Blackwell Science, LtdOxford, UKJSSJournal of Sensory Studies0887-8250Copyright 2004 by Food & Nutrition Press, Inc., Trumbull, Connecticut.

196500509Original Article SHELF-LIFE ESTIMATION BY SURVIVAL ANALYSIS A.

GÁMBARO

ET AL.

SENSORY SHELF-LIFE ESTIMATION OF ALFAJOR BY SURVIVAL

ANALYSIS

ADRIANA GÁMBARO1, ANA GIMÉNEZ and PAULA VARELA

Sección Evaluación Sensorial, Facultad de Química

Universidad de la República
Avda. Gral. Flores 2124, C.P.11800
Montevideo, Uruguay

AND

LORENA GARITTA and GUILLERMO HOUGH

Departamento de Evaluación Sensorial de Alimentos

Instituto Superior Experimental de Tecnología Alimentaria
Nueve de Julio
Buenos Aires, Argentina

Received for Publication May 7, 2004

Accepted for Publication August 8, 2004

ABSTRACT

Survival analysis methodology was used to estimate the shelf life of

alfajor (a chocolate-coated individually wrapped cake) at 20 and 35C by using
results obtained from consumers when asked if they would accept or reject
samples with different storage times.
Sensory acceptability (measured by consumers), off-flavor (measured by
a trained panel) and moisture content were linearly related to time. These
correlations were used to estimate values at the shelf-life times calculated for
25 and 50% rejection probability.
Survival analysis provided the following shelf-life estimation: 74 days at
20C and 33 days at 35C for a 25% of rejection, 87 days at 20C and 39 days
at 35C for a 50% of rejection.
An alfajor stored at 20C having an acceptability value below 4.9 (1–9
hedonic scale) and off-flavor intensity above 5.3 (0–10 scale) would be
rejected by 25% of the consumers.
Chemical data were not good shelf-life predictors.

1
Corresponding author. TEL: +598-2-9245735; FAX: +598 2 9241906; EMAIL: agambaro@fq.edu.uy

Journal of Sensory Studies 19 (2004) 500–509. All Rights Reserved.

500 © Copyright 2004, Blackwell Publishing
 SHELF-LIFE ESTIMATION BY SURVIVAL ANALYSIS 501

INTRODUCTION

Alfajor is a chocolate-coated individually wrapped cake that is popular

in some Ibero-American countries such as Argentina and Uruguay. It is
defined by Uruguayan legislation as a product obtained by joining two or more
layers of cookies or cake with filling, usually dulce de leche (milk sweet),
covered or enrobed with various confectionary coatings, chocolate being the
most common (M.S.P. 1994).
In Uruguay, the alfajor market is about 25 units per inhabitant per year.
Seventy-three percent of the population is a frequent consumer of the product,
increasing to 90% when considering the population under 12 years old
(Equipos Retail 2001; Grupo Radar 2001). It is widely used as a snack at
school, work and leisure time. In spite of its wide and popular use, there
are no published sensory studies.
Alfajor belongs to the category of intermediate moisture food prod-
ucts, i.e., with moisture content below 20% and water activity (aw)
within the range 0.75–0.80 (information provided by the manufacturer). It
is made up of a wide range of ingredients, with flour confectionery in
contact with chocolate and sugar confectionery. The latter can complicate
shelf-life considerations when combined with flour-based products. Choc-
olate not only adds a unique flavor and texture but also encloses the
product in a fatty barrier that reduces the rate of moisture loss from the
cake and thereby assists in extending shelf life (Jones 2000). The alfajor
sold in Uruguayan supermarkets usually has a minimum shelf life of
2 months.
Alfajores are stored at room temperature, which is approximately 20C in
most situations. The product could be stored at higher temperatures, the
maximum being approximately 35C. Shelf-life calculations at different tem-
peratures allow estimating the activation energy to, therefore, be able to
estimate shelf life at other temperatures.
Survival analysis is a branch of statistics used extensively in clinical
studies, epidemiology, biology, sociology and reliability studies (Kleinbaum
1996; Klein and Moeschberger 1997; Meeker and Escobar 1998; Gómez et al.
2001; Gómez 2002). Recently, Hough et al. (2003) have introduced this meth-
odology to the study of food shelf life. Their key concept was to focus the
shelf-life hazard on the consumer rejecting the product, rather than on the
product deteriorating.
The objectives of the present work were to (1) study how stor-
age time affects consumer acceptability, trained sensory panel mea-
sures and chemical measures; and (2) determine the shelf life of the
alfajor stored at 20 and 35C based on consumer’s acceptance/rejection
of the product.
502 A. GÁMBARO ET AL.

MATERIALS AND METHODS

Samples
Alfajores used in all the experiments were provided by Durulte S.A.
(Montevideo, Uruguay). According to the information provided by the
manufacturer, the following ingredients were used in the products’
manufacture:

(a) Cookie: flour, sugar, hydrogenated vegetable oil, high fructose corn
syrup, eggs, cacao, ammonium bicarbonate (E-503i), sodium bicarbonate
(E-500ii), artificial vanilla flavor, soy lecithin (E-322), caramel (E-150)
and calcium propionate (E-282).
(b) Coating: sugar, hydrogenated vegetable oil, cacao, powdered skimmed
milk, lecithin (E-322), artificial vanilla flavor and sorbic acid (E-200).
(c) Filling (Milk sweet): whole milk, sugar, glucose, carrageenan (E-407),
sodium bicarbonate (E-500ii), artificial vanilla flavor and potassium sor-
bate (E-202).

Composition, provided by the manufacturer, was carbohydrates, 62.5%; fat,

17.5%; protein, 5.0%; ash, 2.0% and water, 13.0%. Alfajores were wrapped
individually in 20-m bioriented polypropylene with a metallic coating. Each
alfajor weighed 40 g.

Storage Temperatures and Sampling Times

To study the effect of storage temperature, shelf-life experiments were
conducted at 20 and 35 ± 0.5C. Storage times were selected according to
Gacula’s (1975) criterion, increasing the number of samples in the period in
which the product was likely to fail. The resulting sampling times at each
temperature were, at 20C: 0, 28, 46, 60, 65, 70, 75 and 80 days; and at 35C:
0, 10, 16, 25, 30, 35, 39 and 45 days. Samples were manufactured for each
of the sampling times and stored at the corresponding temperatures. As sam-
ples belonged to different batches, the trained sensory panel performed trian-
gular tests to ensure that there was no significant difference between a sample
of the previous batch (kept at 5C) and that of the following batches.
The study was carried out in two stages:

(a) Analysis of the samples with different storage time at 35C.

(b) Analysis of the samples with different storage time at 20C.

Previous microbiological analysis (aerobic mesophiles, coliforms, yeasts and

molds) showed that the samples were fit for consumption. The ethics commit-
 SHELF-LIFE ESTIMATION BY SURVIVAL ANALYSIS 503

tee of Universidad de la República concluded that all samples were adequate

for test on humans in the quantities to be served.

Sensory Analysis
The shelf-life panel consisted of eight assessors with a minimum of
18 months of experience in discrimination tests and in descriptive analysis of
bakery and confectionery products.
To identify those defects most likely to appear as a result of prolonged
storage, a preliminary test was performed, in which five samples with different
deterioration times were presented to assessors, who had to write down the
descriptors that made those samples different. By open discussion with the
panel leader, assessors agreed on the best descriptors to differentiate the stored
samples from the fresh sample: alfajor dryness and off-flavor. Off-flavor
encompassed flavors such as moldy, acid and/or rancid. During training,
assessors also referred to that descriptor as aged or altered. Once the descrip-
tors were selected, assessors were trained measuring samples with different
storage times, using 10-cm unstructured scales anchored with “nil” and
“high.”
The testing was carried out in a sensory laboratory equipped with indi-
vidual booths and white fluorescent daylight-type illumination. Two quarters
of an alfajor were served in plastic odorless trays at room temperature. Water
was used for rinsing between samples. For scoring, 10-cm unstructured scales
anchored with “nil” and “high” were used.

Acceptability Test
Consumers were recruited among students and teachers from Facultad
de Química, Montevideo, Uruguay. Each study, corresponding to 20 and 35C
storage temperatures, was carried out with 50 respondents who consumed
alfajores at least once a week. Approximately half of them were female and
half were male with ages ranging from 18 to 35 years. Consumers received
one-fourth of each sample (eight samples with different storage time in each
study) in random order. For each sample, they had to score the product using
a scale with nine boxes anchored on the left with “dislike very much,” in the
middle with “indifferent” and on the right with “like very much.” They also
answered the question, “Would you normally consume this product?” with a
yes or a no.

Chemical Determinations
Moisture and titrable acidity were measured by standard techniques
(AOAC 1995).
504 A. GÁMBARO ET AL.

Statistical Analysis
Analysis of variance (ANOVA) was performed on the trained sensory
panel data for each temperature, using storage time, assessor and their inter-
action as variation factors. On the consumer data, the interaction between
storage time and consumer as variation factors could not be calculated, as each
consumer measured each sample only once.
Survival analysis methodology was used to estimate the shelf life of
alfajores at 20 and 35C, using the results obtained from consumers when asked
if they would normally consume the samples with different storage times.
In food shelf-life studies, samples with different storage times are pre-
sented to consumers. Assume that a random variable T is defined as the storage
time on which the consumer rejects the sample. Thus, the survival function
S(t) can be defined as the probability of a consumer accepting a product
beyond time t, that is S(t) = P(T > t). Alternatively, the cumulative distribution
function F(t) = 1 - S(t), can be defined as the probability of a consumer
rejecting a product before time t, that is F(t) = P(T £ t).
Because of the discrete nature of the storage times, T will never be
observed exactly and thus the censored nature of the data (Hough et al. 2002).
Suppose that consumers are presented with samples stored at times a, b and
c. If a consumer rejects the sample at the first storage time observed, then
T £ a, and the data are left censored. If a consumer accepts the sample stored
at time a but rejects the sample stored at time b, then a < T £ b, and the data
are interval censored. Finally, if a consumer accepts all samples, then T > c,
and the data are right censored.
The likelihood function, which is used to estimate the survival function,
is the joint probability of the given observations of the n consumers (Klein
and Moeschberger 1997):
L = P S( ri ) P [1 - S(li )] P [ S(li ) - S( ri )] (1)
i ŒR i ŒL i ŒI

where R is the set of right-censored observations, L the set of left-censored

observations, and I is the set of interval-censored observations. Equation 1
shows how each type of censoring contributes differently to the likelihood
function.
If we can assume an appropriate distribution for the data, the use of
parametric models provides more precise estimates of the survival function
and other quantities of interest than do nonparametric estimators. Usually,
survival times are not normally distributed and models such as log-linear,
lognormal and Weibull distribution are chosen.
Klein and Moeschberger (1997) present different possible distributions
for T. To date, there are no statistical tests to compare the goodness of fit of
different parametric models used for interval-censored data. Therefore, visual
 SHELF-LIFE ESTIMATION BY SURVIVAL ANALYSIS 505

assessment of how parametric models adjust to the nonparametric estimation

is the common practice in choosing the most adequate model. Taking this into
account, the Weibull distribution was chosen. This distribution has been used
to model shelf-life studies (Gacula and Singh 1984; Cardelli and Labuza 2001)
of food; for this distribution the survival function is given by:
ln(t ) - m ˆ
S(t ) = Ssev ÊË ¯
(2)
s
where Ssev(·) is the survival function of the smallest extreme value distribution:
Ssev(w) = exp( - ew), and m and s are the model’s parameters.
The parameters of the model are obtained by maximizing the likelihood
function (Equation 1). The likelihood function is a mathematical expression
that describes the joint probability of obtaining the data actually observed on
the subjects in the study as a function of the unknown parameters of the model
being considered. To estimate m and s for the Weibull distribution, the likeli-
hood function is maximized by substituting S(t) in Equation 1 by the expres-
sion given in Equation 2.
Once the likelihood function is formed for a given model, specialized
software can be used to estimate the parameters (m and s) that maximize the
likelihood function for the given experimental data. S-PLUS (Insightful Cor-
poration, Seattle, WA, U.S.A.) survival analysis functions were used for the
Weibull parameter calculations. With these parameters the survival function
or the cumulative distribution function (F(t)) could be defined. Calculation
details can be found in Garitta et al. (2003).
To estimate the shelf life, the probability of a consumer rejecting a
product F(t) = P(T £ t) must be chosen. Gacula and Singh (1984) mentioned
a nominal shelf-life value considering 50% rejection, and Cardelli and Labuza
(2001) used this criterion in calculating the shelf life of coffee. In the present
study, shelf life was calculated for F(t) = 25 and 50%.
Sensory acceptability (measured by consumers on a 1–9 hedonic scale),
sensory data (measured by a trained panel on a 0–10 intensity scale) and
chemical data, were linearly correlated versus time. These correlations were
used to estimate those values at the shelf-life times calculated for 25 and 50%
rejection probability.
To estimate the activation energy based on the sensory off-flavor pro-
duced during storage at 20 and 35C, the following equation that combines
zero-order reaction rate (as seen in the results section this was the chosen
order) with the Arrhenius model was used:
È E Ê1 1 ˆ˘ (3)
OF = OF0 + kref ¥ t ¥ exp Í- a Á - ˜˙
Î R Ë T T ¯
ref ˚
506 A. GÁMBARO ET AL.

where OF = off-flavor at time t;

OF0 = off-flavor at time t = 0;
kref = reaction rate constant at Tref;
t = time;
Ea = activation energy;
R = gas law constant;
T = absolute temperature; and
Tref = reference temperature.

This equation is nonlinear in Ea, and to calculate its parameters (OF0, kref and
Ea), nonlinear regression facilities of Genstat 6th Edition (VSN International,
Oxford, UK) were used.

RESULTS AND DISCUSSION

Shelf-life Estimation
The maximum likelihood estimates of the parameters using the Weibull
distribution were m= 4.53, s= 0.18 at 20C, and m= 3.75, s= 0.21 at 35C.
Figure 1 shows the estimation of the cumulative distribution function
F(t) = 1 - S(t), using these parameters and Equation 2 for each temperature,
showing percentage of rejection versus time.
Table 1 shows survival analysis estimation of shelf life with their confi-
dence bands. As expected, shelf life decreases when storage temperature

Alfajores at 20 and 35C

100
35C
% of rejection

75
20C

50

25

0
0 10 20 30 40 50 60 70 80 90 100
Time (days)

FIG. 1. PERCENTAGE OF CONSUMERS REJECTING THE PRODUCT VERSUS TIME, AT 20

AND 35C
 SHELF-LIFE ESTIMATION BY SURVIVAL ANALYSIS 507

TABLE 1.
SHELF-LIFE VALUES ESTIMATED FOR A 25 AND 50% OF REJECTION BY CONSUMERS
AND THEIR RELATIONSHIP WITH ACCEPTABILITY AND OFF-FLAVOR

% of rejection Storage Shelf life (days) Acceptability Off-flavor

temperature (C) ± 95% confidence
interval Value ± SE R2 Value ± SE R2

25 20 74 ± 7 4.9 ± 0.1 0.90* 5.3 ± 0.2 0.88*

35 33 ± 4 4.7 ± 0.2 0.89* 5.8 ± 0.4 0.82*
50 20 87 ± 9 4.6 ± 0.1 0.90* 6.1 ± 0.3 0.88*
35 39 ± 3 4.2 ± 0.2 0.89* 6.8 ± 0.5 0.82*

* Significant at P £ 0.01.

increases. Shelf life for a 50% rejection would be expected to be higher than
that for a 25% rejection. However, there is a difference of 13 days at 20C and
6 days at 35C between rejection levels, and both intervals overlap when
confidence bands are considered. Weibull distribution allowed shelf-life esti-
mation with a 50% rejection of 87 days, even when the study was carried out
for 80 days.

Relationship with Acceptability, Off-flavor and Moisture Content

As both of the sensory descriptors studied (off-flavor and alfajor dryness)
behaved similarly through time, off-flavor was chosen to perform the analysis.
Off-flavor versus time and log (off-flavor) versus time were correlated to
determine reaction order, both at 20 and 35C. At 20C, R2 values were 0.88
and 0.84 for zero and first-order reaction rates, respectively. At 35C, R2 values
were 0.82 and 0.77. As the fit was slightly better for zero-order reaction rate,
this order was chosen. Off-flavor values corresponding to selected shelf-life
times can be estimated, as shown in Table 1 for 50 and 25% rejection.
Titrable acidity (expressed as percentage of lactic acid) did not change
over time (0.11–0.08% at 20C and 0.12–0.18% at 35C). Moisture content
tended to decrease as expected (12.8–11.1% at 20C and 13.0–9.7% at 35C),
but the correlation was not good and would not be of practical value in
predicting shelf life.
Acceptability measured by consumers on a 9-point hedonic scale corre-
lated highly with off-flavor and dryness variations (R2 = 0.95). This would
indicate that consumers considered these attributes in their acceptability
scores.
So as to be more conservative and assure product quality, a 25% of
rejection was selected to establish the estimated shelf life and determine
acceptability and off-flavor values. Therefore, an alfajor stored at 20C having
508 A. GÁMBARO ET AL.

an acceptability value below 4.9 and off-flavor intensity score above 5.3 would
be rejected by more than a 25% of consumers (Table 1).

Activation Energy
Using Equation 3, the activation energy ± the 95% confidence interval
was 10,850 ± 2300 cal/mol. Labuza (1982) indicated that the empirical rela-
tionship of log (shelf life) versus temperature was linear. Using this property
to calculate the activation energy from the shelf-life values estimated at 20
and 35C for a 25% rejection gave a value of Ea = 9600 cal/mol. These values
are similar to the Ea obtained from off-flavor measurements.

CONCLUSIONS

Survival analysis is a useful tool for estimating the shelf life of alfajor.
It enables the use of a relatively simple methodology, such as having 50
consumers taste alfajor samples with different storage times and answer “yes”
or “no” to whether they would consume the sample.
Acceptability and off-flavor values for a 25% of consumers rejecting the
product could be estimated from the data obtained. Chemical data (moisture
content and acidity) were not good shelf-life predictors.

ACKNOWLEDGMENTS

We are grateful for financial assistance from the Agencia Nacional de

Promoción Científica (PIP-99) and the Ciencia y Tecnología para el Desarrollo
Program (Project XI.16).

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