LWT - Food Science and Technology 43 (2010) 1441e1449

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LWT - Food Science and Technology

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Freezing protocol effect on quality properties of fruit tissue according to the fruit,
the variety and the stage of maturity
Sophie Chassagne-Berces a, Fernanda Fonseca a, *, Morgane Citeau a, Michèle Marin b
a
INRA UMR 782 Génie et Microbiologie des Procédés Alimentaires, F-78850 Thiverval-Grignon, France
b
AgroParisTech UMR 782 Génie et Microbiologie des Procédés Alimentaires, F-78850 Thiverval-Grignon, France

a r t i c l e i n f o a b s t r a c t

Article history: Among the changes induced by freezing of fruits, it is still difficult to determine the major properties
Received 12 November 2008 which are the most representative of quality variation. A range of characteristics (texture, color, soluble
Received in revised form solids, water activity, water content, pH, titrable acidity) were determined before and after three
12 April 2010
different freezing protocols (at
20  C in a cold chamber, at
80  C in gas nitrogen convection chamber
Accepted 13 April 2010
and after immersion in liquid nitrogen at
196  C) for two fruit types (apple-Malus domestica Borkh- and
mango e Mangifera indica L. cv. ‘KENT’-), two varieties of apples (Golden Delicious and Granny Smith)
Keywords:
and two maturities of one variety (ripe and unripe Granny Smith). Freezing induced significant changes
Apple
Mango
mostly in texture, color and soluble solids. Property variations due to freezing depended mainly on the
Texture type of fruit, to a lesser extent on the variety of apples studied and slightly on the maturity of Granny
Color Smith apples. Among the freezing conditions tested, gas nitrogen convection at
80  C appeared to be
Physical-chemical properties the best choice for limiting fruit quality degradation.
Freezing rates Ó 2010 Elsevier Ltd. All rights reserved.

1. Introduction
Consumption of fresh fruits has remained stable for the past 20
years while that of processed fruits has recently risen considerably
(Amiot-Carlin et al., 2007). Fruit quality and specifically organo-
leptic characteristics determine consumer choice. The quality
attributes of fruits can be described as appearance, odor, texture,
taste, flavor. Still, the production of fruits is highly contingent on
area and season, and fresh fruits are perishable because of their
high water content (Torreggiani & Maestrelli, 2006). An effective
fruit preservation technique is hence required. Freezing is one of
the methods usually used for preserving food because the product
tends to maintain its original attributes, i.e., keeping its nutritional
properties as close as possible to those of fresh fruit (Canet &
Alvarez, 2006).
During freezing most of the liquid water changes into ice,
greatly reducing microbial and enzymatic activities. Oxidation and
respiration are also effectively weakened by low temperatures.
However, freezing almost always causes physical and chemical
changes in food due to this ice formation and a subsequent loss of
quality (Reid, 1996). Physical changes in frozen fruits involve
stresses in cell volume, dislocation of water, mechanical damage
* Corresponding author. Tel.: þ33 1 30 81 59 40; fax: þ33 1 30 81 55 97.
E-mail address: fonseca@grignon.inra.fr (F. Fonseca).
and freeze cracking. Many studies have focused on the effects of
freezing on textural quality of fruits including cherries (Alonso &
Canet, 1994), raspberries and blackberries (Sousa, Canet, Alvarez,
& Fernandez, 2007) and strawberries (Buggenhout, Messagie,
et al., 2006; Delgado & Rubiolo, 2005) through mechanical and/or
sensorial measurements of frozen/thawed fruits or through
microscopic measurements (Delgado & Rubiolo, 2005). Some
authors (Marin, Cano, & Fuster, 1992; Moraga, Martinez-Navarrete,
& Chiralt, 2006; Skrede, 1996) have investigated chemical modifi-
cations during freezing which are responsible for pH, soluble solid,
water content or color changes of frozen/thawed fruits. However,
only a few authors have studied physical and chemical changes
related to frozen fruit quality after freezing and only for one fruit
category at a time: apples (Bunger, Moyano, Vega, Guerrero, &
Osorio, 2004), mangoes (Botosoa, Chapleau, & Le-Bail, 2004) and
raspberries (González, de Ancos, & Cano, 2002). Moreover, since
freezing conditions were generally different and rarely character-
ized, it was difficult to compare the freezing effects.
Many works have focused on enhancing the quality of frozen/
thawed fruit through pretreatments, such as the addition of
different sugars (Chiralt et al., 2001; Marani, Agnelli, & Mascheroni,
2007) and the addition of calcium or low-methoxyl pectin
(Buggenhout, Messagie, et al., 2006; Sousa et al., 2007). Although
pretreatments helped to limit freezing degradation, they modified
the initial texture and composition of the fruit. Moreover, works on
apples revealed that untreated samples remained firmer after

0023-6438/$ e see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2010.04.004
1442 S. Chassagne-Berces et al. / LWT - Food Science and Technology 43 (2010) 1441e1449
freezing than osmodehydrofrozen samples (Tregunno & Goff, 1996).
Applying no pretreatment, Botosoa et al. (2004) studied the effect
of the freezing rate, a critical parameter for tissue damage, on
mango quality (texture and color) and showed that rapid freezing
(at
40  C) yields a better preservation of the fruit than slow
freezing (at
18  C). The study of the high pressure freezing process
focused mainly on texture improvement of frozen/thawed fruits
(Otero, Martino, Zaritzky, Solas, & Sanz, 2000) and vegetables
(Fuchigami, Kato, & Teramoto, 1997) but without considering the
other properties of fruits. Research on the impact of the freezing
process itself without pretreatment on the quality attributes of
fruits is still needed in order to better understand the complex
physical and chemical mechanisms taking place inside the fruit
tissue and improve freezing preservation.
Finally, the quality of frozen/thawed fruits depends on a large
number of factors including the type of fruit (Sousa et al., 2007), the
variety (González et al., 2002), the initial quality and the ripeness or
the time lapse between harvesting and processing (Phan &
Mimault, 1980; Plocharski, 1989). For apples, the shorter the time
from harvest to freezing, the higher the quality of frozen/thawed
fruit, whereas the quality of frozen/thawed peaches was better after
storage for 8 days (Phan & Mimault, 1980). These factors may
override the effect of the freezing process itself on quality.
The primary objective of the present work is to investigate the
effect of freezing kinetics without pretreatment on the quality
properties of fruit tissues using three freezing protocols: freezing at

20  C in a temperature controlled chamber which corresponds to
domestic refrigeration, freezing at
80  C in gas nitrogen convec-
tion and immersion in liquid nitrogen. The second objective is to
identify the most relevant quality properties (among texture, color,
soluble solids, water activity, water content, pH and titrable acidity)
that make discrimination between fresh fruits and frozen/thawed
fruits as well as the effects of the three freezing protocols possible.
We also aim at evaluating whether the relevant quality properties
rely on one or several of the following factors: the type of fruit
(apple and mango), the variety (Golden Delicious and Granny
Smith) and the maturity level (unripe and ripe Granny Smith).
2. Materials and methods
2.1. Fruits
Two fruits, apples (M. domestica Borkh) and mangoes (M. indica
L. cv. ‘KENT’), were chosen for the systematic experimental study.
Among apples, two varieties (Granny Smith-Gr and Golden Deli-
cious-Go) and two maturities for one variety (Granny Smith-Gr and
UnGr) were taken into account. The choice of the two fruits, vari-
eties and maturities was made considering their expected differ-
ences in nature, composition and quality properties. Moreover,
successful freezing of apples in terms of product quality preserva-
tion still remains a challenge to be addressed by the food industry.
The initial characteristics of the fresh apples and mangoes are
presented in Table 1. The apple varieties were purchased from an
agricultural cooperative (Dorleane, Saint Hilaire Saint Mesmin e
Loiret, France), where they were stored at 1  C under modified
atmosphere. The two apple varieties were harvested at commercial
maturity, in September 2006 for Golden Delicious and in October
2006 for Granny Smith. The Granny Smith was also characterized in
an early stage of maturity, i.e., gathered two weeks before
commercial maturity (UnGr). At laboratory, the apples were stored
in a cold chamber at 4  C, for a maximum duration of one month,
until the moment of the experiment. The mangoes (Ma) were
purchased from a local supermarket and were selected on the basis
of similar size, fruit firmness and color (yellowered). They were
then stored at ambient temperature (21  1  C), for a maximum
duration of one day before testing.
2.2. Sample preparation
The apples were placed overnight at ambient temperature
(21  1  C) before sampling. A 2 cm thick transverse cross section
was cut at the equatorial level of each apple. Cylinders (1.2 cm in
diameter and 2 cm in height) were taken equidistant from the
surface and the seed sacs in the parenchyma region using a circular
punch.
The mangoes were divided into two parts. Mango cylinders
(1.2 cm in diameter and 2 cm in height) were cut from several
transverse cross sections of 2 cm thickness equidistant from the
surface and the stone using a circular punch.
2.3. Freezing and thawing processes
Three freezing protocols were applied: at
20  C in a cold
chamber, at
80  C in a gas nitrogen chamber with high convection
(Silversas, Air Liquide, Paris, France) and by immersion in liquid
nitrogen (LN2, boiling point ¼
196  C) until the core temperature
reached the equilibrium value with the freezing temperature.
Within the freezing chamber, fruit cylinders were non adjacently
and vertically placed on a solid support. The temperature at the
cylinder core was monitored with thermocouples (type T; 0.2 mm
diameter) placed in the cylinder center. These measurements
ensured that the temperature within the samples was reached and
made it possible to estimate global freezing rates (rf) as the average
of the ratio between the temperature gap (DT) and the global
freezing time (tf) : rf ¼ Ti
Tf =tf , where Ti is the ambient
temperature (21  1  C); Tf is the freezing temperature (
20  C,

80  C or
196  C) and tf is the time to reach freezing temperature
from ambient temperature. The three protocols correspond to slow
(1  C/min), intermediate (8  C/min) and very fast (310  C/min)
freezing rates, respectively. Once frozen, the samples were packed
in polyethylene bags and thawed in a cold chamber at 4  C over-
night (usual temperature regime used in domestic refrigeration).
They were finally placed at room temperature (21  1  C) until the
sample cores reached room temperature. Fresh samples were used
as reference samples for each experimental technique. The three
freezing protocols and fresh samples are referred to as protocols in
the following sections.
2.4. Texture analysis
A TA.XT2i texture analyzer (Stable Micro Systems Ltd, God-
alming, UK), equipped with a 5 kg load cell and a force measure-
ment accuracy of 0.025%, was used to evaluate the texture of the
fresh and frozen/thawed apple cylinders. All experiments were
conducted at 21  C keeping the fruit cylinders with the same
orientation because of the fibrous non-isotropic properties of apple
flesh (Khan & Vincent, 1993). For each protocol, ten fruits and two
cylinders per fruit were analyzed resulting in 20 measurements.
Puncture tests were carried out as in Mehinagic, Royer, Symoneaux,
Bertrand, and Jourjon (2004) but with a 2 mm diameter cylindrical
probe, penetrating 10 mm into samples at 0.5 mm/s. Force vs.
distance curves were analyzed and one parameter was calculated:
firmness, defined as the flesh penetration force plateau value and
expressed in Newton. Compression tests were performed with
a 21 mm compression plate (P/21) at 0.1 mm/s until a 15% strain
was reached. Stress vs. strain curves were analyzed and Young’s
modulus (E) was obtained from the slope of the loading curve at the
point of its highest gradient.
 S. Chassagne-Berces et al. / LWT - Food Science and Technology 43 (2010) 1441e1449 1443

Table 1
Physical and physical-chemical properties of fresh Apples (Golden Delicious-Go, ripe and unripe Granny Smith-Gr) and fresh Mango cv ‘Kent’. Means across repetitions are
represented with confidence interval.

Properties Fruit Variety Maturity Measured values Literature values(a)

2
Firmness (kg/cm ) Apple Go Ripe 10  0.52 4,6-7,2 (Mehinagic et al., 2004)
6.5-6.7 (Lopez, Lavilla, Riba & Vendrell, 1998)
Gr Ripe 9.4  0.50 6-8 (Trillot, Masseron, Mathieu, Bergougnoux,
Hutin & Lespinasse, 2002)
7.3-7.8 (Lopez et al., 1998)
Unripe 14.2  0.82 /
Mango Kent Ripe 0.99  0.2 /
E (Mpa) Apple Go Ripe 2.24  0.14 1.4-2.5 (Varela et al., 2007)
Gr Ripe 2.07  0.08 2.4-4 (Varela et al., 2007)
Unripe 2.22  0.08 /
Mango Kent Ripe 0.09  0.01 /
Aw Apple Go Ripe 0.958  0.001 /
Gr Ripe 0.986  0.002 0.97 (Bunger et al., 2004)
Unripe 0.959  0.001 /
Mango Kent Ripe 0.978  0.001 /
Water content (%) Apple Go Ripe 0.858  0.004 /
Gr Ripe 0.872  0.004 0.876 (Tregunno et al., 1996)
Unripe 0.885  0.004 /
Mango Kent Ripe 0.83  0.01 0.83 (Torres, Talens, Escriche & Chiralt, 2006)
pH Apple Go Ripe 3.49  0.01 3.79 (Wu et al., 2007)
Gr Ripe 3.13  0.02 3.40 (Wu et al., 2007)
Unripe 3.08  0.02 /
Mango Kent Ripe 3.88  0.03 4.34-4.54 Other varieties (Marin et al., 1992)
Titrable acidity (g/L) Apple Go Ripe 5.36  0.32 3.8 (Wu et al., 2007)
4.85 (Mehinagic et al., 2006)
3.9-4.5 (Lopez et al., 1998)
Gr Ripe 8.04  0.27 7.3 (Wu et al., 2007)
9.3-10 (Lopez et al., 1998)
Unripe 9.38  0.26 /
Mango Kent Ripe 4.48  0.56 /
Soluble solids Apple Go Ripe 12.91  0.10 13.40 (Wu et al., 2007)
( Brix) 11.8-12.2 (Lopez et al., 1998)
12.6-15.9 (Mehinagic et al., 2006)
Gr Ripe 11.50  0.09 12.03 (Wu et al., 2007)
11.60 (Phan et al., 1980)
10.3-10.9 (Lopez et al., 1998)
Unripe 10.49  0.05 11.00 (Phan et al., 1980)
Mango Kent Ripe 16.50  0.10 15.40-17.25 Other varieties (Marin et al., 1992)
L Apple Go Ripe 80.97  0.68 /
Gr Ripe 80.06  0.52 61.64 (Bunger et al., 2004)
Unripe 79.75  0.53 /
Mango Kent Ripe 72.93  1.47 70.7-76.4 (Valente, Dornier, Piombo & Grotte, 2004)
a Apple Go Ripe -0.97  0.35 /
Gr Ripe -3.39  0.17 -2.62 (Bunger et al., 2004)
Unripe -4.43  0.24 /
Mango Kent Ripe 13.81  1.31 -3.3-1.2 Other varieties (Valente et al., 2004)
b Apple Go Ripe 26.96  1.33 /
Gr Ripe 16.55  0.78 18.30 (Bunger et al., 2004)
Unripe 18.76  0.87 /
Mango Kent Ripe 57.59  3.78 43.8-51.1 (Valente et al., 2004)
(a)
values recalculated from the literature data

2.5. Measuring amounts of soluble solids, pH and titrable acidity 2.6. Dry matter content

For each protocol, the amount of soluble solids, pH and titrable
acidity were determined using fruit juice squeezed from 10 fruit
cylinders.
The soluble solids were measured with a digital refractometer
(Atago type ABBE, Itabashi-ku, Tokyo, Japan). The results were
reported as in Brix degrees ( Brix  0.1%) at 21  C  1  C. Titrable
acidity was measured by titrating the fruit juice with 0.1 mol/L
NaOH up to pH 8.1 using a digital pH meter (TitroLine easy, Schott
Instrument, Germany) with measurement accuracy of 0.1%. The pH
was determined at 21  C  1  C before titration. The results were
expressed in gram-equivalents of malic acid or citric acid per liter of
juice, respectively, for apples or for mangoes. Measurements were
performed eight times.
Determinations were made on 5 g of fruit (two cylinders) by
drying samples in an oven, at 80  1  C for 48 h under atmospheric
pressure, fitted on AOAC method 20.013. Samples were weighed
with a precision scale of 0.001 g. For each protocol, ten fruits and
four cylinders per fruit were analyzed.
2.7. Color
The color of samples was measured using a spectrophotometer
(Minolta Co., Ltd France) at 20  C. Results were expressed in the
L*a*b* scale with an accuracy of 0.1%, where L* is the lightness, a*
represents the greenered color axis (redness) and b* the blueeyellow
axis (yellowness), as recommended by the International Commission
1444 S. Chassagne-Berces et al. / LWT - Food Science and Technology 43 (2010) 1441e1449
on Illumination in 1976. In order to include all color parameters in one
parameter, the total color differences (DE) ¼ (DL2 þ Da2 þ Db2)0.5 were
also calculated with the fresh state as a reference. For each protocol,
ten fruits and two cylinders per fruit were analyzed.
2.8. Water activity
Aw was measured at 21  1  C with an Aw-meter FA-st/1 (Food
Analysis Science and Technology, GBX Scientific Instruments,
Romans sur Isère, France) presenting an accuracy of 0.003 Aw. For
each protocol, cylinders from five fruits with one replication were
analyzed.
2.9. Imaging ice crystals into cells
Two apples and two cylinders per fruit were studied for each
freezing protocol. Frozen samples at
20  C and at
80  C were
fixed by immersion in liquid nitrogen. This step was not necessary
for the last freezing protocol corresponding to immersion in liquid
nitrogen. No changes in ice structure due to fixation were apparent.
Frozen apple cylinders were transferred into the cold stage of
a cryosystem (GATAN) for scanning electron microscopy (PHILIPS
525) and then cut with a knife (at
150  C). After etching at
80  C
to reveal cell outlines, cylinders were cooled at
160  C before
being coated with gold, and the surface was examined at
160  C
under a low acceleration voltage (10 keV).
2.10. Data analysis
Based on a large quantity of experimental data on fresh and
frozen/thawed fruits (5e10 fruits and 8e20 measurements for each
quality property), analysis of variance (ANOVA) was applied
according to the general linear models procedure of the SAS
program, version 9.1 (SAS Institute, Inc., Cary, NC, USA). The level of
significance was set for P < 0.01.
In order to facilitate data analyses, the percentage of variation
(x) between the average quality property values in the frozen/
thawed state (Val.frozen) and the average quality property values in
the fresh state (Val.fresh) was calculated according to the following
formula:
 
Val:frozen
Val:fresh
xð%Þ ¼  100:
Val:fresh
In an initial step, an ANOVA was carried out to identify the
quality properties that make it possible to discriminate between
fresh and frozen/thawed products, for different groups of data,
taking three factors into account: the fruit, the variety and the
maturity. The ANOVA assessed the main effects (before and after
freezing, fruit, variety and maturity) and some interactions (free-
zing*fruit, freezing*variety, freezing*maturity) on quality proper-
ties. This analysis was performed for each of the three freezing
protocols studied. In a second step, the same approach was used to
identify the quality properties making differentiation of the three
freezing rates considered possible. In all analyses, when significant
quality property differences were observed (P < 0.01), quality
property intensities were compared using the Student Newman
Keuls (SNK) multiple comparison test.
3. Results & discussion
3.1. Characterization of fresh state
In general, the characteristics measured on fresh mangoes and
fresh apples varied in the same range of values as the literature data
(Table 1), thus validating the different analytical methods applied
to the fruit cylinders used in this work. As expected, all quality
parameters made it possible to distinctively characterize apples
and mangoes. Fresh apples exhibited higher values of firmness,
water content, titrable acidity and lightness but lower values of
soluble solids ( Brix), redness and yellowness than fresh mangoes.
However, some properties were not relevant for discriminating
apple variety (texture and lightness) or maturity level (pH and
lightness). In previous works (Mehinagic et al., 2004; Varela,
Salvador, & Fiszman, 2007), Granny Smith was observed firmer
than Golden Delicious, but this discrepancy may be explained by
differences in cultivation practices and climatic conditions. The
soluble solids and the pH were higher for Golden Delicious than for
Granny Smith, in agreement with literature (Wu et al., 2007). A loss
of texture (Firmness and Young’s modulus) was observed for ripe
Granny Smith apples compared to unripe Granny Smith apples.
This softening is a consequence of maturation and has already been
instrumentally measured on York Imperial and Golden Delicious
apples (Ingle, D’Souza, & Townsend, 2000; Mehinagic, Royer,
Symoneaux, Jourjon, & Prost, 2006). Increasing values of soluble
solids ( Brix) and pH during maturation have been observed for
Granny Smith, in agreement with results on Golden Delicious
(Mehinagic et al., 2006).
These quality criteria according to the type of fruit, the variety
and the maturity of fresh samples may contribute to a better
understanding of quality variations observed after freezing.
3.2. Freezing effect on texture parameters
Whatever the product studied and the freezing protocol applied,
the values of the two texture parameters studied decreased
significantly (P < 0.01) after freezing (Fig. 1). It is well established in
the literature that freezing affects texture of both mangoes (Botosoa
et al., 2004; Chiralt et al., 2001) and apples (Marani et al., 2007;
Phan & Mimault, 1980). This work pointed out that the impact of
the freezing protocols on the two texture parameters analyzed was
much higher for apples (mean variations of 86% on firmness, and
98% on Young’s modulus) than for mangoes (mean variation of 62%
on firmness and between 85% on Young’s modulus). These results
can be related to the difference of texture between apples and
mangoes in the fresh state: the higher the firmness of the fresh
fruit, the higher the texture degradation observed after freezing. In
order to better understand the textural behavior of apples and
mangoes, typical examples of forceedistance curves obtained by
puncture test for frozen/thawed samples are shown and compared
with those obtained for fresh apples (Fig. 2) and mangoes (Fig. 3a),
respectively. For apples, freezing provoked a modification of
behavior shown by the apparition of a maximal force on the curves
of frozen/thawed samples (arrows), but the force diminished and
reached a lower constant value (Fig. 2). This textural profile for
apple can be explained by two tissue behaviors, suggesting freezing
gradient between outer layer and cylinder core: a quite rigid
external layer which had to be crossed by the punch before large,
homogeneous and less rigid part could be reached. This outer layer
conferred a higher deformability to frozen/thawed samples,
resulting in a longer distance at the rupture force compared to the
fresh tissue. For mangoes, less sensitive to freezing, the measured
force tended to remain constant following initial tissue failure both
in the fresh state (F) and in the frozen/thawed state (indicated by 8,
2, LN2 in Fig. 3a). However, measured force values were very low
and the limits of texture analyzer sensitivity were certainly reached
especially for the short distances. Observing the results from the
compression test (Fig. 3b) made it possible to differentiate the
textural behaviors of mango after freezing/thawing, highlighting
the interest of coupling compression test with puncture test in the
 S. Chassagne-Berces et al. / LWT - Food Science and Technology 43 (2010) 1441e1449
Fig. 1. Effect of freezing protocols at
20  C ( ), at
80  C ( ) and immersion in liquid
nitrogen ( ) on two texture parameters (a-Firmness and b-Young’s Modulus) for
Mango (Ma), Golden Delicious (Go), ripe Granny Smith (Gr) and unripe Granny Smith
(UnGr). Fresh samples ( ) were used as control. Means across 20 repetitions are
represented with confidence interval.
case of lower force values to visualize behavior through short
distances.
Concerning apple varieties (Fig. 1), Granny Smith exhibited
lower firmness degradation (54%) than Golden Delicious (80%) in
the frozenethawed state in particular when frozen at
80  C. A
higher maturity reduced the degradation observed on firmness
following freezing. The ripe Granny Smith (Gr) exhibited firmness
degradation about two times lower than that of the unripe Granny
Smith (UnGr). As a conclusion, the texture parameter variation due
to freezing was strongly linked to the type of fruit, to a lesser extent
to the variety of apples and slightly to the maturity level of Granny
Smith. These results are in agreement with other works reporting
texture degradation for different fruits. Accordingly, raspberry
exhibited about three times higher texture degradation than
blackberry (Sousa et al., 2007). On the contrary, variations in
texture after freezing between four varieties of raspberries
(González et al., 2002) differed only about 1.5 times. Finally, texture
degradation was slightly different between two maturity levels of
Granny Smith apples or peaches (Phan & Mimault, 1980).
The two texture parameters also appeared to be relevant
properties to discriminate the three freezing protocols studied. For
apples, puncture (firmness) and compression (Young’s modulus)
tests revealed freezing at
80  C as the protocol making the lowest
texture degradation possible. For the two other freezing protocols,
a discrepancy appeared according to the texture measurement
applied: freezing at
20  C caused a higher firmness and a lower
1445
Fig. 2. Forceedistance curves obtained during puncture test for fresh samples (F) and
thawed samples after freezing: at
20  C (2), at
80  C (8) and immersion in liquid
nitrogen (LN2). (a) Golden Delicious, (b) ripe Granny Smith and (c) unripe Granny
Smith. Arrows indicate the maximal force.
Young’s modulus than freezing by immersion in liquid nitrogen.
This result can be explained if we consider that each texture
analysis method assesses different textural properties of the fruit
tissue. Puncture test evaluates local fracture behavior (Bourne,
2002, p. 427). Compression test is a general measurement of the
deformability of the tissue taken as a whole (Bourne, 2002, p. 427).
Freeze cracking following immersion in liquid nitrogen is well
known to produce an undesirable fracturing of fruits and vegeta-
bles, and it occurs particularly when the sample has a relatively
large volume, water content and porosity (Kim & Hung, 1994). Such
is the case for the experimental apple cylinders used in this work
and the final consequence is the highest loss of firmness after
immersion in liquid nitrogen assessed mainly by puncture test.
Images of carrot tissue frozen by slow freezing conditions showed
more elongated and more irregular “tissue particles,” including
cells and intercellular spaces, than fresh tissue. In turn, images
1446 S. Chassagne-Berces et al. / LWT - Food Science and Technology 43 (2010) 1441e1449
Fig. 3. Forceedistance curves (a) obtained during puncture test and Stressestrain
curves (b) obtained during compression test for fresh mangoes (F) and thawed
mangoes after freezing: at
20  C (2), at
80  C (8) and immersion in liquid nitrogen
(LN2).
obtained by cryogenic freezing presented “tissue particle” shapes
similar to fresh tissue particles (Buggenhout, Lille, et al., 2006).
These structure modifications are certainly better reflected in the
compression test, resulting in a higher reduction of Young’s
modulus after freezing at
20  C than after immersion in liquid
nitrogen. Moreover, it is accepted that a fast freezing rate, corre-
sponding to immersion in liquid nitrogen, better preserves local
structure. Fast freezing induced the formation of a large number of
small ice crystals (Fig. 4a) and left an important fraction of unfrozen
water, while slow freezing, corresponding to freezing at
20  C, led
to fewer ice crystals but of larger size (Fig. 4b). The low dehydration
associated with small ice crystals induced less breakage of cell
walls, and consequently less texture deterioration than the slow
freezing rate (Mazur, 1984). Moreover, the thawing rate compared
to the freezing rate generally emphasizes the degradation which
has already taken place in the frozen state. The differences
observed between the two fastest freezing rates (8  C/min at

80  C and 310  C/min for immersion in liquid nitrogen) could be
also explained by greater differences between freezing and thawing
rates for immersion in liquid nitrogen (100 times) than at
80  C (8
times). Ice crystals which are formed rapidly during immersion in
liquid nitrogen can then grow faster during thawing than after
freezing at
80  C. The migratory recrystallisation of ice, which
takes place when molecular mobility increases, makes the crys-
tallization of residual water that is kinetically inhibited during
cooling possible.
For mangoes, only the compression test (Young’s modulus)
made it possible to discriminate three freezing protocols (Figs. 1
and 3). Freezing at
80  C led again the lowest texture degrada-
tion while no significant differences were observed following
freezing at
20  C and by immersion in liquid nitrogen.
Fig. 4. Cryo-Scanning Electron Microscopy images of Golden Delicious tissue frozen in
two different protocols. (a) Freezing by immersion in liquid nitrogen; (b) Freezing at

20  C. Ice crystals are arrowed. Scale bar ¼ 200 mm.
3.3. Freezing effect on color parameters
Changes of color, representative of important browning or
darkening of fruit tissues, were significant after freezing and
thawing (Fig. 5), as a consequence of no pretreatment. In the case of
apples, lightness decreased (Fig. 5a), the redness and the yellow-
ness increased after freezing (Fig. 5b and c, respectively), in
agreement with previously reported study in Golden Delicious
(Mastrocola, Pittia, & Lerici, 1996). In the case of mangoes, redness
decreased after freezing (Fig. 5b). Similar results were found in
previous work on other variety of mangoes (Botosoa et al., 2004).
The impact of freezing on coloration parameters was higher for
apples (mean variations of about 28% on lightness) than for
mangoes (mean variations of about 3% on lightness). The chromatic
difference DE (Fig. 5d) confirmed these results varying from 11 for
mangoes to about 30 for apples. The oxidized chemical compounds
formed following freezing differed from mangoes to apples. Thus,
the color changes could be attributed to carotenoïds (fat soluble) in
the case of mangoes and to polyphenols (water soluble) for apples
(Skrede, 1996). Polyphenol compounds are known to be more
sensitive to enzymatic browning reactions than carotenoids, thus
explaining the more pronounced color changes (to brown)
observed for apples than for mangoes. Changes of color parameters
after freezing and thawing depended as well on the variety (Golden
Delicious-Go and Granny Smith-Gr). Variations of DE values were
 S. Chassagne-Berces et al. / LWT - Food Science and Technology 43 (2010) 1441e1449
Fig. 5. Effect of freezing protocols at
20  C ( ), at
80  C ( ) and immersion in
liquid nitrogen ( ) on color parameters (a-Lightness, b-Redness, c-Yellowness and
d-DE) for Mango (Ma), Golden Delicious (Go), ripe Granny Smith (Gr) and unripe
Granny Smith (UnGr). Fresh samples ( ) were used as control. Means across 20
repetitions are represented with confidence interval.
1447
36% higher for Granny Smith than for Golden Delicious (Fig. 5d).
The impact of freezing on color parameters was the same for ripe
(Gr) and unripe Granny Smith (UnGr).
Among color parameters, the chromatic differences (DE) and
lightness variations on apples differed slightly according to the
three freezing protocols. Immersion in liquid nitrogen induced the
lowest lightness changes (mean variation of 24%) among apple
varieties (Go and Gr in Fig. 5a). The chromatic difference (DE)
variation (Fig. 5d) confirmed these results varying on average from
27 after liquid nitrogen immersion to 31 after freezing at
20  C or
at
80  C. No significant differences in color parameters between
freezing protocols were observed for the unripe Granny Smith
(UnGr). For mangoes, redness exhibited differences according to
the freezing protocol studied. However, taking into account the
high confidence intervals on redness and the similar effect on
lightness, yellowness and chromatic differences, color parameters
did not make it possible to clearly discriminate the freezing
protocols for mangoes.
3.4. Freezing effect on physical-chemical parameters
For apples, the water content decreased significantly (P < 0.01)
after freezing/thawing (mean variation of 2%) (Fig. 6a). For
mangoes, no significant changes in the water content occurred after
freezing/thawing, in agreement with results of Marin et al. (1992)
on other mango varieties. No significant variations of water
content between apple varieties, maturity levels and freezing
protocols were observed. Water content appeared thus to be rele-
vant to discriminate the frozen/thawed states from the fresh states,
nevertheless it did not make it possible to discriminate the different
freezing protocols.
The water activity did not significantly change after the different
freezing protocols applied to apples and mangoes compared to the
fresh states.
The soluble solids ( Brix) of apples increased significantly
(P < 0.01) after freezing whereas those of mangoes did not change
significantly whatever the freezing protocol (Fig. 6b). For mangoes,
similar results were found in other mango varieties (Marin et al.,
1992). Considering only apple varieties, variations of soluble
solids after freezing/thawing were 2e3 times higher for Granny
Smith than for Golden Delicious. A higher maturity can slightly
increase the effect of freezing on  Brix variations, ranging from 2%
to 12% for ripe Granny Smith vs. from 3% to 8% for unripe Granny
Smith.
The soluble solids also appeared to be relevant to discriminate
the different freezing protocols. On apples, liquid nitrogen
immersion showed lower variation of soluble solids (lower than
2%) than freezing at
20  C or at
80  C. Since water content did
not discriminate freezing protocols, the variations of  Brix
following freezing cannot be ascribed to a concentration of soluble
solids. In turn, the cell wall degradation due to ice crystal formation
can induce soluble solid extraction (especially sugars) from cells.
Indeed, slow freezing rates are well known to produce fewer ice
crystals but of larger size which may degrade the cell structure of
the product (Fennema, 1975) and thus facilitate the extraction of
sugars.
After freezing pH value changes differed among the types of
fruit (Fig. 6c) while the values of total acidity did not significantly
change after freezing. For apples, pH increased after freezing/
thawing. For mangoes, pH values decreased after freezing/thawing.
For both fruits, even if the pH changes were significant (P < 0.05),
they remained minor (around 2%), in agreement with previous
works on mangoes (Marin et al., 1992) and raspberry varieties
(González et al., 2002). From these results, whatever the fruit and
1448 S. Chassagne-Berces et al. / LWT - Food Science and Technology 43 (2010) 1441e1449
Fig. 6. Effect of freezing protocols at
20  C ( ), at
80  C ( ) and immersion in
liquid nitrogen ( ) for Mango (Ma), Golden Delicious (Go), ripe Granny Smith (Gr) and
unripe Granny Smith (UnGr) on physical-chemical properties: (a) water content, (b)
soluble solids ( Brix) and (c) pH. Means across repetitions are represented with
confidence interval. The number of repetitions varied according to the property: 20
repetitions for water content, 8 repetitions for soluble solids and 3 repetitions for pH.
Fresh samples ( ) were used as control.
the freezing protocols, pH and total acidity did not make it possible
to discriminate the fresh state from the frozen/thawed state.
4. Conclusion
From the results concerning the freezing effect on different
quality parameters, texture (firmness and Young’s modulus), color
(DE and redness), soluble solids and water content appeared as the
most relevant properties to discriminate the frozen/thawed states
from the fresh states, whatever the fruit, the variety and the
maturity studied. For differentiating freezing protocol effect,
texture and soluble solids proved to be the most appropriate
quality parameters.
Because of the different structural modifications occurring
inside the fruit tissues, the two texture measurements (puncture
and compression tests) appeared to be complementary and should
be used to better understand the freezing effect.
Mango proved to be a more suitable fruit for freezing than apple.
The lower modification during freezing of mango texture, color,
soluble solids and water contents can be related to the lower initial
rigidity of mango compared to apple. This work confirmed that
structure preservation is essential to maintaining the other quality
parameters of fruits after freezing.
Freezing at
80  C could be considered the best choice among
the conditions tested for freezing small fruit cylinders since the best
firmness preservation (lowest firmness loss) is currently associated
to higher sensorial quality, thus conditioning acceptability by the
consumer.
Based on this systematic global approach which highlighted
differences in fruit tissue texture, color and soluble solids, it would
be interesting to go forward with the study of the structure at cell
scale (cell wall composition, microscopic observation) in order to
better understand the stress mechanisms involved during freezing.
Moreover, by using this approach to study different freezing
kinetics, coupled to sensorial acceptability tests, it should be
possible to define optimal thermal itinerary for freezing fruits.
Acknowledgements
The authors wish to thank Air Liquide which financially sup-
ported this research and Dorleane SA who furnished apples and
made their storage under a modified atmosphere possible. We
would also like to thank Catherine Renard and Barbare Gouble
(INRA, SQPOV, Avignon) and Maud Panouillé (INRA, UMR 782
GMPA, Thiverval-Grignon) for fruitful discussion. Thank you to
Donald White for revising the English version of the manuscript.
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