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Effect of Water Activity on the Stability to

Oxidation of SprayDried Encapsulated
Orange Peel Oil Using Mesquite Gum (Prosopis
Juliflora) as Wall Material
ARTICLE in JOURNAL OF FOOD SCIENCE DECEMBER 2001
Impact Factor: 1.79 DOI: 10.1111/j.1365-2621.2002.tb11385.x

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Food Engineering and Physical Properties

Effect of Water Activity on the Stability to

Oxidation of Spray-Dried Encapsulated
Orange Peel Oil Using Mesquite Gum
(Prosopis Juliflora) as Wall Material
C.I. BERISTAIN, E. AZUARA, AND E.J. VERNON-CARTER

ABSTRACT: Mesquite gum solutions (30% w/v) were used to emulsify orange peel oil in an oil-gum solids ratio of
0.25. Emulsions were spray dried in laboratory scale equipment. The powders were stored in aw (s) from 0.108 to
0.743 at 35 8 C. Quantitative analysis of limonene oxide indicated that the sample at 0.628 showed a very good
stability against oxidation after thirty d, without caking and stickiness occurring. At this water activity the system
was within rubbery state, and the moisture content corresponded to that of the minimum integral entropy.
Keywords: mesquite gum, microencapsulation, minimum entropy, water activity, and glass transition

Introduction

Food Engineering and Physical Properties

UMEROUS MATERIALS ARE COMMERCIALLY AVAILABLE TO EN-

capsulate flavor using the spray drying technique. The

most widely used encapsulating agents are gum Arabic and
modified or hydrolyzed starches. Mesquite gum (Prosopis
juliflora) has been reported as having the ability to encapsulate orange peel oil (80.5% of the starting oil) but to a lesser
extent than gum Arabic (93.5% of the starting oil) (Beristain
and Vernon-Carter 1994). Goycoolea and others (1998) found
that the orange peel oil encapsulating capacity of gum Arabic
was slightly higher than that of a native and low-tannin mesquite gum (Prosopis spp.), when used in combination with
modified cornstarch carrier matrices. A blend of 60:40% wt
of gum Arabic to mesquite gum was able to encapsulate the
same amount of orange peel oil than pure gum Arabic
(Beristain and Vernon-Carter 1995), whereas a 3:2 ratio of
maltodextrin 10DE to mesquite gum retained 84.6% of the
starting orange peel oil, providing a better encapsulating capacity than either polysaccharide on its own (Beristain and
others 1999). However, mesquite gum is not yet commercially available. In most studies, temperature has been considered as the main parameter affecting the stability of encapsulated orange peel oil, whereas aw effects have been
neglected (Risch and Reineccius 1988; Chang and others
1988). One exception is the report by Anker and Reineccius
(1988), who monitored the formation of limonene oxides in
encapsulated spray dried orange peel oil stabilized in the aw
range of 0.001 to 0.536, finding that the slowest rates of formation of limonene oxide occurred at water activities of
0.536.
The glass transition has been proposed to accelerate diffusion-controlled reactions, such as oxidation, enzymatic reactions, and nonenzymatic browning (Slade and Levine
1991). These reactions in food systems are related to changes
in water content, water activity, temperature, pH, reactant
concentration, and structure (Bell 1996).
Diffusion in glassy systems is claimed to be virtually nonexistent. It is believed that low mobility in the glassy state due
to high viscosity makes chemical reactions improbable (Bell
and Hageman 1994). However, oxidation is dependent on
206

JOURNAL OF FOOD SCIENCEVol. 67, Nr. 1, 2002

proper matrix formation as well as exclusion of oxygen from

the matrix (Bell and Hageman 1995). The physical state of
the encapsulation matrix and its moisture absorption is critical in stopping oxidation development during storage (Shimada and others 1991). An amorphous glass entraps flavor
compounds and protects them from oxidation due to slow
diffusion (Onwulata and Holsinger 1995). Ross (1995)
claimed that flavor compounds that are susceptible to oxidation become protected from oxygen due to decreased diffusion of gases through the glassy structure below Tg. On the
other hand, Ma and others (1992) found that oxidation of oil
within a maltodextrin M 100 encapsulating matrix occurred
within the wall materials glassy state. Roozen and others
(1991) showed that the rotational mobility of tempol increased within the glass transition temperature of the system. Glassy state was not rate limiting. Bell and Hageman
(1994) found that the rate of aspartame degradation depended upon water activity rather than upon the state of the system, and that mobility, as dictated by the glass transition
temperature of the system, was not rate limiting. Andersen
and others (2000) followed the permeation of oxygen
through a glassy matrix encapsulating oil, finding that oxygen
permeation increased with temperature and was the rate-determining step.
Stability is greatly influenced by the moisture sorption
characteristics of the product. The thermodynamics of water
sorption in dried foodstuffs has drawn interest because it
provides a more thorough interpretation of the sorption isotherm phenomenon and helps to understand better the
sorption mechanism. There exists a controversial issue of
whether moisture equilibration represents a true state of
thermodynamic equilibrium, as certainly are all the spraydried materials. Although true equilibrium in thermodynamic terms is not achieved, the pseudo equilibrium states attained may be considered stable for practical time frames.
Thermodynamic parameters have been used to study the
stability of foods. Le Maguer (1985) presented the basic
framework for the use of solution thermodynamics and discussed its application to starch water systems. Rizvi (1986)
discussed general principles associated with hystereses and
2002 Institute of Food Technologists

Stability to oxidation of encapsulated orange peel oil . . .

how these affect thermodynamic calculations. Braibanti
(1990) calculated the chemical potential of binding water
molecules to reveal the specific features of adsorbing surfaces for cod, bee, pectin, and microcrystalline cellulose.
Beristain and others (1994, 1996) used Othmers equation to
calculate both differential and integral thermodynamic parameters to study stability of coffee beans, and applied the
enthalpy-entropy compensation for water adsorption of
starchy foods and dried fruits, finding that adsorption processes in microstructures with low moisture content were
entropy controlled and modified by temperature.
The purpose of this study was to investigate the relative
degree of protection against oxidation provided by mesquite
gum to orange peel oil encapsulated via spray drying stored
at several water activity conditions.

by the sample alone, the filter paper was allowed to equilibrate over the salts solution. Samples were placed in separate desiccators containing saturated salt slurries in the
range of water activity from 0.11 to 0.85 using the aw reported by Labuza (1985). The samples, were held at 25, 35, and
45 8C until equilibrium was reached. Values of water activity
were generated using equations reported in the same paper.
Equilibrium was assumed when the difference between 2
consecutive weightings was less than 1 mg/g of solids. The
time to reach the equilibrium varied from 8 to 14 d. Moisture
content of the humidified systems was determined by difference in weight after drying in a vacuum oven at 60 8C in the
presence of magnesium perchlorate desiccant.
The Guggenheim-Anderson-De Boer (GAB) equation was
used in modeling water sorption (Weisser 1985):

Materials and Methods

Mesquite gum, collected in the form of tears in the Mexican state of San Luis Potosi, was pulverized in a hammer mill
(Glen Mills Inc., Clifton, N.J., U.S.A.). In order to stop enzymatic activity, the powder was dissolved in water at 77 8C for
1 h in a stainless jacketed vessel (Polinox, Mexico, D.F., Mexico) with a propeller-type agitator, filtered with a high-flow
supercel in a Shriver filter press (EIMCO Process Equipment,
Houston, Texas, U.S.A.), and dried in a Bowen BLSA pilot
plant spray-drier (Somerville, N.J., U.S.A.) with an inlet air
temperature of 175 5 8C and an outlet air temperature of
87 5 8C.
Single strength orange peel oil was obtained from Derivados Veracruzanos, S.A. de C.V. (Xalapa, Ver., Mexico). All water
used in the experiments was double distilled and deionized.

Emulsion formation and spray drying

An aqueous mesquite gum solution was prepared from
spray-dried gum with a concentration of 30% (w/v). The solution was heated and held at 80 8C with constant stirring for 30
min, then allowed to cool, covered, and left to stand overnight
at room temperature. Emulsions were prepared by adding
drop wise and dispersing the requisite amount of orange peel
oil into the aqueous mesquite gum solution in order to obtain
a ratio of 25 g of oil/100 g gum solids, using a Silverson L4R
Heavy Duty Laboratory Mixer Emulsifier (Silverson Machines,
Ltd., Chesam., Bucks., U.K.) at a speed setting of 5. The resulting crude emulsions were then homogenized by increasing
the speed setting in the Silverson to 10 until a mean volumetric
globule size smaller than 2.5 mm was obtained, as measured
by a Malvern Droplet and Particle Size Analyzer series 2600
(Malvern Instruments, Malvern, Worcs., U.K.). Emulsion samples (about 400 g) were spray dried in a Bchi Mini Spray Dryer model 190 (Bchi Laboratoriums Technik AG, Flawil, Switzerland), using an inlet temperature of 200 5 8C and an
outlet air temperature of 110 5 8C.

where aw is water activity; M is water content of the sample

on dry basis; M o is the monolayer water content; C is the
Guggenheim constant = Cexp (hm-hn)/RT; hm is the heat of
sorption of the first layer, hn is the heat of sorption of the
multilayer, K is the constant correcting properties of multilayer molecules with respect to bulk liquid = k exp (h1-hn)/
RT; h1 is the pure water heat condensation; T is the absolute
temperature; and R is the gas constant. The parameter values
of GAB equation (Mo,C,and K) were obtained by nonlinear
regression analysis.
Goodness of fit was evaluated using the average of the relative percentage difference between the experimental and
predicted values of the moisture content or mean relative
deviation modulus (P) defined by the following equation (Lomauro and others 1985):

(2)
where Mi is the moisture content at observation i; MPi is the
predicted moisture content at that observations; and n is the
number of observations.

Thermodynamic properties

The isosteric heat of sorption is a differential molar quantity derived from the temperature dependence of the isotherm, and it represents the energies for water molecules
binding at a particular hydration level, in contrast to the integral heat, which is the average energy of all molecules already bound at that level (Schneider 1981). The respective
differential and integral entropies are obtained from their
differential and integral heats, respectively. The usual entropy discussed qualitatively or quantitatively (statistical mechanics) in terms of order-disorder of the adsorbed moleVapor sorption isotherms
cules is the integral entropy and not the differential entropy
Samples of the spray-dried encapsulated orange peel oil (Hill and others 1951, Rizvi and Benado 1984).
were placed in desiccators containing P2O5 for 3 wk at room
Differential properties. Changes in differential enthalpy at
temperature. The moisture sorption data were obtained us- the water-solid interface at different stages of the adsorption
ing the gravimetric method described by Lang and others process were determined using Othmers equation (1940):
(1981). Two to 3 grams of samples were weighed in triplicate
into standard weighing dishes with a circular section on the
bottom, where a quantitative filter paper Whatman No. 1 was
(3)
used to support the sample and at the same time allow transmission of moisture. In order to obtain a true moisture gain
Vol. 67, Nr. 1, 2002JOURNAL OF FOOD SCIENCE

207

Food Engineering and Physical Properties

(1)

Raw materials

Stability to oxidation of encapsulated orange peel oil . . .

where Pv is vapor pressure of water in the food; Pv0 is vapor
pressure of pure water at the same temperature; Hv( T) is
isosteric heat for water adsorption; Hv0(T) is heat of condensation of pure water; M is moisture; and C is adsorption constant.
A plot of lnPv against lnPv0 gives a straight line if the ratio
Hv(T)/Hv0(T) is maintained constant in the range of temperatures studied.
The net isosteric heat of adsorption or differential enthalpy is defined by Eq. 4.

(4)
Calculating Hv(T)/Hv0(T) with Eq. 3 and substituting into
Eq. 4, it is possible to estimate the net isosteric heat of adsorption at different temperatures using steam tables.
With values obtained for enthalpy changes, the variation in
the molar differential entropy (DSdif)T may be estimated using
Eq. 5:
(5)

Food Engineering and Physical Properties

where S1 = (S/N1)TiP is molar differential entropy of water

adsorbed in the food; SL is molar entropy of pure water in
equilibrium with the vapor; S is total entropy of water adsorbed in the food; N1 is number of moles of water adsorbed
in the food; R is universal gas constant; aw is water activity;
and T is temperature (K).
Integral properties. Molar integral enthalpy is calculated
using an expression similar to that for differential enthalpy,
maintaining diffusion pressure constant:

(6)

Table 1Estimated parameters of the GAB equation for

orange peel oil microcapsules
Temperature 8 C
Parameter
C
K
Mo (% dry basis)
P%

25

35

45

12.04
0.918
5.74
4.96

15.33
0.933
5.34
3.41

18.9
1.02
4.45
4.75

Storage stability
Thirty-gram samples were placed in desiccators containing saturated solutions of LiCl, KC 2H3O 2, MgCl 2, K2CO 3,
Mg(NO3)2, NaNO2, NaCl for 30 d. At 35 8C the water activities
of the desiccants were 0.108, 0.215, 0.318, 0.436, 0.515, 0.628,
and 0.743 respectively. Portions of the samples were withdrawn every 5 d for gas chromatographic analysis. The samples were put into the desiccants immediately after they
were spray-dried, and this was taken as the zero point time.

Gas chromatographic analysis

Samples were prepared for gas chromatography as described by Anker and Reineccius (1988). In short, 0.15 g powder were dispersed in 0.85 g distilled water in a screw cap
vial, then mixed slowly with a vortex into 4 ml of acetone
containing 0.25 mg/ml of 2-octanone as internal standard.
The resultant supernatant was injected without further preparation into a Hewlett-Packard model 6890 gas chromatograph (GC) (Hewlett Packard Co., Wilmington, Del., U.S.A.).
The GC was equipped with flame ionization detector and
HP-5 column. The carrier gas was helium at a flow velocity
of 1 ml/min. The temperature of the column was programmed initially at 50 8C for 1 min and then increased at
rate of 8 8C/min to a final temperature of 200 8C. The
amount of limonene oxide was determined by the internal
standard method (Anandaraman 1984).

where Hvi( T) is integral molar heat of water adsorbed in Glass transition

food and F can be found by (Nunes and Rotstein 1991):
The glass transition temperature was determined by a differential calorimeter (DSC 2910, TA Instruments, New Castle,
Del., U.S.A.), equipped with a refrigerated cooling accessory.
(7) The Thermal Solutions Instrument Control and Universal
Analysis Software were used (TA Instruments, New Castle,
Del., U.S.A.). The samples (8 to 10 mg) were transferred to
aluminum pans, sealed hermetically, and weighed. The calo(8) rimeter was calibrated with indium (melting point
156.598 8C) and mercury (melting point -38.834 8C). Three
where F is diffusion pressure or surface potential of the replicates were carried out for each analysis. Samples were
food; ma is chemical potential of the adsorbent in the con- heating at 20 8C/min to a temperature that was 30 8C above
densed phase; map is chemical potential of the pure adsor- the estimated Tg from preliminary runs. Then it was cooled
bent; Wap is molecular weight of the adsorbent; Wv = molec- at 10 8C/min and reheated at 5 8C/min. Glass transition temular weight of water; F/a1 constant is similar to a process at perature was determined as the onset point of the step
change on the heat flow curve.
F constant.
When values for (DHint) are obtained, changes in molar
Results and Discussion
integral entropy can be calculated using Eq. 7:
HE GAB MODEL WAS FOUND TO AGREE VERY WELL WITH THE
experimental data. The parameters from fitting the GAB
model
were determined (Table 1). The mean relative devia(9)
tion modulus value was less than 5% at 25, 35, and 45 8C. Water activity of foods can be related to stability and the rates
where SS = S/N1 is integral entropy of water adsorbed in the of deteriorative reactions. It has also been assumed that the
food.
monolayer value is the critical water content where dehy-

208

JOURNAL OF FOOD SCIENCEVol. 67, Nr. 1, 2002

Stability to oxidation of encapsulated orange peel oil . . .

drated foods are most stable. Encapsulated orange peel oil
prepared with mesquite gum at 35 8C had a monolayer value
of 5.34 g water/100 g solids calculated by the GAB equation.
Differential and integral entropies changed with respect
to moisture content at 35 8C for the encapsulated orange
peel oil prepared with mesquite gum (Figure 1). The intersection of the curves is found at the minimum integral entropy, and it can be observed that, as the microcapsules adsorb
moisture, their integral entropy falls to a minimum. It is at
this point that maximum stability can be assumed, since water molecules are more ordered within the microencapsulating material (Nunes and Rotstein 1991). In our experiments,
the point of maximum stability against oil oxidation was
found at 12.8 g water/100 g soluble solids for microcapsules
prepared with mesquite gum, corresponding to a water activity close to 0.628. The point of minimum integral entropy
can be interpreted as the moisture content that corresponds
to the monolayer (Hill and others 1951; Nunes and Rotstein
1991). Minimum entropy values may be expected where
strong bonds between adsorbate and adsorbent occur
(Nunes and Rotstein 1991), and therefore, water is less available to participate in spoilage reactions.
The point of minimum integral entropy, and theoretically
of maximum stability, is different than the calculated values
for the GAB monolayer for the mesquite gum microcapsules
with 5.34 g water/100 g soluble solids (Figure 1).
Differential entropy had a minimum at 5.7 g water/100 g
dry solids that was similar to the GAB monolayer value. Nevertheless, this parameter does not mean order or disorder of
the total system. The differential entropy represents the algebraic sum of the integral entropy at a particular hydration level, plus the change of order or disorder after new water molecules were adsorbed by the system at the same hydration
level. If the values of moisture content corresponding to minimum integral entropy and minimum differential entropy are

Table 2Glass transition temperature (Tg) of encapsulated

orange peel oil with mesquite gum storage at 35 8 C as a
function of water activity (aw) and moisture content (M)

Figure 1Differential and integral entropy changes as a

function of moisture content for encapsulated orange oil
stored at 35 8 C

Figure 2Glass transition temperature of orange peel oil

microencapsulated in mesquite gum stored at 35 8 C as a
function of water activity

aw
0.215
0.318
0.436
0.515
0.628
0.743
0.821

Tg 8 (C)a
86.41 0.28
71.35 0.32
52.07 0.26
30.65 0.20
20.35 0.21
- 4.10 0.20
- 10.51 0.21

M (g water/100 g s.s.)
4.910
6.850
8.900
10.06
12.23
15.83
22.90

0.15
0.17
0.16
0.22
0.23
0.29
0.25

different, this particular hydration level at the minimum differential entropy cannot be considered as the maximum stability point, because not all available active sites have been occupied at that particular water content, and therefore it is
possible to obtain after this point lower differential changes
that provide a better ordering of the water molecules adsorbed on food. The water content at the minimum differential entropy in this case was similar to the GAB monolayer value because the maximum entropy change was provoked by
the water sorption on the most active sites. Monolayer value is
the saturation of polar groups corresponding to water adsorbed at the most active sites.
The temperature (Tg) at which microcapsules have their
glass transition depends strongly on the water content. The
Tg temperatures decreased with increasing water content as
can be observed in Table 2. Glass transition was difficult to
detect in mesquite gum at low moisture content < 4.91 g water/100 g of dry solids. The inability to find the glass transition at low moisture contents was consistent with previous
findings for complex food systems (Bell and Touma 1996).

Vol. 67, Nr. 1, 2002JOURNAL OF FOOD SCIENCE

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Food Engineering and Physical Properties

a Mean of 3 replicates 6 standard deviation

Stability to oxidation of encapsulated orange peel oil . . .

Food Engineering and Physical Properties

The Tg values were also plotted against water activity. This

plot showed a linear relationship between water activity and
glass transition temperature (Figure 2). Prediction of the Tg
with the linear relationship between Tg and aw allows a rapid
and fairly reliable method for locating the Tg of materials
stored at various conditions before experimental verification
of the Tg is obtained (Roos 1995).
This GAB monolayer value corresponds to a water activity
of 0.26 and a Tg = 79 8C, and this Tg is above the storage temperature 35 8C. Based on the properties associated with the
glassy state, where diffusion is claimed to be virtually nonexistent (Levine and Slade 1989), it would be expected that no
oxidation of the orange oil should occur at all. Nevertheless,
the rate of oxidation of the orange oil, contained within the
mesquite gum-encapsulating matrix, occurred rapidly within
the glassy state. Oxidation rates increased as the water activity of the microcapsules decreased from 0.436 to 0.108, but
were still within the glassy state (Figure 3). In contrast, the
microcapsules in the aw range of 0.515 and 0.628, within the
rubbery state, showed the greatest stability, when an accelerated rate of oxidation was expected based on the results
found for the microcapsules in the aw range from 0.436 to
0.108. The mesquite gum microcapsules exposed to a w of
0.743 showed the best stability against oxidation, but the data
from this water activity were not included in Figure 3 because microcapsules showed visual structural changes and
water gain by the powders led to a change in their flow properties as a result of caking and agglomeration.
A aw of 0.821 resulted in the complete destruction of the
microcapsules. At this humidity level, sufficient water was
drawn up by the microcapsules to form a paste-like mass.
These results seem to indicate that storing microcapsules
at aw(s) corresponding to the zone of minimum integral entropy provides them with the best stability against oxidation.
Levine and Slade (1989) claimed that low mobility in glassy
state makes chemical reactions improbable. However, the numerous mechanisms of chemical reactions make such a generalization unfounded (Bell and Hageman 1995). Oxidation is
dependent upon proper matrix formation as well as exclusion

of oxygen from the matrix. Glassy characteristics may delay

oxidation only if the oxygen is suitable to be entrapped by the
molecular structure of the glassy matrix. Andersen and others
(2000) showed that in oil encapsulating glassy food matrix,
some oil particles might oxidize rapidly and some more slowly, due to heterogeneity in the degree of encapsulation. The
rate of oxidation should be regarded as an average of all individual oxidation processes taking place in each of the oil particles. In the present work, probably the major determinant of
the microcapsules shelf life is the porosity of the dried matrix
to oxygen diffusion, independently of the supercooled liquid
state of the matrix. With values of aw (s) > 0.5, that were definitely above the calculated values for the GAB monolayer region, an increase in the rate of oxidation is expected. Likewise,
the protection and retention of volatiles is expected to occur
as long as the microcapsules wall structure remains intact.
The gel-like structure arising from the water, adsorbed by the
mesquite gum, acts as a shell opposed to oxygen diffusion into
the microcapsule core, inasmuch as water is adsorbed without initiating the wall dissolution process. However once the
capsule structure is damaged by water uptake, volatiles are increasingly exposed. Once the structure is destroyed, as in the
case of water activity at aw of 0.821, the capsules lose their volatiles content.

Conclusion

ESQUITE GUM PROVIDED A VERY GOOD PROTECTION

against oxidation to orange peel oil as long as microcapsules remained stored at the proper water activity. Oxidation processes occurred with different intensities at the various water activities. Microcapsules in the aw range of 0.515
and 0.628 showed the greatest stability. At water activity below 0.628, the structure of the capsules was not damaged; increasing water activity to between 0.743 and 0.821 led to
gradual dissolution of the walls. Results suggest that oxidation of orange peel oil occurred when the samples were in
glassy state, and oxidation was slower when they were rubbery. Based on the properties of the glassy state of the microcapsules, glassy systems were stable to physical changes
(agglomeration, caking), whereas rubbery systems were stable to oxidation and physical changes as long as the capsule
structure was intact.
The minimum integral entropy can be interpreted as the
water activity (0.628) at which the microcapsules have the
best stability to oxidation. The system at this water activity
was within the rubbery state, showing the best stability.

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Figure 3Effect of water activity on limonene oxidation of

encapsulated orange peel oil stored at 35 8 C
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The authors gratefully acknowledge financial aid by the Consejo Nacional de Ciencia y Tecnologia
de Mexico, who supported this study through the projects 25924-B and G33565-B.

Authors Beristain and Azuara are with the Instituto de Ciencias Bsicas,
Universidad Veracruzana, Apdo. Postal 575, Xalapa, Ver., Mxico. Author
Vernon-Carter is with the Universidad Autnoma MetropolitanaIztapalapa, San Rafael Atlixco #186, 09340 Mxico, D.F. Direct correspondence to author Beristain (E-mail: cberista@bugs.invest.uv.mx).

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