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Journal of Food Engineering 83 (2007) 41–46

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Thermal and pulsed electric fields pasteurization of apple juice:


Effects on physicochemical properties and flavour compounds
S.F. Aguilar-Rosas a, M.L. Ballinas-Casarrubias a, G.V. Nevarez-Moorillon a,
O. Martin-Belloso b, E. Ortega-Rivas a,*
a
Food and Chemical Engineering Programme, Autonomous University of Chihuahua, University Campus I, Chihuahua, Chih. 31170, Mexico
b
Department of Food Science and Technology, University of Lleida, Av. Alcalde Rovira Roure 177, 25198 Lleida, Spain

Received 6 September 2006; received in revised form 14 December 2006; accepted 15 December 2006
Available online 11 January 2007

Abstract

Apple juice, extracted from golden delicious fruits, was pasteurized using a pulsed electric field (PEF) treatment and compared with a
conventional high temperature-short time (HTST) method. The PEF treatment was carried out using a PEF laboratory unit, set with a
bipolar pulse (4 ls wide), an intensity of 35 kV/cm, and a frequency of 1200 pulses per second (pps). The thermal pasteurization was
performed at 90 °C for 30 s with an adapted laboratory set-up. Effects of variables of both treatments on pH, total acidity, phenolics
content, and volatile compounds were investigated. While minimal variability was observed in pH and no significant changes were
detected in acidity, phenolics content and volatile compounds concentration showed statistical significant differences between treatments.
In general, these measured variables were less affected by the PEF treatment than by the thermal pasteurization.
Ó 2007 Elsevier Ltd. All rights reserved.

Keywords: Apple juice; Thermal pasteurization; High voltage pulsed electric fields (PEF); High temperature-short time (HTST) pasteurization; Sensory
attributes

1. Introduction commonly used method for heat treatment of apple juice.


In HTST pasteurization, the temperature used is 76.6–
Apple juice has been traditionally pasteurized by ther- 87.7 °C for a holding time between 25 and 30 s (Moyer &
mal means. Both batch and continuous methods are used Aitken, 1980).
in apple juice pasteurization and the treatment may be car- Thermal pasteurization is quite efficient in preventing
ried out before or after packing the product in the con- microbial spoilage of apple juice but the applied heat
tainer. In batch pasteurization, individual volumes are may also cause undesirable biochemical and nutritious
treated in jacketed stainless steel vessels. The jacket may changes which may affect overall quality of the final prod-
be used both for heating (with steam or hot water) and uct. Alternative methods of pasteurization that do not
cooling (with chilled water or brine). Continuous pasteuri- include direct heat have been investigated in order to
zation may be carried out by passing the juice through obtain a product safe for consumption, but with sensory
plate heat exchangers, which usually comprise the stages attributes similar to the untreated juice. High voltage
of pre-heating, heating, holding and cooling. Currently, pulsed electric fields (PEF) treatment is a promising non-
high temperature-short time (HTST) pasteurization is a thermal processing method that may radically change
liquid food preservation technology. Treating liquid foods
with PEF may inactivate micro-organisms and enzymes
*
Corresponding author. Tel./fax: +52 614 4241868. with only a small increase in temperature, simultaneously
E-mail address: eortegar@uach.mx (E. Ortega-Rivas). providing consumers with safe, nutritious, and fresh-like

0260-8774/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2006.12.011
42 S.F. Aguilar-Rosas et al. / Journal of Food Engineering 83 (2007) 41–46

quality foods. PEF treatment is conducted at ambient tem- looking at effects on physicochemical properties and some
perature for a short time (in microseconds), and energy lost sensory attributes, with results also being promising. For
due to heating of foods is minimized (Jeyamkondan, Jayas, example, Evrendilek et al. (2000) reported no apparent
& Holley, 1999). changes in physical and chemical properties directly caused
In terms of microbial safety and energy efficiency, a by PEF treatment in apple juice and cider, while Barbosa-
study of PEF inactivation demonstrated that, for achieving Canovas, Pothakamury, Palou, and Swanson (1998) found
a seven log reduction in survivability of Saccharomyces that pH and vitamin C concentration were not significantly
cerevisiae in apple juice, PEF utilized less than 10% of affected by PEF treatment of fresh apple juice and apple
the electric energy for heat treatment (Qin, Zhang, Barb- juice reconstituted from concentrate. There is, however, a
osa-Cánovas, Swanson, & Pedrow, 1994). It has also been dearth of information in the literature related to actual
reported (Mittal, 1998) that a PEF low energy pulser with effects of PEF on composition of volatile chemical com-
an instant-charge-reversal pulse waveform was successfully pounds responsible for odour and flavour of apple juice.
used in apple cider treatment. The consumed energy was as There are neither many direct comparisons of PEF and
low as 5.76 J/ml at 20 °C, compared with the 50 kJ/kg nor- HTST treatments, in terms of quality attributes in general.
mally required in conventional thermal processing. Micro- This paper presents an investigation of a direct comparison
bial inactivation, coupled with quality retention, has also of PEF and HTST in pasteurization of apple juice, focused
been reported for apple juice pasteurization using non- on retention of volatile compounds, which have been iden-
thermal methods of preservation (Ortega-Rivas, Zárate- tified as responsible for its characteristic aroma and tasteful
Rodrı́guez, & Barbosa-Cánovas, 1998). A comparison of flavour.
ultrafiltration (UF) and PEF in apple juice pasteurization
reported six log reductions in survivability of total aerobic 2. Materials and methods
micro-organisms using the indigenous flora of the juice
(Ortega-Rivas et al., 1998). In terms of quality aspects, sol- Freshly squeezed apple juice, from golden delicious
uble solids, pH and acidity were reported practically apple variety, was extracted with a domestic juice extrac-
impaired by both techniques. Colour, however, suffered tor. The juice was pre-filtered across a bag filter and stored
changes such as browning for UF and fading for PEF (Ort- at 4 °C prior to treatment.
ega-Rivas et al., 1998; Zárate-Rodrı́guez, Ortega-Rivas, & For conventional heat treatment, an experimental set-up
Barbosa-Cánovas, 2000). was constructed (Fig. 1). As can be observed, it consisted of
Flavour components in apple juice are numerous, and sanitary containers to hold heating and cooling fluids, coils
flavour identification is considered quite complex due to for juice passage, a centrifugal sanitary pump to circulate
the aromatic nature of apples. Eight odour-active volatiles the juice, and thermocouples to record the temperature.
have been, however, identified as the most important con- A pasteurization temperature of 90 °C was tested for a
tributors for the aroma–flavour authenticity of apple juice holding time of 30 s, which was virtually the maximum
(Cunningham, Acree, Barnard, Butts, & Braell, 1986). range suggested in the literature (Moyer & Aitken, 1980).
Apparently, there are not reported studies of PEF effects Also, it was sufficient to achieve pasteurization conditions
on volatile compounds in apple juice. Several reports have using Lactobacillus brevis and S. cerevisiae, common spoil-
appeared for orange juice (Jia, Zhang, & Min, 1999; Yeom, age micro-organisms in apple juice, as contaminating spe-
Streaker, Zhang, & Min, 2000) focusing on effects of PEF cies. As shown in Fig. 2, inoculates of L. brevis and S.
on quality aspects. The PEF-treated juice was compared cerevisiae, expressed in colony forming units per millilitre
with juice pasteurized by heat at 94.6 °C for 30 s. The juice (cfu/ml) were properly reduced.
treated by PEF retained greater amounts of vitamin C and A high voltage pulsed electric field unit, designed and
some representative flavour compounds, than the juice pas- constructed at Ohio State University (Columbus, OH,
teurized by heat during storage at 4 °C. In terms of specific USA) was used for the PEF treatment. As shown in
flavour compounds, it was found that 40% of decanal was Fig. 3, this test apparatus consists of a high voltage power
lost by heat treatment at 90 °C for 3 min while no loss was
observed by PEF treatment at 30 kV/cm, either at 240 or
480 ls (Jia et al., 1999). Octanal showed a loss of 9.9% Feed

for the heat treatment and 0% for any of the two PEF Product
treatments. Some compounds suffered losses for the PEF
treatments, but always in less proportion than the heat pas-
teurized juice. For example, 5.1% and 9.7% of ethyl buty-
rate were lost for the 240 ls and 480 ls treatments,
respectively, but 22.4% was lost in the thermal process
(Jia et al., 1999).
As discussed above, PEF has been challenged against
Pre-heating Holding Cooling
many spoiling micro-organisms in apple juice, with encour-
aging results. Also, pertaining quality, there are studies Fig. 1. Experimental set-up used for heat pasteurization of apple juice.
S.F. Aguilar-Rosas et al. / Journal of Food Engineering 83 (2007) 41–46 43

9 9

8 8
L. brevis
Initial count
7 7 S. cerevisiae
Final count
6 6
log(cfu/ml)

log(cfu/ml)
5 5

4 4

3 3

2 2

1 1

0 0
L. brevis S. cerevisiae 0 400 800 1200 1600
Microbial Inactivation by HTST Method Pulses per second of PEF treatment

Fig. 2. Inactivation of Lactobacillus brevis and Saccharomyces cerevisiae Fig. 4. Inactivation of Lactobacillus brevis and Saccharomyces cerevisiae
by HTST pasteurization (90 °C for 30 s). by PEF pasteurization (pulse frequency at 35 kV/cm).

the result being expressed as g malic acid/l of sample


Treatment
High voltage High voltage
pulse chambers (AOAC, 1998).
power supply
generator Total phenol content was determined by the Folin–Cio-
HV
calteu method (Singleton & Rossi, 1965), reading samples
in a HP 845 A UV/visible spectrophotometer (Hewlett–
GND
Packard Inc., Palo Alto, CA, USA) at 760 nm. Samples
T2 T1
were centrifuged at 2000g (4 °C for 5 min) and diluted by
a factor of 10 with distilled water. Results were expressed
Product as mg of gallic acid/l of juice.
The flavour compounds in the headspace of the apple
Feed juice were analysed by solid-phase microextraction (SPME)
and gas chromatography (Buchholz & Pawliszyn, 1994).
Samples of 10 ml of apple juice were transferred into
30 ml vials. The SPME fibre coated with 100 lm poly-
Water bath
dimethylsiloxane was inserted into the headspace of the
juice and heated at 50 °C for 30 min. The SPME sample
was removed from the sample vial and inserted into a gas
Fig. 3. Diagram of pulsed electric field treatment operation. chromatograph (GC) injection port, and held for 4 min
at 250 °C to desorb the flavour compounds absorbed on
the SPME coating. The desorbed flavour compounds were
supply, a high voltage pulse generator, a series of treatment separated using an Agilent 5973 Network GC/MS equip-
chambers, and sample cooling and delivery devices. Fol- ment (Agilent Technologies, Palo Alto, CA, USA)
lowing recommendations in the literature (Heinz, Toepfl, equipped with a capillary column of 0.25 mm internal
& Knorr, 2003), a 4 ls bipolar pulse with an electric field diameter  30 m length, and coated with 0.25 lm thick
strength of 35 kV/cm was chosen to destroy the same spoil- diphenylpolysiloxane. Helium was the carrying gas at a
age micro-organisms mentioned above and select the rate of 1.5 ml/min. The GC oven temperature was pro-
appropriate frequency for further treatment. Such grammed from 40 to 250 °C at 20 °C/min and held
microbes were inoculated and inactivated using the previ- 10 min at the final temperature. At the end of the experi-
ously mentioned conditions at different repetition rates. mental run, volatiles were qualified using the library in
Using 1200 pps, 6.3 and 4.2 log reduction cycles were the program of the instrument. After identification of vol-
achieved for the bacterium and yeast, respectively atile compounds, calibration curves were derived for every
(Fig. 4), which were considered appropriate so this fre- one using the authentic volatiles. The presence of the main
quency was used for experimentation. volatiles reported to be present in apple juice were con-
The pH was measured by direct reading at 25 °C in an firmed by comparing the retention times of gas chromato-
Orion Benchtop pH/ISE-meter Model 420 A (Orion graphic peaks to those of authentic compounds.
Research Inc., Boston MA, USA). Acidity was measured The results were interpreted using simple analyses of
by titration with 0.1 N NaOH to a pH end-point of 8.2, variance (ANOVAS). For the volatile compounds, a Stu-
44 S.F. Aguilar-Rosas et al. / Journal of Food Engineering 83 (2007) 41–46

dent t-test for independent samples was used. Means were 0.355
differentiated by Tukey’s tests. Significance of differences 0.35
was defined at p < 0.05. The tests were performed in tripli-
0.345
cate. The statistic system SAS Version 8 (SAS Institute

g malic acid/litre
Inc., Cary, NC, USA) was used for actual calculations. 0.34

0.335
3. Results and discussion
0.33

An ANOVA for pH determinations showed statistical 0.325

significant difference (F = 130.40, p = 0.0001) between the 0.32


untreated apple juice and both the HTST-pasteurized and
0.315
the PEF-treated samples, as illustrated in Fig. 5. However,
by inspecting such figure, it can be observed that such dif- 0.31
Untreated PEF HTST
ferences may be considered negligible for practical pur- Treatment
poses, since the measured pH in the three samples, only
vary between 3.8 and 3.9. The observed small discrepancies Fig. 6. Effect of treatment method on acidity of pasteurized apple juice.
could be, possibly, attributed to experimental error. These
findings agree with an investigation by Heinz et al. (2003)
who reported that PEF-pasteurized apple juice did not Acidity in apple juice is an important sensory attribute
show practical difference in pH. On the other hand, associated with its characteristic flavour and astringency.
Charles-Rodrı́guez (2002) found that thermally-treated Apparently, PEF-pasteurized apple juice do not affect acid-
apple juice presented an increase in pH directly related with ity, so this important feature remains practically intact with
temperature, reaching a value of 4.01 at the extreme pas- the consequently advantage in overall quality of the
teurization conditions of 85 °C and 27 s. This author also product.
reported that PEF-pasteurized juice did not show variabil- Considering the results previously discussed, apparently,
ity in pH at different electric field strengths and frequencies. PEF maintained the physicochemical properties of the ori-
It is known that maintaining pH on low values prevent ginal juice, while HTST affected the pH and in a less extent
pathogenic microbial growth in fruit juices, so PEF treat- the acidity of the juice, as shown in Figs. 5 and 6. Apart
ment gives, apparently, more stability to pH in apple juice from the experimental error that may have caused some
than HTST pasteurization. discrepancies in readings, the changes observed in the ther-
In terms of acidity, no significant statistical difference mal method could be attributed to the evaporative effect of
was observed for any of the treatments (F = 0.94, organic acids as a function of temperature increase.
p = 0.4404), as shown in Fig. 6. These results are in agree- Contents of total phenol compounds presented variabil-
ment with the study of Heinz et al. (2003) already men- ity for the two compared pasteurization methods. The
tioned, who also reported no significant changes in ANOVA in this case indicated statistical significant differ-
acidity of PEF-pasteurized apple juice. Ortega-Rivas ence (F = 44.4, p = 0.0003) between both treatments and
et al. (1998) presented results of a comparison of UF and the control (Fig. 7). A Tukey test confirmed the difference
PEF in some physicochemical properties of apple juice, of means for the three samples. It can be observed in Fig. 7,
finding that acidity did not present significant variability. however, that the HTST treatment caused a considerable

3.94 120

3.92
100
3.9
ppm of gallic acid

3.88 80

3.86
60
pH

3.84

3.82 40
3.8
20
3.78

3.76
0
3.74 Untreated PEF HTST
Untreated PEF HTST Treatment
Treatment
Fig. 7. Effect of treatment method on total phenol compounds of
Fig. 5. Effect of treatment method on pH of pasteurized apple juice. pasteurized apple juice.
S.F. Aguilar-Rosas et al. / Journal of Food Engineering 83 (2007) 41–46 45

lost of phenols (32.2%) when compared with the PEF treat- To the extent of the literature survey of this work,
ment, which only caused a 14.49% reduction. These results there are not reported studies of PEF effects on volatiles
agree with Spanos and Wrolstad (1992), who reported that in apple juice. As previously discussed, several reports
total phenol concentration is reduced up to 50% in apple have appeared for orange juice (Jia et al., 1999; Yeom
juice pasteurized thermally at 80 °C for 15 min. Gardner, et al., 2000) focusing on effects of PEF on quality aspects.
White, McPhail, and Duthie (2000) observed also consider- Although orange juice and apple juice are different prod-
able losses in phenolics in apple juice pasteurized by ther- ucts, the studies mentioned for orange juice, along with
mal means. Phenol compounds are secondary metabolites the results of this work, may be indicative of a definite
in plants known to play an important role in colour and evaporative effect of thermal pasteurization methods in
flavour development in fruit juices and wine. Phenols are fruit juice pasteurization in general. Thus, in a very broad
important constituents of pear, grapes and apple and way, it may be considered that non-thermal pasteurization
may be categorized into two groups: phenolic acids and techniques will represent a better choice for processing of
flavonoids (Spanos & Wrolstad, 1992). The combined fruit juices in general, and apple juice in particular. Vola-
odour–flavour characteristics in apple and apple products tile chemical compounds responsible for colour and fla-
are due in part to phenol compounds. Phenols are also used vour of fruit juices are retained in a highest ratio, when
as indicators of physiological state and potential damage in compared with fresh untreated samples, by the use of
quality of fruit products (Blanco, Fraga, & Mangas, 2001). non-thermal pasteurization techniques such as high volt-
Phenol compounds are, thus, important biochemical sub- age pulsed electric fields. PEF pasteurization of apple
stances in apple juice. Their lost or decrease in concentra- juice may be, therefore, considered a feasible alternative
tion will, therefore, impair seriously apple juice sensory for fruit processors, in order to obtain a premium quality
attributes. PEF treatment with minimal losses (Fig. 7) product.
would represent an obvious advantage over HTST pasteur-
ization, in terms of concentration of these chemicals in 4. Conclusions
apple juice.
As previously stated, the flavour of apple juice consists PEF, a thermal preservation technique to pasteurize
of many chemical compounds, but the literature indicates apple juice, proved to be efficient in microbial inactivation,
8–23 compounds most responsible for the odour–flavour as well as in preserving some quality attributes. Conven-
attribute (König & Schreier, 1999; Rao, Acree, Cooley, & tional HTST pasteurization, on the other hand, produced
Ennis, 1987). Eight volatile compounds were properly iden- significant losses in phenolic compounds and in volatiles
tified in the apple juice, fresh and processed by any of the responsible for flavour. PEF-treated juice retained better
treatments, in this work. Table 1 compares the percentage most of the volatile compounds responsible for colour
in concentration decrease for those mentioned volatiles in and flavour of the apple juice. Further studies on the chem-
the pasteurization methods investigated. All volatile con- istry of flavour components, to try to preserve them even
centrations showed statistical significant differences for better, are advisable. It is also recommended to use sensory
both treatments, as compared with the untreated sample, evaluation to define quality differences of apple juice trea-
by a Student t-test for independent samples (p < 0.05, ted by non-conventional methods. The use of PEF as an
n = 3). In all compounds, and particularly in one (ethyl alternative to heat pasteurization of apple juice may be
acetate), the decrease was considerable higher for the considered a strategically important action to obtain a sen-
HTST treatment than for the PEF method (Table 1). Some sory impaired product, highly competitive in global
of the volatiles in the PEF treatment were almost retained. markets.
Hexanal and hexyl acetate were only lost in 7% and 8.4%,
respectively. It is worth to mention (Table 1) that acetic Acknowledgements
acid was completely lost in HTST treatment.
An experimental part of this project was carried out at
the Department of Food Technology of University of
Table 1 Lleida, Spain. The authors wish to express their gratitude
Percentage of volatiles losses, compared with untreated sample, in apple for the assistance provided by technical and academic
juice treated by two methods staff. Funding for the project was provided by the Na-
Compound Loss percentage for PEF Loss percentage for HTST tional Council of Science and Technology (CONACyT,
Acetic acid 39.792 ± 20.84 100 México).
Hexanal 7.042 ± 9.32 62.348 ± 5.35
Butyl hexanoate 18.108 ± 7.72 36.273 ± 24.86 References
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