FT 131

JAN 23

FOOD CHEMISTRY
- post harvest handling of fresh produce
- food formulation
- food processing
- storage
- proper investigation/research: facts derived from the study of one food or model system

Examples:
1. EFFECT OF FOOD ADDITIVE:
- antioxidant in preventing rancidity: stability of sweetened-dried young coconut meat with tocopherol
randomly selected from 1 batch will enhance our understanding of the stability of the rest of the
products and of the future products.
- synthetic sweetener as substitute for sugar in polvoron: 60% substitution of sugar significantly
reduced calories of the product without affecting sweetness (perceived). Results can be used in
modifying similar products or other products.
2. POST-HARVEST CHANGES IN FRUITS
- biochemical/chemical changes (ripening) in saba banana at 15C; results will enhance our
- biochemical/chemical changes in solo papaya under MA (Modified Atmosphere) storage

4 COMPONENTS OF ANALYTICAL APPROACH

1. Determination of properties that are important characteristics of safe-high quality foods
2. Determination of chemical and biochemical reactions that influence the loss of quality and/or
wholesomeness of foods
3. Integrating approaches 1 and 2 so that one understands how the key chemical and biochemical
reactions influence quality and safety of foods
4. Application of the knowledge gained in no. 3 to the various situation encountered during the
formulation, processing and storage of foods

SAFE-HIGH QUALITY FOODS

> safety first requirement to be satisfied by any food (fresh or processed)
> free from hazards (biological/microbiological, chemical, physical) at the time of consumption
Examples:
1. canned products, low acid foods, pH>4.5, absence of Clostridium botulinum it toxins
2. drinking water, no heavy metals, e.g. Hg

> quality-sensory and nutritive value: influenced by composition and properties of components
> sensory attributes color, aroma, flavor, taste, texture
> chemical and biological reactions: desirable or undesirable
Desirable enhance quality of food; e.g. tenderization of meat during aging
Undesirable deterioration of food; biochemical/chemical reactions resulting to loss of quality
attributes specific to the food.

Reactions Results

Hydrolysis of lipids ffa react with proteins: change in flavor

and texture
Hydrolysis of polysaccharides sugar released react with protein or aa:
change in texture or flavor
Hydrolysis of pectin softening, loss of crispness

Association of molecules change in texture, gel formation

MECHANICAL DAMAGE
- physical changes which may induce biochemical/chemical reactions
Examples:
1. Bruising of fruits, vegetables, root crops
1
 2. Heating
3. Low temperature, refrigeration, freezing

1. Bruising of fruits, vegetables, root crops

- may release enzymes which catalyze biochemical changes such as
a. browning discoloration phenolases or polyphenol oxidase (PPO)
b. changes in texture tenderization or softening due to destruction of cell walls, cell membrane by
hydrolases
c. changes in flavor loss of formation of flavor compounds

2. Heating
- cause protein denaturation: change in texture, flavor, etc.

3. Low temperature, refrigeration, freezing

- chilling injury, freezer burn

PRESERVATION OF FOODS

A. Short term preservation extend freshness of fruits, vegetables, seafoods, beef, pork, chicken
(poultry), etc.
- refrigeration, modified atmosphere packaging (MAP), controlled atmospheric packaging (CAP)
B. Long term preservation canning, drying, freezing

In the selection of the method consider the following: compostion, components of food, post harvest, post
mortem biochemical/chemical changes in food (may occur immediately after harvest/death, during
processing or storage, thus there is a need to study components of food, macro components, and micro
components).

PIGMENTS AND OTHER COLORANTS

Color one of the most important quality (sensory attribute of food)
- influence acceptability; particularly color associated with/to specific food items or quality
- influence palatability; index of maturity, e.g. fruits, index of freshness, e.g. meat; influence extent/type
of processing
- inherent in food
- (colorant) added to food to enhance its sensory quality (not to cover faulty processing technique)

Color of food:
1. Pigments inherent in food* - fresh produce
2. Colorants (added) processed foods
*preserved during processing/preparation?

PIGMENTS
- normal constituents of cells or tissues of plants or animals that impart color
- in fruits, leaves, other parts of plants, animal tissues, blood, fish, shellfish, other seafoods
- in harvested fruits/vegetables and muscle foods, pigments also include compounds/substances
formed on heating, storage or processing
- also synthesized by some microorganisms

JAN 30
COLORANT
- general term which refers to any chemical compound that imparts color
- food additives, i.e., intentionally added
- can be a natural pigment extracted or prepared (e.g. annatto, turmeric, carotenoids, anthocyanins)
from a plant source or it can be synthetic
- regulated particularly the synthetic food colors (subjected to toxicological tests prior approval to use)

OTHER FUNCTIONS OF PIGMENTS

- in live plant/animals
energy receptor chlorophyll
2
 carrier of oxygen
protectants against radiation some carotenoids
- post harvest
some are considered as neutraceuticals, antioxidants which prevents certain diseases, e.g. cancer

CHLOROPHYLL
- belong to the group of tetrapyrrole compounds
- all classes of photosynthesized porphyrin pigments
- green pigments involved in the photosynthesis of higher plants

TYPES
1. Chlorophyll a in all plants, algae and cyanobacteria
2. Chlorophyll b in plants and green algae only
3. Chlorophyll c in photosynthesis chromista and dinoflagellates
4. Chlorophyll d in red algae together with type a

QUESTION: What are the types of interest to food chemists?

- in higher plants ratio of a to b ~ 3:1

STRUCTURE:
- porphyrin ring with 4 pyrrole group and Mg
a Mg chelated tetrapyrrole structure with methyl substituents; with phytol** group esterified to
propionate at the C-7 position
**phytol a 20C monosaturated isoprenoid alcohol

Type R group*

Chlorophyll a CH3 (methyl)

Chlorophyll b CHO (formyl (aldehyde))

**STRUCTURE
- Location:
Chloroplasts plastid bodies where chlorophylls are located
Grana smaller particles with in chloroplasts composed of lamellae where chlorophyll molecules are
embedded
Stroma between grana

- Occurrence in plants associated with lipids, proteins and lipoproteins bound to these molecules
How?
Held together in a monolayer by:
- mutual attraction
- affinity of phytol tail for lipids
- affinity of the hydrophobic planar porphyrin ring for proteins

3
- Properties: Physical and Chemical
Physical, solubility in various solvents:
S Soluble
I Insoluble
SltS Slightly soluble
AlmI Almost Insoluble

Alcohol Ether Benzene Acetone Petroleum Water

Ether
chlorophyll a S S S S SltS I

pheophytin a S S S S SltS I

chlorophyll b S S S S AlmI I

pheophytin b S S S S AlmI I

Chlorophyllide S
a/b* *In General Insoluble in Oil
pheophorbide S
a/b

Chemical Properties
- chemical changes in structure of chlorophyll
- alterations in the structure are manifested through changes in color of disappearance of color

Factors which influence changes:

- temperature (heating, cooking)
- pH (acidity)
- light (sunlight, artificial light, packaging)
- etc.

Alterations/changes in the structure of chlorophyll

1. Isomerization
2. Pheophytinization
3. Removal of phytol group from chlorophyll
4. Removal of phytol group from pheophytin
5. Removal of Mg from chlorophyllide and its replacement with H
6. Formation of pyrophephytin
7. Formation of pyropheophorbide
8. Reaction of side groups
Ex. Oxidation of isocyclic ring allomerized chlorophyll
9. Rupture of tetrapyrrole ring
- colorless end products
- 2-5 common reactions

Chemical changes
1. Isomerization of chlorophyll by inversion of carbomethoxy group at C10
- may occur during heating in the presence of organic solvent
Ex.
chlorophyll a chlorophyll a
These isomers have stronger A on C18
chlorophyll b chlorophyll b
Reverse phase HPLC column
*observed in spinach leaves when heated at 100C for 10 min, 5-10% conversion

4
 2. Pheophytinization removal of Mg++ and its replacement with 2H+
-Mg++, +2H+
chlorophyll pheophytin
bright green dull olive brown
compare susceptibility of chlorophyll a with chlorophyll b to pheophytinization
- chlorophyll a is more susceptible; chlorophyll b is more stable

3. Removal of phytol group from chlorophyll

-phytol
chlorophyll chlorophyllide
bright green chlorophyllase bright green (more intense)

presence of Mg++ - bright green color

4. Removal of phytol group from pheophytin

-phytol
pheophytin pheophorbide
olive brown olive brown

5. Removal of Mg from chlorophyllide and its replacement with H

-Mg++, +2H+
chlorophyllide pheophorbide
bright green olive brown

6. Formation of pyropheophytin
-CO2CH3* at C10, +H at C10
pheophytin pyropheophytin
*CO2CH3 carbomethoxy

7. Formation of pyropheophorbide

-CO2CH3* at C10, +H at C10

pheophorbide pyropheophorbide

SUMMARY OF CHEMICAL CHANGES

-Mg++, +2H+
chlorophyll pheophytin
(bright green) (olive brown)

-phytol -phytol -CO2CH3* at C10,

+H at C10

-Mg++, +2H+
chlorophyllide pheophorbide
(bright green) (olive brown)

pyropheophytin
CO2CH3* at C10,
+H at C10

pyropheophorbide

5
FEB 02
CHLOROPHYLLASE an esterase; cleavage of phytol from chlorophyll and pheophytin (derivative of
chlorophyll)

- specific to porphyrins with carbomethoxy groups at C10 and H at C7 and C10

- active in solution with watein nr, alcohol, acetone
- optimum temperature, 60-82.2C, losses activity at 100C

Temperature in raw fresh vegetable? 28-34C

Favorable for chlorophyllase activity? No

In fresh leaves, chlorophyllide is formed only when chlorophyllase is heat activated during post harvest
e.g. during blanching

EFFECTS OF FOOD HANDLING, PROCESSING & STORAGE ON CHLOROPHYLL

- degradation, rate of extent differ depending on the type/degree of treatment to which the food item
was subjected
(recall the alterations/changes* in chlorophyll
*induced

FACTORS WHICH CAN LEAD TO CHLOROPHYLLL DEGRADATION

1. Heat treatment
2. Exposure to light
3. Enzyme activity
4. Irradiation (before storage)
5. Fermentation
6. Added ingredients

1. HEAT TREATMENT: Blanching, thermal processing

- consider stability of a and b (more stable) of chlorophyllase (opt. at 60-82.2C)
Blanching: recall temperature and methods
Milder compared to thermal processing
Temperature: between 60-82.2C will enhance chlorophyllase activity
: 100C (short time) loss enzyme activity but reactivated during storage
Effect of blanching: green color enhanced or retained
Why? Formation of chlorophyllide
Prolonged storage: degradation of either frozen products or dried products

Higher temperature, longer time more degradation

Kinetic sequence:
Chlorophyll pheophytin pyropheophytin
green olive-brown

Why is chlorophyll b more stable than b?

Stability of chlorophyll b is attributed to the e- with drawing effect of C3 formyl group
(recall structure)

- increase in +charge on the 4 pyrrole Ns reduces the equilibrium constants for the formation of the
reaction intermediate

Factors which influence the effect of heating:

1. pH
2. chloride salt of Na, Mg and Ca
3. detergents cationic and anionic

pH of the tissue
pH 9.0 very stable during heating
pH 7.0 unstable during heating

6
 Q.1. What is the pH of green leafy vegetables, green beans and other green vegetables?
A. Most leafy vegetables have pH >3.0, hence may remain stable during cooking/processing
Q.2. Why is it that many green leafy vegetables loss their bright green color on cooking, particularly
with longer cooking period?
A. There are factors that influence effect of heating which include pH and cooking. Cooking
(long exposure to heat)/processing destroys structure of cells, cell walls, membranes, etc.,
which can result to: 1) release of acids (inherent) and; 2) formation/production of acid.
Example: spinach, peas, green beans observed to release/produce 10 acids.
Acids implicated:
1. Acetic
2. Pyrrolidone carboxylic acid (major cause)
3. Oxalic acid
4. Malic acid
5. Citric acid
6. Succinic acid
7. Fatty acids from fat hydrolysis
8. Other acids

Chloride salts of Na, Mg and Ca

- observed to have reduced chlorophyll degradation.
How?
- It is because of electrostatic shielding effects of salts where cations were proposed to neutralize
the negative (-) surface charge of fatty acids and proteins on the membrane of chloroplasts resulting to
the reduction attraction of H+ to the membrane surface.

Na+ or Ca2+ or Mg2+ vs H+ H+ (from acid)

fa- proteins-
chloroplast

Detergents
- cationic detergents (+) repel H+ at the membrane surface
- limit H+ diffusion into the cell

2. EXPOSURE TO LIGHT
- cause photodegradation of chlorophyll* : loss of desirable green color
*occurs:
1. during senescence
2. during storage
3. when extracted
4. when there is cell damage, e.g. during processing

Occurrence in healthy plant cells? Same degree of degradation?

- Minimal or may not occur in healthy plant cells

Healthy plant cells chlorophyll is protected from destruction by light due to the presence of
surrounding carotenoids and other lipids

7
SUGGESTED MECHANISM OF PHOTODEGRADATION

opens* fragments
tetrapyrrole lower mol. wt. compounds

*opening of methine bridges oxidized linear tetrapyrrole

Lower molecular weight compounds

1. glycerol major breakdown product
2. lactic acid
3. citric acid
4. succinic acid when formed, loss of color,
5. malonic acid i.e. pigments
6. alanine
7. singlet oxygen can react with other tetrapyrrole chlorophyll
8. hydroxyl radicals or other porphyrin to form peroxides and
more free radicals -> total loss of color

Singlet oxygen and hydroxyl radicals can react with other tetrapyrrole chlorophyll or other porphyrin
to form peroxides and more free radicals -> total loss of color

Ex. chlorophyll + singlet O2 peroxides peroxides + free radicals

green loss of color

Similar reactions with other porphyrins and hydroxyl radicals

Prevention of the undesirable effect of light on processed foods

- proper packaging (clear packaging material vs opaque/dark-colored packaging

3. ENZYME ACTIVITY (BESIDES CHLOROPHYLLASE)

Note: Some enzymes may not directly degrade (act on) chlorophyll but products of reactions that
catalyze have been observed to degrade chlorophyll.
Example:
Lipoxygenase degrade fats/lipids and form free radicals which in turn degrade chlorophyll

lipoxygenase
lipids free radicals

chlorophyll + free radicals degradation products (undesirable color

formed
(green) (brown)

4. IRRADIATION: GAMMA-IRRADIATION
Fruits/vegetables before storage

irradiation
chlorophyll pheophytin

5. FERMENTATION
Decrease in pH due to acid formed aside from the inherent acids present

fermentation
chlorophyll pheophytin, chlorophyllide,
normal pH pheophorbide, acidic conditions, lower than normal pH

8
FEB 06
6. ADDED INGREDIENTS: ACID, LIME
Acid pH, reduction
- pheophytinization

Q. Why?
H+ ions from added acid can replace Mg++ in the porphyrin ring (Note: 2H+/Mg++

Lime increase in pH; maintain green color

- extreme alkalinity -> degrade chlorophyll, loss of color

Restoration of chlorophyll/green color

Can the structure of chlorophyll be fixed or restored?
Is it possible to restore the green color in leaves? How?

Metallo Complex Formation in pheophytin and pheophorbide (not in intact chlorophyll)

Recall: Mg++ is readily replaced by H+ under acidic conditions

Questions:
1. Can we remove H+ and add back Mg++?
2. Can another metal ion replace H+?

Recall the electrochemical series

Metal ions: Cu++, Zn++
Reactions: pheophyrin + Cu++ Cu-pheophyrin* + 2H+
favored at low pH green
*stable at low pH than at alkaline conditions

pheophyrin + Zn++ Zn-pheophyrin* + 2H+

green; stable at pH 2.0
Metallo Complex Formation
- pheorphide can undergo similar reaction

Comparison of reactivity of pheophytin and pheophorbide in metallo complex formation:

- pheophytin is less reactive than pheophorbide. Why? Size? Structure?
- compare structure (pheophytin vs pheophorbide)
- the phytol tail in pheophytin causes steric hindrance making it less active than pheophorbide

Recall structure of chlorophyll a and b

Comparison of reactivity of chlorophyll a (corresponding derivative) and chlorophyll b (corresponding
derivative) in metallo complex formation:
- chlorophyll a forms metallic complex with either metal ion faster than chlorophyll b
- the formyl (OCH3) group in chlorophyll b has an e- with drawing effect which makes the pyrrole
atom becomes more +, therefore less reactive with the metal

PRESERVATION/RESTORATION OF THE GREEN PIGMENT/ COLOR

Can a food technologist prevent chlorophyll degradation?
Knowledge about structure and properties of chlorophyll*

DEGRADATION
- mechanisms/reactions of degradation
- causes, factors which influence (flavor) degradation

*Food technologists/food chemists in controlling/preventing undesirable changes in the pigment

METHODS IN PRESERVATION/RESTORATION OF THE PRESERVATION (recommended)

1. USE OF ALKALINE SALTS (ACID NEUTRALIZATION)

- neutralize the medium,e.g. raise pH to 7.0

9
 - alkaline salts:
a. Ca + NaH2PO4
b. MgCO3 in combination with Na3PO4
c. Na2CO3
d. Ca(OH)2 + Mg(OH)2
e. Ethyl cellulose + 5% Mg(OH)2 as interior coating of cans

- a and b are not highly recommended due to softening of tissue and alkaline flavor
- d [Ca(OH)2 + Mg(OH)2]
known as Blair Process, originally used to preserve texture and crispness of some vegetables
not very successfully commercially for the retention of the desirable green color due loss of color in
less than 2-month storage. Why? Salts were not effective in neutralizing acid in the inner tissues of
vegetables at longer period.
- e. ethyl cellulose + 5% Mg(OH)2 as interior coating of cans

slow leaching
Mg(OH)2 medium, ~pH 5.0

- not very successful commercially (partial). Why?

1. increase in pH results to amide hydrolysis
glutamine or aspargines -> products with NH3-like odor
2. fatty acids* present are oxidized to products associated with rancidity
*fatty acids may be formed during heating e.g. blanching at higher pH
fatty acids -> peroxides, hydroperoxides, other products
3. struvite* formation
- occurs at higher pH, e.g. observed in peas at pH 8.0
*glass-like crystals consisting of Mg and NH4 phosphate complex (from protein and amino acid)

2. HTST (HIGH TEMPERATURE SHORT TIME) PROCESSING

- products with better retention of vitamins, flavor and color
- color retention limited to 2 months only, beyond which loss of color was observed
- heating particularly within the optimum temperature for chlorophyllase activity favors retention of the
green color

3. CONVERSION OF CHLOROPHYLL TO CHLOROPHYLLIDE

Chlorophyllase
Recall: chlorophyll chlorophyllide
opt. temp 60-82.2C (more stable than chlorophyll)

Blanching inactivates enzymes if high temperature is used (one of the purposes of blanching)
Optimum temperature for chlorophyllase activity

Compare blanching HTST vs LTST

Observations: blanching using LTST

1. 71C <-> 20 min -> better color retention in spinach
2. 65C <-> 45 min -> better color retention in spinach
3. 54C <-> 45 min -> activation of chlorophyllase

Disadvantage: color retention is not sufficient for commercialization

4. APPLICATION OF METALLO COMPLEX

Recall: Effect of Cu and Zn ions or chlorophyll derivatives, pheophytin and pheophorbide

Use of Copper Ions

- allowed in European countries under EEC (European Economic Community) to a certain types
of food product; canned foods, candies, soups and dairy are not allowed in US

10
 Commercial forms of copper metallo complexes
Copper chlorophyll = copper + pheophytin
Copper chlorophyllin = copper + pheophorbide

FAO of UN certified the use of copper metallic complexes as safe in foods provided no more than 200
ppm (mg/kg) of free ionizable copper present

Veri-green refers to blanching vegetables in water with Zn or Cu salts to raise tissue concentration
of metal to 100-200 ppm

Commercially acceptable vegetables are green

Ex. canned green peas, pigments are: Zn pheophytin and Zn pyropheophytin

Direct adding of Zn/Cu ions in canning brine had no significant effect on chlorophyll
- best process to retain green color in vegetables
- blanch in the presence of Zn ions
- increase membrane permeability by heating tissues to 60C before blanching
- provide the right pH, i.e., favors metallo complexes
use anions to alter surface change of issue
- recall effect of surface active detergents

Regreening of Thermal Processed Vegetables

- appearance of bright-green areas in the product due to the presence of Cu or Zn ions in
process solution

FEB 09
FLAVONOIDS
- subgroup under phenolic compounds
- widely distributed in plants
- associated with the yellow color in some plants; also responsible for the whiteness in some plants; also
impart the following colors: blue, purple, magenta, red, orange, due to anthocyanins
- carbon skeleton: basic C6C3C6 structure
- color depends on the substituents on rings A & B

Basic structure: phenolic compound (s) 3 benzene rings, with OH

Typical Flavonoid Structure

The Structure of a Flavonoid:

Phenolic compounds composed of three benzene rings with hydroxyl (OH) groups
Without sugar, molecule called an aglycone

11
 1. Remove the OH at (1): flavone
2. Replace OH at (1) with Zn ring: isoflavone
3. Replace the O at (2) with a H: anthocyanin
4. Replace the OH at (3) with glucose;
Remove OH at (4) ;
Remove OH at (1): glucoside
(glycoside if exact sugar is not specified)

ANTHOCYANIN
- most prevalent flavonoid
- Greek words: anthos flower; kyanos blue
- Impart a wide range of colors in plants: blue, purp[le, magenta, red, orange

Most notable investigators (Nobel prizes in Chemistry) in part of work in plant pigments
1. Robert Robinson (1886-1975)
2. Richard Willstatter (1872- 1942)

There are 250 anthocyanins in plants, content vary from about 20mg/100g to 600mg/100g fresh weight

Q. Which is responsible for the color in fruits, flowers, leaves (or any plant part)?
A. Distinctive color in the plant part: often due to a combination or a system of pigments*
*usually 4-6 pigments, one may predominate
Examples:
1. Simple System
blackberries contain primarily cyanidin-3-glucoside
2. Complex System
blueberries contain 15 pigments

Base structure: flavylium cation, a 2-phenylbenzopyrylium of flavylium salt

R1, R3 H, OH or OCH3
R2 OH
R4 glycosyl
R5 H or glycosyl

ANTHOCYANINS IN PLANTS
- exist as glycosides of polyhdroxy and/or polymethoxy derivatives of the flavylium salts
- differ in the R groups:
*no. of hydroxyl or methoxy groups
*types, no. and site of attachment of sugars to the molecule
*types and no. of aliphatic or aromatic acids attached to the sugars (some contain organic acids &
metals Al, Fe, Mg)

2 MAIN COMPONENTS OF ANTHOCYANIN (exist as glycosides in plants)

1. Anthocyanidin (aglycone, aglucone [Whittaker])
2. Sugar(s)

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HYDROLYSIS OF ANTHOCYANINS

hydrolysis
anthocyanin anthocyanidin + sugar

SUGARS:
1. glucose
2. galactose
3. arabinose
4. xylose
5. rhamnose

*1-4 most common;

*1 or 2 or 3 sugars may be present, homogeneous or heterogeneous

CLASSES OF ANTHOCYANINS BASED ON THE SUGAR PRESENT:

1. monoside
2. bioside
3. trioside

SUGARS MAY BE ACYLATED BY EITHER ALIPHATIC OR AROMATIC ACIDS:

caffeic malonic
p*-coumaric *para malic
sinapic succinic
p-hydroxybenzoic acetic
ferulic

COLOR OF ANTHOCYANINS AND ANTHOCYANIDINS

- results from excitation of the molecules by visible light
- ease of excitation depends on the electron mobility in the structure

STRUCTURE OF ANTHOCYANINS AND ANTHOCYANIDINS

- with several double bonds which are readily (easily) excited, presence of which is important for color
- hence, the type of anthocyanidin influence color or deepness of color

SIX COMMONLY OCCURRING ANTHOCYANIDINS IN FOODS

[may vary in hue (color) and intensity (deepness)]

Pelargonidin Cyanidin Delphinidin

R 1, R 3 H R1, R2 OH R1, R2, R3 - OH
R2 OH R3 H

Peonidin Petunidin Malvidin

R1 OCH3 R1 OCH3 R1, R2 OCH3
R2 OH R2, R1 OH R2 OH
R3 H

Anthocyanidin
Top to bottom: increasing redness; increasing no. of OCH3 (methoxy group) *see column 2 & 3
Left to right: increasing blueness; increasing no of OH (hydroxyl group) *see row 1

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FEB 13
Explain observation: Bathochromic changes
Deepening of hue is due to the bathochromic change (i.e., longer wavelength) light absorption band in the
visible spectrum shifts from violet through red to blue

Q. What brought about this change?

A. The auxochrome groups* - groups without chromophoric properties (colorless) but cause deepening of
the hue when attached to a molecule, i.e., anhocyanidin
*e- donating; *has bathochromic effect

AUXOCHROME
- (auxanein) to increase and (chroma) colour
- is a group of atoms attached to a chromophore which modifies the ability of that chromophore to
absorb light

BATHOCHROMIC CHANGE
*e- donating; *has bathochromic effect
In anthocyanidins:
R groups: hydroxyl, OH and methoxy, OCH3
Compare bathochromic effect of OH and OCH3
OCH3 has greater e- donating capacity than OH -> cause greater bathochromic effect
HYPSOCHROMIC CHANGE opposite change

PROPERTIES OF ANTHOCYANINS
Solubility water soluble
Note: anthocyanidin less water soluble than anthocyanins
Chemical properties readily undergo changes with pH changes
- relatively unstable, degraded during extraction, processing, storage

STABILITY OF ANTHOCYANINS
Recall: structure and substituents (subgroups) on the aglycone (anthocyanidin)

INFLUENCE OF SUBSTITUENTS ON STABILITY:

1. OH increase hydroxylation, decrease stability
2. OCH3 methylation, increase stability
3. Sugars consider no.4 type of sugar
>number of sugars increase glycosylation, increase stability
monoglucoside vs diglucoside

Monoglucoside vs Diglucoside
Examples:

Anthocyanin, pH 3.5, 50C Half-life, hr

Cyanidin-3-rutinoside 16

Cyanidin 3-2 glucosylrutinoside 26

TYPE OF SUGAR
Anthocyanin containing galactose shown to be more stable than anthocyanin containing arabinose in
stored cranberry (Starr and Francis, 1968)
Note: Substituents (1-3 [OH, OCH3, sugar]) do not react but influence marked effect on stability.

INFLUENCE OF THE TYPE OF ANTHOCYANIDIN/AGLYCONES

1. pelargonidin
2. cyaniding 1-3 anthocyanins are less stable
3. delphinidin
4. petunidin (-OCH3) 4-5 anthocyanins are more stable; increase in stability is due to the
5. malvidin (2 OCH3) blocking of the reactive hydroxyl groups
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FACTORS WHICH AFFECT STABILITY OF ANTHOCYANINS
1. MAJOR FACTORS
A. pH
Four structural forms
1. blue quinoidal base (A)
2. red flavylium cation (AH+) one form may predominate over the others depending
3. carbinol pseudobase (B) on the pH
4. colorless chalcone (C

A AH+ B C (Assumption: equilibrium exists)

low pH pH 4-6
One may predominate over the others, depending on the pH
Example:
Malvidin-3-glucoside, pH 0.6
pH 4-6: carbinol pseudobase dominates; colorless
pH <4: plavylium cation dominates; red

B. TEMPERATURE
- rate of degradation is influenced by pH, O2 and structural conformation
- structural features which increase pH stability, also increase thermal stability

highly hydroxylated anthocyanidins vs methylated, glycosylated or acylated

anthocyanidins
more stable
Example:
Half-life of the following at pH 2.8

Anthocyanins Half-life (days)

3,4,5,5,7-pentahydroxyflavylium 0.5
3,4,5,5,7-pentamethoxyflavylium 6.0
cyanidin-3-rutinoside 6.5
cyanidin 0.5 (12hr)

C. OXYGEN
- anthocyanins are unsaturated, readily undergo oxidation
- manifestations: browning of some fruits or its products

*Influence of Aw on stability against oxidation

Aw = 0.63 0.79 -> anthocyanins are more stable

*Aw of dried fruits/vegetables, if there is minimal degradation of anthocyanins during drying, such
will be preserved, hence color will be maintained

Prevention of Oxidation:
- exclusion or removal of oxygen during drying: vacuum drying, freeze drying

*Processing:
1. proper exhausting before sealing of canned/bottled products
2. processing under nitrogen atmosphere or under vacuum
3. packaging in nitrogen atmosphere (e.g. powdered grape juice)
4. vacuum packaging

D. LIGHT
- accelerates anthocyanin degradation

Photodegradation adverse effect observed in fruit juices and wines

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 Comparison of anthocyanin stability in the presence of light

acylated,methylated non-acylated monoglucosides

diglucosides digluclosides
------------------------------decreasing stability towards photodegradation--------------------------------->

E. CONCENTRATION

FEB 27
2. MINOR FACTORS

A. DEGRADATIVE ENZYMES
Two groups implicated
1. Glycosidases generally referred to as anthocyanases however substrates acted are not
2. Polyphenol oxidases limited anthocyanins hence use of term is not encouraged

Glycosidases hydrolyze glycosidic linkage

glycosidase
anthocyanins sugar + anthocyaninidin loss of color intensity*

*results from:
1. Formation of colorless products
2. Loss of solubility of anthocyanidin
-> polyphenol oxidase (PPO) oxidize/degrades anthocyanins in the presence of o-diphenol and
oxygen
-> not directly act/oxidize anthocyanins; oxidize o-diphenol and product formed reacts with
anthocyanins
Steps:
PPO
1. o-diphenol + O2 o-benzoquinone
2. o-benzoquinone + anthocyanin oxidized anthocyanin + degradation products
brown products
1 enzymatic 2 chemical

Catechol oxidase copper oxidase, catalyzes oxidation of ortho-diphenol to ortho-quinones

B. ASCORBIC ACID
- degradative motion on anthocyanins is indirect; it undergoes oxidation and products degrade
anthocyanins

Cu++
ascorbic acid ------- H2O2
cleaves pyryllium ring by a nucleophilic attack at C2
anthocyanin + H2O2 ---------> colorless esters + coumarin derivatives ---> brown precipitate*
*occurs in some juices

C. SULFUR DIOXIDE
- used in food processing*
*as preservative: 500 ppm to 2000 ppm
*as bleaching agent: higher concentration; 0.8-15%
Ex. production of maraschino or candied & glace cherries

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 Effect of SO2 loss of color

Reversible or irreversible?
Reversible: color can be restored through desulfuring , i.e., wash off SO2 before further processing

Reaction:
anthocyanin + SO2 ------> complex formation (colorless)
Suggested reaction: SO2 attacks C4

Note:
1. Some anthocyanins are resistant to bleaching; C4 is blocked or exists as dimmers linked
through C4
2. Irreversible in maraschino or candied cherries

D. METAL IONS
Anthocyanin + metal complex

Nature: metal complexes of anthocyanins give characteristics colors to flowers

In food processing, e.g., canning, metal anthocyanin complex formation may occur*
*desirable or undesirable

Observations: effect of complexation

1. Complexation stabilizes (metal protects) some anthocyanins beneficial Ca, Fe, Al, Sn, in
cranberry juice
2. Complexation results to discoloration
3. Pinking occurs in pears, peaches & lychees; undesirable

Mechanism (in complexation):

Vicinal phenolic hydroxyl groups (in some anthocyanins) sequester multivalent metal*
* Al3+, Ca2+, Fe2+, Sn2+

Note:
Not all anthocyanins have vicinal phenolic hydroxyl groups, hence metal complexation can be
used to differentiate groups of anthocyanins

Mechanism (in complexation):

Complexation, an analytical tool to differentiate groups of anthocyanins
Reagent: aluminum chloride

Anthocyanins which can complex with metals**

Cyanidin Pelargonidin
Petunidin Peonidin
Delphinidin Malvidin

**with vicinal phenolic OH, recall R1, R2, and R3 groups

Mechanism in pinking:
heat; acidic conditions metal
Proanthocyanidin anthocyanins metal-anthocyanin complex
(colorless) (pink)

E. SUGARS
Effect on anthocyanins depends on sugar concentration; beneficial or not

High sugar concentration, i.e., fruit preserves, stabilize anthocyanins

Stabilizing effect is due to the lowering of Aw

Low sugar concentrations, i.e., at concentration with little or negligible effect on Aw -> accelerate
anthocyanin degradation
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 Types of sugar -> varying effect

Fructose
Arabinose FALSor greater degradation
Lactose
Sorbose
Glucose
Sucrose MGSuc (or MSG) less degradation
Maltose

Mechanism: (not direct interaction between sugar and anthocyanin) degraded

sugar furfural
O2
anthocyanins + furfural brown compounds (discoloration)
condensation

EFFECT OF COPIGMENTATION
- either accelerate or retard depending on the situation

Protective effect of some flavonoids:

1. polyhydroxylated flavone sulfonate*
2. polyhydroxylated isoflavone sulfonate* *protective effect against photodegradation
3. polyhydroxylated aurone sulfonates*

- the negatively charged sulfonates and the positively charged flavylium cations form intermolecular ring
interactions

EFFECT OF SUBSTITUTIONS OF OH GROUP

o-diphenol
more susceptible to photodegradation

- unsubstituted or monosubstituted anthocyanins -> prone to nucleophilic attack at the C2 or C4

positions

PREVENTION OF PHOTODEGRADATION:
*Proper packaging, e.g. dark colored bottles or plastic films
Ionizing radiation degrades anthocyanins

OTHER FLAVONOIDS
- non-anthocyanin type flavonoids
- impart a range of color to plants and food
yellow: yellow flavonoids
xanthos yellow
- whiteness of some plant materials
- brown or black due to the oxidation products of phenolic groups (occur in nature)
>>classes of non-anthocyanin type flavonoids (differences is based on the state of oxidation of the 3-
carbon link)

PROPERTIES
- with light absorption characteristics
- differ due to differences in degree of unsaturation
- also influenced by the acylation and/or glycosylation
- involved in copigmentation
- metal chelators
chelation or reaction with metal; influence color formation
Ex. Luteolin + Al -------> yellow complex

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IMPORTANCE IN FOODS:
1. Color, particularly in copigmentation
Ex.
>whiteness: in onions, cauliflower, potato

>greenish: black discoloration in canned asparagus

Rutin** + Fe+++ Fe rutin complex

**Rutinoside quercetin Control: add EDTA, metal chelator

>yellow color in wax bean

Rutin + Sn++ Sn rutin complex (more stable)

>color of black olives due to the oxidation of flavonoids

Reaction occurs during fermentation and storage
Ex. luteolin 7-glucoside -------> oxidation products

2. Antioxidant properties
- ability to sequester metal

3. Contribute to flavor, e.g., bitterness, astringency

Ex. tea astringent
>contains quercetin
kaempherol contribute to astringency
myricetin glycosides

MAR 02
4. Potential sweetener
>citrus, has naringen* bitter
*a flavanone potential sweetener

5. Deodorizer
>tannins and flavonoids from chestnut inner shell, shown to have antioxidant and deodorizing
activities

6. Health factor citrus flavonoids; with bioflavonoid (Vit P) synergistic activity

- citrus flavonoids with bioflavonoid (Vit P) synergistic activity

COMPOUNDS WITH COMPLEX SUBSTITUENTS WITH DEEPER COLORS

>Napthoquinone and anthraquinone deep purple to black
>Xanthone yellow
Ex. Xanthone mangiferine glucoside in mango
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FIRST LONG EXAM
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