Accepted Manuscript

Title: Kinetic study of the effect of the osmotic dehydration

pre-treatment with alternative osmotic solutes to the shelf life
of frozen strawberry

Author: Efimia K. Dermesonlouoglou Maria Giannakourou

Petros S. Taoukis

PII: S0960-3085(16)30038-4
DOI: http://dx.doi.org/doi:10.1016/j.fbp.2016.05.006
Reference: FBP 718

To appear in: Food and Bioproducts Processing

Received date: 17-4-2015

Revised date: 22-4-2016
Accepted date: 17-5-2016

Please cite this article as: Dermesonlouoglou, E.K., Giannakourou, M., Taoukis,
P.S.,Kinetic study of the effect of the osmotic dehydration pre-treatment with alternative
osmotic solutes to the shelf life of frozen strawberry, Food and Bioproducts Processing
(2016), http://dx.doi.org/10.1016/j.fbp.2016.05.006

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 HIGHLIGHTS

Osmotic dehydration as a prefreezing treatment improved frozen strawberry quality.

Alternative osmotic solutes (carbohydrates) were used.

Frozen strawberry quality (vitamin, color, texture, driploss, sensory) was studied.

Vitamin C loss and colour change of frozen strawberry were mathematically modelled.

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Developed models can be used for the prediction of quality and shelf life.

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Page 1 of 43
 Kinetic study of the effect of the osmotic dehydration pre-treatment with alternative

osmotic solutes to the shelf life of frozen strawberry

Efimia K. Dermesonlouogloua*, Maria Giannakouroub, Petros S. Taoukisa

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a
National Technical University of Athens, School of Chemical Engineering, Laboratory of

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Food Chemistry and Technology, 5, Iroon Polytechniou, Zografou, 15780,

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Athens-Greece

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b
Technological Educational Institute of Athens, Faculty of Food Technology and Nutrition,

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Department of Food Technology, Agiou Spyridonos Str., Egaleo, 12210, Athens-Greece
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*E-mail Address: efider@chemeng.ntua.gr; Tel No: +30 210 772 3118; Fax No: +30 210
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772 3166
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Page 2 of 43
 Abstract

Quality of frozen strawberry can significantly be improved by means of osmotic

dehydration as a pre-freezing treatment. The objective of this work was to study the effect of

osmotic pre-treatment with alternative osmotic solutes, oligofructose and a high DE

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maltodextrin, on the quality and sensory properties of frozen strawberry tissue. Vitamin C,

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colour, texture, drip loss after thawing and sensory characteristics of pre-treated and

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conventionally frozen samples were kinetically studied at isothermal conditions, and their

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temperature dependence was modelled by the Arrhenius equation. Dehydrofrozen samples

exhibited a significantly improved vitamin C stability especially at temperatures, below –12

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°C. The cryostabilization result of the oligofructose could be related to the glass transition

behaviour of the modified osmo-dehydrofrozen food matrix. Colour deterioration was more
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intense in the case of untreated frozen strawberry samples. Osmo-dehydrofrozen compared to

conventionally frozen strawberry halves showed an improved firmness for prolonged storage
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period and less water loss during thawing maintaining the strawberry tissue integrity. Sensory

evaluation confirmed a good organoleptic quality in osmo-dehydrofrozen strawberries. The

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shelf life of all samples was calculated based on vitamin C (30 % loss) and colour (50 %
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chroma index retention) kinetic study results.

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Keywords

Fragaria x ananassa, osmotic dehydration, alternative carbohydrates, frozen storage,

vitamin C, colour

Page 3 of 43
1. Introduction

Strawberries (Fragaria x ananassa) are rich in nutrients, such as amino acids and

vitamins (Souci et al., 2000; Cheng et al., 2014), and highly valued due to their sensory

properties, particularly sweetness and aroma attributes but are highly perishable owing to

their high physiological post-harvest activity, making their effective management a challenge

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(Garcia et al., 1998; Han et al., 2004; Blanda et al., 2009; Bodelón et al., 2013; de Brujin and

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Borquez, 2014). Among the berry species, strawberries have been in the center of commercial

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interest due to their superior sensory characteristics, in fresh or processed forms, such as

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juices, liqueurs, sorbets, ice cream, concentrates, and dairy products (Moreno et al., 2000;

Garcia-Palazon et al.,2004). In order to maintain strawberry quality and prolong its shelf life
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and season availability, freezing is one of the common widely applied preservation methods

(Bouzari et al., 2015), causing however, several undesirable changes to this sensitive tissue
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(Blanda et al., 2009; Oliveira et al., 2015), such as texture degradation, colour alteration and

nutritional loss (Martinez-Navarrete et al., 2001; Sulaiman and Silva, 2013; Oliveira et al.,
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2015). According to Mohammad et al. (2004), who studied the effect of freezing rates on

quality reduction of frozen strawberries, most of the changes during storage period are related
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to vitamin C loss and colour change. Polyphenoloxidase (PPO) present in strawberry tissues,
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causes a red colour loss because of deterioration of anthocyanin pigments and browning

(Holcroft and Kader, 1999; Watanabe et al., 2011). Changes in anthocyanins seem to be one
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of the causes of fruit browning, and their degradation is the result of coupled oxidation by

PPO in the presence of other phenolic compounds (Zhang et al., 2000; Nunes et al., 2005;

Holzwarth et al., 2013).

Osmotic dehydration prior to freezing (osmodehydrofreezing) has been used to

produce several kinds of fruits and vegetables that can be stored for long periods of time with

a good retention of texture and colour (Sormani et al., 1999; Dalla Rosa and Spiess, 2000;

Page 4 of 43
Maestrelli et al., 2001; Dermesonlouoglou et al., 2007a, b, 2008; Blanda et al., 2008a, b,

2009; Cheng et al., 2014). During osmotic processing, water flows from the product into the

osmotic solution, while osmotic solute is transferred from the solution into the product with a

determined ratio depending on process conditions (Lucas and Raoult-Wack, 1998; Fito et al.,

2001; Kowalska and Lenart, 2001; Peiro-Mena et al., 2006, 2007). Osmotic pre-concentration

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has been proposed as an effective pre-treatment in combined preservation techniques as well

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as a pre-treatment for producing fresh-like, minimally processed products, selectively

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enriched (Raoult-Wack, 1994; Torreggiani and Bertolo, 2000; Castello et al., 2006, 2010;

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Nuñez-Mancilla et al., 2011, 2013; Radziejewska-Kubzdela et al., 2014).

During osmotic treatment of strawberries, changes may occur at any level and involve
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the cell structure of fruits, their chemical composition (Chiralt and Talens, 2005; Rizzolo et

al., 2007), changes of main sensory attributes and volatile compounds (Chiralt et al., 2001;
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Talens et al., 2002). The use of dehydrofreezing for obtaining frozen fruit with a high

nutritional value and favourable sensory qualities has been reported by Dermesonlouoglou et
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al. (2007a, b,2008) and Blanda et al. (2008a, b; 2009). Although quality loss of frozen

strawberries has been previously reported (Dermesonlouoglou et al., 2004; Blanda et al.,
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2009), a thorough and systematic kinetic study of the most important quality indices, at a
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wide temperature range, is not currently available. In this study, strawberries, untreated and

pre-treated with the proposed, alternative osmotic solutions (using oligofructose and a high
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DE maltodextrin) were kinetically studied in the temperature range of practical interest.

The aim was to use low molecular weight carbohydrates of special functional and/or

nutritional characteristics that do not alter significantly the sensory characteristics of the

osmo-preconcentrated tissue. Oligofructose is a non-digestible oligosaccharide, with

exceptional dietary fiber properties and prebiotic activity (De Gennaro et al., 2000; Rao,

2001). It is officially recognized as a natural food ingredient and is classified as a dietary

Page 5 of 43
fiber in almost all European countries (Roberfroid, 2000). Furthermore, an additional target

was to study a variety of quality attributes, responsible for strawberry degradation and decide

the most appropriate one for defining its end of shelf life. Finally, the effect of the alternative

agents proposed instead of the traditional carbohydrates was investigated to assess the quality

retention of the osmosed tissue.

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2. Materials and Methods

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2.1. Sample preparation

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Fresh strawberries, obtained directly from a producer from Southern Greece were

transported to the laboratory 24h within harvesting. Strawberries were sorted, washed and cut
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into halves (of approximately 8 g weight). The average moisture content (WC) the fresh

strawberry halves was 93.8 % on a dry weight basis; the average water activity value (aw) was
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0.990 (Aqua LAB 4TEV, Decagon Devices, Inc., USA); pH and total soluble solids (TSS)

content of fresh strawberries were 4.0 and 5.1 Brix (average values), respectively.
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2.2. Solution preparation

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Solutions of 56.5 % carbohydrate concentration were prepared by blending with

distilled water on a weight-to-weight basis. The carbohydrates used were, glucose

(C*Dex2001, Cerestar, Italy) and two alternatives, oligofructose (RAFTILOSE, Orafti,

Belgium) and a high DE (Dextrose Equivalent= 47) maltodextrin coded as HDM

(GLUCIDEX47, Roquette, France). The selection of specific carbohydrates in high

concentrations was based on a previous comparative study of alternative osmotic agents

Page 6 of 43
(Dermesonlouoglou et al., 2007a,b, 2008). Selected osmotic solutes led to the optimum mass

exchange (water loss and solid gain) and sensory characteristics of the final product. NaCl

(3.5 % wt/wt) and CaCl2 (1.5 % wt/wt) were also added for the improvement of the sensory

characteristics. Small additions of sodium chloride NaCl to the osmotic solution has been

shown to increase the driving force of the process and also attenuate the sweetness of fruit

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(Lerici et al., 1985). Calcium chloride CaCl2 is used to minimize tissue damage during

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processing by interaction with pectins and other cellular wall components which reinforces

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mechanical properties of plant cellular matrix (Gras et al., 2003).

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2.3. Osmotic pre-treatment
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Strawberry halves were subjected to osmotic dehydration at a mild temperature35°C.
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Samples were removed from the osmotic solution at 150 min time, blotted gently by a tissue

paper in order to remove adhering water and then weighed. The ratio of strawberry halves to
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osmotic solution was 1:5 (wt/wt). The time-temperature combination of 150 min-35 °C

selected for the following shelf-life kinetic study led to the most desirable water loss/solid
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uptake values and the most acceptable sensory characteristics.

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The osmotic pre-treatment lowered water activity of strawberry samples, down to a

0.97 level (aw of osmotically pre-treated strawberry samples with the osmotic solutes glucose,
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0.9710.006, oligofructose, 0.9770.005, and HDM, 0.9800.006). Sample total soluble

solids TSS content increased after pre-treatment with a corresponding decrease in the

moisture content, WC (TSS of non-treated strawberry samples, 5.10.4 º Brix; osmotically

pre-treated strawberry samples with the osmotic solutes glucose, 15.13.8 º Brix,

oligofructose, 12.81.5 º Brix, and HDM, 13.02.0 º Brix, and WC of non-treated strawberry

Page 7 of 43
samples, 0.9380.028; osmotically pre-treated strawberry samples with the osmotic solutes

glucose, 0.8330.052, oligofructose, 0.8520.024, and HDM, 0.8540.017 g water/g).

2.4. Freezing

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Strawberry halves osmodehydrated at a temperature of 35 °C for 150 min as well as

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untreated samples, were quick frozen at -40°C by a forced air convection [h=11 W/(m2*K) -

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Sanyo MIR 553, Sanyo Electric Co, Ora-Gun, Gunma, Japan], packed in pouches out of

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BOPP-PE laminate film used for commercial frozen vegetable products, and kept at this

temperature for a short period of time before being distributed into controlled temperature

cabinets.
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2.5. Shelf life kinetic study
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Packages of untreated, and pre-treated containing the osmotic solute frozen strawberry

samples were stored in controlled temperature cabinets (Sanyo MIR 153, 253 and 553, Sanyo
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Electric Co, Ora-Gun, Gunma, Japan) at constant temperatures (-5, -8, -12, and -16 C) and
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monitored by type T thermocouples (CR10X,Campbell Scientific, Leicestershire, UK).

Temperature range of experiments was slightly higher than the reference for ideal frozen
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storage (-18 °C), according to ASLT (Accelerated Shelf Life Testing) principles (detailed by

Taoukis and Labuza, 1997). Following this methodology, samples were obtained at

appropriate time intervals for each storage temperature and a rough estimate of an expected

temperature-dependence of vitamin C loss and colour change rate from a previously

published work (Dermesonlouoglou et al., 2004).

Vitamin C determination

Page 8 of 43
 Vitamin C was determined by a high performance liquid chromatography method

(HPLC) (Giannakourou et al., 2003). All analyses were carried out in duplicate on the fruit

tissue, homogenized, using a pestle and mortar. 5 g of homogenate were mechanically stirred

in 15 ml of a 4.5 % (wt/vol) solution of metaphosphoric acid (Merck, Darmstadt, Germany)

for 15 min. The mixture was vacuum-filtered and diluted with HPLC grade water (Merck,

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Darmstadt, Germany); the total final volume was measured and an aliquot was filtered

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through a 0.45 μm syringe filter (PVDF, 0.45 μm/25 mm, Macherey-Nagel, GmbH & Co.

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KG, Düren, Germany) prior to injection into the chromatographic column. The

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instrumentation details were: HP Series 1100 (quarternary pump, vacuum degasser, a Rheo

dyne 20-μl injection loop and a Diode-Array Detector, controlled by HP Chem Station
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software); Hypersil ODS column (2504.6 mm) of particle size 5 μm; mobile phase: HPLC

grade water with metaphosphoric acid to pH 2.2; detection at 245 nm; calibrated by an
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external standard method.

Colour measurement
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A continuous objective instrumental quantification of the colour change was done

based on a measurement of CIELab values (CIE, 1978) with a CR-200 Minolta

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Chromameter (Minolta Co., Chuo-Ku, Osaka, Japan). A standard white plate (Calibration
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plate CR-200) was used to standardize the instrument under a “C” illuminant condition

according to the CIE (Commission International de l’ Eclairage). At pre-determined times of

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storage, according to the experimental design measurements they were conducted on the

same numbered strawberries (8-10 strawberries, duplicates), representative of the group, in

order to obtain consistent information.

Drip loss measurement

To estimate drip loss during thawing, frozen strawberry samples were each put on a

sieve to allow their exuded water to drip off, and then defrost for 60 min at room temperature.

Page 9 of 43
Samples were weighed at 10 min intervals. The difference between the weight at any time t

and the initial weight referred to as the initial value was defined as the drip loss of the sample

(Talens et al. 2001).

Texture measurement

Texture measurements of thawed samples were conducted by means of a texture

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analyzer (TA-XT2i of Stable Micro Systems, Surrey, England), and a TPA (Texture Profile

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Analysis) test was carried out using 8-10 strawberries (2 replicates) thawed at room

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temperature for 30 min. The test was performed on a non-lubricated flat platform using a

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knife probe with which samples were cut at a fixed rate and depth (1/3 of the initial). Texture

characteristics such as firmness, adhesiveness, elasticity, cohesiveness, chewiness and

gumminess were calculated.

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Sensory analysis
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Sensory evaluation was carried out by a panel of eight to ten trained assessors of the

accredited (according to ISO 17025) sensory laboratory of NTUA. The assessors developed a
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list of profiling attributes and agreed on the following attributes: for red colour appearance;

for texture firmness and juiciness, for flavor sweetness, sourness and aftertaste. The intensity
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of all the different sensory impressions was evaluated using a scale from 1 to 9 (1=low
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intensity and 9=high intensity). The panel also performed a preference/acceptance test using a

9-point hedonic scale (1= extremely dislike to 9= extremely like).

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2.6. Data analysis

The results for measured quality indices of all different groups of pre-treated or

untreated strawberry samples were plotted vs time for all temperatures studied. Based on the

selected kinetic modelling of each quality index, the rates of deterioration k were calculated;

Page 10 of 43
the temperature dependence of quality deterioration was then modelled by the Arrhenius

equation (described by the activation energy value EA) (Taoukis and Labuza, 1997).

2.7. Statistical analysis

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Analysis of variance (ANOVA) and Duncan multiple range tests (a=0.05) were used

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to determine statistically significant differences (STATISTICA7.0, StatSoft, Inc., Tulsa-OK,

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USA) between the different osmotic pretreatments.

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3. Results and Discussion
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3.1. Vitamin C degradation
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The results of Vitamin C content, obtained for strawberry samples were plotted versus
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time for all temperatures studied, and the apparent order of vitamin C loss (or L-ascorbic acid

oxidation) was determined. The average retention of vitamin C is expressed relative to an

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initial, average value on day 0 of the experiment (Fig. 1), where CVit represents the
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concentration of vitamin C in 100 g of raw material. In all cases, Vitamin C loss was found to

be adequately described by an apparent first order reaction (Eq. 1):

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Cvit / Cvit o *100  e kvitt (Equation 1)

where CVit and CVito are the concentrations of vitamin C at time t and zero,

respectively, and kvit is the apparent reaction rate of Vitamin C loss. Cvito for frozen

strawberry was measured to be 29.255.18 mg/100 g (average valuestandard deviation,

number of replicates= 48). According to Equation 1, values for apparent reaction rates of

Vitamin C loss, kvit, were calculated (Table 1). After the freezing process, during the

Page 11 of 43
subsequent isothermal frozen storage, strawberries exhibited a significant loss of Vitamin C

at all temperatures studied (Fig. 1a). Compared to other published work, the vitamin C

content for frozen strawberries in this study showed a better retention. For example, Mallet

(1993) measured no retention of vitamin C (100% loss) after 30 days of storage time at –5°C.

In our case, 37.6% retention was observed at the same time. The estimated rates of vitamin C

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loss kvit were lower for the osmotically pretreated samples (OPrT) (Table 1). For all series of

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samples, it can be deduced that below –12°C, Vitamin C is remarkably stable in the modified

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matrix of pre-treated samples. For example, the values of kvit for osmo-dehydrofrozen

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strawberries with glucose or HDM at storage temperature -16°C was approximately 3 times

less than the kvit values for conventionally frozen strawberries (Table 1). The respective kvit
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values for osmo-dehydrofrozen strawberries with oligofructose at -16°C were lower; 3 times

less than kvit values for osmo-dehydrofrozen strawberries treated with the conventional
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glucose as well as the non-conventional HDM. The equivalent effect of three different OPrT

treatments (Table 1) was sustained by the Duncan HSD test conducted (a= 0.05), where the
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corresponding P value was estimated to be above 0.001.

The temperature dependence of the deterioration rate kvit was then modelled by the
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Arrhenius equation (Eq. 2):

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 E  1 1 
k vit  k vit,ref exp A   
 R  T Tref  (Equation 2)
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where kvit,ref is the reaction rate of the L-ascorbic acid loss at a reference temperature

Tref, EA is the activation energy of the chemical reaction, and R is the universal gas constant.

By linearly correlating lnkvit vs 1/Tref-1/T, the EA of L-ascorbic acid loss was estimated from

the slope of the fitted line. In Figures 1a, 1b, 1c and 1d, the L-ascorbic acid loss for untreated

and osmotically pre-treated (OPrT) - with glucose, HDM and oligofructose frozen

strawberries at all temperatures studied (-5, -8, -12 and -16 C) were presented, respectively.

Page 12 of 43
The estimated activation energies, EA, for the whole temperature range studied (from -5 to -

16 °C) are 123.9±8.1, 125.2±11.7, 152.2±5.6 and 182.8±32.4 kJ/mol, for the case of

untreated (R2= 0.9915), and osmotically pre-treated with glucose (R2= 0.9828), HDM (R2=

0.9960), and oligofructose (R2= 0.9384) frozen strawberries, respectively. The calculated

values for the vitamin C loss reaction rate at the reference temperature (Tref) of -18°C were,

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0.0030±0.0004, 0.0013±0.0002, 0.0008±0.0001 and 0.0004±0.0002 days-1, for untreated, and

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osmotically pre-treated with glucose, HDM, and oligofructose, respectively. The osmotically

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pre-treated strawberry samples presented higher ΕΑ values than the one for untreated samples.

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Figure 2 depicts within the corresponding Arrhenius plot the rates of L-ascorbic loss for

untreated and OPr-oligofructose samples in the whole temperature range of study. A closer
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observation for OPr-oligofructose samples shows that at temperatures below -12⁰C a drastic
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change of the dependence on temperature occurs, that could be depicted by a "break" in the
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Arrhenius plot. This observation could be due to the glass to rubber transition range
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approaching, leading to a higher stability than predicted by the Arrhenius. This hypothesis

assumes a significant increase of the glass transition temperature (Tg´) of the osmotically
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pretreated with oligofructose strawberry tissue, which is in agreement with previous works

(Dermesonlouoglou et al., 2005; Giannakourou and Taoukis, 2003; Giannakourou et al.,

2003b), where Tg´ measurements of osmotic solutions and osmosed tissue with DSC were

conducted. To support the afore mentioned hypothesis, experimental data at a temperature

near the glass transition temperature (Tg´), thus much lower than -12 °C would be needed;

however this experiment would require extremely long measurement times.

Page 13 of 43
 The kinetic approach that is given above for vitamin C loss shows the extent of

quality deterioration and allows for the calculation of the remaining shelf life of frozen

strawberry based on a pre-defined acceptability limit. In case of vitamin C, the limit can be

set at the level of 30% loss or 70% retention (Taoukis and Labuza, 1997). Based on this level,

the shelf life of strawberries, untreated, and osmo-dehydrated with glucose, HDM and

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oligofructose, was calculated at different frozen storage conditions (Table 2), using the

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Arrhenius equation for the temperature dependence (Eq.2). According to values in Table 2,

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the osmodehydrofrozen strawberry halves can increase significantly the shelf life of the

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product at all temperatures. At the reference temperature of -18 °C often required for frozen

vegetables, the shelf life of the osmodehydrofrozen strawberry halves, osmo-dehydrated with
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glucose, HDM, and oligofructose is estimated to be 277, 423, and 821 days; when the

respective shelf life for conventionally frozen strawberry halves is no more than 120 days.
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3.2. Colour loss

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Colour degradation also has a significant effect on the quality deterioration of

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strawberries. Decrease in red (CIE a) and yellow (CIE b) intensity in frozen strawberry
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colour was the characteristic colour change during frozen storage. The organoleptically

perceived colour change described above was found to be adequately modelled by a first-
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order reaction:

DCmax  DC (Equation 3)
 e  k col t
DC max  DCO

where DC (  a  a0   b  b0  ) is the chroma change, DCmax is the asymptotic value of DC

2 2

observed after a period of storage, DCo at zero time, ao and bo the values of a and b colour

parameters at zero time, kcol is the colour change rate at temperature T. Chroma change for

Page 14 of 43
pre-treated with glucose, HDM, and oligofructose strawberry samples at all temperatures

studied is representatively shown in Figures 3, respectively. Colour measurements indicated

that colour change of untreated frozen strawberry samples was more intense than the change

of colour of pre-treated samples. Indeed, the degradation rates (kcol) of Eq. 3 for the pre-

treated strawberry samples were lower than the corresponding rates for the untreated,

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showing the protection provided by the application of the osmotic procedure prior to freezing

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(values for apparent reaction rates of colour change kcol, are given in Table 1). Statistical

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analysis (ANOVA), as well as the Duncan HSD test (a=0.05), that were conducted to assess

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the impact of different storage temperature and treatment on colour degradation rate, showed

significant differences between untreated, and pre-treated strawberries (P-value>0.001).

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Temperature dependence of the colour change rates was adequately described by Arrhenius

kinetics in the whole temperature range. The values of the activation energies were calculated
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as kJ/mol: 152.6±4.9 (R2= 0.9969), 146.0±16.4 (R2= 0.9631), 181.8±6.8 (R2= 0.9960) and

162.2±35.6 (R2= 0.9384) for untreated and pre-treated with glucose, HDM and oligofructose
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syrup strawberry samples, respectively. The values for reaction rates at the reference

temperature -18 °C were 0.0018±0.0001, 0.00097±0.00022, 0.00064±0.00006 and

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0.00072±0.00031 days-1,for untreated and pre-treated with glucose, HDM and oligofructose
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syrup strawberry samples, respectively.

The remaining shelf life for frozen strawberry halves can be found based on a pre-
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defined acceptability limit for the colour change using the same kinetic approach that has

been analysed above for vitamin C retention. The limit was set at the level of 50% “chroma”

retention (expressed by “(DCmax−DC)/ (DCmax−DCo)” parameter) where the product was

judged of unacceptable colour by the sensory panel. Based on this level, the shelf life of

strawberries, untreated, and osmo-dehydrated with glucose, HDM, and oligofructose was

calculated (Eq. 2, and 3) (Table 2). As it is clearly shown, colour was well retained in

Page 15 of 43
osmotically pre-treated samples, when compared to untreated ones. Furthermore, the use of

non-conventional carbohydrates extended products' shelf life at very low freezing

temperatures (<-12 °C) as well as at higher temperatures (>-12 °C) (Table 2), proving the

improved quality retention obtained when using alternative agents in the osmotic solution.

Assessing the most appropriate, 'representative' index to calculate frozen strawberry's shelf

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life, colour change (rather than vitamin C degradation) could be recommended. According to

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consumers, colour indicates freshness and food high quality, thus being a critical quality

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parameter particularly in fresh or processed strawberries (Bodelón et al., 2013).

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Unfortunately, the attractive brilliant red is not retained after processing and storage, and

processed strawberries are prone to rapid pigment degradation and colour loss (Bodelon et
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al., 2013; Holzwarth et al., 2013a, b). Changes in anthocyanins seem to be one of the causes

for fruit browning, and their degradation is the result of coupled oxidation by PPO in the
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presence of other phenolic compounds. By using osmotic pre-treatment with the alternative

osmotic solute, oligofructose strawberry shelf life was increased up to 3.5 times. However,
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vitamin C is an indirect attribute, which, under circumstances (e.g. nutritional labelling) can

restrict product's shelf life. Instrumental colour as well as vitamin C can be used as reliable
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and easily measured indices that reflect the whole temperature history of the product.
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Validated kinetic models can be used for evaluation, control and proper management of the

frozen chain. Sensorial evaluation of colour change was in good agreement with the
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instrumentally measured colour change.

3.3. Texture measurement

Texture measurements conducted did not allow the deduction of a consistent modelable

conclusion. Parameters such as firmness, chewiness, and gumminess of all osmotically pre-

Page 16 of 43
treated samples were higher than the untreated one, when being measured thawed.

Cohesiveness and elasticity did not differ significantly between treatments (data not shown).

Firmness values (the maximum force occurring during compression) of all osmotically pre-

treated samples were significantly higher than the respective values for the untreated one

when measured after thawing, at all temperatures studied (F>Fcrit) (strawberries stored at 0

t
days). This indicated a protective effect of the osmotic dehydration on cell integrity during

ip
frozen storage. A decrease of the maximum force values after freezing shows damaged fruit

cr
tissues, partly due to histological damage due to formation of ice crystals during freezing.

us
This increase in maximum force may be attributed to the effect of sugar uptake. This agrees

with the report stating that the presence of sugars increased thawed apple samples firmness
an
(Tregunno and Goff, 1996). Monsalve-Gonzalez et al., (1993) reported that firmness

correlated significantly with sugar uptake; Torreggiani and Bertolo (2000) stated that cells
M
protected by sugars exhibited less damage to the middle lamella and less severe shrinking of

the cell content. Calcium chloride also contributed to an increased firmness due to the
ed

interaction of calcium with cellular wall components of the plant cellular matrix (Gras et al.,

2003; Castello et al., 2010). After storage of the samples at all temperatures, the firmness
pt

values were reduced. The change was more pronounced for the osmodehydrofrozen samples
ce

(with all carbohydrates used) however the values were still higher than the corresponding

values of the untreated ones. In Figures 4a, the firmness values for strawberry samples,
Ac

untreated and pre-treated with the osmotic solutes, stored at –16 °C (180 days) are

representatively demonstrated. Strawberry samples pre-treated with HDM were more stable

during storage at -16 °C; the decrease was more pronounced for pre-treated samples with

glucose (starting from time zero). The differences between pre-treated samples were not

significant as storage time increased (>180 days).

Page 17 of 43
3.4. Drip loss

Based on results of weight reduction during thawing, all osmotically pre-treated

strawberry samples showed a better retention of their water content. This superior behaviour

is more pronounced when alternative carbohydrates are used, at all temperatures presented. In

t
Figure 4b, drip loss measurement results are demonstrated for strawberry samples, untreated

ip
and pre-treated with the osmotic solutes, stored at –16 °C (180 days). The increasing trend of

cr
drip loss measurement results were in accordance with the decreasing trend of texture

us
measurement (firmness) results. Frozen strawberry samples pre-treated with oligofructose

and HDM were stable (especially at -16°C); pre-treated samples by using glucose presented
an
less stability as shown in Figures (initial values for drip loss for samples of -16°C, untreated,

pre-treated with glucose, HDM, and oligofructose: 14.82, 5.42, 3.62 and 5.79 g/100 g,
M
respectively).
ed

3.5. Sensory evaluation

pt

The organoleptic quality of strawberry samples can be described in terms of

ce

appearance, shape preservation, exudation of sap, colour of fruit, firmness, juiciness,

aftertaste, flavour, sweetness and sourness. In Table 3, average scores for the intensity (scale
Ac

1-9) for red colour, texture (force required to cut with the front teeth), taste and overall

acceptance are shown for samples, untreated and pre-treated with all osmotic solutes used (a)

at the beginning of their storage at –16 °C (1st day) and (b) after 180 days of storage at –16

°C. The descriptive test of the strawberry samples showed significant effects of the osmotic

pre-treatment. Based on the results, untreated frozen samples suffered from a detrimental

texture and taste deterioration during their storage. Sensory evaluation also showed good

Page 18 of 43
organoleptic quality in osmodehydrofrozen strawberry halves stored for up to 6 months at –

16°C. The average scores for overall acceptance of osmodehydrofrozen strawberries with the

osmotic solutes HDM and oligofructose stored at temperature –16°C for 6 months’ time were

7.3 and 7.5, respectively, compared to scores 6.0 and 6.0 for untreated samples as well as for

osmodehydrofrozen samples with glucose, respectively, showing the improved sensory

t
attributes resulting from alternative agents application. The tissue integrity was well retained

ip
on osmotically pre-treated samples, especially when oligofructose was used in the osmotic

cr
solution. Taste of the pre-treated was judged as “pleasant and acceptable” despite differing

us
from the fresh fruit taste (sweet, non strawberry-like taste). However, the pre-treated samples

were preferred in all attributes including taste (even in the case of non-conventional
an
carbohydrates) and they showed an increased stability with storage. Statistical analysis did

not show significant differences of different quality attributes between osmotic treatments
M
with HDM and oligofructose during storage.
ed

4. Conclusions
pt

The objective was to evaluate the effect of the pre-treatment with non conventional
ce

osmotic agents (oligofructose and HDM) on the quality of delicate strawberry tissue during

frozen storage and after thawing. The osmotic dehydration prior to freezing on strawberries
Ac

has demonstrated to be useful for limiting quality loss of frozen fruit tissue. The sugar

enrichment being obtained and partial water removal improves the sensory characteristics of

treated strawberry halves, when being compared to untreated ones, which suffered from a

detrimental texture and taste deterioration during storage. The treated samples can be used in

various food applications, such as yogurt, breakfast cereals, confectionery, leading to

significant stabilization of the enriched products. Vitamin C and colour, chosen as a primary

Page 19 of 43
quality index for frozen strawberries, showed a significantly increased retention in

dehydrofrozen samples, compared to the untreated, conventionally frozen ones, in the whole

temperature range studied, including the most detrimental –5 to –16°C freezing zone, that not

infrequently occurs in the problematic chill chain. Additionally, when colour and other

sensory attributes are concerned, the use of non-conventional carbohydrates (oligofructose

t
and HDM) showed an even better protection when compared to conventional agents' use

ip
(glucose). The results of the kinetic study in the different systems, untreated or osmotically

cr
pre-treated, indicate that both vitamin C loss and colour change can be used for shelf life

us
modelling and predictions. Besides questions of theoretical validity of the Arrhenius equation

in specific frozen ranges of interest, cautious application even on empirical basis serves as a
an
useful practical tool for shelf life calculations and predictions.
M
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Page 28 of 43
Captions to Figures

Fig. 1 Vitamin C loss [expressed as (Cvit/Cvito*100) vs storage time] for (a) untreated, and

osmotically pre-treated (OPrT) with (b) glucose, (c) HDM and (d) oligofructose (osmotic

treatment conditions: 150 min at 35C) frozen strawberries, at temperatures studied: -5, -8, -

t
12, -16C (average values ± standard deviation).

ip
Fig. 2 Temperature dependence of vitamin C loss rates [Arrhenius plot: lnCvit vs (1/T)-

cr
(1/Tref=-18°C)] for untreated ( , Solid line: Arrhenius fit for temperatures from -5 to -16°C),

us
and osmotically pre-treated (OPrT) with oligofructose ( , Dashed line: Arrhenius fit for

temperatures from -5 to -16°C; Solid line: Arrhenius fit, broken, for temperatures from -5 to -

12°C, and from -12 to -16°C) frozen strawberries.

an
Fig. 3 Colour change [expressed by (DCmax-DC)/(DCmax-DCo) parameter vs storage time] for
M
(a) untreated, and osmotically pre-treated (OPrT) with (b) glucose, (c) HDM, and (d)

oligofructose (osmotic treatment conditions: 150 min at 35C) frozen strawberries, at

ed

temperatures studied: -5, -8, -12, -16C (average values ± standard deviation).
pt

Fig. 4 (a) Firmness values (Fmax, N vs storage time) and (b) drip loss after thawing at room

temperature (thawing time, 60 min) (g/100g vs storage time) of thawed strawberry samples,
ce

fresh, untreated and osmotically pre-treated with glucose, HDM, and oligofructose (osmotic

treatment conditions: 150 min at 35C) stored at –16 °C (average values ± standard
Ac

deviation).

Page 29 of 43
Table 1
Rates of Vitamin C loss (kvit, 1/d) and colour change loss (kcol, 1/d) for untreated, and
osmotically pre-treated (OPrT) with glucose, HDM, and oligofructose (osmotic treatment
conditions: 150 min at 35C), frozen strawberries, at temperatures studied (-5, -8, -12, -
16C).
Sample Storage temperature (°C)

t
-5 -8 -12 -16

ip
Rates of vitamin C loss C kvit (1/d)* (R2)

cr
_ 0.0477±0.0016 0.0303±0.0012 0.0103±0.0004 0.0048±0.0003

us
(0.9949) (0.9910) (0.9847) (0.9437)

OPrT, 0.0206±0.0008 0.0122±0.0005 0.0060±0.0003 0.0018±0.00001

glucose (0.9896) (0.9759) an (0.9697) (0.932)

OPrT, 0.0263±0.0014 0.0137±0.0012 0.0041±0.0003 0.0015±0.00001

M
HDM (0.9808) (0.9105) (0.9459) (0.9384)

OPrT, 0.0221±0.0008 0.0120±0.0009 0.0053±0.0002 0.0006±0.00004

ed

oligofructose (0.9907) (0.9212) (0.9864) (0.9159)

Rates of colour change C kcol (1/d)* (R2)

pt

_ 0.0622±0.0026 0.0250±0.0007 0.0094±0.0003 0.0032±0.00008

ce

(0.9938) (0.9948) (0.9900) (0.9928)

OPrT, 0.0248±0.0010 0.0129±0.0004 0.0071±0.0001 0.0014±0.00004

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glucose (0.9894) (0.9890) (0.9951) (0.9861)

OPrT, 0.0420±0.0022 0.0147±0.0003 0.0050±0.0002 0.0009±0.00004

HDM (0.9842) (0.9951) (0.9855) (0.9722)

OPrT, 0.0236±0.0015 0.0127±0.0003 0.0075±0.0001 0.0012±0.00004

oligofructose (0.9749) (0.9925) (0.9970) (0.9827)

* Values are mean ± standard deviation of prediction.

Page 30 of 43
 Table 2

Shelf life (days) of frozen strawberries, untreated, and osmotically pre-treated (OPrT) with

glucose, HDM, and oligofructose (osmotic treatment conditions: 150 min at 35C), frozen

strawberries based on vitamin C loss 30% or instrumentally measured colour change DCmax-

DC/DCmax-DCo= 0,50, at storage temperatures -5, -8, -12, -16°C.

t
Storage Shelf life, d Shelf life, d

ip
temperature Acceptance limit Acceptance limit

cr
(°C) for vitamin C content for colour

us
vitamin C loss= 30% DCmax-DC/DCmax-DCo= 0,50

_ OPrT, OPrT, OPrT, _ OPrT, OPrT, OPrT,

gl. HDM olig.

an gl. HDM olig.

-5 7 17 13 16 11 27 16 29
M
-8 11 29 26 29 27 53 47 54

-12 34 59 86 67 73 111 138 92

ed

-16 74 198 237 594 216 495 577 770

pt
ce
Ac

Page 31 of 43
 Table 3

Results of sensory evaluation of fresh (before freezing) and thawed strawberry samples,

untreated, and pre-treated (OPrT) with glucose, HDM, and oligofructose (osmotic treatment

conditions: 150 min at 35C), stored for 1 and 180 days at temperature –16°C, on a 9-score

t
hedonic scale.

ip
Quality Before After freezing Frozen storage

cr
factor freezing 1st day at -16C 180 days at -16C
Osmotic Pre-Treatment Osmotic Pre-Treatment

us
_ OPr OPrT, OPrT, _ OPr OPrT, OPrT,

T, gl. HDM olig.

an T, gl. HDM olig.

Colour 8.5a 7.5 a 7.5 a 7.5 a 7.5 a 6.0 a 5.5 a 7.0 a 7.0 a
M
Texture 9.0b 7.5 a 7.0 b 8.5 b 8.0 b 5.0 b 5.0 b 7.5 b 7.5 b

Taste 9.0b 7.5 a 7.0 b 8.0 c 8.5 c 6.5 c 6.5 c 7.0 a 7.0 a
ed

Overall 9.0b 7.5 a 7.0 b 8.0 c 8.0 b 6.0 a 6.0 d 7.5 b 7.3c
acceptance
* Values of means bearing different letters are significantly different (P<0.05).
pt
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