorn et al., 1982; Tiberg, 1994; Wangerud, 1994).


The structure of the surface aggregates at the plateau has been debated, and
surface micelles, finite bilayers, or infinite bilayers have been suggested for
hydrophilic surfaces. Indications of complete bilayers (Pashley and
lsraelachvili, 1981) or interpenetrating hydrocarbon chains (Somasundaran
and Kunjappu, 1989) have been found. Figure 5.3 is an illustration of possible

Protein/Emulsifer Interactions

101

association behavior of surfactants in the different regions found in the adsorption isotherm, as shown in Fig 5.2. The upper drawings correspond to a hydrophilic surface, the lower to a hydrophobic surface.
5.3.1.1

Interaction of Surfactants with Adsorbed Proteins. The removal of


preadsorbed proteins by surfactant has been extensively studied by Horbett and
coworkers (Bohnert and Horbett, 1986; Rapoza and Horbett, 1989, 1990b; Ertel

et al., 1991) in investigations into the adsorption strength of blood plasma proteins, particularly fibrinogen. They introduced the term "elutability" to describe
the degree of removal. The surfactant elutability of proteins is affected by factors
that are known to influence the binding strength of a protein to a surface. Thus,
surfactant elutability has been found to decrease with conditions favoring conformational changes, i. e., decreasing protein concentration (Rapoza and Horbett,
1989, 1990a), increasing temperature (Bohnert and Horbett, 1986; Rapoza and
Horbett, 1990a), time of adsorption or "residence time" (Rapoza and Horbett,
1989; Ertel et al., 1991), and decreasing stability of the protein (McGuire et al.,
1995a, b; Wahlgren et al., 1993). However, surfactant elutability will be influenced not only by protein properties but also by the type of surfactant (WelinKlintstrom et al., 1993; Wahlgren et al., 1993a, b; Wahlgren and Amebrant,
1991) and surface (Wahlgren et al., 1993, 1994; Elwing and Colander, 1990), as
further discussed next.

11-111

Figure 5.3

IVa

ivb

An illustration of probable arrangements of adsorbed surfactant molecules


at different degrees of surface coverage. Adsorption to hydrophilic surfaces (upper panels) and hydrophobic ones (lower panels). The illustrations are drawn to represent
structures having minimal water contact with the hydrophobic parts of the molecules.
The labels (I) to (IV) refer to structures that may exist in different regions of the
isotherm shown in Figure 5.2. The figures should be considered as schematic and other
structures, especially for ii to iii, have been suggested. (Reprinted with kind permission
of the American Chemical Society, Washington, DC.)

102

Food Emulsifiers and Their Applications

Influence of protein properties.

Even though ionic surfactants may inter-

act, more or less specifically with charged residues of proteins, especially so at


low concentrations (see Section 5.3.1.2), no clear relation could be established
regarding the influence of protein net charge on the interaction with ionic surfactants at high surfactant concentration (Wahlgren et al., 1993; Wahlgren and
Amebrant, 1991; McGuire et al., 1995c). This might, of course, be related to
the fact that in principle all proteins contain both negative and positive
charges except at extreme pH. In an effort to determine key protein parameters
for their interaction with surfactants, Wahlgren and coworkers studied the
DTAB-induced removal of six adsorbed proteins: cytochrome c, bovine serum
albumin, a-lactalbumin, ~-lactoglobulin, lysozyme, and ovalbumin from silica
and methylated silica surfaces (Wahlgren et al., 1993). For silica surfaces, it
was found that the removal of the proteins that were still adsorbed after rinsing
with buffer, increased with decreasing molecular weight, adiabatic compressibility [a measure of conformational stability (Gekko and Hasegawa, 1986)],
and increasing thermal denaturation temperature (Wahlgren et al., 1993). In
the case of hydrophobic (methylated silica) surfaces, differences between the
proteins were smaller. However, increasing molecular weight and shell hydrophobicity of the protein seemed to reduce the degree of removal. It was also
found that the removal did not relate to the degree of desorption of proteins
upon rinsing with buffer, indicating that the mechanisms for the two processes
are different. Recent experiments on stability mutants of bacteriophage T4
lysozyme show a convincing relationship between DTAB-mediated elutability
and the difference in free energy of thermal unfolding of the protein in comparison with the wild type (McGuire et al., 1995a). Thus it can be concluded that
factors relating to the structural stability of the protein are of major importance
and that an increased conformational stability increases the degree of elution.

Influence of surfactant properties.

Wahlgren and coworkers studied the

influence of different surfactant head groups on the desorption of adsorbed


lysozyme (Wahlgren, 1992; Wahlgren et al., 1993; Wahlgren and Amebrant,
1991) by surfactants at concentrations above the erne (an exception was triethylene glycol n-dodecyl ether [C 12 E3 , which does not form micelles (Mitchell et
al., 1986)]. The difference between the effect of sodium dodecylsulfate (SDS)
and cationic and nonionic surfactants on protein adsorption to hydrophilic surfaces was found to correlate to the strength of binding to protein in solution.
This suggests that above the critical association concentration (cac), complex

Protein/Emulsifer Interactions

103

formation between surfactant and protein is involved in the removal mechanism of proteins from hydrophilic surfaces. In the case of hydrophobic surfaces, the removal processes of protein by the different surfactants, including
non-micelle-forming ones, are in general more similar than for the hydrophilic
surfaces. This might be expected, due to the different orientation of the surfactant, and suggests a replacement mechanism, due to higher surface activity of
the surfactant (Wahlgren, 1992). Blomberg and coworkers used the interferometric surface force technique (lsraelachvili and Adams, 1978) to study the
interaction between lysozyme adsorbed on mica and SDSo (sodium dodecane
sulfonate) and SDS. They found that SDSo, which has a Krafft temperature
above room temperature and hence does not form micelles, had a minor effect
on the interaction between adsorbed lysozyme layers on mica (Tilton et al.,
1993), and from the small change in surface potential, they concluded that few
surfactant molecules were bound to the adsorbed protein. SDS showed a similar low binding to lysozyme on mica at low concentrations (up to 0.5 erne) but
caused a collective desorption of the protein at the erne of the surfactant, indicating that the cac to adsorbed lysozyme is in the range of its self-association
limit in solution (erne) (Blomberg, 1992). These studies show that anionic surfactants bind to an adsorbed layer of lysozyme, which is almost neutral after
binding of the positively net-charged protein to the negative mica surface. The
binding of surfactant thus leads to an increased negative charge of the layer,
which in the case of SDS finally leads to desorption of the protein. It is likely
that this is due to electrostatic repulsion between the negatively charged surface and the protein/surfactant complexes.
Nonionic surfactants are generally found to be ineffective in removing protein from hydrophilic surfaces (Wahlgren, 1992; Elwing et al., 1989). As mentioned above, these surfactants bind to a very low extent to protein in solution
(except when specific binding sites or pockets are present) and to the proteincovered surface. At hydrophobic surfaces, however, they usually have a considerable effect (Elwing et al., 1989; Wannerberger et al., 1994). This was
elegantly demonstrated in a study of surfactant interactions with proteins adsorbed at a surface with a gradient in wettability (Elwing et al., 1989).
The effect of chain length of alkyltrimethylammonium surfactants on the
elutability of fibrinogen at concentrations above their erne was found to be
small at both silica and methylated silica surfaces (Wahlgren, 1992). Rapoza
and Horbett (1988, 1990a) did not find any effects of chain length of sodium
alkyl sulfates on the elutability of fibrinogen and albumin down to a chain

104

Food Emulsifiers and Their Applications

length of 6 carbon atoms. However, they found, as expected, that the chain
length did influence the surfactant concentration at which the onset of protein
removal was initiated. The trend was similar to the one observed for the onset
of other cooperative binding events (e.g., micelle formation).
Rapoza and Horhett (1990h) found that surfactants with large head groups
such as Tween 20 gave rise to lower fibrinogen elutahility levels than other surfactants at polyethylene surfaces. Welin-Klintstrom et al. (1993) found that the
elutahility of fibrinogen adsorbed at surfaces with a wettahility gradient decreased with the bulkiness of the hydrophobic part of the surfactant. In this connection it was also found that nonionics showed an increased removal of
fibrinogen into the more hydrophilic region of the gradient surface when the
cloud point (phase separation temperature) was approached (Wahlgren et al.,
1995). These general observations of removal efficiency are in line with the findings of Backstrom and coworkers (Backstrom, 1987; Lindman et al., 1988), who
studied the removal of fat by different surfactants and found a maximum at conditions corresponding to an optimum in the packing of surfactant molecules at a
flat interface and those of Malmsten and Lindman (1989), who investigated,
among other variables, the effect of temperature on cleaning of hard surfaces.
Thus, it may be concluded that at high surfactant concentrations, head group
effects are, as expected, most pronounced at hydrophilic surfaces hut less important at hydrophobic ones. In addition, it appears that principles for detergency in
general, involving the packing efficiency of molecules at interfaces, are applicable to qualitatively describe the removal of proteins from the surface.

Influence of suiface properties. As previously described, the adsorption


and orientation of surfactants are dependent on the type of surface, and it is
therefore natural to expect that the way in which surfactants interact with proteins should be influenced by the characteristics of the surface as well.
Horhett and coworkers (Bohnert and Horhett, 1986; Rapoza and Horhett,
1989) studied the elutahility of fibrinogen and albumin at different polymeric
surfaces and found that the elutahility and the change in elutahility with time
differed between surfaces. These differences could not, however, he correlated
to surface energetics in terms of their critical surface tension of wetting.
Investigations into the elutahility of lysozyme and ~-lactoglobulin from methylated silica (hydrophobic) and oxides of silicon, chromium, and nickel by SDS
and cetyltrimethylammonium bromide (CTAB) showed that it did not follow
any rules relating to the charge of the surface. Instead, elutahility of ~-lac-

Protein/Emulsifer Interactions

105

toglobulin and lysozyme decreased roughly in the order silica > chromium oxide > nickel oxide (Wahlgren and Arnebrant, 1991). The similarity between
the behavior of the two oppositely charged surfactants indicates that the removability of protein in these cases mainly reflects the binding mode of the
protein to the surface.
Elwing et al. (1989, 1990) studied the surfactant elutability of proteins adsorbed to a surface containing a gradient in hydrophobicity and found large
differences in the amounts removed from the hydrophilic and hydrophobic
ends. In the case of a nonionic surfactant (Tween 20), the elutability was
largest at the midpoint of the gradient, which can be attributed to enhanced
conformational changes of the adsorbed protein at the hydrophobic end, in
combination with a lower efficiency of removal by nonionics at hydrophilic
surfaces. At hydrophobic surfaces the removal is generally high (Elwing et al.,
1989; Wannerberger et al., 1994). However, this may not be considered as evidence for weak binding of the proteins to the surface, but rather as an indication of the strong interaction between the surfactants and surface.

Simple models.

The observed effects of surfactants on adsorbed proteins

can be classified into two types of behavior, a short description of which follows (see Figure 5.4):

l. Complex formation between surfactant and protein that leads to desorption of protein from the surface. In this case the surfactant does not have
to adsorb at the surface, but it has to interact with the adsorbed protein.
2. Replacement of protein by surfactant. This implies that the interaction
between the surfactant and the surface has to be stronger than the interaction between the protein or the surfactant/protein complex and the
surface. Adsorption of the surfactant to the surface is essential in this
connection, but binding of surfactant to protein may not be required.
Of course, the surfactant might bind to the surface and/or to the adsorbed
protein without any net effect on the amount of protein adsorbed. A partial removal according to mechanisms (1) or (2) as illustrated in Figure 5.4 may be
the most common observation and has previously been suggested as one indication of the presence of multiple adsorption states of the protein (Horbett and
Brash, 1987).
It is important to note that surfactant can remove proteins without binding to
the surface [case (1), left side of Figure 5.4]. This could be described as a solubi-

106

Food Emulsifiers and Their Applications

Figure 5.4 An illustration of surfactant-protein interactions at solid interfaces. The top


drawing schematically shows an adsorbed layer of protein consisting of a removable
(type 1) and a nonremovable (type 2) fraction. The differences might be due to, e.g.,
conformational and/or orientational aspects. The bottom-left drawing illustrates formation of desorbing complexes of surfactant and protein (solubilization), and replacement
of protein by surfactant is shown to the right.

lization of the proteins by the surlactant. An interesting question is whether the


replacement of proteins by surlactant in case (2) (right side of Figure 5.4) is first
initiated by solubilization followed by surlactant adsorption. Nonionic surlactants interact to a very low extent with soluble proteins (Tanford, 1980), as for example can be concluded from the low amounts adsorbed to the adsorbed protein
layer (Wahlgren, 1992). These surlactants still remove proteins from hydrophobic surlaces, and it is thus evident that in this case it occurs through replacement of adsorbed protein molecules by surlactant.
5.3.1.2

Adsorption from Mixtures of Proteins and Surfactants.

Proteins and

surlactants can interact in solution to form surlactant/protein complexes that


may have different properties from those of the pure protein (Tanford, 1980;
Ananthapadmanabhan, 1993). Ionic surlactants are known to interact with
proteins in solution, and the interaction is generally stronger for SDS than for
cationic surlactants (Nozaki et al., 1974; Subramanian et al., 1986;
Ananthapadmanabhan, 1993). Nonionic surlactants are known to generally interact poorly with soluble proteins (Tanford, 1980) unless specific hydrophobic
binding sites exist. Three types of interactions are observed:
l. Binding of the surlactant by electrostatic or hydrophobic interactions to

specific sites in the protein, such as for ~-lactoglobulin (Tanford, 1980;

Protein/Emulsifer Interactions

107

Jones and Wilkinson, 1976; Coke et al., 1990: Frapin et al., 1990;
O'Neill and Kinsella, 1987; Kresheck et al., 1977; Clark et al., 1992;
Creamer, 1995), serum albumin (Tanford, 1980; Nozaki et al., 1974;
Brown, 1984; Ericsson and Hegg, 1985), and specific lipid-binding proteins such as puroindoline from wheat (Wilde et al., 1993).
2. Cooperative adsorption of the surfactant to the protein without gross conformational changes.
3. Cooperative binding to the protein followed by conformational changes
(Few et al., 1955; Subramanian et al., 1986; Su and Jirgensons, 1977;
Nelson, 1971).
These different interactions can occur in the same system upon increasing the
surfactant concentration. The conformational changes that occur in case (3)
can involve changes in secondary structure (Nozaki et al., 1974; Subramanian
et al., 1986; Su and Jirgensons, 1977). Several models for the protein/surfactant complexes have been suggested, e.g.: rigid rod (Reynolds and Tanford,
1970), pearl and necklace ( Shirahama et al., 1974), and flexible helix model
(Lundahl, 1986). In the cooperative region (2 and 3), above the critical association concentration (cac ), the interaction is mainly one of hydrophobic character (Tanford, 1980; Subramanian et al., 1977; Nelson, 1970).
The adsorption from surfactant/protein mixtures to hydrophobic solid surfaces is to some extent analogous to the adsorption at air/water or oiVwater interfaces, which have been the subject of frequent studies (Courthaudon et al.,
1991a, b; Clark et al., 1994). Competitive adsorption between proteins and
surfactants at these interfaces has recently been reviewed by Dickinson and
Woskett (1989). They conclude that, as expected, small surface-active components above a certain critical concentration will dominate over proteins at
these interfaces, since such components normally have higher surface activity
(superiority in lowering interfacial tension).
Experimentally it is observed that the presence of surfactants in protein solutions may influence the amount of proteins adsorbed to solid surfaces in
three different ways (Wahlgren 1992; Wahlgren and Arnebrant, 1991, 1992;
Wahlgren et al., 1993, 1995):

l. Complete hindrance of protein adsorption


2. Reduced amounts of adsorbed protein compared to adsorption from pure
protein solution
3. Increased amounts adsorbed

108

Food Emulsifiers and Their Applications

The complete lack of adsorption in case (1) could be explained by complex formation with a surfactant resulting in an entity that has no attraction to the surface
or direct/preferred surfactant adsorption, due to their higher surface activity and
diffusivity, unpending adsorption of proteins, or protein/surfactant complexes. In
cases (2) and (3), the formation of complexes in solution leads to a decrease or increase in the amounts of protein adsorbed, respectively. The presence of surfactant
influences the total amount of protein adsorbed by steric effects or changes in the
electrostatic interaction between complexes as opposed to native protein. In addition, the complex may adsorb in a different orientation than the pure protein, resulting in a positive or negative effect on the adsorbed amount.
Due to the different shapes of the adsorption isotherms of surfactants and proteins, the interaction with the interface is of course, as for protein mixtures,
strongly dependent on the relative concentration of the components. The special
character of the surfactant adsorption isotherms featuring the sharp increase in adsorbed amount in the range of their critical association concentration will influence
these events in a very pronounced way. Studies regarding these surfactant/protein
"Vroman effects" have been reported; for example, adsorption of fibrinogen from
mixtures containing Triton X-100 passes through a maximum (Slack and Horbett,
1988). The adsorption from ~-lactoglobulin/SDS mixtures at different degrees of
dilution was studied by Wahlgren and Amebrant (1992) (see Figure 5.5). At concentrations above the erne for the surfactant, the amount adsorbed corresponded to
a layer of pure surfactant and was found to increase after rinsing. At lower concentrations, the adsorbate prior to rinsing appeared to be a mixture of protein and surfactant, and the total amount adsorbed passes through a maximum. The
composition of the adsorbate after rinsing is most likely pure ~-lactoglobulin, since
interactions between the surface or protein and SDS are reversible.
At high degrees of dilution of the mixture, the absence of surfactant adsorption to the methylated silica, the nonreversible adsorption of ~-lactoglobulin,
and the observed partial desorption of the adsorbate from the mixture imply
that some SDS molecules are bound to ~-lactoglobulin molecules with a higher
affinity than to the surface (see Figure 5.5). The amount of protein adsorbed is
larger, even after rinsing, than for adsorption from pure ~-lactoglobulin solutions, and it can be concluded that SDS binding in this case facilitates the adsorption of protein.

Conclusions. Generally, it can be concluded that surfactants may interact with


interfaces through solubilization or replacement mechanisms, depending on sur-

Protein/Emulsifer Interactions

109

0.18

......-: --

..-...: .-- .

0.12

0.06

0
1000

100

Degree of dilution

10

Figure 5.5 The amounts adsorbed to a methylated silica surlace as a function of degree of dilution for a mixture of ~-lactoglobulin and SDS (0.2 w/w), in phosphate
buffered saline pH 7, I= 0 .17. The figure shows the adsorbed amount (!lg/cm2) after 30
minutes of adsorption (e) and 30 minutes after rinsing(+). In addition, the figure shows
the adsorption of pure ~-lactoglobulin, after 30 minutes of adsorption () and 30 minutes after rinsing (x). Finally, the adsorption isotherm of SDS is inserted (0). (From
Wahlgren and Arnebrant, 1992.)

factant surface interactions and surfactant/protein binding. Solubilization requires complex formation between protein and surfactant, and replacement requires adsorption of the surfactant to the surface. As for protein adsorption, one of
the most important protein properties affecting surfactant-induced desorption appears to be conformational stability. Differences between a competitive situation
and addition of surfactant after protein adsorption may derive from the alteration
in surface activity of protein/surfactant complexes formed in solution compared to
pure protein, the difference in diffusivity of surfactants and proteins affecting the
"race for the interface," and time-dependent conformational changes resulting in
"residence time" effects.
The effects observed at low surfactant concentration, below the erne, mvolve the specific binding of surfactant to some proteins and are not fully understood. Further, the exact prerequisites for solubilization versus replacement
as well as detailed information on molecular parameters such as aggregation
numbers are not known.

110

Food Emulsifiers and Their Applications

5.3.2 Protein/Surfactant Interactions at Liquid/Air


and Liquid/Liquid Interfaces
Interactions between proteins and surfactants at air/water and oil/water interfaces has attracted considerable study in recent years because the consequences of competitive adsorption of these two species at these interfaces can
often strongly influence dispersion (foam or emulsion) stability against coalescence. The majority of proteins have high affinity for interfaces, which they
saturate at comparatively low concentrations compared to low molecular
weight (LMW) surfactants (Dickinson and Woskett, 1989; Coke et al., 1990).
Thus, on a mole for mole basis at low concentrations, proteins reduce the surface tension to a greater extent than LMW surfactants. However, the opposite
effect is observed at high concentrations, because at saturation coverage with
LMW surfactants, the interfacial tension of the interface is usually lower than
that achieved by proteins, and as a result, the latter molecules will be displaced from the interface. The region where the two different components coexist in the interfacial layer is of greatest interest, since it is in this region that
the stability of the system to coalescence is most greatly affected. A clearer
understanding of this has emerged from direct study of the structures that separate the dispersed-phase of foams or emulsions, under conditions of high dispersed-phase volume (i.e., foam or emulsion thin films). Such structures form
rapidly in foams following limited drainage but may occur only in emulsions
after creaming of the dispersed phase.
5.3.2.1

Foam and Emulsion Film Stabilization.

Thin films are stabilized by

two distinct mechanisms; the one that dominates is dependent upon the molecular composition of the interface (Clark, 1995). Low molecular weight surfactants such as food emulsifiers or polar lipids congregate at the interface and
form a fluid-adsorbed layer at temperatures above their transition temperature
(see Figure 5.6a). When a surfactant-stabilized thin film is stretched, local
thinning can occur in the thin film. This is accompanied by the generation of a
surface-tension gradient across the locally thin region. Surface tension is highest at the thinnest point of the stretched film, due to the local decrease in the
surface concentration of emulsifier in the region of the stretch. Equilibrium
surface tension is restored by adsorption of surfactant from the interlamellar
liquid, which is of very limited volume in a drained thin film. This process is
called the "Gibbs effect." Alternatively, migration of the surfactant by lateral
diffusion in the adsorbed layer toward the region of highest surface tension

Protein/Emulsifer Interactions

111

may also occur (Clark et al. 1990a). Here, the surfactant drags interlamellar
liquid associated with the surfactant head group into the thin region of the film
and contributes to the restoration of equilibrium film thickness. This process is
often referred to as the "Marangoni effect" (Ewers and Sutherland, (1952).
In contrast, the adsorbed layer in protein-stabilized thin films is much
stiffer and often has viscoelastic properties (Castle et al., 1987). These derive
from the protein/protein interactions that form in the adsorbed layer (see
Figure 5.6b). These interactions result in the formation of a gel-like adsorbed
layer, recently referred to as a "protein-skin" (Prins et al., 1995), in which lateral diffusion of molecules in the adsorbed layer is inhibited (Clark et al.,
l990b). Multilayer formation can also occur and serves to further mechanically strengthen the adsorbed layer (Coke et al., 1990). When pure protein

(o) Syrfnctnru-alonc

(b) Pmtein-olpne

h igh mobility

Film sttetchiog

~
diffusion~

~Prot
~ cin

rapid

dcfonnntion

" " ' (c) Mixed mlem


rc:duc:ed mobility

/
no int.emct.i:ons

Pilm stretching
No Protein
defonnation

Slow diffusion

P'lm rui l r e.

Figure 5.6 Schematic diagram showing the possible mechanisms of thin-film stabilization. (a) The Marangoni mechanism in surfactant films; (b) the viscoelastic mechanism in protein-stabilized films; (c) instability in mixed component films. The thin films
are shown in cross section and the aqueous interlamellar phase is shaded. (Reprinted
with kind permission of the Royal Society of Chemistry, London.)

112

Food Emulsifiers and Their Applications

films are stretched, the change in interfacial area is dissipated across the film,
due to the cohesive nature of the adsorbed protein layer and possibly the deformability of the adsorbed protein molecules.
Thin-film instability can result in systems that contain mixtures of proteins
and low molecular weight surfactants (Coke et al. 1990; Clark et al., 1991a;
Sarker et al., 1995), as is the case in many foods. The origin of this instability
rests in the incompatibility of the two stabilization mechanisms: the Marangoni
mechanism relying on lateral diffusion, and the viscoelastic mechanism on immobilization of the protein molecules that constitute the adsorbed layer. One
can speculate that in a mixed system, competitive adsorption of low molecular
weight surfactant could weaken or interfere with the formation of protein/protein interactions in the adsorbed layer and destroy the integrity and viscoelastic properties of the adsorbed layer (see Figure 5.6c). This could be a
progressive process, with the presence of small quantities of adsorbed surfactant initially introducing faults or weaknesses in the protein film. Adsorption
of more surfactant could induce the formation of protein "islands" in the adsorbed layer. These structures could be capable of slow lateral diffusion but
would be too large to participate in Marangoni-type stabilization. Indeed, they
could impede surfactant migration in the adsorbed layer. Adsorption of progressively more surfactant would reduce the size of the protein aggregates still
further until the adsorbed protein was in its monomeric form. Ultimately, all
the protein would be displaced from the interface by the surfactant.
Two types of interaction are shown in the schematic diagram of the mixed system. First, there is an interactive process associated with the coadsorption or
competitive adsorption of the two different species at the interface. Second, many
of the functional proteins used in food production have physiological transport activities and therefore possess binding sites, which may allow the formation of
complexes with surfactants. Let us consider each of these processes in tum. The
transition in adsorbed-layer structure at the air/water and oil/water interface
caused by competitive adsorption between protein and emulsifiers has been studied in detail. Oscillatory surface shear (Kragel et al., 1995) and dilation (Clark et
al., 1993: Sarker et al., 1995) measurements have been carried out at the air/water interface and show an emulsifier-induced reduction in surface shear viscosity
and surface dilational elasticity at critical emulsifier/protein ratios. This is consistent with the cartoon depicted in Figure 5.6, where at a specific molar ratio the
emulsifier will break down protein/protein interactions in the adsorbed layer, resulting in a reduction in surface shear viscosity and dilational elasticity. Similar

Protein/Emulsifer Interactions

113

observations have been made at the oil/water interlace under conditions of constant shear in experiments where both components were present in solution when
the interlace was formed (Courthaudon et al., 1991) and when the competing surfactant was added subsequent to formation of the protein-adsorbed layer (Chen
and Dickinson, 1995). Direct measurements of changes in adsorbed-layer rheological properties in foam and emulsion films is possible using the fluorescence
recovery after photobleaching (FRAP) technique (Clark et al., 1990a; Clark,
1995). This method relies on the positioning of a fluorescent reporter molecule in
the adsorbed layer either by covalent labeling of the protein of interest with a fluorescent moiety such as fluorescein isothiocyanate (FITC) or by use of an amphipathic fluorescent molecule [e.g., 5-N-(octadecanoyl)amino fluorescein, ODAF] at
low (submicromolar) concentrations that are insufficient to alter adsorbed-layer
structure. FRAP allows direct measurement of the lateral diffusion coefficient of
the fluorophore in the adsorbed layer of the thin film. The presence of an extensive network of protein/protein interactions in the adsorbed layers arrests lateral
diffusion of the probe molecule, as is the case in all films stabilized by protein
alone (Clark et al., 1990b). This situation persists in the presence of emulsifier
until a critical ratio of emulsifier/protein is achieved. This is marked by the onset
of surlace lateral diffusion in the adsorbed layer. The technique serves as a useful
means of (i) comparing the resistance of different proteins to emulsifier-induced
adsorbed layer disruption, (ii) evaluating the effectiveness of protein modification
strategies at improving the resistance of proteins to competitive displacement,
and (iii) investigating the usefulness of natural food ingredients as crosslinkers of
proteins in the adsorbed layer.
5.3.2.2

Specific Binding of Proteins and Emulsifiers.

Let us now consider the

effects of specific binding of emulsifiers by food proteins on adsorbed-layer


properties at the air/water and oil/water interlace. There are many examples of
proteins that possess binding activity, including bovine serum albumin and ~
lactoglobulin (see Section 5.3.1.2). Investigation of the binding properties of
these proteins has been generally confined to studies in bulk solution. For example, the presence of a fluorescent tryptophan residue in the hydrophobic
cleft of ~-lactoglobulin (Papiz et al., 1986) has facilitated the study of emulsifier binding by fluorescence titration. Subsequent analysis of binding by conventional methods such as that of Scatchard (1949) allows determination of the
dissociation constant (Kd) of the complex formed. Typical examples of KJs for
~-lactoglobulin

are shown in Table 5.1. The effect of complex formation can

114

Food Emulsifiers and Their Applications

usually be detected by shifts in the surface-tension (y) curve (Dickinson and


Woskett, 1988). An example of this is shown for Tween 20 and ~-lactoglobulin
in Figure 5.7 (Coke et al., 1990). Surface-tension/concentration (y-c) curves
for Tween 20 alone and in the presence of a fixed concentration of ~-lactoglob
ulin (0.2 mglml; 10.9 J1M) are shown.
Table 5.1

Typical dissociation constants of emulsifier/P-lactoglobulin complexes


Emulsifier

Tween 20
L-a lysophosphatidylcholine, palmitoyl
Sucrose monolaurate
Sucrose monostearate
Sucrose monooleate
Sodium stearoyl
lactylate, pH 7.0
Sodium stearoyl
lactylate, pH 5.0
Lauric acid
Palmitic acid

Dissociation
constant

Reference

4.6 j.iM
166.J.!M

Wilde and Clark, 1993


Sarker et al., 1995

11.6 j.iM
1.02 j.iM
24.8 j.iM
0.26 j.iM

Clark et al., 1992


Clark et al., 1992
Clark et al., 1992
Clark, unpublished

0.30 j.iM

Clark, unpublished

0.7 j.iM
0.1 j.iM

Frapin et al., 1993


Frapin et al., 1993

The general features described earlier are evident with a comparatively low
concentration of protein causing a significant reduction in y. In the absence of
protein, yreduces gradually with increasing Tween 20 concentration. The gradient of the reduction in surface tension reduces at higher Tween 20 concentrations (> 30 J1M) but doesn't become completely flat due to failure to attain
equilibrium y, possibly due to the presence of a mixture of surface-active species
in the Tween-20 sample. In contrast, the curve in the presence of protein maintains a relatively steady surface-tension value of about 50 mN/m up to Tween-20
concentrations of 25 j.iM due to adsorption of the protein. This means that the
curve for the sample containing protein crosses that of Tween 20 alone. This is
strong evidence for complex formation between the two components, since the
curves cross due to a reduction in the concentration of free emulsifier in solution
due to that which interacts with the protein to form the complex.
Thus, great care must be taken when considering the surface properties of

Protein/Emulsifer Interactions

115

80

70

5
~

60

u.;0
~

Q)
~

Q)

<..>

50

$....
;::l
r:n.

40

30

20

40

60

80

100

Figure 5.7 Surface tension isotherm for Tween 20 in the absence (e) and presence
() of 0.2 mg/mL ~-lactoglobulin. The data were recorded after 20 minutes adsorption
and are therefore not at equilibrium.

solutions containing mixtures of interacting components. In the simplest case


of a single binding site, the two-component system becomes a three-component system comprising free emulsifier, free protein, and emulsifier/protein
complex. The relative proportions of the components present can be calculated
in the following manner (Clark et al., 1992). In the simplest case, the interaction of an emulsifier (E) with a protein (P) can be described by the expression
P+E~PE

(l)

where PE is the emulsifier/protein complex. Thus the dissociation constant

(Kd) for the complex can be expressed as

[P] [E]
[PE]

(2)

where the square brackets indicate molar concentrations of the different


species. It is also the case that

116

Food Emulsifiers and Their Applications

[P] = [P101]

[PE]

(3)

[E] = [E101]

[PE]

(4)

where [P101] and [E101] are the total protein and emulsifier m the system.
Substituting (3) and (4) in (2) gives

which can be solved for [PE] and can be used to calculate the relative concentrations of the three components. In addition, the binding data, which may
comprise a change in a parameter (e.g., intrinsic fluorescence) caused by formation of the complex may be fitted using this equation, provided there is a
single active binding site and the titration is carried out to saturation.
Alternatively, it is possible to determine the dissociation constant and number
of binding sites from the Scatchard (1949) equation

[E]

(6)

where v is the fraction of protein with occupied sites (i.e., [PE]/[P101]). If the
Scatchard plot of v against v/[E] gives a straight line, it indicates the presence
of only one class of binding site. The gradient of this line is -l!Kd, and the intercept on the xaxis gives the number of binding sites, n. If the Scatchard plot
does not give a straight line, then the shape of the curve obtained can be used
to identify if the observed binding is positively or negatively cooperative or the
presence of multiple independent sites. In the former case the Hill equation
can be used to determine the Kd and a cooperativity coefficient (Hill, 1910).
Evidence for the formation of a specific complex in solution by direct measurement or by crossovers in surface-tension/concentration isotherms is not
sufficient to allow the conclusion that intact complex adsorbs directly at the
air/water or oil/water interface. There are few studies that provide convincing
data that support such a hypothesis, and that which is provided often only indirectly points toward the presence of adsorbed complex at the interface. One
of the best understood systems is that of Tween 20 and ~-lactoglobulin (Coke

Protein/Emulsifer Interactions

117

et al, 1990; Wilde and Clark, 1993), which are known to interact in solution
to form a 1:1 complex characterized by a Kd = 4.6 ~' which has an increased hydrodynamic radius of 5. 7 nm compared to 3.5 nm for P-lactoglobulin alone (Clark et al., 1991b). Detailed measurements of the properties of
foam films formed from a constant concentration of 0.2 mg/mL mixed native
and fluorescein-labeled P-lactoglobulin as a function of increasing Tween-20
concentration (Wilde and Clark, 1993; Clark, 1995) have been reported. This
study revealed that between molar ratios (R) of Tween 20 to P-lactoglobulin of
0.2 to 0.9, there was a progressive increase in the thickness of the foam films
and a corresponding decrease in the amount of adsorbed protein to an intermediate level of approximately 50% of that which was originally adsorbed
(see Figure 5.8). These changes occurred prior to the onset of surface diffusion of the labeled protein as determined by the FRAP technique at R = 0.9
(Coke et al., 1990). The increase in foam-film thickness was unexpected since
protein displacement by the Tween 20 should have reduced the thickness of
the thin film. One persuasive interpretation of the data is that coadsorption or
trapping of the Tween-20/P-lactoglobulin complex in the adsorbed multilayers could account for adsorbed-layer thickening (Clark et al., 1994), since the
complex is known to have an increased hydrodynamic radius (Clark et al.,
1991b). Calculations reveal that this is a distinct possibility, since 16 to 49%
of the P-lactoglobulin present in solution will be in the complexed form in the
R-value range, 0.2 to 0.9. Further confirmation of this explanation comes from
measurements of foam-film thickness at different P-lactoglobulin concentrations but at constant R value. Film thickness data for P-lactoglobulin at 0.2
and 1.0 mg/mL are also shown in Figure 5.8. The completely different thicknesses observed at the two protein concentrations can be rationalized in terms
of the amount of Tween-20/P-lactoglobulin complex formed. The sharp reduction in film thickness observed in the 0.2 mg/mL P-lactoglobulin sample at R
= 0.9 occurs when there is 5.41-LM complex present in the sample. An equiv-

alent amount of complex is present in the 1 mg/mL sample at R = 0.1 and


could account for the immediate reduction in film thickness observed with the
1 mg/mL P-lactoglobulin sample and the complete absence of a film thickening step at low R values.
Further evidence supporting direct adsorption of the complex formed between

~-lactoglobulin

and Tween 20 comes from dynamic surface-tension

(Ydyn) measurements performed using the overflowing cylinder apparatus


(Clark et al., 1993). The Yclvn isotherm for Tween 20 showed classical sig-

118

Food Emulsifiers and Their Applications

40
0
0

--,._,

_..,

C l)

;::S
0

30

OJ

43
...
;::S

CIJ

-E
Cl)
Cl)

OJ

20

];a
u

~
4

Molar ratio (R)

Figure 5.8 A comparison of foam-film thickness and surface concentration data for ~
lactoglobulin samples as a function of Tween 20 concentration. Surface concentration
of FITC/~-lactoglobulin (0.2 mg/mL) as determined by fluorescence counts (.&); foamfilm thickness for samples containing 0.2 mg/mL () and l.O mg/mL (e) ~-lactoglobu
lin. R refers to the molar ratio of Tween 20: ~-lactoglobulin.

moidal behavior hut was shifted to increased surfactant concentration by approximately 2 orders of magnitude compared to the static measurements.
Inclusion of ~-lactoglobulin (0.4 mg/mL) in the initial solutions caused only a
small reduction in the measured

Ydyn

to 71 mN/m. This remained unaltered in

the presence of Tween 20 up to a concentration of 15 f.!M. Above this concentration a small hut significant further reduction in

Ydyn

was observed. The ef-

fect resulted in a small inflection in the Ydyn curve in the region corresponding
to 15 to 40 J.LM Tween 20. At higher Tween-20 concentrations, the curve for
the mixed system followed that of Tween 20 alone. The inflection in the

Ydyn

isotherm observed for the mixed system at concentrations of Tween 20 greater


than lO JlM could not he due to adsorption of Tween 20 alone since under the
prevailing conditions, the concentration of free Tween 20 was reduced by its
association with ~-lactoglobulin. Using Equation (5) it can he shown that the
Tween-20/~-lactoglohulin

complex is the dominant component in solution in

the Tween-20 concentration range of 15 to 35 flM (Clark et al., 1993). Indeed,


the

Ydyn

isotherm indicates that there is very little difference between the sur-

Protein/Emulsifer Interactions

119

face activity of the three components present in solution, Tween 20, ~-lac
toglobulin, and the complex, since

'Ydyn

behavior is dominated by the compo-

nent present in the highest concentration. Such observations provide


important evidence for direct adsorption of intact complex, and in this particular case provides evidence that the observed "induction period" in static y
isotherms could be due to the formation of protein islands (de Feijter and
Benjamins, 1987) and can be abolished if interactions between the adsorbed
protein molecules can be prevented, as is the case with the Tween-20/~-lac
toglobulin complex.
Direct adsorption of complex at the air/water interface also appears to have
importance in functional properties of certain lipid-binding proteins from
wheat called "puroindolines" (Blochet et al., 1991; Wilde et al., 1993). These
proteins show unusual behavior in the presence of lipids that they bind, in
that their foaming properties are generally unaltered and in some cases enhanced. A systematic study of the influence of interaction with lysophosphatidyl cholines (LPC) of different acyl chain lengths has been completed
(Husband et al., 1995) and has produced persuasive evidence of the importance of the complex on foaming activity. First, two isoforms of the protein
were investigated, puroindoline-a and -b (the b form has also been referred to
as "friabilin"). Puroindoline-b has a significantly increased Kd for LPC compared to puroindoline-a (i.e., 20-fold weaker binding) and the enhancement
of foaming properties is correspondingly reduced in the b form. Further stud-

ies of the binding of LPC to the a form revealed that the binding became
tighter with increasing acyl chain length, and higher concentrations of the
short-chain-length LPC are needed to achieve optimal foam stability enhancement. One interesting observation was that the micellar form of LPC
was the species that bound to the protein. This finding emerged from the observation that lauryl-LPC showed no interaction with the puroindoline-a until
the levels present exceeded the critical micelle concentration of 400

This indicates a cooperative binding since it takes place in this concentration


range, and any of the suggested structures for the protein/surfactant complexes described in Section 5.3.1.2, e.g., the pearl and necklace structure,
could be applicable. It seems increasingly likely that the functional properties of the puroindolines are linked to a role in the transport and spreading of
lipid at the air/water interface analogous to the role of amphipathic lung surfactant proteins such as SP-B, with which they have striking structural homology (Hawgood and Clements, 1990).

120

5.4

Food Emulsifiers and Their Applications

Protein/Phospholipid Interactions

Introduction. In this section we will discuss the interaction between phospholipids and proteins. Both proteins and phospholipids are major components
of biological membranes, and therefore it is not surprising that a vast number
of publications concerning protein/phospholipid interactions are related to understanding the biomembrane processes. This aspect has recently been reviewed in a book by Watts (1993).
In pharmaceuticals, phospholipids are often used as colloidal drug carriers
in drug delivery systems in various physical forms like liposomes or emulsions. These drug delivery systems are cleared from the circulation (bloodstream) by the reticuloendothelial system. It is believed that this clearance is
triggered by adsorption of serum proteins on the phospholipid interface and
will depend on the properties of both the protein and the phospholipid (Eidem
and Speiser, 1989; Tabata and lkada, 1990; Patel, 1992).
In untreated milk the fat globule is stabilized by a lipid membrane, composed
mainly of sphingo- and phospholipids. Homogenization of milk results in a total
area increase of the fat globules by some 6- to 10-fold, where the increased surface
is stabilized by proteins adsorbed from milk during the process. Phospholipids,
particularly phosphatidyl-choline Oecithin), are added to various processed foods,
where they act as emulsifiers alone or together with proteins. Therefore, proteins
and phospholipids, separately as well as in complexed form, contribute significantly to the physical properties of many systems of technological interest (e.g.,
emulsions and foams). The intention of this section is to emphasize the diverse nature of phospholipid/protein interaction and to point to some implications of this for
the physicochemical as well as the biological properties of phosholipidlprotein systems. It is also our intention to point at some of the major forces involved in phospholipid/protein interactions. We will highlight only some properties of importance
for the understanding of phospholipid/protein interactions. Therefore the classification of interactions made in this section, when the lipid is dispersed in solution,
at different interfaces and with lipid phases, is arbitrary. Assignment according to
polar/nonpolar, anionic/cationic, or soluble/insoluble phospholipids could also
have been possible.

5.4.1

Protein/Phospholipid Interactions in Dispersed Systems

Many proteins have the biological role of transporting molecules with hydrophobic properties, which are bound to a hydrophobic pocket in the protein. ~
Lactoglobulin is thought to transport retinol (Papiz, 1986; Sawyer, 1987; North,

Protein/Emulsifer Interactions

121

1989) but has also been shown to have high affinity for phospholipids, fatty
acids, and triglycerides (Diaz de Villegas, 1987; Creamer, 1995; Sarker et al.,
1995; Kristensen, 1995). An important application of lipid/protein interactions
was reported by Kurihara and Katsuragi (1993), who found that a lipid/protein
complex, formed between ~-lactoglobulin and phosphatidic acid, could mask
bitter taste. This property was suggested to be specific for phosphatidic acid
since no effect was observed for mixtures of ~-lactoglobulin and phosphatidylcholine, triglycerides, and diglycerides. Differential scanning calorimetry measurements confirm the presence of a specific interaction between phosphatidic
acid and ~-lactoglobulin since the presence of distearoylphosphatidic acid
(DSPA) as well as dipalmitoylphosphatidic acid (DPPA) thermally stabilized the
protein, which was not observed when the protein was mixed with phosphatidylcholine, phosphatidylethanolamine, or phosphatidylglycerol (Kristensen et al.,
1995). No interaction could be observed if the lipid contained unsaturated fatty
acid residues or if it was mixed in the gel state with the protein. Thus, the results
show that the interactions between ~-lactoglobulin and phospholipids are
strongly dependent on the acyl chain as well as the head group. This is not simply a question of having a negatively charged head group since no interaction
was observed for phosphatidylglycerol.
The influence of protein structure on lipid/protein interactions has been
demonstrated by Brown et al. (1983), who observed that native ~-lactoglobulin
is unable to bind to phosphatidylcholine vesicles. However, if the protein was
dissolved in a-helix-forming solvent, binding to the phospholipid was observed. Brown suggested that the acyl chains of the lipid interact with the hydrophobic interior of the a helix, while the polar head group is likely to
interact with the hydrophilic exterior of the protein (Brown, 1984). The partially unfolded proteins formed during food processing may give helix structures when interacting with the lipids and these lipid/protein complexes can
improve the emulsification process (Brown, 1984; de Wit, 1989).

5.4.2

Protein/Phospholipid Interactions at Solid Surfaces

A few studies have demonstrated the use of deposited phospholipid layers, deposited via the Langmuir-Blodgett technique or by spincoating, to follow the
interaction between proteins and phospholipids in situ by using ellipsometry
(Malmsten, 1994, 1995; Corselet al., 1986; Kop et al., 1984). This approach
can also be used to analyze the kinetics of interaction (Corsel et al., 1986; Kop
et al., 1984). The work of Malmsten (1994, 1995) showed that the interaction

122

Food Emulsifiers and Their Applications

of human serum albumin, lgG, and fibronectin from human plasma with phospholipids spin-coated onto methylated silica surfaces depends on the phospholipid head group. He found no interaction of proteins with phospholipids
that have no net charge or shielded charges like phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, and phosphatidylinositol, whereas interaction was observed with the phospholipid surfaces containing unprotected
charges like phosphatidic acid, diphosphatidylglycerol, and phosphatidylserine. No differences in adsorption behavior were found for spin-coated surfaces
and Langmuir-Blodgett-deposited phospholipid layers.
The consequences of nonspecific interactions with negatively charged dioleoylphosphatidylserine (DOPS) bilayzrs observed for fibrinogen and albumin
in contrast to the specific ones observed for prothrombin have been demonstrated by Corsel et al. (1986). By analyzing their data in terms of intrinsic
binding and transport rate, they found that the initial rate of adsorption of prothrombin was transport-limited, whereas the rate-determining step for the albumin interaction was the binding. The adsorption rate of fibrinogen was either
transport-limited or controlled by the binding, depending on pH and ionic
strength. Since the DOPS bilayer contained biological binding sites for prothrombin, the interaction was completely reversible. However, the interaction
of fibrinogen and albumin with the bilayer probably induced conformational
changes of the proteins and as a consequence the interaction was irreversible.
Kop et al. (1984) found that a high density of DOPS in the bilayer was needed
for the high-affinity binding of prothrombin. Consequently, an introduction of
dioleoylphosphatidylcholine (DOPC) into the bilayer markedly decreased the
affinity for binding.

5.4.3

Protein/Phospholipid Interactions at liquid/air Interfaces

Electrostatic interactions between protein molecules and lipid monolayers


have been shown to be important for phospholipid/protein interactions at the
liquid/air interface. A model food emulsion was used to study the interaction
between nitroxide homologs of fatty acids and milk proteins by following the
mobility of the nitroxide radicals using electron spin resonance (Aynie,
1992). It was found that at pH 7 the lipid/protein interaction was correlated
with the number of positive charges on the protein. Thus, the importance of
the interaction in the emulsions was found to decrease in the order <X51 -casein

> ~-lactoglobulin > ~-casein, suggesting that the interaction was of electrostatic nature. The work of Quinn and Dawson (1969a, b) concerning the inter-

Protein/Emulsifer Interactions

123

action between cytochrome c (positive net charge below pH 10) and phospholipids from egg yolk also suggest that it is determined by electrostatics. Their
results show that the limiting pressures for penetration are 20 and 24 mN/m
for phosphatidylcholine and phosphatidylethanolamine, respectively, whereas
penetration into the phosphatidic acid and diphosphatidylglycerol (cardiolipin) monolayers occurred up to pressures (< 40 mN/m) close to the collapse pressure of the film. Furthermore, the presence of sodium chloride
decreased the interaction. Later, Kozarac et al. (1988) confirmed the findings
of Quinn and Dawson with their reflection spectroscopy results. Cornell
(1982) observed a specific interaction between ~-lactoglobulin and egg yolk
phosphatidic acid (e-PA) in spread mixed films at low pH (1.3 and 4) where
~-lactoglobulin carries a positive net charge. No interaction was observed in
the neutral pH range or for egg yolk phosphatidyl choline, (e-PC). ~-lac
toglobulin, adsorbing from solution into a spread monolayers of palmitoyloleoylphosphatidylcholine (POPC) and palmitoyloleoylphosphatidylglycerol
(POPG), was found to interact with the lipids only in the acid pH range
(Cornell and Patterson, 1989). The highest amounts of ~-lactoglobulin
(Cornell and Patterson, 1989), a-lactalbumin, or BSA (Cornell et al., 1990)
bound to mixed monolayers of POPC and POPG were observed below the isoelectric point of the protein, when the lipid layer and the proteins carry an opposite net charge, whereas less was adsorbed around and almost nothing
above the isoelectric point. The interaction was also found to be reduced in
the presence of calcium as well as if sodium chloride was added (Cornell et
al., 1990). Bos and Nylander (1995) used the film balance to study the incorporaffon of ~-lactoglobulin into monolayers of distearoylphosphatidic acid
(DSPA), distearoylphosphatidylcholine (DSPC), and dipalmitoylphosphatidic
acid (DPPA) and some of their results are shown in Figure 5.9. The highest
rate of incorporation was observed for ~-lactoglobulin into the negatively
charged DSPA monolayer. The rate also increases with ionic strength of the
subphase, which probably is due to a decrease of the repulsion within the
phosphatidic acid protein monolayer. The incorporation into the zwitterionic
DSPC monolayers is, as expected, less salt-dependent. The importance of
electrostatic interactions for the incorporation of proteins and peptides in
lipid layers have also been reported for a number of non-food-related systems
like adsorption of SecA (Breukink et al., 1992), actin (Grimard et al., 1993),
IgG (Lu and Wei, 1993), and opioid peptides and opiate drugs (Bourhim et
al., 1993) into phospholipid monolayers.

124

Food Emulsifiers and Their Applications

DSPC

. I
''I'

, I
'I

DPPA

-3 [t
(j)

-3.5

~
-4

0_0

-4.5
0

10

15

20

25

30

ri (mN/m)

Figure 5.9 The rate of incorporation of P-lactoglobulin into monolayers of distearoylphosphatidic acid (DSPA), distearoylphosphatidylcholine {DSPC), and dipalmitoylphosphatidic acid (DPPA) versus surface pressure (II). The data were recorded at
constant surface pressure by measuring the area increase of the lipid monolayer
spread on a protein solution containing 1.15 mg/L in 10 11M phosphate buffer pH 7,
with 0 11M (e), 50 11M (), or 150 11M (A) sodium chloride. The rate in mg/m 2 was
calculated from the area increase using the IT-area isotherm of spread monolayers of
P-lactoglobulin. Data adopted from Bos and Nylander (1995), where the experimental
details are also given.

Besides electrostatic interactions, hydrophobic interactions (based on the


hydrophobic effect (Tanford, 1980) between protein and phospholipid are also
important. Para-K-casein, a product of the cleavage of K-casein, retains its
ability to interact with DMPC monolayers due to its more hydrophobic nature,
whereas the macropeptide does not (Griffin et al., 1984). The adsorption of~-

Protein/Emulsifer Interactions

125

lactoglobulin into phosphatidic acid monolayers (Bos and Nylander, 1995)


showed that incorporation was faster into a distearoyl monolayer than into a dipalmitoyl monolayer, especially at higher surface pressures (see Figure 5.9).
This shows that the incorporation also is dependent on hydrophobic interactions. Bougis et al. (1981) showed that for the incorporation of snake venom
cardiotoxins into lipid interfaces, not only electrostatic interactions but also
hydrophobic interactions are important.
Structure of Protein/lipid Monolayers. The structure of mixed protein/
phospholipid films depends on both the properties of the components and their
interaction. It has been shown that conformational changes of the proteins during
or after the adsorption at the liquid/air and liquid/liquid interfaces influence the

5.4.3.1

properties of the protein film (Graham and Philips, 1979; Mitchell, 1986;
Dickinson and Stainsby, 1982). Circular dichroism spectra of ~-lactoglobulin
bound to phospholipid monolayers were found to be similar to those recorded in
solution, indicating that the conformation of the protein did not change significantly when interacting with the lipid monolayer (Cornell and Patterson, 1989;
Cornell et al., 1990). Katona et al. (1978) showed that the~ form of a hydrophobic
myelin protein was more able to interact with sphingomyelin than the <X form, indicating that the phospholipid is able to recognize different conformations of the
protein. Furthermore, the physical state of the phospholipid phase is important for
the interaction between protein and lipid. Fidelio et al. (1986) reported a more favorable interaction between melittin and phospholipids when the lipid film was in
a more liquid expanded state. This might be related to the fact that the protein is
able to penetrate into the phospholipid layer (more hydrophobic interactions).
Interaction (adsorption) of proteins with lipids might also occur at higher surface
pressures, where the lipids are in the condensed or crystalline phase. The protein
then is not able to intercalate the phospholipid monolayer, but might bind to it
forming a second layer, like actin does on negatively charged liposomes through
electrostatic interactions (Grimard et al., 1993).
Another interesting point for discussion is the homogeneity of the mixed
phospholipid/protein film. The homogeneity of mixed protein/lipid films has
been studied by, for instance, electron microscopy of films transferred to a solid
support by Cornell and Carroll (1985). They found that only lipids with the
chains in liquid state, e-PA, dioleoylphosphatidylcholine, and dioleoylphosphatidylethanolamine, formed homogenous films with ~-lactoglobulin, whereas
DPPA and DSPC formed heterogeneous layers. Cornell and Patterson (1989) and
Cornell et al. (1990) observed penetration of ~-lactoglobulin, a-lactalbumin, or

126

Food Emulsifiers and Their Applications

BSA into mixed monolayers of POPC and POPG at such high lipid pressure that
they found it unlikely that the proteins could penetrate into a protein layer of
their own at this pressure. Thus, they concluded that the formation of pure protein patches is unlikely and that portions of the protein is suggested to be intercalated in the lipid monolayer. Fluorescence microscopy together with the
surface-film balance technique has also been used to study the structure of
mixed phospholipid cytochrome c and b films (Heckl et al., 1987). It was found
that proteins were located mainly in the fluid membrane phase that coexisted
with solid lipid domains without protein. The penetration in the lipid monolayer
was reduced with increasing pressure. Cytochrome c (positively charged) was
found to interact with dimyristoylphosphatidic acid (DMPA) mono layers but not
with dipalmitoylphosphatidylcholine (DPPC) layers, from which it was concluded that the interaction was largely of electrostatic nature. The effect of different proteins on the organization of lipid monolayer has been demonstrated by
Aynie et al. (1992). Their results indicate that the IX8ccasein in contrast to ~-lac
toglobulin and ~-casein, probably due to the stronger lipid/protein interaction,
induce an ordering of a monolayer of nitroxide fatty acids on the surface of an
emulsion droplet.

5.4.4

Effect of Lipid Phase Behavior on Protein/Lipid Interactions

As discussed previously, the interaction between surfactants and proteins usually takes place via monomers in the specific binding regime, whereas the cooperative association usually takes place in the range of the erne. However, in
the case of lipids with low aqueous solubility, the association structures are
generally already present when the lipids are mixed with the protein. Thus, the
formation of these structures has a profound impact on protein/lipid interactions. It is therefore important to be familiar with the phase behavior of the
particular lipid under investigation and often seemingly conflicting results can
be derived from differences in the phase structure of the lipids. For instance,
we have observed that the interaction between ~-lactoglobulin and phosphatidic acid occurred only when the lipids were present as a dispersion, but not
when they were mixed in the gel state with the protein (Kristensen et al.,
1995). Hence, we will briefly discuss lipid phase behavior, although a more
detailed description is given elsewhere (see Larsson, 1994).

5.4.4.1

Polar Lipid Phase Behavior. The polar lipids, having large charged or
uncharged polar groups, have an amphiphilic nature and will thus associate in

Protein/Emulsifer Interactions

127

aqueous systems. The common feature for this self-association is the formation
of a polar interface, which separates the hydrocarbon and water regions. An
overview of some of the main types of structures is given in Figure 5.10. The
hydrocarbon chains can exist either in a fluid state, as in liquid crystalline
phases, or in a solid state, as in the lipid gel phases (Larsson, 1994).
Polar lipids can be further divided into two classes on the basis of their interaction with water:

l. Lipids and synthetic analogs that are water-soluble in monomeric and


micellar form, i.e., surfactants.
2. Lipids with very low water solubility, but with the ability to swell into liquid crystalline phases.
The water-soluble polar lipids (e.g., ionized fatty acids, bile salts, and synthetic surfactants, charged or uncharged) have monomeric solubility in the millimolar range and form micelles at higher concentrations. The critical micelle
concentration (erne) is considered to be a narrow concentration range, within
which aggregates start to form by a strong cooperative process (Lindman and
Wennerstrom, 1980). The driving force for micelle formation is the hydrophobic
interaction (see Tanford, 1980). For ionic amphiphiles, erne also depends on the
ionic strength, since addition of salt reduces the electrostatic repulsion between
the charged head groups. At low water content an inverse micellar structure, the
1 2 phase, is formed in which the hydrocarbon chains form the continuous
medium and the aqueous medium is present within the micelles.
A common feature of the two classes of polar lipids is the tendency to form
lyotropic liquid crystalline phases (see Figure 5.10). The main features of the
most commonly found mesophases were determined by Luzatti and coworkers
in the early 1960s by x-ray diffraction [reviewed by Luzatti in 1968 (Luzatti,
1968)]. Later spectroscopy studies have given a considerable number of contributions to the understanding of the dynamic nature of these phases. The
lamellar phase (La) consists of stacked infinite lipid bilayers separated by water layers, while the hexagonal phases consist of infinite cylinders, having either a hydrocarbon core (H 1) or a water core (Hu) The cubic phases have been
shown to exist in a number of lipid systems (Fontell, 1990) and their possible
role in biological systems has been discussed by Larsson (1989). Since the cubic phases are isotropic and highly viscoelastic, they can be hard to analyze
and to handle. Different structures of the cubic phases have been suggested,
which depend on the particular lipid system (Larsson, 1989; Lindblom and

128

Food Emulsifiers and Their Applications

Reversed micelles
(L2)
(Solid)
(Melt)

<-Cubic

Reversed hexagonal
(HII)

i=
<1::

a:
f-

w
u

"Water-in-oil"
<-Cubic (Q)

f-

<1::
f-

Lamellar
(La)

I"Mirror plane" I

<1::

LL.

a:

::l
(/)

"Oil-in-water"

<-Cubic

(/)

<1::

w
a:
u

Hexagonal
(Hi)

<Cubic

(H20)

Micelles
(L1)

Figure 5.10 Commonly formed association structures by polar lipids. Phase transitions can be induced by changes in water content, temperature, or interaction with
other solution components, like proteins. The lamellar liquid crystalline phase (LJ ,
can be regarded as the mirror plane, where the aggregates are of the "oil-in-water" type
on the water-rich side and of the "water-in-oil" type on the water-poor side (Fontell,
1992). On both the water-rich and water-poor sides of the L" there are two possible locations for cubic phases. Other "intermediate phases" may also occur.

Rilfors, 1989; Fontell, 1992). One type of cubic structure, consisting of rodlike aggregates, has been proposed by Luzatti et al. (1968). The two polar rodlike networks formed in this way give two continuous and unconnected
systems of fluid hydrocarbon chains. The other main type of cubic phases, observed for water-insoluble lipids, are bicontinuous and based on curved nonintersecting lipid bilayers, which are organized to form two unconnected
continuous systems of water channels (see Lindblom et al., 1979; Larsson,
1989). If an interface is placed in the gap between the methyl end groups of

Protein!Emulsifer Interactions

129

the lipid, it will form a plane that can be described as a minimal smface
(Andersson et al., 1988; Larsson, 1989). A minimal surface has zero average
curvature at any point on the surface; that is, in all points the surface is as concave as it is convex. A minimal surface exhibiting periodicity, as in the cubic
lipid aqueous phase, is termed an "infinite periodic minimal surface" (IPMS),
and three types of IPMS, with different geometry, have been shown to be important in lipid systems (Andersson et al., 1988; Larsson, 1989).
With decreasing water content, the phase behavior of the polar lipids often
follows this sequence: hexagonal phase (HJ ~ lamellar phase (LJ for watersoluble lipids and lamellar phase (LJ ~ reversed hexagonal phase (H1J for
lipids with low water solubility. Cubic liquid crystalline phases (Q) often occur
in between these. Phase transitions can also occur with changes in temperature; with increasing temperature the sequence of thermal transitions is usually the same as with decreased water content.
In nature and in many technical applications the aggregates consist of a
mixture of different lipids, which either exist in a homogenous mixture or separate into domains. As discussed in the review by Raudino (1995), the lateral
distribution in these mixed aggregates is influenced by a number of factors like
ionic strength, presence of polymers/proteins, as well as the composition of the
lipids; thus it is hard to give any general rules to predict when phase separation will occur.
Some Aspects of the Interaction between Proteins and Lipid Structures.
Interactions between proteins and lipid structures are of fundamental importance
in biological membranes, as well as in food emulsions. Our aim is to give some ex-

5.4.4.2

amples of the effects the interaction can have on the lipid phase behavior as well
as on the protein structure. The same type of molecular forces, as discussed in
previous sections, are responsible for the interaction.
Much of the work on lipid phase transitions induced by proteins and peptides has been focused on intrinsic and peripheral membrane proteins. Many
of these proteins carry a positive net charge at neutral pH, and most biological
membranes are neutral, zwitterionic, or negative under physiological conditions (Sankaram and Marsh, 1993). Binding of these proteins generally takes
place via direct electrostatic interaction between basic residues, like lysine
and arginine, and the opposite charges on the lipids. However, other effects
like hydrophobic interaction and steric factors can contribute significantly.
Gramicidin, an uncharged and hydrophobic pentadecapeptide, favors the tran-

130

Food Emulsifiers and Their Applications

sition lamellar phase --treversed hexagonal phase (Hu) in phosphatidylcholine


and phosphatidylethanolamine systems (Chupin et al., 1987). It was suggested
that the ability of the peptide to promote cylindrical (curved) structures, like
the hexagonal phase, is due to its conical shape and strong intermolecular attractive forces. An example of protein/lipid interactions that promote the reverse transition has been presented by Fraser et al. (1989). They found that
myelin basic protein could, due to its flexible structure and proper charge distribution, interact through electrostatic as well as nonpolar interactions with
PE and mixed PE/PS hexagonal phase and thereby promote the formation of
lamellar phase. It was proposed that the properties of myelin basic protein in
stabilizing the lamellar structure could be related to the stability of myelin
sheath multilayers (Fraser et al., 1989). Heimburg et al. (1991) reported that
binding of cytochrome c did not promote inverted hexagonal phase of DMPG
alone or in mixtures with neutral lipids. On the other hand, the recorded data
suggested the formation of an isotropic phase, with increased curvature of the
lipid layer. Our recent work on the interaction between cytochrome

and

monoolein in the cubic phase shows that the transition of cubic to inverted
hexagonal phase occurred at a much lower temperature at high protein content
than without protein being present (Razumas et al., 1995).
The NMR studies by Spooner and Watts (1991a, b) show that the interaction between cardiolipin bilayers and cytochrome c appears to cause extensive
unfolding of the protein. Similar observations have been made by Heimburg et
al. (1991), who reported that cytochrome c binds to bilayers of phosphatidylglycerol and mixtures between neutral and anionic lipids in two forms:

l. Close to the native conformation in solution


2. Unfolded with the heme crevice opened.

It was observed that in the fluid state of pure DMPG and DOPG, the bound
protein existed in the more unfolded form (II), whereas in the gel state of
DMPG the native form (I) prevailed. When DOPG, was mixed with nonionic
(DOG) or zwitterionic (DOPC), the bound fraction of the native form (I) increased with the content of neutral lipids.
The work of Minami et al. (1995), in which incorporation of P-lactoglobulin
and P-lactalbumin with negative net charges and lysozyme with positive net
charge at neutral pH, into a sphingomyelin/palmitate gel phase, was studied
found no correlation with the protein net charge. Instead, the amount of protein, which could be dissolved in the thin aqueous layer of the gel phase was

Protein/Emulsifer Interactions

131

suggested to be limited by the dimension of the layer. This is likely to be reduced as a consequence of the osmotic stress exerted by the "outside" solution
phase at high enough protein concentration (Minami et al., 1995).
The lateral phase separation of vesicle bilayers, containing a mixture of
phosphatidic acid and phosphatidylcholine, in the presence of lysozyme has
been followed by differential scanning calorimetry (Raudino and Castelli,
1992). The protein/lipid interaction, which was shown to be dependent on
electrostatic forces, thermally stabilized the protein. As discussed by Raudino,
the lateral phase separation occurring when protein binds to heterogeneous hilayers can eventually lead to the formation of defective lamellar phases, socalled ribbon phases, where the broken lamellae are limited by curved rims
(Raudino, 1995).
It has been shown that a number of proteins can be entrapped in the cubic
monoolein/water phase (Ericsson et al., 1983; Razumas et al., 1994; Razumas
et al., 1995). Not only can enzymes up to 590 kD be entrapped in the cubic
phase but also their activity is retained for a longer time compared to when
they are in bulk solution (Razumas et al., 1994).
How will the structure of the cubic phase be affected by the entrapment of a
protein? It seems that properties like viscoelasticity, microscopic appearance,
and the nature of x-ray diffraction pattern are basically the same as for a pure
monoolein/water phase (Ericsson et al., 1983; Portmann et al., 1991; Landau
and Luisi, 1993; Razumas et al., 1994; Razumas et al., 1995). However, there
are some important differences, indicating that the amount of protein as well
as the amount of water determines the phase behavior (Ericsson, 1986). The
study of Razumas et al. (1995) of the cubic monoolein/cytochrome c aqueous
phase also confirms this and points to the influence of electrolyte present.
Differential scanning calorimetry and enzyme activity measurements show that
lysozyme retains its conformation and activity in the cubic phase (Ericsson et
al., 1983). Similarly, it has been observed by circular dichroism measurements
that bacteriorhodopsin and melittin (Landau and Luisi, 1993) as well as achymokypsin (Portmann et al., 1991) retain their native conformation. On the
other hand, differential calorimetry data, and other observations on cytochrome c entrapped in the cubic phase, suggested some interactions that affect the thermal stability of the protein as well as of the cubic phase (Razumas
et al., 1995). Razumas et al. (1994) used a cubic monoolein-aqueous-phasecontaining enzyme as the biocatalytic layer in amperometric and potentiometric biosensors. They observed large differences in the stability, which

132

Food Emulsifiers and Their Applications

decreased in the order lactate oxidase > creatinine deiminase > glucose oxidase > urease, and corresponded to the order of increasing molecular weight.
Further investigations have shown that the introduction oflecithin in the cubic
phase can increase the stability of entrapped glucose oxidase (Nylander et al.,
1995). The mobility of the entrapped enzyme is limited in the cubic phase
compared to bulk solution (1995), whereas the diffusion measurements of
Mattisson et al. (1995) show that a typical substrate molecule, like glucose,
can move relatively freely into the cubic phase. These studies point to an important potential application of cubic lipid phases as a flexible and biocompatible matrix for the immobilization of enzymes for synthesis as well as for
analytical applications. The cubic monoglyceride phases also have the ability
to solubilize lipophilic proteins as well as relatively large amounts of membrane lipids. Larsson and Lindblom (1982) have reported the solubilization of
a-gliadin, which is a hydrophobic wheat protein fraction, into a cubic 1monoolein aqueous phase, with the same characteristics as a binary cubic 1monoolein phase. In this case, the proteins most probably are dispersed in the
lipid bilayer region of the cubic phase.

5.5. Protein/Emulsifier InteractionsFood Applications


In the previous sections we have treated various aspects of intermolecular
forces that have to be considered for the understanding of protein/emulsifier
interactions. Most of the results that have been presented have been obtained
from well-defined model systems. It should be born in mind that in an applied
system, we often have to consider one or several types of molecular interactions, that work either synergistically or antagonistically. We will present some
observations where the versatile nature of protein/emulsifier interactions are
demonstrated. In the last section we will give a brief summary of the principles
given in this chapter.

5.5.1

Lipolytic Enzymes and Protein/Lipid Interactions

The action of lipolytic enzymes is of importance in a number of food applications or related areas, ranging from their use in detergents, as tools in modifying lipids for the breakdown of acylglycerides both as unwanted side effects, to
the naturally occurring process in the human intestine. It is well known that lipases work mainly at an interface and thus they are an important example of

Protein/Emulsifer Interactions

133

lipid/protein interactions at interfaces. Excellent reviews regarding lipase action have been written by Verger and Pattus (1982) and Pieroni et al. (1990),
but we will highlight only some aspects in relation to the mechanisms of protein/lipid interactions. There are several types of lipases that act on phospholipids and triglycerides, but we will mainly discuss lipases catalyzing the
hydrolysis of the ester bonds of triacylglycerols. The enzymatic activity is determined by the concentration of lipolytic enzymes associated with the lipid
film and can be inhibited by various proteins (Gargouri et al., 1984).
Experiments carried out with mixed protein/dicaprin films transferred over
pure buffer yielded evidence that inhibition of hydrolysis was caused by proteins bound to the dicaprin film rather than by a direct interaction between
protein and lipase in the bulk phase (Gargouri et al., 1985). Furthermore,
since some lipases were inhibited by adsorption of proteins at the lipid layer,
whereas other lipases were still able to hydrolyze a mixed protein/phospholipid layer, indicating that the inhibition of some lipases cannot be attributed
merely to steric effects hindering accessibility to dicaprin molecules within
the film. Surface concentration measurements of inhibitory proteins showed
that only 5 to 9% of the area of a mixed lipid/protein film was covered by inhibitory proteins, implying that long-range electrostatic forces are likely to be
involved in the inhibition as well as parameters such as surface viscosity and
surface potential. However, similar inhibitory effects caused by melittin (pi >
10) and 13-Iactoglobulin A (pi = 5.2) at pH 8.0 strongly suggest that the nature

of the inhibition is not an electrostatic phenomenon, but might be assigned to


the effect on the properties of the hydrocarbon moiety of the lipid (Gargouri et
al., 1987, 1989; Pieroni et al. 1990). This is supported by the finding that there
is a correlation between inhibition of lipase activity and the ability of the inhibitory protein to penetrate into the phospholipid monolayer.
The necessary interface can also be provided by a particular lipid structure,
e.g., the cubic lipid aqueous phase. In a simple experiment, Wallin and
Arnebrant (1994) demonstrated that a cubic phase was decomposed much
faster by the action of lipase from Humicola lanuginosa than the oil phase in a
reference sample consisting of triolein and aqueous phase. This was attributed
to the much larger interfacial area of the cubic phase. In an in vitro study of
lipolysis of triglycerides in a intestinal-like environment, Patton and Carey
(I 979) observed, along with the initial crystalline phase, a viscous isotropic
phase composed of monoglycerides and fatty acids, which is identical to the
one formed in monoglyceride systems.

134

Food Emulsifiers and Their Applications

5.5.2

Emulsion Properties and Protein/Emulsifier Interactions

The formation of the interfacial layer on the emulsion droplet is strongly linked
to the properties of the aqueous environment as well as to the properties of the
oil phase. An example of the former has been provided by Chen and Dickinson
(1995a-c), who showed that the addition of an anionic surfactant, sodium lauryl sulfate (SLES), can introduce flocculation of a gelatin stabilized oil-in-water emulsion at sufficient surfactant concentrations. An increase of the SLES
concentration led to a restahilization of the flocculated emulsion. This could he
related to the hulk behavior, where the anionic surfactant will neutralize the
positively charged gelatin and cause precipitation of the protein. As more surfactant is added, the solubility of the protein increases due to binding of surfactant that increases the protein net charge, and finally the precipitate is
redissolved. Measurements of the composition of the interfacial layer reveal
(Chen and Dickinson, 1995c) that gelatin is initially replaced by SLES, hut as
the surfactant concentration increases, more gelatin can he accommodated at
the interface due to partial neutralization of the gelatin at the interface. This
will eventually cause flocculation to occur (Chen and Dickinson, 1995a). A
further increase of surfactant concentration leads to a decrease in gelatin surface concentration (Chen and Dickinson, 1995c) and a restabilization of the
emulsion (Chen and Dickinson, 1995a). It is noteworthy that an emulsion stabilized by P-lactoglohulin, which was negatively charged under the prevailing
experimental conditions, did not show any signs of flocculation upon addition
of SLES, although a complex was formed in the hulk solution.
Several examples of how the properties of the oil phase composition can affect the structure of the adsorbed layer of protein on the emulsion droplet, and
hence the stability of the emulsion, have been studied. For instance, the work
of Leaver and Dagleish (1992) on the structure of adsorbed layers of ~-casein
on emulsion droplets, where it was found that the cleavage of the protein on
the oil-droplet surface by trypsin gave different products depending on
whether a triglyceride oil or tetradecane, was used. This demonstrates that the
structure of the adsorbed layer depends on the composition of the oil.
Furthermore, it was shown by Heertje et al. (1990) that if a monoglyceride was
dissolved in the oil phase, the displacement of caseinate from the oil/water interface was correlated to the interfacial concentration of monoglyceride. They
also found that at high monoglyceride concentration in the oil phase, the
amount of saturated (monostearoylglycerol) lipid adsorbed at the interface was

Protein/Emulsifer Interactions

135

larger than that of the unsaturated (monooleoylglycerol) one, which also led to
more extensive displacement of caseinate.
We have in the foregoing section mainly been concerned with the binding of
emulsifiers to proteins, but one can also think of cases where there is a repulsion between the lipid and head group and the protein. In this way the structure of the adsorbed protein layer, and hence the properties of the emulsion,
can be changed. This has been demonstrated by Brookbanks et al. (1993), who
found that the thickness of adsorbed layers of milk proteins was significantly
greater when the protein was adsorbed onto negatively charged phosphatidylglycerol (PG) liposomes. The more extended structure was attributed to charge
repulsion between the negatively charged lipid surface and the negatively
charged surface domains of the protein.

5.5.3

Conclusion

The interaction between emulsifiers and proteins is to a larger extent driven by


electrostatic or hydrophobic interactions, or in many cases it is a combination
of the two. Thus, it is commonly observed that ionic emulsifiers interact more
strongly with proteins than nonionic ones. For emulsifiers with low water solubility, e.g., polar lipids, the interaction with proteins is largely dependent on
the phase structure upon addition. The binding can, depending on the type of
emulsifier, lead to stabilization of the protein structure at low-surfactant-toprotein ratios. However, an increase in surlactant concentration can induce
unfolding of the protein and in some cases precipitation of the protein.
We have seen that the stability of emulsions and foams is determined by interfacial processes, which are affected by the properties of the interface as well
as the interactions occurring in bulk solution.
When no emulsifier/protein interactions are present, the composition of the
interfacial film is determined by only the surface activity and concentration of
the components. In the case of reversibility the "race for the interface" is won
by the most surface-active and/or abundant molecule, and in the case of irreversibility the transport rate might also play a role. In this context it has to be
born in mind that proteins can change their conformations (sometimes in a
time-dependent way) at the interface. This may lead to a strong interaction between the protein and the surface, which can hamper the displacement of a
protein by more surface-active emulsifiers.
The presence of protein/emulsifier interactions can have pronounced impact on the interfacial behavior of the components. In cases where the emulsi-

136

Food Emulsifiers and Their Applications

fier binding induces protein unfolding, exposure of hydrophobic domains of


the protein, or precipitation at the interface due to charge neutralization, the
surface activity of the complex is increased compared to the native protein. On
the other hand, if the protein is more soluble or stabilized by the emulsifier interaction, the complex has less tendency to adsorb at the interface.
Precipitation of the complex in the bulk can cause loss of surface-active material and hence a decrease of the surface concentration. The emulsifier/protein
interactions at interfaces can give more efficient packing and thus a higher total surface concentration. If protein/protein interactions take place at the interface, they may be disrupted by protein/emulsifier interactions.
Although emulsions and foams are stabilized by the same mechanisms,
there are marked differences. First, there are profound differences between the
two types of liquid interfaces: the liquid/air and the one between two condensed media. The oil/aqueous interface allows hydrophobic residues to become dissolved in and interact favorably with the oil phase, which is not
possible at the air/water interface. It should be noted that unfolding of the protein induced by action of emulsifiers or by the presence of an interface generally leads to exposure of hydrophobic residues; that is, the unfolded protein is
more "oil soluble" than the native one. Second, in the stabilization of foams
the viscoelastic properties of the surface film as well as the thin aqueous film
has large effects. This means that protein/protein interactions in protein-stabilized foams are important, and the addition of surfactants can disrupt these interactions and lead to the collapse of the foam. Furthermore, they can also be
stabilized by low molecular weight emulsifiers by means of Gibbs and
Marangoni effects.
Steric and/or repulsive forces are important for stabilization of emulsions.
Therefore, the mixed-protein/emulsifier layer should be optimized with respect
to charge and/or by segments in the surface layer protruding into the aqueous
environment to give a hairy structure that will sterically stabilize the emulsion.
This chapter has shown the enormous variety in emulsifier/protein interactions that can occur in food emulsions and foams. Each protein/emulsifier
combination is unique and its behavior specific when applied in a particular
foam or emulsion, where other ingredients are present. However, we have
demonstrated that it is possible to establish certain principles for
protein/emulsifier interactions. These principles based on mechanisms at the
molecular level have also to be transferred to processes of manufacturing, storage, and distribution of food products based on emulsions and foams. The

Protein/Emu1sifer Interactions

137

challenge is to increase the resistance of microbial growth without excessive


use of antimicrobial substances, control digestion of the product, and achieve
controlled release of flavors as well as design new functional ingredients based
on natural products.

Acknowledgement
Thomas Arnebrant thanks the Swedish Research Council for Engineering
Sciences (TFR) and Martin Bos acknowledges the EU Human Capital and
Mobility program (ERBCHRXT930322) for their financial support. Tommy
Nylander acknowledges the Swedish Council for Forestry and Agricultural
Research. David Clark acknowledges the support of the Biotechnology and
Biological Sciences Research Council.

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3880-3885.
Steinhardt, J., Stocker, N., Carroll, D., et al. (1974). The non specific binding of amphiphiles by proteins, Biochemistry, 13, 4461-4468.
Su, Y.-Y.T., Jirgensons, B. (1977). Further studies of detergent-induced conformational
transitions in proteins, Arch. Biochem. Biophys., 181, 137-146.
Subramanian, M., et al. (1986). Interaction of proteins with detergents: Binding of
cationic detergents with lysozyme,]. Biosci., 10, 359-371.
Tabata, Y., lkada, Y. (1990). Adv. Polym. Sci., 94, 107.
Tadros, Th. F., Vincent, B. (1983). In Encyclopedia of Emulsion Technology (ed. P.
Becher), Marcel Dekker, New York, Vol. 1, p. 129.
Tanford, C. (1967). Physical Chemistry of Macromolecules, Wiley, New York.
- - - (1980). The Hydrophobic Effect: Formation of Micelles and Biological
Membranes, Wiley, New York.
Tiberg, F. (1994). Self-Assembly of Nonionic Amphiphiles at Solid Surfaces, PhD
Thesis, University of Lund, Sweden.
Tilton, R.D., et al. (1993). Effect of anionic surfactant on interactions between lysozyme
layers adsorbed on mica, Langmuir, 9, 2102-2108.
Townend, R., et al. (1969). The state of amino acid residues in ~-lactoglobulin, Arch.
Biochem. Biophys., 129, .567-580.
Verger, R., Pattus, F. (1982). Lipid-protein interactions in monolayers, Chem. Phys.
Lip., 30, 189-227.
Wahlgren, M. (1992). Adsorption of Proteins and Interaction with Surfactants at
Solid/Liquid Interfaces, PhD Thesis, University of Lund, Sweden.
- - - , Arnebrant, T. (1990). Adsorption of ~-lactoglobulin onto silica, methylated silica and polysulphone, ]. Colloid Interface Sci., 136, 259-265.
- - - (1991). Interaction of cetyltrimethylammonium bromide and sodium dodecylsulphate with ~-lactoglobulin and lysozyme at solid surfaces, ]. Colloid Interface
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- - - (1992). The concentration dependence of adsorption from a mixture of ~-lac
toglobulin and sodium dodecyl sulphate onto methylated surfaces, ]. Colloid
Interface Sci., 148, 201-206.
- - - et al. (1993a). Adsorption of globular model proteins to silica and methylated
silica surfaces and their elutability by dodecyltrimethylammonium bromide,
Colloids Surf, A, 70, 139-149.
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akyltrimethylammonium bromides, Colloids Surf A, 70, 1.51-1.58.
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23-31.
Wallin, R., Arnebrant, T. (1994). The activity of lipase at the cubic liquid-crystalline
phase/water interface,]. Colloid Interface Sci., 164, 16-20.

146

Food Emulsifiers and Their Applications

Wallin, R., et al. (1993). Stabilisation of glucose oxidase by entrapment in a cubic liquid-crystalline phase, Biocatalysis, 8, 73--80.
Walstra, P. (1983). In Encyclopedia of Emulsion Technology (ed. P. Becher), Marcel
Dekker, New York, Vol. 1, p. 57.
Wannerberger, K., et al. (1995). Adsorption onto methylated silica surfaces from lipase/surfactant solutions, Colloids and Surfaces B: Biointerfaces. In press.
Watts, A. (ed.) (1993). Protein-Lipid Interactions, New Comprehensive Biochemistry,
Vol. 25, Elsevier, Amsterdam.
Welin-Klintstrom, S., et al. (1993). Surfactant and protein interactions on wettability
gradient surfaces,]. Colloid Interface Sci., 158, 188-194.
Wilde, P.J., Clark, D.C. (1993). The competitive displacement of ~-lactoglobulin by
Tween 20 at the oil-water interface,]. Colloid Interface Sci., 155, 48-54.
- - - , Marion, D. (1993). The influence of competitive adsorption of a lysopalmitoyl
phosphatidylcholine on the functional properties of puroindoline, a lipid binding
protein isolated from wheat flour,]. Agric. Food Chem., 41, 1570-1576.
De Wit, J. N. (1989). Functional properties of whey proteins, in Developments in Dairy
Chemistry-4 (ed. P.F. Fox), Elsevier Applied Science, London, pp. 285-322.
Wangnerud, P. (1994). Adsorption oflonic Surfactants on Charged Solid Surfaces, PhD
Thesis, University of Lund, Sweden.

SIX

Physicochemical Aspects of
an Emulsifier Functionality
Bjorn Bergenstahl

6.1

Introduction

The characteristic property of all emulsifiers is their surface activity. Surface


activity is the ability to form a surface excess at interfaces. The formation of
adsorbed layers at interfaces are displayed in a change of a range of easily observable and technically important properties.
l. The surface tension is reduced.

2. The lifetimes of bubbles are increased. (Only very pure water displays a
very short lifetime, a few seconds, of bubbles created by shaking.
Normal standard "pure water," double distilled, usually displays a bubble lifetime of about 20 to 30 seconds.)
3. The emulsifiability of oils in water is enhanced. Smaller drops with a
longer lifetime are formed with less stirring.
4. The aggregation rate of dispersed particles is changed. Surface-active
additives may induce or prevent flocculation of disperions.
5. The sediment volume of settling particles is influenced. Surface additives inducing adhesive may create a loose or compact sediment.
6. Crystallization properties are changed. This may include crystallization
rate and crystal shape.

147

148

Food Emulsifiers and Their Applications

This chapter aims to discuss the principal physical origin of the various functionalities of typical lipid food emulsifiers. Aspects on the functionality under
very different conditions in various foods will be discussed. I will try to show how
we may select emulsifiers on the basis of their fundamental properties.

6.2

Surface Activity

When an additive is added to a solution, the gain of entropy is very large at low
concentrations. If the additive displays surface activity and adsorbs at an interface, the system loses entropy, which has to be balanced by a gain in free
energy due to the adsorption. At very low concentrations the solubility always
prevails, but when the concentration is increased, more and more of the available surfaces will be covered by the adsorbed molecules. To display surface
activity, the emulsifier needs to have certain properties:
1. It has to form a noncrystalline form t in contact with water.
2. It should have a reduced solubility in water due to a large hydrophobic part.
3. It has to interact with water through polar interactions.
4. It should have a significant molecular weight to reduce the effect of the reduced entropy when it adsorbs.
5. It has to have a reduced solubility in an oil environment due to large size and
the presence of polar groups at the interface.
High-melting emulsifiers do not display surface activity when dispersed in water until a critical temperature, the Krafft temperature, has been reached. At this
temperature the emulsifier solubility in the solution has reached a sufficient concentration to allow for a significant formation of adsorbed layers at the interfaces.
The presence of hydrophobic parts of the molecules increases the energy
gain due to adsorption. In aqueous environments most emulsifiers tend to aggregate in micelles at a critical concentration, erne (critical micelle concentration), or to precipitate as liquid crystals. Above the aggregation concentration
all properties depending on the chemical potential, for instance the adsorption
properties, are more or less constant. The aggregation is mainly driven by the
presence of the hydrophobic parts of the molecules (Tanford, 1973).
t Several lipid emulsifiers are exceptions and are applied in a hydrated gel form (a crystals).
However, this crystal form resembles the liquid crystalline form in terms of interactions with
both phases and spreadability over the interfaces.

Physicochemical Aspects of an Emulsifier Functionality

149

A polar part of the molecule is necessary to avoid the formation of a separate oil phase. The type of aggregates formed during the adsorption will reflect
the balance between the polar part and the hydrophobic part of the molecule.
The free-energy gain at adsorption is mainly proportional to the molecular
weight, while the entropy loss due to the demixing is independent of molecular
weight. Hence, small molecules, for instance lower alcohols, do not form adsorbed layers at hydrophobic surfaces in contact with water solutions, while
pronounced layers are formed with additives of higher molecular weights, for
instance monoglycerides. Proteins display a much higher surface activity than
protein hydrolysates.
In an oil environment, solvophobic effects are absent and the adsorption
has to be generated by polar interactions between the second phase and the
surface-active molecule.
The interaction between particles is influenced when the particles are covered by an adsorbed layer of an emulsifier. The change in the interaction
strongly influences the macroscopic properties of the dispersions (Table 6.1).
Table 6.1

Effects of changes in the interactions on the


macroscopic properties of dispersions

Interaction

Stability

Sedimentation

Attraction
Repulsion

Flocculation
Stable

Large sediment volume


Small sediment volume

The solution properties of emulsifiers are determined for the surface activity of the emulsifiers. In addition, the ability to generate repulsive interactions
is also reflected in the solution properties of emulsifiers.

6.3.

Solution Properties of Emulsifiers

When water is added to a surfactant system, the solubilization in the system


may in principle pass through a series of aggregation structures and phases in
a particular sequence. The sequence is: reversed micelles ~ reversed hexagonal phase ~ lamellar phase ~ hexagonal phase ~ micellar solution ~ molecular solution (Fontell, 1978) (Figure 6.1).
The free energy of solubilization, LlG,oluhilization' can be described as a sum of

150

Food Emulsifiers and Their Applications

reversed
micelles
reversed
hexagonal
lamellar phase
phase

Melting point

M---

hexagonal
phase
,
:micelles

'
'
Krafft
point

Emulsifier

Figure 6.1 A typical sequence of liquid-crystalline phases and solution phases


formed in a binary emulsifier mixture. (Modified after Fontell, 1978.)

free energy contributions in the process by the expression: emulsifier phase +


water

more solubilized phase:

L\Gphase transformation

.L\.Gmixing

+ .AGpolar group/water interaction + .L\.~ydrophobic

where .ilGmixing is negative when changing from large aggregates to small aggregates (micelles and molecular solutions).
is positive and equal to Ahydrocarbonlwater 'Y hydrocarbon/water The hydrophobic effect is the driving force for the aggregation and gives the upper

ilGhydrophobic

limit of the molecular solubility for amphiphilic molecules (critical micelle


concentration) .
is negative. This term consists mainly of the work released
when more water allows a larger separation between repelling aggregates or
.ilGpolar group/water

molecules:

L l I

next aggregate

LlGpolar group/water

F(l) dl

all neighbors

aggregate

where l is the average distance between the polar groups and F(l) is the interaction.
The area per molecule in the aggregates is given by the balance between
the interfacial tension of the oil/water interface and of the space needed for the

Physicochemical Aspects of an Emulsifier Functionality

151

polar group itself and the space generated by repulsive interactions between
the emulsifier head groups at the interface.
The area per molecule expands in the series Areversed micelles < Areversed hexagonal

< Alamellar Ahexagonal < Amicelles"


At a specific ratio of water and emulsifier, the system's tendency is to obtain
aggregates as small as possible to maximize the AGmixing and the AGpolar group/water
The lower limit in aggregate size is given by the increased hydrophobic contact
between the exposed hydrocarbon/water interface.
The interesting result of this exercise is that the area per molecule is to a
large extent a measure of the ability to generate repulsive interactions.
In the solubilization sequence, reversed aggregates
hexagonal

phase~

micellar

solution~

lamellar phase

molecular solution, the area per mol-

ecule of the surfactant/water interface increases. Depending on the packing


constraints given by the hydrophobic moiety in the aggregates, the range of the
repulsive interaction on the polar side of the molecule, and the molecular
weight, this process has to proceed more rapidly or more slowly (Israelachvili
et al., 1976, 1977). Hence, the packing constraints of the hydrocarbon chain
are an important link between properties and aggregation.
The ratio of the actual area A, as it is created by the repulsive interactions,
to the theoretical area of a saturated hydrocarbon chain, A0 (23

A2)

enforces

different geometries (Israelachvili et al., 1976, 1977) due to the different ratio
of volume to area of different aggregates, as shown in Table 6.2.
The successive solvation of surfactants in Table 6.2 correspond to a successive change into aggregates that correspond to a more long-range interaction. If there is an upper limit for the repulsion, the solvation series is
terminated at that stage. Hence, the maximum solvated aggregate formed at a
surplus of water is a measure of the ability of the emulsifier to generate repulsive interactions.
The area of the molecule is a measure of the interaction when water is
available, and may be generalized as the hydrophilicity of the molecule. The
spatial requirement of the hydrophobic part of the molecule is of course a measure of the hydrophobicity of the molecule. Consequently, there is a close link
with the classical view of emulsifiers as molecules with a balance between the
hydrophobic and the hydrophilic properties, as they are expressed in the HLB
numbers, proposed by Griffin (1949, 1979).

152

Food Emulsifiers and Their Applications

Table 6.2

The geometries of different aggregates


Packing constraint t
(A 0 = 23A 2 for a
saturated hydrocarbon tail)

Area/volume
Sphere (micellar solution)
2

2 1tr

(4/3)1tr

= 3Ao

3 Vhydrophoh
rhydrophob

Rods (hexagonal phase)

2r1tl

n. /z

2 vhydrophob

rhydrophob

2A

Bilayers (lamellar phase)

Vhydrophoh _

rhydrophob

Reversed rods (reversed hexagonal phase)

radius of the aggregate, usually limited by the length of molecule

= a fictitious length of the aggregate


vhydrophob

= volume of the hydrophobic part of the molecule

rhydrophob

the maximum length of the hydrophobic moiety

Ao
A

area of a cylindrical packing of the hydrophobic moiety

(= V hydrophob 1 rhydrophob or 23 A2 per hydrocarbon chain)


area per molecule at an average water/amphiphilic interface

t The packing constraint is here defined as the necessary cross section of an amphiphilic molecule
in the aggregate at the oil/water interface. This definition is Aofpacking parameter according to
Israelachvili et al. (1992, 1976, 1977).

153

Physicochemical Aspects of an Emulsifier Functionality

6.4 The Use of Phase Diagrams


to Understand Emulsifier Properties
Friberg and coworkers (Wilton and Friberg, 1971; Friberg and Mandel, 1970b;
Friberg and Rydhag, 1971; Friberg and Wilton, 1970; Rydhag, 1979; Rydhag
and Wilton, 1981; Friberg et al., 1969; Friberg and Mandel, 1970a; Friberg,
1971) have investigated phase diagrams and emulsion stability extensively.
They concluded that the optimum composition for a stable emulsion should be
that at which the lamellar phase, the oil phase, and the water phase are in
equilibrium in the corresponding phase diagram (Figure 6.2).
The relation between the formation of lamellar phases and emulsion stability is basically of an empirical nature. The emulsifiability is enhanced at certain compositions (Friberg and Mandel, 1970b; Friberg and Rydhag, 1971;
Friberg and Wilton, 1970), and the formation of crystalline phases corresponds
to an observed destabilization (Wilton and Friberg, 1971). The formation of
multilayers around the emulsion droplets under certain conditions has also
been shown (Friberg, 1990).

It was suggested that the formation of a multilayer of a lamellar liquid-crys-

p- Xylene
Micellar solution and
oil phase
lamellar micellar solution
and oil phase

Water

Nonylphenoi-E09

Figure 6.2 Emulsion experiments in the phase diagram of an ethoxylated nonyl-phenol


and xylene. Systems with compositions corresponding to the position in the phase diagram were weighed into flame-sealed ampoules. The emulsifiability of the systems was
tested by shaking the ampoules. The stability of the emulsions formed was observed the
emulsification. (Modified from Friberg et al., 1969; Friberg and Mandell, 1970a.)

154

Food Emulsifiers and Their Applications

talline phase coating the droplet surface reduces the van der Waal's attraction
and that this was an important contribution to the observed effects in the emulsification experiments (Friberg, 1971). However, this explanation is not a useful general explanation since the emulsifier concentration in optimized food
emulsions rarely is high enough to allow for multilayer adsorption (Walstra,
1988; Dickinson, 1986). Obviously, this observation is contradictive to the
need for a separate phase of liquid-crystalline material around the droplet.
However, a correlation between the presence of, or the possibility to form, liquid-crystalline phases and emulsion stability is still experimentally observed
in several systems. To stabilize an dispersion, the emulsifier should:

l. Contribute to the repulsive interactions between the droplets


2. Contribute to the interfacial viscosity
3. Be well anchored to the interface
These properties are reflected in the formation of various liquid-crystalline
phases (Table 6.3). These aspects are illustrated by a few examples.
Table 6.3

The relation between the function of an emulsifier to stabilize an


emulsion and its ability to form various aggregation structures

Stabilizing property
aggregates

Micelles

Bilayers

Reversed

Water-continuous emulsions
Repulsive interactions
Interfacial viscosity
Anchoring

Optimal
Weak Optimal
Too water-soluble

Intermediate
Weak
Optimal

Weak
Acceptable

Oil-continuous emulsions
Repulsive interactions
Interfacial viscosity
Anchoring

Weak
Weak
Acceptable

Intermediate
Optimal
Optimal

Optimal
Weak
Too oil-soluble

6.5 Examples of the Relation between


Phase Diagrams and Emulsion Stability
6.5.1 Monoglycerides
A technical monoglyceride at room temperature remams m a nonhydrated
crystalline phase (~ phase) in equilibrium with a surplus of water. Above

Physicochemical Aspects of an Emulsifier Functionality

155

40C, the monoglyceride takes up water and a lamellar phase is formed


(Wilton and Friberg, 1971). The lamellar phase coexists with a surplus of water (no micelles are formed). When the lamellar phase is cooled, a semicrystalline phase, termed "a phase," is formed. This phase is metastable below
30C and converts only slowly into an aqueous and a ~ phase.
The swelling of the lamellar and a phases indicates the existence of a strong
repulsive hydration force. This force has been measured by the osmotic stress
technique (Figure 6.3). In contrast, no hydration force strong enough to separate
the bilayers is present in the ~ phase. The hydration force between emulsion
droplets coated with this emulsifier depends on the liquid-crystalline state of the
adsorbed emulsifier film in the same way. This explains why monoglycerides appearing in the ~form are inactive as emulsifiers, and why a monoglyceride-stabilized emulsion rapidly destabilizes when the monoglyceride converts from
lamellar or a into~ phase (Wilton and Friberg, 1971). In technical systems, it is
important that the conversion of a phase into ~ phase is delayed. An a phase
can be stabilized by the presence of ionic charges (soap) (Larsson and Krog,
1973) or by a wide distribution of the fatty acid-chain composition. The solution
properties of a range of food emulsifiers are summarized in Table 6.4.

6.5.2

Lecithins

Lecithin is one of the most commonly used food emulsifiers, and its popularity
can be expected to grow even further due to its natural origin. Technical

lecithins, usually soybean lecithin, are always natural mixtures of various


phospholipids. The most frequent one is phosphatidylcholine (PC). The second

'"_

0..

s
.5

";:;'"
"'"0..
:n">

..

c
ccccc

c
c c

"

0..

Lamellar phase 60 C

c Gel phase 23 C

c
00

:;

-1
0

10

15

dw(A)

Figure 6.3 The hydration repulsion between bilayers of monopalmitin in the liquidcrystalline and gel states. (Redrawn from Pezron et al., 1991.)

156

Food Emulsifiers and Their Applications

Table 6.4

Formation of liquid-crystalline phases by lipid emulsifiers


Fatty
acid

Liquid-crystalline
phases formed at

Upper swelling
limit (at 25C)

Distilled saturated

C18-16

50%

Krog, 1990

Distilled unsaturated

Cl8:l-2

35%

Krog, 1990

Monoolein

C18:1

Lamellar phase at 50C


Cubic at 70C
Cubic< 20C
Reversed hexagonal at 55C
Cubic< 20C
Reversed hexagonal at 90C

40%

Krog, 1990

Emulsifier
Monoglycerides:

Tetraglycerolesters:
Tetraglycerol
monolaurin

C12

Lamellar < 20C


Fluid isotropic 40C

55%

Krog, 1990t

C16-l8

Lamellar 45C

55%

Krog, 1990

Reversed hexagonal at 45C


Lamellar at 42C

40%
60%

Krog, 1990
Krog, 1990

Organic acid esters:


Diacetyl tartaric acid
monoglyceride ester

Sodium steraoyllactylate:
pHS
pH7

C18
C18

Sorbitan esters:
Polyoxyethylene (20) C18:1
sorbitan monooleate
Polyoxyethylene (20) C18
sorbitan monostearate
Sorbitan stearate

C18

Hexagonal phase (up to 30C)


aud micellar solution
Hexagonal phase (30 to 50C)
and micellar solution
above 30C
Lamellar above 50C
-

Hall, Pethica, 1967


Hall, Pethica, 1967
Hall, Pethica, 1967

tThe data are extracted from a review of several original sources.

is phosphatidylethanolamine (PE). Phosphatidylinositol (PI) and phosphatidic


acid (PA) are usually present at intermediate levels, and phosphatidyl serine
(PS), lysophosphatides (LPC and LPE), etc., at low levels. Nonphosphatides
such as steroids, vitamin E, and free fatty acids are usually also present in
technical products. The properties of lecithins reflect some type of average
properties of the mixture. This section will first describe the characteristic

Physicochemical Aspects of an Emulsifier Functionality

157

properties of the most common phosphatides and then discuss the properties of
various mixtures.

6.5.3

Phosphatidylcholine

The phase diagram of a typical unsaturated phosphatidylcholine is displayed


in Figure 6.4. The phase diagram is characterized by a large swelling lamellar
phase. Saturated phosphatidylcholines have a phase transition temperature up
to about 40C, whereas the corresponding temperature for unsaturated
lecithins is well below 0C. The phase diagram of soybean PC is described in
Bergenstahl and Fontell (1983) and is rather similar to the phase diagram of
dioleoyl PC.

6.5.4

Phosphatidylethanolamine

Phosphatidylethanolamine is less hydrophilic than PC. The saturated


ethanolamines form lamellar phases that swell less than the corresponding PC
species. The phase transition temperature is about lO to 40C above the corresponding temperature of the phosphatidylcholine (Figure 6.5). The more limited ability to create long-range repulsive interactions, and thereby to defend a
large molecular area, is displayed in the tendency to form reversed hexagonal
phase with unsaturated PE species, as shown in Table 6.5.

200

Reversed
Hexagonal

~ 150

8 100
~

50

Cubic
Crystalline
phase

Water

Figure 6.4.

Lamellar
phase

Dioleoylphosphatidylcholine

The phase diagram of water and dioleoylphosphatidylcholine. (From

Bergenstahl and Stenius, 1987.)

158

Food Emulsifiers and Their Applications

90

G
~

c.

E
~

[J

70

[J

50

[J

30

10
-10
10

12

14

16

18

20

22

Chain length

Figure 6.5 The main transition temperature for phosphatidylcholine (PC) () and
phosphatidylethanolamine (PE) (D) as a function of chain length. The sources of data
are given in Table 6.4.

6.5.5

Phosphatidylinositol

The phase diagram of soybean PI and water has been determined by the author
(1991) and by Soderberg (1990). The diagram is characterized by a large
lamellar phase with an unlimited swelling. The liquid-crystalline phase is
formed below room temperature.

6.5.6

Phosphatidic Acid

The phase diagram of the sodium salt of dioleoylphosphatidic acid has been determined by Lindblom et al. (1991). The phase diagram is characterized by a
lamellar phase that transforms to a reversed hexagonal phase at about 30% of
water. This transformation occurs although there is an ionic charge on the molecules and despite the small head group. A possible explanation, supported by
evidence from NMR measurements, is that this is due to ion condensation.

6.5.7

Lysophosphatides

The phase diagrams of a series of different lysophosphatides has been investigated by Arvidsson et al. (1985). Lysophosphatidylcholine has the same hydrophilic polar group as the ordinary PC but only one of the two fatty acids.
This reduces the volume demand of the aggregate, and the packing constraint
allows for the formation of micelles and hexagonal phases.

6.5.8

The Properties of Mixtures of Phosphatides

Technical phosphatides are always mixtures. Their properties reflect some


type of average that the mixture develops. One way to investigate this is to de-

159

Physicochemical Aspects of an Emulsifier Functionality

Table 6.5

The formation of liquid-crystalline phases by various phospholipids

Phospholipid

Fatty
acids

Liquid-crystalline
phases formed at

Upper swelling
limit (at 25C)

Lamellar phase at 55C


Lamellar phase at 4I C
Lamellar phase at 23 C
Lamellar below 0C
Lamellar at 2C
Lamellar below 0C

36%
40%
42%
44%
35%

Phosphatidylcholine:
Distearoyl
Dipalmitoyl
Dimyristoyl
Dioleoyl
Egg PC
Soybean PC

CIS
CI6
CI4
CIS:!
CI6-IS:I
CIS:I-2

Small, I9S6t
Insko & Matsui,I97S
Janiak et al., I97S
Bergenstahl & Fortell, I9S7
Small, I9S6
Bergenstahl & Fortell, I9S7

Phosphatidyletanoleamine:
Dipalmitoyl

CI6

Dioleoyl

C1S:l

Soybean PE

CIS

20%
Lamellar phase at 68C
Reversed hexagonal at S4C
Lamellar below 0C
20%
Reversed hexagonal at 5C
l-2+ Reversed hexagonal
30%
above 0C

Caffrey, I9S5
Gawrish et al, 1992
Bergenstahl, 1991

Phosphatidylinositol:
Soybean PI

C1S:I-2+

Lamellar below 0C

Unlimited

Bergenstahl, 1991
Soderberg, 1990

ClS:l

Lamellar below 0C

Unlimited

Lindblom et al., I99I

CI6

Micellar solution below 0C

Unlimited

Eriksson et al., I9S7

Phosphatidic acid:
Dioleoyl
Lyso PC:
Palmi toy!

iThe data are extracted from a review of several original sources.


+Mainly.

terrnine the type of liquid-crystalline phase that develops when different phosphatides are allowed to interact together with water. Figure 6.6 shows the
phase diagram of dioleoyl PC and dioleoyl PE in 40% water (Eriksson et al.,
1985). The figure shows that a lamellar phase is formed when the system contains mainly PC, but that about 60% PE nonlamellar phases start to form. This
change is enhanced at high temperatures. Between the hexagonal phase and
the lamellar phase is an area in which a cubic phase appears (above 50C).

160

Food Emulsifiers and Their Applications

100
Cubic phase

80

Reversed
hexagonal
phase

e
::l

60

'"<l.l

Lamellar phase

0.,

40

20

100%
DOPE

50150

100%
DOPC

Composition
Figure 6.6

The phase diagram of dioleoyl PC and dioleoyl PE with 40% water.

(Redrawn after Eriksson et al., 1985.)

The more highly unsaturated soybean PE and soybean PC display a similar


aggregation pattern, but the temperature at which the system changes from
lamellar to nonlamellar phases is lower (Figure 6. 7), and the phase diagram is
dominated by the hydrophobic properties of the PE up to fairly high concentrations of PC. A mixture of PI and PC displays the extreme swelling properties of
ionically charged emulsifiers at an early stage. This was indeed also expected
since a similar pattern was observed when a small amount of ionically charged
detergents was added to the lamellar phase formed by monoglycerides
(Larsson and Krog, 1973). When PI and PE are mixed, the properties of the
mixture are dominated by the hydrophilic PI up to quite a high PE:PI ratio.
A preliminary conclusion from this work is that the properties of phosphatide mixtures are determined by the ratio of anionic (particularly PI) phosphatides to PE rather than by the PC:PE ratio.
Technical soybean lecithin contains a mixture of different phospholipids
1979). In most cases, the weakly hydrophilic phosphatidylethanolamine dominates, and this type of lecithin is suitable for inverse
(Rydhag,

emulsions such as in margarine. More hydrophilic soybean lecithins suitable

Physicochemical Aspects of an Emulsifier Functionality

Water

PC

161

Water

PI

Water

Figure 6.7 The phase diagram of soybean PC, soybean PE, and water; of soybean PC,
soybean PI, and water; and of soybean PI, soybean PE, and water. (Redrawn after
Bergenstahl, 1991.) The cubic phase was not included in the original drawing, but it is
a possible interpretation of the x-ray peaks included in the paper. It is also supported
by the data from the study by Eriksson eta!. (1985).
for oil-in-water emulsions are obtained by partial hydrolysis to form
lysolecithins (Emulfluid E). t It is also possible to increase the effective hydrophilicity of the PE by making the polar head group larger through acetylation (Emulfluid A).

6.6

Some Ways to Classify Emulsifiers

A common problem in industrial development work is the choice of suitable


surfactants to obtain the desired results. In the literature a number of different
methods of making a fast preliminary selection of suitable emulsifiers have
been proposed. The most common methods and concepts are discussed here
and are compared with the function of the emulsifier in the emulsion.

6.6.1

The Solubility Concept

One of the first ideas, proposed by Bancroft (1913) at the beginning of the century, was that the solubility of the emulsifier determines the type of emulsion
that is formed. An oil-soluble emulsifier will create an oil-continuous ernult Emulfluid, Lucas Meyer, Elbdeich 62, Hamburg, Germany

162

Food Emulsifiers and Their Applications

sion, and a water-soluble emulsifier will tum the emulsion into a water-continuous one. This is true for low molecular emulsifiers with a high solubility (usually in micellar aggregates), but it is also valid for polymers. However, most
likely, the concept can also, to some extent, be expanded to include emulsifiers
with just a dispersibility in either one of the phases (for instance lecithin).
Experience in this direction is exemplified in Table 6.6. However, the Bancroft
rule provide us just with the first very general directions. To proceed further we
need possibilities to rank emulsifiers quantitatively.
Table 6.6 Emulsifiahility compared with solubility according to the Bancroft
rule. (Ostberg et al., 1995.)
Emulsifier

Soluhility/dispersihility

Type of emulsion

Sorbitan esters
(Span)
Etoxylated sorbitan
esters (Tween)
Hydrophobic lecithin
(normal technical lecithin)
Hydrophiliclecithin
(high LPC or low PE)
Proteins
Fat crystals

Oil-soluble

Oil-continuous

Water-soluble

Water-continuous

Oil-dispersible

Oil-continuous

Water-dispersible

Water-continuous

Water-soluble
Oil-dispersible

Water-continuous
Oil-continuous

6.6.2 The Phase Inversion Concept


Ethoxylated surfactants have a tendency toward declining hydrophilicity with increasing temperature. This leads to a change from water solubility at low temperatures to oil solubility at higher temperatures. According to the Bancroft rule,
this will cause a given system to switch from being water-continuous to being oilcontinuous. The hydrophilicity can be viewed as a property that is gradually lost
with increasing temperature. The distance from the breakeven point, the phaseinversion temperature (PIT), is then a measure of the strength of the hydrophilicity. Shinoda claims that the best stability of an oil-in-water emulsion is obtained
at 30C below the PIT and for a water-in-oil emulsion at about 20C above the
PIT. However, the droplet size obtained directly after the homogenization (by
shaking) reach a minimum at the PIT. Consequently, Shinoda suggests that the
emulsifier should be chosen so that the emulsification can be performed at a PIT

Physicochemical Aspects of an Emulsifier Functionality

163

about 20 to 30C above the final storage temperature [emulsification by the PIT
method (Shinoda and Saito, 1968)].
Shinoda and coworkers (Shinoda and Saito, 1968; Shinoda and Kunieda,
1983; Kunieda and Ishikawa, 1985, reviewed in Shinoda and Friberg, 1986)
have worked according to this concept and characterized a number of different
ethoxylated emulsifiers in combination with various solvents. They then found
that the PIT depends not only on the number of ethoxy groups but also on the oil
phase, indicating the importance of the solubility properties for the stability.
Emulsification experiments performed with a range of different oil-to-water
ratios show that the emulsion type is determined mainly by the emulsifier
properties and is for many systems (pure solvents!) very insensitive to the
phase ratio (Shinoda and Friberg, 1986).
It is obvious that this says a lot about the properties of ethoxylated surfactants
but its applicability to food emulsions is very limited for two main reasons:
1. The concept is based on strongly temperature-dependent properties of
the emulsifiers. This excludes ionic emulsifiers (less important for the
food industry), and it also excludes the most commonly used polyhydroxy and nonionic zwitterionic emulsifiers as they display a very weak
temperature dependence of their hydrophilicity.
2. The solvent properties are important in the PIT concept. However, food
emulsions are made almost solely from triglyceride oils and water that will
behave differently due to the large molecular weight of the oil molecules.

6.6.3

The HLB (Hydrophilic/Lipophilic Balance) Concept

Emulsifiers are molecules with a duality in their properties. The balance between the hydrophobic and hydrophilic properties of the molecules should
then determine the performance, for instance to the type of emulsion formed. If
the emulsifier is changed from being hydrophobic to hydrophilic, the emulsion
formed changes from oil-continuous to water-continuous. The balance of the
emulsifier is recorded as a number, the HLB value. When this concept was introduced by Griffin (1949), the HLB value of unknown emulsifiers was determined by comparing the emulsification properties in a predetermined system
of a mixture of hydrophobic and hydrophilic emulsifiers with a predefined
HLB number.
The important development of the HLB system came when the group contribution system was constructed by Davies (195 7), and it became possible to

164

Food Emulsifiers and Their Applications

estimate an HLB value of an unknown emulsifier from the molecular formula


(Table 6.7). The advantage of the HLB concept is that it makes it possible to
characterize numerous emulsifiers and emulsifier blends (it is usually assumed
that it is possible to calculate an average HLB value from the w/w composition). Large tables of data for commercial emulsifiers are also available.
The limitation of the HLB value is that it provides a rather one-dimensional
description of the properties (molecular weight and temperature dependence
are omitted). It is also difficult to calculate useful HLB values for several important food emulsifiers, for instance phospholipids. The HLB values do not
include the important crystallization properties of monoglycerides and modified monoglycerides.
Table 6.7 Calculation of HLB numbers according to Davies (1957). The table is modified according to Davies (1957). HLB = 7+L
group contributions. (From Bergenstahl and
Claesson, 1990.)
Group

Group contribution

Carboxylic acid soap


Sorbitan ester
Glyceryl ester
Ester
Carboxylic acid
Alcohol
Ether
EO group
CH3 , CH2 , CH

212
6.8
5.25
2.4
2.1
1.9
1.3
0.33
-0.475

6.6.4 A Comparison between the HLB and the Geometry


of the Molecule
There is an obvious analogy between the idea of a hydrophilic lipophilic balance and that of a balance in the molecules that are appearing in the packing
constraints creating the different association structures (Figure 6.8). Griffin
(1978) has also suggested a relationship between various solution properties
and the HLB number. Transforming these descriptions into various aggregation
structures, a clear relation between the molecular packing and the HLB value
is obtained.

Physicochemical Aspects of an Emulsifier Functionality

Aggregate

Shape

~>t
..,.,--.

165

Character in solution

HLB

Clear solution

> 13

>2

White dispersion
Milky appearance

7-10

l-2

Lumps of emulsifier

<7

<1

Micelles

Lamellar phase

Reversed aggregates

Figure 6.8 A comparison between molecular aggregation, solution characteristics,


AIA 0 , and the packing parameter. (Modified from Bergenstiihl and Claesson, 1990.)

This result shows that the ability to form liquid-crystalline phases corresponds to the traditional HLB characterization of the emulsifiers.

6.6.5 The Role of the Emulsifier in Homogenization


The discussion so far has been dealing mainly with the situation when droplets
are protected by a layer of emulsifier. However, the emulsifiers also have a crucial role during the emulsification that usually is included in all empirical
tests that are the bases for the rules.
When an emulsion is created from a large and homogeneous oil phase, the
emulsifier should support two different processes: the formation of new droplets
and protection against recoalesce. The emulsifier acts according to both static
and dynamic (diffusion-induced) interactions (Walstra, 1983) (Table 6.8).
Table 6.8

The role of the emulsifiers during the formation


of emulsions
Static

Dynamic

interfaces

Interfacial
tension

across the interfaces

Stabilize the
droplets

Repulsive
surface forces

Diffusion to the
the interfaces

Destabilize the

Diffusion to and

166

Food Emulsifiers and Their Applications

The principal role of the intenacial tension is obvious. The presence of emulsifiers lowers the intenacial tension from about 30 mN/m for a triglyceride/water
system to between 1 and lO mN/m. Nonionic emulsifiers close to the PIT create
densely packed intenaces with very low intenacial tensions. However, the effects of the intenacial tension itself are not very large. Walstra (1983) has shown
that the droplet size is only weakly dependent on the intenacial tension.
During the homogenization, new intenaces are formed. The emulsifiers have
to diffuse to the intenaces to lower the intenacial tension during the events when
the droplets are formed. This process must be rapid to be successful, as rapid as
the time scale for the formation of the droplets. For geometrical reasons, the diffusion from the surrounding phase of the droplet is much more rapid than the
diffusion from the internal liquid. This is one important contribution to the validity of the solution rules (Bancroft, PIT, HLB, and phase diagrams).
During the homogenization, the water-soluble substances in the oil phase
diffuse over to the water phase. These types of diffusion across the intenaces
create disturbances that contribute to the emulsification. In many systems, this
effect gives an increased efficiency if the emulsifier is added to the oil phase
before the emulsification. For dispersible emulsifiers (phospholipids) there are
also other reasons why it is more efficient to add the emulsifier to the oil phase
instead of the water phase. During the homogenization, phospholipids tend to
form stable liposomal dispersion in competition to the emulsification of the oil
phase. Westesen has indeed observed that a significant fraction of the phospholipids in a commercial phospholipid emulsion for paranteral use is lost in
liposomal aggregates (Westesen and Wehler, 1992).
Emulsification involves an intensive shear. The shear by itself causes a
high frequency of recoalescence events. If the emulsification is to be successful the formed droplets have to be protected. The repulsive interactions generated by the emulsifiers create a static protection.
The hydrodynamic interaction is crucial for the result of a collision due to
shear. The hydrodynamic interactions depend on the existence of an intenace
with an intenacial viscosity and elasticity. During the collision event, the intenace close to the approaching droplet is depleted of emulsifiers due to the
streaming ofliquid. The sunactant-depleted zone will then have a higher interfacial tension than the surrounding emulsifier-covered areas of the droplets.
This leads to sunace diffusion in the direction opposite to the liquid flow and
ensures the hydrodynamic resistance. If the emulsifier is oil-soluble, emulsi-

Physicochemical Aspects of an Emulsifier Functionality

167

fier from the internal part of the droplet will diffuse to the depleted area and
thereby reduce the hydrodynamic protection of the droplet.
The discussion in this section has been very qualitative, but an important
point is that the emulsifiers contribute to the emulsification as well as to the
stabilization. The role of the emulsifier for the stabilization is usually difficult
to identify in the simple type of shaking experiments that are the main background to the HLB, the PIT, and the phase diagram concepts. This type of simple, and thereby effficient, experiment provides information about both the
emulsifiability and the stability with a certain emulsifier.

6.7

The Emulsifier Surface

The ability of various food emulsifiers to generate adsorbed layers influencing


the interparticle interactions has been discussed. The type and magnitude depend on the composition of the surface generated from the adsorption process.
Foods usually are complex mixtures. They may contain both low molecular surface-active lipids and a versatile range of more or less surface-active proteins
and polysaccharides. The actual chemical composition of the emulsion droplet
surface is then the key factor that determines most of the surface interactions.
In systems containing several surface-active components, three types of adsorbed layers can be identified based on how the layers are formed. In reality,
the differences between the three adsorption structures discussed below are
not sharp, but this simplified description can provide a base when the properties of complex systems are discussed.

l. Competitive adsorption. A monolayer containing one predominant type of


molecule at the interface builds up through competition with other less
surface-active components that may be replaced in the interface.
2. Associative adsorption. An adsorbed layer containing a mixture of several
different surface-active components is formed.
3. Layer adsorption. One component adsorbs on top of the other.

6.7.1

Competitive Adsorption

In a system with several surface-active components, a homogeneous monolayer


is formed by the most surface-active component. The adsorption depends on the
main driving force for adsorbtion, mainly the hydrophobic interaction. Hence,

168

Food Emulsifiers and Their Applications

from a mixture of two emulsifiers, the most hydrophobic emulsifier will have the
strongest affinity to the interface. A consequence is that under competitive adsorption the component with the lowest water solubility will dominate the interface [e.g., the lowest critical micelle concentration (Kronberg, 1983)].
The character of the adsorbed layer, for instance its ability to generate repulsive interactions, is determined by the dominating compound. The structure of the layer depends on the geometrical shape of the molecules and on
lateral interactions between the molecules in the layer. Nonionic surfactants
may form very dense layers due to head-group attraction. Ionic surfactants are
able to form extremely loose layers due to inter-head-group repulsion.
An interesting experimental observation in agreement with this relation is
that the concentration of emulsifier necessary to obtain an emulsion is much
lower for ionic emulsifiers than for nonionic emulsifiers.
In a series of emulsions, we have studied the efficiency of the emulsification
(Ostberg et al., 1995) by droplet size measurements after homogenization. The
results show that for several emulsifiers very small droplets are obtained
(about 0.2 to 0.4 Jlm). The particle size obtained depends on the concentration
of emulsifier. The nonionic emulsifiers leads to a constant particle size down to
a critical concentration below which the ability to form emulsions is strongly
reduced. The critical concentration can be compared with the thickness of the
emulsifier layer on the emulsion droplet. The apparent thickness of the emulsifier layer can be estimated from the particle size and the concentration of
emulsifier (counted on the dispersed phase), if we assume that all emulsifier is
adsorbed to the interface. The apparent thickness gives the upper limit for the
adsorbed layer rather than the correct value:
Thickness of emulsifier layer =

0=

volume of emulsifier
emulsion droplet area
cern Vemulsion droplet

Aemulsion droplet

where cern is the emulsifier concentration (v/v) in the disperse phase.


The critical thickness (the thickness of the emulsifier layer at the critical
concentration) of the emulsifier layer can be compared with the size of the molecule. The results show a thickness of about 60% of the theoretical length of
the molecule for nonionic emulsifiers. Hydrophobic emulsifiers are less efficient during the emulsification and give very high values of the apparent thick-

Physicochemical Aspects of an Emulsifier Functionality

169

ness. The properties of the ionic emulsifiers are different. These emulsifiers
are able to emulsify the emulsions down to extremely low concentrations corresponding to very low surface concentrations (thin layers).

6.7.2 Associative Adsorption


In associative adsorption, a mixed surface is formed. The properties displayed
by the surface are some sort of average properties.
A typical associative system may be a long alcohol (for instance decanol)
and charged surfactants (for instance soaps). The alcohol acts as a spacer between the charged groups, which decreases the head-group repulsion within
the layer and reduces the surface energy. This increases the adsorption and
enhances the surface activity. Similarly, a lamellar phase is formed in the corresponding three-component phase diagram: water/sodium caprylate/deeanol
(Fontell, et al., 1968). Mixed layers are commonly formed due to associative
adsorption with natural and technical emulsifier blends. This is also a necessary requirement of the common assumption that an average HLB value should
describe the properties of an emulsifier blend (Davies, 1957). A common system assumed to act in this way is a mixture of sorbitan esters and ethoxylated
sorbitan esters where the smaller sorbitan esters can use the space between
the bulky ethoxylated esters (Boyd et al., 1976).
In the case of associative adsorption, both components are expected to be
present in the surface. If this situation is to be stable, the adsorption of the second component should be either enhanced by the presence of the first component or at least not influenced by it. The total amount of adsorbed material
should be greater than or equal to the sum of the two components.

6.7.3

Layered Adsorption

Adsorption in layers is possible when different classes of surface-active components are present in a mixture. See Table 6.9. The two components must be
very different in character to give a structure with a layered character rather
than a mixed layer. The second component adsorbs to a particle displaying the
characteristic properties of the primary adsorbing emulsifier. This usually
means a more hydrophilic surface, which can be expected to reduce the adsorbed amount. However, in some eases, the presence of certain groups increases the adsorption of specific substances.
The effects of the emulsifiers on protein adsorption is essential in most
emulsifier applications in the food industry.
Ethoxylated surfaetants usually give a strong reduction of protein adsorp-

170

Food Emulsifiers and Their Applications

Table 6.9

The apparent emulsifier layers for various emulsifiers estimated from


equation. (From Ostberg et al., 1995.)

Emulsifier
cone (%)1

Radius
(Jlm)2

Emulsifier-layer
thickness (1)3

Curve
shape 4

Estimated length
of the emulsifier

(1)5

Dodecylbenzenesulfate

0.1

0.47

1.6

15

Fatty acid monoethanolamid ethoxylate (7EO)

10

0.27

90

54

Fatty acid monoethanolamid ethoxylate (l3EO)

0.20

45

75

Fatty acid monoethanolamid ethoxylate (l8EO)

lO

0.23

59

93

The emulsifier concentration calculated on the oil phase.


The radius is shown as D(3, 2)/2.
3 The apparent emulsifier layer, estimated assuming that all emulsifier is estimated at the interface.
4 The curve shape shows the dependence for the apparent emulsifier layer of the emulsifier concentration.
5 The estimated length of the emulsifier molecule is estimated from the chemical formula or from
measurements in the corresponding lamellar phase.
1

tion. Courthaudon et al. (1991b) have shown C12E08 totally displace all adsorbed ~-casein from an emulsion system. Similar effects have also been obtained with emulsions formed with polysorbates (Dickinson and Tanai, 1992)
and with monoglycerides (Hall and Pethica, 1967). On the other hand, egg
yolk PC did not reduce the the adsorbed amount of ~-casein more than about
20% (Courthaudon et al., 1991a).
The adsorption of a range of plasma proteins at various phospholipid sur-

Physicochemical Aspects of an Emulsifier Functionality

171

faces has been characterized using ellipsometry (Malmsten, 1995). A large


variation of the adsorbed amount was obtained, depending on the combination
of protein and phospholipid. Purified PC and PE gave low adsorbed amounts,
while phosphatidic acid enhanced the adsorption of fibrinogen with a factor of
5 compared to a bare hydrophobic surface.

References
Arvidsson, G., et al. (1985). Eur. ]. Biochem, 152, 753-9.
Bancroft, W.D. (1913).]. Phys. Chem., 17, SOl.
Bergenstahl, B. (1991). In Food Polymers, Gels, and Colloids (ed. E. Dickinson), Royal
Society of Chemistry London, pp. 123-131.
- - - , Claesson, P. M. (1990). In Food Emulsions (eds. K. Larsson, S. Friberg),
Marcel Dekker, New York.
- - , Fontell K. (1983). Prog. Call. Pol. Sci., 68, p. 48-52.
- - , Stenius P.J. (1987). Phys. Chem., 91, 5944-48.
Boyd, J. V., et al. (1976). In Theory and Practice of Emulsion Technology (ed. A. L.
Smith), Academic, London.
Caffrey, M. (1985). Biochemistry, 24, 4826--44.
Courthaudon, J. L., et al. (1991a).]. Agr. Food Chem., 39, 1365.
- - et al. (1991b).]. Colloid Interface Sci., 145, 390.
Darling, D., Birkett, R. J. (1987). In Food Emulsions and Foams (ed. E. Dickinson),
Royal Society of Chemistry, London.
Davies, J. T. (1957). Proc. Intern. Congr. Surf. Activity, 2d, London, 1, 426.
Dickinson, E. (1986). Food Hydrocolloids, 1, 3.
- - - , Tanai S. (1992). Food Hydrocolloids, 6, 163-71.
- - et al. (1991). Food Hydrocolloids, 4, 403-14.
Eriksson, P. 0., et al. (1987). Phys. Chem., 91, 846-63.
- - et al. (1985). Chem. Phys. Lipids, 37, 357-71.
Fontell, K. (1978). Progr. Chem. Fats Other Lipids, 16, 145-62.
- - - et al. (1968). Acta Polytechnica Scandinavica, Chapter 2, Chemistry Series III,
74, 2.
Friberg, S. (1990). In Food Emulsions (eds. K. Larsson, S. Friberg), Marcel Dekker,
New York.
- - (1971).]. Colloid Interface Sci., 37,291.
--,Mandell, L. (1970a). ]. Assoc. Off. Chem. Soc., 47, 149.
- - (1970b). ]. Pharm. Sci., 59, 1001-4.
- - , Rydhag, L. (1971). Kolloid Z. u. Polymere, 244, 233-9.
- - - , Wilton, I. (1970). Liquid crystals-the formula for emulsions, Am. Parf. and
Cosm., 85, 27-30.
- - et al. (1969). ]. Colloid Interface Sci., 29, 155-6.
Gawrish, K., et al. (1992). Biochemistry, 31, 2856-64.
Griffin, W. C. (1979). In Kirk-Othmer Encyclopedia of Chemical Technology, Wiley, New
York, Vol. 8.
- - - (1949).]. Soc. Cosmetic Chemists, 311-26

172

Food Emulsifiers and Their Applications

Hall, D.G., Pethica, D.A. (1967). In Nonionic Surfactants (ed. M.J. Schick), Marcel
Dekker, New York, p. 516.
lnoko, Y., Mitsui, T.J. (1978). Phys Soc. ]ap., 44, 1918.
lsraelachvili, J. (1992). Intermolecular and Surface Forces, Academic, London.
--eta!. (1977). Biochim. Biophys. Acta, 470, 185-201.
---eta!. (1976). ]. Chem. Soc. Faraday Transactions II, 72, 1525.
Janiak, M.J., eta!. (1979). ]. Biol. Chem., 254, 6068-78.
Krog, N. (1990). In Food Emulsion (eds. K. Larsson, S. Friberg), Marcel Dekker, New
York, p. 127.
Kronberg, B. (1983). ]. Colloid Interface Sci., 96, 55-68.
Kunieda, H., Ishikawa, N. (1985). ]. Colloid Interface Sci., 107, 122-28.
- - , Shinoda, K. (1985). ]. Colloid Interface Sci., 107, 107-21.
Larsson, K., Krog, N. (1973). Chem. Phys. Lipids, 10, 177.
Lindblom, G., et al. (1991). Biochemistry, 30, 10938-48.
Malmsten, M. (1995). ]. Colloid Interface Sci., 172, 106--15.
Ostberg, G., et a!. (1995). Colloid Surfaces A, Physichemical Engineering Asp., 94,
161-71.
Pezron, 1., eta!. (1991).]. Colloid Interface Sci., 144, 449-57.
Rydhag, L. (1979). Fette Seifen Anstrichm., 81, 168-73.
--,Wilton, I. (1981). ]. Assoc. Off. Chem. Soc., 58, 830-7.
Shinoda, K., Friberg, S. (1986). Emulsions and Solubilization, Wiley, New York.
- - - , Kunieda H. (1983). In Encyclopedia of Emulsion Technology, Vol. 1 (ed. P
Becher), Marcel Dekker, New York.
---,Saito, H. (1968). ]. Colloid Interface Sci., 30, 258-63.
Small, D. M. (1986). Handbook of Lipid Research: Physical Chemistry of Lipids,
Plenum, New York.
Soderberg, I. (1990). Structural Properties of Monoglycerides, Phospholipids and Fats
in Aqueous Systems, PhD Thesis, University of Lund, Sweden.
Tanford, C. (1973). In The Hydrophobic Effect, Wiley, New York.
Walstra, P. (1983). In Encyclopedia of Emulsion Technology, Vol. 1 (ed. P. Becher),
Marcel Dekker, New York, p. 57.
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eta!.), IRL Press, Oxford, pp. 233-336.
Westesen, K., Wehler, T. (1992). ]. Pharm. Sci., 81, 777.
Wilton, 1., Friberg, S. (1971). Influence of temperature-induced phase transition in fat
emulsions,]. Assoc. Off. Chem. Soc., 48, 771-4.

SEVEN
Emulsifiers in Dairy Products
and Dairy Substitutes
Stephen R. Euston

7.1

Introduction

Bovine milk has been an important source of food for human beings for thousands of years. Not only is milk a very nutritious food in its own right, but it is
also a very versatile starting point for many other dairy products.
Milk is a complex food emulsion and colloidal sol. Table 7.1 gives the composition of whole cow's milk. The emulsion component is composed of fat droplets
dispersed in an aqueous phase containing protein. The protein is in the form of
both casein micelles, which are themselves colloidal particles, and free in solution as whey protein. A considerable reserve of knowledge has been assembled
on the structure and properties of the milk proteins (Swaisgood, 1992). The fat
droplets are stabilized by an adsorbed layer of protein and phospholipid called
the "milk fat globule membrane" (MFGM), which is distinct from the aqueous
phase protein (Walstra and Jenness, 1984). The average composition of the
MFGM has been estimated to be about 48% protein, 33% phospholipid, and

ll% water, with the remainder made up of other minor lipid components
(Walstra and Jenness, 1984). The phospholipid fraction of the membrane is composed of lecithin, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl
inositide, plasmalogens, and sphingomyelin. Phospholipids are important food

173

174

Food Emulsifiers and Their Applications

Table 7.1 Approximate composition of bovine milk. (From Walstra & Jenness, 1984,
Dairy Chemistry and Physics. Copyright 1984, Wiley-lnterscience. Reprinted by permission of John Wiley & Sons, Inc.)
Component

Average content
(wt%)

Water
Solids, nonfat
Fat in dry matter
Lactose
Fat
Protein
Casein
Mineral substances
Organic acids
Miscellaneous

87.3
8.8
32.0
4.6
3.9
3.25
2.6
0.65
0.18
0.14

Range (wt%)
85.5-88.7
7.9-10.0
21.(}.-38.0
3.8--5.3
2.4--5.5
2.3-4.4
1.7-3.5
0.53-0.80
0.13-0.22

Average dry
matter (%)
69.0
36.0
31.0
26.0
20.0
5.1
1.4
l.l

emulsifiers in their own right. The contribution that they make to the stability of
the milk fat globule is not well understood, but their use as food-grade emulsifiers has been the subject of extensive fundamental research (Courthaudon et al.,
1991; Dickinson et al., 1993a; Dickinson and Iveson 1993).
Until this century the conversion of milk to butter, cream, ice cream, and
various types of cheese had been more of a craft than a science. It is only relatively recently that an albeit incomplete understanding of these processes has
been available. It is now understood that the formation of these milk-based
products is a consequence of either the destabilization of the dispersed-phase
fat droplets (as in butter, ice cream, and whipped cream) or of the dispersed
aqueous-phase proteins (as in cheese). Emulsifiers play an important role in
the formation of structure in many dairy products.
In the following pages, the major emulsifier-containing dairy and imitation
dairy products will be reviewed. A brief description of their production will be
given where relevant, with emphasis on the role that emulsifiers play in the
formation and stability of the product.

7.2 Ice Cream


Ice cream is probably the most complex food that we encounter. It is both a
foam and an emulsion, and it contains ice crystals and an unfrozen aqueous

Emulsifiers in Dairy Products and Dairy Substitutes

175

phase whose freezing point is depressed by freeze concentration of salts, sugars, and polysaccharide stabilizers. Despite its obvious complexity, ice cream
has been widely studied, and much is known about the formation of its structure and the role that low molecular weight emulsifiers play in this (Covin and
Leeder, 1971; Lin and Leeder, 1974; Goff, 1988; Krog and Barfod, 1990;
Barfod et al., 1991). A typical ice cream formulation is shown in Table 7.2.

Table 7.2 A representative composition of ice cream.

(Based on Rosentahl, 1991, Milk and Dairy Products


Properties and Processing.)

Constituent

Weight percent in ice cream

Fat
Milk solid, nonfat
Sugar
Stabilizer
Emulsifier
Water

10.0
11.0
13.0
0.2
0.5
65.3

Ice cream manufacture is a relatively simple process. The ingredients are


mixed, heated to destroy pathogens, and then homogenized. The homogenization step is included to reduce the fat droplet size so that churning of the fat
does not occur upon whipping. An in-line pasteurization step is then carried
out prior to cooling to 4C and aging of the mix at this temperature for at least
4 hours. During this time milk proteins are able to redistribute between the fat
surface and the aqueous phase and fat crystallization occurs. The ice cream
mix is then aerated and frozen in a continuous freezer. Freezing is completed
by hardening the packed ice cream at minus 18C initially, and finally at minus 25C (Arbuckle, 1986; Rosentahl, 1991; Varnan and Sutherland, 1994).
An acceptable ice cream product can be made without the addition of low
molecular weight emulsifiers. It has been known for several years, however,
that incorporation of emulsifiers into ice cream results in a product that whips
more easily, is drier (a necessary requirement in molded products), has improved melt-down resistance, and has a smoother body and texture (Arbuckle,
1986). In addition to this, the ice cream has a higher overrun, the air is more
finely dispersed, and the foam structure is more stable if emulsifiers are present (Keeney, 1982). An understanding of the mechanism by which emulsifiers
change these properties has emerged over the past few years (Goff, 1988; Krog

176

Food Emulsifiers and Their Applications

and Barfod, 1990; Barfod et al., 1991). Before describing the role of emulsifiers in ice cream structure formation, it is pertinent to consider the structure
of the adsorbed layer formed around the fat droplet in the ice cream premix
emulsion. During the homogenization stage the ice cream mix is subject to
high shear. This results in the disruption of the fat phase into small droplets.
Surface-active components of the mix will adsorb onto the nascent-oiVwater
interface, lowering the interfacial tension and thus stabilizing the emulsion
droplets. The exact composition of the interfacial layer will depend on the proportions of each type of surface-active component and their relative surface
activities. Recent research has thrown light on the competitive adsorption between different milk proteins (Dickinson, 1986; Dickinson et al., 1988b;
Dickinson et al., 1989a; Dickinson et al., 1989c; Euston, 1989; Dickinson et
al, 1990b; Dickinson, 1992), milk proteins and emulsifiers (Dickinson and
Woskett, 1988a; Dickinson et al., 1990a; Dickinson, 1992; Euston et al.,
l995a; Euston et al., 1995b), and on the cooperative adsorption between proteins and polysaccharide stabilizers (Dickinson and Euston, 1990; Dickinson,
1992; Dickinson, 1993). These studies have identified that, in general, the
more surface-active component will predominate at the fat-droplet interface.
Thus the more surface-active protein in a mixture will dominate the adsorbed

layer initially, low molecular weight emulsifiers will, generally, displace protein from the surface with time, and under certain conditions polysaccharides
can interact with proteins and/or emulsifiers to contribute to the structure of
the adsorbed layer (Bergenstahl et al., 1992). The adsorbed layer of the ice
cream emulsion will be a composite of these functional ingredients.
The key to the formation of ice cream structure is the formation of a stable
foamed product. In ice cream this is achieved in two ways. The foam in ice
cream is not a typical protein-stabilized foam, where air bubble stabilization is
achieved by protein adsorption at the air/water interface. The initial stabilization of the foam network may indeed proceed by this mechanism, but prior to
freezing, the foam structure is stabilized primarily by the partial coalescence
of emulsion fat droplets at the air bubble interface. Figure 7.1 is a cryo-SEM
micrograph of ice cream. Adsorption of fat globules at the air bubble surface is
clearly visible. For this to be able to occur, the emulsion must be relatively unstable to the shearing forces exerted on the mix during air incorporation.
Several workers have attempted to demonstrate the link between emulsion coalescence stability and the mechanical or viscoelastic properties of the adsorbed protein layer (Doxastakis and Sherman, 1984; Rivas and Sherman,

Emulsifiers in Dairy Products and Dairy Substitutes

177

1984; Dickinson and Stainsby, 1988; Dickinson et al., 1988a). Dickinson et al.
(1988a) have shown a correlation between the surface shear viscosity of adsorbed layers of various proteins at the planar oil/water interface, and the coalescence stability of emulsions made with these proteins. The higher the
interfacial viscosity, the more stable the resultant emulsion under perikinetic
conditions. It has also been demonstrated that the orthokinetic stability of protein-stabilized emulsions (stability under turbulent or shearing conditions) is
reduced by the presence of low molecular weight emulsifiers (Chen et al.,
1993; Dickinson et al., 1993; Dickinson and Williams, 1994). The explanation
for this lies in the ability of the low molecular weight emulsifier to displace
protein from the fat-droplet smface, thus reducing the mechanical strength of
the adsorbed layer. Emulsifiers present at concentrations too low to cause significant protein displacement can interfere with interprotein interactions
within the adsorbed layer and reduce the interfacial viscosity in this way
(Dickinson et al., 1990a).

Figure 7.1 Cryo scanning electron micrograph of ice cream. Note the partial coverage
of the air bubble (AB) surface by fat globule (FG) and the partially coalesced fat globules (PC) in the foam lamellae. (Courtesy of A.B. McKenna.)

178

Food Emulsifiers and Their Applications

Emulsifiers that have an improving effect on the structure of ice cream do


so because they are able to aid in the destabilization of the milk protein-stabilized ice cream emulsion. Emulsifiers commonly used in ice cream mix such
as glycerol monostearate (GMS) and polysorbates, destabilize the emulsion by
displacing protein from the fat-droplet surface. This is a result of their greater
surface activity than milk proteins. Several studies have shown that the surface
activity of emulsifiers is strongly temperature-dependent. Studies on model
systems (Dickinson and Tanai, 1992) and in ice cream mix (Krog and Barfod,
1990; Barfod et al., 1991) indicate that displacement of protein from the
oiVwater interface by emulsifiers is at a maximum at temperatures between
4C and l0C. This observation provides an explanation for the improvement
in ice cream structure and stability imparted by the aging process. It is during
the aging step at 4C in ice cream manufacture that the displacement of the
majority of the protein occurs. Krog and Barfod and coworkers (Krog and
Barfod, 1990; Barfod et al., 1991) have investigated the aging effect in ice
cream emulsions and have shown that whereas protein displacement does occur during aging in the absence of emulsifiers, displacement is greater when
GMS is present, and greater still when glycerol monooleate has been added
(Figure 7.2).

The temperature dependence of emulsifier surface activity can be explained in terms of the phase behavior in aqueous solution. In the bulk phase,
monoglycerides exhibit a phase behavior similar to that of triglycerides (Krog,
1990). That is, they can exist in two polymorphic forms, the a. and ~forms,
that differ in the way the molecules pack in the crystal structure. When cooled
from a random molten state, a metastable crystalline structure termed the a.'
state is formed. Further cooling leads to a transition to the ~ state. The most
stable ~-crystalline structure will form if the a.' state is stored at ambient temperature. Lutton et al. (1969) proposed that the temperature dependence of
GMS surface activity can be explained by either the formation of a condensed
crystalline monolayer at the oiVwater interface at 5C or by micelle formation
close to the surface. The possibility that both mechanisms contribute to the increased surface activity on cooling cannot be ruled out.
Monoglycerides, and indeed other emulsifiers, have also been shown to exhibit a complex liquid-crystalline phase behavior in aqueous solution (Krog,
1990). Under certain conditions, the ~-crystal form of a monoglyceride will interact with water to form a lamellar liquid crystal. On cooling, this can transform to a so-called a.-gel phase (Krog, 1990). It has been hypothesized that

Emulsifiers in Dairy Products and Dairy Substitutes

179

30

Q)
rJ)

cc
..r::

c.

.....cc

20

c
c

Q)

0....

c.

eftQ)

>
;:;

10

cc

Q)

a:

24

Aging time at 5C

Figure 7.2 Changes in amount of protein adsorbed to fat globules in an ice cream mix
during the aging period at 5C. The amount of protein bound in the fat phase is calculated relative to the total protein in the mix. (From Krog, N., Barfod, N.M., AIChE
Symposium Series, 86, 1-6. Reproduced with permission of the American Institute of
Chemical Engineers. 1990 A!ChE. All rights reserved.)

phase transformations between different crystalline and/or liquid-crystalline


forms of the adsorbed monoglycerides are important in protein displacement
(Berger, 1990). During homogenization the temperature of the ice cream mix is
high (80C). Protein and emulsifier will both occupy the surface, with the
emulsifier having a relatively small effect on the protein surface coverage. As
the mix is cooled for aging, it is possible that a water-containing lamellar liquid-crystalline phase of the emulsifier is formed, which subsequently transforms into the a-gel phase. This latter transformation is accompanied by the
uptake of large quantities of water. At a later stage the more stable ~-crys
talline state may be formed. Berger (1990) believes that the two transformations, lamellar to a gel, and a gel to ~ crystal, will play a role in protein
displacement. The change from lamellar to a gel phase results in a decrease in
surface area of about 30%, and formation of the ~-crystal structure releases
large amounts of water (Berger, 1990). Darling and Birkett (1987), however,

180

Food Emulsifiers and Their Applications

believe that in ice cream insufficient emulsifier is present in the system for liquid-crystalline phases to form. For this to occur the emulsifier must be adsorbed on the fat droplet surface at a concentration far greater than that
required for monolayer surface coverage. This is not the case in ice cream
emulsions (Darling and Birkett, 1987).
Once the emulsion has been destabilized in the freezer, partial coalescence of the fat droplets has to occur at the air/water interface to partially
stabilize the foam structure prior to freezing. Research (Boode, 1992; Boode
and Walstra, 1993) has indicated that the crystal structure of the fat in emulsion droplets is important in determining their susceptibility to partial coalescence. Van Boekel (1980) has shown that when fat crystals form at the
surface of emulsion droplets, and are large enough to penetrate the adsorbed
layer, a lipid bridge can form between two droplets that are in contact with
each other. The proportion of solid fat in emulsion droplets is important in
determining instability (Walstra, 1987). If the majority of the fat is solid, coalescence, or partial coalescence will not occur, and the droplets will be stable. Similarly, if the droplets contain a very low proportion of solid fat,
coalescence can occur. If the solid fat content is in the approximate range 10
to 50%, partial coalescence is possible. A proportion of the emulsion droplet
is required to be in the form of liquid fat for partial coalescence to occur.
Walstra (1987) envisages a partial coagulum offat droplets as being held together by necks of liquid oil.
When fat crystallizes in dispersed emulsion droplets, considerable supercooling can be observed. Crystallization of fats occurs at nucleation points that
already exist in the fat phase. These nucleation points occur relatively infrequently in emulsion droplets where the fat is dispersed into a large number of
small droplets. Consequently, in dispersed systems the triglyceride needs to be
cooled below its bulk phase freezing point before crystallization is initiated.
Emulsifiers in adsorbed monolayers can act as templates for the surface crystallization of triglycerides. Emulsifiers containing saturated hydrocarbon
chains have been shown to be good initiators of fat crystallization, whereas
those with unsaturated hydrocarbon chains are not as good (Berger, 1990;
Barfod et al., 1991). Figure 7.3 gives the solid fat content (SFC) as a function
of time for model ice cream emulsions stored at 5C. Both saturated (GMS) and
unsaturated (GMO) emulsifiers initiate crystallization compared to control
emulsions with no emulsifier, but the SFC of GMS-containing emulsions is always greater than for those containing GMO.

Emulsifiers in Dairy Products and Dairy Substitutes

181

100
.....

c:

Q)
.....

c:

80

.....

.....co

60

0Cll

40

"0

20
0

24

Hours at 5C

Figure 7.3 Recrystallization of fat phase of ice cream emulsions without emulsifier
(control) or with saturated (GMS) or unsaturated (GMO) monoglycerides after cooling to
5C and aging. (From Barfod et al., 1991, and reproduced with permission.)

Darling and Birkett (1987) point out that in a mixed triglyceride system,
such as is found in milk fat, single discrete crystals are unlikely to form under
the rapid cooling conditions used in ice cream manufacture. They have shown
that in a cooled vegetable oil emulsion, concentric layers of triglyceride crystals are formed at the surface of oil droplets. These contain imperfections that
may be due to dislocations or recrystallization processes, and these are likely
to cause the destabilizing effect.
The ability of triglyceride crystals to penetrate the adsorbed layer depends
on a number of factors. The surface tension between crystal and oil, crystal
and water, and oil and water will determine how far into the oil phase the crystal will penetrate (i.e., if it is preferentially wetted by the oil or water phase).
Emulsifiers, by way of their surface activity, will alter these surface tensions,
and this may result in the crystals being able to penetrate further into the
aqueous phase. This would lead to a decrease in emulsion stability. The desorption of protein by emulsifier also aids destabilization by reducing the thickness of the layer through which fat crystals have to penetrate. The polymorphic
form of the fat crystal will also play a role in fat-droplet instability.
Triglycerides can exist in three general polymorphic forms, the a, ~. and W
polymorphs. When cooled from a melt, triglycerides will generally form a-type

182

Food Emulsifiers and Their Applications

crystals. These are not stable (Larrson and Dejmek, 1990) and will transform

into a Wpolymorph and subsequently to the stable ~ polymorph. Having been

formed at lower temperature, a crystals contain triglyceride in a more disordered liquid-like arrangement. These disordered crystals are softer and are
able to deform and follow the contours of the fat droplet more easily.
Consequently, they are less likely to penetrate the adsorbed layer. The ~-crys
talline structure is more solid-like, with the triglyceride molecules arranged in
ordered arrays. The ~ crystals have a greater mechanical strength and are unable to deform to the shape of the fat droplets. This leads to their bursting out
of the droplet into the aqueous phase (Darling, 1982).
In practice, two types of emulsifier are commonly used in ice cream: monoand diglycerides and polyoxyethylene derivatives of glycol or glycol esters, for
example polysorbates (Keeney, 1982). Sucrose esters have also been evaluated
and have been found to be suitable as ice cream emulsifiers (Buck et al.,
1986). Mono- and diglycerides and polysorbates are usually all found in current ice cream emulsifier blends. The explanation for this lies in the relative
abilities of polysorbates and mono- and diglycerides as emulsion destabilizers,
or as foam-forming agents. Polysorbates are far more efficient at displacing
protein from the oiVwater interface than are mono- and diglycerides and thus
are better emulsion destabilizers (Keeney, 1982). Mono- and diglycerides are
better foaming agents and thus are able to aid the formation of the initial foam
prior to fat-droplet agglomeration at the air/water interface (Keeney, 1982). A
second factor is the differing abilities of emulsifiers to influence fat crystallization. Figures 7.2 and 7.3 show that whereas GMO is able to displace more protein from the fat globule surface during aging than does GMS, GMS initiates
more fat crystallization than GMO. Use of a mixed emulsifier system would
also allow optimum protein displacement combined with optimum fat crystallization.
In summary, nonprotein emulsifiers are important in ice cream in several
respects:

l. They promote protein desorption from the surface of fat droplets, both by
their higher relative surface activity and the possible formation of liquid-crystalline mesophases.
2. They can act as nucleation points for surface crystallization of triglycerides.
3. They may promote fat crystal penetration of the adsorbed protein layer by
alteration of the surface tension between various phases.

Emulsifiers in Dairy Products and Dairy Substitutes

183

4. They may help in the initial formation and stabilization of the ice cream
foam prior to partial fat-globule coalescence and freezing. Monoglycerides
are particularly good at this function.

7.3 Whipped Cream and Whipping Cream


The distinction between whipped cream and whipping cream is one of colloidal
state. Whipping cream is an oil-in-water emulsion stabilized by adsorbed milk
protein and (where added) low molecular weight emulsifier. Whipping cream can
be made by concentration of the milk fat globules found naturally in milk, or by
a recombination process where amorphous milk fat is homogenized with milk
proteins and emulsifiers. The fat content of whipping cream is about 35% by
weight. Unlike ice cream emulsion, which only has to be stable enough to be
aged for a few hours before processing into ice cream, whipping cream emulsion
has to be stable enough to allow storage for several weeks at ambient temperatures, if UHT processed, without appreciable loss of stability.
Whipped cream is formed from whipping cream when air is incorporated
into the emulsion to form a foam. The structure of whipped cream resembles
that of ice cream in some ways. The foam is stabilized, initially by adsorbed
protein and any added emulsifier. Prolonged whipping of the cream leads to
partial agglomeration of fat globules at the air/water interface. Whereas in ice
cream the final structure is partially stabilized by fat-globule adsorption at the
air bubble surface, but mostly by freezing of the aqueous phase, in whipped
cream the higher dispersed-phase fat content (35 wt% compared to about 10
wt% in ice cream) leads to a higher degree of fat-particle coalescence at the
air/water interface. This greater fat adsorption leads to formation of a stable
foam without the need for freezing. Figure 7.4 is a cryo-SEM micrograph showing fat-globule adsorption at the air/water interface in whipped cream.
Comparing this to Figure 7.1, a cryo-SEM micrograph of ice cream, it is apparent that structural similarities exist, but that the degree of fat-globule adsorption appears to be less in ice cream. The adsorbed fat globules contribute to
the rheological properties of the foam. By influencing drainage in the aqueous
lamellae between air bubbles, the partially coalesced, adsorbed fat globules
impart a small but finite yield stress on the whipped product (Dickinson and
Stainsby, 1982). This allows whipped cream to "stand up" under its own
weight even at ambient temperature.

184

Food Emulsifiers and Their Applications

Figure 7.4 Cryo scanning electron micrograph of whipped cream. Note the greater coverage of the air bubble (AB) surface by adsorbed fat globules (FG) than occurs in ice
cream (Figure 7.1). (Courtesy ofA.B. McKenna.)

Nonhomogenized cream separated from milk will whip satisfactorily without addition of emulsifiers. If the cream is homogenized prior to whipping and
the mean fat-globule size is reduced, emulsifiers have to be added to aid destabilization. Anderson and Brooker (1988) have attributed the differences in
whipping ability of homogenized and nonhomogenized cream to the differences in interfacial composition of the emulsion droplets in these systems.
Nonhomogenized cream contains fat droplets stabilized by native MFGM. The
composition of this has been described earlier (Section 7.1). After homogenization, the particle size is reduced, interfacial area is increased, and consequently, native MFGM is insufficient on its own to stabilize the newly formed
interface. A combination of increased interfacial area and competitive adsorption between MFGM material and cream serum proteins means that proteins
from the aqueous phase (caseins and whey proteins) contribute to the interfa-

Emulsifiers in Dairy Products and Dairy Substitutes

185

ciallayer. The adsorbed layer in homogenized cream has been found to consist
mainly of caseins, with smaller amounts of ~-lactoglobulin and ~-lactalbumin
(Anderson et al., 1977; Darling and Butcher, 1978; McPherson et al., 1984).
This is consistent with the available data on competitive adsorption between
casein and whey proteins in model systems (Euston, 1989; Dickinson et al.,
1990b). In the early stages of whipping, before fat-globule adsorption and partial coalescence occurs to any great extent, the air bubbles are stabilized by
adsorbed milk serum proteins. Since this air/water interface is composed of the
same proteins that surround the fat/water interface in homogenized cream, the
difference in interfacial tension between the two interfaces is not very great
(Anderson and Brooker, 1988). Interfacial tension differences have already
been put forward as a driving force for fat-globule adsorption at the air/water
interface in ice cream (Section 7.2). Because of the small interfacial tension
differences between fat globule and air bubble in homogenized cream, the driving force for fat-globule adsorption is low. In nonhomogenized milk the interfacial tension differences between a fat globule stabilized by MFGM and an air
bubble stabilized by cream serum proteins are sufficient to act as the driving
force for fat-globule adsorption at the surface of air bubbles. Of course, this is
also aided by the presence of fat crystals at the fat-globule surface and by the
shearing forces introduced during whipping.
The importance of the fat phase manifests itself in two ways. As in ice
cream (Section 7.2), fat crystals are known to be important in the shear-induced coalescence of the fat globules (Darling, 1982), and the presence of a
certain amount of liquid fat is a prerequisite for good whipping properties.
Bucheim (1986) put forward the idea that the interfacial layer surrounding the
fat droplets ruptures when they collide during agitation. The subsequent
spreading of liquid fat is the first stage in destabilization by aggregation of adjacent droplets. The importance of the solid fat content of the fat globules has
been demonstrated by Darling (1982), who observes a direct correlation between the SFC and whipping time in natural cream.
Recombining technologies are becoming an increasingly important process
for making whipping cream bases. These would encounter the same problems
as homogenized cream (i.e., the similarity in composition and interfacial tension between air/water and oil/water interfaces), if they were formulated with
milk protein as the only surface-active material. For this reason, recombined
whipping creams, and indeed homogenized natural creams contain added low
molecular weight emulsifiers. These will alter the composition of the fat-

186

Food Emulsifiers and Their Applications

droplet surface, thus changing the interfacial tension. Addition of an optimum


concentration of emulsifiers results in an oil/water interface composed of protein and emulsifier differing sufficiently from the air bubble interface for fatglobule adsorption to occur on whipping. For similar reasons to those proposed
for ice cream (Section 7.2), whipping cream emulsifiers are usually a combination of two types. There is often a lipophilic emulsifier such as GMS, or one of
its derivatives, and a water-soluble polyoxyethylene derivative such as one of
the Tweens. Thome and Eriksson (1973) have shown that the amphoteric phospholipids are good emulsifiers in whippable emulsions when used in combination with monoglycerides. This is a significant observation when the trend
toward natural, nonsynthetic emulsifiers is considered. Phospholipids are a
natural component of milk fat and can be produced as a byproduct when milk
is processed into, for example, butter. This can explain the increasing use of
buttermilk powders as combined emulsifier/protein systems in whipped emulsions (Vodickova and Forman, 1984).
The structure of the composite fat-globule surface layer in homogenized
and recombined dairy whipping cream is not known for certain. Two theories
have been put forward. One theory (Krog, 1977) maintains that a primary layer
of emulsifier adsorbs at the fat-droplet surface, and that a secondary layer of
protein is attached to this primary layer through relatively weak cooperative
hydrogen bonding. When the cream is whipped this protein layer is removed
from the fat globule relatively easily, and the emulsion destabilized in this way.
Doxastakis and Sherman {1984), however, have evidence that the protein and
emulsifier form a mixed interface where both are adsorbed through hydrophobic interaction with the interface. It is speculated that this can lead to localized differences in the interfacial tension at the fat-droplet surface (i.e.,
between protein-rich and emulsifier-rich regions of the adsorbed layer), which
helps to drive fat-globule adsorption and partial coalescence. Whichever of
these theories is correct, the elements are also relevant to the destabilization of
ice cream emulsion when it is frozen and whipped.
A stable whipped cream can be formed from whipping cream that has been
held at room temperature for some time. However, a superior product is obtained if the cream is aged for several hours at low temperature prior to whipping. This, as in ice cream, is a consequence of increased emulsifier surface
activity at low temperature.
It would appear, therefore, that the functions of emulsifiers in whipping
cream are essentially the same as for ice cream, i.e.,

Emulsifiers in Dairy Products and Dairy Substitutes

187

l. They destabilize the cream through their ability to displace protein from
the oil/water interface. This changes the adsorbed layer composition
and interfacial tension of the fat droplet.
2. They may destabilize the emulsion through their ability to form lyotropic
liquid-crystalline mesophases and the subsequent phase transformations that occur to form stable crystalline forms.
3. They may participate in the initial foam stabilization.
4. They aid in the formation of fat crystals at the fat-droplet surface, which
crystals are essential for fat-globule partial coalescence.
This last point is reinforced by electron microscopic studies of emulsifier
containing whipping creams. Figure 7.5 is a cryo-TEM micrograph of a whipping cream containing Tween 80. The large needle-like crystals that emanate
from the fat droplet are, apparently, typical of those formed in Tween-containing systems (McKenna, 1995 personal communication). The bridging of two fat
droplets by these needle-like crystals is clearly visible.

7.4 Whipped Toppings


Whipped toppings, usually made with hydrogenated (hardened) vegetable oils,
have been a popular dairy analog product for several years. Table 7.3 gives a
typical composition for a whipped topping powder.
Si (1991) lists the types of emulsifier used in whipped toppings as propylene glycol esters of monoglycerides, acetic acid esters of monoglycerides, or
lactic acid esters of monoglycerides.
In whipped toppings, as in dairy whipping creams, the emulsifiers appear to

Table 7.3 Typical whipped topping powder


composition. (From Si, 1991. Reprinted by permission of the Society of Dairy Technology.)
Ingredient

Composition (%)

Hardened coconut or
palm kernel oil
(melting point 3l-36C)
Maltodextrin
Sodium caseinate
Emulsifiers

52.0

32.0
8.0
8.0

188

Food Emulsifiers and Their Applications

Figure 7.5 Cryo transmission electron micrograph of whipped cream containing Tween
80 as emulsifier. A partial agglomerate of fat globules is shown. The spikes emanating
from the fat globule (FG) are fat crystals (FC) and can clearly be seen to bridge between
two fat droplets (B). These spikes are, apparently, features of Tween-containing systems. (Courtesy of A.B. McKenna.)

be important in the destabilization of the emulsion, while the protein is important in giving initial stability to the oil-in-water emulsion. The mechanism by
which emulsion destabilization is achieved, however, is different. Whereas
whipped dairy creams and whipped imitation creams are stabilized by partially coalesced, relatively intact fat globules adsorbed at the air/water interface, whipped toppings are stabilized by crystalline fat at the air bubble
surface. Krog and coworkers (Barfod and Krog, 1987; Bucheim et al., 1985;
Krog et al., 1986) have carried out extensive studies of the factors that effect
structure formation in whipped toppings. They have shown (Barfod and Krog,
1987) that part of the fat in spray-dried topping powders is in a supercooled
state. When these topping powders are reconstituted in water at low temperature, they show large structural changes that determine whipping characteris-

Emulsifiers in Dairy Products and Dairy Substitutes

189

tics and foam structure. The emulsion becomes unstable due to spontaneous
recrystallization of the supercooled fat. The destabilization of the emulsion is
probably promoted by the temperature-dependent desorption of protein from
the surface, followed by coalescence. This makes crystallization of the supercooled fat more likely, due to the increased probability of nucleation sites
(Bucheim et al., 198S).
Scanning electron microscopy studies (Bucheim et al., 198S) show that the
final structure of the aerated whipped topping is stabilized by a layer of crystalline fat of about 0.1jlm thickness. The aqueous phase lamellae between air
bubbles also contains large proportions of crystalline fat, with smaller proportions of relatively intact fat globules.
The kinetics of fat crystallization and emulsion destabilization depend on
the type of emulsifier used in the formulation. Bucheim et al. (198S) have investigated the effect of distilled propylene glycol monostearate (PGMS), distilled unsaturated monoglycerides (glycerol monooleate, GMO), and distilled
saturated monoglycerides (glycerol monostearate, GMS) on the structure of
whipped toppings. Only PGMS is typically used in commercial formulations.
Figure 7.6 shows that all the emulsifiers promote fat crystallization when compared to toppings without added emulsifier. The effect of PGMS and GMO,
however, was greater than for GMS. The increased emulsion destabilization
caused by the enhanced fat recrystallization led to PGMS- and GMO-containing whipped toppings being more stable than those made from GMS-containing emulsions (Bucheim et al., 198S). The GMS-containing reconstituted
topping emulsion is too stable to allow consequent stabilization of incorporated
air (whipping).
The importance of protein desorption on the whipped topping structure and
stability has been demonstrated by Krog and coworkers (Krog et al. 1986;
Barfod and Krog, 1987). Table 7.4 gives the percentage protein contents of the

soc and
soc in the ab-

fat and aqueous phases of whipped topping powders reconstituted at


30C, and containing PGMS, GMS, or no added emulsifier. At

sence of emulsifier, almost one-quarter of the protein is associated with the fat
phase. This falls to 1.3% when PGMS is present and 7.7% with GMS added.
The temperature dependence of protein displacement is also evident. At 30C,
over 40% of the protein is in the fat phase when emulsifier is absent, but this
drops to about 33.7% in the presence of PGMS and 12.3% when GMS is included in the formulation (Krog et al., 1986). The GMS-containing reconstituted toppings have a significant proportion of protein still associated with the

190

Food Emulsifiers and Their Applications

40

~
.....
c
Q)
.....
c
0

.....Ctl

30

......

"0

(/)

20

...

...

...

10
0

10

20

30

Time (min) at 15C

Figure 7.6 Crystallization of supercooled lipid fractions in topping emulsions with different surfactants, reconstituted (I :3) in deuterated water (D20), measured by pulsedNMR at l5C. e GMO (Dimodan 0), PGMS (Promodan SP), o GMS (Dimodan PV),
A no surfactant added. [From Bucheim et al. (1985). Reprinted by permission of
Scanning Microscopy International.]

Table 7.4 Distribution of protein between the fat cream phase and the water

phase of centrifuged topping emulsion. (From Barfod and Krog, 1987. Reprinted by
permission of the American Oil Chemists Society.)

Surfactant

After I hour
at SC
%protein in
fat phase

After I hour
at SC
%protein in
water phase

After I hour
at 30C
%protein in
fat phase

After I hour
at 30C
%protein in
water phase

l0%PGMS
l0%GMS
None

1.3
7.7
24.0

98.7
92.3
76.0

33.7
12.3
41.7

66.3
87.7
58.3

fat-droplet surface, even after aging of the emulsions at 5C. This is obviously
enough to form an adsorbed layer strong enough, in combination with the lower
degree of fat recrystallization, to prevent stabilization of the incorporated air
bubbles. This is in contrast to the situation in ice cream and whipping cream,
where GMS is capable of destabilizing the fat emulsions to a degree that partial coalescence can occur. Darling and Birkett (1987) point out that the level

Emulsifiers in Dairy Products and Dairy Substitutes

191

of emulsifiers in whipped toppings is sufficient to allow the formation of emulsifier adsorbed layers of far greater than monolayer coverage. They suggest that
the formation of liquid-crystalline mesophases, and the a.-gel phase of the adsorbed emulsifier, is a possibility. Westerbeek et al. (1991) have shown that a
common emulsifier used in whipped toppings, glycerollactopalmitate (GLP),
is capable of forming the a.-gel phase at oil/water interfaces, and this may contribute to emulsion destabilization.
The present understanding of the formation of structure in whipped toppings suggests the following mechanism for emulsion destabilization and formation of the foam structure:
1. The powdered, dried topping emulsion is stable.
2. When reconstituted at low temperature, protein desorption is initiated and
is aided by added emulsifiers such as PGMS. The mechanism of protein
desorption will be similar to that already described for ice cream. There is

an almost total desorption of protein from the fat droplet surface.


3. Coalescence of fat droplets can occur during whipping, due to the presence of a liquid fat produced by supercooling.
4. Coalescence leads to a recrystallization of the supercooled fat, which is
again aided by the presence of emulsifiers.
5. A continuous phase of elongated fat crystals is formed, resulting in an increased viscosity of the whip, which is capable of stabilizing dispersed
air bubbles.
As in ice cream, the functions of emulsifiers in whipped toppings are to promote protein desorption and fat crystallization.

7.5

Cream Liqueurs

Cream liqueurs are dairy emulsions of high added value. The combination of
milk protein-stabilized cream emulsion and high alcohol content make cream
liqueurs unique among dairy emulsions. Table 7.5 gives a typical range of
compositions for cream liqueur. In practice many commercial formulations
also have small amounts of GMS added.
The production of cream liqueurs is governed by the relative poorness of
the alcoholic aqueous phase as a solvent for proteins and sugars. Two commercial processes are in common use (Banks and Muir, 1988), namely, the single-

192

Food Emulsifiers and Their Applications

Table 7.5 Range of compositions of a standard cream liqueur. (Reprinted from Banks

et al., 1981, with permission.)

Component

Composition (wt%)

Milk fat
Added sugars
Sodium caseinate
Nonfat milk solids
Ethanol
Water

12-16
15-20
2.6-3.5
l.0-3.5
14
46-51

stage process and the two-stage process. Figure 7. 7 presents flowcharts for
both processes. The main difference between the two processes lies in the
stage at which the alcohol is added. In the single-stage process this is prior to
homogenization, whereas in the two-stage process it is after homogenization.
Banks and Muir (1988) found that homogenization in the presence of alcohol
leads to the formation of fewer large fat globules, and as such is preferable in
terms of emulsion stability. A characteristic of cream liqueur production is the
harsh homogenization conditions used (two passes at 300 bar). This results in
a product in which more than 97% of the fat droplets have a diameter less than
0.8 f..Lm. A second factor favoring formation of smaller fat droplets is the significant lowering of interfacial tension observed at the oil/water interface when
alcohol is added to the aqueous phase (Bullin et al., 1988; Dickinson and
Woskett, 1988; Burgaud and Dickinson, 1990). As a result of the very fine
droplet size, the protein in the added cream has to be supplemented by sodium
caseinate (to a fat-to-caseinate ratio of approximately 5:1) to provide adequate
coverage of the newly formed fat surface by protein (Banks et al., 1981). The
fine particle size of the dispersed fat droplets gives the product an excellent
stability with respect to creaming. Banks et al. (1981) have noted no signs of
creaming in liqueurs with a composition within the range quoted in Table 7.5
after 12 months storage. The high level of added sodium caseinate, however,
leads to cream liqueur emulsions being unstable in acid environments. This
means that they are not suitable for combination with acidic beverage mixers
such as lemonade. A cream liqueur that is stable in an acid environment can
be made by replacing the sodium caseinate with GMS. The emulsifier replaces
milk protein as the primary emulsion stabilizer at the oil/water interface, and
the nonadsorbed protein is unable to aggregate the fat droplets when exposed

Emulsifiers in Dairy Products and Dairy Substitutes

193

Single-Stage Process
Water

Sugar

Caseinate

Cream

Cream Base

Citrate

Alcohol

Product

Homogenize
2 x 300 bar, 55"C

Two-Stage Process

Cream Base
Homogenize
2 x 300 bar, 55"C
coolto<21rC

Homogenized Base
Gentle mixing

Alcohol
Product

Figure 7.7 Flow diagrams for the process of manufacture of a cream liqueur in (a) a
single stage and (b) two stages. (From Banks and Muir, 1988. Reprinted by permission

of Elsevier Applied Science Publishers.)

to acidic surroundings (Banks and Muir, 1988). Acid stability in this type of
product is gained at the expense of emulsion stability and shelf life. In practice, legal limits in some countries set the concentration of GMS at no more
than 0.4 wt%, and so total replacement of caseinate by GMS is not feasible.
Many manufacturers add low concentrations of GMS as well as sodium caseinate to cream liqueur formulations. Dickinson and coworkers (1989b) have
shown that, in addition to displacing some, but not all of the milk protein from
the fat droplet surface, which presumably infers some acid stability on the
product, GMS also improves the creaming stability of a model cream liqueur.
When model cream liqueurs were stored at room temperature for 12 weeks, no
creaming was observed with added GMS concentrations above 0.5 wt%. Below
this level of added GMS a reduced degree of creaming was observed compared
to control samples with no emulsifier (Dickinson et al., 1989b). The increased
creaming stability was associated with rheological changes in the emulsifier
aqueous phase. At low CMS concentrations the emulsions exhibit Newtonian
behavior, whereas above 0.5 wt%, a clear yield stress is found. Dickinson et al.

194

Food Emulsifiers and Their Applications

(1989b) postulate the formation of a weak gel-like network in the continuous


phase formed by interaction of caseinate with GMS. It is also likely that interaction between caseinate and GMS at the oil/water interface plays a role in the
creaming stability. Evidence for interactions between adsorbed caseinate layers and GMS has been reported by Doxastakis and Sherman (1984), who investigated the surface rheological properties of mixed caseinate GMS systems.
An apparent contradiction in the work of Dickinson et al. (1989) is that although creaming stability is enhanced at GMS levels above 0.5 wt%, the shelf
life, as tested using an accelerated method at 45C, decreases in this region
(Figure 7.8). Dickinson et al. (1989) point out that whereas weak gels are able
to prevent formation of a substantial cream layer, they are also prone to slow

150 . ------------------------------,

GMS concentration (wt%)

Figure 7.8 Effect of GMS on the shelf life of simulated cream liqueurs on storage at
45C. The time for serum separation to first become visible is plotted against the GMS
concentration. Different symbols refer to separate experiments.(Based on Dickinson et
al., 1989. Reprinted by permission of the Institute of Food Technologists.)

Emulsifiers in Dairy Products and Dairy Substitutes

195

syneresis when stored for any length of time. This leads to separation of the
aqueous phase and formation of a distinct, clear serum layer at the bottom of
the sample container. Whether this syneresis will occur at room temperature is
not certain, and Dickinson et al. (1989b) stress that a correlation between the
shelf life at 45C and that at room temperature may not follow. Cream liqueurs
stored under ambient conditions can have shelf lives of several years. Clearly
these are likely to be consumed before serum separation becomes evident.
Since the legal limits on the amount of emulsifier that can be added are set at
about 0.4 wt%, the problem of gel syneresis is unlikely to be encountered. At a
level of 0.4 wt% added GMS, creaming under gravity would not be eliminated
completely, but would be reduced to a level acceptable to the consumer
(Dickinson et al., 1989b).

7.6

Creams and Coffee Whiteners

Cream products containing 10 to 20% fat have been popular as coffee creamers for over 50 years (Abrahamson et al., 1988). Coffee creamers and whiteners
perform several functions: they give coffee a white color, reduce bitter taste by
complexation of the tannic acids with milk proteins, give the coffee a creamlike flavor, and give body to the coffee (Sims, 1989).
Traditionally, coffee cream is produced by simple concentration of milk up
to the required fat content. The cream is usually heat treated using a UHT
process, homogenized either before or after heating, and packed aseptically to
give a long shelf life. Emulsifiers are not usually added to this product. More
recently, with the advent of recombining technology and with the preference of
some consumers for vegetable oil-based products over those containing milk
fat, new products have appeared that require the addition of emulsifiers to
their formulations. If a recombined coffee cream is produced, the formulation
is more complex than for coffee cream. Table 7.6 gives a typical formulation
for a recombined coffee cream containing 19% milk fat, as suggested by
Zadow (1982).
The recombining process involves a two-stage homogenization with an 18MPa first stage and a 3-to-4-MPa second stage. Presumably the emulsifiers are
added to aid in the homogenization process by reducing the energy required to
form the fat/water interface. It may also be assumed that since in natural coffee
creams the milk fat is already in a dispersed state, the energy required to reduce the particle size is less, and so emulsifiers are not needed. However,

196

Food Emulsifiers and Their Applications

Table 7.6 Typical formulation for a recombined coffee cream. (From Zadow, 1982. Reprinted by permission of the International Dairy
Federation.)

Ingredient

Composition (wt%)

Skim milk powder


Buttermilk powder
Anhydrous milk fat
Carrageenan
GMS
Tween 60
Water

3.0
4.5
19.0
0.03
0.05
0.1
73.32

Zadow (1982) states that emulsifiers and stabilizers are required only if the
product is to be given a high heat treatment (a UHT process or steam injection). This may indicate a role for the emulsifiers in protecting the protein or
emulsified fat from heat damage. Evidence exists to support this hypothesis
and will be dealt with in more detail in Section 7.8.
Whereas coffee cream and recombined cream are used in a liquid form, coffee whiteners based on vegetable fat are also popular in dry powder form. Typical
formulations for liquid and dry powder coffee whiteners are given in Table 7.7.
Table 7.7 Typical formulations for liquid and powdered coffee whiteners. (From Si, 1991. Reprinted by permission of the
Society of Dairy Technology.)

Ingredient

Composition (wt%)
Liqnid

Powder

Fat

10.0

Sodium caseinate
Maltodextrin (DE28)
Monoglycerides
Tartaric acid esters
of monoglycerides
Carrageenan
Sodium alginate
K2HP04
Flavor
Water

l.O

30.0
4.0
62.0
-1.5
0.5

10.0
0.2
0.2
0.05
0.2
300 ppm
to 100%

0.05
1.5
lOOOppm

Emulsifiers in Dairy Products and Dairy Substitutes

197

Other water-soluble surfactants, such as Tween 60, are commonly used in


place of the tartaric acid esters of monoglycerides. Si (1991) states that the function of the emulsifiers in coffee whitener is to improve whitening ability and to
aid powder dispersibility in coffee. Knightly (1969) has found that GMS is more
effective in improving powder dispersibility, whereas Tween 60 is better at improving the rate of solution of the powder. Optimum whitening ability is attributed to small fat globules and a narrow particle-size range, and its attainment in
coffee whitener has been attributed to the presence of the GMS and its derivatives (Si, 1991). Whitening power in a dispersion is related to the surface area of
the disperse particles. The higher the surface area the greater the light reflectance from the dispersion and thus the greater the whitening effect. This is
true for both dairy coffee creams and nondairy coffee whiteners. Leo and
Betscher (1971) have noted that there is an optimum particle-size range for optimum optical density and whitening power of the dispersion. Overhomogenization
of a coffee whitener formulation is known to result in a loss of whitening power.
The emulsifiers added to powdered formulations prior to spray drying are
capable of stabilizing the emulsion in the liquid form. Sodium caseinate is
usually required to give stable fat droplets in the dried powder (Sims, 1989),
since an adsorbed proteinaceous layer is better able to withstand the extreme
conditions in the drier. Because sodium caseinate is used at high concentration (typically in the range 3 to 15%), ways of reducing the amount in coffee
whitener have been sought. One way of doing this is to use sodium (or calcium)
stearyl lactylate or sodium stearyl fumarate as an emulsifier. Miller and
Werstak (1983) have used 2.5% monoglycerides plus sodium stearyl-2-lactylate (SSL) in the approximate ratio of 7.3:1. They claim a reduction of sodium
caseinate to 60% of that required in normal formulations. The function of SSL
appears to be through its ability to form a complex with sodium caseinate (Leo
and Betscher, 1971). It is likely that this interaction results in improved fat encapsulation in the dried state through increased interfacial rigidity of the adsorbed layer. This is analogous to the increased emulsion coalescence stability
observed when GMS complexes with protein adsorbed at the oil/water-emulsion interface (Doxastakis and Sherman, 1984; Rivas and Sherman, 1984).

7.7

Processed Cheese

Processed cheese manufacture dates back approximately 100 years.


Originally, it was used as a way of increasing the shelf life of cheese and of uti-

198

Food Emulsifiers and Their Applications

lizing lower quality cheese (Carie et al., 1985). To manufacture processed


cheese, the cheese raw material (a mixture of rennet and fresh cheeses) is first
cleaned, chopped, and heated at 70 to 82C with emulsifying salts and other
additives. Heating and water addition are often combined by using direct
steam injection. The pH of the mix is lowered to 5.6 to 5.8 using organic acids
such as citric or lactic, and the product is then extruded into packages
(Rosenthal, 1991). Alternatively, the correct pH can be obtained by careful selection of a blend of polyphosphate emulsifying salts, which have some buffering capacity in this application (Lee, S.K., 1995 personal communication). The
final product can have 15 to 25% fat and up to 58% water.
The structure of processed cheese is one of fat droplets dispersed in a concentrated, gelled protein network. Emulsion stability in the fat droplets is controlled,
primarily, by adsorbed caseins or hydrolyzed casein fractions. Some manufacturers add mono- and diglycerides as emulsifiers. The structure and texture of
processed cheese is closely linked with the size and distribution of fat globules in
the cheese (Thomas et al., 1980; Shimp, 1985).lf the fat in a processed cheese is
weakly homogenized and large fat droplets are formed, the cheese is soft and
melts easily. lf the fat droplets are small, the cheese is hard and nonmelting.
To control the structure of processed cheese, so-called emulsifying salts such
as polyphosphates are added. Although these are not surface active they play an
important role in modifying the emulsifying activity of the surface-active caseins. Caseins bind calcium, and this has the effect of reducing their solubility,
and thus their emulsifying activity. Through their ability to chelate calcium more
strongly than the caseins, emulsifying salts are able to improve the solubility and
emulsifying properties of the caseins. Emulsifying salts are of two types: those
that bind calcium relatively weakly and those that bind calcium more strongly.
Weak emulsifying salts have a modest effect on the emulsifying properties of the
caseins and lead to the formation of a soft cheese with relatively large fat
droplets. Strong emulsifying salts give a greater improvement in the emulsifying
capacity and result in a hard cheese with smaller fat droplets.
The use of low molecular weight, surface-active emulsifiers (Tweens and
Spans) was first investigated in the 1950s (Holtorff et al., 1951). They are not
as good as emulsifying salts in promoting structure formation in processed
cheese, and in some cases they act to destabilize the fat emulsion by protein
displacement from the surface.
Concern has been expressed over the nonnutritional effect of forming a
phosphorus/calcium complex. The supplementation of emulsifying salts by

Emulsifiers in Dairy Products and Dairy Substitutes

199

monoglycerides has been investigated as a way of reducing the concentration


of emulsifying salts. Gavrilova (1976) produced processed cheese of improved
rheology and shelf life using an emulsifying salt/monoglyceride mixture.
Zakhorova et al. (1979a, b) achieved a 50% reduction in the concentration of
emulsifying salts required by adding 1% monoglyceride to the cheese. The
processed cheese produced was reported to be of good quality and to have improved hydrophilic properties.
Lee et al. (1995) have studied the effect of adding small concentrations of
low molecular weight surfactants as coemulsifiers in combination with emulsifying salts in a model processed cheese. The surfactants used were sodium dodecyl sulphate (SDS, an anionic surfactant), cetyl-trimethyl ammonium
bromide (CTAB, a cationic surfactant), lecithin (a zwitterionic surfactant), and
GMS (a nonionic lipophilic surfactant). Although the addition of surfactant
was observed to result in a reduction in fat-droplet size, the degree of uniformity of the dispersion differed between emulsifiers. In contrast to previous reports that smaller more evenly dispersed fat droplets gave firmer cheeses
(Thomas et al., 1980; Shimp, 1985), Lee et al. (1995) found no relationship between processed cheese hardness and emulsion structure in the presence of
emulsifiers. They concluded that electrostatic interactions between the emulsifier and the protein played the major role in determining the rheological properties of the cheese. The anionic surfactant SDS gave the softest cheese, the
cationic surfactant CTAB the hardest. GMS and lecithin gave cheeses with
rheological properties little different from the control with no added emulsifier.
In the future, low molecular weight emulsifiers may play a greater role in controlling the texture of processed cheese.

7.8

Recombined, Concentrated, and Evaporated Milks

Recombined and concentrated milk products are produced for economic reasons. The cost effectiveness of transporting milk products that have been concentrated by removal of a proportion of the water phase, and the associated
changes in shelf life, make milk concentration a viable process. Similarly, it is
cost effective to transport dehydrated ingredients for recombination into milk.
The function of emulsifiers in these products is, primarily, to aid in the formation and stabilization of the emulsions. A secondary function, which is claimed
by many manufacturers of emulsifiers, is the effect that emulsifiers have on the
heat stability of milks and milk proteins.

200

Food Emulsifiers and Their Applications

7.8.1

Recombined Milk

Recombination of dairy ingredients into milk products is a popular and viable alternative to the export/import of fresh dairy products. It is particularly important
in countries where, for various reasons (e.g., transport delays, high temperatures),
the shelf life of fresh products prohibits their importation or local production.
In such cases, dried dairy ingredients are recombined close to the point of
sale, so as to reduce these problems.
Two approaches to recombining of whole milk can be used:

l. Recombination of anhydrous milk fat (AMF), skim milk powder (SMP),


and water
2. Reconstitution of whole milk powder (WMP) with water
In the past the latter process was, generally, less popular because of problems with the oxidative stability of the fat in the powder during storage.
Advances in gas packing of powders, more regular shipping, and use of cooler
storage facilities have removed this obstacle. Zadow (1982) noted that the
choice of whether to recombine or reconstitute WMP depends on the export
strategy of a particular manufacturer. During the late 1970s, an increase in the
production of reconstituted WMP was seen. This corresponded to a change
from a butter!SMP-orientated export industry to a cheese/WMP-orientated export strategy in countries such as New Zealand and Australia (Zadow, 1982).
In the recombination process, AMF, SMP, and water are recombined to give
a product with the same fat and protein content as whole milk.
The recombination process has been described by Kieseker (1983). The skim
milk powder is dissolved in the water at 40 to 55C. The fat is added in a molten
state, and the mixture is homogenized at 14.0 to 17.5 MPa for the first stage and
at 3.5 MPa at 55 to 60C in the second stage. The milk is then subjected to one
of three heat treatments: pasteurization at 72.2C for 15 seconds; UHT processing at 135 to 150C for 2 to 5 seconds; or in-can sterilization (e.g., 120C for lO
minutes). UHT processing can be by either direct steam injection for rapid heating or indirect heating in a plate or tubular heat exchanger.
Many manufacturers add low molecular weight emulsifiers to the formulation,
particularly mono- and diglycerides (Zadow, 1982; Kieseker, 1983; Sjollema,
1987). Emulsifiers in the form of phospholipids can also be added through the
practice of replacing up to 20% of the SMP with buttermilk powder (BMP)
(Zadow, 1982; Kieseker, 1983; Sjollema, 1987) to give an improved taste.
It is claimed that emulsifiers aid in the formation of the milk fat emulsion

Emulsifiers in Dairy Products and Dairy Substitutes

201

during homogenization. Recent research by Mayhill and Newstead (1992), however, suggests that little benefit in terms of emulsion formation and stability is
gained by their addition. In the case of mono-/diglyceride emulsifiers, it appears
that tradition dictates their presence in the formulation. It is possible that any reduction in creaming due to reduced fat-droplet size in the presence of emulsifier
is cancelled out by reduced emulsion stability caused by protein displacement.

7.8.2

Evaporated and Concentrated Milks

Evaporated and concentrated milks are produced by removal of water from natural or recombined milks. The technology used to make these products includes
evaporation under reduced pressure, reverse osmosis, ultrafiltration, and freeze
concentration (Knipschildt and Anderson, 1994; Varnan and Sutherland, 1994).
These concentrated milk products are more susceptible to heat coagulation,
when UHT processed or sterilized, than are normal concentration milks.

It has been known for some time that the heat stability of skim milk can be
altered by surfactant molecules (Singh and Creamer, 1992). Anionic surfactants such as SDS have been shown to shift the maximum in the heat stability/pH profile of skim milk to more acidic values and to give a marked increase
in maximum heat stability (Fox and Hearn, 1978). Cationic surfaetants such as
CTAB move the maximum heat stability to more alkaline values and give only
a slight improvement in heat stability at the maximum (Pearce, 1978; Shalabi
and Fox, 1982). The mechanism by which these changes occur is not known

for certain. It has been suggested that binding of the surfactant to casein micelles alters the surface charge, which leads to changes in heat stability (Fox
and Hearn, 1978; Pearce, 1978; Shalabi and Fox, 1982). This view is supported by the fact that nonionie surfaetants such as Triton X and Tween 80
have no effect on the heat stability of skim milk (Fox and Hearn, 1978).
Research into the effect of addition of SDS and CTAB on the heat stability of
milk proteins is useful only in helping to understand the process of heat coagulation. These surfactants cannot be added to milk products. In addition to
this, in most concentrated milk products and in whole milk, the fat globules
play a role in heat stability. Surfaetants would interact with both the fat-droplet
surface and the milk proteins. This makes the process of heat coagulation in
fat-containing milks more complicated than in skim milk.
The milk fat globule membrane is known to play a role in the heat stability of
milk. In nonhomogenized whole milk the fat globules have little effect on heat
stability (Singh and Creamer, 1992). However, after homogenization the heat eo-

202

Food Emulsifiers and Their Applications

agulation time decreases with increasing homogenization pressure (Singh and


Creamer, 1992). Obviously, this is an important observation since homogenization of milk is often essential so as to give adequate creaming stability.
It has been known for some time that lecithin can be used to increase the
heat stability of homogenized and concentrated milks (Maxcy and Sommer,
1954; Leviton and Pallansch, 1962; Hardy et al., 1985; Singh and Tokley,
1990; Singh et al., 1992). The mechanism oflecithin action has as yet not been
elucidated. Lecithin is known to displace protein from the fat-droplet surface
(Courthaudon et al., 1991; Dickinson et al., 1993a; Dickinson and lveson,
1993) and to complex with milk proteins (Barratt and Rayner, 1972; Korver
and Meder, 1974; Hanssens and van Cauwelaert, 1978). Hardy et al. (1985)
and McCrae and Muir (1992) believe that lecithin is incorporated into the adsorbed layer during and after homogenization. McCrae and Muir (1992) also
believe that lecithin/protein interactions play a role in heat stability of concentrated milks. Singh et al. (1992) have put forward the view that lecithin may
promote the formation of a complex between K-casein in the micelles and ~
lactoglobulin. The formation of the same complex can be promoted by preheating concentrated milks prior to the main heat treatment. This has been shown
by Newstead et al. (1977) to have a stabilizing effect on the heat stability of recombined evaporated milk. It is interesting to note that, despite evidence of
lecithin/protein interactions, Singh et al. (1992) have shown that the heat stability of skim milk is unaffected by lecithin addition. This is powerful evidence for the main stabilizing effect being fat-droplet based.

7.9

Other Dairy Applications of Emulsifiers

Emulsifiers have been added to other dairy products to exploit functional properties not normally associated with such emulsifiers. In recombined butter,
phospholipids are added as antispitting agents, to prevent fat spitting during
heating, and monoglycerides have been claimed to provide better "stand-up"
properties during storage (Kieseker, 1983).
Both sucrose esters and glycerol esters of fatty acids (monoglycerides) are
finding a wide range of novel uses. In addition to being good emulsifiers for use
in ice cream (Bucket al., 1986), they are known to improve the mouthfeel in
yoghurt (Farooq and Haque, 1992), inhibit microbial growth (Conley and
Kabara, 1973; Kato and Shibasaki, 1975; Shibasaki, 1979; Beuchat, 1980;
Kabara, 1983; Tsuchido et al., 1987), enhance the thermal death rate of bacte-

Emulsifiers in Dairy Products and Dairy Substitutes

203

ria and bacterial spores (Tsuchido et al., 1981; Tsuchido et al. 1983), and increase the heat stability of bovine serum albumin (Makino and Moriyama,
1991 ). It appears that these functions are a result of their ability to bind to proteins (Clark et al., 1992; Fontecha and Swaisgood, 1994).

7.10

Summary

Emulsifiers are very versatile food additives. They can be used as aids to
emulsion formation (e.g., in coffee whiteners/creamers and recombined products), or in contrast, as emulsion destabilizers as in ice cream, whipping
cream, and whipped toppings. These two functions rely on the classical ability
of emulsifiers to act as surface-active agents. In this way they can influence the
formation and stabilization of the fat-droplet adsorbed layer and the composition of this layer. This ability of emulsifiers to displace protein from the fatdroplet surface also, probably, accounts for the increase in heat stability of
concentrated milks when phospholipids are added.
In a similar vein, displacement of adsorbed caseinate by GMS in cream
liqueurs can be used to give increased acid stability to these products. A secondary function of the GMS in cream liqueurs is its ability to interact with proteins, thereby forming a weak gel in the aqueous phase. The associated increase
in viscosity gives improved creaming stability. The ability of the emulsifier SSL
to interact with the proteins in caseinate is also exploited in coffee whiteners.

The replacement of sodium caseinate in powdered coffee whitener is achieved


by using SSL. It has been hypothesized (Leo and Betscher, 1971) that this is possible because of the increased mechanical strength of a protein/SSL adsorbed
layer caused by emulsifier/protein interactions.
In processed cheese, the ability of charged emulsifiers to interact with proteins
in the cheese matrix may prove a useful way of controlling cheese texture. This
would introduce a way of reducing the concentration of emulsifying salts such as
mono- and polyphosphates. The final, but very important, function of some emulsifiers is their ability to act as initiators of fat crystallization. This is a particularly
important function in whipped products, and in combination with protein displacement forms the basis of the formation of the whipped foam structure.
A wide range of emulsifiers allowed for food use can be added to achieve the
above effects. Of late, consumer opinion has been focused on the "unnatural"
nature of synthetic emulsifiers. There is a slow push toward the replacement of
synthetic emulsifiers with natural emulsifiers such as milk and soy phospho-

204

Food Emulsifiers and Their Applications

lipids, and milk fat-derived mono- and diglycerides. The future may see a large
increase in the use of products such as BMP, which is rich in natural milk phospholipids as well as protein, and milk fat that has been enriched in mono- and
diglycerides by processes such as controlled glycerolysis of triglycerides.

Acknowledgments
I would like to thank the following people: Drs. Jeremy Hill, Peter Munro, and
Mike Boland (NZDRI) and Dr. Susan Euston (Massey University) for proofreading this manuscript; Dr. David Newstead (Milk Powder Technology
Section, NZDRI) for valuable discussion and comments on UHT fouling and
heat stability of concentrated milks; Dr. Siew-Kim Lee (Cheese Technology
Section, NZDRI) for comments on and suggestions for the section on processed
cheese; Mr. Bing Soo (Food Systems Section, NZDRI) for supplying information on coffee whitener/creamer; Mr. Tony McKenna (Food Science Section,
NZDRI) for supplying the electron micrographs used in Figures 7.1, 7.4, and
7.5; Vanessa Ellery (Graphics Unit, Information Centre, NZDRI) for preparation of Figures 7.2, 7.3, 7.6, and 7.7. Finally, I would like to thank the New
Zealand Dairy Board for permission to publish this work.

References
Abrahamsson, K., et al. (1988). Effects of homogenization and heating conditions on
physico-chemical properties of coffee cream, Milchwissenschaft, 43, 762-765.
Anderson, M., Brooker, B.E. (1988). Dairy foams, in Advances in Food Emulsions and
Foams (eds. E. Dickinson and G. Stainsby). Elsevier, London, pp. 221-255.
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Food Emulsifiers and Their Applications

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EIGHT

Applications of Emulsifers
in Baked Foods
Frank T. Orthoefer

8.1

Introduction

The development of emulsifiers or surfactants for bakery products has followed


the development of shortenings. The term "shortening" was initially used to refer to the fats used to "shorten" or tenderize baked foods. The composition of
shortening has progressed from natural fats to blends of oils, hydrogenated
fats, and hard fats. Shortenings may include additives such as emulsifiers, antioxidants, antifoams, and metal scavengers. In addition to their tenderizing
function, the shortening affects structure, stability, flavor, storage quality, eating characteristics, and eye appeal of baked foods. Many of the functional effects are due to or enhanced by the emulsifier added via the shortening. This
chapter will concentrate on the role of the emulsifier in baked foods.

8.2

History of Emulsified Shortenings

Historically, animal fats were used for bakery products because of their natural
plasticity and flavor (O'Brien, 1996). Lard was the preferred animal fat for its
pleasing flavor. Faced with excess supplies of cottonseeds and cottonseed oil,
vegetable shortenings were invented by the cottonseed industry early in this

211

212

Food Emulsifiers and Their Applications

century. Initially, cottonseed oil was blended with lard as a "lard compound"
or simply compound shortening. Hydrogenation as a process was invented in
1910. This new process allowed the development of substitutes for the semisolid (plastic) animal fats and permitted the creation of products with improved functional properties.
Along with this modification technique came improved methods for processing the oil, including refining, bleaching, and deodorization. The products
possessed improved oxidative stability, uniformity, and enhanced performance. With the development of lipid chemistry came methods for alcoholysis,
esterification, interesterification, and isomerization. These advances in lipid
chemistry brought new emulsifiers and shortening formulations. High-ratio
shortenings were introduced about 1933. These contained mono- and diglycerides that brought about finer dispersion of fat particles in the dough, giving
strengthened cake batters. These finer particle dispersions allowed higher
sugar levels, with increased water added, to produce sweeter tasting and more
tender cakes. High-ratio shortenings possessed excellent creaming properties.
Moist, higher volume, fine-grained, even-extured cakes were produced. Icings
were improved as well (Hartnett, 1977).
Emulsifier development also proceeded as well in the 1930s (Stauffer, 1996).
Specialty shortenings were the result. Commercial layer cakes, pound cakes,
cake mixes, creme fillings, icings, whipped toppings, bread, and sweet dough
shortenings were created. The development of specialty products resulted in improvements in processing and improved product performance for the retail, food
service, and food-processing industries. In addition to the traditional plastic
shortenings, liquid shortenings, fluid shortenings, and powdered products were
produced. All these shortenings involved formulation with emulsifiers.
The terms emulsifiers and emulsifying agents, surfactants and surface-active agents are used interchangeably in the literature. The terms "emulsifier"
and "emulsifying agents" are, strictly speaking, chemicals or compounds capable of promoting emulsification or stabilization of emulsions by their effect
on interfacial tension. Surfactants for foods may include not only emulsifiers
but also compounds with other functions such as protein or starch interaction.
In most cases, the terms are used interchangeably.
The role of the emulsifier and that of the shortening are intimately bound in
bakery products. Generally, the food emulsifiers for bakery products supplement and improve the functionality of a properly developed shortening.
Emulsifiers act as lubricants, emulsify fat in batters, build structure, aerate,

Applications of Emulsifiers in Baked Foods

213

improve eating quality, extend shelf life, modify crystallization, prevent sticking, and retain moisture. A list of emulsifiers commonly used in shortenings is
given in Table 8.1. The selection of or addition of an emulsifier to a shortening
base may significantly change the application of the shortening (Table 8.2).
Table 8.1 Emulsifiers used in shortenings
Mono- and diglyceride
Lecithin
Lactylaled monoglyceride
Calcium stearyllactylate
Sodium stearoyllactylale
Propylene glycol monoesters
Diacetyl tartaric monoglycerides
Ethoxylaled monoglycerides

Sorbilan monostearate
Polysorbate 60
Polyglycerol esters
Succinylated monoglycerides
Sodium stearoyl fumarale
Sucrose esters
Stearoyllactylate

Table 8.2 Examples of nonemulsulfied and emulsified shortenings


Nonemulsified

Emulsified

All-purpose
Puff pastry
Pie crust
Cookie
Danish roll-in
Donut fry

Cake and icing


Household
Filling
Cake mix
Yeast raised
Specialty cake

8.3

Emulsifier Function in Baked Goods

Baked goods without emulsifiers have been described as tough, dry, stale,
leathery, or tasteless (Brandt, 1996). Current processing, distribution, and
storage of baked goods requires the use of additives that maintain quality and
freshness. Emulsifiers are commonly used in many food products. These supplementary materials or food additives are used to

Compensate for variations in raw materials

Guarantee constant quality

Produce attractive products

Preserve freshness and eating properties

Facilitate processing (Schuster and Adams, 1984)

214

Food Emulsifiers and Their Applications

Emulsifiers in nearly all applications refers to their ability to emulsify oil in


water. This is also true in some bakery applications. However, emulsification is
often of secondary importance. Activity as surface-active agents is most desirable. Starch complexing, protein strengthening, and aeration are the primary
functions. Fat sparing effects are also of importance. The average composition
of wheat flour is

Fraction

Percent

Starch
Protein
Pentosan
Lipid
Crude fiber
Ash

70.0-75.0
11.5-12.5
2.0-2.5
l.0-1.5
0.2
05

The interactions between proteins, carbohydrates, and lipids are significant


for the processing of wheat flours. The interactions between emulsifiers and
flour components are multifaceted and account for the improved functionality
and performance of baked products.
Surfactants used in bakery products are regulated in most countries. Each
country may differ in their regulations but the usage level and applications are
well defined. European Economic Community (EEC) number and U.S. FDA
Code of Federal Regulation (21CFR) for the most common food emulsifiers are
shown in Table 8.3. The specification and assay procedures of all emulsifiers
presented in this chapter are published in the Food Chemicals Codex
(Organization of the Food Chemicals Codex 1981).
Baked products are the largest users of food emulsifiers. Yeast-raised and
chemically leavened products are the most important segments. Other uses
include cookies, crackers, pasta, and snacks. Recent figures indicate that
about 400 million pounds of emulsifiers are used in the food industry. The
bakery industry accounts for 50% of the total food emulsifier market
(Brandt, 1996). Annual growth in the production of food emulsifiers is estimated at about 3%.

Applications of Emulsifiers in Baked Foods

Table 8.3

Regulatory status of emulsifiers. (From O'Brien, 1996, p. 517.)

Emulsifier
Monoglycerides and diglycerides (GRAS)
Succinyl monoglyceride
Lactylated monoglyceride
Acetylated monoglyceride
Monoglyceride citrate
Monoglyceride phosphate (GRAS)
Stearyl monoglyceride citrate
Diacetyl-tartrate ester of monoglyceride (GRAS)
Polyoxyethylene monoglyceride
Propylene glycol monoester
Lactylated propylene glycol monoester
Sorbitan monostearate
Polysorbate 60
Polysorbate 65
Polysorbate 80
Calcium stearoyllactylate
Sodium stearoyllactylate
Stearoyllactylic acid
Stearyl tartrate
Sodium stearyl fumarate
Sodium Iaury! sulfate
Dioctyl sodium sulfosuccinate
Polyglycerol esters
Sucrose esters
Sucrose glycerides
Lecithin (GRAS)
Hydroxylated lecithin
Triethyl citrate (GRAS)

8.4

215

U.S. FDA
(2ICFR)

EEC
nlllllber

182.4505
172.830
172.852
172.828
172.832
182.4521
172.755
182.4101
172.834
17.854
172.850
172.842
172.836
172.838
172.840
172.844
172.846
172.848

E47l
E472
E 472
E472
E472
E472
E477
E491
E435
E 436
E433
E 482
E 481
E483

172.826
172.822
172.810

172.854
172.859
184.1400
172.814
182.1911

E 475
E 173
E474
E322
E322

Role of the Shortening

A shortening when mixed into a dough interupts the development of the gluten
structure. Literally, the structure is "shortened" and the baked product is tender. Overall, the shortening also contributes to the quality of the finished product by imparting a creamy texture and rich flavor, tenderness, and uniform
aeration for moisture retention and size expansion. Fat-based products are formulated and processed for plasticity to allow spreadability and to disperse

216

Food Emulsifiers and Their Applications

thoroughly and uniformly in a dough, icing, or batter over a wide temperature


range. The ability of the fat to disperse in streaks and films helps to lubricate
the structure of the dough during mixing and thereby prevents the starch and
protein components from compacting into a dough mass.
The characteristics of the fat that are important for shortening formulations
include melting point, stability, solid fat index, and plasticity. Plasticity is used
to define the characteristics of shortening that are functionally most important.
Shortenings are processed to various plasticity ranges (Weiss, 1981).
Narrow-plastic-range products have a steep solids profile and melt rapidly.
These are commonly used in cream icing products or as filler fats for hard
cookies where melting near body temperature is required. Wide-plastic-range
shortenings contain 15 to 30% solids over a broad temperature range, and resist breakdown during creaming. Their plastic nature enables them to spread
readily and combine thoroughly with the other solids or liquids without cracking, breaking, or having liquid oil separating from the crystalline fat.
Commercial products are prepared by carefully cooling, plasticizing, and tempering correctly formulated blends of melted fats and oils.
Crystal size has a major influence on the rheological properties of plastic
shortenings. A small crystal size with a large surface area is required to effectively bind the liquid oil present in a shortening formulation. Typical crystal
sizes are from 5 to 9Jlm (P. Chawla and J.M. deMan, 1990).
Crystal size is controlled by source of the oil (O'Brien, 1996). The smaller crystalline form is referred to as Wand the larger form is ~- Plastic shortenings in the
Wconfiguration consist of small, uniform, needle-like crystals with a smooth texture and the ability to aerate well, and they have excellent creaming properties.

8.5

Role of the Emulsifier

The emulsification of the shortening in bakery products is promoted with the


addition of surface-active agents (surfactants, emulsifiers). Much of the work
on shortenings has centered on the addition of an emulsifier or emulsifier system to an all-purpose shortening base, although specialty liquid, narrow-plastic-range, and special-purpose emulsified products have been produced.
Emulsifiers are used extensively in bakery products, especially breads and
cakes. Some of the benefits are

l. Increased shelf life


2. Imparted tenderness and improved flavor release

Applications of Emulsifiers in Baked Foods

217

3. Reduced mixing time and improved mixing tolerance


4. Improved machinability
5. Improved gas retention
6. Improved water absorption
7. Improved volume
8. Improved hydration rate of flour and other ingredients
9. Better texture and symmetry
10. Reduced egg and shortening usage

8.5.1

Monoglycerides and Derivatives

Monoglycerides are prepared by reacting glycerine with edible fats and oils or
fatty acids in the presence of a catalyst (Henry, 1995). The important characteristics are melting point and monoglyceride content. The monoglyceride content varies from 40 to 95%. Two crystalline forms are generally present: alpha
and beta. The alpha form is the most functional in bakery products. The major
variables involved in the production of monoglycerides are source of the fat,
monoglyceride content desired, iodine value or degree of unsaturation, and
fatty acid composition. Approximately 40 million pounds (18 million kg) of
monoglycerides are used in the United States in yeast-raised bakery products
(WH. Knightly, 1988). An equal amount was believed to be used in cakes, icings, and other applications. In addition to the improved aeration and sugarholding capacity of cakes formulated with monoglycerides, breads were found
to have an improved shelf life due to retarded staling rate. Various techniques
were applied to improve monoglyceride formulations including other emulsifier types. Today, plastic, hydrated, powdered, and distilled monoglycerides
are used in bakery applications.
In addition to their antistaling effects, monoglycerides in bakery products
result in

Reduction of interfacial tension

Improved dispersion of ingredients

Increased aeration

Greater foam stability

Modification of fat crystals

Several derivatives of monoglycerides are prepared for bakery applications


(Figure 8.1). These are of two general types. One is considered as dough

218

Food Emulsifiers and Their Applications

When R equals

Emnlsifier

HO-

Glycerol monosterate (GMS)

Succinyl monoglyceride (SMG)

CH CH-C00-

31

Lactylated monoglyceride (LacGM)

OH

CH 3

-coo-

Acetylated monoglyceride (AcMG)

OOCCH

Diacetyl-tartaric acid ester of

-QOC-cH-CH-COQ-

monoglyceride (DATEM)

OOCH 3

Polyoxyethylene monoglyceride or
ethyoxylated monoglyceride
(EMG)

H-

Propylene glycol monoester


(PGME)

Figure 8.1

Monoglycerides and derivatives.

Applications of Emulsifiers in Baked Foods

219

strengtheners and includes SMG (succinylated monoglycerides), EMG


(ethoxylated monoglycerides), and DATEM (diacetyl tartaric acid esters of
monoglycerides). They also are used as emulsifiers, starch- and protein-complexing agents, and foam stabilizers. The other is the alpha-tending emulsifiers
and includes GMS (glycerol monosterate), LacGM (lactylated monoglycerides),
AcMG (acetylated monoglycerides), and PGME (propylene glycol monoesters).
The alpha-tending emulsifiers, normally used in cake production, contribute to
the emulsification of the shortening in the water phase as well as incorporating
air into the fat phase. These are believed to form a solid film at the oil/water interface resulting in a stable emulsion and preventing the liquid oil from interfering with aeration during cake batter mixing.

8.5.2.

Sorbitan Emulsifiers

Sorbitan monostearate is an oil-soluble, low-HLB nomomc emulsifier.


Reaction of the sorbitan esters with ethylene oxide results in the formation of
the polyoxyethylene sorbitan monostearate or polysorbate emulsifiers (Figure
8.2). Sorbitan esters are excellent emulsifiers for icings, improving aeration,
gloss, and stability. They function as emulsifiers, aerating agents, and lubricants in cakes, toppings, cookies, and crackers. The polysorbate 60 emulsifier
is used as a dough strengthener at about 0.2% of flour weight. Polysorbate 60
is also used in combination with glycerol monostearate and propylene glycol
monostearate in fluid cake shortenings.

8.5.3

Anionic Emusifier

The anionic emulsifiers include SMG, DATEM (see (Figure 8.1), and other lactic acid derivatives (Figure 8.3). Sodium stearoyl lactylate (SSL) is the most
widely used in the United States. The calcium version, insoluble in water, is
also widely used. Both are dough strengtheners.
SSL may be added as a stabilizer to hydrated monoglyceride preparations.
Other benefits from the lactic acid emulsifiers are antistaling, aeration, and
starch and protein complexing.

8.5.4

Polyhydric Emulsifiers

The polyhydric emulsifiers include the polyglycerol esters and sucrose esters
(Figure 8.4). Both have multiple applications as emulsifiers for foods in general and in bakery products, particularly the sucrose esters. The sucrose esters
are common food emulsifiers in Japan. They provide emulsifying, stabilizing,

220

Food Emulsifiers and Their Applications

Where
Sorbitan monostearate
R1 = H (CH 2 CH 2 0)n-

Polyoxyethylene (20) sorbitan monostearate

R2 = H

(Polysorbate 60)

R3 = H

R1 = H(CH2 CH 2 0)nR2 and R3 = H3C (CH 2) 16CO-

Figure 8.2.

Polyoxyethylene (20) sorbitan tristearate


(Polysorbate 65)

Sorbitan esters and derivatives.

and conditioning effects in baked goods. Eight hydroxyl groups that may be esterified are present in sucrose. As expected, the degree of esterification affects
the hydrophylic lipophilic balance (HLB) of the sucrose ester (Table 8.4). The
product is considered a noncaloric fat substitute when six or more of the hydroxyls are sterified (Olestra by Proctor & Gamble).

8.5.5

Lecithin

Lecithin is a coproduct of soybean oil and, to a limited extent, of other vegetable oil refining (com oil and rapeseed oil). It is obtained by water-washing
crude vegetable oil and separating and drying the hydrated gums. The crude
material is very dark colored and viscous, consisting of a variety of phospholipids (Figure 8.5). In addition to the phospholipids, these are also present

Applications of Emulsifiers in Baked Foods

221

CH 3

Sodium stearyllactylate (SSL)


{also in calcium form)

II

II

H3C (CH 2)17-0-c-CH-CH-C-o8NJ)

Sodium stearyl fumarate

Sodium lauryl sulfate (SDS)


Figure 8.3. Anionic surfactants.

triglycerides, tocopherols, and glycolipids. Various purified grades are produced by bleaching and fractionating as well as by chemical modification
(Schmidt and Orthoefer, 1985). Commercial lecithin products are specified
based on their acetone-insoluble fraction (a measure of the phospholipid content), viscosity, and color. Lecithin is present in other sources such as egg yolk,
butter, beans, nuts, etc. Lecithin is usually the least expensive emulsifier.

8.6

Emulsifier Interaction with Bakery Components

Emulsification and lubrication by the emulsifiers accounts only partially for


the beneficial effects observed when they are added to baked products.
Proteins and lipids also contribute to the functional properties of the flour.
Emulsifiers have been shown to interact with the various components present
as well as with added ingredients.

222

Food Emulsifiers and Their Applications

OH

CH 2-CH-cH 3

[0-CH 2-CH-CH 2-OJ n


1

OH

0-H C-HC-CH
2

OH

Polyglycerol monostearate

II

CH 2-0-C-(CH 3 ) 16CH 3

0
H

HO
H

OH

OH

Sucrose diester

Figure 8.4

8.6.1

Starch

Starch exists in a helical, coiled configuration with about six residues per tum.
This structure is basically a hollow cyclinder that has a hydrophyllic outer surface and a hydrophobic inner core. The inner space is about 45 nm in diameter. Straight-chain, alkyl molecules such as stearic acid will fit in the inner

Applications of Emulsifiers in Baked Foods

Table 8.4

223

Sucrose ester surfactants. (From Stauffer, 1996, p. 576.)

Percent
monoester

Percent
diester

71
61
50
46
42
33

24
30
36
39
42
49

Percent
triester

Percent
tetraester

0
1
2
2
2
2

12
13
14
16

HLB

15
13
ll

9.5
8

CH 2-0-R 1

r~~
CH -0-P-Q-R
2

lb

(R 1 and R2 are fatty acids)


If R2

Then

= Choline ethanolamine inositol serine


Phosphatidylcholine
Phosphatidylethanolamine
Phosphatidyl inositol
Phosphatidylserine

Figure 8.5.

Lecithin.

space. The n-alkyl portion of emulsifiers such as present in GMS form a complex with the helical regions of the starch. It is this complex that is thought to
retard starch crystallization in bread, often called "retrogradation," slowing
the staling process.
The influence of emulsifiers on starch consists of (i) affect on the rate of
gelatinization, the gelatinization temperature, the peak viscosity, and the gel

224

Food Emulsifiers and Their Applications

strength and (ii) complex formation by the starch molecules. In studies of


starch pastes with monoglycerides, the maximum complexation occurs with
monopalmitin (Lagendijk and Pennings, 1970). Longer and shorter saturated
fatty acid monoglycerides reacted to a lesser extent. Unsaturated fatty acid
monoglycerides react to a lesser extent because of the bend in the fatty acid
chain due to the unsaturated linkage (Hahn and Hood, 1987).
Surfactants modify the gelatinization behavior of starch. DATEM is generally found to be less interactive than GMS or SSL. GMS raises the swelling
temperature and results in increased paste viscosity. SSL also increases paste
viscosity (Schuster and Adams, 1984). Overall the interaction between emulsifier and starch takes place at the surface of the granule, and the starch/surfactant complex apparently stabilizes the granule, retarding water penetration and
swelling as the temperature is increased.
During breadmaking, only small amounts of emulsifiers are bound to starch
in the sponge stage and during mixing. Binding does not occur until the temperature is increased to near the gelatinization temperature. The formation of
starch complexes is ascribed principally to the amylose portion of the starch.
Both the degree of interaction and solubilities of the complexes are dependent
upon the emulsifier type.

8.6.2

Protein

The wheat flour proteins, gliadin and glutenin, form a viscous, colloidal complex known as "gluten" when mixed into a dough. Lipids are also involved in
the formation of the gluten complex. The properties of the gluten are influenced by the lipids and the emulsifiers present. Lipophilic parts of surfactants
interact with hydrophobic regions of proteins, contributing to unfolding or denaturation of the protein. Generally, surfactants contribute to protein denaturation, enhancing interfacial absorption and emulsion stabilization.
Most dough strengtheners are anionic surfactants. The association of the
lipophilic portion to the hydrophobic protein portion incorporates the negative
charge into the complex, promoting aggregation in the dough. The overall effect is aggregation of the gluten protein and an increase in dough strength.
The ionic surfactants induce protein insolubilization, resulting in increased
viscosity and elasticity of the dough. Nonionic surfactants disrupt the hydrophobic portion of the protein, leading to reduced dough viscosity and elasticity and increased protein extractability. A blend of emulsifiers generally
provides the best dispersibility and functionality.

Applications of Emulsifiers in Baked Foods

8.6.3

225

Lipids

Wheat flour contains 1.4 to 2.0% lipids divided into free (0.8 to 1.0%) and
bound (0.6 to 1.0%) forms. They may be further divided into nonpolar (50.9%)
and polar (49.1 %) forms. The bound lipids exist as starch inclusion complexes. The nonstarch lipids, about 85% of the total, participate in the chemical, physical, and biochemical processes important for the preparation of
baked goods. The nonstarch lipids consist of glycolipids, phospholipids, and
stearyl esters. Interaction between nonstarch lipids and emulsifiers is limited.
Likely, added lipids merely influence or substitute for each other.
Non-polar-lipid addition to untreated flour gives a deterioration of baking
properties (Schuster and Adams, 1984). Addition of polar lipids to untreated
flour increases loaf volume in breadmaking. The improvement is likely based
on the effects of galactolipids and phospholipids. Emulsifiers may interact with
the water phase of the dough, forming associated lipid-water structures with
free polar flour lipids (Krog, 1981). Emulsifiers may compete with naturally
occurring lipids in wheat flour for the reactive groups of the wheat flour dough.
Their effect on protein components was modulated as a consequence.

8.7

Applications in Baked Goods

8.7.1 Yeast-Raised Products


The functions of emulsifiers in yeast-raised products are dough conditioning
and strengthening as well as crumb-softening. Emulsifiers are also used.

8.7.1.1

Dough Conditioning. There is no precise definition of dough condi-

tioners. The term is commonly used to describe adjuncts that aid in the development of less tacky, more extensible doughs that are processed through
machinery without tearing or sticking or that result in a product of finer crumb
structure and improved volume and symmetry. These may be summarized as

Increased mixing and machining tolerance of the dough

Increased tolerance to ingredient variations


Diminished knockdown during handling

Assists maximum dough absorption


Reduced shortening requirements
Improved loaf volume, structure, texture, and other quality characteristics

226

Food Emulsifiers and Their Applications

Extended keeping quality


Facilitates variety bread production

In the production of yeast-raised products, the mixing of the dough results


in gluten/gluten bonding through disulfide linkages. Development of the linkages is often incomplete, resulting in a weak dough structure. Gas that is produced by the yeast escapes through the weak portion of the gluten films. Cells
having weak gluten walls have a tendency to collapse.
Dough-strengthening emulsifiers increase the degree of gluten/gluten binding
sites and/or bridges that supplement disulfide linkages. This results in stronger,
developed gluten films. As a result the benefits from dough conditioners are

Improved tolerance to variation in flour quality

Drier doughs with greater resistance to abuse

Improved gas retention giving lower yeast requirements, shorter proof times,
and greater finished product volumes

Uniform internal grain, stronger side walls, and reduction of "cripples"


Reduced shortening requirements without loss of volume, tenderness, or
slicing ease

The dough conditioners recognized as highly functional dough strengtheners are calcium stearoyl lactylate, ethoxylated monoglycerides (EOM), polyoxyethylene sorbitan monostearate (PS 60), succinylated monoglycerides
(SMG), and sodium stearoyl lactylate (SSL) (Tenny,l978). Comparative loaf
volumes found for the various conditioners are shown in Figure 8.6, for fully
proofed dough shocked to mimic abuse in production.
Dough conditioners are available in plastic, powdered, and liquid forms
that may he added to the sponge, brew, or dough. The use levels are normally
in the range of 0.25 to 0.5% of flour weight. Often the dough conditioners are
combined with crumb softeners. For example, EOM, PS 60, and SMG are
blended with monoglycerides, a crumb softener.

8.7.1.2

Crumb-Softening.

Emulsifiers that complex with starch are referred


to as "crumb softeners." Although there is a controversy over the mechanism of
activity, evidence exists that a primary effect is to complex with amylose, the linear starch fraction. The staling phenomena is generally believed to result from
amylose crystallization. Amylose leaches from the starch granule during dough
preparation and baking. The amylose polymers associate upon cooling, forming a
rigid gel within lO to 12 hours. Amylopectin, the branched chain starch fraction,

Applications of Emulsifiers in Baked Foods

227

2900

2800

1-

2700

2600

_fl
2500 -

1-

r-----

f-----

- :--

f-

'.:

- :'I!

r--: :-

f-

1--

f-

~l ~
r---:

1---

00

NO

ADD

.10 .25 .50

CSL

.10 .25 .50

EOM

.10 .25 .50

PS-60

.10 .25 .50

.10 .25 .50

SMG

SSL

%Flour, wt

Figure 8.6 Comparative loaf-volume response produced on abused dough by CSL,


EOM, PS-60, SMG, and SSL. (From Tenney, 1978.)

crystallizes more slowly, resulting in firming of the bread in 3 to 6 days. Crumb


softeners tend to result in less free amylose from the baking process; therefore,
less free amylose is avaliable to form a rigid gel. The emulsifier "softens" the initial crumb. No change occurs with the amylopectin fraction with softener addition. It gradually crystallizes to form a firmer texture. Treated bread stales at the
same rate as untreated bread in the case of amylopectin.
Comparison of softeners as a function of compressibility after 96 hours of
storage is shown in Figure 8. 7. The most effective softeners are the lactylates
and SMG. Plastic mono- and diglycerides and hydrated distilled monoglycerides are also effective. The polysorbate, EOM, and lecithin had little starch
complexing ability. There is an apparent dual functionality of conditioner and
softener for the lactylates and SMG.
Use levels of softeners vary with the market. The most commonly used crumb
softeners are the water emulsions, or hydrates, of mono-/diglycerides. The hydrates contain 22 to 25% solids and are used from about 0.5 to l% of flour weight.
The hydrates are significantly more functional. Water-dispersible distilled monoglycerides are also sold. These blends use sufficient unsaturated monoglycerides
to promote rapid hydration in the water of the sponge, brew, or dough.

228

Food Emulsifiers and Their Applications

Crumb compressibility (0.1 mm)


180

170

160

150

140

o_

12

11

130

NO
ADD

r---. ; r---

~- ~
.10 .25 .50
CSL

r---

10.25.50
EOM

~!I_

Jl

.10 .25 .50


PS-60

_Jl ~Jr---

.10 .25 .50


SSL

r-~ r-J

JJ

10.25 .50

.10 .25 .50

SMG

Mo-Di

.10 .25 .50


LSC

.10 .25 .50 1.0


Dis. M . H.

%Flour, wt
Figure 8.7

Relative crumb-softening effect in bread by CSL, EOM, PS-60, SSL, SMG,


Mo-Di (54% mono- and diglyceride), LEC (lecithin), and Dis. M.H. (22% solids distilled monoglyceride hydrate). (From Tenney, 1978.)

8.7.1.3

Emulsifier Blends.

Lecithin has been used in breads and baked

goods longer than any other emulsifier. It was shown to give higher ductility
through interaction with the gluten. Other activity from the lecithin is delayed
staling and shortening reduction. A synergistic effect also occurs with lecithin
and monoglycerides. The monoglycerides produce better crumb grain, softer
bread, and higher loaf volumes. Ethoxylated monoglycerides with monoglyceride is an effective dough conditioner. The negative effects of liquid oils in
place of shortening in bread preparation is overcome with the combination.
DATEM also acts as a dough conditioner, shortening saver, and antistaling
agent in combination with glycerol monostearate. Others include SMG, sucrose esters, polysorbate 60, SSL, and CSL. The SSL and CSL can form complexes with gluten acting as a dough strengthener.
The direct and indirect action of the emulsifier begins with dough preparation and ends with oven baking and storage (Figure 8.8). The first stage begins

Applications of Emulsifiers in Baked Foods

Improvement of
wettability
Stabilization of
distributed phases

Mixing

Fermentation
Scaling, kneading,
moulding

Baking

Storage

229

Decrease of mixing time and


mixing speed
Reducing shortening levels
Improvement of mixing tolerance
Improvement of machinability

Improvement of gas-retaining
properties
Shorter fermentation
Greater shock-tolerance

Improvement of gas-retaining
properties
Improved loaf volume
Better texture
Better crumb grain
Better uniformity
Decrease of water loss

Improvement for crumbsoftness


Longer shelf life

Figure 8.7 Influence of emulsifiers on production and quality of baked products. (From
Schuster arul Adams, 1984.)

with wetting, and dispersing activity then follows with interactions with flour
components during mixing and in the baking process itself.

8.7.2
8.7.2.1

Chemically Leavened Products

Cakes. The role of emulsifiers in layer cakes or snacks cakes is diverse. They include aeration, emulsification, and crumb-softening.
The aerated structure of batters depend on whipped-in-air and carbon diox-

230

Food Emulsifiers and Their Applications

ide from the leavening agent. The use of emulsifiers lowers the surface tension
of the aqueous phase, improving the amount of air that can be whipped into the
batter. Large amounts of finely divided air cells are important for development
of uniform grain (Randleman et al.,1961). The dissolved C0 2 evolves at air
cell sites and does not spontaneously form bubbles. If the original batter contains many small air cells, the final cake will have a larger volume and fine
(close) grain. The creaming of the sugar and shortening have a major influence
on air incorporation. The incorporation of monoglycerides in the plastic shortening (2 to 5% alpha-monoglycerides) ensures numerous small air cells being
created during beating or creaming.
Cake batter is, of course, an aerated emulsion. The integrity of the air cells
determines cake volume and uniformity. Shortening is an antifoam that disrupts
foam cells. Emulsifiers, however, coat the exterior of the fat particles protecting
the integrity of the air cell (Wooten, 1967). Proper emulsifier selection has permitted the use of liquid oil where only shortening could previously be used.
Light, tender, moist cakes are preferred by the consumer. Emulsifiers provide the desired aeration, emulsification, and crumb-softening. Crumb-softening in cakes is a function of moisture retention, shortening activity, and
starch-complexing. The starch-complexing is the same as for breads. The
emulsifiers complex with the starch to soften the product.
Several types of emulsifiers are used in cakes. Propylene glycol monoester
(PGME) is utilized at 10 to 15% of the shortening. Monoglycerides and mixtures of lactated monoglycerides with PGME are also used in cake mixes.
In bakers' cakes, a large number of emulsifiers are used, depending on the formula, production equipment, and labeling requirements. With soybean oil as the
shortening, a hydrated blend of emulsifiers such as PS 60, SSL, sorbitan monostearate, and distilled monoglycerides work well. Fluid shortenings are produced
containing lactated monoglycerides. The traditional bakers'-cake system is a plastic shortening with 5 to 10% monoglycerides (3% alpha-monoglyceride content).
Packaged cake mixes often use emulsified PGME at 10 to 15% of the oil. The
cakes are unusually tender and are not suitable for commercial cake production.
Emulsified cake shortenings are also used for cake donuts. The degree of air
entrapment during the creaming phase determines the grain in the final donut.
8.7.2.2

Cookies and Crackers. Although emulsifiers are used to only a limited

extent in cookies and crackers, they play a significant role in (i) controlling
spread, (ii) improving cutting and appearance, and (iii) improving texture. The

Applications of Emulsifiers in Baked Foods

231

limited use probably stems from the reluctance to change the successful formulas in this segment of the baking industry. New products utilizing emulsifiers are receiving greater acceptance since they have been shown to play
functional roles.
Certain emulsifiers have the ability to change the degree of spreading of
cookie dough as it is baked (Table 8.5). The effect likely occurs because of altering of the viscosity of the dough. Cookie dough with SSL shows increased
spread compared to a nonemulsified control (Rusch, 1981). The SSL may interact with the starch granule, delaying hydration of the granule and subsequent gelatinization (Tsen et al., 1973).
Lecithin may be used to produce a drier dough that machines better and
that improves release from the rotary die surface. Use is from 0.25 to 0.7% of
the flour. Part of the effect may simply be reduction of available water because
of lecithin hydration.
Lecithin that is highly fluidized with other oils or fatty acids is widely used
as release agents in cookie baking due to improved release from rotary dies.
Heat-resistant lecithins such as those modified with acetic anhydride are especially adaptable to the application. Lecithin is used in cookie and cracker formulations at 0.25 to 1.0% of flour weight. It may be added with the shortening
at the creaming stage or simply combined with the shortening when rotated.
Antistaling is of less significance in cookies and crackers since they are of

Table 8.5 Spread ratios of cookie


doughs with different emulsifiers. (From
Tsen et al., l973.)
0.5% additive
Monoglyceride
Ethoxylated monoglyceride
Sodium stearoyl fumararale
Sodium stearoyllactylate
Sucrose monopalmitate
Sucrose mono- and distearate
Sucrose distearate
Sorbitan monostearate
Polyoxylated (20) sorbitan
monostearate
Succinylated monoglycerides

Spread
ratio
8.3
8.8
10.4
10.0
9.8
9.6
9.7
9.2
9.3
9.2

232

Food Emulsifiers and Their Applications

lower moisture content. The greasiness of high shortening levels is reduced by


the addition of small amounts of lecithin. Lecithin in general gives a "drier"
dough with equivalent moisture and shortening levels. The drier dough is more
machinable. Other benefits attributed to lecithin are reduced mixing times and
dough development with more tender cookies.
SSL is promoted to improve the efficiency of shortening in cookie or cracker
formulations. SSL when incorporated into dough at 0.25% of the flour produces a finer grained, more uniform pattern of surface cracks. The resistance
to shear (firmness) decreases, with improved eating quality, and permits shortening reduction (Tenney, 1978). Levels are 0.25% SSL in cookies and 0.1% in
crackers based on flour weight.

8.7.3

Extruded Snacks/Cereals

Extrusion-cooked snacks, pastas, and cereals often include emulsifiers in the


formula. Gelatinization of the starch occurs during the cooking/extrusion step.
Monoglycerides and SSL have been found to reduce the energy required for
the extrusion and to obtain a desirable texture in the final product. When
monoglycerides are added, they influence the appearance and properties of the
extrudate such as smoothness of the extrudate surface and fine pore structure.
Use levels are typically 0.25 to 0.5% of the starch weight and is added at the
dough makeup stage.

8.7.4

Cream Icings

Cream icings are prepared by creaming sugar with fat, then adding flavor, egg
white, and perhaps a small amount of water. The emulsified shortening generally contains 2 to 3% alpha-monoglyceride. Polyoxyethylene sorbitan monostearate at 0.5% is included in some icings to assist in air incorporation.
Propylene glycol esters when incorporated into the shortening produce icings
with excellent gloss and gloss retention.

8.7.5

Fat-Free Bakery Products

The fat-free or low-fat foods are marketed in almost every segment of the food
industry. In most instances, there is no one solution for removal of fats from the
formulation. Skillful formulation using fat replacers, emulsifiers, bulking
agents, flavors, and other ingredients has been applied to fat replacement.
Low-fat and fat-free cakes have been produced using additional emulsifiers
in conjunction with starch-based fat replacers and gums or hydrocolloids for

Applications of Emulsifiers in Baked Foods

233

moisture retention and functionality. PGME and DATEM products have assisted in formula development.
Emulsifiers are not generally regarded as fat substitutes or replacers.
Emulsifiers affect the texture and mouthfeel by their effect on surface interactions. The caloric value of emulsifiers varies depending on their actual composition and digestibility. They tend to have fat-like properties through their
hydration, water-binding, and dispersing effects in processed foods. The general functions of emulsifiers in low-fat or no-fat applications are to

Prevent separation of components

Reduce size of fat globules and improve dispersion of remaining fat

Provide a fat-sparing action

Provide texture perception of higher fat levels

Texturize, provide lubricity

Complex starches and proteins

Mono- and diglycerides are the most used emulsifiers. Distilled monoglycerides have a caloric advantage over the lower mono-containing preparations.
Other emulsifiers in reduced fat products include the polysorbates, polyglycerol esters, and sorbitan monoesters.
Many noncaloric fats have been synthesized. Sucrose polyesters (OlestraTM
from Proctor & Gamble) is probably the best known. Emulsifier use in products utilizing the noncaloric, synthesized fat replacers is very likely similar to
that utilizing the traditional caloric version.

8.7.6

Release Agents

A separate application of emulsifiers in bakery products, although not incorporated into the dough, is release agents or pan sprays. Lecithin is the primary
emulsifier used. Often the pan sprays are formulated with an oil in combination with mold inhibitors and lecithin. One to 6% lecithin is added. Modified
lecithins that exhibit improved heat stability may used. The pan spray may
simply be brushed or sprayed to achieve a thin film promoting easy release of
baked products from pans or belts.

8.8

Summary

The market for emulsifiers in bakery products continues to grow. Reduced-fat


products have provided the more recent surge in utilization. Producers of

234

Food Emulsifiers and Their Applications

emulsifiers as well as bakeries themselves, as with many of our industries,


have undergone consolidation. Fewer producers have placed new requirements on the final product that is transferred to the ingredient supplier, in this
case the supplier of the emulsifier. The function of the emulsifier is even of
greater importance. Growth in food service furthers the need for bakery products having desirable sensory characteristics and meeting the demands of today's marketplace.

References
Brandt, L. (1996). Emulsifiers in baked goods, Food Product Design, Feb., pp. 64-76.
Chawla, P., deMan, J.M. (1990). ]. Am. Oil Chem. Soc., 67, 329.
Hahn, D.E., Hood, L.F. (1987). Factors influencing com starch lipid complexing, Ger.
Chem., 64,81-85.
Handleman, A.R., et al. (1961). Bubble mechanisms in thick foams and their effect on
cake quality, Ger. Chem., 38, 294.
Hartnett, D.I. (1977). ]. Am. Oil Chem. Soc., 54, 557.
Henry, C. (1995). Monoglycerides: the universal emulsifier, Ger. Foods World, 40(10),
734-738.
Knightly, W:H. (1988). Ger. Foods World, 33,405-412.
Krog, N. (1981). Theoretical aspects of surfactants in relation to their use in breadmaking, Ger. Chem., 58, 158--164.
Lagendijk, J., Pennings, H.J. (1970), Ger. Sci. Today, 15, 354-356, 365.
O'Brien, R.D. (1996). Shortening: types and formulations, in Bailey's Industrial Oil and
Fat Products (ed. Y.H. Hui), 5th ed., Vol. 3, pp. 161-193, Wiley, New York.
Rusch, D.T. (1981). Emulsifiers: uses in cereal and bakery foods, Ger. Foods World,
26(3), 1ll-ll5.
Schmidt, J.C., Orthoefer, F. (1985). Modified lecithins, in Lecithins (eds. B.F. Szuhaj
and G.List), Chap.lO, pp. 203-213, Am. Oil Chern. Soc., Champaign, IL.
Schuster, G., Adams, W.F. (1984). Emulsifiers as additives in bread and fine baked
products, in Advances in Cereal Science and Technology (ed. Y. Pomeranz), Chap.
4, pp. 139-242, Am. Assoc. Cer, Chern. Inc., St. Paul, MN.
Stauffer, C.E. (1996). Emulsifiers for the food industry, in Bailey's Industrial Oil and
Fat Products, 5th ed., Vol. 3, pp. 483-516, Wiley, New York.
Tenney, R.J. (1978). Dough conditioners/bread softeners, Baker's Digest, 52 (4), 24.
Tsen, C.C., et al. (1973). High protein cookies I. Effect of soy fortification and surfactants, Baker's Digest, 47(4); 34-39.
Weiss, T.J. (1983). Food Oils and Their Uses, AVI, Westport, CT.
Wooten, J.C., et al. (1967). The role of emulsifiers in the incorporation of air into layer
cake batter systems, Ger. Chem., 44, 333.

NINE

Emulsifiers in Confectionery
Mark Weyland

9.1

Introduction

The use of emulsifiers in both chocolate confectionery and sugar confectionery


has been established for many years. They are useful functional additives
without which many candy products would be impossible to make. In this
chapter, an outline of the range of emulsifier functionalities will be presented,
but since it is a very diverse field, it is not possible to cover every application
or use. A bibliography is provided for further study, which is recommended.
Emulsifiers act in multiphase confectionery systems in two main ways. The
first is as an emulsifying agent to enable two distinct phases to be combined in
a stable quasi-homogeneous state for an indefinite length of time. The two or
more phases involved in most food products may be selected from a list that includes oil, water, protein, carbohydrates, and air. These phases have a natural
tendency to repel each other and form discrete layers within a food product. In
most food products this tendency to separate into phases is undesirable and
must be controlled by a suitable blend of processing techniques and carefully
selected emulsifying agents. Furthermore, even if the food product is satisfactory at the time of production, it must still withstand the rigors of distribution
and storage on the shelf, such that at the point of consumption by the con-

235

236

Food Emulsifiers and Their Applications

sumer the product has an acceptable taste, appearance, and texture. These
qualities in modern food products are often critically dependent on the type
and level of emulsifiers used in the product.
The second function of an emulsifier is often to act as a surfactant. In these
cases the role of the emulsifier is to modify the behavior of the continuous
phase of a food product so as to bring about a specific effect or benefit. The
most common example of this in confectionery is the use of lecithin in chocolate to reduce the viscosity of the product and improve the ease of handling
and processability.
Many of the classes of emulsifiers described in this book have also found
their way into confectionery products. These include lecithin and modified
lecithins such as YN and phosphated monoglycerides, glycerol monostearate,
polyglycerol esters including polyglycerol polyricinoleate (PGPR), sorbitan esters, polysorbates, lactic acid and tartaric acid derivatives of monoglycerides,
acetylated monoglycerides, sucrose esters, and propylene glycol monoesters.
All these compounds have a common feature that makes them suitable as
emulsifying agents: they are ambiphilic, possessing both lipophilic and hydrophilic properties. The nature of this property is often expressed as hydrophilic/lipophilic balance or HLB. The HLB number is an indication of the
properties of an emulsifier and is usually on a scale of 0 to 20. An emulsifier
with a low HLB will tend to be more oil-like and will therefore have a greater
affinity for the oil phase of a confectionery product. Lecithin, for example, has
an HLB of 4 and has a great affinity for the oil phase in chocolate. Polysorbate
60, by contrast, has an HLB of 15 and is quite soluble in water and therefore
has an affinity for the syrup phase in toffees and caramels.
It is often the case in food products, and in confectionery too, that a combination of two emulsifiers in a recipe or formula containing two distinct phases
will result in the longer lasting and more uniform product. In these cases, combinations oflow- and high-HLB emulsifiers give the best results.
In the remainder of this chapter, a number of the more common confectionery types using emulsifiers are described, along with a review of the available knowledge relating to the most optimal emulsifier types and their benefits.

9.2

Emulsifiers in Chocolate and Compound Coatings

The use of emulsifiers in chocolate and compound coatings is perhaps the best
documented in the literature of any of the applications of these ingredients in

Emulsifiers in Confectionery

237

confectionery. The recent changes in the Standard of Identity for chocolate


products (1993) has led to the possibility of using any safe and suitable foodgrade emulsifier in chocolate. A number of emulsifiers used in compound or
vegetable fat coatings with advantage can now also be used in chocolate. Other
emulsifiers not available for use in the United States at this time are used in
the EC and can bring particular benefits to the chocolate and compound coating user. These will also be described later, but first there follows a description
of the most widely used emulsifier, soy lecithin.

9.2.1

Lecithin

Chocolate and compound coatings are dispersions of solid particles in a continuous fat phase. The solid particles are composed of sugar granules, milk
solids, and cocoa solids. In chocolate the fat is cocoa butter and comes directly
from the crushing of cocoa nibs, but in compound coatings the fat comes from
vegetable oils added to the formula to give the same fat content as chocolate
overall. Moisture is also present in the coating, introduced indirectly via the
sugar or other solid ingredients. It is the presence of these solid particles and
moisture that causes chocolate and compound coatings to deviate from true
Newtonian viscosity behavior. The solid particles tend to abrade each other,
and the resultant internal friction causes the viscosity of the material to vary
according to the applied shear stress.
Viscosity is very important when considering how chocolate and compound
coatings are used, because they always have to flow to either fill a mold without
defects or air bubbles or cover a candy piece with a thin, even coat. These
properties are controlled almost completely by the rheological behavior of the
coating, which in turn is dependent on the nature of the continuous liquid
phase, that is, the fat and fat-soluble ingredients. Considering the flow properties of molten chocolate, the two most important parameters are plastic viscosity and yield value. "Plastic viscosity" is defined as the force required to keep
liquid chocolate flowing once it has started moving, whereas "yield value" is
the force required to start the mass of liquid chocolate moving. Both plastic
viscosity and yield value can be decreased by the use of specific surfactants,
and this enables the chocolate manufacturer to have greater control of cocoa
butter levels. Plastic viscosity and yield value are often combined in a value
called "apparent viscosity," but it is important to understand that chocolates
with equal apparent viscosities can have different yield values and different
plastic viscosities.

238

Food Emulsifiers and Their Applications

Coatings can always be made more fluid for better control by adding more
cocoa butter or vegetable fat to the mix, but since these are the more costly ingredients in coatings, this is often an unattractive solution. Better by far is to
add a surfactant like lecithin to reduce coating viscosity. It is possible to add
0.5% lecithin to a coating to have the same viscosity reduction effect as the
addition of 5% cocoa butter or vegetable fat (Minifie, 1980). Lecithin allows
coating users to operate efficiently at much lower fat contents than would otherwise be the case (Figure 9-1).
Lecithin is commercially extracted from the soybean by solvent extraction
and precipitation. It is a light brown fluid that contains approximately 65%
acetone-insoluble phosphatides and 35% soybean oil. An oil-free product is
also available that is in the form of granules. The chemical composition of soy
lecithin is as follows (Minifie, 1980):

Soybean oil
Phosphatidyl choline
Phosphatidyl ethanolamine
Phosphatidyl inositol
Other phosphatides and polar lipids
Carbohydrates, e.g., sterols

35%

18

15
11

9
12

39
38
L.

Q)

~
::I

.0

co

37
36

(.)

35

'#-

34

(.)

33
32

1\

""
0.1

0.2

..........

0.3

['...

0.4

0.5

0.6

0.7

% lecithin (soya)
Figure 9.1 Effect of addition of lecithin on the fat content in dark enrobing chocolate.
Curve represents chocolate formulas of equal viscosity. (Minifie, 1980.)

Emulsifiers in Confectionery

239

A structure for lecithin is given in Figure 9.2.


Lecithin exhibits both lipophilic and hydrophilic properties. A possible explanation of the mechanism by which lecithin reduces intraparticle friction is offered by Harris (1968). Moisture present in chocolate and compound coatings
adher-es to the surface of sugar particles and gives them a syrupy, tacky surface
that in tum increases friction between the sugar grains. When lecithin is introduced the hydrophilic functional group in lecithin attaches itself to the sugar
surface while the lipophilic group is left to project out into the surrounding oil
phase. This enables the particles to slip more easily over each other, reducing
the viscosity. This effect is demonstrated in Figure 9.3, in which the viscosity-reducing effect of lecithin is seen only where sugar is present in the formula.
Lecithin is usually added late in the chocolate or compound-making
process since it can be adsorbed by cocoa particles during grinding and mixing, thereby losing its effectiveness. In some cases a small amount of lecithin
is added to the mixed ingredients prior to roller refining to aid in the grinding
process, but the remaining portion is added just before the end of the conching
process. This provides the maximum liquification of the chocolate or compound coating at minimum fat content.
Lecithin also has the benefit of protecting coatings against moisture invasion and sugar granulation, which may occur at above 60C when stored in
bulk form. Lecithin used in excessive amounts will not further reduce viscosity
of chocolate or compound coatings but will produce certain negative effects
such as softening of chocolate and increase of crystallization time (Jeffrey,
1991). This is because the chemical structure of lecithin is very different from
cocoa butter or vegetable fats, and this can interfere with the crystallization

CH 2-o-c-R

?!
CH-0-C-R

lipophile

II
I

hydrophilic

CH 2-o-P-O-cH 2-CH 2-N-(CH 3 ) 3


OH

Figure 9.2

II

OH

Chemical structure of soy lecithin. (Woods, 1976.)

240

Food Emulsifiers and Their Applications

100

[',_

1'---

90

80

r--__
c---

..

50

>"'

40
30
20

10

1---

--

!\

--: 60
~
>.

---- - -

~--

70

"'utJ

~Cocoa/Fat

\\

\l

""
0 .1

'

- - .........

t---- -

Chocolate

I'-.

1'--.:_ugor I Fat

0 .2

0 .3

0.4

0.5

0 .6

0 .7

"I Lecithin (Soya)

Figure 9.3 Influence of lecithin on viscosity measured using a Redwood viscometer.


(Minifie, 1980.) (In this viscosity measurement method, the time taken for a known
amount of chocolate to drain through a hole in a cup is measured using a stopwatch.
This value in seconds gives an empirical value for chocolate consistency.)

process in the fat phase. Levels of over l% in typical 30% fat coatings should
not be used.

9.2.2 Synthetic Lecithin


These products are used in chocolate and compound coatings outside the
United States and are made by reacting mono-, di-, and triglycerides of partially hydrogenated rapeseed/canola oil or other liquid vegetable oil with phosphorus pentoxide to produce phosphatidic acids. Neutralization with ammonia
or caustic soda results in an ammonium or sodium salt. These surfactants are
often given the name synthetic lecithins or sometimes YN lecithin. They have

Emulsifiers in Confectionery

241

a neutral flavor and can be used at higher dosage levels than natural lecithin
without the negative impact on viscosity (Bonekamp-Nasser, 1992; Klienert,
1976; Nakanishi, 1971). See Figure 9.4.
YN is also claimed to reduce the thickening of chocolate and compound
coatings due to moisture and overheating (Bradford, 1976). A comparison between lecithin at 0.3%, YN at 0.3%, and cocoa butter added at 5% to chocolate gave the same similar overall viscosity readings (see Figure 9.5), but a
calculation of the Casson yield values showed that YN produced significantly
lower values than with the other systems. The viscosity reducing effect of YN
is reportedly less in milk-free coatings than with milk coatings (Klienert,
1976; Hogenbirk, 1989). Milk coatings have generally higher viscosities than
milk-free coatings due to the effect of milk solids/fat/emulsifier interactions.

150
140
130
120

~ 1 RP.M.

90

tl

ao

0"'

RPM. : Rate of shear

\\

\\

100

--

\"''

""

'

''

60

50

'-..5

0
0

40

L.

cD

--- -

..............
.....

..........

20

20 R.P.M.

...

0
0.1

0.2
0

/o

--.. _

... -r--

~--

10

Figure 9.4

... ... ...

---

r--

..........

R.P.M.

'<"..... r::--..........

30

r--------

'~

--

?- 70

"'u0
>"'
"0
u

Soya lecithin

---YN

~\

110

....

~\

0.3

.... --

--- 1----------04

0.5

Lee ith 1 n

Brookfield viscosity measurements of milk chocolate at three levels of


shear. Comparison of effeet of soya lecithin and YN lecithin. (Minifie, 1980.)

242

Food Emulsifiers and Their Applications

700

c:;-

5% cocoa butter

E
..s?.
Ul

600

0.3% lecithin

Q)

c:

>

500

~
Ul
Ul
Q)

--+-

0.3% YN

__.._

400

....
....

+'

Ul

300

ctl

Q)

..c:

(/)

200

Figure 9.5

10

20

30

Shear rate (s-1)

40

50

Viscosity plot comparing lecithin, YN, and cocoa butter. (Bradford, 1976.)

These interactions result in higher viscosities compared to coatings containing


only cocoa solids and sugar for surfactant adsorption. Details of how these interactions occur are absent from the literature.

9.2.3

Polyglycerol Polyricinoleate (PGPR)

PGPR is a surfactant used in the chocolate and compound industries in


Europe and other parts of the world, excluding the United States. It has a
unique role to play in modifying the viscosity behavior of chocolate coatings. It
is made by reacting polyglycerol with castor oil fatty acids under vacuum. The
resultant material is a colorless, free-flowing fluid with little or no odor. PGPR
is also claimed to he a moisture scavenger in chocolate and compound coatings, preventing thickening of coatings over time (Application Notes Admul
WOL, Quest International).
Its chemical structure is, in general form,

where R= H or a fatty acyl group derived from poly condensed ricinoleic acid
and n is the degree of polymerization of glycerol.

Emulsifiers in Confectionery

243

A number of studies have been published that compare the effects of PGPR
with lecithin and YN. Most conclude that PGPR, when added to chocolate or
compound coatings at 0.5% or less, can reduce the coating yield value to almost zero (Application Notes Admul WOL, Quest International; Bamford,
1970). The practical benefit of such a feature is that in a chocolate bar molding
operation, PGPR addition would allow the chocolate to flow easily into even
complicated mold shapes without entrapping air bubbles and also to flow
around inclusions. Furthermore, the opportunity exists to reduce the fat content of the chocolate as well as the coat of chocolate formulations.
A typical comparison of lecithin and PGPR additions to a milk and to a
dark chocolate using a Rotovisco viscometer is given in Tables 9.1 and 9.2
(Application Notes Admul WOL, Quest International).
The values in Tables 9.1 and 9.2 indicate that in milk chocolate it is possible to reduce the yield value to almost zero. The presence of lecithin in the
emulsifier mix allows the plastic viscosity to be minimized, and this blend pro-

Table 9.1

Casson plastic viscosities and yield values of a milk chocolate when


cocoa butter, lecithin, and PGPR are added
Casson plastic
viscosity (poise)

Casson yield value


(dynes/cm2 )

2.0
4.0
5.0

45
29.8
26.5
16.3
15.3

97
62
58
58

Lecithin

0.05
0.1
0.2
0.4

30.0
26.7
20.0
15.6

79
54
40
37

PGPR

0.075
0.175
0.3
0.5
0.6

30.0
29.2
26.8
30.5
32.0

86
38.5
22
2.5
2.0

Lecithin + PGPR

0.1
0.2
0.3

14.1
13.4
12.7

34
32
29

Addition

Amount

Cocoa butter

0.0

l.O

llO

244

Food Emulsifiers and Their Applications

Table 9.2 Casson plastic viscosities and yield values of a dark chocolate when cocoa butter, lecithin and PGPR are added
Addition

Amount

Casson plastic
viscosity (poise)

Casson yield
value (dynes/cm2)

Lecithin

0.3
0.7
0.97
1.3

18.5
17.1
14.4
12.4

155
221
297
285

PGPR

0.0
0.1
0.2
0.5
1.0

12.9
12.5
14.8
14.9
15.9

199
151
82
13
0

vides the optimum solution. In dark or semisweet chocolate the effect of PGPR
on plastic viscosity is small, while it can reduce yield values to very low values
at 0.5% addition. The mechanism by which PGPR affects yield value is not
understood but its practical benefits are widely reported.
PGPR is also claimed to be advantageously used in ice cream coatings since
it allows low apparent viscosities in the presence of low levels of moisture
(Bamford, 1970). Also claimed is PGPR's beneficial effect on fat-phase crystallization leading to easier tempering, improved texture, and longer shelflife of
coatings (Application Notes Admul WOL, Quest International.). The author has
found that the viscosity-reducing properties of PGPR does lead to significantly
reduced viscosity at temper and a level of temper, as measured by a tempermeter, which is easier to maintain over long periods in an enrober without significant recirculation of chocolate via melt-out and retempering circuits.
PGPR's most recognized benefit remains that of fat reduction, and manufacturers claim that a blend of 0.5% lecithin and 0.2% PGPR allows cocoa butter
reductions of approximately 8%.

9.3 Antibloom Agents in Chocolate


and Compound Coatings
There are many references in the literature to uses of emulsifiers as additives
to influence the physical appearance and texture of chocolate and compound
coatings. One of the main problems to be overcome when making cocoa but-

Emulsifiers in Confectionery

245

ter-based chocolate is to ensure that the cocoa butter crystallizes in the correct crystal form or polymorph. Cocoa butter has several different polymorphic
forms that have melting points ranging from 17 to 35C. The forms are represented by the greek letters y, a, pt, and~ As the polymorphic form increases
in stability it also increases in melting point. To make chocolate in the familiar
glossy, fast-melting form with good snap, it is necessary to crystallize the cocoa
butter in the highest melting polymorph, which is ~ This form of cocoa butter
is also needed to ensure good contraction in molded products and the long
bloom-free shelf life expected for good quality chocolate goods. Bloom is the
phenomenon in which liquid fat is forced to the surface of a product and spontaneously crystallizes there as a grayish layer that looks to the uninformed consumer as if the product has gone moldy. The liquid fat is forced out to the
surface in this way by the following mechanisms:
l. Chocolate is not preconditioned (tempered) correctly such that insufficient concentration of seeds in the ~ form is present in the crystallizing
chocolate mass. This leads to a higher level of less stable Wforms in the

chocolate mass that later transform to the more stable ~ form. This
transformation causes the chocolate coating or bar to contract and
squeeze liquid fat to the surface. Chocolate contains liquid fat even at
room temperature, where cocoa butter attains a maximum solid fat content of approximately 85%. This liquid fat at the surface crystallizes in
an uncontrolled fashion and is a mixture of ~' W, and even possibly
some a forms.
2. Chocolate is tempered correctly, but in storage or distribution the product is subjected to wide temperature variations, resulting in partial
melting and resolidification of the chocolate. Under these conditions
uncontrolled recrystallization takes place and extensive bloom can occur. This kind of change is often referred to as "heat damage" and the
product is classified as not heat resistant.
3. In molded bars that contain peanuts or other nutmeats as solid inclusions, or in enrobed products that have centers containing quantities of
soft vegetable oil or dairy butter oil, this oil can "migrate" from the center to the chocolate shell. The soft oil will cause the chocolate to become
soft and cocoa butter will dissolve. This will cause severe damage to the
product due to physical handling prior to consumption or due to discoloration and bloom of the chocolate shell, which will now be far more
heat-sensitive.

246

Food Emulsifiers and Their Applications

4. Long-term changes in cocoa butter crystal structure via ~(V) to ~(VI)


transitions can also be a cause of bloom in some cases, although this
may not be as common as the mechanisms above. In this scenario cocoa
butter in the stable ~ state can exist in two forms, given the nomenclature V and VI. At the time of manufacture only the form V can he
formed. Form VI has a slightly lower energy than form V, but the transformation from one to the other takes place at a very slow rate because
there is a significant kinetic harrier to he overcome. This bloom mechanism occurs only in solid dark (milk-fat-free) chocolate bars. Milk fat
present at low levels (2%) in chocolate will prevent this type of bloom.
(See also the reference to STS as a bloom inhibitor in Section 9.3.1.)
In all the cases above, the negative impact of uncontrolled crystallization is
discoloration and bloom. This phenomenon is also seen in compound coatings
based on other vegetable fats, hut since they are not polymorphic in the same
way as cocoa butter, the mechanism of bloom formation is different. Problems
of discoloration tend to stem from types 2, 3, and 4 above rather than type l.
The final result is, however, the same, with an unattractive dull finish becoming apparent on the coating surface.
Emulsifiers can have a role in helping control the rate of crystallization of
cocoa butter and other vegetable hard butters both at the time of production
and during subsequent storage and distribution. This in turn can promote
bloom prevention.

9.3.1

Sorbitan Tristearate (STS)

STS is an emulsifier often associated with bloom prevention; it is claimed that


when added to chocolate in the liquid state at 2% it slows down the crystallization rate of cocoa butter, thereby reducing the concentration of the most unstable

a form. The more stable

Wform is still produced hut this transforms into

the~ form thus deterring bloom (Anon., l99la). In this way STS behaves as a

crystal modifier.
The situation is further complicated, however, because there exists not one
hut two ~ forms of cocoa butter, usually referred to as forms V and VI. In this
classification system the Wform is called form IV and transitions from form IV
to form V, the stable form, occur via the liquid phase only. The transformation
of form V to form VI takes place only via the solid phase and can take many
months to occur. Also, identification of form VI is difficult and requires specialized x-ray diffraction techniques. However, this ~transformation is also as-

Emulsifiers in Confectionery

24 7

sociated with bloom in solid chocolate that has been well tempered. Work done
by Garti et al. (1986) has indicated that STS is particularly effective at blocking this V to VI transformation and, hence, preventing bloom even after extensive temperature cycling between 20 and 30C.
Other emulsifiers studied by Garti included sorbitan monostearate and
Polysorbate 60, but these were only half as effective as STS. STS is a highmelting-point emulsifier (approximately 35C} whose structure is more closely
related to cocoa butter triglycerides than to most other emulsifier types. It is
speculated that it is due to this similarity that STS co-crystallizes with cocoa
butter from the melt and, due to its rigid structure, binds the lattice in form V.
Other more liquid or less triglyceride-like emulsifiers tend to depress the melt
point of crystallized cocoa butter, increasing liquidity and promoting form IV
to V transformations in preference.
This is presumably why STS is a more effective antibloom agent in solid
chocolate than in enrobed chocolate items, where soft center oils can soften
cocoa butter crystals via migration and lead to type IV-V bloom occurring (see
type 3 bloom previously mentioned). Krog (1987), however, claims that STS
locks fats in the less stable Wform and prevents the transformation to ~
Berger (1990) also claims that STS performs well as a bloom inhibitor or gloss
enhancer in palm kernel oil-based compound coatings used to enrobe cakes

by stabilizing the Wform of the vegetable fat, a situation also observed by the
author in several practical cases using lauric coating fats but with much less
reliability when using domestic fats such as soybean or cottonseed-based coating fats. Such products tend to have longer bloom-free shelf lives in many
cases so that the need for antibloom additives is not so imperative.
The use of STS in food in the United States is limited to chocolate and compound coatings for which there is a petition pending for acceptance by FDA as
a food additive. This uncertain status has not deterred United States food companies from enjoying the benefits that STS can bring to coating appearance.
STS is, however, more widely accepted as an additive in EC countries.

9.3.2

Sorbitan Monostearate (SMS) and Polysorbate 60

SMS and polysorbate 60 [also known as polyoxyethylene (Player, 1986) sorbitan monostearate] are also used as antibloom agents, especially in compound
coatings based on vegetable butters. They are not as effective as STS, but they
have the advantage of being already accepted by FDA as food-grade emulsifiers. They are usually used in combination, where the SMS acts as a crystal

248

Food Emulsifiers and Their Applications

modifier and the polysorbates act as hydrophilic agents to improve emulsification with saliva and aid flavor release (Dziezak, 1988; Lees, 1975). SMS can
also be used at high levels in coatings to increase heat resistance due to its
high melt point, 54C; unfortunately, the addition of SMS also causes the coating to become waxy.
Up to 1% of this combination can be added to coatings to improve initial
gloss and bloom resistance. The optimum ratio of SMS to polysorbate 60 is
60:40 (Woods, 1976). These emulsifiers are claimed to function by forming
monomolecular layers of emulsifier on the surface of sugar and cocoa particles,
thereby inhibiting the capillary action that causes liquid fat to migrate to the
surface and cause bloom. Lecithin is still needed in these systems to control
coating viscosity and reduce fat content.
In another nomenclature system, SMS is also called Span 60 and polysorbate 60 is called Tween 60 (Lang, 1974). Spans and Tweens are also claimed to
reduce the rate of fat crystallization; therefore, to develop proper crystal size a
suitable tempering system needs to be employed. They are employed in both
chocolate and compound coatings and may be used with advantage iffast crystallization of the coating would be disadvantageous.

9.4

Other Emulsifiers Used in Coatings

Mono- and diglycerides are also used as additives to chocolate and compound
coatings, often as their purified or distilled forms. They can act as seeding
agents especially when in high-melting-point forms such as glycerol monostearate (GMS). They are more commonly used as antibloom agents in laurictype palm kernel oil compound coating to extend useful shelf life. A typical
usage level would be 0.5%. Berger (1990) claims good results in hydrogenated
palm kernel oil coatings when using glyceryllacto palmitate at 1 to 5% as a
gloss improver; the application was as a coating for a baked product. Moran
(1969) found that a polyglycerol ester of stearic acid reduced the viscosity of
fat sugar systems better than lecithin as well as retarded crystallization, improved gloss, and better demolding.
Lactic acid esters of monoglycerides have also been used to control gloss in
compound coatings (Hogenbirk, 1989; Dziezak, 1988) and to improve demolding performance (Anon., 1991b). Woods (1976) describes the use of triglycerol
monooleate in compound coating chocolate to improve initial gloss and gloss
retention, and triglycerol monostearate as a whipping agent to aerate coatings

Emulsifiers in Confectionery

249

giving them a lighter texture for filling applications. Herzing (1982) describes
in detail the types of polyglycerol esters needed to optimize the glossy properties of lauric and nonlauric compound coatings: triglycerol monostearate, octaglycerol monostearate, and octaglycerol monooleate. These emulsifiers are
added to the coating fat at up to 6% by weight.
Polyglycerol esters have also been claimed to speed up the setting time of
chocolate panning coatings when used at levels of 0.4 to 0.6% (Player, 1986).
Hogenbirk (1989) has found advantages of viscosity reduction to some degree
with examples of mono- and diglycerides, diacetyl tartaric acid esters of monoglycerides (DATEM), acetylated monoglycerides, and propylene glycol monoesters. Musser (1980) has published results showing the benefits of adding
up to 1.5% DATEM to chocolate and compound coatings to modify viscosity
properties and to improve the rate of crystallization of coating fat phases. In a
series of experiments, Musser found that the addition of DATEM to fully
lecithinated milk and dark chocolates, and dark sweet coatings, could further
reduce the viscosity of the coatings as measured by a Brookfield viscometer. At
the same time Musser found that the DATEM was acting as a seeding agent in
chocolate systems, improving the speed of crystallization and resulting in a
finer grain and better gloss in molded bars. Musser's conclusions relative to the
effect of DATEM on viscosity is supported by the author's own published study
on chocolate viscosity and emulsifiers (Weyland, 1994).

9.5

Emulsifiers in Nonchocolate Confectionery

Unlike in chocolate and compound coatings, the continuous phase of sugar


confectionery is not oil or fat but is either sugar or a sugar syrup (in this case
"sugar" means any nutritive carbohydrate sweetener). For this reason the role
of an emulsifier in sugar confectionery is to enable small quantities of
lipophilic material to be finely dispersed within a sugar matrix to achieve a desired effect. This effect may involve the dispersion of color, flavor, or some
other fat-soluble ingredient, or the direct physical interaction of the emulsifier
with the sugar phase to achieve the desired textural properties.
A major factor in consumer acceptance is the mouthfeel of a confection.
Vegetable fats and emulsifiers are used to improve texture and lubricate the
product to achieve better chewing characteristics. A well-chosen surface-active
agent can improve this aspect as well as slow down the release of added flavorings. They will affect the viscosity characteristics of the sweet and influence the

250

Food Emulsifiers and Their Applications

crystal shape present in grained confections. The improvement in fat dispersion


throughout the confection will slow the rate at which the ingredient becomes rancid as the amount presented or migrating to the surface is lessened.
For the purposes of this review I will divide the use of emulsifiers into the
following categories: (i) chewing gum; (ii) caramels, toffee, and fudge; and (iii)
jellies and gums.

9.5.1

Chewing Gum

Chewing gums contain fats and emulsifiers that act as softening agents or plasticizers to the gum base. In this role, emulsifiers can also act as carriers for
colors and flavor aiding in the dispersion of these important ingredients within
the gum base. Up to 1% lecithin can be used to soften chewing gum to the desired consistency (Patel et al., 1989) and can be hydrated or mixed with a vegetable oil or suitable fatty emulsifier such as mono- and diglycerides, to aid in
dispersion within the chewing gums. Chewing gums prepared in this way have
the desirable soft, chewy properties popular in today's top products.
Other emulsifiers are also used in chewing gum to provide suitable textural
and antistick properties to the chewing gum base; and these include monoand diglycerides, glyceryl lacto palmitate, sorbitan monostearate, triglycerol
monostearate, triglycerol monoshortening, and polysorbates 60, 65, and 80.
Lecithin is also used to provide a protective coating to chewing gum pieces
prior to a hard panning process (Dave et al., 1991). Normally, only hard chewing
gums can be hard panned in this way, but by using a hydrated lecithin coating it
is possible to candy coat the gum and then allow the lecithin to soften the chewing gum in storage prior to consumption. By this technique the emulsifier coating, when set, forms a suitable base for syrup-based candy coatings.

9.5.2

Caramels, Toffee, and Fudge

Emulsifiers acting as surface-active agents increase the resistance of caramels


and similar fat-containing confections to stick on cutting knives or to oil out due
to manipulation in the process. The use of glycerol monostearate (GMS) and similar products such as mono- and diglycerides do produce an improvement in lowfat products such as in toffee and nougat. It is also claimed that the addition of
these agents can help bind the oil and oil-based flavoring into a confection, reducing sweating and problems with rancidity. One property of emulsifiers that is
particularly important is the improvement in emulsification in high-fat-content

Emulsifiers in Confectionery

251

confections such as butterscotch and caramels. During the manufacturing


process, a high-sugar-content syrup is produced in which fat globules are dispersed. The finer these globules are the better will be the eating qualities.
Consumer acceptance is based on flavor, appearance, texture, and chew.
The last two factors are influenced directly by the number and size range of fat
globules present in the product. The lower the size and the more even the dispersion of fat globules, the smoother the confection. Fat-containing products of
this type exhibit non-Newtonian flow with a yield stress, which means that it is
necessary to apply a sufficient level of stress to cause flow. Fat makes a minor
contribution to the viscosity in relation to that due to the milk protein, but its
presence is extremely important for lubrication. The higher the level of water
left in a confection of this type, the higher the danger of stickiness and graining. GMS is a highly effective emulsifier to ensure maximum fat dispersion and
minimum stickiness, but glycerol monooleate and lecithin are also quite effective. A usage level of 0.25 to 0. 7% is normal.
Fudges are fondants that contain milk protein and fat having a flavor characteristic of toffee and caramel. Its texture can vary widely from a hard product
often sold as a cut tablet to a soft form used in starch-molded products. In
fudgemaking, an emulsified milk phase is prepared consisting of sweetened
condensed milk, vegetable fat such as hardened palm kernel oil, and an emulsifier, usually GMS. The fat is emulsified into the syrup solution in a highspeed mixer at 120 to l40F. Without GMS present the fat is more likely to
separate during the subsequent cooking and working process or during the
storage life of the fudge. This emulsified milk phase is added to a sugar/corn
syrup mix and cooked to the desired moisture content of 8 to 15%. During this
cooking process the characteristic Maillard or caramelized flavors develop.
Finally, fondant is added during the cooling stage to provide the correct
level of sugar seeds to make the desired sugar crystal size in the fudge. The
fudge is then sheeted, deposited, or extruded.

9.5.3

Jellies and Gums

Some emulsifiers and surface-active agents such as GMS and saturated


ethoxylated monoglycerides or polyglycerate 60 are adsorbed onto starch granules. This property can be used to modify the texture of starch-based sugar
confectionery. Gel formation in starch-based jellies and gums is mainly due to
the water-soluble fraction of starch, the amylose. Interaction between amylose

252

Food Emulsifiers and Their Applications

and emulsifiers creates a water-insoluble helical complex and creates an irreversible textural effect. This interaction was quantified by Krog (1977) using
the amylose-complexing index or ACI. The ACI is defined as the percentage of
amylose precipitated at 60C after l hour and after reacting 5 mg of the emulsifier with 100 mg of amylose in solution. See Table 9.3.

Table 9.3

ACI values of some food emulsifiers

Glycerol monostearate (85%)


Glycerol monooleate {45%)
Mono- and diglycerides (50% monoester)
DATEM
Sorbitan monostearate
Lecithin
Polysorbate 60
Acetylated monoglycerides

87
35
42
49
18
16
32
0

To be an active amylose-complexing agent an emulsifier must have a high


level of saturated monoglycerides and some degree of water dispersability. An
example of the use of emulsifiers in starch-based confectionery is in the making
of Turkish delight, where it is possible to use emulsifiers with high ACI values
(GMS) to avoid pastiness or cheesiness. Usage levels are typically 0.025%.

9.6

Processing Aids

Emulsifiers are sometimes used in small amounts in confectionery products either to control aeration or to prevent product sticking to machinery and packaging. They can also be used to displace starch from starch-molded jellies and
gums and provide a shiny attractive appearance as well as a barrier to degradation from atmospheric oxygen and moisture.
Aeration of protein systems containing small amounts of fat, such as
nougats, can be facilitated by the addition of triglycerol monostearate. Liquid
fat or lipophilic emulsifiers such as GMS or acetylated monoglycerides usually
tend to destabilize forms and cause deaeration.
Emulsifiers are also useful release agents providing barrier properties between product and molds, tables, metal, conveyor belts, utensils, and machin-

Emulsifiers in Confectionery

253

ery, especially on cooling. Release agents must be food-grade materials and


have high stability to resist oxidation and hydrolysis.
Acetylated monoglycerides are used as release agents or as oiling and polishing agents because they form stable films on the surface of confectionery
items. They have a-crystalline stability, plastic, nongreasy texture, and neutrality of flavor, color, and odor. They reduce shrinkage, harden through moisture loss, and prevent fat degradation and mold growth. They retain moisture
and other desirable properties of the foodstuff and prevent contamination by
moisture or dust. They are usually applied directly to the confectionery product by spraying. Melting points used are in the range 30 to 46C. Typical applications include nuts and dried fruits and certain panned confectionery
items. Lower melting-point forms (l0C) can be sprayed directly onto conveyers and molds to release goods with high sugar contents such as fondant
creams and jellies.
Another release agent used often on chocolate-enrobing tunnels is a mixture of lecithin and cocoa butter. This is sprayed onto the band before the
candy center is deposited to ensure clean separation of the centers from the
band prior to chocolate-enrobing.

References
Anon. (199la). Confectionery Production, 57(2), 136-7, 140.
- - - (l991b). Confectionery Production, 57(6), 451-2.
Bamford, H.F., eta!. (1970). Rev. Int. Choc. (RIC), 25, 6.
Berger, K.G. (1990). World Conference on Oleochemicals into the 21st century, AOCS,
Champaign, IL, pp. 288-91.
Bonekamp-Nasser, A. (1992). Confectionery Production, 58(1), 66, 68.
Bradford, L. (l976).Int. Flavors Food Addit., 7(4), 177-9.
Dave, ].C., eta!. (1991). U.S. Patent 5,135,761, March 28.
Dziezak, J.D. (1988). Food Techno!., 42(10), 171-86.
Garti, N., eta!. (1986). ]. Assoc. Off. Chem. Soc., 63(2) 230-236.
Gregory, D.H. (1982). Confectionery Production, 48(10), 437-9.
Harris, T.L. (1968). Surface Active Lipids in Foods, Monograph No. 32, Society of
Chemical Industry, England.
Herzing, A.G., eta!. (1982). U.S. Patent 4,464,411, November 5.
Hogenbirk, G. (1989). Confectionery Production, 55(1), 82-3.
Jeffrey, M.S. (1991). Manufacturing Confectioner, 71(6), 76-82.
Klienert, J. (1976). Rheol. Texture Food Qual., pp. 445-73, AVI, Westport, CT.
Krog, N. (1977). ]. Assoc. Off. Chem. Soc., 54(3), 124--31.
Lang, M. (1974). Confectionery Manufacturing and Marketing, 11(2), 3-5, 13.
Lees, R. (1975). Confectionery Production, 41(6), 296,298, 304.

254

Food Emulsifiers and Their Applications

Minifie, B.W. (1980). Manufacturing Confectioner, 60(40), 47-50.


Moran, D.P.J. (1969). Rev. Int. Choc. (RIC), 24,12.
Musser, J.C. (1980). 34th PMCA Production Conference, Lancaster, PA.
Nakanishi, Y. (1971). Rev. Int. Choc. (RIC), 26, 8.
Patel, M.M. et al. (1989). U.S. Patent 5,041,293, December 28.
Player, K. (1986). Manufacturing Confectioner, 66(10), 61-5.
Weyland, M. (1994). Manufacturing Confectioner, 74(5).
Woods, L.C. (1976). Gordian, 76(2), 53-7.

TEN
Margarines and Spreads
Eric Flack

10.1

Introduction

The invention of margarine in the 1860s is attributed to the French chemist


Hippolyte Mege Mouries, in response to a competitive challenge initiated by
the French Government under Napoleon III for a less expensive and less perishable substitute for butter.
Mege had already undertaken research in this field at the Imperial farms,
where he had observed that although underfed or hungry cows lost weight, they
still produced milk, though of lower than normal yield, and that the milk contained fat. Thus, he deduced that milk fat was derived from normal body fat,
i.e., tallow. However, since tallow itself does not possess the melting property
of milk fat, he construed that through some metabolic process, a fractionation
of the fat occurred, the lower melting fractions of which were transported to the
mammary glands of the udder whereby, through the enzymatic action of
pepsin, it became transformed to butter fat and dispersed as an emulsion in
the milk plasma, while the harder fractions were utilized by the animal as a
source of energy (Andersen and Williams, 1954).
Having arrived at these conclusions, Mege set out to imitate this natural
process by carefully rendering fresh tallow at body heat (about 45C) using ar-

255

256

Food Emulsifiers and Their Applications

tificial gastric juices to facilitate separation of tissue from pure fat and then,
following crystallization at 25 to 30C, extracting under pressure about 60% of
a soft semifluid fraction-oleomargarine-and about 40% of a hard white
fat-oleostearine. The softer fraction had a melting point similar to butter fat,
a pleasant flavor not unlike melted butter fat, a pale yellow color, and could
easily be plasticized, and although different in fatty acid composition from butter fat, it offered a useful basis for the production of a substitute. He assumed,
therefore, that this new butter fat substitute consisted of glycerides of margaric
and oleic acids (the former then considered to be a C17 homolog of the saturated fatty acids, although now known to be a eutectic mixture of palmitic and
stearic acids) that solidified in crystals having a pearly luster. The name margarine was derived from the Greek word "margarites," meaning pearl and thus
is pronounced with a hard "g" according to its derivation.
Mege's process was to take a proportion of the soft fat with appropriate
quantities of milk and water, into which a small amount of udder extract was
stirred. Following agitation, a stable emulsion similar to thick butter cream
formed that, on further churning, thickened to resemble butter. While this
process may now sound rather rudimentary, it nonetheless comprised all the
elements of margarine production as we know them today.

10.2

Early Development

Mege took out patents in France and England, but the next major development
took place in Holland when, in 1871 in the absence of patent law in Holland at
that time, he sold his invention to the butter merchants Johannes and Anton
Jurgens in the village of Oss. Also in the same locality was another family of
butter merchants-Simon and Henry Van den Bergh-who were to commence
production shortly afterward and who later were to join forces with the Jurgens
and eventually become the largest margarine manufacturers in the world
(Schwitzer, 1956).
Production followed in most countries in Europe-Austria (1873/4), Italy
(1874), Germany (1874), Norway (1876), and Denmark (1883), which was later
to become the largest per capita consumer of margarine. Mege applied for a
United States patent that was granted in 1874.
Many variations were developed in the following years and patents on new
formulations and processes were taken out. At the same time there was strong
opposition to the introduction of margarine into the market by the farming

Margarines and Spreads

257

community, and in some places antimargarine legislation was introduced.


Opposition from such quarters continued well into the twentieth century and is
not entirely unknown today.

10.3 Yellow-Fat Consumption


Despite the restrictions encountered in some countries, the consumption of
margarine has continued to grow, and in many cases has overtaken the consumption of butter. Consideration of available statistics may be complicated
since they do not always distinguish between the use of products for spreading,
baking, or frying. Nonetheless, the variation in consumption of spreads, especially before, during, and after the Second World War, is clearly marked. In
the United States, according to the National Association of Margarine
Manufacturers, the change in comparative levels of consumption of margarine
versus butter is significant. See Table 10.1.
Table 10.1

United States-Average consumption of butter and margarine (lb per


capita)

Year

Butter

Margarinet

1930
1935
1940
1945
1950
1955
1960
1965
1970
1975
1980
1985
1990
1991
1992
1993
1994

17.8
17.6
17.0
10.9
10.7
9.0
7.5
6.5
5.3
4.7
4.5
4.9
4.4
4.2
4.2
4.5
4.8

2.8
3.0
2.4
4.1
6.1
8.2
9.4
9.9
11.0
11.0
11.2
10.8
10.9
10.6
11.0
10.8
9.9

t From 1975; includes spread products.


Source: USDA Economic Research Service.

258

Food Emulsifiers and Their Applications

A similar development has been seen in Europe (Table 10.2), where in recent years (and also forecast to continue) the trend is for a reduction in yellowfat consumption generally, with small reductions in both butter and margarine
in the order of 4 to 5% and an increase in spreads of 17 to 19% by 1997.
Table 10.2 Europe-Average consumption of butter, margarine, and spreads
(kg per capita)

1987
Country
Belgium/
Luxembourg
Denmark
France
Germany
Greece
Ireland
Italy
Netherlands
Portugal
Spain

UK
Total EC

1992 (Est.)

Butter

Margarine

Spreads

Butter

Margarine Spreads

8.3
10.2
8.8
8.3
0.9
5.8
2.3
4.1
0.8
0.5
4.9
5.1

12.8
16.9
3.9
7.7
1.4
4.5
1.2
13.7
3.4
1.6
7.5
5.4

3.2
3.5
0.4
0.3
0.1
3.4

7.4
9.0
8.2
6.7
1.0
3.1
1.9
3.4
1.0
0.5
3.9
4.5

11.7
14.4
3.4
8.0
1.4
4.2
1.3
11.9
3.0

3.3
0.1
0.8
0.6
0.7

2.2

6.1
5.1

5.1
6.2
0.9
0.9
0.2
6.8
4.0
0.3
1.6
2.4
1.4

Source: Food Source '92 (1992), Frost & Sullivan, New York.

The reason for the change in consumption patterns in recent years-which


is expected to continue into the future-is principally related to concerns over
health following recommendations to reduce overall fat consumption and especially saturated fats from many authorities, for example, the National Academy
of Sciences in the United States and the Committee on Medical Aspects of
Food Policy (COMA) in the UK. Reduction in the consumption may also be
due to factors such as the decline in bread consumption.
There have also been significant structural changes within the markets, as
will be discussed later.

10.4

Definitions and Descriptions

The name and basic composition of margarine, or oleomargarine, has been


more or less established since Mege's original formulation-that is, it should

Margarines and Spreads

259

be similar to butter. However, with the new developments during more recent
years involving different components and varying combinations of vegetable
oils, animal fats, and milk fat, a range of new descriptions have come into being such as low-fat spreads, low-calorie spreads, and yellow-fat spreads. In
1993, Moran listed the varieties then available, as shown in Table 10.3.
Table 10.3

Some current spreads


Approx. fat
content(%)

High-fat spreads

Low-fat spreads

Butter
Margarine: Packet
Soft
Polyunsaturated fatty acids
(PUFA) spreads
Vegetable/butter-fatblended spreads
Vegetable fat spreads

80
80
80
80
80
70
60

40
Vegetable/butter-fatblended spreads
Butter-fat spreads
Very low-fat spreads
Water-continuous spreads

70

40
40
20-30
15
9

5
Source: Moran (1993).

In Europe, the adoption of Council Regulation (EC) No. 2991/94 of 5


December 1994 brought all products, including butter, under the one description, "Spreadable Fats" and described them as "products in the form of a solid,
malleable emulsion-with a fat content of at least 10% but less than 90% by
weight. The fat content, but excluding salt, must be at least two-thirds of the dry
matter." These terms are further qualified as "products which will remain solid
at a temperature of 20C and which are suitable for use as spreads."
The definitions and sales descriptions are detailed in Table 10.4.
Clause 2 of Article 3 of this regulation states that the sales descriptions
"minarine" and "halverine" may be used in place of "half-fat margarine."

10

l. Butter

A. Milk fats
Products derived exclusively from milk
and/or certain milk products for which fat
is the essential constituent of value

l. Blend

C. Fats composed of plant and/or


animal products
Products derived from solid and/or liquid
vegetable and/or animal fats suitable for
human consumption, with a milk-fat
content of between 10 and 80%

Source: Official Journal of the European Communities, 1316, Vol. 37,9/12/1994.

2. Three-quarter-fat blend
3. Half-fat blend
4. Blended spread xo/o
but not more than

l. Margarine
2. Three-quarter-fat
margarine
3. Half-fat margarine
4. Fat spreads xo/o

B. Fats
Products derived from solid and/or liquid
vegetable and/or animal fats suitable
for human consumption, with a milk-fat
content more than 3% of the fat content

2. Three-quarter-fat butter
3. Half-fat butter
4. Dairy spread xo/o

Sales description

Definitions and descriptions for spreadable fats

Fat-group definitions

Table 10.4

Not less than 39%; more than 41%


Products with fat content:
Less than 39%
More than 41 %; less than 60%
More than 62%; less than 80%
Product from a mixture of vegetable and/or animal fats
with fat content of not less than 80% but not more than 90%
Not less than 60%; not more than 62%
Not less than 30%; more than 41%
Products with fat content:
Less than 39%
More than 41 %; less than 60%
More than 62%; less than 80%

Milk-fat content not less than 80% but less than 90%.
Max. water 16%; max. dry nonfat milk materials 2%
Milk-fat content not less than 60% but not more than 62%
Milk-fat content not less than 39% but not more than 41%
Products with milk-fat content:
Less than 39%
More than 41 %; less than 60%
More than 62%; less than 80%
Vegetable and/or animal fats not less than 80%; less than 90%
Not less than 60%; less than 62%

Product categories
(additional description with
indication of% fat content by weight)

Margarines and Spreads

261

Whether the listed descriptions will gain greater prominence other than as
the required secondary descriptions is doubtful since consumers, presumably,
will prefer to favor the brand names that have been applied to the broad range
of products now available.
Fat spreads falling outside these standards, for instance those with fat contents below 10% and concentrated products with fat contents of 90% or more,
will not be classed as spreadable fats.
EC legislation does not specify vitamin fortification, which is left for decision at the national level. However, in the UK the requirement will remain as
previously specified in the Margarine Regulations (Sl 1867) 1967. That is,
each 100 g of margarine shall contain 800 to 1000 mg of vitamin A and 7.05 to
8.82 mg of vitamin D. This does not apply to other spreads, so that vitamin fortification in these cases is a matter for individual decision by the manufacturers and/or retailers.
Legislation in the United States and Canada is currently less detailed than
in Europe, but both require a minimum of 80% fat. In the United States, FDA
156.110 describes margarine (or oleomargarine) as "food in plastic form or
liquid emulsion" containing not less than 80% fat and describes a method for
fat determination. It specifies vitamin A fortification to be not less than 15,000
I.U. per pound but leaves vitamin D as an optional ingredient while stating not
less than 1500 I.U. per pound. While spreads are not specified in the legislation, low-fat spreads were stated in 1990 as the fastest growing category, with
spreads from 20 to 72% fat on the market (Borwanker and Buliga, 1989).
The National Association of Margarine Manufacturers of the United States
describes spreads as having fat contents ranging from 18 to 79%. Details of
market shares in the United States are shown in Table 10.5.

United States retail market share of


Table 10.5
margarine and spread products
Type

Stick margarine
Soft margarine
Liquid margarine
Low-fat vegetable oil spreads
Total retail market

1980

1990

65
19
1
15

44

100

100

l3
2

41

Source: National Association of Margarine Manufacturers.

262

Food Emulsifiers and Their Applications

Canadian Standard B.09.016 states that margarine shall be "a plastic or


fluid emulsion of water in fat, oil or fat and oil that are not derived from milk"
and shall contain not less than 80% fat and not less than 3300 I.U. vitamin A
and 530 I.U. of vitamin D.
Calorie-reduced margarine is specified in Standard B.09.0l7 as containing not
less than 40% fat and having 50% of the calories normally present in margarine.
The conversion of International Units of vitamins A and D to microgram
equivalents is complicated in the case of vitamin A since it depends upon the
origin of the fat. Thus, to convert vitamin A from I. U. to f..Lg retinol in foods of
animal origin, the I.U. of retinol should be multiplied by 0.3. However, for
foods of plant origin, the I.U. of beta-carotene should be divided by 10. This
arises because l I.U. of vitamin A is equivalent to 0.3 f..Lg retinol or 0.6 f..Lg
beta-carotene. The conversion of the units for vitamin D is straightforward, i.e.,
li.U. vitamin D = 0.025f..Lg.

10.5 Structure and Raw Materials


Margarine is a water-in-oil (W/0) emulsion; that is to say, the water (the disperse
phase) is distributed as droplets within the oil (the continuous phase). In margarine, the levels of each largely simulate that found in butter and are regulated
in most places by legislation, viz., minimum 80% fat, maximum 16% water, the
remainder consisting of salts, proteins, emulsifiers, vitamins, colors, and flavors.
Whereas production initially was based upon the use of fractionated animal
fats such as tallow and lard, these were later superseded by vegetable oils and,
for a period, marine oils, both of which offer a much greater flexibility in physical characteristics and economy.
Being the main and most expensive ingredient, the fat blend is also the
most important factor in the formulation of margarine. As oil prices fluctuate, it
is essential to have a fat blend providing the desired quality at minimum cost.
Thus, it is vital to be able to utilize alternative formulations based on prevailing price conditions while still maintaining quality. Therefore, a full understanding of the physical characteristics of the individual oils composing the
blend is critical. In this respect, factors of importance for the fat phase include
the solid/liquid fat ratio, crystallization rate, and melting properties.
In the early days of margarine production, large quantities of milk and water in almost equal proportions were used for emulsification with the fat, with
much of the excess water being pressed out during kneading to arrive at a final

Margarines and Spreads

263

moisture content of 16% or lower. Nowadays, the aqueous phase is added at


optimum levels and usually consists of a simple solution of skim milk powder
or whey powder in water, with the addition of salt and the pH adjusted to 5.5 to
6.0 by using, for instance, citric acid.

10.5.1

Oils and Fats

Oils and fats, otherwise known as triglycerides, are esters of glycerol and fatty
acids having a basic formula

where R 1, R2 , and R3 are the fatty acid residues, which may be the same but,
invariably, are different according to the source of the fat. Examples of the
variations in fatty acid compositions of different oils and fats are shown in
Table. 10.6.
Modification of the fat by, for instance, hydrogenation, whereby some of the
unsaturated links (double bonds) become saturated (for instance oleic C18:1 to
stearic CIS) offer the opportunity of considerable variation in the melting
points of fats, especially the liquid vegetable oils. The exact physical character
of a fat depends upon that of the constituent fatty acids that can vary widely in
melting point, as illustrated in Table 10.7.

10.6

Fat Crystallization

Variations in the crystallization properties of fats and differences between


batches of a similar origin may cause problems in production even though processing techniques are fully controlled and highly automated (Madsen, 1983).
The crystallization rates of some individual and blended fats are shown in
Table 10.8
Oils and fats are polymorphic and can crystallize in more than one form,
which can differ in terms of melting point, density, heat of fusion, and rate of
crystallization. The three crystal forms are commonly known as alpha (a), beta
prime (W), and beta (~), with the a form having the lowest values, the Winter-

""'

0.2
1.6
tr
25-30
2.5-3.5
15-25
35-45
7-12
0.5-1
1-4
59-70

0.2
3-4
0.5-1
26-28
2-3
23-27
30-35
1-1.5
0-1

45-57

4-8

Lard

9.2-13.9
tr
2.2-4.4
36.6-65.3
15.6-40.7
tr
2-8
84-105

7.7-9.7
2.3-3.2
5.4-7.4
1.3-2.1
tr
tr
7.5-10

tr

13-17

Groundnut

45.9-50.3
16.8-19.0

Coconut

tiodine value is a measure of the degree of unsaturation of the fat.

C-10 & lower;


capric, etc.
C12lauric
C14 myristic
C14:1 myristoleic
C16 palmitic
C16:1 palmitoleic
C18 stearic
C18:1 oleic
C18:2 linoleic
C18:3 linolenic
C20 & higher
Iodine valuet

Fatty acid

Beef
tallow

Table 10.6 Fatty acid compositions of oils and fats (%m/m)

tr
0.8-1.3
tr
43.1-46.3
tr
4.0-5.5
36.6-65.3
9.4-11.9
0.1-0.4
[xx]1
50-54

Palm

1.9-3.0
11.9-18.5
1.4-3.3
tr
[xx]1
16-19

7.2-10.0

43.6-51.4
15.3 17.2

Palm
kernel

3.4-6
0.2-0.6
l.l-2.5
52-65.7
16.9-24.8
6.5-14.1
1-6
109-126

tr

Lowerucic
rapeseed

9.9-12.2
tr
3.6-5.4
17.7-25.5
50.5-56.8
55-95
[xx]l.5
125-136

tr
tr

Soya

5.6-7.4
tr
3.0-6.3
14.0-34.0
55.5-73.9
tr
[xx]l.5
85

tr

Sunflower

Margarines and Spreads

265

Table 10.7 The melting points of fatty acids


Fatty acid

No. of double bonds

C12lauric
C14 myristic
C 14: 1 myristoleic
C16 palmitic
C16:1 palmitoleic
C18 stearic
C18:1 oleic
C18:2 linoleic
C18:3linolenic
C20 arachidic

M.P. eC)
44.2
54.4
Liquid
62.9
Liquid
69.6
16.2
Liquid
Liquid
74.4

1
1
1
2

Source: Andersen and Williams (1954).

Table 10.8

The crystallization rates of fats

Fat ( 0 C)

Time (min)

Temp.

Coconut
Coconut/palm 1:1
Coconut/palm, interesterified
Palm-hardened
Palm-hardened/palm 2:1
Lard
Lard/palm 1:1
Palm
Sheafat

3
4
5
5
8
14
15
27
45

20
15
18
17
13
lO
10
lO
10

Source: Madsen (1983).

mediate, and the most stable ~ form the highest values. Transition from one form
to another is from

W~ ~ and is not reversible without remelting the fat.

The polymorphism of crystalline fats may cause problems with the consistency
of margarine and spreads. During production, the fats initially crystallize in the a

form and normally will rapidly transform to the Wform. This is the desirable form
for spread production since the small needle-shaped

Wcrystals (about l!lm long)


Wform to the much

impart good plasticity. Should the crystals transform from


larger

p form (> 20 Jlm) they will give the spread a grainy consistency known as

"sandiness," as can be seen in Figure 10.1 (Madsen and Als, 1968).

266

Food Emulsifiers and Their Applications

(a)

(b)

Figure 10.1 Fat crystals in (a) normal margarine with good consistency and (b) a margarine with large ~crystals causing a "sandy" texture. (Magnification 200x.) (Courtesy

of Danisco Ingredients, Denmark.)

Concurrently, the specific area of the crystal surface will decrease, allowing
the liquid oil to penetrate to the surface of the margarine, which may then lead
to oiling out, especially if the margarine comes under pressure. Some vegetable oils such as partially hardened sunflower or low erucic rapeseed are
particularly prone to form ~ crystals and thus can cause sandiness in spreads.
In this case, sorbitan tristearate at 0.3% of the fat has been found to inhibit the

transition from the Wto the ~ form

The crystal-modifying effect of a range of emulsifiers-sorbitan esters,


ethoxylated sorbitan esters, ethoxylated fatty alcohols, citric acid esters of
monoglycerides (Citrem), diacetyl tartaric acid esters of monoglycerides
(DATEM), sucrose monostearate, sodium stearoyl lactylate, and polyglycerol
esters- has been investigated on the polymorphism of tristearin (glyceryl tristearate) (Garti et al., 1982).
In these trials it was found that sorbitan monostearate and Citrem (Cl6/Cl8
fatty acids) were the most effective in preventing the recrystallisation (from a
to ~ form) of tristearin.
It should be noted that when used in emulsions, surface-active materials will

Margarines and Spreads

26 7

be adsorbed at the oil/water interface and that only lipophilic emulsifiers with
high solubility in the oil phase can perform as crystal inhibitors in emulsions.
Varying the level of saturated fat affects the degree of crystallinity of the oil
phase (Borwanker and Buliga, 1989). Spreads with 40% fat were made using
oil phases of varying crystallinity by blending either liquid soyabean oil or
stick margarine oil with soft margarine oil. The textures of the spreads made
with these blends were visibly very different, increasing in softness in the
same direction as the oil itself. As a consequence of increased firmness, the
stability of the resulting spread was expected to increase with increasing crystalinity in the oil phase. A centrifugation procedure was used to measure the
stability of the emulsions (Figure 10.2). The higher amount of water and/or oil
(especially water) released corresponds to a less stable emulsion.

10.7

Emulsifiers

The main function of emulsifiers in processed foods is to reduce the interfacial


tension between the phases of an emulsion-usually oil and water. In such
two-phase systems, one phase is dispersed as large droplets within the other.

60
"0

+-'Cil

Ul

Cll co
u Cll
Cll Cll

....

.s-.:
>.0
..~I...

==o
..c.._
co Cll
+-'+J
CJ)co

3:

""' '"--

50
40
30

-a...___

20

O Oil

"-~

10

O Water

0
4.5

6.3

8.6

10.4

13.0

Crystallinity (enthalpy, J/g)

50 Soft
50 liquid

75

25

Soft

75

25

50 Soft
50 Stick

Figure 10.2 Effect of the degree of crystallinity on the emulsion stability of 40% fat
spreads as measured by ultracentrifugation. (From Borovanker and Buliga, 1989.)

268

Food Emulsifiers and Their Applications

They are either oil-in-water (0/W) emulsions, where the continuous phase is
water (such as in milk or ice cream) or water-in-oil (W/0), where the continuous phase is oil (as in butter and margarine).
The stability of two-liquid phase emulsions is kinetic stability, i.e., the system
is not thermodynamically stable (Friberg et al., 1990). Thermodynamically stable emulsions would spontaneously reform following separation by, for instance,
centrifugation, while experience shows that an emulsion that has separated remains so unless mixed by some external action. In reality, the two separated
phases are in the most stable state to which all emulsions will tend. Thus, a stable emulsion is one in which this inevitable trend has been retarded so that it is
not noticeable during the normal life of the product--even if it be several years.
Margarine is a water-in-oil emulsion in principle only because, in reality, it is
a dispersion of water droplets in a semisolid fat phase, containing fat crystals
and liquid oil (Krog et al., 1983). Preparation of the emulsion requires considerable energy to reduce the droplet size of the disperse phase, thereby creating an
increase in the surface area between the two immiscible phases.
The initial water-in-oil emulsion is prepared in mixing tanks with vertical or
horizontal stirrers to ensure emulsification but without incorporating too much air.
It is rather coarse in water-droplet size and fairly unstable if not kept agitated.
In modern continuous production, the emulsion exists for only a brief period before passing into the chilling unit where final emulsification and crystallization of the fat phase take place. Thus, the emulsion need not be very
stable against coalescence as the water droplets become fixed within the semisolid phase. However, the droplet size is important when considering flavor release and microbiological spoilage. Thus, emulsifiers are used to lower the
interfacial tension between the oil and water phases, which will generally result in a smaller water-droplet size. Distribution of the water droplets in the
range 2 to 4 11m will give better stability against deterioration such as mold
growth. However, it is still desirable for some of the water droplets to be larger
in size-10 to 20 11m-to give a better flavor release in the mouth.
For these purposes, therefore, lipophilic emulsifiers such as mono-/diglycerides of long-chain fatty acids (C16-C18) are used at levels of 0.1 to 0.3%,
often in combination with 0.05 to 0.1% refined soya lecithin.
Comparisons of the water-droplet distribution in margarine emulsion and in
finished margarine are shown in Figure 10.3. Droplet size in the emulsion is
further reduced during the cooling and kneading processes in the tube chiller.
The water droplets in finished margarine are stabilized by adsorbed fat

Margarines and Spreads

(a)

269

(b)

Figure 10.3 Water-droplet size distribution in (a) margarine emulsion and (b) the finished margarine. (Magnification 200x.) (Courtesy of Danisco Ingredients, Denmark. )

crystals, as seen in Figure 10.4 showing the microstructure of margarine by


freeze-fracture transmission electron microscopy. It is clearly seen that the water droplets are covered by fat crystals, oriented flatly along their surface.
Margarine is frequently used for frying when it is especially important that
it does not spatter. Spattering is caused when the margarine melts in the frying
pan, the emulsion breaks, and the coalesced water droplets, due to gravity,
form a film of water covered with molten fat on the frying pan. When the temperature reaches boiling point, the increase in vapor pressure will cause spattering-sometimes explosive-of the water phase.
It is desirable, therefore, that the margarine allow gradual evaporation of
the water from the small water droplets and the formation of a fine, golden
brown sediment that does not adhere to the frying pan. Both formulation and
processing play important roles in reducing the tendency to spatter. The presence of salt and milk are desirable as is a high pH, up to say 6, while sugars
and starches will increase the tendency to spatter. The selection and addition
of the emulsifier is of considerable importance in producing a margarine with
good frying properties.
While a combination of mono-/diglycerides and lecithin will have only a

270

Food Emulsifiers and Their Applications

Figure 10.4 Freeze fracture electron micrograph showing the microstructure of margarine. Water droplets (w) are covered with fat crystals. (Bar: l!lm.) (Courtesy of Dr. W.
Buchheim, Keil.)
limited effect in reducing spattering in low-salt margarine, there are other
emulsifiers that, either alone or with lecithin, will help prevent coalescence of
water droplets during frying. In this respect, citric acid esters of mono- and
diglycerides (Citrem), polyglycerol esters, and thermally oxidized soybean oil
interacted with mono- and diglycerides used at 0.3 to 0.4%, together with soya
lecithin can be very effective.

10.8 Processing
Typical modern processing involves the separate processing of the oil and water phases. The oils and fats are kept in storage tanks at temperatures just adequate to maintain them at the required fluidity. Preferably, they should be
bottom fed to avoid splashing and aeration. It is essential that the plant, i.e.,
tanks, pipes, and pumps, be entirely free from copper or copper alloys to avoid
the high risk of oxidation.
The final oil blend together with oil-soluble components such as emulsifiers, colors, flavors, and vitamins, is prepared separately from the water phase

Margarines and Spreads

271

that may contain milk components such as whey powder, skim milk powder, or
sodium caseinate and also salt, gelatin, or thickener, water-phase flavors where
appropriate, and preservatives such as potassium sorbate for low-fat products.
Modern methods involve the feeding of the phases by computer-controlled
load cells, which ensures fast throughput and constant composition.
A second emulsion tank is used for maintaining quantities of finished emulsion for feeding the cooling and kneading equipment. Cooling and kneading
can be carried out by either of two different methods: (i) tube chiller or (ii)
chilling drum-complector.
In the former, cooling and kneading take place in a closed system and in a
single process. In the latter, the cooling and kneading are carried out separately-cooling on the chilling drum and kneading in the complector. The advantage of the chilling drum-complector method is that it allows the product to
rest between cooling and kneading, which is important with formulas based on
slower crystallizing fats, for instance, puff pastry margarine. By comparison,
the tube chiller is relatively more compact considering throughput, is easy to
operate, and reduces the possibility of spoilage.
Most types of margarine can be made very satisfactorily using the tube
chiller method, although in some cases extra working units (pinning machines)
may prove advantageous. However, the chilling drum-complector has been the
preferred method for puff pastry margarine, although tube chillers are now
widely used, and is perfectly satisfactory for normal table margarine and for
cake margarine. The suitability of both methods is summarized in Table 10.9.
Table 10.9

Comparison of processing methods


Chilling
drum-

Type

Tube chiller

complector

Block/stick margarine
Soft table margarine
Low-fat spread
Cake and creaming margarine
Puff pastry margarine
Shortening
W/0 emulsion with high water
content

XXX

XXX

XXX

XXX

XXX

XXX

XX

XXX

XXX

XXX

XXX

= Excellent;

XX

= acceptable; X = poor.

Source: Madsen (TPlOl).

272

Food Emulsifiers and Their Applications

The comparative merits of the two methods were investigated using four different fat combinations (see Table 10.10) in puff pastry margarines that were then
evaluated by various parameters including a "finger" judgement (see Table
10.11) of plasticity after one week (Madsen, TPlOl).
Table 10.10

Dilatation, iodine value, and melting points of fat blends used for
comparison of puff pastry margarines
Solid fat index
Fat blend

so

10

15

20

25

Iodine
value

l.
2.
3.
4.

37
40
43
37

37
40
42
35

34
36
37
31

27
31
29
25

22
24
23
19

67
7l
66
74

l8Ct animal
18Ct vegetable
12Ct animal
soct animal

M.P.

eq

37
37
41
37

tThe figures indicate storage and rolling temperature of the puff pastry margarine.
Madsen (fPlOl).

Source:

Table 10.11

Plasticity judgement by finger method

Fat blend

Method

Plasticity judgment

Score

l. l8C animal

Chilling drum
Tube chiller

5
0

2. 18C vegetable

Chilling drum

Fine plasticity
Lumpy/gritty, a little firm,
becoming slack and very
greasy when worked
Good plasticity, tending to
become greasy
Gritty, firm, becoming slack
and greasy when worked
Fine plasticity

Tube chiller
3. 12C animal

Chilling
Drum
Tube chiller

4. soc animal

Source:

Madsen, TPlOl.

Chilling drum
Tube chiller

Short and gritty, very,


firm, becoming soft and
greasy when worked
Fine plasticity
Short and gritty, very firm,
becoming soft and greasy
when worked

4
1
5
0

5
0

Margarines and Spreads

273

Similar results were found when the same products were judged after 3
weeks' storage.
This rather critical evaluation did not fully take into account the differences that might result from the use of varying techniques possible with tube
chillers, which was investigated separately.

Table 10.12 Plasticity judgment by means of the finger method in puff pastry
margarines made on tube chiller with varying production techniques
Production
technique

Fat blend

l. Normal

Animal
Vegetable

2. Intermediate
crystallizer

Animal
Vegetable

3. Pinning machine

Animal
Vegetable

4. Resting tube +
pinning machine
5. Low capacity

Animal
Vegetable
Animal
Vegetable

Plasticity judgment
after I week at l8C

Points

Lumpy and gritty, a little firm but


becomes slack and greasy when worked
Gritty, firm but becomes slack and
greasy when worked
A little lumpy and soft, very greasy
when worked
A little firm but becomes soft and
greasy when worked
A little lumpy and fairly soft; very
greasy when worked
Becomes easily soft and greasy when
worked
Soft and greasy

Too soft and very greasy


Fine homogeneity but too soft, fairly
good plasticity
Fine homogeneity, a little too soft,
fairly good plasticity

l
3

l
1

2
l

2
0

Source: Madsen, TPlOl.

The important issue relates to the postcrystallization of the fats, which was
further investigated (Madsen, 1981 ). A good plastic puff pastry margarine can
be bent without breaking and the plasticity evaluated by repeated handworking to check stability, firmness, and greasiness (Figure 10.5).
In the case of cake margarine, the most important points are that the fat
blend is soft and easy to incorporate into the batter and has good creaming
properties. This, therefore, suggests the use of lauric fats (coconut, palm kernel) that crystallize quickly and, thereby, facilitate creaming.

274

Food Emulsifiers and Their Applications

Figure 10.5 Evaluation the plasticity of puff pastry margarine


by hand. (Courtesy of Danisco Ingredients, Denmark.)

10.9 Reduced- and Low-Fat Spreads


As has been previously highlighted, low-fat spreads have been the only growth
area in a gradually declining market. They can have fat contents from lO to
79% and, except in the case of very low-fat spreads, are W/0 emulsions.
However, with low-fat products, it is necessary to strike a balance between stability and mouthfeel, which can be affected both by the composition and the
method of processing.
Margarine is essentially a stable product and the appropriate fat blend and the
combination of milk proteins, lecithins, and/or saturated mono- and diglycerides
used in production ensure both stability and the desired eating characteristics
(Flack, 1992). However, in reduced- and low-fat spreads in which the dispersed
phase (aqueous) can exceed the phase volume of the continuous (fat) phase, problems can arise with stability, meltdown, and flavor release (Moran, 1993).
While milk proteins are in high-fat products to improve mouthfeel and flavor, they also act as hydrophilic 0/W-emulsion stabilizers, and thus their use
in low-fat products, together with the considerable energy input needed, could
result in phase inversion.
These problems can be overcome by a combination of formulation and processing. Important factors in the formulation include the melting characteristics

275

Margarines and Spreads

of the fat blend, the type and level of emulsifier, and the addition of thickeners
such as gelatin, sodium alginate, pectin, and carrageenan to the aqueous phase.
Low levels of whey protein may be used to improve flavor release with the further
advantage that the pH of the aqueous phase can be lowered to improve keeping
properties because, unlike casein, whey proteins do not precipitate at low pH.
The speed of processing, i.e., the rate of throughput and the emulsion temperature, are also important factors in the stability of the spread.
Examples of the effect of various types of emulsifiers on the stability of lowfat spreads with fat contents of 40% and 20% are shown in Tables 10.13 and
10.14, respectively.

Table10.13

Effect of types and blends of emulsifiers on the stability of a 40% fat

spread
Water separation % after

Levels
of use

Types of emulsifier

5 min

10 min 15 min 20 min

0.6%

Distilled monoglyceride from

1.6

2.6

9.5

15.8

l.O

2.6

6.3

0.6%
plus
0.2%

0.4%
plus
0.2%

vegetable oil. IV approx. 80


Distilled monoglyceride from
vegetable oil. IV approx. 80
Polyglycerol ester of fatty
acids. IV approx. 80
Distilled monoglyceride from
vegetable oil. IV approx. 80
Polyglycerol ester of interesterified
ricinoleic acid. IV approx. 85

IV = Iodine value.
Formulation of spread:
Water phase

56.4% water
0.5% whey powder

pH 4.5 1.5% gelatin

1.5% salt
0.1% potassium sorbate
Fat phase

39.2-39.4% fat blend


0.{H).8% emulsifier (as above)
4 ppm beta-carotene

Source: Madsen (1989).

276

Food Emulsifiers and Their Applications

Table 10.14 Effects of types and blends of emulsifiers on the stability of a 20% fat spread
Levels and types of emulsifiers

Results

0.8 % distilled monoglyceride from


sunflower oil. IV approx. lOS
O.S % distilled monoglyceride from
sunflower oil. IV approx. lOS
plus
O.S% polyglycerol ester of
interesterified ricinoleic acid. IV
approx 8S

Emulsion split into separate


phases in tube chiller
Produced a fine, stable spread
with good spreadability and
mouthfeel

IV = Iodine value.

Formulation of spread:
Water phase

69.9% water
4.0% skim milk powder

pH6.8

3.0% gelatin

l.S% sodium alginate


1.5% salt

0.1% potassium sorbate


Fat phase

19.0-19.2% fat blend


0.8--1.0% emulsifier (as above)
8 ppm beta-carotene

Source:

Madsen (1989).

The trial batches in both cases were made on a three-tube Perfector pilot
plant using a fat blend composed of
60 parts soya oil
30 parts partially hydrogenated soya oil
lO parts coconut oil
Both examples indicate the potential instability of these emulsions and the
variation in the stabilizing effects of different emulsifier blends even when used
at low dosage levels. These results suggest that emulsifier structure at the interface plays a critical role in retarding water-droplet flocculation and coalescence.
While the evaluation of spreads by measuring water separation provides
quantifiable results, a quicker and more practical evaluation of the stability of

Margarines and Spreads

277

spreads can be made by spreading a sample with a knife on cardboard, as


shown in Figure 10.6, where the desired qualities of good spreadability and
low moisture loss are shown in the right-hand photograph. A rheological measurement to obtain quantitative data could also be carried out.

It is rather more difficult to produce low-fat butter spread from butter oil
due to the relative hardness of the fat at low temperatures. Nonetheless, spread
with a butter fat content of about 40% has been produced using 5% sodium caseinate plus 2% sodium alginate in the water phase and 0.5% distilled monoglyceride, IV approximately 55, in the fat phase.

Figure 10.6 Evaluating margarine by spreading on cardboard. (Courtesy of Danisco


Ingredients, Denmark.)

However, in this case it is not possible to lower the pH of the aqueous phase
without precipitating the caseinate, which will reduce its emulsifying properties, and therefore, the keeping properties of the spread will be limited.
However, a satisfactory low-fat butter spread can be produced from dairy
cream, using a distilled monoglyceride with high iodine value (80 to 105) in
the cream and a thickener such as sodium alginate. In this case, phase inversion from 0/W to W/0 can be achieved in the tube chiller, using normal or
slightly reduced cooling and operating at 40 to 50% of the usual capacity. To
obtain a satisfactory working effect, a high rotor speed in the tube chiller cooling cylinder would be preferable (Madsen, 1989).
Factors that may affect the efficiency of phase inversion in low-fat emulsions are listed in Table 10.15.

278

Food Emulsifiers and Their Applications

Table 10.15

Factors promoting the inversion of oil-in-water emulsions

l. Increased rotor speed


2. Controlled temperature (maintenance of optimum fat solids level)
3. Use oflow-HLB emulsifiers
4. Increased oil-droplet size entering unit
Source: Moran (1993).

Most of these factors concern the increase in rate of collision of oil droplets
upon which the rate of coalescence is dependent (Moran, 1993). The emulsifier(s) likely provide steric hindrance to keep droplets separated.

10.10 Oil-in-Water Spreads


According to Moran (1993), the advantages of 0/W spreads over the more conventional W/0 spreads may be attributed to

Product structure not dependent on the type of fat used

Any level of fat can be used from 1% to more than 50%


High levels of protein are possible

Processing is easier and cheaper

Flavor release is quicker on the palate

Against this, however, one major drawback is that unless products are prepared at comparatively low pH, then ultra-high-temperature processing and
possibly an aseptic filling procedure must be followed if shelf lives comparable
to conventional spreads are required.
Oil-in-water spreads remain a relatively unexplored area of the spreadable
fats market, possibly due to the problem of microbiological deterioration, but
may offer a potential for future expansion in markets where adequate levels of
preservatives are permitted. In contrast to W/0 emulsions, these systems
would utilize high-HLB emulsifiers to stabilize the water-continuous emulsion.

10.11 Liquid Margarine


Liquid margarine is used primarily for frying and is available particularly in
markets such as the United States, Germany, and Sweden, where butter or
margarine are normally used for frying (see Table 10.5).

Margarines and Spreads

279

In markets such as the UK that have traditionally used solid fats like lard or
cooking fat or, more recently, liquid vegetable oils, there is less interest in using liquid margarine.
A typical composition has a fat phase of about 82% based on soya or sunflower oil, in which emulsifiers perform two functions. First, an emulsifier such
as citric acid ester of monoglyceride (Citrem) at, say, 0.4% plus 0.2% soya
lecithin produces a stable water dispersion with limited spattering during frying; and second, a selected mono- and diglyceride blend prevents oiling out on
storage and thus gives a homogenous product with low viscosity. If the margarine is not intended for frying, the Citrem can be replaced with 0.2% distilled monoglyceride. The water phase may contain proportions of skimmed
milk powder and salt as well as a preservative such as potassium sorbate.
Flavoring may be added to both phases.
The aqueous phase should be adjusted to 4.5 to 6.5 pH and pasteurized at
80C. The fat phase should be tempered to about 60C and the emulsifiers
melted into a small amount of the liquid oil to a temperature of 65 to 70C,
which is then stirred into the main part of the fat phase. The water phase is
added to the fat phase under continuous agitation before cooling through the
tube chiller with an outlet temperature of approximately 5 to 7C.
The emulsion should be rested for 2 hours to allow proper fat crystal formation and then stirred vigorously for 15 minutes before topping off for packaging.

10.12

Summary

Standard margarines containing 80% fat are, under normal conditions, relatively stable, requiring minimal quantities of lecithin and/or mono- and diglycerides over and above the quantities of milk proteins usually present.
However, where the margarine is required for special purposes, i.e., cakemaking or frying and particularly in products with reduced fat levels, emulsifiers specific to the unique functional requirements of the application should
be selected (see Chapter 8 for applications of emulsifiers in baking products).

References
Andersen, A.J.C., Williams, P.N. (1954). Margarine, 2d ed., Pergamon, Oxford.
Borwanker, R.P., Buliga, G.S (1989). Emulsion properties of margarines and low fat
spreads, in Proc. Food Emulsions and Foams-Theory and Practice (eds. P.G. Wan
eta!.), San Francisco, pp. 44-52.

280

Food Emulsifiers and Their Applications

Flack, E. (1992). The role of emulsifiers in reduced fat and fat free foods, in Food
Technology International Europe (ed. A. Turner), Sterling Publications, London, pp.
179-181.
Friberg, S.E. et al. (1990). Emulsion stability, in Food Emulsions (eds. K. Larrson and
S.E. Friberg), Marcel Dekker, New York, pp. 1-40.
Garti, N., et al. (1982).]. Am. Oil Chem. Soc., 59, 181.
Krog, N., et al. (1988). Applications in the food industry, in Encyclopaedia of Emulsion
Technology, Vol. 2: Applications of Emulsions (ed. P. Becher), Marcel Dekker, New
York, p. 321.
Madsen, J. Puff Pastry Margarine: A Comparison of the Chilling Drum and Tube Chiller
Methods of Production, TP101, Grinds ted, Denmark.
- - - (1981). Post-Crystallisation in Puff Pastry Margarine, at 11th Scandinavian
Symposium on Lipids, Bergen, Norway.
- - - (1983). Product Formulation and Processing of Margarine and Yellow Fat
Spreads, at Margarine and Yellow Fat Seminar, Coventry, England.
- - - (1989). Low Calorie Spread and Melange Production in Europe, at World
Conference on Edible Oils and Fats Processing, Maastricht, Holland.
- - - , Als, G. (1968). Sandiness in Table Margarine and the Influence of Various
Blends of Triglycerides and Emulsifiers Thereon, at /Xth Int. Soc. of Fat Research
Congress, Rotterdam.
Moran, D.P.J. (1993). Reduced calorie spreads, in Porim Technology, No. 15., Feb '93,
Palm Oil Research Institute of Malaysia.
Schwitzer, M.K. (1956). Margarine and Other Food Fats, Leonard Hill, London.

ELEVEN

Emulsifier Trends
for the Future
Gerard L. Hasenhuettl

As in many other industries, forecasting the future is usually done by examining the trends of the past and extrapolating the changes for several years into
the future. While this works well for continuous functions, discontinuous
events often produce dramatic changes in market structures. Forecasting technology is essentially an observation of technical events, extrapolating the
trends, and trying to estimate which technologies are about to jump to the next
S curve. Food technology abhors radical change. Development of new crops is
contingent on being able to diffuse the crop into the massive agronomic supply
chain. Issues such as cross-pollination and identity preservation may pose significant problems. When a new product is brought to market, it needs to be
thoroughly tested or examined for its safety. Programs like HACCP are aimed
at institutionalizing safety into the industry. Even if the government is assured
of a product's safety, consumer acceptance is by no means assured.
Controversies around genetically engineered and irradiated foods are testimony to this principle.
As we stated in Chapter l, development of totally new emulsifiers is unlikely because of the time and cost involved with regulatory approval.
However, this by no means sounds a death rattle for innovations in the field of
281

282

Food Emulsifiers and Their Applications

food emulsifiers. Some social, demographic, and technical developments may


cause discontinuities in the underlying science involving food emulsions. A
few of these trends will be examined.

11.1

Globalization in the Food Industry

The food industry has traditionally been multidomestic. A number of factors


such as local taste, perishability, and low value-to-volume ratios in shipping
were responsible. However, counter trends such as increased income in developing nations, more international communication and travel, and increased consolidation in the processed food industry are exerting globalization pressures.
Development of food products for the global market will change some of the
rules for higher value products. Emulsions will require more microbiological
and physical stability to withstand shipping long distances and the absence of
refrigerated storage in some countries. Some lessons may be learned from the
pharmaceutical and cosmetic industries that are further advanced in the application of emulsifiers to produce long shelf lives.
One example of a new global product might be a margarine that is stable
without refrigeration. In addition to a higher melting fat, a more robust emulsifier system will be required because coalescence of water droplets could cause
separation into a phase that would be vulnerable to microbial growth.
As global population continues to explode, food emulsifiers will also need
to adapt to new lower cost sources of protein, carbohydrate, and fat emulsions
formed at lower temperature and mechanical shear can also contribute to energy savmgs.

11.2 Nutritionally Driven Changes in Foods


Nutritional studies in the area of diet and health or the counterpart, diet and
disease, are continually being carried out and reported, often with apparently
contradictory interpretations. Assessing trends in this area can be confusing
unless one stays with a few areas where there is general consensus.
Saturated fat has been long associated with its ability to raise serum cholesterol, notably LDL. The FDA's NLEA regulations recognized this consensus by
requiring labeling of saturated fat for all packaged foods. Food processors have
therefore been making a substantial effort toward substitution of solid fats with
liquid oils. This poses a formidable challenge in some products where solid fat is

Emulsifier Trends for the Future

283

part of the physical structural or primary functionality of food. For example, the
development of high liquid-oil margarines will require an emulsifier that can stabilize a W/0 emulsion where the continuous phase is liquid. Adjunct technologies, such as the development of molecular networks to thicken the oil phase can
make a significant contribution to providing structure to the processed food.
Solid fat is implicated in such functions as aeration of whipped toppings and formation of evenly dispersed air cells in cakes. Replacement with liquid oil places
more of a functional burden on the emulsifier system.
Health authorities have recommended that total dietary fat should not exceed 30% of calories. Some consensus is also building that some forms of cancer may correlate to total caloric consumption. Since fat represents 9 cal/gram,
cutting fat consumption will also reduce total calories (if the same weight of
food is consumed). If consumers respond to these recommendations, pressures
for availability of reduced-fat and fat-free foods will continue or even increase.
Elimination of fats from products normally containing high levels creates enormous challenges to maintain texture and flavor. Flack (1992) has speculated
that emulsifiers can play an important role in reclaiming some of these qualities. Structured emulsifier phases have been patented as the basis for formation of a pseudo-fat phase (Heertje, et al., 1994; Kleinherenhrink et al., 1995).
This trend toward replacement of fat functionality with emulsifiers will undoubtedly continue.

11.3 Trends toward Safer Emulsifiers


Interest in natural and minimally processed foods has waned and resurged
many times over the last half century. As previously mentioned, phospholipids
and proteins are the only "all natural" emulsifiers. The discovery and elimination of dioxane in polysorhates illustrates many of the concerns voiced about
synthetic food additives. Testing and surveillance of additives will continue
and more definitive assays will he developed to assure their safety.
Toxicological evaluations are also becoming more sophisticated and specialized. For example, Larsson (1994) has described a model that tests the effects of surfactants on gastric and intestinal lumina cells. Concerns about
penetration of single-chain food emulsifiers, such as lysophosphatidylcholine
and sodium stearoyllactylate, have been expressed.
Microbes have long been used in the food industry to produce cultured
products such as cheese and yogurt. Microbes have also been used to produce

284

Food Emulsifiers and Their Applications

very efficient surfactants for application such as enhanced oil recovery and oilspill control. A logical extension of these technologies would be to develop
food-grade organisms that produce natural surfactants. However, extensive
evaluation would still be essential to ensure the safety of these products when
consumed in processed foods.

11.4 Emulsifier Structure and Interactions


with Other Ingredients
Chapter 6 of this book has discussed the formation of mesophases of food
emulsifiers in aqueous environments. Although this property has been known
for a few decades, functional implications are just beginning to emerge
(Heertje et al., 1994; Kleinherenbrink et al., 1995). lsraelachvili (1985) has
described mesophase formation as a function of intermolecular forces. He proposes a model, shown in Figure 11.1, where a free energy minimum is
achieved by formation of a structure that balances attractive forces of hydrophobic alkyl chains and the repulsive forces between polar head groups.
The author has calculated a "critical packing parameter" from the effective
head group area, length of the alkyl chain, and volume of the cone defined by
the previous two factors. Table 11.1 lists the structures expected for ranges of
critical packing parameters. The significance of this technology is that once

Interfacial
(hydrophobic)
attraction

, . - - / H e a d-group (hydrophilic) repulsion

Volume v

-._ \

Area a 0

Figure 11.1

Determination of critical packing parameter {lsraelachvili, 1985).

Emulsifier Trends for the Future

Table 11.1

285

Amphiphilic packing of surfactants. (From lsraelachvili, 1985.)

Lipid type

Critical
packing
shape

Critical
packing
parameter (VIaJ.)

Structures
formed

Single-chain lipids
with large head
group areas
Single-chain lipids
with small head
group areas
Double-chain lipids
with large head
group areas
Double-chain lipids
with small head
group areas
Double-chain lipids
with small head
group areas

< l/3

Cone

Spherical micelles

l/3-l/2

Truncated cone

Cylindrical micelle
(hexagonal)

l/2-l

Truncated cone

Flexible bilayers,
vesicles

-l

Cylinder

Planar bilayers
(lamellar)

>l

Inverted truncated
cone

Inverted micelles

the functional role of a mesophase is defined, rational design of head and tail
groups can he accomplished and the emulsifier synthesized by techniques described in Chapter 2.
Emulsifiers may he viewed as one class of functional food ingredients. As
Chapters 4 and 5 have pointed out, these ingredients may interact with carbohydrates and proteins to produce a number of technical effects. The area of ingredient interactions is the subject of a recent book by Gaonkar (1995). New
experimental techniques such as scanning tunneling microscopy and interfacial rheology have the potential to provide greater understanding of these interactions. Emulsifiers can then be designed to form more selective interactive
structures that may result in improved or entirely new functionalities.

11.5

Enzymatic Synthesis of Food Emulsifiers

Production of emulsifiers involves contacting lipid and polar phases that are
mutually insoluble. To achieve homogeneity, temperatures as high as 500F
are applied and held until reaction is complete.

286

Food Emulsifiers and Their Applications

A consequence of this drastic processing is the production of undesirable


and often unidentified byproducts from side reactions. These byproducts can
impart metallic, soapy, or bitter off-flavors in finished food products. For example, high temperatures cause caramelization of sucrose esters. To avoid this
problem, solvents such as dimethyl sulfoxide and dimethyl formamide have
been used. However, removal of these solvents from the product poses another
serious problem.
An alternative approach is to catalyze esterification or interesterification reactions with enzymes such as lipase or esterase. Since these reactions are carried
out at much lower temperatures, formation of odors and off-flavors can be significantly reduced. This is particularly advantageous for unsaturated fatty acids that
can be oxidized by high temperatures and small quantities of air. Another potential advantage for enzyme synthesis would be the ability to produce high concentrations of monoesters without the need for short path distillation. To accomplish
this, the reaction could be carried out at a temperature just below the melting
point of the monoester. As the reaction proceeds and monoester selectively precipitates, the equilibrium will shift to produce more monoesters.
Examples of emulsifier synthesis are recent works of Vulfsen (1993) as well
as Arcos and Otero (1996). Shah and coworkers (1994) have studied lipase-

catalyzed reactions in monolayers and microemulsions. The major disadvantages of enzyme-catalyzed interesterification are high enzyme cost (relative to
sodium or calcium hydroxide catalysts), longer reaction times, and the tendency of enzymes to denature. However, the problems are surmountable and
the use of enzyme technology is expected to increase.

References
Arcos, J.A., Otero, C. (1996). ]. Am. Oil Chem. Soc., 73:673-82.
Flack, E. (1992). Food Techrwlogy International-Europe, 79-81.
Gaonkar, A. (ed.) (1995). Ingredient Interactions: Effects on Food Quality, Marcel
Dekker, New York.
Heertje, 1., et al. (1994). European Patent 0 558 523 Bl, July 13.
lsraelachvili, J.N. (1985). Intermolecular and Surface Forces: With Applications to
Colloidal and Biological Systems, Academic, New York, pp. 246--62.
Kleinherenbrink, F.A., et al. (1995). International Patent Application WO 95/35035,
December 28.
Larsson, K. (1994). Lipids-Molecular Organization. Physical Functions and Technical
Applications, The Oily Press, Ayr, Scotland, pp. 156--62.
Shah, D.O., et al. (1994). ]. Am. Oil Chem. Soc., 71:1405-9.
Vulfsen, E.N. (1993). Trends Food Sci. & Techno!., 4:209.

Index

Index

hydrocarbon chain packing, 151


thermodynamics, 150
albumin, 102
"all natural" ingredients, 5
amino acid, peptide chain, 97
amphiphilic packing of surlactants,

acetylated diglycerides, 33
acetylated monoglycerides, 33, 44,

60,253
preparation of, 33-36
acetylated monoglyceride citrate,

285

33-36

ampoules, flame-sealed, 153


amylopectin molecules, 68, 79, 80
amylose-complexing index, ACI, 252
amylose molecules, 68, 79
amylose leaches, 226
amylopectin, 226
anionic emulsifier:
sodium stearollactylate, 219
anionic surfactants, 221
AOAC, Association of Official
Analytical Chemists, 40, 56
method, 51
AOCS, 56
association structures, 154-155
associative adsorption, 167, 169

preparation of, 33-36


acid value/free fatty acid, 44
adsorbed proteins:
binding strength of, 101
interactions of surfactants, 101
influence of, 102
adsorbed surfactant molecules, 101
adsorption, emulsifier:
associative, 167
competitive, 167
layer, 167
surface, 167-169
aeration, bakery products, 214, 217
aggregation structures:
association structures, 151, 152

289

290

Index

B
baked products:
cream icings, 232
crumb-softening, 226
emulsifiers, 216
applications, 225
extruded snacks/cereals, 232
fat-free products, 232
dough conditioning, 225
monoglycerides in, 217, 218
production influences, 229
shortening, 215
yeast in, 217
Bancroft rule, 162, 166
birefringence, loss of, 78
bovine milk, 173
composition of, 174
bovine serum albumin 98 102 113
' '
'
'
126
bread dough, 85

Brewster angle microscope (BAM),


96
Brookfield viscosity measurements
'
241
butter, 1, 174, 202, 257, 259
butter and margarine consumption:
content of, 259, 260
in Europe, 258
market share of, 261
in the U.S., 257
butterscotch, 251

c
cake batters, 212, 230
calcium stearoyl-2-lactylate, 20, 57
calcium hydroxide, 17
capillary melting point, 53

carboxylic acid group reaction, 20


caramels, 251
casein, 122, 124, 126, 134, 170
micelles, 173
caseinate, 134
casson viscosities, yield values:
chocolate, 243, 244
cationic surfactant (CTAB), 199
cetyl-trimrthyl ammonium bromide
'
CTAB, 199
cheese, 174
manufacture, processed, 198
chemically leavened products, 229
cookies, crackers, 230
layer and snacks cakes, 229
spread ratios, 231
chewing gum, 250
lecithin, 250
mono- and diglycerides, 250
chocolate, 7
antibloom agents, 248--249
bloom, 245--248
coatings, viscosities, 244
compound coatings:
antibloom agents, 244--248
confectionery, 236
lecithin, 237-242
crystallization, 248--249
diglycerides, 248
fat, crystallization of, 244
milk, 241, 243
monoglycerides, 248
plastic viscosity, 237
polyglycerol polyricinoleate,
PGPR, 242-244
polysorbate 60, 24 7
sorbitan:
tritearate, STS, 246--24 7
monostearate, 24 7
synthetic lecithin, 240

Index

viscosity, 8, 237, 241, 243, 248-249


viscosity, apparent, 237
yield value, 237
chromatography/mass spectrometry, 2
circular dichroism, CD, 96
clathrate complex, starch, 68
coalescence, 95
cocoa butter:
antibloom agents, 248--249
crystallization rate, 246-24 7
polymorph, 245-247
coffee creamers, 195
column chromatography, 40-42
competitive adsorption, 167
hydrophoblic interaction, 167
critical micelle concertration, 168
ionic surfactants, 168
nonionic surfactants, 168
compound coatings:
antibloom agents, 248--249
gloss, 248
crystallization, 248--249
viscosity, 248-249
confectionery emulsifiers, 235-236
chocolate, 235, 236
compound coatings, 236
sugar, 235
concentrated milk, 199-202
production of, 201
products, 199
congeal point, 53
critical micelle concentration (erne),
148,167
cream, 174
fat globule coalescence, 183
destabilized, 187
recombined whipping, 185-186,
186
stabilization, 187

291

whipped, whipping, 183-187


cream liqueurs, 191-195
composition of, 192
cream stability, 192-195
manufacture of, 193
production of, 191
shelf life, 194
cream, coffee whiteners, 195197
recombined, 195-196
formulation, whiteners, 196
cream icings, 232
citric acid esters, monoglycerides,
266
cubic phase, monoolein/water, 131
cubic monoglyceride phase, 132
crystal size of shortenings, 216
cryo scanmng:
ice cream, 177
whipped cream, 184, 188
crystals (see also crystallization):
fat, bakery products, 217
fat, margarine, 266
fat, effect, emulsifiers, 266-267
crystallization:
effect on margarine, 267
fat, confectionery, 239
fat, chocolate, 248
fat, margarine, 265
cytochrome c, 102, 123, 126, 130

0
dairy applications, emulsifiers, 202
decaglycerol decastearate, 25
decaglycerol tristearate, 25
derivatives, bakery products, 217
diacetyl tartaric acid esters of:
monoglycerides (DATEM)

292

Index

diacetyl tartaric acid esters (continued)


esters, 31, 32, 44, 59,228,

233,249,266
differential scanning calorimetry

(DSC),53,87,96,131
diglycerides, 6, 15, 23, 25, 33, 40,

41,49,55,60,182,198,
200,204,212,227,240,
249,250,268,274,279
hydroxyl values, 23, 24
interesterification process, 15, 17
diacetyl tartaric acid, 31
dimyristoylphosphatidic acid, 126
dioleoyl PC, PE, 160
dioleoylphosphatidylserine, DOPS, 122
dioleoylphosphatdylcholine, 125
dioleoylphosphatidylethanolamine, 125
dipalmitoylphosphosphatidic acid, 121
diphosphatidylglycerol, 122
dipalmitoylphosphatidic acid, DPPA,
123
dipalmitoylphosphatdylcholine, 126
distearoylphosphatidic acid, 121, 123
DMPG, 130
dodecyl sulfate, SDS, 78
DOPG, 130
DTAB, adsorption, 100

E
egg yolk, 123
electron microscopy, 2, 266, 269, 270
ellipsometry, 121, 171
emulsifiers:
anionic, 219
analytical methods for, 39
baked foods, 211-216
bakery products, role of, 212-214
breakdown, process of, 95

bilayers, 154
binary mixture, liquid-crystalline,

150
carbohydrate interactions, 67
l3C chemical shifts, 61
chocolate confectionery, 235, 236
classified as, l l
classification, 161
coalescence, 6, 176
color of, 50
compound coatings confectionery,

236
consistency, 54
crumb softeners in bread, 70
dairy applications, other, 202
determination of, 40--52
destabilization, whipped toppings,

189
dispersion, stabilize, 154
emulsifier/starch-complex formation, 69, 79
enzyme synthesis, 286
fat, crystal structure of, 180
flocculation, 6
food additives, 2-5
functionality, 7-9
future trends, 281-286
hexagonal phase, 149
high-melting, 148
ice cream, 175
instrumental methods, 54--62
interaction, ingredients, 284--285
interfacial viscosity, 154
lactoglobulin, 114
lamellar phase, 149
margarine, function of, 267-270
melting point of, 52-54
mesophases, 284--285
micelles, 149, 154

Index

molecular solution, 149


nonchocolate confectionery, 249
nutritional studies, 282
phase diagrams, 153-156
properties of, 96, 153, 154
properties, physical, 52
polyhydric, 219
polysorbate 60, 219
processing aids, 252-253
shortenings, 211-213
solution properties of, 149
solubilization sequence, 149
sorbitan, icings, 219
specification for, 62
starch, characterization of, 85
starch-complexing, 77-78, 83
structure, S--7, 61
sugar confectionery, 235, 249
surface activity, 147-149,230
surfactants, 212
synthetic, 1
testing and surveillance of, 283
regulatory status, FDA, EEC, 215
whipped toppings, 190
emulsifier classification:
geometry of molecule, 164
homogenization role, 165
hydrophilic/lipophilic balance,

163-164
layers for, 170
phase inversion, 162
solubility, 161
emulsifier interaction, bakery components, 221
emulsifier surface, 167-169
emulsion stability, 154--155
emulsion phase diagrams, 154--155
entropy, gain of, 148
enzymatic synthesis, food, 285

293

enzymolysis in starches, 81
ethoxylated esters, 11, 26, 169, 266
sorbitan monostearate, 26
sorbitan monoolate, 27-28
sorbitan tristearate, 28
mono- and diglycerides, 28
surfactants, 169
preparation of, 28--29
polyoxyethlene (20) sorbitan, 28
ethoxylated fatty alcohols, 266
ethoxylated monoglycerides, 228
ethoxylated nonyl-phenol and xylene,

153
evaporated milk, 201
European Economic Community
(EEC), 40
baked goods in, 214
extruded snacks/cereals, 232

F
fat content, solid, SFC, 180
crystallization:
margerine, 263-267
polymorphic, 263-265
whipped toppings, 189
fat:
crystals of:
bakery products, 217
margarine, 266
effect of emulsifiers on,

266-267
crystallization:
confectionery, 239
chocolate, 248
margarine, 265
fat, yellow, consumption, 257
fat-free baked products, 232

294

Index

fat plasticity, 272, 274


fat spreads, 274
stability of, 275-276
spreadability of margarine, 277
fatty acids, 12
composition of, 264
lactylic esters, 19
metal salts of, 19, 20
melting points of, 265
soaps, 51
sorbitan esters, 58
fibrinogen, adsorption of, 108
flocculation, 95
fluorescence recovery after photobleaching, FRAP, 113
fluorescence microscopy, 126
Food and Drug Administration
(FDA), 40
regulatons of, 40
baked goods, 214
GRAS status, DATEM esters,

32
hydroxyl, 26
propylene glycol, 18
sorbitan monostearate, 26
succinylated monoglycerides,

30
Reichert-Meiss! value, 34
form stability, bakery products, 217
Fourier transform infrared spectrosopy, FTIR, 59
food emulsifiers, solution properities,

155-156
free fat acids, 60
measuring, in olive oil, 60
freeze fracture, margarine, 270
fruit acid esters, 11, 31-33
tartaric acid-derived esters, 31
fudge, 251-252

G
gas-liquid chromatography GLC), 46,

55-57
Gibbs effect, 110
performed on lipids, 55
GLC, 56
gliadin, 132
glyceryl monostearate (GMS), 86,

197
gelatin, 134
gelatinization, 78, 89
gelatinization endotherm of starches,
88
globalization, food industry, 282
globular proteins, interior, 97
hydrogen bonding, 98
hydrophobic interreaction of, 98
van der Waals interaction, 98
gluten structure, 215

gluten proteins, 224


glycerollactopalmitate (GLP), 191, 250
glycerol monostearate (GMS), 178,

180-18,189,236,248,
250,251,252
glycerol monooleate, 189
GRAS, 2-3, 15, 26
monosodium phosphate, 36
status of, 32
gums, 251

H
helix starch, structure of, 79
hexaglycerol dioleate, 25
hydroxyl value, 48
high-performance liquid chromatography (HPLC), 47, 52,57-58
detectors of, 58

Index

separations of phospholipids, 58
homogenization, 166
hydrated distilled monoglycerides, 227
hydrodynamic interactions, 166
hydrophile/lipophile balance (HLB),
6-8,151,163-164,169,
220,236
hydrophilic, 5, 11, 100
hydrated lime, 17
hydration:
force, emulsion droplets, 155
repulsion, monopalmitin, 155

295

J
jellies, 251
ACI values, 252

K
kosher, 5, 13
Krafft temperature, 148

ice cream, 95, 174-183


aging of, 178
composition of, 175
coalesence stability of, 176
cryo scanning of, 177
emulsifiers, 175, 178
fat globles in, 179
fat crystallization in, 180-182
interfacial viscosity, 177
manufacture of, 175
orthokinetic stability, 177
inclusion complexes, starch, 68
interesterification process, 17
interfacial tension, role, 165-166
in bakery products, 217
iodine
dilation value, margarine, 272
melting point, fat blends, 272
value, 45--46
binding of starches, 77
ionic surfactants, 100
isotherm adsorption, 100
isotherm surface tension, 115
IUPAC, 56

lactic acid, 236


analyses of, 49
water-in-soluble combined,
(WICLA), 22
esters of:
glycerol, 22
monoglycerides, 187, 248
propylene glycol, 22
lactalbumin, 102, 123, 125, 130, 185
lactoglobulin, 102, 104, 106, 109,
113-114,117, 118,
120-121,122,124,125,
126,130,133,134,185
lactylated esters, 18-23, 41
hydroxyl group reaction, 19
fatty acids of, 19
metal salts of, 19, 20
preparation of, 19-23
lactylated monoglycerides, 20-22,
23,44,227,230
preparation of, 20-22
lamellar liquid-crystalline phase,
emulsifiers of, 153
lamellar phase, monoglycerides, 154
Langmuir-Blodgett technique, 121
layer adsorption, 167, 169, 171

296

Index

layer adsorption, (continued)


surface, protrin, 171
phospholipid, 171
ellipsometry, 171
lecithin, 42, 50, 60, 120, 155-161,

173,199,202,220,223,
228,231,233,236,
238-239,248,250,253,
268,274
phosphorus, procedures for, 51-52
phosphatidyl, 171
phosphatidylcholine content in,

60,155
soy lecithin, 51, 239
synthetic, 240
viscosity, 54
linear molecules, starches, 68
lipid emuisifiers, liquid-crystalline,

156
lipid fractions, topping emulsions,

190
lipid materials, effects of 69, 96
lipid/protein interaction, 122
lipid structures, 129, 133
lipids in baked foods, 225
lipophilic, 5, ll
lipolytic emzymes, 133
liposomes, 84
liquid chromatography, 42
liquid-crystaline mesophases,
whipped toppings, 191
liquid margarine, 278-279
low-fat spreads, 274-278
low molecular weight (LMW), 96
lysolecithin, 84
lysophosphatidyl cholines (LPC),

119,159,283
lysophosphatides (LPC), (LPE), 156,

158
lysozyme, 102, 104

M
Marangoni effect, Ill
margarine, l, 255, 258-279
emulsifiers, 267-270
liquid margarine, 278
low-fat spreads, 274-276
market share, United States, 261
oil-in-water (0/W) spreads, 7, 278
phase inversion, 277-278
processing, 270-27 4
puff pastry margannes, 272-274
plasticity, 272-427
reduced fat spreads, 6, 274
spreadability, 277
structure, 262
fat crystallization, 263, 265-266
fat melting points, 272
polymorphism, 263-265
oils and fats, 263-426
melting points, 265
raw materials, 262
water-droplet distribution,

268-269
water-in-oil (y//0) emulsion, 7,

268,274,277-278
mass spectrometry, 60
mayonnaise, l
melting point, 12, 52-53
melittin interactions, 125
Mettler dropping point, 53
micelles, 84, 148, 165
microemulsion process, 37
milk, l, 173
composition of bovine milk, 174
concentrated, 199-202
fat globule membrane (MFGM),

173,184,185
proteins, 122, 135
recombined, 199-201

Index

shelf life, 200


minarine, 259
moisture, 50
molded products, 175
molecular aggregation, 165
molecular distillation, 15, 17
monoglycerides, 3, 6, 8, 11, 13-15,

21-22,25,40-44,55-57,
59--61, 134, 149,
154--156, 178, 182,
198--200, 202, 212,
217-219,227,230--1240,
248--250,252,268--270,
274,279
acetic acid esters of, 187, 236,
249,252-253
acetylated, 33, 44, 60
preparation of, 33-36

297

yeast-raised bakery products, 217


monoglyceride citrate, 33
monomargarine, 55, 56
monoolein, 130, 156
monopalmitin, 155, 224
monostearate, 231
cubic, water phase, 131
mouthfeel, 249
myelin protein, 125

N
near infrared reflectance (NIR), 46,

48,59
nougat, 250
nuclear magnetic resonance (NMR),

2,59,60

citric acid esters of (CITREM),

266,270,279
diacetyltartaric acid esters of
(DATEM), 3, 31-33, 41,

44,59,228,249,252,266
distilled, 22, 30, 233
distilled saturated, 156
distilled unsaturated, 156
ethoxylated, 231, 250
lactic acid esters of, 20--22, 44,

187,230,236,248
phase behavior, 178
phases of:
cubic phase, 156
lamellar phase, 156
reversed hexagonal, 156
phosphated, 3, 11, 236
propylene glycol esters of, 60, 187
stearyl monoglyceride citrate, 33
succinylated (SMG), 30, 213, 227,

278,231
tartaric acid esters of, 197, 236

0
oil-continuous emulsions, 154
oleic acids, 11-13, 256
oleomargarine (see margarine)
oleostearine, 256
organic acid esters, 156
diacetyl tartaric acid, 156
monoglyceride ester, 156
orthokinetic stability, 177
Oswald ripening, 95
olalbumin, 102

p
packing parameter, 165
palmitic acids, 11-13, 256
palmitoyloleoylphosphatidylcholine,

123

298

Index

palmitoyloleoylphosphatidylglycerol,

123
para-K-casein, 124
peanut butter, 7
penetrometer, 54
pentosan, 214
peptide chain, 97
peroxide value, 46, 4 7
phase diagrams, 153, 166
emulsion stability, 154
lamellar liquid crystalline phase,

153
water and dioleoylphosphatidylcholine, 157
phase inversion, concept, 162
phase inversion temperature (PIT),

162
phosphated esters, 36
monosodium phosphate, 36
preparation of, 36
phosphatides, mixtures of, 158
phosphatidic acids, 122, 158, 240
dioleoyl, 159
lysophosphatidylcholine (lyso PC),

159
phosphatidylglycerol, 135
phosphatidylcholine (PC), 52, 57-61,

122,130,155,157-159
dimyristoyl, 159
dioleoyl, 159
dipalmitoyl, 159
distearoyl, 159
egg, 159
soybean, 159
transition temperature for, 158
phosohatidylserine, 122
phosphatidylethanolamine (PE), 52,

60, 61, 122, 130,


156-159,173
dioleoyl, 159

dipalmitoyl, 159
soybean, 159
phosphatidylinositide, 173
phosphatidylinositol (PI), 122, 156,

158,159
soybean, 159
phosphatidyl serine (PS), 156, 173
phospholipids, 41, 42, 52, 57-61,

159,166,171,173,186,
200,202
liquid-crystalline phases, 159
phosphoric acid, 15, 17
phosphorus pentoxide, 240
photobleaching, fluorescence
recovery, 113
polar lipid:
classes, 127
cubic phase, 127
lamellar phase (LJ, 127
liquid crystalline phase, 127
hexagonal phase, 127
hydrophobic interaction, 127
micelle concentration (cnc ), 127
phase behavior, 126
surfactants, 127
structure, 128, 129
plasmalogens, 173
plasticity, 5
polyglycerate 60, 251
polyglycerol, 59
polyglycerol esters, 3, 11, 23, 54, 58,

59,213,233,249,266,
270
of stearic acid, 248
preparation of, 23-25
polyglycerol polyricinoleate (PGPR),
8,56,236,242-244
polyhydric emulsifiers, 219
polyglycerol esters, 219
sucrose esters, 219

Index

polylactic acid, 20
polymerization, 242
polymorphic form, 245, 246
polyoxyethylene (20), sorbitan monostearate, 28, 29, 114, 115,

118,156,182,220,232
polyoyl, 56
polyphosphates, 198
polysorbates, 3, 29, 59, 61, 178, 182,
219,236,250
polysorbate 60, 29, 213, 220, 227,
230,233,236,247-248,
250,252
processed cheese, 197-199
manufacture of, 198
programmed temperature x-ray diffraction, 53
propylene glycol, 17
propylene glycol esters, 15, 17, 23,

55,232
propylene glycol lactates, 23
propylene glycol monoesters

(PGME), 11, 17, 18,


22-23,213,230-232,
236,249
prepartion of, 15
propylene glycol monostearate

(PGMS), 15, 189-191


proteins, 162, 69, 71, 89
adsorption, 169
desorption, 189
strenghthening, 214
protein complexes in baked foods,

224
protein/emulsifier:
interactions, 95-135
food applications, 132
lipolytic enzymes, 132, 133
lipid structure, 133
protein stability, adsorption, 97

299

protein, lipid:
interactions, 126
monolayers, 125
protein hydrolysates, 149
protein/phospholipid interactions, 120
dispersed systems, 120, 121
solid surface interactions, 121
protein stability, 97
protein/surfactant interactions, 98--119
adsorption:
binding to, 107
competive, 111
cooperative, 107
isotherm, 99
mixtures of, 106
adsorbed:
surface properties, 103
complex formation, 105
replacement of, 105
binding, protein and emulsifiers,

113
coadsorption, 112
dissociation constant (Kd),113
emulsifier properties, 134
foam, emulsion stabilization, 110
hydrophobic surfaces, 103
hydrophilic surfaces, 103
Krafft temperature, 103
liquid air, 110
liquid/liquid interfaces, 110
lactoglobulin, 117
electrostate repulsion, 103
monolayer of, 99
nonionic surfactants, 106
interactions, solid interfaces, 106
ionic surfactants, 99, 106
solid surfaces, 98
solution influence, 107
micelles, critical concentration of,

99

300

Index

protein properties of, 102


surface tension, llO
protein/phospholipid interactions,

122
protein/protein interactions, ll2
puddings, 77
puroindolines, ll9

R
refractive index, 54
regulatory agencies, 40
Reichert-Meiss! value, 34, 49
reverse osmosis, 201
reversed aggregates, 165

s
saponification number, 26, 4 7
sauces, 77
Scatchard equation, ll6
shelf life, 200
milk, 200
salad dressings, 2
shortening, 2ll-216
emulsified, 2ll-213
fluid, 230
melting point, 216
nonemulsified, 213
plasticity, 216
solid-fat index, 216
sodium caseinate, 192
sodium dodecylsulfate, SDS, 98
sodium hydroxide, 17
sodium lauryl sulfate (SLS), 78, 134
sodium hydroxide, 17

Sodium caseinate, 192


sodium stearoyl fumarate, 197, 213,

221,231
sodium stearoyllactylate (SSL), 20,

41,42,44,156,197,203,
213,219,221,224,227,
228,230-232,266,283
softening point, 53
solubility, 161
Bancroft rule, 162
sorbide, 26
polyoyls, 26
sorbitan, 26
polyoxylated (20), 231
monostearate (SMS), 26, 56, 219,
230,231,247,248,250,
252
tristearate (STS), 56, 246, 24 7,
266
sorbitan esters, ll, 26, 162, 164,
169,220,233,266
ethoxylated, 26, 28, 266
production, 26
polyxyethylene (20), 156
sorbitan monooleate, 156
sorbitan monostearate, 156, 247
sorbitan stearate, 156
soy lecithin, 8
soybean phospholipids, 161
phase diagram, 161
spectroscopy, 59
sphingomyelin, 125, 173
spread ratios of cookie dough, 231
spreadable fats, 259, 260
starch:
complexes, in baked foods, 222
complexing, 77, 214
differential scanning calorimetry, 85

Index

inclusion complexes of, 68


iodine binding capacity, 77
composition of, 68
gelatinization of, 85, 86
temperature, Tlf Tm, 89
glass transmission, 89
crystallite melting, 89
granule, 82--83
lysolecithin, 85
nuclear magnetic resonance, 85
properties:
amylose helix, 82
effects of lipid materials, 69
fatty acids, 81
gelatinization, 77
module, 78
monoglyceride binding, 82
pasting, 70
retrogradation, 79, 80, 223
viscosity, 70, 77, 79
saturated fatty acid monoglycerides, 224
types of, 83
viscosity profile, 83
emulsifier complexes, 77, 78, 79,

83,85-90
electron spin responce, 85
infrared spectroscopy, 85
pH effects, 83
temperature, 83
paste gelation, 79
unsaturated fatty acid monoglycerides, 224
waxy, table of, 75
starch/emulsion complexes:
V-type x-ray diffraction pattern, 69
statistical experimental design, 8
fractional factorial, 8

301

response surface methodology


(RSM), 8
staled bread, 80
stearic acids, 11-13, 26, 256
stearoyllactylate, 213
succinylated esters, 11, 29
preparation of, 29, 30
sucrose esters, 3, 11, 42, 58, 59, 182,

213,227,236
preparation of, 36--38
sucrose monopalmitate, 231
sucrose monostearate, 231
sucrose distearate, 231
sucrose monostearate, 266
sucrose polyesters, 233
Olestra, 233
surface activity, 148, 167
components, 167
polar interactions, 148
surfactants, 5
surface tension, emulsifiers, 147,

227
surfactants, 168, 199, 201, 212
anionic, 199, 201, 224
cationic, 199
cetyl-trimethyl ammonium
bromide (CTAB), 199
ionic, 99, 100, 168
interaction of, 101
lecithin, 199
molecules, arraingments of, 101
nonionic, 168, 201, 224
properties, 102
sodium dodecyl sulphate (SDS),

199,201
solidlaqueos interfaces, 99
sucrose ester, 223
zwitterionic, 199

302

Index

T
tetraglycerolesters, 156
monolaurin, 156
tetraglycerol, 156
thin film stabilization, 111, 112
tocopherols, 50
toffee, 250
triglycerides, 2, 12-5, 17, 204, 240,

263
glycerolysis of, 204
triglycerol, 23
triglycerol monooleate, 248
triglycerol monoshortening, 250
triglycerol monostearate, 248, 252
triglycerol monoshortening, 25
triglycerol monostearate polyglycerol
ester, 25
tripropylene glycol monoesters, 18

u
ultrafiltration, 201

vitamin fortification, 261


Vroman effects, 108

w
water-continuous emulsions, 154
water-insoluble combined lactic acid

(WICLA), 49
waxy starch table, 75
wet chemical analysis, 43-50
titration, 43-49
wheat flours, 214
whipped cream, whipping cream,

183,185
electron micrograph, 188
recombined, 185, 186
whipped toppings, 187
emulsion destabilization, 189
fat crystallization, 189
powder composition, 187
protein desorption, 189
Wiley melting point, 53

v
van der Waals interaction, 98
vesicle bilayers, 131
viscosity, 8, 54,177,237,241-244 ,

248
apparent, 237
interfacial, 177
plastic, 237, 243, 244

yeast-raised products, 225


crumb-softening, 225--227
dough conditioning, 225
dough strenghthening, 225, 226
emulsifier functionality during
processing, 227
yield value, 237
yoghurt, 202