Meat Science 92 (2012) 604–609

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Meat Science
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Influence of vacuum skin packaging on color stability of beef longissimus lumborum

compared with vacuum and high-oxygen modified atmosphere packaging
Xin Li ⁎, Gunilla Lindahl, Galia Zamaratskaia, Kerstin Lundström
Swedish University of Agricultural Sciences, Uppsala BioCenter, Department of Food Science, P.O. Box 7051, SE-750 07, Uppsala, Sweden

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

Article history: The objective of this study was to investigate how color stability of beef is affected by vacuum skin packaging
Received 30 December 2011 (VSP) compared with vacuum packaging (VP) and high-oxygen modified atmosphere packaging (MAP; 80%
Received in revised form 16 April 2012 O2 and 20% CO2). Longissimus lumborum muscles were aged in vacuum for 7 days and then cut into 2-cm-thick
Accepted 7 June 2012
slices and repacked using VSP, VP and MAP for another 7 days. Color stability was measured during the
next 5 days in air and samples for α-tocopherol and NADH analyses were obtained at the beginning and end
Keywords:
Beef
of aerobic storage. Color stability, α-tocopherol and NADH of steaks were affected by packaging methods and
Packaging method storage time in air (P b 0.05). Higher a* value was obtained in VSP on day 5 compared with VP. Steaks packed
Color stability in VSP had better color stability than in VP and their color was similar to MAP at the end (day 5) of storage.
α-Tocopherol © 2012 Elsevier Ltd. All rights reserved.
NADH

1. Introduction color due to deoxymyoglobin (DeoxyMb) formation (Jeremiah, 2001).

One of the most important meat quality aspects determining
consumers' purchase choice is color. Meat discoloration is used by
consumers as an indicator of freshness and wholesomeness (Mancini
& Hunt, 2005). Thus, improvement of color stability is important in
the meat industry.
Most fresh meat is packaged to extend meat shelf life. Various
packaging methods offer different environments for meat products
which may affect meat quality (Ferioli, Caboni, & Dutta, 2008;
Vázquez et al., 2004; Zakrys, Sullivan, Allen, & Kerry, 2009). In Sweden,
modified atmosphere packaging (MAP) with the gas composition
of 80% O2 and 20% CO2 is a common packaging method for beef.
The high oxygen content in MAP gives the beef a stable bright-red
color due to the formation of a thick layer of oxymyoglobin (OxyMb)
at the meat surface that masks the underlaying metmyoglobin
(MetMb) layer (Jeremiah, 2001). In addition oxygen partial pressures
above that which are present in air retard MetMb formation and pre-
serve OxyMb and redness (Jeremiah, 2001). However, some studies
have found that MAP negatively affected the cooked color of meat
(Lagerstedt, Ahnström, & Lundström, 2011; McMillin, 2008) and ten-
derness development (Lagerstedt, Ahnström, et al., 2011; Lagerstedt,
Lundström, & Lindahl, 2011; Lund Nissen, Christensen, Fregil, Hviid,
& Skibsted, 2008). Vacuum packaging (VP) offers anaerobic conditions
for the meat, which extends shelf life. VP provides a stable purple
⁎ Corresponding author. Tel.: + 46 18 671641; fax: + 46 18 672995.
E-mail address: xin.li@slu.se (X. Li).
However, the exudate held in wrinkles in VP may be more susceptible
to bacterial growth and is also not desirable from a consumer perspec-
tive. Vacuum skin packaging (VSP) minimizes the formation of wrinkles
and air pockets by heating the upper cover film and making it shrink
tightly around the meat. Meat in such packages has longer shelf life
and slower bacterial growth compared to VP (Vázquez et al., 2004).
As an antioxidant, α-tocopherol limits the formation of MetMb.
In animal tissues, α-tocopherol has the highest biological activity
compared with other tocopherol forms (Buttriss & Diplock, 1984).
α-Tocopherol competes with OxyMb for lipid peroxyl radicals and
thus inhibits the formation of MetMb. When α-tocopherol is consumed,
MetMb accumulation proceeds without inhibition (Lanari, Schaefer, Liu,
& Cassens, 1996).
Reduced nicotinamide adenine dinucleotide (NADH) plays an im-
portant role in the MetMb reducing process. Metmyoglobin can be
converted to DeoxyMb and/or OxyMb in the presence of NADH enzy-
matically or non-enzymatically (Bekhit & Faustman, 2005; Brown &
Snyder, 1969; Shirabe et al., 1992). Echevarne, Renerre, and Labas
(1990) reported that MetMb reductase activity was not the most
potent factor in the control of meat discoloration rate. Increase of
MetMb reductase activity does not automatically lead to increased
color stability (Seyfert, Hunt, Ahnström, & Johnson, 2007). Bekhit,
Geesink, Ilian, Morton, and Bickerstaffe (2003) concluded that the
lack of NADH was the limiting factor of beef MetMb reducing activity
and, consequently, beef color stability. Although the relationship be-
tween NADH and color stability is well established, information on
the rate of depletion of NADH reservoirs in packed meat is incomplete.
The number of studies on color change after opening the packages and
exposing in air is limited.

0309-1740/$ – see front matter © 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.meatsci.2012.06.006
 X. Li et al. / Meat Science 92 (2012) 604–609
Thus, the aim of this study was to investigate color stability and
contents of α-tocopherol and NADH in beef packaged in VSP com-
pared with VP and MAP. Moreover, the effect of storage in air on
these parameters after opening the packages was investigated. This
study was a part of a larger project on effects of packaging method
on shear force, sensory quality and water holding capacity in beef
(Lagerstedt, Ahnström, et al., 2011).
2. Materials and methods
2.1. Animals and samples
Ten young bulls (16–25 months) of beef breed crosses, from two
farms were slaughtered on the same day using standard routines of
a commercial slaughter plant. The carcasses were electrically stimu-
lated (low voltage, 80 V, pulses of 5 ms duration for 30 s) 30 min
after exsanguination and then kept in a chilling room at 4 °C over-
night. The anterior and posterior carcass quarters were separated
between the 9th and 10th ribs. On day 1 post mortem both sides of the
longissimus lumborum (LL) on each animal were removed from the
10th rib to the last lumbar vertebrae. The pH was measured between
the 9th and 10th ribs with a portable pH meter (Knick Portamess®
913, Berlin, Germany) equipped with a combination pH gel electrode
(SE 104, Knick, Berlin, Germany). All selected LLs had a pH lower than
5.8 at 24 h post mortem.
2.2. Packaging
The LLs were packed in vacuum and aged for 7 days at 2 °C in
darkness in a single layer. The LLs were then unpacked and 2 cm-
thick slices were cut out from the position 30 cm after the 10th rib.
These slices were assigned to the following packaging methods:
(1) vacuum packaging (VP), n = 10. The vacuum bag (Clingvac 90,
Curevac AB, Göteborg, Sweden) was 90 μm thick with transmission
rates (cm 3/m 2, 24 h, 23 °C): O2 40, CO2 150; (2) modified atmosphere
packaging (MAP): 80% O2 + 20% CO2, n = 8. Each sample packed in
MAP was put on plastic straws in the tray to allow oxygen to pene-
trate into the meat evenly from all sides. The lidding film (Cryovac®
LID 2050, Sealed Air Corporation, USA) used in MAP was 25 μm
thick with transmission rates (cm 3/m 2, 24 h, 23 °C): O2 24, CO2 95;
(3) vacuum skin packaging (VSP), n = 5. The top film (Cryovac®
Darfresh® TH300, Sealed Air Corporation, USA) used in VSP was
150 μm thick; its oxygen permeability rate (cm 3/m 2, 24 h, bar at
23 °C, 0% relative humidity) was 2 and moisture vapor transmission
rate (g/m 2, 24 h, 38 °C, 100% relative humidity) was 9. The bottom
film (Cryovac® Darfresh®, Sealed Air Corporation, USA) used in VSP
was 280 μm thick; its oxygen permeability rate (cm 3/m 2, 24 h, bar
at 23 °C, 0% relative humidity) was 2. The samples were stored in
their respective packaging system at 4 °C for an additional 7 days in
darkness in single layers, i.e. in total 14 days post mortem including
the aging time of 7 days of whole LLs in vacuum. The samples were
then exposed to air wrapped with oxygen-permeable PVC-film
(NORM PACK 115 45‐1, Tempac AB, Tyresö, Sweden). The overall mi-
gration rate (mg/dm 2, 24 h, 20 °C) of the PVC-film was 1. In addition,
one slice of LL was kept as a control (n = 10) after the initial aging
time at day 7. The control was exposed to air in darkness at the
time the other treatments started, i.e. in total 7 days post mortem.
2.3. Color
The color of the samples over wrapped with PVC was measured
through the PVC film after blooming at 4 °C for 1.5 h (day 0) and
then on days 1, 3 and 5 of storage in air. Color was measured using
a Minolta CM-2500d spectrophotometer (Konica Minolta Sensing
Inc., Osaka, Japan) with specular reflectance excluded, 8 mm diame-
ter measuring aperture, illuminant D65, 10° standard observer and
605
CIE L*, a*, b* color scale. The measuring aperture was covered with a
glass plate and the instrument was calibrated against a white plate
(L* = 97.29 ± 0.02, a* = − 0.07 ± 0.10, b* = −0.12 ± 0.10). The aver-
age value of four measurements on the meat surface was used. The
Minolta instrument recorded reflectance values in the range of
360 nm to 740 nm at10‐nm intervals. Reflectance values that were
not directly measured by the color instrument at specific wavelengths
(474, 525 and 572 nm) were calculated by linear interpolation. The
Kubelka–Munk K/S values were then calculated. The relative content
of DeoxyMb was estimated by the ratio (K/S474)/(K/S525), the rela-
tive content of OxyMb by the ratio (K/S610)/(K/S525) and the rela-
tive content of MetMb by the ratio (K/S572)/(K/S525) (Hunt et al.,
1991; Mancini, Hunt, & Kropf, 2003). The K/S ratio decreases when
the relative content of the corresponding myoglobin form increases.
Therefore the K/S ratios were transformed in order to get the correct
impression when looking at the figures. The K/S ratio of DeoxyMb was
transformed to [1.5 - (K/S474)/(K/S525)], the K/S ratio of OxyMb was
transformed to [1 - (K/S610)/(K/S525)] and the K/S ratio of MetMb
was transformed to [2 - (K/S572)/(K/S525)].
2.4. α-Tocopherol content
Samples for analysis of α-tocopherol and NADH contents were
collected on day 0 and day 5 of the color measurements. The samples
were cut out from the middle of the 2-cm-thick slices including
both surface and interior. These samples were stored at −80 °C until
analysis.
For α-tocopherol, the method by Högberg, Pickova, Babol, Andersson
and Dutta (2002) was used. Briefly, duplicate samples of 1 g muscle
tissue excluding the surface were chopped into small pieces and
homogenized in 2 ml absolute ethanol using Ultra-Turrax® T 25 Basic
homogenizer (IKA® Werke GmbH & Co. KG, Staufen, Germany). To sa-
ponify the samples, 1.2 ml of 20% (w/v) ascorbic acid, 0.6 ml methanol
and 1.2 ml of 17.9 M KOH were added. Samples were then agitated
twice in a 70 °C water bath for 10 min and vortexed for 10 s during
the interval. After cooling in ice cold water, tocopherol was extracted
twice by 4 ml of hexane. The hexane was evaporated under nitrogen
gas and the tocopherol was dissolved by the mobile phase. The α-
tocopherol content was measured by high performance liquid chroma-
tography (HPLC) with fluorescence detector set at 290 nm excitation
and 327 nm emission wavelength. The mobile phase consisted of 95%
methanol:acetonitrile (1:1 v/v) and 5% chloroform. The α-tocopherol
content was presented as the area under the peak. The α-tocopherol
content in fresh muscle tissue (μg/g) was calculated based from the
standard curve constructed using known amounts of α-tocopherol.
2.5. NADH content
NADH was extracted by alkaline extraction as described by
Klingenberg (1974). Duplicate samples were prepared by combining
1.5 g frozen muscle tissue, excluding the surface with 12 ml of 0.5 M
cooled alcoholic KOH and then vortexing for 30 s. The samples were
agitated in a 90 °C water bath for 5 min to decompose NAD, and
then cooled rapidly to 0 °C in an ice bath for 5 min. To neutralize
each sample, about 11 ml triethanolamine-HCl–phosphate mixture
(0.5 M triethanolamine hydrochloride; 0.4 M KH2PO4; 0.1 M K2HPO4)
was added to adjust the pH to 7.8. After holding at room temperature
for 10 min to flocculate the denatured protein, the sample was
centrifuged at 25,000 ×g for 10 min at 4 °C (Sorvall® Super T 21 Cen-
trifuge, Kendro Laboratory Products, Newtown, USA) to get a clear
supernatant fluid. The NADH concentration was measured using a
spectrophotometric recycling method (McCormick & Wright, 1971).
The assay mixture contained the following: 2.5 ml of 17.5 μg/ml
dichlorophenolindophenol (DCPIP), 0.5 ml of 0.1 M pH 7.4 sodium
phosphate buffer, 0.05 ml of 1 mg/ml phenazine methosulfate (PMS),
0.1 ml ethanol, 0.1 ml muscle extract supernatant and 0.05 ml alcohol
606 X. Li et al. / Meat Science 92 (2012) 604–609

dehydrogenase (E1.1.1.1, Sigma, USA, containing 0.3 mg protein). The
reduction of DCPIP to its colorless leuco base was measured. The
change of absorbance at 600 nm was measured (UV-2401 PC Spectro-
photometer, Shimadzu, Kyoto, Japan) for 20 min to determine the
NADH concentration. The NADH content in fresh muscle tissue
(nmol/g) was calculated from the standard curve using known amounts
of NADH.
2.6. Statistical analysis
Statistical analysis was carried out with the Statistical Analysis
System (Version 9.2, SAS Institute, Cary, NC, USA). The BOXPLOT pro-
cedure was used to describe the data set; the SCHEMATICIDFAR was
used in BOXSTYLE option and the ID option was specified to label
observations outside the lower or upper far fences. The MIXED proce-
dure was used to estimate the effects on variation in meat color
stability of packaging methods, storage time and their interaction
as fixed factors and animal as random factor. To correct for unequal
variances between packaging methods, Satterthwaite's method was
implemented using packaging method as group in the REPEATED
statement. The option PDIFF with Bonferroni adjustment for multiple
comparisons was used to adjust the P-values when multiple compari-
sons were made between different packaging methods within the
same aerobic storage time or between different aerobic storage times
within the same packaging method.
3. Results
The relative content of MetMb was in one of the MAP samples ob-
served to be 1.25 after 5 days of storage in air. This observation was
higher than the upper far fence and labeled an outlier in SAS output.
This may be due to bacterial contamination during aging and storage
leading to fast bacterial growth and spoilage of the sample (bad smell
on day 5 in air). Thus, data from this sample on day 5 were not included
in the statistical analysis.
was higher for samples packed in VSP than in VP on day 5. For samples
packed in VSP, VP and control, higher a* values were observed on day 1
than on days 0 and 5 during exposure in air. The a* values for samples
packed in VSP and VP were higher on day 3 than on day 0 during storage
in air. The b* value was higher for samples packed in MAP than in VP on
day 0. On day 5, the b* value for samples in VSP and MAP was higher
than in the control. A lower b* value was observed in control on day 5
than on the other days in air.
The relative content of DeoxyMb was significantly affected by
packaging methods and interaction of packaging methods and aerobic
storage time (P b 0.05, Fig. 1). On day 0, the relative contents of
DeoxyMb in VP and VSP were higher than in MAP. The relative content
of DeoxyMb in the control was lower than in the other treatments
during storage in air except on day 5. At the end of storage (day 5),
the relative contents of DeoxyMb did not differ among packaging
methods. The relative content of OxyMb was influenced by packaging
method, aerobic storage time and their interaction (P b 0.05, Fig. 1).
The relative contents of OxyMb for samples in VP, VSP and control
were lower than in MAP on day 0. The relative contents of OxyMb
for samples in MAP and VSP were higher than in control samples on
day 5. The relative content of OxyMb for samples packed in VP was
lower than in VSP and tended to be lower than in MAP (P = 0.058)
on day 5. The relative content of OxyMb for samples packed in VP,
VSP and control was higher on day 1 than on day 0. For samples in
VP and control, the relative contents of OxyMb were also higher on
day 1 than on day 5. Packaging method, aerobic storage time and
their interaction affected the relative content of MetMb (P b 0.05;
Fig. 1). The relative contents of MetMb were higher in MAP and con-
trol samples than in VP and VSP samples on day 0, and then during
storage, MetMb contents were stable in MAP and VSP but increased
in control and VP. Thus at the end of storage, the relative contents
of MetMb were lower in MAP and VSP than in control and tended to
be slightly lower in MAP than in VP (P = 0.065).
3.2. α-Tocopherol content

3.1. Color The α-tocopherol content in LL was affected by packaging methods

L*, a* and b* values were significantly affected by packaging method
and aerobic storage time (Table 1). The L* value for samples packed
in MAP was higher than those in the other packaging methods and
the control on day 0. The L* value for samples packed in control
was lower than in those packed in MAP on days 1 and 3. L* values
for samples packed in VP were lower than in MAP on day 3. The L*
value for control samples was higher on day 5 than on days 0 and 1.
The a* value was higher for samples packed in MAP than in the other
packaging methods and those of the control on day 0. The a* value
(P = 0.058), storage time (P b 0.05) and their interaction (P b 0.05,
Table 2). The α-tocopherol content for samples packed in MAP was
lower than in VSP samples on day 0. The α-tocopherol content was
significantly lower on day 5 than on day 0 in all packaging methods
except MAP.
3.3. NADH content
NADH content differed between packaging methods and storage
time (P b 0.05), but there was no interaction between packaging

Table 1
Effect of packaging method (PM) and aerobic storage time (day) on meat color stability.

PM Aging Number Day SE P-value

(day)
0 1 3 5 PM Day PM × Day

L* value (lightness) Control 7 10 34.9bB 35.0bB 35.4abB 37.3a 0.75 b 0.001 b 0.001 0.056
MAP 14 8⁎ 36.7A 36.6A 37.0A 36.7 0.62
VP 14 10 34.5B 35.4AB 35.6B 35.7 0.70
VSP 14 5 35.6B 36.5AB 36.3AB 36.3 0.64
a* value (redness) Control 7 10 16.1bB 18.0a 17.2ab 15.7bABC 0.61 b 0.001 b 0.001 0.006
MAP 14 8⁎ 18.0A 17.8 17.1 17.2AC 0.58
VP 14 10 15.6cB 17.8a 16.8ab 15.9bcC 0.51
VSP 14 5 15.9cB 18.3a 17.5ab 17.1bAB 0.51
b* value (yellowness) Control 7 10 15.9aAB 16.6a 16.4a 14.3bB 0.42 0.001 b 0.001 0.061
MAP 14 8⁎ 17.0A 16.5 16.1 16.0A 0.31
VP 14 10 16.0B 16.1 15.7 15.2AB 0.28
VSP 14 5 16.2AB 16.9 16.5 16.5A 0.40

MAP: modified atmosphere packaging, 80% O2 + 20% CO2; VP: vacuum packaging; VSP: vacuum skin packaging. SE: standard error. Means with different low-case superscripts
within the rows (effect of aerobic storage time) differ at P b 0.05 after Bonferroni correction. Means with different capital superscripts within the columns (effect of PM) differ at
P b 0.05 after Bonferroni correction. Number of comparisons = 6. ⁎One sample on day 5 with outlier value was not included in the results.
 X. Li et al. / Meat Science 92 (2012) 604–609 607

Deoxymyoglobin (1.5 - K/S ratio) Oxymyoglobin (1 - K/S ratio) Metmyoglobin (2 - K/S ratio)
0.63 0.78 0.70
0.65
0.61 0.76
0.60
0.59 0.74 0.55
0.50
0.57 0.72
0.45

0.55 Day 0.70 Day 0.40 Day

0 1 3 5 0 1 3 5 0 1 3 5

Control MAP VP VSP

Fig. 1. Effect of packaging method and dark aerobic storage time (day) on relative pigment contents calculated as K/S ratio. Notice that DeoxyMb is shown as (1.5 - K/S ratio),
OxyMb as (1 - K/S ratio) and MetMb as (2 - K/S ratio). MAP: modified atmosphere packaging, 80% O2 + 20% CO2; VP: vacuum packaging; VSP: vacuum skin packaging. Data points
represent least squares means. Error bars represent positive and negative standard errors. Number of animal in each packaging method: Control (n = 10); MAP (n = 8, data of one
animal on day 5 with outlier value was not included in the result.); VP (n = 10); VSP (n = 5).

methods and storage time (Table 2). The NADH content was signifi-
cantly higher in VSP on day 5 than on day 0. NADH content tended
to be higher in VP on day 5 than on day 0 (P = 0.052). NADH content
in MAP tended to be higher than in VSP (P = 0.051) and in VP
(P = 0.066) on day 0. NADH content did not differ among packaging
methods on day 5.
4. Discussion
In this study, the entire loins were packed in vacuum for 7 days,
then cut into 2-cm-thick slices and repacked using VSP, VP and MAP
for another 7 days. To simulate the change in color when consumers
store beef steaks in the refrigerator after opening the package, the
color of the steaks during 5 days in air was measured. Discoloration
during this period is of interest and may influence consumer satisfac-
tion of the product.
Samples packed in VSP and VP had lower lightness (L* value) than
those packed in MAP when the package was opened after aging for
14 days (day 0 in air), whereas the L* values were similar between
these packaging methods on the last day of exposure in air. The a*
value of LL was higher in MAP than in the other packaging methods
at the beginning of color measurement (day 0 in air). This result
was in agreement with Cayuela, Gil, Bañón, and Garrido (2004) and
may be due to the higher oxygen (80%) content in MAP, which con-
tributed to the oxygenation of myoglobin. The VP and VSP had more
similarities because they both provided a vacuum environment for
the meat, before repacking the samples in air-permeable packaging.
The limited oxygen resulted in higher relative contents of DeoxyMb
in VP and VSP than in control and MAP. The purple color of DeoxyMb
might to a certain extent decrease consumer satisfaction (Carpenter,
Cornforth, & Whittier, 2001). In this study, samples packed in VP,
VSP and control for 7 days were all in vacuum which probably con-
tributed to their lower a* values after 1.5 h of storage in air compared
with samples packed in MAP. When the vacuum packages were
opened, the DeoxyMb was still able to bloom, OxyMb with a bright
red color was formed. This could be the reason why the a* value
increased in VP and VSP packed samples during the first two days in
air (Brewer, Zhu, Bidner, Meisinger, & McKeith, 2001). After 5 days
in dark aerobic storage, samples packed in VSP had the same redness
(a* value) as in MAP, but the redness of samples packed in VP was
significantly lower than in VSP. Furthermore, analogous tendencies
were observed on b* values in MAP, VP and VSP after 5 days in air.
These results suggest that VSP is better than VP regarding color stability.
The reason for differences in color stability between VP and VSP might
be attributed to different vacuum conditions during the packaging
process. Autoxidation of DeoxyMb to MetMb is very fast at low partial
oxygen pressure (George & Stratmann, 1952). The VSP had a heated
upper cover film that was shrunken tightly around the meat, which
probably reduced the possibility for residual oxygen to penetrate the
surface compared with VP where residual oxygen might penetrate the
meat surface and MetMb can be formed.
The relative contents of DeoxyMb, OxyMb and MetMb in meat de-
pend on the oxygen availability, the autoxidation rate of myoglobin
and the MetMb reducing capacity (Mancini & Hunt, 2005). The rela-
tive contents of DeoxyMb were higher in VP and VSP than in MAP
on day 0, which could be attributed to lower contents of oxygen in
VP and VSP. However, VSP has an advantage of minimized wrinkles
in the package compared with VP, which may decrease the speed
of bacterial growth (Vázquez et al., 2004). Barros-Velázquez et al.
(2003) found that aerobic mesophiles and lactic acid bacteria grew
more slowly in VSP than in VP. They also found that VSP prevented
the appearance of undesirable coloration and significantly improved

Table 2
Effect of packaging method (PM) and aerobic storage time (day) on α-tocopherol and NADH contents in meat.

PM Aging Number Day SE P-value

(day)
0 5 PM Day PM × Day

α‐Tocopherol (μg/g) Control 7 10 2.58aAB 2.16b 0.12 0.058 b0.001 0.044

MAP 14 8⁎ 2.27B 1.99 0.13
VP 14 10 2.46aAB 2.23b 0.12
VSP 14 5 2.59aA 2.00b 0.11
NADH (nmol/g) Control 7 10 114.23 118.54 4.53 0.004 0.002 0.719
MAP 14 8⁎ 115.09 121.52 4.25
VP 14 10 101.67 111.43 3.70
VSP 14 5 101.24b 113.06a 3.61

MAP: modified atmosphere packaging, 80% O2 + 20% CO2; VP: vacuum packaging; VSP: vacuum skin packaging. SE: standard error. Means with different low-case superscripts
within the same row (effect of aerobic storage time) differ at P b 0.05. Means with different capital superscripts within the same column (effect of PM) of the same trait differ at
P b 0.05 after Bonferroni correction. Number of comparisons = 6. ⁎One sample on day 5 with outlier value was not included in the result.
608 X. Li et al. / Meat Science 92 (2012) 604–609
the commercial color of the meat. The reduced risk of bacterial
growth may be beneficial for color stability. This may explain why
OxyMb was more stable in VSP than in VP during storage, resulting
in higher OxyMb and lower MetMb in VSP than in VP at the end of
storage. Differences in the relative contents of MetMb and DeoxyMb
between the different vacuum packed samples, control versus VP
and VSP could be attributed to different aging times. The whole LLs
were packed in vacuum at 1 day post mortem and stored for 7 days,
where after they were sliced and repacked for the different conditions.
The control samples were, at 7 days post mortem exposed to air,
whereas the other samples were stored for another 7 days before
exposure to air. Lindahl (2011) found that beef cut 2 days post mortem
was more unstable in color than beef cut after vacuum aging for 5
to 15 days. The authors attributed the higher oxidation to MetMb
and lower color stability to lower blooming ability at shorter aging
time. This early exposure to air could thus explain the higher relative
content of MetMb in the control compared with VP and VSP.
Lagerstedt, Lundström, et al. (2011) found that α-tocopherol con-
tent did not change with aging in vacuum, but decreased when the
beef was packed in vacuum for 5 days and then in MAP for 5 or
10 days. In the present study, α-tocopherol content showed the
largest decrease in VSP (22.8%) during storage compared with the
other methods. After 5 days of storage in air, the relative content of
OxyMb in VSP was slightly higher compared with VP. The mechanism
of α-tocopherol protecting oxymyoglobin is not well understood.
A model was proposed with a general hypothesis that α-tocopherol
indirectly maintained oxymyoglobin by directly inhibiting lipid
oxidation (Schaefer, Liu, Faustman, & Yin, 1995). The present results
showed that α-tocopherol was consumed during storage to maintain
the level of OxyMb confirming that α-tocopherol might influence
color stability (Faustman, Chan, Schaefer, & Havens, 1998).
The concentration of total NAD (NAD + and NADH) decreased
during storage post mortem in a study by Atkinson, Follett, and
Ratcliff (1969), whereas Giddings (1974) found that NADH could be
regenerated through reversal electron transport in mitochondria.
The regeneration of NADH post mortem is critical for delaying dis-
coloration (Ramanathan et al., 2011). As time post mortem proceeds,
some compounds could undergo gradual degradation and thus affect the
possibility of reversal electron transport. Andrews, Guthneck, Mcbride,
and Schweigert (1952) reported that adenosine triphosphatase, succinic
dehydrogenase and the glycolytic system showed no decrease in activity
in intact beef muscle after storage for two or four weeks at 2 °C. Thus,
NADH could be generated by these metabolic reactions post mortem.
In this study, the NADH concentration in VSP and VP increased after
display in air for 5 days, probably due to the regeneration of NADH
in the muscle. Pong, Chiou, Ho, and Jiang (2000) reported that NADH
concentration in tuna meat increased during the first 56–80 h of
storage, but was then followed by a rapid decrease. However, it is
established that NADH regeneration can occur with addition of sub-
strate post mortem. Kim et al. (2006) reported that beef strip loins
enhanced with 2.5% potassium lactate had significantly increased
color stability. They concluded that enhancing beef with lactate
replenished NADH via increased lactic dehydrogenase activity, resulting
in better meat color stability. In the present study, an increase in NADH
post mortem without any compound enhancement was observed. The
mechanism for this NADH increase post mortem is not known and
needs further study.
5. Conclusion
The results confirmed that packaging methods affect color stabili-
ty, as well as α-tocopherol and NADH contents. After vacuum aging,
samples of beef longissimus lumborum packed in VSP had better
color than those in VP after 5 days storage in air. The color of samples
packed in VSP was similar to those in MAP at the end (day 5) of
storage.
Acknowledgments
The authors thank China Scholarship Council for their financial
support. The authors would like to thank Åsa Lagerstedt Norström
and Maria Lundesjö Ahnström for their contributions to this project,
and also to Melvin C. Hunt from Kansas State University, USA for his
kind comments on the manuscript.
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