Meat Science 81 (2009) 321328

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Meat Science
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Volatile compounds in fresh meats subjected to high pressure processing: Effect

of the packaging material
Ana Rivas-Caedo, Estrella Fernndez-Garca *, Manuel Nuez
Departamento de Tecnologa de Alimentos, Instituto Nacional de Investigacin y Tecnologa Agraria y Alimentaria (INIA), Carretera de La Corua, km. 7, 28040 Madrid, Spain

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

Article history: The effect of high pressure treatment (400 MPa, 10 min at 12 C) on the volatile prole of minced beef and
Received 2 June 2008 chicken breast, packaged with or without aluminum foil in a multilayer polymeric bag, was investigated.
Received in revised form 30 July 2008 The analysis of the volatile fraction was carried out by dynamic headspace extraction coupled to gas chro-
Accepted 9 August 2008
matography-mass spectrometry. Pressurization produced signicant changes in the levels of some vola-
tile compounds presumably coming from microbial activity. Some alcohols and aldehydes decreased,
while other compounds, such as 2,3-butanedione and 2-butanone, were more abundant in high pressure
Keywords:
processed meats. A signicant migration of compounds from the plastic material was observed, mainly
High pressure processing
Packaging material
branched-chain alkanes and benzene compounds. Two functions built by the principal component anal-
Volatile compounds ysis explained a high percentage of the variance and could be used to separate the samples into four dis-
Beef tinct groups, according to high pressure treatment and packaging material.
Chicken breast 2008 Published by Elsevier Ltd.

1. Introduction
Increasing consumer demand for more secure and shelf-stable
products has prompted the development of new physical treat-
ments. High pressure processing (HPP) is a non-thermal technol-
ogy using pressures up to 1000 MPa for a variable time, to
extend shelf life of food without modifying its sensory properties
(Hendrickx, Ludykhuyze, Van den Broeck, & Weemaes, 1998) and
nutrient content (Cheftel & Culioli, 1997). HPP renders stable
food due to its ability to kill spoilage and pathogenic microor-
ganisms and to inactivate food enzymes (San Martn, Barbosa-
Cnovas, & Swason, 2002; Patterson, 2005). The bactericidal
effectiveness of HPP depends on factors such as process param-
eters, strain and growth stage of microorganisms, and food ma-
trix (Hoover, Metrick, Papineau, Farkas, & Knorr, 1989; Hugas,
Garriga, & Monfort, 2002).
HPP causes changes in meat mechanical properties (Bouton,
Harris, MacFarlane, & O Shea, 1977; Cheftel & Culioli, 1997; Mac-
Farlane, McKenzie, & Turner, 1986; MacFarlane, McKenzie, Turner,
& Jones, 1981). Color of fresh beef varies after HPP (Marcos, Ayme-
rich, & Garriga, 2005; Serra et al., 2007) due to changes in myoglo-
bin, globin denaturation, heme displacement or release, and
ferrous oxidation (Mor-Mur & Yuste, 2003). In contrast to beef
and pork, poultry muscles are not drastically discolored because
of their lower myoglobin content (Hansen, Trinderup, Hviid, Darr,
& Skibsted, 2003). Lipid stability of pressure-treated foods of ani-
* Corresponding author.
E-mail address: nunez@inia.es (E. Fernndez-Garca).
mal origin has been little investigated, and results are contradic-
tory (Beltrn, Pla, Yuste, & Mor-Mur, 2003; Orlien, Hansen, &
Skibsted, 2000; Wiggers, Ohlsen, & Skibsted, 2004).
Foods are HP-treated in a uid that acts as the pressure trans-
mitting medium. The food is previously packaged in polymeric
materials which serve as a barrier. Among commercial plastic
materials, polyethylenes (PE) and polypropylenes (PP) are mostly
used because of their thermosealability and barrier properties to
water (Sajilata, Savitha, Singhal, & Kanetkar, 2007). It is desirable
that the packaging material maintains the overall food avor (Saji-
lata et al., 2007), but plastics may interact with food components
(Hotchkiss, 1997; Risch & Hotchkiss, 1991) and produce undesir-
able effects, such as loss or imbalance of food avor (Gremli,
1996), oxidation, microbial growth, package damage or even safety
problems (Nielsen & Jgerstad, 1994). The extent of these interac-
tions is inuenced by the properties of the polymer, the avor mol-
ecule (molecular weight, polarity) and external conditions
(Nielsen, Jgerstad, & Oste, 1992). The concentration of the sorbed
material (Brody, 2002) and sorbants are also determining factors
(Charara, Williams, Schmidt, & Marshall, 1992).
All these factors make the study of packaging material safety
necessary, as well as of changes in the overall avor (Hugas
et al., 2002). The aim of the present work was to investigate the
changes in the volatile prole of beef and chicken breast when sub-
jected to HPP using a multilayer plastic as packaging material, with
and without intermediate wrapping in aluminum foil. To identify
the compounds coming from the packaging materials, the volatile
fractions of the plastic material and the aluminum foil were also
studied.

0309-1740/$ - see front matter 2008 Published by Elsevier Ltd.

doi:10.1016/j.meatsci.2008.08.008
322 A. Rivas-Caedo et al. / Meat Science 81 (2009) 321328

2. Materials and methods
2.1. Meats, packaging materials and high pressure processing
Two types of minced meat, beef and chicken breast, were pur-
chased at a local supermarket. Food-grade multilayer packaging
material (MLPM) was used for vacuum packaging (BB4L, Cryovac
Sealed Air Corporation, Milano, Italy). The MLPM consists of one
low density polyethylene (LDPE) layer in direct contact with the
food, followed by several ethylene-vinyl acetate (EVA) layers and
one vinylidene chloride (VDC) layer.
Treatments were conducted in triplicate. Each product was di-
vided into 12 equal portions, six of which were directly vacuum
packaged in MLPM bags while the other six were rst wrapped
in aluminum foil (AF) and then vacuum packaged in the MLPM
bags. Per foodstuff, three portions in direct contact with the MLPM
and three portions shielded with AF were pressurized for 10 min at
400 MPa and 12 C in a 3.5 L capacity discontinuous high pressure
apparatus (Model ACIP 6000, ACB, Nantes, France) whereas the rest
of the samples were kept untreated, as controls. After HPP, all sam-
ples were stored at 4 C for three days and then frozen at
35 1 C until analysis.
2.2. Microbiology
A representative fresh meat sample (10 g) was homogenized
with 90 mL of sterile 0.1% (wt/vol) peptone water solution in a Col-
worth Stomacher 400 (A. J. Seward Ltd., London, UK). Decimal dilu-
tions of samples were prepared in the same sterile solution.
Mesophilic aerobic counts and lactic acid bacteria (LAB) were
determined on duplicate plates of plate count agar (Biolife, Milano,
Italy) and M17 agar (Biolife; acidied at pH 5.7 with 1 N HCl),
respectively, incubated for 48 h at 30 C. Gram-negative bacteria
(GNB) were determined on duplicate plates of PMK agar (Biolife)
incubated for 24 h at 30 C and coliforms were determined on
duplicate plates of VRBA agar (Oxoid, Basingstoke, UK) incubated
for 24 h at 37 C.
2.3. Volatile compound analysis
6 C/min to 100 C, 2 min at 100 C, nal ramp at 17 C/min to
230 C and 6 min at 230 C.
Detection was performed by electron impact ionization, with
70 eV ionization energy operating in the full-scan mode at
6.29 scans/s. Source and quadrupole temperatures were 230 and
150 C. Compound identication was carried out by injection of
commercial standards, by spectra comparison using the Wiley7-
Nist05 Library (Wiley & Sons Inc., Germany), and/or by calculation
of linear retention indexes (LRI) relative to a series of alkanes (C5
C19). The sums of abundances of up to four characteristic ions per
compound were used for semi-quantitative determination. The
areas have been referred to the IS (compound peak area multiplied
by 103 and divided by the IS peak area).
Packaging materials, MLPM and AF, were cut into 0.5 cm2 pieces
and 1.3 g samples were subjected to dynamic headspace extraction
and chromatographic analysis using the same procedure as for
meat samples.
2.4. Statistical analysis
Statistics were performed with the SPSS Win 12.0 software
(SPSS Inc., Chicago, IL). Volatile compound areas were subjected
to analysis of variance (ANOVA) using packaging material and high
pressure treatment as main effects. Principal component analysis
(PCA) with Varimax rotation was performed with selected volatile
compounds.
3. Results
3.1. Microbial counts
Counts of microbial groups in beef and chicken subjected to HPP
and storage for 3 d at 4 C are reported in Table 1. Pressurization
caused a dramatic reduction of all microbial groups in both meats.
Low counts remained in HPP meats after storage for 3 d at 4 C in
comparison with the untreated samples. However, the effect of
pressurization was not identical on all bacterial groups. Gram-neg-
ative bacteria and coliforms declined more than mesophilic aerobic
bacteria and lactic acid bacteria, and differences persisted after
storage for 3 d at 4 C.

Before analysis, meat samples were thawed overnight at 5 C 3.2. Volatile fraction
and then cooked. Twenty grams of minced meat were weighed in
a 5 cm internal diameter stainless steel lidded dish and cooked to The optimized extraction method and analytical procedure al-
an inner temperature of 60 C (100 C, 10 min) in an oven (model lowed the identication of 62 compounds in the volatile fraction
201, J.P. Selecta, Barcelona, Spain). of beef and 53 compounds in chicken breast. Table 2 lists the com-
Volatile compounds were analyzed in an automatic dynamic pounds ordered by chemical family together with their chromato-
headspace apparatus (Purge and Trap, HP 7695, Agilent, Palo
Alto, CA) coupled to a gas chromatograph-mass spectrometer Table 1
(HP-MSD HP 5973, Agilent). Ten grams of cooked meat were Mean microbial countse (SD) in control (C) and high pressure processed (HPP,
400 MPa, 10 min, 12 C) minced samples of beef and chicken breast, before and after
homogenized in a mechanical grinder (IKA Labortechnik, Staufen,
storage for 3 d at 4 C
Germany) with 20 g of anhydrous sodium sulfate (Na2SO4) and
20 lL of an aqueous solution of 630 mg/L cyclohexanone (Sig- Product Microbial C day 1 C day 4 HPP day 1 HPP day 4
group
ma-Aldrich, Alcobendas, Spain) as internal standard (IS). An ali-
quot of the mixture (3.5 g) was subjected to dynamic Minced beef MABf 6.53 0.09a 6.87 0.21a 3.79 0.12b 3.50 0.27b
GNBg 5.31 0.01b 6.03 0.16b 1.73 0.17b <1b
headspace for 20 min at 45 C using helium (45 mL/min) with
LABh 6.59 0.05a 6.91 0.26a 3.53 0.08b 3.18 0.56b
10 min of previous equilibration. Volatile compounds were con- 3.94 0.15b 4.52 0.07a <1c <1c
centrated in a Vocarb 4000 trap (Tekmar, Manson, OH) main- Minced MABf 5.93 0.15b 7.55 0.28a 1.45 0.31d 2.94 0.53c
tained at 35 C, with 4 min dry purge, and desorbed during chicken GNBg 6.11 0.10a 6.21 0.04a <1b <1b
2 min at 260 C through a transfer line heated at 200 C, directly breast LABh 5.62 0.03b 6.76 0.04a <1d 2.56 0.86c
2.32 0.45b 3.86 0.06a <1c <1c
into the injection port at 220 C with a split ratio of 20:1 and
a-d
1.4 mL/min helium ow. Chromatography was carried out in a Means followed by different letters differ signicantly (P<0.01).
e
capillary column (60 m long; 0.25 mm i.d.; 0.5 lm lm thick- expressed in log CFU/g (duplicate determinations in triplicate experiments).
Detection level was 1 log CFU/g.
ness; Innowax, Agilent Technologies), with 1 mL/min helium f
MAB, Mesophilic aerobic bacteria.
ow, with the following temperature program: 8 min at 45 C, g
GNB, Gram negative bacteria.
rst ramp 3 C/min to 60 C, 4 min at 60 C, second ramp at h
LAB, Lactic acid bacteria.
 A. Rivas-Caedo et al. / Meat Science 81 (2009) 321328 323

Table 2
Volatile compounds identied in control and high pressure processed (HPP) minced beef (B) and chicken breast (C), and in the plastic packaging material (PM)
Volatile compound LRIa QIb ID c
Origin d
Signicance of effects e

HPP PM HPP  PM
Hydrocarbons
Pentane 500 42, 41, 57, 72 ST, MS B, C, PM ns ns ns
2-Methylpentane 556 43, 42, 71, 41 MS B, C, PM ns ns ns
3-Methylpentane 583 57, 56, 41, 43 MS B, C, PM ns ns ns
Hexane 600 57, 41, 56, 42 ST, MS B, C, PM ns ns ns
Branched-chain alkane 661 56, 57, 41, 43 MS B, C, PM ns ns ns
Methylcyclopentane 678 56, 69, 42, 41 MS B, C, PM ns ns ns
3-Methylheptane 764 43, 57, 85, 56 MS B, PM ns * ns
Heptane 700 43, 57, 71, 41 ST, MS B, PM ns ns ns
Octane 800 85, 43, 71, 56 ST, MS B, C, PM ns ns ns
2,2,4,6,6-Pentamethylheptane 950 57, 56, 71, 85 ST, MS B, C, PM ns ***/** ns
Branched-chain alkane 1016 57, 71, 43, 70 MS B, PM ns *** ns
Branched-chain alkane 1043 57, 71, 70, 56 MS B, PM ns *** ns
Undecane 1100 57, 71, 41, 56 ST, MS B, PM ns *** ns
Tridecane 1300 57, 71, 43 ST, MS B, PM ns ** ns
Aldehydes
Ethanal 708 44, 43, 42, 41 MS B, C, PM ns/*** ns ns
Propanal 803 58, 57 MS B * * ns
2-Methylpropanal 819 41, 43, 72 ST, MS C *** ns ns
Pentanal 991 58, 44, 41, 57 MS B, PM ns ns ns
Hexanal 1104 56, 57, 72 ST, MS B, C, PM ns ns ns
Octanal 1310 57, 56, 55, 84 MS B, C */ns ns ns
Nonanal 1416 57, 98, 70, 82 MS B, C **/ns */ns ns
Decanal 1525 57, 41, 55 MS B, C ns ns ns
Ketones
2-Propanone 823 58, 43, 42, 39 MS B, C ns/*** ns ns
2-Butanone 911 43, 72, 57 ST, MS B, C */*** ns/*** ns/**
2,3-Butanedione 989 43, 86, 42, 87 MS B, C *** */*** ns/**
3-Hydroxy-2-butanone 1318 45, 43, 88, 42 MS B *** ns ns
2-Cyclohexen-1-one 1487 68, 96, 39, 40 MS B, C ***/ns **/ns ns
Alcohols
2-Propanol 933 43, 41, 45, 59 MS B, C */*** ns/*** ns/*
Ethanol 943 45, 46, 43, 41 MS B, C *** * ns/**
2-Methyl-1-propanol 1120 41, 43, 42, 74 ST, MS C ns * **
1-Propanol 1057 59, 42, 60, 41 MS C *** ** ns
1-Methoxy-2-propanol 1159 45, 47, 43, 75 MS B, C ns ns/*** ns
1-Butanol 1169 56, 41, 43, 42 MS B, C ns **/*** ns/*
3-Methyl-1-butanol 1234 55, 70, 42, 43 ST, MS B, C ns/*** ** *
1-Pentanol 1272 42, 55, 70, 57 MS B, C */ns */** ns/*
3-Methyl-3-buten-1-ol 1274 56, 68, 67, 41 MS B, C * ns/* ns
3-Methyl-2-buten-1-ol 1338 71, 39, 41, 43 MS B ns ns ns
1-Hexanol 1366 56, 55, 69, 41 MS B, C * ns/* ns
1-Heptanol 1464 70, 56, 55, 69 MS B, C ns ns ns
2-Butoxyethanol 1421 57, 87, 41 MS B, C ns ns ns
1-Octen-3-ol 1458 57, 72, 55 ST, MS B, C ns ns ns
2-Ethylhexanol 1496 57, 43, 55, 70 MS B, C ns/* ns/*** ns
1-Octanol 1567 56, 55, 69, 70 MS B, C ns **/ns * / ns

Esters
Ethyl acetate 899 43, 45, 61, 70 MS B, C ns/*** ns/* ns/*
Ethyl lactate 1364 45, 75, 103, 47 MS C *** ns *
Benzyl acetate 1746 108, 91, 150, 90 MS B, C ns ns ns
Benzene compounds
Toluene 1064 91, 92 MS B, C, PM **/ns **/ns ns
Benzene compound 1148 91, 106 MS B, PM ** ns ns
m-Xylene 1163 91, 106, 105, 77 ST, MS B, PM ** ns ns
o-Xylene 1214 91, 106 ST, MS B, PM *** ns ns
Trimethylbenzene 1257 105,120 MS B, PM *** *** ns
Styrene 1287 104, 103 ST, MS C ns ns *
Benzene compound 1308 105, 120 MS B, PM *** ** ns
1,3-Bis(1,1-dimethylethyl)benzene 1450 175, 190, 176, 147 MS B, C, PM ns *** ns
Dichlorobenzene 1485 146, 148, 111 MS B, PM ** ns ns
Benzaldehyde 1578 77, 105, 106, 51 ST, MS B, C ns ns ns
Acetophenone 1690 105, 77, 120, 51 ST, MS B, C ns ns/** ns
Miscellaneous
Carbon disude 734 76, 78, 77, 64 MS B, C ns */ns ns
Dimethyl sulde 753 62, 47, 46, 61 MS B, C ns/*** ns ns
Tetrahydrofuran 870 42, 72, 41, 71 MS B, C ns ns ns
Acetonitrile 1022 41, 40, 43, 39 MS B, C ns ns ns
Chloroform 1039 83, 85, 47, 87 MS B, C ns ns ns
Limonene 1223 68, 93, 67, 121 MS B, PM ns ns ns
4-Terpineol 1643 71, 111, 93, 154 MS C ns ns ns
Menthol 1671 81, 95, 71, 123 MS B, C */ns ns ns
(continued on next page)
324 A. Rivas-Caedo et al. / Meat Science 81 (2009) 321328

Table 2 (continued)
Volatile compound LRIa QIb ID c
Origin d
Signicance of effects e

HPP PM HPP  PM
a-Terpineol 1726 93, 121, 136, 81 ST, MS B, C ns ns ns
Naphthalene 1802 128, 127, 129 MS B, C */ns ns ns
2-Methylnaphthalene 1914 142, 141, 115, 143 MS B, C */ns */ns ns
1-Methylnaphthalene 1862 142, 141,115 MS C ns ns ns
Benzothiazole 1998 135, 108, 69 ST, MS C ns ns ns
a
LRI: Linear retention indexes, calculated in relation to the retention time of n-alkane (C5C19) series.
b
QI, Ions used for quantication.
c
Peak identication: ST, comparison of spectra and retention time with commercial standards; MS, tentatively identied by spectra comparison using the Wiley library.
d
Matrix origin: B, beef; C, chicken breast; PM, multilayer packaging material.
e
ns, non signicant; *, P < 0.05; **, P < 0.01; ***, P < 0.001. Different signicant effects for beef and chicken breast are presented in the order beef/chicken.

graphic indexes, the ions used for quantitation and the signicance
of the effects. A column has been added showing the material in
which compounds were found, beef, chicken breast or plastic
material. Because of the similarity of their fragmentation patterns,
the identication of the branched-chain alkanes, mostly coming
from plastic material, must be taken with caution. As shown in Ta-
ble 2, the most abundant families in both meats were aldehydes
and alcohols. Among all the identied compounds, only a few were
affected by treatments. Results are presented considering sepa-
rately the statistical signicance of the effects.
3.3. Effect of high pressure processing
HPP had a signicant effect on the abundances of 22 volatile com-
pounds in beef (Table 3) and 9 volatile compounds in chicken breast
(Table 4). In beef, the abundance of some aldehydes (3 out of 7, prop-
anal, octanal and nonanal), alcohols (5 out of 16, ethanol, 1-pentanol,
1-hexanol, 2-propanol and 3-methyl-3-buten-1-ol), and benzene
compounds (7 out of 11, toluene, two identied benzene compounds
LRI 1148 and 1308, m-xylene, o-xylene, trimethylbenzene and
dichlorobenzene), was signicantly lower in HP-treated samples
than in control samples (Table 3). Conversely, most ketones (2-buta-
none, 2,3-butanedione and 3-hydroxy-2-butanone), showed signif-
icantly higher levels in HP-treated samples. The behavior of 3-
methyl-1-butanol after HPP depended on the packaging material
(signicant interaction HPP  packaging material), since HP-treated
beef samples showed lower levels of this compound than untreated
samples when previously wrapped in AF, and higher levels than un-
treated samples when directly packaged in MLPM (Table 3).
In chicken breast, the abundance of some aldehydes (ethanal,
octanal, 2-methylpropanal), alcohols (ethanol, 1-propanol, 1-hexa-

Table 4
Table 3 Mean abundancesa (SD) of the volatile compounds in chicken breast signicantly
Mean abundancesa (SD) of the volatile compounds in beef signicantly affected by affected by high pressure processing (HPP)
high pressure processing (HPP)
Volatile compound Packaging material Control HPP
Compounds Packaging Control HPP
material Aldehydes
Ethanal 161.4 30.95 78.20 18.28
Aldehydes Octanal 0.16 0.11 0.00 0.00
Propanal 3.56 0.91 2.51 1.32 2-Methylpropanal 1.05 0.29 0.21 0.21
Octanal 0.73 0.41 0.19 0.31
Nonanal 4.04 1.38 2.59 0.65 Ketones
2-Propanone 46.16 12.21 123.9 14.48
Ketones 2-Butanone MLPMb 7.52 0.67 18.32 0.54
2-Butanone 22.85 3.56 35.90 10.07 AFc + MLPM 6.38 0.15 11.69 1.63
2,3-Butanedione 65.66 58.90 315.1 1029 2,3-Butanedione MLPM 33.01 5.67 39.31 8.90
3-Hydroxy-2-butanone 5.43 7.39 39.10 14.41 AF + MLPM 50.20 13.03 102.4 5.18
2-Cyclohexen-1-one 8.00 0.70 5.45 0.56 Alcohols
Alcohols Ethanol MLPM 25044 1027 6008 1358
Ethanol 4600 941.9 102.0 26.27 AF + MLPM 23414 2369 12142 909.5
1-Pentanol 31.69 19.85 17.37 7.43 1-Propanol 14.74 3.84 3.40 1.49
1-Hexanol 9.30 9.09 2.79 0.89 1-Butanol MLPM 10.63 0.22 9.38 0.59
2-Propanol 35.01 3.91 28.17 4.32 AF + MLPM 7.94 0.64 8.34 0.39
3-Methyl-3-buten-1-ol 3.32 0.54 2.68 0.29 1-Pentanol MLPM 11.38 0.63 9.88 1.23
3-Methyl-1-butanol MLPM b 0.70 0.18 1.04 0.54 AF + MLPM 13.03 0.75 14.94 1.53
AFc + MLPM 2.10 0.07 1.22 0.25 1-Hexanol 4.92 0.93 3.58 0.96
2-Propanol MLPM 256.3 13.86 363.2 50.71
Benzene compounds
AF + MLPM 314.7 27.95 544.4 36.22
Toluene 6.98 1.48 5.25 1.60
2-Ethylhexanol 171.2 38.37 194.0 46.60
Benzene compound LRI 1148 2.08 0.83 0.92 0.48
2-Methyl-1-propanol MLPM 11.41 1.64 6.49 0.90
m-Xylene 6.44 2.35 3.17 1.15
AF + MLPM 10.69 2.58 12.68 0.54
o-Xylene 0.96 0.24 0.25 0.11
3-Methyl-3-buten-1-ol 1.07 0.12 1.23 0.17
Trimethylbenzene 0.73 0.26 0.32 0.27
Ethyl esters
Benzene compound LRI 1308 1.76 0.52 0.99 0.37
Ethyl acetate MLPM 134.5 15.06 2.60 0.86
Dichlorobenzene 0.45 0.07 0.88 0.40
AF + MLPM 92.41 20.06 8.20 1.23
Miscellaneous Ethyl lactate MLPM 24.35 2.44 1.63 0.30
Naphthalene 5.22 1.17 4.01 0.37 AF + MLPM 21.32 2.06 3.66 0.82
2-Methylnaphthalene 2.05 0.45 1.63 0.24 Sulphur compounds
Menthol 1.30 0.12 1.58 0.25 Dimethyl sulde 3.83 0.89 8.46 0.87
a a
Means of triplicate experiments. Separate mean values are given when the Means of triplicate experiments. Seperate mean values are given when the
interaction HPP  packaging material was statistically signicant. interaction HPP  packaging material was statistically signicant.
b b
MLPM, multilayer packaging material. MLPM, multilayer packaging material.
c c
AF, aluminum foil. AF, aluminum foil.
 A. Rivas-Caedo et al. / Meat Science 81 (2009) 321328 325

Table 5 Table 6
Mean abundancesa (SD) of the volatile compounds in beef meat signicantly affected Mean abundancesa (SD) of the volatile compounds in chicken breast signicantly
by the packaging material affected by the packaging material

Volatile compound HPPb/ MLPMc AFd + MLPM Volatile compound MLPMb AFc + MLPM
control
Hydrocarbons
Hydrocarbons 2,2,4,6,6-Pentamethylheptane 20.95 9.05 0.48 0.19
Undecane 2.79 0.74 0.41 0.12
Alcohols
Tridecane 2.02 0.77 1.10 0.31
1-Propanol 11.16 7.24 6.97 5.58
3-Methylheptane 1.43 0.81 3.07 1.11
1-Hexanol 3.73 1.30 4.77 0.72
2,2,4,6,6-Pentamethylheptane 298.5 75.87 8.18 7.99
2-Ethylhexanol 219.4 16.77 145.8 20.65
Branched alkane LRI 1016 23.23 6.30 0.11 0.28
3-Methyl-3-buten-1-ol 1.06 0.09 1.24 0.18
Branched alkane LRI 1043 14.56 4.53 0.05 0.12
1-Methoxy-2-propanol 54.26 11.02 24.71 2.72
Aldehydes
Propanal 3.61 1.04 2.28 1.09 Benzene compounds
Nonanal 3.75 1.49 2.68 0.63 1,3-Bis(1,1-dimethylethyl)benzene 4.70 1.85 0.03 0.06
Ketones Acetophenone 1.56 0.17 1.16 0.15
2,3-Butanedione 255.1 161.8 135.7 125.5 a
Means of triplicate determinations.
2-Cyclohexen-1-one 6.10 1.28 7.25 1.49 b
MLPM, multilayer packaging material.
Alcohols c
AF, aluminum foil.
Ethanole Control 520.9 101.4 3991.4 281.27
HPP 120.83 10.84 76.88 15.41
1-Butanol 4.57 1.17 2.95 0.48 while 1-hexanol and 3-methyl-3-buten-1-ol were found at signi-
1-Pentanol 30.90 18.91 15.91 4.28
1-Octanolf Control 3.89 0.44 2.39 0.08
cantly lower levels in the same samples.
HPP 2.71 0.45 2.52 0.43
3-Methyl-3-buten-1-ol 0.90 0.14 1.09 0.11 3.5. Volatile compounds identied in packaging materials
Benzene compounds
Toluene 7.07 1.54 4.85 1.07
A high number of branched-chain hydrocarbons and benzene
Trimethylbenzene 0.70 0.28 0.29 0.25
Benzene compound LRI 1308 1.63 0.61 1.01 0.36 compounds was observed in the volatile fraction of MLPM (Table
1,3-Bis(1,1- 36.53 7.24 12.65 2.00 2), the most abundant being 2,2,4,6,6-pentamethylheptane and
dimethylethyl)benzene 1,3-bis(1,1-dimethylethyl)benzene. Linear hydrocarbons (C5C16),
Miscellaneous unsaturated hydrocarbons and some linear aldehydes (ethanal,
2-Methylnaphthalene 1.99 0.46 1.63 0.20
Carbon disulde 15.09 2.84 10.36 2.74
pentanal and hexanal) were also found. Only traces of 10 com-
pounds were found in the AF (data not shown), most of which
a
Means of triplicate experiments. are ubiquitous compounds commonly found in laboratories, such
b
HPP, high pressure processing.
c as acetonitrile and tetrahydrofuran.
MLPM, multilayer packaging material.
d
AF, aluminum foil.
e
Signicant effect of packaging material in the 2-way ANOVA. Separate means 3.6. Principal component analysis (PCA)
are shown.
f
Interaction HPP  packaging material statistically signicant. Separate means
Figs. 1 and 2 represent the principal component analysis of
are shown.
some signicant volatile compounds found, respectively, in beef
and chicken breast. The loading plots of the factor scores extracted
by the PCA are represented in Fig. 3 (beef samples) and Fig. 4
nol), and ethyl esters, was signicantly lower after pressurization (chicken breast samples).
(Table 4), and that of most ketones (2-propanone, 2-butanone, In beef (Fig. 1), levels of ethanol and xylene (positive coef-
2,3-butanedione) and other alcohols (2-propanol, 2-ethylhexanol cient), and diacetyl, 3-hydroxy-2-butanone (acetoin) and 2-buta-
and 3-methyl-3-buten-1-ol) signicantly higher in HP-treated none (negative coefcient) build function 1, explaining 50.99% of
samples. It is worth highlighting that the notable decrease caused the variance. The branched-chain hydrocarbons 2,2,4,6,6-pentam-
by HPP in ethanol and derived ethyl esters was more pronounced
in chicken breast samples when directly packaged in MLPM. On
Branched- Pentamethylheptane
the other hand, 1-butanol, 1-pentanol and 2-methylpropanol were 1.0
BDEBa
chain alkanes
more abundant in HP-treated chicken breast samples than un-
treated samples when protected with AF (Table 4).
PC 2 (37.74 % variance)

0.5
3.4. Effect of packaging material
Diacetyl m-Xylene

Acetoin o-Xylene
Higher levels of hydrocarbons and other compounds were ob-
Ethanol
served in beef when directly packed in MLPM (Table 5), namely 0.0
undecane, tridecane, 2,2,4,6,6-pentamethylheptane, two unidenti- 2-Butanone
ed branched-chain hydrocarbons (LRI 1016 and 1043), propanal,
nonanal, 2,3-butanedione (diacetyl), ethanol, 1-butanol, 1-penta-
-0.5
nol, toluene, trimethylbenzene, one unidentied benzene com-
pound (LRI 1308), 1,3-bis(1,1-dimethylethyl)benzene, 2-
methylnaphthalene and carbon disulde. Lower levels of 3-meth-
ylheptane, 2-cyclohexen-1-one and 3-methyl-3-buten-1-ol were -1.0
observed in samples directly packaged in MLPM.
-1.0 -0.5 0.0 0.5 1.0
The relative abundance of some compounds increased in chick-
PC 1 (50.99 % variance)
en breast directly packaged in MLPM, namely 2,2,4,6,6-pentameth-
ylheptane, 1-propanol, 2-ethylhexanol, 1-methoxy-2-propanol, Fig. 1. Loading plot of beef volatile compounds in the rotated space determined by
1,3-bis(1,1-dimethylethyl)benzene and acetophenone (Table 6), the principal components. a1,3-bis(1,1-dimethylethyl)benzene.
326 A. Rivas-Caedo et al. / Meat Science 81 (2009) 321328

1.0
Pentamethylheptane HPP
BDEBa
PC 2 (29.90 % variance)

1.0 Control
0.5 HPP
Ethyl acetate
Control
2-Butanone

Regression factor score 2

Control
Ethyl lactate
HPP
Ethanol
0.0 Control
2-Methyl-1-propanol 0.0
2-Propanone
Control
3-Methyl-1-butanol
-0.5
Control
3-Methyl-3-butenol HPP
-1.0

Diacetyl HPP
-1.0

-1.0 -0.5 0.0 0.5 1.0 HPP

PC 1 (60.42 % variance)
Fig. 2. Loading plot of chicken breast volatile compounds in the rotated space
determined by the principal components. a1,3-bis(1,1-dimethylethyl)benzene.
ethylheptane and those with LRI 1016 and 1043, together with the
benzene compound 1,3-bis(1,1-dimethylethyl)benzene make up
function 2, explaining 37.74% of the variance.
In chicken breast (Fig. 2), the levels of ethanol, but also those of
derived ethyl esters, and branched-chain molecules 2-methylprop-
anal and 3-methyl-1-butanol (with positive coefcient), and 2-
propanone and 2-butanone (with negative coefcient), compose
function 1, explaining 60.42% of the variance. 2,2,4,6,6-Pentameth-
ylheptane and the benzene compound 1,3-bis(1,1-dimethyl-
ethyl)benzene make up function 2, together with 3-methyl-3-
buten-1-ol, explaining 29.90% of the variance.
In both meats PC1 is related to the HP treatment and PC2 di-
vides the samples according to the packaging material.
4. Discussion
4.1. Effect of HPP on the volatile fraction
Gram-negative bacteria in beef and chicken breast were more
affected by HPP than Gram-positive bacteria, in agreement with
Control
HPP
HPP
1.0
Regression factor score 2
-2.0
-1.5 -1.0 -0.5 0.0 0.5 1.0 1.5
Regression factor score 1
Fig. 4. Loading plot of the factor scores of the chicken breast samples in the plane
dened by principal components. d samples directly packaged in MLPM s, samples
shielded with AF. Values in the gure correspond to HP treatment.
previous reports (Hugas et al., 2002; Smelt, 1998; Garriga, Grbol,
Aymerich, Monfort, & Hugas, 2004). Also, the effect of HPP on vol-
atile compounds differed. It clearly decreased the levels of a num-
ber of compounds coming from microbial metabolism, such as
ethanol and ethyl esters, or from lipid oxidation, such as aldehydes
and 1-alcohols, while favoring the formation of diacetyl and re-
lated compounds such as acetoin and 2-butanone.
As shown in Figs. 14, the volatile compounds and the individ-
ual meat samples are perfectly distributed into well-dened
groups by the Principal Component Analysis. PC1 could be desig-
nated as HP treatment, the untreated samples occupying the right
part of the plane (Figs. 3 and 4) due to their higher contents of eth-
anol and other compounds such as xylene in beef and esters in
chicken, and their lower contents of some ketones, which have
been proved to increase with HPP. PC2 could be named packaging
material. Samples directly packaged in MLPM are pushed to the
upper side of the plane, mainly due to their higher contents of
branched-chain hydrocarbons and benzene compounds. In chick-
Control en, PC2 also separates the HPP samples previously wrapped in AF
(with higher diacetyl levels) from the untreated samples directly
packaged in MLPM (Fig. 4).
Diacetyl is produced during lactose and citrate metabolism (Co-

Control
HPP gan, 1995) by the action of different bacteria, among which the
most important are LAB, but it can also be produced by others such
as staphylococci (Berdagu, Monteil, Montel, & Talon, 1993; Ole-
HPP
0.0 sen, Meyer, & Stahnke, 2004). HPP might have favored the release
of citrate, as well as glucose and its metabolites, from the muscle
cells thus enhancing their availability, a fact which might explain
the observed increase in diacetyl. Also, the higher resistance of
HPP Control LAB to HPP (Table 1), could have positively discriminated diace-
HPP Control
tyl-producing strains. Higher levels of diacetyl were found in HP-
-1.0
treated Hispnico cheese than in untreated cheese (vila, Garde,
HPP Control Fernndez-Garca, Medina, & Nuez, 2006).
Bacteria such as pseudomonads and staphylococci as well as
yeasts are among the main ethyl ester producers. The decrease in
-1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 the levels of these compounds can be related to the killing of bac-
Regression factor score 1 teria (and possibly yeasts) by HPP (Table 1).
Based on studies which have associated HPP with lower lipid
Fig. 3. Loading plot of the factor scores of the beef samples in the plane dened by
principal components. d samples directly packaged in MLPM s, samples shielded
stability in meat (Cheftel & Culioli, 1997; Orlien et al., 2000), an
with AF. Values in the gure correspond to HP treatment. increase of compounds supposedly coming from lipid oxidation,
 A. Rivas-Caedo et al. / Meat Science 81 (2009) 321328
such as aldehydes and alcohols, would have been expected in
HP-treated meats. However, our results clearly show that most
of these compounds were lower in HP-treated samples. Lipid
stability of foods of animal origin subjected to HPP seems to
be dependent on treatment intensity. Some authors (Orlien
et al., 2000) reported higher levels of lipooxidation products in
chicken breast after pressurizing at 800 MPa for 10 min, but no
lipid oxidation was observed when applying less than 500 MPa.
Beltrn et al. (2003) conrmed that lipid stability of pressurized
poultry was related to cell membrane integrity and that pressure
treatments up to 500 MPa did not induce lipid oxidation. Wig-
gers et al. (2004) obtained similar results in chicken breast when
subjected to pressures of 400600 MPa for 5 min, while stressing
the inuence of oxygen availability on the formation of these
products.
The lower levels of compounds like linear aldehydes and 1-alco-
hols found after HP treatment could indicate that, either HPP is
reducing lipid oxidation phenomena in the treated meats, or that
the decrease of these compounds is a direct consequence of the
killing of certain bacteria. Some Gram-negative bacteria, such as
Pseudomonas, Proteus or Aeromonas, are known to have a high lipo-
lytic activity and, as already discussed, are very sensitive to HPP
(Hoover et al., 1989; Rubio, Martnez, Garca-Cachn, Rovira, &
Jaime, 2007; Smelt, 1998).
The consistent decrease of benzene compounds and polyaro-
matic compounds in HP-treated meats deserves comment. The
origin of these compounds is controversial. Some authors have
suggested a feed origin (Buscailhon, Berdagu, & Monin, 1993)
or a pollution origin (Bosset, Gubler, Btikofer, & Gauch,
2000), while others (Morales, Fernndez-Garca, & Nuez,
2005) observed an increase of styrene and dichlorobenzene in
cheeses made from milk inoculated with Pseudomonas fragi,
which might indicate a microbial origin for some benzene com-
pounds. Our results would be consistent with the microbial ori-
gin hypothesis.
4.2. Effect of packaging material
Migration was the main phenomenon occurring in beef and
chicken packaged in MLPM, although the effect was dependent
on the food matrix, being more intense in beef than in chicken
breast. The most abundant compounds identied in the volatile
fraction of MLPM, 2,2,4,6,6-pentamethylheptane and 1,3-bis(1,1-
dimethylethyl) benzene, main factors in the PCA, were observed
at higher levels in meats than were the other hydrocarbons and
benzenic compounds. Trace amounts of those two compounds
were even found in samples previously wrapped in AF. According
to Franz (2000), migration increases with increasing fat content be-
cause most of the plastic material constituents are lipophilic rather
than hydrophilic. The higher fat content in beef in comparison with
chicken breast can explain the more intense migration observed in
beef.
Higher levels of 2-methylnaphthalene were found in beef sam-
ples when directly packaged in MLPM. Compounds such as naph-
thalene are considered to be environmental pollutants. Lau,
Wong, and Leung (1994) observed the absorption of this com-
pound from air to low density polyethylene packaging and after-
wards to the milk which it contained. The increase in the levels
of this compound in beef when no AF was used might indicate that
AF acts as a barrier against the migration of a number of com-
pounds coming from the environment.
It is worth noting that, unexpectedly, the migration of com-
pounds from the MLPM did not seem to be enhanced by HPP.
There is little information on the effect of HPP on the properties
of polymeric packaging materials and possible package-food-
stuff-environment interactions. Studies in which ethylene vinyl
327
alcohol and polyvinyl alcohol materials were HP-treated re-
vealed no signicant changes in their structure (Ochiai & Nak-
agawa, 1992), or even slightly better barrier properties were
observed (Lpez-Rubio et al., 2005), which may be due to a
glassy transition of the polymer (Kbel, Ludwig, Marx, & Tau-
cher, 1996). More research is needed to ascertain the possible
interactions between packaging material and HPP in real food
matrices.
The volatile proles of minced beef and chicken breast sub-
jected to high pressure processing underwent signicant changes,
most of which can be associated with the death of microorganisms
or the inactivation of enzymes related with the formation of avor
compounds. The packaging material is an important factor in
maintaining the volatile prole of treated meats and the use of alu-
minum shielding was considered to be appropriate, since lower
interactions between package, meat and the environment were ob-
served in both HP-treated and untreated meat samples when
wrapped in aluminum foil prior to packaging in a polymeric plastic
material.
Acknowledgments
This work was supported by projects CPE 03-012 (INIA), TEMI-
NYSA S-0505/AGR/0314 (Comunidad de Madrid) and CARNISE-
NUSA CSD 2007-00016 (Consolider). The authors thank INIA for
granting Ana Rivas-Caedo, and C. Juez Ojeda, J. Calzada, B. Rodr-
guez and M. de Paz for their valuable technical assistance.
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