The effect of high pressure on microbial population, meat quality and sensory

characteristics of chicken breast llet

Zbigniew A. Kruk
a, b
, Hyejeong Yun
a
, David L. Rutley
b
, Eun Jung Lee
c
, Yun Ji Kim
c
, Cheorun Jo
a,
*
a
Department of Animal Science and Biotechnology, Chungnam National University, Daejeon 305-764, Republic of Korea
b
Livestock Systems Alliance, The University of Adelaide, Roseworthy SA 5371, Australia
c
Division of Food Safety, Korea Food Research Institute, Songnam, 463-746, Republic of Korea
a r t i c l e i n f o
Article history:
Received 4 February 2010
Received in revised form
31 May 2010
Accepted 12 June 2010
a b s t r a c t
High hydrostatic pressure (300, 450 and 600 MPa) was used to investigate its effect on microbial pop-
ulation, meat quality and sensory characteristics of chicken breast llets. Pressures of 450 and 600 MPa
almost completely eliminated all the 3 major pathogens Salmonella typhimurium (KCTC 1925), Escherichia
coli (KCTC 1682), and Listeria monocytogenes (KCTC 3569) and therefore improved safety of chicken breast
llets. The 600 MPa treatment reduced bacteria count by 6e8 log (CFU/g) for 7e14 days, and the 450 MPa
treatment reduced bacteria count by 4e8 log (CFU/g) for 3e14 days, depending on the microorganism.
The increased pressure had an impact on avour, aroma strength and juiciness. The 300 MPa pressure
signicantly reduced avour, aroma strength and juiciness, and 450 MPa produced breast llets with the
weakest aroma. Increasing pressure increased cooking loss and colour by increasing L*, a* and b* values.
Moreover, elevated pressure increased hardness, cohesiveness, gumminess and chewiness, as well as
improved freshness of meat by reducing VBN. Pressure of 450 MPa and higher induced lipid oxidation.
The results demonstrate that high pressure treatment is an effective technology in reducing bacterial
spoilage and extending shelf-life of chicken breast llet, however it may have a negative impact on some
quality and sensory characteristics.
2010 Elsevier Ltd. All rights reserved.
1. Introduction
High pressure processing technology (HPP) was originally
developed in 1899 and has been successfully used in chemical,
ceramic, steel and plastic industries (Rastogi, Raghavaro,
Balasubramaniam, Niranjan, & Knorr, 2007). It has also been
implemented in the food industry to control bacterial populations
in food products. The capacity of HPP to eradicate microorganisms
regardless of the geometry of the product, without the formation of
detrimental heat damage changes and the use of preservatives/
additives (Zhang & Mittal, 2008), made this technology quickly
accepted as safe and consumer friendly (Rastogi et al., 2007).
However, it has been reported that HPP can impact structural,
physiochemical, morphological and textural characteristics of the
meat, and can cause partial discolouration of fresh red meat
(Cheftel, 1995; Kim, Lee, Lee, Kim, & Yamamoto, 2007). Such altered
characteristics may have an effect on consumer sensory perception
of HPP treated meat. Other research has reported that there was no
effect on sensory attributes of beef treated with 600 MPa at 20

C
(Hayman, Baxter, ORiordan, & Stewart, 2004). There is limited
information available on the sensory attributes of chicken meat
treated with HPP. Therefore the aim of this study was to investigate
the effect of varying pressure levels (300, 450 and 600 MPa) on the
microbial population, quality characteristics, and the sensory
attributes of chicken meat as judged by semi-trained consumer
taste panels.
2. Materials and methods
2.1. Sample preparation and pathogens inoculation
Commercially available chicken breast llets were purchased
from the local market (Orpum Co. Ltd., Sangju, Korea), and samples
used for the pathogen experiment were completely sterilized by
gamma irradiation (35 kGy) using a cobalt-60 gamma irradiator
(AECL, IR-79, MDS Nordion International Co. Ltd., Ottawa, ON,
Canada). The irradiator source strength was 320 kCi with a dose
rate of 20 kGy/h at 10 0.5

C. Three pathogens, Salmonella
* Corresponding author. Department of Animal Science and Biotechnology,
Chungnam National University, 220 Gung-dong, Yuseong-gu, Daejeon 305-764,
Republic of Korea. Tel.: 82 42 821 5774; fax: 82 42 825 9754.
E-mail address: cheorun@adelaide.edu.au (C. Jo).
Contents lists available at ScienceDirect
Food Control
j ournal homepage: www. el sevi er. com/ l ocat e/ f oodcont
0956-7135/$ e see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodcont.2010.06.003
Food Control 22 (2011) 6e12
Typhimurium(KCTC 1925), Escherichia coli (KCTC 1682), and Listeria
monocytogenes (KCTC 3569) obtained from the Korean collection
for type culture (KCTC, Daejeon, Korea) were cultivated at 37

C for
18 h in a tryptic soy broth (Difco, Laboratories, Sparks, MD, USA).
The activated cell cultures were centrifuged at 2795g for 10 min in
a 4

C refrigerated centrifuge (Vs-5500, Vision Scientic, Co., Seoul,
Korea). The pellet was washed twice with sterile saline (0.85%), and
re-suspended in saline to a nal cell density of approximately
10
8
e 10
9
CFU/mL. Radiation-sterilized chicken breast (10 g) was
inoculated with 0.1 ml of S. Typhimurium, E. coli, and L. mono-
cytogenes, respectively, in 5 different areas and then sealed and
incubated at 10

C for 1 h to facilitate attachment of microorgan-
isms to the chicken breast. Samples used for the sensory evaluation
were not subject to sterilization or microbial treatment.
2.2. Hydrostatic pressure treatment
The samples were transported to the Korea Food Research
Institute (Seongnam, Korea) in a cooled container. Samples were
placed in a pressure vessel submerged in hydrostatic uid medium
(Quintus food processor 6; ABB Autoclave Systems, Inc., Columbus,
OH, USA), and pressurised at 300, 450 and 600 MPa for 5 min with
the initial temperature of the pressure vessel at 15 3

C. The rate
of pressurisation was 5e7 MPa/s and the pressure in the chamber
was released within 10 s. Control samples were maintained under
atmospheric pressure at 4

C while the other samples were treated.
Immediately after treatment, all samples were transported on ice to
Chungnam National University laboratory. Samples for sensory
evaluationwere stored at 4

C until required, and the other samples
for microbial analysis were immediately prepared.
2.3. Microbial analysis
After high pressure treatment, samples were blended with
sterile saline (0.88% NaCl) using a stomacher (BagMixer 400,
Interscience Ind., St. Nom, France) for 2 min. Serial dilutions were
prepared with sterile saline. Each diluent (0.1 ml) was spread on
plates in triplicate. Tryptic soy agar (Difco Laboratories, Detroit, MI,
USA) media was used for enumeration of microorganisms. Plates
were incubated at 37

C for 48 h and microbial counts were
expressed as log CFU/g. Microbial analysis was conducted during
storage at 4

C for 14 days with detection limit at 10
2
CFU/g.
2.4. Moisture and fat content analysis
The moisture content was determined using the AOAC Ofcial
Method 950.46 (AOAC, 1990). The fat content was determined by
solvent extraction using the Soxhlet technique (AOAC, 1990). Each
extracted group was then dried in an oven at 105

C for 24 h, cooled
in a desiccator, and then weighed to obtain the dry solid content.
2.5. Instrumental colour measurement
Instrumental colour measurements of the surface of the meat
sample were performed using a Hunter colorimeter (Minolta Co. CR
300, Tokyo, Japan), and Hunter colour L
*
- (lightness), a
*
- (redness),
and b
*
- (yellowness) values were determined. The mean of 10
measurements taken at different locations was recorded for each
sample.
2.6. Volatile basic nitrogen (VBN) determination
Measurements of VBN were performed according to Conway
(1950). Samples (3 g) were homogenized for 2 min with 3 ml of
distilled water and 6 ml of 10% trichloroacetic acid. After
homogenization was complete 18 ml of 5% trichloroacetic acid was
added. The mixture was centrifuged at 3000g for 10 min at 4

C,
5% trichloroacetic acid to 30 ml in total. Boric acid (0.02 N) was used
as a VBN absorber and placed in the inner section of a Conway
micro-diffusion cell. One milliliter of the sample solution and 1 ml
of K
2
CO
3
solutionwere also placed on the outer section of a Conway
micro-diffusion cell and the mixture was stirred gently. The cell was
incubated at 37

C for 60 min and then titrated against 0.02 N
sulphuric acid. The concentration of VBN was calculated as
ammonia equivalent using the following equation: VBN (mg%)
0.28 (Yt Yb) 1000; where Yt titration volume of sample
solution and Yb titration volume of blank.
2.7. Measurement of lipid oxidation
The extent of lipid oxidation was measured using the 2-thio-
barbituric acid reactive substances (TBARS) method described by
Ahn, Olson, Jo, Love, and Jin (1999) with a few modications.
Twelve millilitres of deionised distilled water (DDW) was added to
the chicken breast llet samples (3 g), and homogenized with
50 ml of butylated hydroxyanisol 7.2% (SigmaeAldrich, Saint Louis,
MO, USA) for 15 s. The meat homogenate (1 ml) was transferred to
a disposable test tube and then 1 ml of 2-thiobarbituric acid
(SigmaeAldrich, Saint Louis, MO, USA) and trichloroacetic acid
mixture (20 mM TBA in 15% trichloroacetic acid) solution were
added. The mixture was vortexed and incubated in a boiling water
bath for 15 min. The sample was then cooled in cold water for
10 min, and centrifuged for 15 min at 2500g at 4

C. The absor-
bance was measured at 532 nm using a UV-240 spectrophotom-
eter (Shimadzu Co., Kyoto, Japan), and lipid oxidation was
expressed as mg of malonaldehyde/kg meat.
2.8. Texture prole analysis
Meat samples were cut into slices 30 mm in diameter and
20 mm in thickness and used for texture prole analysis. The core
of the sample was compressed twice to 80% of its original thickness
using a Texture Analyzer (Model TA-XT 2i, Stable Micro systems
Ltd., Surrey, UK). A round needle type probe (75 mm diameter) was
set and moved perpendicularly to the sample with a speed of
1.00 mm/s, and trigger force of 0.005 kg. Texture analysis was
automatically performed by the texture expert software (version
4,0,12,0. Stable Micro systems Ltd.), and the following parameters
were recorded: hardness (N/cm
2
) maximum force required to
compress the sample; cohesiveness extent to which sample could
be deformed prior to rupture (A2/A1, where A1 is the total energy
required for rst compression and A2 as the total energy required
for the second compression); adhesiveness work necessary to
pull the compressing plunger away from sample (negative area
under the baseline between the compression cycles); gumminess
(N/cm
2
) force necessary to disintegrate a semi-solid sample for
swallowing (a combination of hardness and cohesiveness); chew-
iness (N/cm) work to masticate the sample for swallowing
(hardness cohesiveness springiness); resilience negative
force input/positive force input during the rst compression; and
springiness (cm) ability of sample to recover its original form
after a deforming force has been removed (time duration of force
input during the second compression/time duration during the rst
compression) (Bourne, 1978).
2.9. Sensory evaluation
Two semi-trained sensory panels were used for the analysis of
chicken breast llets. One panel consisted of students (15e25 years
of age) containing approximately an equal number of males and
Z.A. Kruk et al. / Food Control 22 (2011) 6e12 7
females (15 panellists per session). The other consisted only of
females, which were housewives between 26 and 45 years of age.
Chicken breast samples were sliced into 1 cm thick portions and
grilled on both sides (George Foreman Lean Mean Fat Reducing
Grilling Machine, 2400 W) for approximately 45 s to reach an
internal temperature of 71e75

C. Immediately after grilling,
samples were provided to the sensory panel using a coded identi-
er. Before tasting, panellists were familiarized with the assess-
ment criteria, the meat attributes to be rated, and how to properly
complete the questionnaire. Each treated sample was tasted by at
least 3 different panellists. Drinking water was provided to cleanse
the mouth cavity between testing each sample. Panellists used
a 9-point hedonic scale to assess various meat quality attributes.
Sensory-textural attributes scored were: meat colour (extremely
light to extremely dark), aroma strength (very weak to very strong),
aroma pleasantness (extremely dislike to extremely pleasant),
tenderness e force required to bite the sample (extremely tough to
extremely tender), juiciness (extremely dry to extremely juicy),
texture e the experience during chewing the samples, in particular
their stickiness to the roof of the mouth (extremely gooey to
extremely smooth), avour (extremely unpleasant to extremely
enjoyable), overall satisfaction (disagreeable to enjoyable), and
lastly, Would you buy this meat? (denitely not to denitely yes).
Additionally, there was space provided for further avour
description and additional comments.
2.10. Statistical analysis
The data were analyzed using general linear models (Proc GLM,
SAS Institute, 1989). Treatment (pressure) was the only design
effect in this trial and was tested as a xed level factor, with
signicance dened at the 5% level. Where the effect of treatment
was signicant the least squares means were estimated along with
their standard errors and reported. The residual correlations were
calculated and classied as either low (0 < jrj 0.3), medium
(0.3 < jrj 0.6) or high (0.6 < jrj).
3. Results and discussion
3.1. The effect of high pressure on pathogen survival
The increase of hydrostatic pressure had an effect on all three
pathogens that were used for the inoculation of chicken breast
llets (Table 1). The most dramatic effects were observed with 450
and 600 MPa pressures which completely inactivated E. coli and
L. monocytogenes (to below detectable levels). A similar trend was
observed for Salmonella typhimurium except that there was no
difference between the control and 300 MPa treatments, and the
450 MPa pressure signicantly reduced the pathogens by 55%
however, they were still detectable (Table 1).
Pressure also effected storage time at 4

C (Table 1). Three
hundred MPa reduced E. coli at days 3 and 7, however, it was not
signicantly different from the control on day 14th. S. typhimurium
was more resistant at this pressure and was only reduced signi-
cantly at day 3 of storage. On days 7 and 14 the levels were not
different from the initial (day 0) level. No effect was observed on
L. monocytogenes. The population of this strain was reduced in
relation to non-pressurised control levels by approximately 34%
however storage time did not lead to a signicant increase L. mon-
ocytogenes population.
Four hundred fty MPa was more efcient pressure in inactiva-
tion of pathogens than the 300 MPa. At this pressure level, E. coli
spoilage was reduced below the detectable levels on days 0 and 3,
and to a very low level on day 7. The results of days 0 and 3 were
signicantly different from day 14 but the results from day 7 were
not different fromthe other days. The population of E. coli on day 14
was also 55% lower than the control. S. typhimurium was non-
detectableonday3, however at days 0, 7and14levels werethe same
and signicantly less than the control. L. monocytogenes concen-
tration at every single time point was below the detectable levels.
A similar trend for L. monocytogenes occurred at 600 MPa
pressure. The effect of 600 MPa on S. typhimurium and E. coli was
similar. Both strains were not detectable on days 3 and 7, and were
signicantly reduced on day 14 (84% and 86%, respectively)
compared to the control. The 600 MPa treatment was the most
efcient pressure for inactivating all 3 bacterial strains.
Although the resistance of microorganisms to pressure was
variable, the effect of pressure treatment on chicken breast llets
microbial populations in our study was in agreement with others
(Cheftel, 1995). The most pronounced effect on bacteria inactivation
was observed with increasing pressure (Table 1). The most effective
pressures were 450 and 600 MPa, respectively, which inactivated
all three bacterial strains to almost undetectable levels. Gola, Mutti,
Manganelli, Squarcina, and Rovere (2000) demonstrated that
increasing pressure between 400 and 700 MPa caused signicant
reductions of eight E. coli strains mixed together. Malicki, Sysak, and
Bruzewicz (2005) showed that pressure between 100 and 400 MPa
efciently reduced strains of Salmonella. Styles, Hoover, and Farkas
(1991) reported a >7-log reduction in L. monocytogenes at
approximately 340 MPa pressure and Patterson, Quinn, Simpson,
and Gilmour (1995) found a similar reduction of L. monocytogenes
at 400 MPa pressure. Pressures of 450 MPa and 600 MPa were also
very effective in increasing shelf life of chicken breast llet up to 14
days of storage at 4

C. Pressure treatment between 400 and
700 MPa was reported to increase shelf life of minced meat under
refrigeration conditions (Gola et al., 2000). The results clearly
demonstrate that the increased hydrostatic pressure was able to
inactivate microbial populations and extend the shelf life of chicken
breast llet.
3.2. The effect of high pressure on sensory attributes
High pressure processing affected avour, juiciness and aroma
strength of chicken breast llet (Fig. 1). The 300 MPa reduced the
avour pleasantness and was signicantly lower than the 450 MPa
treatment (P < 0.049). The 600 MPa pressure also reduced avour,
Table 1
Effects of high pressure processing on microbial populations (log CFU/g) of chicken
breast llet during storage at 4

C.
Pathogen High
pressure
(MPa)
Storage (day) SEM
a
0 3 7 14
E. coli KCTC 1682 0.1 8.45
w
7.98
x
7.84
x
8.01
x
0.163
300 6.76
ax
5.39
by
5.97
bx
6.88
ax
0.173
450 nd
bz
nd
bz
1.30
aby
3.62
ay
0.717
600 nd
bz
nd
bz
nd
by
1.95
az
0.125
SEM
b
0.149 0.110 0.670 0.327
S. Typhimurium
KCTC 1925
0.1 6.17
x
6.74
x
6.69
x
6.84
x
0.348
300 5.53
x
5.26
y
5.06
x
5.38
x
0.255
450 2.82
y
nd
z,c
1.48
y
1.00
y
0.738
600 nd
bz
nd
bz
nd
by
1.00
ay
0.500
SEM
b
0.230 0.224 0.763 0.543
L. monocytogenes
KCTC 3569
0.1 7.35
ax
6.08
bx
5.63
bx
6.92
ax
0.115
300 4.13
y
4.38
y
4.40
xy
4.89
y
0.314
450 nd
z
nd
z
nd
z
nd
z
e
600 nd
z
nd
z
nd
z
nd
z
e
SEM
b
0.097 0.063 0.103 0.297
Values with different letters (aec) within the same row differ signicantly
(P <0.05), values with different letters (wez) within the same column differ
signicantly (P <0.05), SEM
a
standard errors of the mean (n 16),
SEM
b
standard errors of the mean (n 16), nd
c
not detected (<2.0 log CFU/g).
Z.A. Kruk et al. / Food Control 22 (2011) 6e12 8
and was not different (P < 0.0887) to 450 MPa. Higher pressure
inuenced chicken llet aroma strength. A signicant difference
was observed between 450 MPa and 600 MPa with the 450 MPa
pressure giving the weakest aroma strength (P < 0.024). There was
no difference between the control, 300 MPa and 600 MPa treat-
ments. High pressure treatment tended to reduce juiciness with
increased pressure, however the only statistically signicant effect
was observed between the control and 300 MPa treatment, which
was lower in juiciness (P <0.044). The remaining sensory attributes
such as meat colour, texture, tenderness, aroma pleasantness and
overall satisfaction were not affected by pressure.
No evidence for deteriorating effects of high pressure treatment
on sensory quality on various meat products were observed by
Hayman et al. (2004), even if the products were treated with
600 MPa pressure. Crehana, Troya, and Buckley (2000) also
concluded that high pressure processing does not markedly alter
taste, avour or nutrient content of food. However, our results
demonstrate that pressure treatment impacted avour, juiciness
and aroma pleasantness of chicken breast llet (Fig. 1). Rivas-
Canedo, Fernandez-Garcia, and Nunez (2008) showed that
pressurisation of minced beef and chicken breast (400 MPa)
signicantly changed the levels of some volatile compounds, a few
alcohols and aldehydes were decreased whereas other compounds
were more abundant in highly processed meats. This could have an
impact on avour, especially, aroma strength as observed in this
study. High pressure treatment may also accelerate other reactions
that impact food avour. Cheah and Ledward (1996) stated that the
changes leading to catalysis of lipids oxidation in pressure pro-
cessed meat were initiated at around 300 MPa at room tempera-
ture. In our study, aroma pleasantness was signicantly lower at
450 MPa and avour less acceptable at 300 MPa pressures,
respectively (Fig. 1). The effect of pressure on juiciness has been
reported by Crehana et al. (2000), who demonstrated that the
application of 300 MPa pressure signicantly increased juiciness of
frankfurters. Our results demonstrate that pressure of 300 MPa
signicantly decreased juiciness. The discrepancy between these
results could be due to the salt content differences between the
products having an impact on juiciness characteristics (Crehana
et al., 2000). The effect of high pressure treatment on sensory
attributes of chicken meat has variable effects that are benecial in
some cases and detrimental in others. The mechanism of these
variable effects is not fully understood and requires further
research (Cheftel, 1995).
3.3. The effect of high pressure on instrumental measurements of
meat parameters
3.3.1. Moisture content, cooking loss and weight reduction
Increasing pressure elevated moisture content of the samples
(Table 2). Pressures of 300 MPa and 450 MPa increased moisture by
w3% whereas 600 MPa raised moisture by almost 5%. The increased
moisture content by hydrostatic pressure paralleled with cooking
loss. Although 300 MPa pressure did not cause signicant cooking
loss when compared to the control, samples treated with 450 MPa
and 600 MPa lost signicantly more weight after cooking (Table 2).
Lean muscles consist of approximately 75% water of which
majority is held within the structure of the muscle and muscle cells
(Huff-Lonergan & Lonergan, 2005). Therefore, any treatment
affecting structural changes in the muscle can cause the release of
water entrapped within the muscle structures. HPP has been
shown to inuence the structure and function of muscle proteins
(Gross & Jaenicke, 1994; Lee, Kim, Lee, Hong, & Yamamoto, 2007;
Weber, 1989). With increased pressure muscle bers become
ner and more compact (Kim et al., 2007). Pressurisation of beef
has been shown to reduced sarcomere length and increased
cooking loss (Jung, Ghoul, & de Lamballerie Anton, 2000). In the
semitendinosus (ST) muscles cooking loss increased gradually with
increasing pressure up to 300 MPa. However, no further differences
were observed when ST was pressurised to 300, 400 and 500 MPa
(Kim et al., 2007). In our experiment utilising chicken breast llets
the pressure of 300 MPa caused a signicant increase of moisture
content, however, the cooking loss was not signicantly different
than the control. Only a higher pressure of 450 and 600 MPa
signicantly increased cooking loss by 6.4 and 19.7%, respectively.
The structural changes induced by HPP treatment in chicken
breast llets were also most likely responsible for the weight loss of
the raw samples. Applying increasing pressure on the chicken
breast llets tended to make them lighter, and this trend reached
signicance at 600 MPa where the samples were 2.69% lighter than
the control. Sample weights at 600 MPa were also signicantly
lower than those that were treated with 300 and 450 MPa (Table 2).
As mentioned previously, increases of pressure during the HPP
treatment increased moisture content in the chicken breasts in the
present study. Taking this into account, as well as the reduction of
sample weights with increased pressure, leads us to hypothesize
that the changes in muscle architecture caused by HPP treatment,
Fig. 1. The effect of high pressure processing on avour, juiciness and aroma strength
of chicken breast llet. Note: Each column represents the mean of 15 observations;
avour, juiciness and aroma strength were assessed by sensory panel using a hedonic
(1e9) scale where the avour ranged from extremely unpleasant to extremely
enjoyable, juiciness from extremely dry to extremely juicy, and aroma strength from
very weak to very strong. Error bars represent standard deviations (n 15).
Table 2
Signicant effects of high pressure processing on instrumental measurements of
meat quality.
Parameter Control 300 MPa 450 MPa 600 MPa SEM
a
Moisture content (%) 70.3
a
72.2
b
72.7
b
73.8
c
0.46
Cooking loss (%) 31.6
a
30.4
a
33.7
b
37.8
c
0.50
L*
b
52.9
a
69.9
b
78.4
c
79.3
c
0.82
a*
c
0.4
a
2.8
b
2.2
bc
1.7
c
0.38
b*
d
10.2
a
13.4
b
12.2
c
12.4
c
0.28
VBN3
e
27.1
a
17.5
b
16.1
b
14.0
c
1.05
VBN7 66.3
a
34.3
b
25.9
c
19.6
d
1.35
Hardness (kg) 28.0
b
30.1
b
44.8
a
50.7
a
3.60
Cohesiveness (%) 0.17
b
0.25
a
0.27
a
0.32
a
0.02
Gumminess (kg) 4.9
c
7.8
bc
12.3
ab
16.2
a
1.48
Chewiness (kg*mm) 1.77
b
2.72
b
4.43
a
6.70
a
0.86
TBARS0
f
0.28
a
0.27
a
0.62
b
1.17
c
0.10
TBARS3 0.48
a
0.43
a
0.66
b
1.75
c
0.08
TBARS7 0.47
a
0.73
a
0.75
a
2.32
b
0.25
Weight difference (%) 1.84
a
1.95
a
2.14
a
2.69
b
0.18
Values with different letters (aec) within the same row differ signicantly
(P <0.05),
a
SEM standard errors of the mean,
b
L* lightness,
c
a* redness,
d
b* yellowness,
e
VBNvolatile basic nitrogen,
f
TBARS 2-thiobarbituric acid
reactive substances and the number indicate the day of the experiment they were
measured.
Z.A. Kruk et al. / Food Control 22 (2011) 6e12 9
especially the reduction of space for water to be held in the
myobrils, forced the water entrapped in the muscles into the
extramyobrillar spaces. Once there it can be more easily lost
(Kristensen & Purslow, 2001; Melody et al., 2004.). Since it has been
reported by Campus et al. (2010) that water holding capacity was
negatively correlated with high pressure, this could be the main
reason for decreasing weight of chicken raw meat samples in our
experiment after pressurisation.
3.3.2. Colour characteristics
There was a signicant effect of pressure on colour character-
istics of chicken breast llets (Table 2). With increased pressure,
samples become brighter (higher L* values), and more red (higher
a* values). Although 600 MPa pressure reduced a* values signi-
cantly when comparing with 300 and 450 MPa, the difference was
still signicantly higher when compared with the control. The
increased pressure also increased b* value (yellowness) of
samples.
It has been reported that application of high pressure treat-
ment affects the colour of beef muscle (Shigehisa, Ohmori, Saito,
Taji, & Hayashi, 1991; Serra et al, 2007), sh minced muscle of
albacore tuna (Ramirez-Suarez & Morrissey, 2006), cod and
mackerel (Ohshima, Ushio, & Koizumi, 1993), bluesh and
sheephead (Ashie & Simpson, 1996) as well as meat products such
as sausages causing their whitening effect (Crehana et al., 2000).
The discolouration of meat has been observed even at lower
pressure, between 200 and 350 MPa (Carlez, Veciana-Nogues, &
Cheftel, 1995) and is accounted for myoglobin denaturation, and/
or heme displacement or release as well as ferrous oxidation
caused by the pressure treatment (Carlez et al., 1995; Mor-Mur &
Yuste, 2003). The paleness of meat treated by HPP results in an
increased brightness as measured by L* value and it is not only
accounted for a loss of active pigment but also to protein coagu-
lation that changes samples surface properties and changes the
ratio of absorbed vs reected light creating the whitening colour
effect (Goutefongea, Rampon, Nicolas, & Dumont, 1995). It has
been reported that pressure from 200 MPa inhibits calpastatin and
further increase to over 400 MPa causes degradation of calpains
(Hugas, Garriga, & Monfort, 2002). Our results in chicken breast
llet are in agreement with the other studies and have shown that
with the increasing pressure the L* value was also increased by 32,
48, and 50% for pressures of 300, 450 and 600 MPa, respectively.
The L* value increased with the pressure of 300 MPa and reached
the maximum at pressure 450 MPa. There was no signicant
difference in L* value when pressure was increased to 600 MPa
(Table 2). A similar effect was observed in pork muscle homoge-
nates. The L* value started increasing with the pressure between
100 and 200 MPa and reached the plateau between 300 and
400 MPa with no further increase at 600 MPa.
The higher a* values of pressurised chicken breast llets in our
experiment was not in agreement with the trends observed in
chicken minced breast meat (Mariutti, Orlien, Bragagnolo, &
Skibsted, 2008) as well as in ST beef muscle (Kim et al., 2007)
where the a* value was decreased by the higher pressure. This
discrepancy between the results could be accounted for the
differences between the experiments, e.g. treatment and species.
The minced chicken breast in the experiment conducted by
Mariutti et al. (2008) was minced with sage and garlic which are
natural antioxidants. The increased antioxidant status could have
a protective effect, especially after the changes induced by the high
pressure treatment that triggered ferrous oxidation of myoglobin
and in turn, affected the colour. Beef muscles have much more
intensive red colour than chicken due to a higher myoglobin
content (Hansen, Trinderup, Hviid, Darr, & Skibsted, 2003).
Therefore, the effect of pressure, especially on redness of the
muscle could be not as well pronounced in the chicken breast llets
deriving from our experiment.
Conicting reports about the effect of high pressure on b* values
in different experiments show that this colour parameter is not
always affected by the high pressure treatment. In minced chicken
breast meat no effect of pressure on b* value was observed
regardless of the presence of spices, applied pressure or storage
time (Mariutti et al., 2008). No changes of any colour parameters
studied by Ananth, Dickson, Olson, and Murano (1998) were
affected by pressure of 414 MPa in raw loin pork. b* (yellowness)
values of Catalan thin, cured, dry pork sausage (fuet) were reduced
signicantly by pressure, whereas no changes were observed in
chorizo, a sausage made with ground pork and spicy seasonings
(Marcos, Aymerich, & Garriga, 2005). In our experiment, the pres-
sure of 300 MPa increased the b* value by 31% compare with the
control and the pressure of 450 and 600 MPa decreased the b*
values which were still signicantly higher than the control by 19
and 21%, respectively (Table 2). A slight elevation of b* value of ST
muscle pressurised to 400 and 500 MPa was observed by Kim et al.
(2007). However, the conicting results between various experi-
ments regarding a* and b* colour parameters could be also
accounted by the reversible denaturation process of myoglobin
which has been reported to occur through storage time (Cheah &
Ledward, 1996).
3.3.3. Volatile basic nitrogen
Total volatile basic nitrogen substances (VBN) concentration in
chicken breast meat was used as an index of meat freshness. The
VBN were signicantly reduced to 48.3% by 600 MPa pressure at
day 3 of storage, and this was even higher at day 7 under the same
pressure (70.4%) (Table 2). On day 3, 300 and 450 MPa pressure
signicantly reduced VBN compared to the control, but there was
no difference between the 2 treatments. However, at day 7 a similar
trend occurred; the 450 MPa VBN value was signicantly less than
the 300 MPa VBN value. Increase of VBN content in meat can be
caused by either bacterial or enzymatic degradation of proteins
(Egan, Kirk, & Sawyer, 1981, pp. 185e185). In this study lower VBN
values were associated with a decrease in bacteria count caused by
the increase in applied pressure (Table 1). Therefore, in this
experiment we hypothesize that less microbial growth in chicken
breast llet modulated by high pressure led to reduced protein
decomposition and consequently decreased VBN values improving
meat freshness.
3.3.4. Textural prole analysis (TPA)
In our experiment, all textural prole analysis parameters
including cohesiveness, gumminess, hardness and chewiness,
increased with increasing pressure. The 450 and 600 MPa pressure
inicted the most detrimental effects (Table 2). Hardness
increased signicantly at 450 MPa and was not different from
600 MPa pressure. A similar trend was observed for chewiness.
Cohesiveness signicantly increased at 300 MPa and was not
different from 450 to 600 MPa, whereas gumminess signicantly
increased at 450 and 600 MPa compared to controls, but was not
different from 300 MPa pressure treatment (Table 2). Our texture
analysis results are in agreement with other studies. Villacis,
Rastogi, and Balasubramaniam (2008) reported that when turkey
breast muscles were treated with pressure above 150 MPa,
product hardness, gumminess, and chewiness values increased
with increasing pressure. Cohesiveness also increased with the
pressure holding time for all pressures. A similar effect of
increased pressure on hardness was observed in sh meat (Master,
Stegeman, Kals, & Bartels, 2000) and beef muscles (Ma & Ledward,
2004) Hardness is the most important texture attribute to
consumers and dictates the commercial value of a meat
Z.A. Kruk et al. / Food Control 22 (2011) 6e12 10
(Chambers & Bowers, 1993). Interestingly, in our study there was
no effect of pressure on sensory texture evaluation (Fig. 1). This
may be due to cooking before the evaluation. However, negative
and moderate correlations between measurements of cohesive-
ness and resilience, and texture as assessed by sensory panel were
detected (r 0.46 and r 0.56, respectively).
3.3.5. Lipid oxidation (TBARS)
Lipid oxidation as measured by TBAR was affected by
pressure after all storage times (day 0, 3, and 7). In general,
higher pressures increase TBAR values. The most detrimental
effect of pressure was observed at 450 and 600 MPa, whereas
there was no signicant difference between the control and the
300-MPa treatment. This nding is in agreement with the
results of Cheah and Ledward (1996) who observed no signif-
icant increased rate of oxidation in minced pork pressurised at
300 MPa. However, the intensity of oxidation increased with
pressure above 300 MPa. Pressure of 450 MPa in our experi-
ment signicantly increased TBAR values which were almost
doubled at 600 MPa (Table 2). The signicant increase of TBAR
at 450 MPa is a close value to 500 MPa that has been stipulated
to be a critical pressure that initiates lipid oxidation of chicken
breast llet (Orlien, Hansen, & Skibsted, 2000). Further increase
of TBAR values with increased pressure is also in agreement
with Wiggers, Kroger-Ohlsen, and Skibsted (2004). In their
experiment high pressure treatment at 400 MPa and especially
600 MPa caused a signicant increase in secondary lipid
oxidation products although it appeared in cook products
whereas our observations were based on raw product. Orlien
et al. (2000) have shown in chicken breast muscle (llet) that
pressure treatments at 600, 700 and 800 MPa for 5 or 10 min
resulted in some lipid oxidation during storage, and the pres-
sure of 800 MPa was the most damaging. High pressure treat-
ment of raw chicken breast llet induces lipid oxidation at
approximately 450 MPa pressure and this effect has been
magnied at 600 MPa pressure that can last for 7 days of
storage.
4. Conclusions
A pressure of 450 MPa inactivated E. coli, L. monocytogenes in
chicken breast llets to undetectable levels, and reduced S. typhi-
murium by >3 log (CFU/g). The 600 MPa pressure inactivated all
microorganisms below delectable levels. Additionally, the 600 MPa
treatment reduced bacterial count by 6e8 log (CFU/g) improving
shelf-life for 7e14 days. The 450 MPa treatment reduced bacteria
count by 4e8 log (CFU/g), extending shelf-life for 3e14 days,
depending on the micro-uora present. The most susceptible
microorganism to pressure was L. monocytogenes, followed by
E. coli and S. typhimurium. The increased pressure also impacted
sensory characteristics of chicken breast llet. Flavour, aroma
strength and juiciness were the major characteristics affected,
although in a non-consistent manner. Moreover, increasing pres-
sure increased moisture content and cooking loss which could be
the main contributors to the weight loss that occurred at 600 MPa
pressure. Higher pressures caused discolouration of meat by
increasing L*, a* and b*, and increased textural characteristics of
meat such as hardness, cohesiveness, gumminess and chewiness.
However, the higher pressures improved the freshness of meat as
measured by VBN substances.
Further research is needed in order to better understand the
textural, structural, physiochemical and morphological changes
occurring in pressurised meat, and the effect of these changes on
sensory characteristics of chicken breast llets.
References
A.O.A.C. (1990). Ofcial methods of analysis (15th revised ed.). Washington, DC:
Association of Ofcial Analytical Chemists.
Ahn, D. U., Olson, D. G., Jo, C., Love, J., & Jin, S. K. (1999). Volatile production and lipid
oxidation in irradiated cooked sausage as related to packaging and storage.
Journal of Food Science, 64, 226e229.
Ananth, V., Dickson, J. S., Olson, D. G., & Murano, E. A. (1998). Shelf life extension,
safety, and quality of fresh pork loin treated with high hydrostatic pressure.
Journal of Food Protection, 61, 1649e1656.
Ashie, I. N. A., & Simpson, B. K. (1996). Application of high hydrostatic pressure to
control enzyme related fresh seafood texture deterioration. Food Research
International, 29(5e6), 569e575.
Bourne, M. C. (1978). Texture prole analysis. Food Technology, 33, 62e66.
Campus, M., Addis, M. F., Cappuccinelli, R., Porcu, M. C., Pretti, L., Tedde, V., et al.
(2010). Stress relaxation behaviour and structural changes of muscle tissues
from Gilthead Sea Bream (Sparus aurata L.) following high pressure treatment.
Journal of Food Engineering, 96, 192e198.
Carlez, A., Veciana-Nogues, T., & Cheftel, J. C. (1995). Changes in colour and
myoglobin of minced beef meat due to high pressure processing. Lebensmittel
Wiss U Technologie, 28, 528e538.
Chambers, E., & Bowers, J. R. (1993). Consumer perception of sensory qualities in
muscle foods. Food Technology, 47, 116e120.
Cheah, P. B., & Ledward, D. A. (1996). High pressure effects on lipid oxidation in
minced pork. Meat Science, 43(2), 123e134.
Cheftel, J. C. (1995). Review: high-pressure microbial inactivation and food pres-
ervation. Food Science and Technology International, 1, 75e90.
Conway, E. J. (1950). Mircodiffusion and volumetric error (3rd ed.). London: Crosby
Lockwood and Son Ltd.
Crehana, C. M., Troya, D. J., & Buckley, D. J. (2000). Effects of salt level and high
hydrostatic pressure processing on frankfurters formulated with 1.5 and 2.5%
salt. Meat Science, 55, 123e130.
Egan, H., Kirk, R. S., & Sawyer, R. (1981). Pearsons chemical analysis of foods (8th ed.).
UK, Essex: Longman scientic and Technical.
Gola, S., Mutti, P., Manganelli, E., Squarcina, N., & Rovere, P. (2000). Behaviour of
E. coli O157:H7 strains in model system and in raw meat by HPP: microbial and
technological aspects. High Pressure Research, 19, 91e97.
Goutefongea, R., Rampon, V., Nicolas, N., &Dumont, J. P. (1995). InAmericanMeat Science
Association. (Ed.), 41st ICoMST. Meat colour changes under high pressure treatment,
Vol. II (pp. 384e385). San Antonio, TX: American Meat Science Association.
Gross, M., & Jaenicke, R. (1994). Proteins under pressure: the inuence of high
hydrostatic pressure on structure, function and assembly of proteins and
protein complexes. European Journal of Biochemistry, 221, 617e630.
Hansen, E., Trinderup, R., Hviid, M., Darr, M., & Skibsted, L. (2003). Thaw drip loss
and protein characterization of drip from air-frozen, cryogen-frozen, and
pressure-shift-frozen pork longissimus dorsi in relation to ice crystal size.
European Food Research and Technology, 218(1), 2e6.
Hayman, M. M., Baxter, I., ORiordan, P. J., & Stewart, C. M. (2004). Effects of high-
pressure processing on the safety, quality, and shelf life of ready-to-eat meats.
Journal of Food Protection, 67(8), 1709e1718.
Huff-Lonergan, E., & Lonergan, S. M. (2005). Mechanisms of water-holding capacity
of meat: the role of postmortem biochemical and structural changes. Meat
Science, 71, 194e204.
Hugas, M., Garriga, M., & Monfort, J. M. (2002). New mild technologies in meat
processing: high pressure as a model technology. Meat Science, 62, 359e371.
Jung, S., Ghoul, M., & de Lamballerie Anton, M. (2000). Changes in lysosomal
enzyme activities and shear values of high pressure treated meat during ageing.
Meat Science, 56(3), 239e246.
Kim, Y. I., Lee, E. J., Lee, N. H., Kim, Y. H., & Yamamoto, K. (2007). Effects of
Hydrostatic Pressure Treatment on the phsiochemical, morphological, and
textural properties of bovine semitendinosus muscle. Food Science and Biotech-
nology, 16(1), 49e54.
Kristensen, L., & Purslow, P. P. (2001). The effect of ageing on the water-holding
capacity of pork: role of cytoskeletal proteins. Meat Science, 58, 17e23.
Lee, E. J., Kim, Y. J., Lee, N. H., Hong, S. I., & Yamamoto, K. (2007). Differences in
properties of myobrillar proteins from bovine semintendinosus muscle after
hydrostatic pressure or heat treatment. Journal of the Science of Food and
Agriculture, 87, 40e46.
Ma, H. J., & Ledward, D. A. (2004). High pressure/thermal treatment effects on the
texture of beef muscle. Meat Science, 68, 347e355.
Malicki, A., Sysak, Z., & Bruzewicz, S. (2005). Pressurization effect on Salmonella sp
within the sh meal. Bulletin Veterinary Institute Pulawy, 49, 215e217.
Marcos, B., Aymerich, T., & Garriga, M. (2005). Evaluation of high pressure processing
as an additional hurdle to control Listeria monocytogenes and Salmonella enterica
in low-acid fermented sausages. Journal of Food Science, 70(7), 339e344.
Mariutti, L. R. B., Orlien, V., Bragagnolo, N., & Skibsted, L. H. (2008). Effect of sage
and garlic on lipid oxidation in high-pressure processed chicken meat. European
Food Research and Technology, 227, 337e344.
Master, A. M., Stegeman, D., Kals, J., & Bartels, P. V. (2000). Effects of high pressure
on colour and texture of sh. High Pressure Research, 19, 109e115.
Melody, J. L., Lonergan, S. M., Rowe, L. J., Huiatt, T. W., Mayes, M. S., & Huff-
Lonergan, E. (2004). Early postmortem biochemical factors inuence tender-
ness and water holding capacity of three porcine muscles. Journal of Animal
Science, 82, 1195e1205.
Z.A. Kruk et al. / Food Control 22 (2011) 6e12 11
Mor-Mur, M., & Yuste, J. (2003). High pressure processing applied to cooked sausage
manufacture: physical propertiesandsensoryanalysis. Meat Science, 65(3),1187e1191.
Ohshima, T., Ushio, H., & Koizumi, C. (1993). High-pressure processing of sh and
sh products. Trends in Food Science and Technology, 4, 370e375.
Orlien, V., Hansen, E., &Skibsted, L. H. (2000). Lipidoxidationinhigh-pressure processed
chicken breast muscle during chill storage: critical working pressure in relation to
oxidation mechanism. European Food Research and Technology, 211, 99e104.
Patterson, M. F., Quinn, N., Simpson, R., & Gilmour, A. (1995). Effects of high pressure
on vegetative pathogens. In D. A. Ledward, D. E. Johnston, R. G. Earnshaw, &
A. P. M. Hasting (Eds.), High pressure processing of foods (pp. 47e63). Notting-
ham University Press.
Ramirez-Suarez, J. C., & Morrissey, M. T. (2006). Effect of high pressure processing
(HPP) on shelf life of albacore tuna (Thunnus alalunga) minced muscle. Inno-
vative Food Science and Emerging Technologies, 7(1e2), 19e27.
Rastogi, N. K., Raghavaro, K. S. M. S., Balasubramaniam, V. M. M., Niranjan, K., &
Knorr, D. (2007). Opportunities and challenges in high pressure processing of
foods. Critical Reviews in Food Science and Nutrition, 47, 69e112.
Rivas-Canedo, A., Fernandez-Garcia, E., & Nunez, M. (2008). Volatile compounds in
fresh meats subjected to high pressure processing: effect of the packaging
material. Meat Science, 81, 321e328.
SAS Institute. (1989) (4th ed.).SAS/STAT Users Guide. Version 6, Vol. 2 Cary, NC: SAS
Institute.
Serra, X., Grbol, N., Gurdia, M. D., Guerrero, L., Gou, P., Masoliver, P., et al. (2007).
High pressure applied to frozen ham at different process stages. Effect on the
sensory attributes and on the colour characteristics of dry-cured ham. Meat
Science, 75, 21e28.
Shigehisa, T., Ohmori, T., Saito, A., Taji, S., & Hayashi, R. (1991). Effects of high
hydrostatic pressure on characteristics of pork slurries and inactivation of
micro-organisms associated with meat and meat products. International Journal
of Food Microbiology, 12, 207e216.
Styles, M. F., Hoover, D. G., & Farkas, D. F. (1991). Response of Listeria monocytogenes
and Vibrio parahaemoliticus to hydrostatic pressure. Journal of Food Science, 56,
1404e1407.
Villacis, M. F., Rastogi, N. K., & Balasubramaniam, W. M. (2008). Effect of high
pressure on moisture and NaCl diffusion into turkey breast. LWT Food Science
and Technology, 41, 836e844.
Weber, G. (1989). Dynamics of oligomeric proteins. Journal of Molecular Liquids, 42,
255e268.
Wiggers, S. B., Kroger-Ohlsen, M. V., & Skibsted, L. H. (2004). Lipid oxidation in high-
pressure processed chicken breast during chill storage and subsequent heat
treatment: effect of working pressure, packaging atmosphere and storage time.
European Food Research and Technology, 219, 167e170.
Zhang, H., & Mittal, G. S. (2008). Effects of high-pressure processing (HPP) on
bacterial spores: and overview. Food Reviews International, 24, 330e351.
Z.A. Kruk et al. / Food Control 22 (2011) 6e12 12