Journal of Food Engineering 169 (2016) 122e130

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Journal of Food Engineering

journal homepage: www.elsevier.com/locate/jfoodeng

Application of multivariate accelerated test for the shelf life estimation

of fresh-cut lettuce*
A. Derossi, L. Mastrandrea, M.L. Amodio*, M.L.V. de Chiara, G. Colelli
 di Foggia, Italy
Dip.to di Scienze Agrarie, degli Alimenti e dell'Ambiente, Universita

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 24 April 2015
Received in revised form
27 July 2015
Accepted 12 August 2015
Available online 21 August 2015

The aim of this paper was to obtain a more realistic shelf life estimation of fresh-cut lettuce by
considering several sensorial, physical and chemical attributes. The Multivariate Accelerated Shelf Life
Testing (MASLT) approach was applied, comparing the use of the traditional zero- and rst- order reactions with the Weibullian logistic model in describing score change kinetics over time. A total variance
of 79.6% was explained by two principal components (PC): PC1 (68.8%) and PC2 (10.8%). The multivariate
degradation kinetics was better described by a zero-order reaction, yielding an r2 value greater than 0.97,
while the temperature dependence of the multivariate rate constant, km, was well-tted by the loglogistic model. A cut-off criterion of
1.52 was calculated, leading to shelf life estimations of ~12.4,
~10.4 and ~3.7 days for fresh-cut lettuce stored at 0, 5 and 15  C, respectively. This method provides an
overall and accurate description of the degradation phenomena occurring during the storage of fresh-cut
lettuce.
2015 Elsevier Ltd Elsevier Ltd. All rights reserved.

Keywords:
Multivariate modeling
Principal components
Variance
MASLT approach
Shelf life

1. Introduction
Fresh-cut fruit and vegetables are minimally processed foods
with no additional treatments beyond washing, cutting and packaging. Their rapid expansion on retail markets and food service over
the last 20 years is due to the demand of ready-to-eat products with
high sensorial, nutritional and functional quality.
However, during preparation, the wounding of vegetable tissues
induce physiological disorders that signicantly decrease their
shelf-life compared to the whole product, causing an increase of
ethylene production and changes in composition, color, and rmness (Brecht, 1995; Saltveit, 1997). In addition, many variables may
modulate the physiological responses of fruit and vegetables to
wounding stress, including physiological stage, cultivar, and storage
time and temperature (Routray and Orsat, 2014; Reyes et al., 2007;
Torres-Contreras et al., 2014). Moreover, packaged product

*
This paper has not been published previously and it is not under consideration
for publication elsewhere. Moreover the text is approved by all authors and tacitly
or explicitly by the responsible authorities where the work was carried out and
that, if accepted, it will not be published elsewhere including electronically in the
same form, in English or in any other language, without the written consent of the
copyright-holder.
* Corresponding author.
E-mail address: marialuisa.amodio@unifg.it (M.L. Amodio).

http://dx.doi.org/10.1016/j.jfoodeng.2015.08.010
0260-8774/ 2015 Elsevier Ltd Elsevier Ltd. All rights reserved.

degradation reactions are affected by the evolving gas composition

inside the packages, which in turn is also affected by the temperature. Given these considerations, the prediction of the shelf-life for
this type of product has become crucial for both processors and
consumers to improve the produce logistics all over the chain and
ensure lower costs and a better nal quality of the product.
Among the methods employed to predict shelf-life, one
commonly used method is the Accelerated Shelf Life Testing (ASLT)
proposed by Labuza (1982), which is based on three main steps. The
rst one is the kinetic description of the most important degradation reactions of food at different temperatures (usually, at least
three), which may be obtained by using the common zero-, rstand second-order kinetics, as well as the cumulative form of the
Weibull model (Corradini and Peleg, 2006; Odriozola-Serrano et al.,
2009; Sothornvit and Kiatchanapaibul, 2009; Amodio et al., 2015).
Alternatively, the use of deterministic models would give important information on the pattern of reactions involved during
degradation (Tijskens et al., 2009; Lokke et al., 2012; Unuk et al.,
2012; Amodio et al., 2014), but in practice they are difcult to use
for shelf life estimation because several chemical, sensorial and
physical attributes degrade at the same time during storage,
dening the overall quality of the food product.
The second step is the denition of the temperature dependence
of the rate constants, obtained through the well-known Arrhenius
equation (Labuza, 1982) or alternatively by using the log-logistic

A. Derossi et al. / Journal of Food Engineering 169 (2016) 122e130

model (Corradini and Peleg, 2004; Derossi et al., 2010). Finally, the
parameters of these last models may be used to calculate the shelf
life of the food product. More precisely, this value commonly corresponds to the period of time to reach the limit of marketability of
the quality attribute that, among all, deteriorates faster, or it is
dened as the period of time to reach the limit of the most
important quality attribute of the product.
However, although the ASLT method enables the calculation of a
suitable shelf life estimation (Corradini and Peleg, 2004, 2007;
Derossi et al., 2010; Amodio et al., 2015), it cannot assure agreement between what is estimated and what is observed experimentally. Moreover, the denition of the cut-off criteria is rather
complicated; in addition to the use of limits dened by regulations
(i.e., safety limits), the other criteria to use are delegated by
stakeholders (i.e., scientists, processors, and consumers). In fact,
when the quality of food is dened by multiple attributes, the use of
the common ASLT may produce signicant discrepancies.
Recently, a new approach for determining shelf life by simultaneously taking into account many quality attributes was proposed by Pedro and Ferreira (2006). The Multivariate Accelerated
Shelf Life Testing (MASLT) is based on Principal Component Analysis (PCA), which nds new axes in the multivariate space to
improve the description of the experimental data structure. The
most important assumption of MASLT is that the degradation reactions are the main sources of variation in the data set, given that
the PCA is driven by the time-related phenomena. By using this
method, the PC scores may be used to build a multivariate kinetics
chart that reects the degradation information of all studied quality
attributes. Then, after a proper modeling, it is possible to estimate
the shelf life of the food product by considering the changes of all
quality attributes.
Despite the potential application of this method, it has been
used only in a few practical situations, such as for concentrated
tomato pastes (Pedro and Ferreira, 2006), for a dried apple snack
(Saavedra et al., 2013) as well as for determining shelf life through
near-infrared spectroscopy (Pedro and Ferreira, 2009). The MASLT
method has never been used for fresh-cut produce for whose
quality, more than for other food products, is rapidly affected by the
contemporaneous changes of several attributes during storage; in
addition, given that the shelf duration is usually limited to 1 or 2
weeks, an accurate estimation of its shelf life, although difcult to
obtain, is of paramount importance.
In this paper, the shelf life denition of fresh-cut lettuce was
studied by considering the degradation of a number of chemical,
physical and sensorial attributes to obtain a more accurate estimation. This enhanced accuracy will help processors improve the
produce logistics and to avoid the risk of shelf-life under- or overestimation, which will, in both cases, induce economical losses.
More specically, the work had two primary aims: (1) to apply the
multivariate accelerating shelf life testing approach (MASLT) on
fresh-cut lettuce samples and (2) to introduce the Weibullianlogistic model in the MASLT approach, comparing it with the
most traditional zero- and rst order kinetics.

123

temperature (0, 5, and 15  C): specically, 3 bags for each of the 7

sampling times (0, 1, 2, 3, 6, 8, and 11 days) plus 4 additional bags
for gas sampling. On each sampling day, the gas concentrations
were assessed before opening the bags; the physical, sensorial and
chemical quality attributes were measured as described in the
following.
2.2. Physical and sensory attributes
A group of 4 laboratory-trained panelists subjectively assessed
samples for appearance, green taste, sweetness and bitterness,
pungency, color, crispness and overall quality on a scale ranging
from 5 to 1. Appearance was scored on a 5 to 1 scale, where
5 excellent (fresh and turgid appearance, bright and uniform
color), 3 fair (slightly softening/dehydration), 1 very poor
(severe softening and presence of rot). A score from 5 to 1 was
assigned to evaluate the other attributes, such as color (5 typical;
3 beginning to brown on the cut edges; 1 severe browning on
the cut edges and vascular vessel), rmness (5 very crisp;
3 slightly softening; 1 severe softening), aroma (5 typical/
strong; 3 moderate aroma; 1 not perceived), bitterness
(5 very bitter; 3 moderately bitter; 1 not bitter) and
sweetness (5 very sweet; 3 moderately sweet; 1 not sweet).
Off-odors and off-avors were scored on a 5 to 1 scale, where
1 no off-odors/off-avors, 3 moderate off-odors/off-avors and
5 very strong off-odors/off-avors (fermentation).
The changes in the rmness of cut lettuce during storage were
determined with an Instron Universal Testing Machine (model
3340) equipped with a Kramer shear cell. Five grams of cut leaves
were placed in the Kramer shear cell and extruded with the
crosshead at a speed of 50 mm/min. Firmness was measured as the
maximum peak recorded force on the chart and expressed in
Newton (N).
Leaf lamina color was measured using a Spectral scanner (DV
s.r.l, Padova, Italy) equipped with a Spectral Imaging spectrometer
V10 type (400e1000 nm, 25 mm slit, resolution 5 nm). Color
changes were quantied in the CIE L*a*b* color space on the region
of interest (ROI). Hue angle (h arctg (b*/a*)) and saturation
(Chroma (a2b2)) were calculated from a* and b* values.
2.3. Phenolic content determination
Five grams of fresh tissue were homogenized in 2 mM NaF
methanol:water solution (80:20) for 1 min and then centrifuged at
5  C and 12,000 g/min for 10 min. The pellet was discarded and the
supernatant was retained and used as the extract. The total phenols
was determined according to the method of Singleton and Rossi
(1965). Each extract (100 mL) was mixed with 1.58 mL water,
100 mL of FolineCiocalteu reagent and 300 mL of sodium carbonate
solution (200 g L
1). After 2 h of standing, the absorbance was read
at 725 nm against a blank with a spectrophotometer (Shimadzu
UV-1700, Jiangsu, China). The total phenol content was calculated
on the basis of the calibration curves of gallic acid and expressed as
mg of gallic acid equivalents per 100 g of fresh weight.

2. Materials and methods

2.4. Modeling
2.1. Sample preparation and experimental design
Fresh Iceberg lettuce heads (cv. Aviram) were purchased from a
local grower and washed in a free chlorine solution (0.01% v/v)
before being drained and portioned into 100-g samples that were
packaged in PP bags (17.5 cm  19 cm, OTR 1800 cm3m2d
1,
WVTR 6 gm2d
1). The initial gas composition inside the packaging of 5 kPa O2 and 15 kPa CO2 followed commercially used
packaging conditions. Twenty-ve bags were stored at each

2.4.1. Theoretical background

PCA attempts to reduce the dimensionality of the experimental
data, maintaining the variability in terms of a variance-covariance
structure. PCA enables the analysis of a set of R variables, nding
a new coordinate system, C, where C < R, in which the majority of
the variability of the experimental data is captured. Moreover, the
score matrix describes the relationship between the experimental
data, while the loading matrix provides information on the

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A. Derossi et al. / Journal of Food Engineering 169 (2016) 122e130

relationships among the different dependent variables (Saavedra

and Cordova, 2011).
The ow of the mathematical steps to estimate shelf-life based
on this multivariate approach is shown in Fig. 1. The temporal
changes of the quality attributes of lettuce samples at different
temperatures were arranged in a Y m x n matrix, where m is the
number of data point collected during storage and n is the number
of quality attributes analyzed. More precisely, m is a vector (g x 1)
with g T*tT (where T is the number of storage temperature
analyzed while tT is the number of data point collected during
storage for each storage temperature), while n is a vector (1 x k)
with k the number of the quality attributes analyzed.
Then, because the quality attributes had different scales, the
experimental data were autoscaled considering the data points of
all three storage temperatures and times for each quality attribute),
obtaining a new H matrix by using the method proposed by Pedro
and Ferreira (2006).
Then, PCA was performed on the H matrix. For each of the j
principal component describing the data variability over time, a
kinetic plots including the changes of PC scores as a function of time
was obtained by separating the scores matrix (S) for each T storage
temperature. Furthermore, the loading matrix (L) was used to map

the quality attributes on PC space highlighting the relation between

them.
After identifying the number of PC that are time-related, the
changes in scores over time were modeled by the conventional
zero- and rst-order kinetics, as well as by the Weibullian model,
which improved the tting of the experimental observations of
several quality indexes as a function of time due to its extreme
exibility (Amodio et al., 2014). However, because the experimental observations in our case are the scores of time-related PC,
the following form of the Weibull model was used:

Ct C0 exp
bm Tnm T

(1)

where C0 is the PC score of fresh lettuce samples, bm(T) and nm(T)

are multivariate temperature-dependent coefcients and t (days) is
the time. As reported from Corradini and Peleg (2004), nm(T) represents the multivariate shape factor, while the reciprocal of
bm(T) is the multivariate location factor. Nevertheless, as reported
by Corradini and Peleg (2004, 2005, and 2006), because the bm
value has rate units (1/t), Eq. (1) may therefore be considered a
kinetics model. All the kinetic parameter of Eq. (1) we estimating by
using the statistical packages Statsoft ver. 10 (Tulsa, USA).
After estimating the multivariate rate constant for each kinetic
model used, their temperature dependence was studied by the
well-known Arrhenius model proposed by Labuza (1982) and by a
log-logistic model that was recently used by several authors to
estimate the shelf life of several food products (Corradini and Peleg,
2007), including fresh rocket leaves at different temperatures
(Amodio et al., 2014):

n
o
bm T loge 1 expkm T
Tcm 

(2a)

n
o
nm T loge 1 expkm T
Tcm 

(2b)

where Tcm ( C) marks the critical temperature at which the

degradation intensity increases, and Km ( C
1) is the acceleration of
the process beyond Tcm. Finally, the shelf life of samples of fresh-cut
lettuce was estimated by calculating the cut-off criteria (tc) that
represent the maximum acceptable value of the vector t (Pedro and
Ferreira, 2006):

t xa *L

(3)

tc Maxt

(4)

where xa is the raw vector containing the auto-scaled values of the

reference limits of each quality attribute that dene the threshold
of acceptability of the product, while L is the loading matrix of the
time-related PC.

Fig. 1. Flow of mathematical steps to estimate shelf-life based on the multivariate

approach.

2.4.2. Cut-off criteria for shelf life determination

To dene the cut-off criteria by Eq. (3), different reference limits
were chosen for each quality index. For sensorial attribute scores,
values of 3 and 2, respectively identied as thresholds of marketability and edibility (Amodio et al., 2015), were considered as limits
for shelf life estimation.
For hue angle, a fractional change of 8% was considered as the
limit of degradation, according to Heidmal et al. (1995), who
correlated hue angle and visual quality of Iceberg lettuce, reporting
that samples demonstrating this color variation showed severe
browning and were below the marketability limit.
Phenolic content has been widely reported to increase during
storage as a response to several types of stress (Saltveit, 2000).

A. Derossi et al. / Journal of Food Engineering 169 (2016) 122e130

Particularly for lettuce samples, the increase of phenols is related to

an increase in browning level (Campos-Vargas and Saltveit, 2002;
Murata et al., 2004). Under these considerations, the shelf-life
limit for phenol content should be dened as a maximum increase, contrarily to the other quality attributes, which normally
decrease over time. Based on a previous study of Saltveit (1997)
that reported an increase of absorbance reading for phenols of
100% (from 0.3 to 0.6 at 320 nm) signicantly reduced the visual
quality of cut lettuce by edge browning, we arbitrarily dened a
reference limit for the increase of phenols as 75% of the initial
content.
2.4.3. Statistics
An initial analysis of variance (ANOVA) was performed to evaluate the signicant differences in each quality attribute over time.
Particularly, a signicant level, p < 0.05, was used as the limit for
analyzing the differences among the samples.
The PCAs were performed by using the NIPALS algorithm. The
accuracy of the principal components was evaluated by the r2 value,
which refers to the fraction of explained variation of the experimental data.
The non-linear regressions were performed using the tting
packaged available in Statistica ver. 10 (Stat Soft, Inc., USA) through
which all computations, such as ANOVA and PCA, were performed.
3. Results and discussion
3.1. Principal components and their relationships
The analysis of variance was performed with the aim to highlight which quality attributes showed signicant variation during
storage. Sensorial scores for appearance, color, crispness, sweet
taste, bitter taste, odors, off-avor, and off-odor showed signicant
changes (p < 0.05); similarly, the rmness and hue angle among the
physical attributes and phenol content among the chemical attributes showed signicant changes (data not shown). Also gas
composition was signicantly affected by the temperature,
showing signicant differences between bags stored at high temperature, compared to the bags stored at lower temperature (0 and
5  C). Bags stored at 15  C reached at the equilibrium an atmosphere with 23% CO2 and 0.5% O2, while in the bags stored at 0 and
5  C, after an initial oxygen consumption and CO2 accumulation,
due to the stress induced by the cutting, the initial gas composition
of 5 O2 and 15% CO2 was restored.
Because samples stored at 15  C were not edible after 7 days, the
evaluation was discontinued after 9 days of storage. Samples stored
at 0 and 5  C were evaluated up to 11 days. All experimental observations were used to perform the principal component analysis.
Specically, the PCA conducted on the entire H matrix showed the
model accounted for 79.6% of the total variance of experimental
data with 2 PCs; specically, PC1 and PC2 explained 68.8% and
10.8% of the variance, respectively. The importance of the variables,
also called the variable's power, provides useful information indicating how much a variable contributes in the denition of the
principal components or how well the principal components model
that variable. Fig. 2 shows the power of the independent variables
both on the rst principal component and on the PC model. As
expected, the power of each individual variable increased with the
increasing of the number of components considered as more
variance information is included in the analysis. Firmness and offavor exhibited power values below 0.4, indicating that they
were not signicant in the denition of the PC model; in other
words, these variables did not add useful information to explain the
variance of the experimental data in the PC model. On the other
hand, the most important variable was the crispness, followed by

125

odor, sweet taste, appearance and off-odor with values always

greater than 0.90. A progressive decrease of the variable importance can be observed for bitter taste, hue angle, color and phenol
content, showing power values ranging between 0.75 and 0.89.
Moreover, the importance of the variables did not change when the
PC2 was added to the PC model, except for bitter taste which
showed a very low value when only PC1 was considered; in other
words, the PC2 mainly modeled the bitter taste, whereas the PC1
was able to model most of the dependent variables.
Fig. 3 shows the loadings of the quality attributes on the
PC1ePC2 plane. The widest difference in agreement was observed
on PC1 with the fractions of the explained variance reported above.
In addition, a signicant clustering of the variables was observed,
signifying a high correlation among several qualities attributes of
the fresh-cut lettuce. The appearance and crispness were practically overlapped, indicating an elevated correlation between the
visual aspect of the leaves and their crispness; in fact, the linear
regression of all experimental observations of these variables
showed a correlation coefcient of 0.976 and p-level < 0.01 (data
not shown). More generally, sweet taste, crispness, appearance,
color, hue angle and odor were placed close together in the
PC1ePC2 plane, indicating a good correlation among these attributes. Bitter taste also inuenced the PC model, similarly to the
above variables, but with the lowest projection on the PC1 and, on
the contrary, the highest on the positive axes of the PC2. On the left
hand side of PC1 were the phenol content, off-odors and off-avors,
indicating that they were negatively correlated with the other
quality attributes. This is obvious for off-odor and off-avor because
they indicate negative attributes that increase with time in contrast
with the positive attributes, such as color, appearance, odor, etc.,
that decrease during storage. Different remarks should be made for
the phenol content. Although phenol content may be considered a
desired attribute from a nutritional point of view, the dual role of
phenol compounds as antioxidants and as substrates for oxidative
enzymatic and non-enzymatic browning reactions is well known.
Several authors showed an increase during storage, particularly for
fruit and vegetable products (Del Caro et al., 2004; ArtesHernandez et al., 2007). In fresh-cut lettuce, in which the initial
phenol content is very low, browning is a result of an active
inductive process, requiring de novo synthesis of phenylalanine
ammonia-lyase (PAL) and the consequent accumulation of phenolic
compounds, rather than a passive oxidation of pre-existing phenols
(Saltveit, 2000). This phenomena is shown to be stimulated as a
response to several types of stress, such as wounding (Saltveit,
2000; Klaiber et al., 2005), and the increase of PAL activity is
considered the limiting factor for browning in fresh-cut lettuce
(Campos-Vargas and Saltveit, 2002; Murata et al., 2004). Accordingly with these authors, Fig. 4 shows the progressive increase of
the phenolic content of lettuce samples, which started earlier and
was more accentuated at 15  C than at the lower temperatures. For
samples stored at 0  C and 5  C, the maximum phenol content was
observed after 9 days, whereas it peaked after 7 days for the
samples stored at 15  C. The data tted the consecutive reaction
mechanism proposed by Amodio et al. (2014) well, which is based
on two main reactions, namely the de novo synthesis of phenols
promoted by PAL and the phenols degradation via polyphenol oxidase (PPO) enzymes. Correlation coefcients of 0.94 were observed
(data not shown).
On these bases, as previously reported, the shelf-life limit for
phenol content should be dened as a maximum increase rather
than the maximum reduction of the other common quality attributes. As described in the material and methods section, 75% of the
initial phenol content was considered the reference limit.
According to MacGregor and Kourti (1995) and to Saavedra et al.
(2013), the contribution plot of the PCA provides information

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A. Derossi et al. / Journal of Food Engineering 169 (2016) 122e130

Fig. 2. Power values of dependent variables on the PC model and on PC1.

1.2
1.0

bitterness score

0.8
0.6
phenols

0.4

PC2 (10.8%)

sweetness score

0.2

Firmness

off-flavor score

crispness score

0.0
-0.2

appearance
score

color score
hue angle

off-odor score

aroma score

-0.4
-0.6
-0.8
-1.0
-1.2
-1.2

-1.0

-0.8

-0.6

-0.4

-0.2

0.0

0.2

0.4

0.6

0.8

1.0

1.2

PC1 (68.8%)
Fig. 3. Loading factors describing the relationships among quality attributes of fresh-cut lettuces stored at 0, 5, and 15  C.

related to the inuence of the process variables on the scores of the

experimental observations. With the aim to analyze the impact of
each quality attribute on the overall degradation phenomena of
fresh cut-lettuce, Fig. 5 reports the contributions of the independent variables to the scores of individual samples, initially, after 11
day of storage at 0 and at 5  C, and after 9 days at 15  C (as samples
at this temperature were not evaluable after this time). Although
the contribution to the variance of lettuce samples just upon cutting was explained mainly by the aroma, a very high incidence was
also observed for sweetness, crispness, hue angle and appearance
(Fig. 5a); on the other hand, different results were observed at the
end of storage for samples stored at 0, 5 and 15  C. After 11 days at

0  C (Fig. 5b), the appearance and aroma were the most important
variables, followed by crispness. Moreover, as these attributes
decreased during storage, their contribution scores showed negative values in comparison to the fresh samples. For samples stored
at 5  C for 11 days, the color, hue angle and appearance exhibited
the greatest contribution, followed by crispness and aroma (Fig. 5c).
Finally, for samples stored at 15  C for 9 days (Fig. 5d), the crispness
and off-odors were the most important variables, followed by the
color and appearance. In addition, opposite signs were observed
according to the reduction of the crispness and the increase of offodors during aging of lettuce samples. This may indicate that a
temperature of 15  C accelerated chemical reactions and induced

A. Derossi et al. / Journal of Food Engineering 169 (2016) 122e130

127

observed on PC2. Particularly, lettuce samples stored at 15  C

showed greater variation on the PC1-axes with values ranging
between 2.82 and
6.63. Because the scores were obtained by
analyzing all quality attributes at the same time, this nding indicates that the greater overall degradation of fresh-cut lettuce
leaves occurred at 15  C and that, taking into account the result of
Fig. 5d, the major contribution to this degradation was given by the
loss of crispness and development of off-odor although other
quality attributes affected the changes of scores on PC1 axis. Fig. 6
also shows that the score changes appeared to be time-related for
both PC1 and PC2, suggesting the possibility of studying the kinetics of the overall degradation of fresh-cut lettuce samples;
however, because the higher variations were observed on the rst
components, only the PC1 scores were modeled over time with the
aim to describe the degradation kinetics of the fresh-cut lettuce
samples.
Fig. 4. Changes in phenol content of fresh-cut lettuce stored at 0, 5 and 15  C as a
function of time.

unfavorable gas condition which could have lead to fermentation

and/or microbial growth, producing undesired odor compounds
(Lopez-Galvez et al., 1997; Tudela et al., 2013).
Fig. 6 shows the score plot for fresh-cut lettuce samples in the
PC1ePC2 plane. To better illustrate the behavior of samples over
time, initial samples were labeled as F, whereas the samples at the
end of storage at 0, 5 and 15  C were respectively labeled as E
0,
E
5 and E
15. For each storage temperature, the samples exhibited
a signicant variation on the PC1, while only a slight variation was

3.2. Multivariate modeling and shelf life estimation

Table 1 reports the results of the non-linear regression of the
PC1 scores over time performed by applying the common zero- and
rst-order kinetics, as well as the Weibullian model. From the results emerged that a zero-order kinetics rate better explained the
PC1 score changes during storage, showing a correlation coefcient
always higher than 0.97; in the meantime, when the rst-order
kinetics and the Weibullian models were used, r2 values in the
ranges of 0.45e0.92 and 0.42e0.93, respectively, were obtained.
The estimated C0 values for each storage temperature were similar,
indicating that the variance of the overall quality of the fresh
samples did not affect the degradation kinetics; in fact, considering

Fig. 5. Contribution plot indicating the effects of each dependent variable on experimental observations initially (a), after 11 days at 0  C (b), after 11 days at 5  C (c), and after 9 days
at 15  C (d).

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A. Derossi et al. / Journal of Food Engineering 169 (2016) 122e130

Table 2
Estimated parameters of the Arrhenius and log-logistic models describing the
temperature dependence of the multivariate degradation rates, km, of fresh-cut
lettuce samples stored at 0, 5 and 15  C.
Arrhenius model, Ln(km) Am(Eam/RT)
Am

Condence intervals

Eam (kJ/mol)

Condence intervals

r2

25.7

12e55

6.004

0.13e25.1

0.97

Logistic model, bm Log[1 exp(Km*(TTcm))]

Fig. 6. Score plot of fresh-cut lettuce samples stored for 11 days at 0, 5  C and for 9
days at 15  C. (E
0  C), (E
5  C) and (E
15  C) indicates the samples at the end of
storage at 0, 5 and 15  C.

the condence intervals associated with each estimate, it can be

stated that no signicant differences were obtained for this
parameter. On the other hand, a progressive increase of the
multivariate rate constant, bm, was observed as the storage temperature increased with values of 0.32 d
1, 0.37 d
1 and 1.19 d
1 for
samples stored at 0, 5 and 15  C, respectively. Furthermore, from
the condence intervals, it can be observed that the differences
were signicant only when the temperature increased from 5 to
15  C. It should be considered that the degradation rates estimated
by the linear regression of the PC1 scores are the results of the
contribution of all independent variables feeding information on
the rate of the overall degradation phenomena of lettuce leaves
occurring at different storage temperatures.
Once the kinetics of the PC1 score changes over time was
described, the temperature dependence of the multivariate
degradation rates, bm, was studied by using both the traditional
Arrhenius equation and the log-logistic model. Table 2 reports the
nonlinear regression results of bm values performed by using both
the models. The Arrhenius equation tted the estimated rate constants well with a correlation coefcient of 0.97. In addition, activation energy of 6.0 kJ/mol was estimated, according to the results
of Saavedra et al. (2013) who, in studying the shelf life of dried

Km ( C1)

Condence intervals

Tcm ( C)

Condence intervals

r2

0.1403

0.18e0.46

282.4

269.0e295.7

0.99

apple snacks, reported a value for Eam of 6.544 kJ/mol. However, the
log-logistic model yielded the best equation to describe the
behavior of the multivariate rate constant, bm, as a function of
storage temperature, showing a correlation coefcient of 0.99.
Similarly, by analyzing the degradation of vitamin C in frozen
vegetables, riboavin in spinach and thiamin in red gram splits,
Corradini and Peleg (2006) showed the log-logistic model tted the
increase of rate constants as a function of temperature well.
Furthermore, Amodio et al. (2014), who studied the shelf life of
fresh rocket leaves, showed the log-logistic model was effective in
describing the temperature dependence of degradation rates. Once
the behavior of the rate constants was mathematically described,
the shelf life of fresh-cut lettuce samples may be estimated at any
temperature. Fig. 7 shows the PC1 scores as a function of time, the
ttings obtained with the zero-order model, and the cut-off criterion. According to the results of Table 1, the ts matched the PC1
scores well for all three storage temperatures, demonstrating that
the zero-order kinetics explains the multivariate changes of freshcut lettuce samples well. Then the limit values for each quality
attribute were auto-scaled as reported from Pedro and Ferreira
(2006) and then multiplied with the PC1 loading factors of the
corresponding dependent variables. To determine cut-off criteria,
the reference limits for sensorial attributes, phenolic content, and
hue angle were considered. The instrumental rmness was not
included in the calculation because a reference value for the limit of
shelf-life was not found and it was well represented by sensorial
rmness; many authors, in fact, reported a not signicant variation
over time (Foley et al., 2002; Fan and Sokorai, 2002) of instrumental
rmness or a variation of 7.5% at 9 days of storage when lettuce was
judged still marketable (Baur et al., 2004).

Table 1
Kinetic parameters estimated by non-linear regression of the PC1 scores as a function of time. Data related to fresh-cut lettuce samples stored at 0, 5, and 15  C.
Zero order kinetic, PC1 score(t) amebm*t
Temperature ( C)

am
(dimensionless)

Condence intervals

bm (d1)

Condence intervals

r2

0
5
15

2.43
2.32
2.90

1.95e2.91
1.68e2.96
1.08e4.72

0.32
0.37
1.19

0.39e0.24
0.47e0.25
1.55e0.82

0.97
0.97
0.98

First order kinetic, PC1 score(t) am*exp(bm*t)

Temperature ( C)

am
(dimensionless)

Condence intervals

bm (d1)

Condence intervals

r2

0
5
15

2.91
2.92
2.95

1.56e4.27
0.84e5.08
0.52e5.27

0.36
0.48
0.55

0.008e0.73

0.27e1.24

0.91e2.25

0.92
0.86
0.45

Weibullian model, PC1 scores(t) C0*exp(bm*tnm)

Temperature ( C)

C0
(dimensionless)

Condence intervals

bm (d1)

Condence intervals

nm

Condence intervals

r2

0
5
15

2.69
2.66
2.53

1.04e4.35
0.16e5.17
0.20e7.68

0.19
0.20
0.89

0.51/0.90

0.96/1.35

0.78/2.58

1.40
1.66
0.48

1.2/4.08

3.62/6.95

2.59/7.58

0.93
0.88
0.42

A. Derossi et al. / Journal of Food Engineering 169 (2016) 122e130

129

therefore to improve the logistics to reach the market with a better

quality product, reducing any risk of product loss due to under- or
overestimation of the shelf-life.
Acknowledgments
The authors gratefully acknowledge funding from MIUR (Italian
Ministry of University and Scientic Research) within Program PON
(Project OFRALSER, Grant Agreement PON01_01435).
References

Fig. 7. PC1 scores of fresh-cut lettuce samples stored at 0, 5 and 15  C as a function of

time. The solid red line represents the multivariate cut-off criterion.

By using the reference vector xa of the auto-scaled reference

limits previously reported, as well as the loading matrix, L, a cut-off
criterion, tc, of
1.52 was calculated through Eq. (3) and Eq. (4),
giving a shelf life of ~12.4, ~10.4 and ~3.7 days for samples stored at
0, 5 and 15  C, respectively. In comparison, when the limit of edibility (scores  2) was used as a benchmark for sensorial attributes,
a cut-off criterion of
3.0 was calculated and shelf lives of 17.0, 14.4
and 5 days were estimated for samples stored at 0, 5 and 15  C,
respectively (data not shown). The multivariate approach takes into
account all quality attributes contemporaneously because the PC1
scores were obtained from the contributions of all independent
variables on the degradation phenomena; thereby, the estimated
shelf life may be considered the results of the overall degradation of
fresh-cut lettuce leaves. This could give a more realistic shelf life
denition, particularly from a commercial point of view where the
consumers will appreciate the products on the basis of several
indexes.
4. Conclusions
For the rst time, a multivariate approach was used to obtain a
more realistic shelf life estimation of a fresh-cut product. This
approach enables the modeling of single degradation kinetics obtained by weighing the contribution of each dependent variable on
the overall quality of samples during storage. In comparison with
the most common accelerated-shelf life testing, the use of the PC
model yielded more information on the relationships among the
independent variables, as well as on their individual contribution to
the loss of quality of the lettuce leaves. This approach highlighted
that the degradation of samples stored for 9 days at 15  C was
mainly attributed to the off-odor rather than to the appearance and
color, which, on the other hand, were the most determinant for
samples stored at 0 and 5  C, respectively. Moreover, the map of
quality attributes on the PC1ePC2 plane permitted the identication of those attributes that were highly correlated, as in the case of
appearance and crispness which were practically overlapped. The
PC1 changes over time were well described by zero-order kinetics.
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described by the log-logistic model, resulting in a correlation coefcient of 0.99. Finally, the shelf life of fresh-cut lettuce was estimated by taking into account both the marketability and edibility
limits as benchmarks for calculating the cut-off criteria. This model
has potential applicability also for other cultivars and packaging
conditions, which need to be tested in order to further update the
model conditions. As results, this technique may be adapted by
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