S. MIZRAHI and M.

KAREL
Department of Nutrition & Food Science
Massachusetts Institute of Technology, Cambridge, MA 02139
ACCELERATED STABILITY TESTS OF MOISTURE-SENSITIVE PRODUCTS
IN PERMEABLE PACKAGES BY PROGRAMMING RATE
OF MOISTURE CONTENT INCREASE
ABSTRACT
A method for accelerated stability tests was developed, which is appli-
cable to isothermal storage of moisture-sensitive dehydrated products
packaged in water-vapor-permeable containers. The method does not
require prior knowledge of the kinetic model of the effect of moisture
on rate of deterioration. The method can be applied to dehydrated
products when the index of deterioration [defined as J - (dc/cn) where
c is reactant concentration, and n is the order of reaction] is dependent
only on moisture content, which changes continuously during storage.
The accelerated tests are based on monitoring quality changes in the
product, which undergoes rapid deterioration because of a high, albeit
controlled, rate of moisture gain. Theoretical considerations based on
I kinetic theories indicate that the rate at which the index of deteriora-
tion changes at any given moisture content is inversely proportional to
the rate of moisture gain. This fact provides a basis for the mathe-
I
matical procedures of extrapolation and prediction of storage stability
of the same product at the same temperature under any given rate of
moisture gain. The method successfully predicted the loss of ascorbic
acid in stored tomato powder and the extent of browning in dehy-
drated cabbage.
INTRODUCTION
DURING the past several years, techniques have been devel-
oped to predict the shelf life of packaged stored foods on the
basis of laboratory tests on kinetics of deterioration and on
mass transfer properties of packaging materials. Reviews of
this subject have been published recently by Karel (1973,
1975), Herrmann (1974), and Labuza (1972, 1973). The re-
lated subject of prediction of nutrient retention during proc-
essing was treated in papers by Lund (1973), Mulley et al.
(1975), and Teixera et al. (1969). The influence of packaging
on nutrient retention in food was reviewed by Karel and
Heidelbaugh (1975). Prediction of the shelf life of oxidation-
sensitive foods was the subject of papers by Singh and Held-
man (1976), Singh et al. (1975), Quast et al. (1972), Quast
and Karel (1972, 1973), Karel (1974), and Herlitze et al.
(1973).
A major shortcoming of these techniques for predicting
storage stability is the relatively long time required to evaluate
the kinetic parameters of deterioration. The experimental
design is usually based on a balanced matrix of samples in
which only one variable is changed at a time, usually over a
wide range of values. The mass transfer constants for packag-
ing materials, on the other hand, can be rapidly evaluated by
currently available methods (Karel, 1973, 1975; Karel et al.,
1971).
One of the major problems, therefore, is to develop effec-
I
tive accelerated test procedures. Mizrahi et al. (1970a) success-
fully conducted accelerated tests for nonenzymatic browning
in dehydrated cabbage by simultaneously increasing the mois-
ture content and the temperature of the samples. However,
these tests were made possible by prior knowledge of the
kinetic model for that specific system (Mizrahi et al., 1970b).
The knowledge of the kinetic model was essential in extrapo-
lating the obtained data to conditions of low reaction rates
(low temperature and moisture contents).
Temperature and concentration are variables for which
kinetic models are often available. The Arrhenius equation is
usually applicable as the model for temperature dependence,
and concentration dependence is commonly expressed in
order of reaction terms.
In contrast to the well-established relationships among rates
of reaction, temperature, and concentration, there are no
generally accepted models for the other variables. Some
models have been suggested in the literature - e.g., for the
effects of moisture content (Ross, 1948; Legault et al., 1951;
Mizrahi et al., 1970b; Quast and Karel, 1972) and oxygen
(Karel, 1960; Quast and Karel, 1972). However, all the pro-
posed models have a more or less limited validity, and their
usefulness in accelerated tests in cases other than the specific
systems from which they were derived has yet to be estab-
lished. Therefore, accelerated test development usually has to
be undertaken without a kinetic model. In such cases, it is
desirable to be able to obtain the necessary kinetic data with-
out having to establish a kinetic model. To our knowledge,
however, such a no model approach has never been re-
ported.
This work was undertaken to develop and establish the
feasibility of a no model accelerated test technique for de-
hydrated products undergoing deterioration because of
moisture-sensitive reactions.
EXPERIMENTAL
NONENZYMATIC BROWNING in dehydrated cabbage and loss of
ascorbic acid in tomato powder were the two moisture-sensitive reac-
tions used to test the feasibility of the no-model approach to acceler-
ated testing.
Data for the nonenzymatic browning reaction at 37C were taken
from Mizrahi et al. (1970a). The packaging materials used in that study
were PE (2.5 mil thick polyethylene) and SP48 (Scotchpak, Minnesota
Mining and Manufacturing Co.). Storage simulation, where indicated,
was based on the following equations:
Sorption isotherm:
m - 0.579
a = m + 9.583
(1)
where a = water activity and m = moisture content (g H, O/lOOg solids).
Kinetic equation:
A!!! = 135.5a3.05
dt
where dBr/dt is the rate of browning in Klett units/day.
The simulation of storage for packaged dehydrated cabbage was
carried out on a Wang Model 380 programmable calculator according to
the procedure outlined by Mizrahi et al. (1970b).
The data on retention of ascorbic acid in tomato powder were ob-
tained experimentally. Tomato powder, irjtially containing 142 mg
958 - JOURNAL OF FOOD SCIENCE - Vol. 42, No. 4 (1977)
11
STABILITY TESTS OF WATER-SENSITIVE FOODS.. .
ascorbic acid/lOOg solids, was produced by freeze drying commercially
frozen, fourfold-concentrated tomato juice (Vitality brand, Lykes
Pasco Co., Dade City, Fla). Samples of lo-12g of tomato powder were
packaged in 4- x lo-cm pouches of different flexible films of various
thicknesses - PEl, PE2, PE3 and PE4 (low-density polyethylene, 1, 2,
3 and 4 mil thick, respectively), HDPE/PiB 3 (blend of highdensity
polyethylene and polyisobutylene 3 mil thick) - supplied by Conti-
nental Can Co.
The packaged samples were placed in desiccators over a saturated salt
solution at the desired water activity (NaCl when a = 0.75, and
Na, Cr, 0, when a = 0.53). The desiccators were kept in the dark in a
constant-temperature cabinet maintained at 37.0 + 05C. Samples were
withdrawn for weighing and ascorbic acid determination according to a
predetermined schedule.
Absolute values for the water vapor permeability constant of the
packaging film, where indicated, where determined gravimetrically on
packages filled with desiccant. Permeability ratios, on the other hand,
were determined by in in-storage weight changes of each group of sam-
ples stored at the same external water activity. The calculations were
based on the fact that, at a constant external water activity and solids
weight, the time required to reach any given moisture content is in-
versely proportional to the permeability. Results are expressed in terms
of either permeability, kH, 0, or rate of moisture gain, K, as defined by
the following equation:
$=K(a,--al)=100
kHioio A (ae - ai)
s
where ae = external water activity; ai = in-package water activity at
equilibrium with moisture content of package contents; kH,o = per-
meability to water vapor; P, = water vapor pressure at storage tempera-
ture; A = package area; X = thickness of packaging material; W, =
weight of solids per package; and dm/dt = rate of moisture gain (g
H, O/lOOg solids/day).
The amount of ascorbic acid was determined by potentiometric
titration with 2,6dichloroindophenol using a platinum electrode aged
in water (Spaeth et al., 1962). Samples were prepared for titration by
suspending 2.5g of tomato powder in a 35-ml HPO,HOAc solution
(AOAC, 1975). After the suspension was stirred for 30 min, the sus-
pended particles were precipitated by 30 min of centrifugation at 5000
rpm (%3000 x G) in a Servall rotor-type SS-1 centrifuge. A 25-ml
aliquot of the supernatant was taken for titration. At least two repli-
cates of each sample were analyzed.
Curve fitting was carried out using a polynomial curve-fitting BMD
PSR Program (Dixon, 1975) on an IBM 360 system at the MIT Infor-
mation Processing Center.
RESULTS & DISCUSSION
THE NO-MODEL accelerated tests were formulated and tested
on the system shown schematically in Figure 1. This system
has the following properties: (1) the moisture content of the
product is increased continuously with time because of storage
conditions in which a, is always larger than ai; (2) the deterio-
ration rate in the dehydrated product is known to depend only
on the momentary moisture content; and (3) the order of
reaction (n) is known. Under these circumstances the kinetic
equation reads:
dc
--$ = cn f(m)
where c = reactant concentration, and t = time.
For sake of simplicity, an index of deterioration (D) was
defined as:
D -Do =cj-, (-2)
(5)
where D, is a reference value at cc (which in many cases could
be arbitrarily assigned as zero).
For the two common cases of zero and first order of reac-
tion, D assumes the values of D - D, = co - c and D - D, =
ln(c,/c), respectively. It should be noted that, by definition, D
T = CONSTANT
EXTERNAL WATER ACTIVITY = a,
lr I
INTERNAL WATER ACTIVITY = ai
/
Hd U I&ERI~RAT~~N RATE = f(rfJ j
Li u
Fig. l-Schematic representation of deterioration of moisture-
sensitive, packaged dehydrated foods.
always assumes a positive value, which can be calculated by
integrating Eq (4), i.e.,
D - Do =I0 f (m)dt
(6)
When moisture is introduced into the food system at a
constant rate (b), i.e.:
m=m, +bt (7)
where m, = initial moisture content, then dt can be replaced
by dm/b in Eq (6) to yield:
D - Do =rlrno f (m)dm =$[f(m) - f(mo)j
(8)
where f(m) = If (m)dm.
For cases in which b, m,, and m are kept constant, the
change in the index of deterioration is inversely proportional
to b. Therefore, when moisture is introduced into two samples
at two different but constant rates (br and ba), and when
both samples have the same m, and m, the index of deteriora-
tion ratio will be:
@-Do)2 =%
CD --Do)1 bz
Therefore, an experiment in which accelerated deteriora-
tion is produced by rapid moisture gain can be used to predict
the extent of deterioration in samples in which moisture is
gamed very slowly.
The simplest and most convenient way to control aceu-
rately the rate of moisture introduction is to use a controlled-
permeability package. In such cases, the rate of moisture gain
is no longer constant, although the moisture gain curve can be
broken into constant rate periods, at least for relatively small
increments of moisture gain. In the case of an infinitesimal
Vol. 42, No. 4 (1977) - JOURNAL OF FOOD SCIENCE - 959
increment of m (dm), the following equation can be applied If the changes in the index of deterioration are known for a
reference sample (1) in the following forms:
(D-D,)1 =F,(m)
and
(12)
dD2 /dm _ dt2 /dm
dDi/dm dt, /dm
(10)
The value of dt/dm-can be evaluated by Eq (3), and the
sorption isotherm [a = f(m)] can be incorporated to give:
dt 1
-=
dm K[a, - flm)]
(11)
0 100 200 300 400 500 600 7
MOISTURE GAIN (me.1
Fig. P-Ascorbic acid retention as a function of moisture gain in pack-
ages of differing warer vapor permeability (ho-a, sroragel.
2
1
01 I 1
0 100 200 300 400 500 600
MOISTURE GAIN (mg.1
Fig. 3-Ascorbic acid retention as a function of moisture gain, trans-
formed to a common reference rare of moisture gain (permeability
ratio of 1.81 fiso-a, storage).
dD1 _
--
dm
FI (ml
then the index of deterioration of the second sample (D2) can
be calculated as follows:
m
KI [ae 1 - T(m)1
Oh - Do)2 = lm, Flh) K2 la,2 _ qm)l
dm (13)
In a specific case in which external water activity is the
same in both samples, Eq (13) reduces to:
(D - D,)z =gmJm, Fi(m)dm =2 (D - D,)i (14)
As mentioned above, the validity of the no-model approach
outlined here was tested on two moisture-sensitive deteriora-
tion reactions, i.e., nonenzymatic browning of dehydrated
cabbage, which was found to be a zero-order reaction (Mizrahi
et al., 1970b), and.loss of ascorbic acid in tomato powder,
which is a first-order reaction (Riemer, 1976).
Knowledge of the order of reaction is essential for deter-
mining the mathematical form of the index of deterioration
and, therefore, should be established prior to any further cal-
culation.
The test cases can be divided into two groups: (1) acceler-
ated tests of reference samples and actual storage tests con-
ducted at the same external water activity (iso-a, tests); and
(2) reference and actual storage tests conducted at different
external water activities (hetero-a, tests).
Iso-a, tests
Figure 2 shows the results of the tests for ascorbic acid
retention in tomato powder; each curve represents a different
type of packaging film. At a given moisture gain, the extent of
deterioration is lower for the more permeable film, since less
time is required to reach that value of moisture gain. Never-
theless, these data, which were obtained in a relatively short
time, can be used to predict the extent of deterioration in a
less permeable film. This fact is demonstrated in Figure 3 by
the narrow distribution of the transformed data along a single
curve. This transformation of the original data (Fig. 2) has
been carried out by using Eq (14) for first order reaction and
the K ratio data in Table 1 (an example of calculation is shown
below). Therefore, under an iso-a, condition, data obtained
for any film can be used to predict the extent of deterioration
for the others.
In the following example, an actual fractional retention of
ascorbic acid of 0.86, which was found for PEI at a moisture
gain of 328 mg, is transformed, on an equal moisture gain
basis, to the K value of PE3, i.e., as if the sample were pack-
aged in PE3 and stored at the same a, until the same moisture
gain was reached. The predicted retention (using data in Table
1) is:
ln($2 =(D -Do)2 =yX ln& = 0.642
($)* = 0.53
960 - JOURNAL OF FOOD SCIENCE - Vol. 42, No. 4 (1977)
STABILITY TESTS OF WATER-SENSITIVE FOODS. . .
x
1.37 (t =14dayr)
o SP4S .224 (1=75dayr)
a, = .61
T = 37oc
I 2 3 4 5 6 7
MOISTURE GAIN (g. water/lOOg. solids)
Fig. I-Exrenr of nonenzymatic browning in dehydrated cabbage,
corrected for permeability of the film (SP48), as a funcrion of mois-
ture (iso-a, storage), where r is the time required to achieve the
maxi mum moisture gain shown.
A similar procedure was used to generate the results shown
in Figure 4. Data for browning of dehydrated cabbage pack-
aged in PE were used for prediction of browning in samples
packaged in SP48. Here again Eq ( 14) was used, but, in this
case, for a zero order reaction. If, for example, the sample
packaged in PE has a browning of Br = 71 Klett units (initial
browning, Br, = 44) at moisture gain of 5g H2 O/log solids,
then when packaged in SP48 at the same a, and moisture gain:
CD - Do)2 = (Br - BrO)* = s(71 - 44) = 165.
Br2 = 165 + 44 = 209 Klett units.
Therefore, it could be established that quality changes in a
packaged dehydrated product can be calculated conveniently
from accelerated tests in which deterioration at a high rate of
moisture gain is measured at iso-a, condition. The data in
Table 2 show another example where the loss of ascorbic acid
in tomato powder packaged in 1 mil thick polyethylene can be
used successfully to predict retention in much less permeable
films (PE4 or HDPE/PiB3). The time required to carry out the
accelerated test, however, is only about one-seventh of the
I 1 I
50 too 150
TIME (days)
Fig. B-Comparison of predicred and actual increases in browning of
dehydrated cabbage under simulated storage conditions (hereros, sror-
age).
actual storage time. This ratio could be improved further
either by using more permeable films in accelerated tests or by
increasing the ratio of package area to solids weight. The limit
to acceleration is defined by the sensitivity and accuracy of
the analytical methods used to determine the changes in the
quality of the product.
Hetero-a, tests
The data in Figure 5 show how well an accelerated test
utilizing a high rate of moisture gain (uptake, K = 2 at a, =
Table l-Rare of moisture gain of tomato powder packaged in dif-
ferent films, expressed as a ratio to bar of the PE3 sample
Film type
PEI
PE2
PE3
HDPE-PiB3
Ka ratio
(in ref to PE3)
4.26
1.96
1 .oo
0.55
e Determined by in-storage weight changes
Table 2-Comparison of predicted values and experimental tests of retention of ascorbic acid in tomato powder
Packaging Moisture gain
film type (g H,O/.lOOg solids)
Ascorbic acid retention
(%I
Predicteda Experimental
Required experimental
time (days1
Prediction Experimental
method method
PE 4 mil 2.25 70 f 3 69 2 1 2.15 14
PE 4 mil 2.52 63 f 3 63 +_ 1 2.85 19
HDPE/PiB 3 mil 2.25 70 f 3 70 f 1 2.15 14
a Based On data obtained for 1 mil thick PE film.
Vol. 42, No. 4 (1977) -JOURNAL OF FOOD SCIENCE -
961
0.75) predicted the extent of nonenzymatic browning in pack-
aged cabbage stored at a relatively low rate of moisture gain (K
= 0.2 at a, = 0.5). The data in Figure 5 are based on simulation
of storage of the dehydrated cabbage and on the procedure
.
mi
= mj., +Am
At, =
2Am 2Am
K, Da., -(a, +aj-,)l
At, =
I
K2 [2a.*- (aj +ai-,jl
I
AD, = F, (ml ) - F,(mj-,I OR AD, =[f(mi)-fhi-ljl 9
Fig. 6-Flow sheet for calculation of the deterioration index, D, for a
sample in any given package-a, combination from data obtained for the
same sample and temperature under known conditions.
- 0.1 -0.2
-013
INDEX OF DETERIORATION , D
outlined in Figure 6; the data are from Mizrahi et al. (1970a),
as indicated in the Experimental Procedures section. The cal-
culated values in Figure 5 are based on data that were obtained
in 24 days, whereas the actual storage time span was about
180 days. Therefore, the acceleration factor (ratio between
storage and accelerated test times) is 7.5 in this case.
The use of average values in the simulation procedure is the
reason for the apparent perfect prediction, since the experi-
mental errors are ignored. As stated before, however, experi-
mental errors are expected to be the main obstacle that must
be overcome when a high acceleration factor and a high level
of accuracy of the predicted values are desired.
In the storage simulation procedure, moisture gain curves
were generated by the mass transfer equation (Eq 3) plus the
sorption isotherm. However, in many practical cases, like that
of tomato powder (Fig. 7), it seems more convenient to deter-
mine the moisture gain directly by monitoring changes in
package weight. In such cases, a relatively simple graphic solu-
tion can be worked out on the basis of Eq (lo), since the
curves can be divided into a fairly small number of constant-
rate periods (Fig. 7). The index of deterioration for PE4 at a
given moisture gain (M,) can be calculated from a given curve
of Di vs m by the following expression (initial D is assigned a
value of zero):
D
t21
2x=D11- +@12 -Dll)
t22 - t21
t11
t
12 --tll
(15)
+(Dis -Dia:;: 1;;; +(Dl, -D&:2x --23
lx -t13
where Di, refers to the D of the ith sample at the end of the
nth constant rate period; and tin refers to t for the irh sample
at the end of the nth constant rate period.
Thus, e.g., for moisture gain (M,) equal to 1.8, the value of
Dax is 0.7589, which is equivalent to 47% of ascorbic acid
retention.
Figure 8 shows that the curve calculated by the graphic
method from data presented in Figure 7 accurately predicts
the actual ascorbic acid retention in actual storage. The curve
could be easily transformed to a time basis by using the mois-
ture gain curve in Figure 7 (for PE4).
Fig. 7-Representation of a graphic method for predicting ascorbic acid retention in tomato powder.
a@ -
film
-
.53 PE4
.75 PE3
I
IO 20 30
T I ME (days)
962 - JOURNAL OF FOOD SCIENCE - Vol. 42, No. 4 (1977)
I 11
STABILITY TESTS OF WATER-SENSITIVE FOODS. . .
In conclusion, an isothermal accelerated test procedure was
developed and found to be useful for predicting the extent of
deterioration of moisture-sensitive products packaged in water-
vapor-permeable films. The acceleration of the deterioration
process in the reference sample was achieved by subjecting the
packaged sample to a high rate of moisture uptake by using
films of a high-permeability constant, or increasing the ratio
package area to solids weight. The data from one reference
sample can be extrapolated to predict the extent of deteriora-
tion in the same product for any given package-external water
activity combination (as long as a, > ai). This calculation is
carried out by a mathematical procedure based on the fact
that the rate of change in the deterioration index is inversely
proportional to the rate of moisture gain.
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19 37oc
- Calculated valu18 (from PE 3 II. = ,751 (t=9,3doys)
. Actualdata (at PE4 a. .53) (t=26days)
0 I 2 3
MOISTURE GAIN (g. water/ 1009. solids)
Fig, 8-Comparison of predicted retention of ascorbic acid and ex-
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MS received S/21/76: revised 10/28/76: accepted 10/31/76.
Presented at the 36th Annual Meeting of the Institute of Food
Technologists, Anaheim, CA, June 6-9.1976.
This work was supported in part by Contract No. DAAK03-75-G
0038 with the U.S. Army Natick Laboratories. Dr. William L. Porter of
the U.S. Army Natick Research and Development Command was the
Project Officer, and the authors appreciated his discussion of this work.
This paper represents the position of the authors and not necessarily
that of the Department of the Army.
Vol. 42, No. 4 (1977) - JOURNAL OF FOOD SCIENCE - 963
I