Meat Science 171 (2021) 108292

Contents lists available at ScienceDirect

Meat Science
journal homepage: www.elsevier.com/locate/meatsci

Effects of packaging methods combined with frozen temperature on the T

color of frozen beef rolls
Fangfang Wanga, Rongrong Lianga, Yimin Zhanga, Shujuan Gaob, Lixian Zhua, Lebao Niua,
Xin Luoa, Yanwei Maoa, , David L. Hopkinsa,c
⁎

a
College of Food Science and Engineering, Shandong Agricultural University, Taian, Shandong 271018, PR China
b
Animal Husbandry and Veterinary Service Centre of Daiyue District, Taian, Shandong 271000, PR China
c
Centre for Red Meat and Sheep Development, NSW Department of Primary Industries, Cowra, New South Wales 2794, Australia

ARTICLE INFO ABSTRACT

Keywords: Beef rolls for hot pot are usually stored and transported in a frozen state, and the beef color deteriorates quickly.
Freezing This paper reports on an investigation into the effect of packaging method, freezing temperature and storage
Shelf life time on instrumental color, pH, myoglobin state, lipid oxidation (TBARS) and total volatile basic nitrogen (TVB-
Myoglobin N) of beef rolls. It was shown that the color of beef rolls at −18 °C was better than that at −12 °C overall, and
Lipid oxidation
the OxyMb% and pH values were higher, while the MetMb% and TBARS were lower with storage at −18 °C.
Modified atmosphere packaging
With the extension of storage time, the instrumental color, OxyMb% and pH values of beef rolls decreased.
Correspondingly, the MetMb% and TBARS showed an upward trend. However, the TVB-N of all treatments did
not exceed the Chinese standard during 180d of storage. The results of this paper provide a number of re-
commendations for the storage of frozen beef rolls to extend color-shelf life.

1. Introduction such as iron, leading to an increase in reactive oxygen species and

Beef rolls are popular among Chinese consumers due to their tasti-
ness and high nutritional value, which makes them a widely used in-
gredient in hot pot meals. Beef rolls are usually stored and transported
in a frozen state, during which the color deteriorates quickly due to
freezing, an effect shown to be common for frozen meat (Cho et al.,
2017; Holman, Coombs, Morris, Kerr, & Hopkins, 2017). In addition,
previous research has shown that color is an important attribute that
influences consumers decisions to buy frozen beef rolls, and the a*
value (redness) is the most suitable index for predicting consumers'
color acceptance of frozen beef rolls. The acceptance threshold is a*
values ≥16.4 (Wang et al., 2020).
A number of studies have proposed that modified atmosphere
packaging (MAP) is one of the important methods to improve beef color
(Esmer, Irkin, Degirmencioglu, & Degirmencioglu, 2011; Hunt,
Mancini, Hachmeister, Kropf, & Milliken, 2004; Lindahl, 2011; Seyfert,
Hunt, Mancini, Hachmeister, & Unruh, 2004). However, these studies
have mainly focused on MAP used in chilled meat, and there has been
no research reported on the effect of MAP on frozen beef rolls. Com-
pared with chilled meat, the freezing process causes cell damage due to
the formation of ice crystals, and concentrates the co-oxidant solutes
oxidation reactions, which in turn affects the color stability of the meat
(Chakanya, Arnaud, Muchenje, & Hoffman, 2017). Therefore, in order
to extend the color-shelf life of frozen beef rolls, it is necessary to ex-
plore the proportion of gas in oxygen-containing packaging, especially
the oxygen content.
Now, −18 °C has been widely adopted as the standard operational
temperature for holding meat including in China, and − 12 °C as the
minimum threshold for frozen beef as per European Union definitions
(Anonymous, 2013). Some studies have indicated that beef stored at
−12 °C can maintain the same quality and shelf life as beef stored at
−18 °C (Holman et al., 2017; Qian et al., 2018). Thus, given potential
energy savings it is appropriate to examine the quality of beef rolls held
under MAP at the two different temperatures of −18 °C and − 12 °C.
Therefore, the objective of the present work was to optimize the
combination of different packaging methods (50% O2 + 30%
CO2 + 20% N2, 60% O2 + 40% N2, 80% O2 + 20% CO2, 0.4%
CO + 30% CO2 + 69.6% N2, overwrap packaging) and storage tem-
peratures (−12 °C and − 18 °C) to extend the color-shelf life of frozen
beef rolls held for up to 180 days.

⁎
Corresponding author.
E-mail address: maoyanwei@163.com (Y. Mao).

https://doi.org/10.1016/j.meatsci.2020.108292
Received 6 May 2020; Received in revised form 24 August 2020; Accepted 24 August 2020
Available online 02 September 2020
0309-1740/ © 2020 Elsevier Ltd. All rights reserved.
F. Wang, et al.
2. Materials and methods
2.1. Samples and treatments
Samples were obtained from the Longissmus Lumborum (LL) muscle
of six Chinese Luxi yellow cattle aged 18–24 months and weighing
between 560 and 600 kgs in a commercial abattoir. Carcasses were
conventionally chilled at 2–4 °C for 48 h. Both the left and right loins
were obtained at 48 h and vacuum packaged, and then transported to
the laboratory on ice in foam boxes, within 2 h. The LL were frozen at
−18 °C for 3 days. Before slicing, the LL were put into a flow tap water
bath for about half an hour and then each LL was cut into thin slices of
0.1 cm and rolled into solid beef rolls with a diameter of about 2 cm.
The slices were assigned randomly to ten package × temperature
groups (50% O2 + 30% CO2 + 20% N2; 60% O2 + 40% N2; 80%
O2 + 20% CO2; 0.4% CO + 30% CO2 + 69.6% N2 and overwrap
packaging, stored at −12 °C or − 18 °C). For each group there were 6
trays and each tray contained 10–12 beef rolls with a total weight of
about 120 g. Beef rolls were placed in polypropylene trays type TQBC-
0775 (oxygen transmission rate: 10 cm3/m2/24 h at 23 °C/0% relative
humidity, water vapor transmission rate: 15 g/m2/24 h at 38 °C/90%
relative humidity; Sealed Air Corp., Danbury, USA). Trays were then
flushed with the desired gas mixture and sealed with oxygen-barrier
film (oxygen transmission rate: 25 cm3/m2/24 h at 23 °C/0% relative
humidity, water vapor transmission rate: 10 g/m2/24 h at 4 °C/100%
relative humidity; Lid 1050, Sealed Air Corp., Danbury, USA) using a
DT-6D packaging machine (Dajiang Machinery Equipment Co., Ltd.,
Wenzhou, China). Carbon monoxide MAP was flushed with a certified
gas blend according to the combinations outlined previously (Xieli
Special Gas Co., Ltd., Jining, China). Overwrap packaging was sealed
with gas-permeable film (oxygen transmission rate: 16654 cm3/m2/
24 h/atm, carbon dioxide transmission rate: 64637 cm3/m2/24 h/atm,
water vapor transmission rate: 23.5 g/m2/24 h; QM/6105,
Qingqingmei Household Products Co., Ltd., Zhejiang, China). Each
treatment combination had six replicates (packages). The entire storage
period was in the dark and up to 180 days, with sampling on days 0, 30,
90, 120 and 180. The frozen temperatures were stable, and each sample
was held at its assigned temperature throughout the duration of the
storage period. Color, pH, relative levels of three myoglobin forms,
thiobarbituric acid reactive substances (TBARS) and total volatile basic
nitrogen (TVB-N) were determined. Samples used to determine relative
levels of three myoglobin forms, TBARS and TVB-N were stored at
−80 °C until analyzed.
2.2. Relative levels of myoglobin in three states measurement
Relative levels of myoglobin in three different forms was de-
termined according to Tang, Faustman, and Hoagland (2004) with
slight modification. Samples (4 g) were homogenized in 20 mL of
phosphate buffer solution (0.04 mol/L and pH 6.8) using an Ultra-
Turrax homogenizer (T18, IKA, Germany). After 1 h on the ice, the
homogenate was centrifuged at 4500 r/min for 30 min at 4 °C. The
supernatant was filtered through Whatman No. 1 filter paper. The fil-
tered fluid was scanned on an ultraviolet-visible spectrophotometer at
wavelengths of 582, 557, 525, and 503 nm, respectively, with the
phosphate buffer solution as a reference. The equations for determining
the level of each myoglobin form were as follows:
DeoMb% = 0.543 R1 + 1.594 R2 + 0.552 R3–1.329
OxyMb% + COMb% = 0.722 R1–1.432 R2–1.659 R3 + 2.599
MetMb% = 0.159 R1 0.085 R2 + 1.262 R3 0.520
where, R1 = A582/A525, R2 = A557/A525, R3 = A503/A525.
2
Meat Science 171 (2021) 108292
2.3. Color measurement
The surface color of beef rolls was determined by using an X-Rite
spectrophotometer (Model SP62, 4 mm diameter aperture, Illuminant
D65, 10o observer, X-Rite, USA). The first measurement was carried out
after blooming for 30 min at day 0, and the other measurements were
taken immediately after opening packages at days 30, 90, 120 and 180.
Six scans per roll were taken and CIE lightness (L*), redness (a*) and
yellowness (b*) were determined.
2.4. Lipid oxidation measurement
The lipid oxidation was determined by measurement of thiobarbi-
turic acid reactive substances (TBARS) using the method described by
Siu and Draper (1978) with slight modification. Samples (4 g) were
homogenized in 20 mL of distilled water using an Ultra-Turrax T18
homogenizer (T18; IKA, Germany) for 1 min. Subsequently, 20 mL of
10% (w/v) trichloroacetic acid (TCA) was added and the mixture was
vortexed, and then filtered through Whatman No. 1 filter paper. Then,
1 mL of the filtrate was taken and added to 250 μL of 60 mM 2-thio-
barbituric acid (TBA). Subsequently, the solution was incubated at
80 °C for 90 min. After that, the absorbance of the solution was mea-
sured using a Microplate reader (Epoch TM 2, Bio Tek, America) at
532 nm against a blank containing all reagents. Results were calculated
from a standard curve and expressed as TBARS in mg malondialdehyde
(MDA) per kg sample.
2.5. pH values measurement
Meat samples of about 3 g without tendon and fat were placed in
50 mL centrifuge tubes and 30 mL of deionized water was added. The
samples were homogenized for 30s with an Ultra-Turrax homogenizer
(T18, IKA, Germany) until the samples were fully mixed, but not
emulsified. The pH of samples was measured using a pH meter (S210,
Mettler, Switzerland). The pH electrode was calibrated with two buffers
(pH 4.0 and 7.0) and the temperature was below 25 °C during the whole
measurement process. Each sample was measured six times and the
results averaged.
2.6. TVB-N measurement
The TVBN content was determined using a Kjeldahl method as de-
scribed by the Chinese National Food safety standard methods GB
5009.228–2016. Samples (10 g) were dispersed in 75 mL of distilled
water and equilibrated for 30 min. Then, the equilibrated solution was
poured into the distillation tube and 1 g of magnesium oxide added. The
distillation tube was immediately connected to the distiller of a Kjeldahl
nitrogen analyzer and distilled for 180 s and the distillate was mixed
with 20 g/L boric acid solution. The TVB-N values were determined
according to the consumption of hydrochloric acid and calculated using
the following equation:
TVBN (mg/100 g) = ((V1 V2) × c × 14)/m × 100
where: V1 denotes the volume of the hydrochloric acid (mL) con-
sumed by a tested sample. V2 denotes the volume of the hydrochloric
acid (mL) consumed by a blank sample. c denotes the actual con-
centration of hydrochloric acid (mol/L). 14 denotes molar mass of ni-
trogen. m denotes sample weight (the unit is gram).
2.7. Statistical analysis
The MIXED procedure (SAS, Version 9.0) was used with packaging
method, storage temperature, storage time and their interaction as fixed
factors, and carcass as a random factor. Least square means were se-
parated using the PDIFF option and were considered significant at
P < 0.05.
F. Wang, et al.
Table 1
Effect of different storage temperature and time on DeoMb% levels of beef rolls during storage.
Storage temperature (°C) Storage time (days)
0 30
ax
−12 14.22 ± 0.25 12.80 ± 0.40
−18 14.22 ± 0.25ax 12.89 ± 0.38bx
Note: a-c Different letters indicate that there were significant differences in different storage times for the same storage temperature, P < 0.05;
indicate that there were significant differences between different storage temperatures for the same storage time, P < 0.05.
3. Results and discussion
3.1. Relative levels of myoglobin forms
Myoglobin in beef usually exists in three forms, as deoxymyoglobin
(DeoMb), oxygenated myoglobin (OxyMb) and metmyoglobin (MetMb)
respectively and the relative levels of the three forms is one of the
important factors influencing beef meat color (Ramanathan, Nair, Hunt,
& Suman, 2019). But it is worth noting that, OxyMb has two absorbance
peaks at 542 and 582 nm (Tang et al., 2004) and carboxymyoglobin
(COMb) has two absorbance peaks at 543 and 581 nm (Suman,
Mancini, & Faustman, 2006). Hence it is very difficult to differentiate
these two Mb forms using absorption spectrum and the oxygenated
myoglobin in samples held under CO MAP in this study probably also
represented any COMb that was formed.
3.1.1. DeoMb%
There was a storage temperature × storage time interaction for
DeoMb% of beef rolls (P < 0.05; Table 1) such that after extended
storage time samples stored at −18 °C exhibited lower DeoMb% levels
than if stored at −12 °C. DeoMb% levels decreased significantly as
storage time increased and reached an earlier plateau in samples stored
at −12 °C (P < 0.05).
It can be seen from Fig. 1 that packaging method had an effect on
the DeoMb% of beef rolls (P < 0.05). The DeoMb% of beef rolls in CO
MAP was higher than that of the other four packaging methods
(P < 0.05).
Fig. 1. Effect of packaging method on DeoMb% of beef rolls. Packaging
methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for
60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for 0.4%
CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-b Different
letters indicated that there were significant differences in DeoMb% between
different packaging methods, P < 0.05.
Meat Science 171 (2021) 108292
90 120 180
bx cy cx
11.60 ± 0.21 11.57 ± 0.30 11.05 ± 0.56cx
12.71 ± 0.33bx 11.55 ± 0.31cx 10.20 ± 0.35dy
x-y
Different letters
3.1.2. OxyMb%
The interaction between storage time and packaging method had an
effect on OxyMb% (P < 0.05) as shown in Table 2. The OxyMb% level
did not change for 30 days (P > 0.05) for any of the five packaging
methods, but OxyMb% of the beef rolls in 80% O2 MAP and overwrap
packaging after 90 days was significantly lower than or not significantly
higher than the other three packaging methods (P < 0.05). With the
prolongation of storage time, the OxyMb% level of the beef rolls in all
packaging methods except CO MAP showed a significant decline up to
90 days. However, after 90 days, the OxyMb% in the CO MAP treatment
increased, whereas the overwrap packaging showed a significant in-
crease between 120 and 180 days. These changes were linked to a re-
duction in MetMb% indicating a reversal in oxygen binding, an ob-
servation which requires further investigation. There was also an
interaction of storage temperature × packaging method for OxyMb%
(P < 0.05; Table 3). The OxyMb% of beef rolls in CO MAP at −12 °C
was significantly higher or not significantly lower than that of other
packaging methods, while the OxyMb% of beef rolls in 5 packaging
methods at −18 °C showed no significant change (P < 0.05). The
OxyMb% in beef rolls at −18 °C was significantly higher or not sig-
nificantly lower than −12 °C except for CO MAP (P < 0.05).
Results shown in Table 4 indicated that the storage temperature ×
storage time interaction had an effect on OxyMb% (P < 0.05). The
OxyMb% in beef rolls decreased significantly at −12 °C over the first
90 days of storage time, but then exhibited no change while the OxyMb
% of beef rolls at −18 °C exhibited small inconsistent changes, with no
difference between values at 0 and 180d.
3.1.3. MetMb%
It can be seen from Table 5 that there was an interaction (P < 0.05)
between storage time and packaging method on the MetMb% values.
There was no significant change in the MetMb% of the beef rolls be-
tween the 5 packaging methods for 30 days, while the MetMb% of the
beef rolls in the 50% O2 MAP, 80% O2 MAP and overwrap packaging
after 90 days was significantly higher or not significantly lower than the
other two packaging methods. With the prolongation of storage time,
the MetMb% of beef rolls in other packaging methods except CO MAP
showed a significant increase.
An interaction of storage temperature and packaging method oc-
curred for MetMb% of beef rolls (P < 0.05) as shown in Table 6. The
MetMb% of beef rolls in 50% O2 MAP and 80% O2 MAP at −12 °C was
significantly higher or not significantly lower than the other packaging
methods, while the MetMb% in five packaging methods at −18 °C
showed no significant change. The MetMb% in beef rolls at −18 °C was
significantly lower or not significantly higher than −12 °C except for
CO MAP. The change trend of the MetMb% was basically opposite to
that of the OxyMb%.
The relative levels of myoglobin in three states is one of the im-
portant indicators reflecting beef meat color, especially OxyMb% and
MetMb% and it is closely related to a* values (Bekhit, Morton, Bhat, &
Kong, 2019). Oxygenated myoglobin in beef is bright cherry red, but
metmyoglobin is brown, so the higher the OxyMb%, the redder the
meat color. In this study, the trend of the effect of packaging method,
storage temperature and storage time on the OxyMb% and MetMb%
was similar to a* values (as shown in Table 7). At −12 °C, the
3
F. Wang, et al. Meat Science 171 (2021) 108292

Table 2
Effect of different packaging methods on OxyMb% of beef rolls during storage.
Storage time (days) Packaging method

50%O2 60%O2 80%O2 CO OP

ax ax ax ay
0 50.13 ± 1.70 50.13 ± 1.70 50.13 ± 1.70 50.13 ± 1.70 50.13 ± 1.70ax
30 46.75 ± 2.35axy 49.04 ± 2.22ax 47.12 ± 3.24axy 49.05 ± 2.18ay 48.91 ± 2.09ax
90 43.09 ± 2.76byz 46.90 ± 1.27abx 37.25 ± 3.31cz 49.46 ± 2.33ay 42.98 ± 2.81byz
120 42.86 ± 3.70bcyz 44.91 ± 3.32bx 41.22 ± 3.11bcz 57.14 ± 2.45ax 38.78 ± 2.69cz
180 40.69 ± 3.53cz 47.58 ± 2.01bx 42.17 ± 3.11bcyz 58.26 ± 2.34ax 45.94 ± 2.15bcxy

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-c Different letters indicated that there were significant differences between different packaging
methods for the same storage time, P < 0.05; w-z Different letters indicated that there were significant differences between different storage times for the same
packaging method, P < 0.05.

packaging method with the highest a* values were CO MAP, followed
by 60% O2 MAP and the worst was 80% O2 MAP (as shown in Table 8).
The OxyMb% in CO MAP was the highest, followed by 60% O2 MAP,
and 80% O2 MAP was the worst, while the MetMb% was opposite to
that of OxyMb% (as shown in Tables 3 and 6). There was no significant
difference in a* values, OxyMb% and MetMb% of five packaging
methods at −18 °C (as shown in Tables 3, 6 and 8). Except for CO MAP,
the a* values and the OxyMb% in beef rolls at −18 °C were sig-
nificantly higher than those at −12 °C, while the MetMb% was opposite
(as shown in Tables 3, 6 and 8). With the extension of storage time, a*
values and OxyMb% of beef rolls were mostly significantly decreased,
while the MetMb% was opposite (as shown in Tables 2, 5 and 7).
In addition, it is well known that the three forms of myoglobin can
be converted into each other through redox reactions (Bekhit et al.,
2019; Gao, Wang, Tang, & Dai, 2014). This redox reaction is greatly
affected by temperature. For example, the oxidation rate of DeoMb and
OxyMb to MetMb was faster at a higher storage temperature, and the
lower the storage temperature, the slower the oxidation rate of myo-
globin (Choe, Stuart, & Kim, 2016).
3.2. Color
higher temperature (−12 °C), which was contrary to the other four
packaging methods in which it was apparent that the colder the tem-
perature of storage the better the retention of redness (higher a* values)
(Table 8). The reduction of a* values was associated with the oxidation
of myoglobin (especially OxyMb) to form MetMb, which is accelerated
by high temperature (O'Keeffe & Hood, 1982; Renerre, 1999). In ad-
dition, lipid oxidation is also significantly correlated with myoglobin
oxidation (Faustman, Sun, Mancini, & Suman, 2010) while low tem-
perature can inhibit lipid oxidation to a certain extent (Lan, Shang,
Song, & Dong, 2016). However, the mechanism of the influence of
temperature on CO MAP is worth further study.
3.2.2. L* values
Only the storage time had an effect on the L* values of beef rolls
(P < 0.05). This result is consistent with the study of Esmer et al.
(2011), who found that the packaging method had no impact on L*
values of ground beef, while the L* values were significantly affected by
the storage time. It can be seen from Fig. 2 that with the extension of
storage time, the L* values of beef rolls generally showed a trend of
decreasing first and then increasing such that the beef rolls frozen for
30d had the lowest L* values.

3.2.1. a* values 3.2.3. b* values

The data analysis showed that the interaction between storage time
and packaging method (Table 7) and storage temperature and packa-
ging method (Table 8) had an effect on a* values of the beef rolls
(P < 0.05).
With the extension of storage time, the a* values of the beef rolls in
all the five packaging methods showed a significant decline, but the
change in the CO MAP was small (Table 7), as the CO MAP would be
low in O2, and the stability of carbonyl myoglobin is higher so the a*
values decreased slowly (Zhang et al., 2017). Related to this Kang,
Kang, Seong, Park, and Cho (2014) found that a* values of beef slices in
oxygen-containing packaging decreased significantly with the extension
of storage time and Lindahl (2011) found that the meat color of beef
steaks in high‑oxygen packaging deteriorated with storage time.
For the CO MAP packaged samples, the lower storage temperature
(−18 °C) lead to a decrease of a* values (P < 0.05) compared to the
The results showed that only the storage time (Fig. 3 (A)) and
packaging method (Fig. 3 (B)) had an effect on the b* values
(P < 0.05). It can be seen from Fig. 3 (A) that with the extension of
storage time, the b* values of beef rolls showed a significant downward
trend overall (P < 0.05). It was found that the b* values were sig-
nificantly correlated with a* values (Esmer et al., 2011), which meant
that the meat color deteriorated when a* values decreased, and b*
values decreased due to the formation of metmyoglobin, which changed
the color to browny red. During storage, the b* values of beef rolls in
80% O2 and CO MAP were significantly lower or not significantly
higher than other packaging methods (P < 0.05; Fig. 3 (B)).
Combined with the color acceptance threshold (a* ≥ 16.4) obtained
from previous work (Wang et al., 2020) and the results in Table 9, and
also from the comprehensive consideration of economic factors, the
following conclusions are drawn. The optimal packaging method for

Table 3
Effect of different packaging methods on OxyMb% values of beef rolls at two storage temperatures.
Storage temperature (°C) Packaging method

50%O2 60%O2 80%O2 CO OP

cdy bx dy ax
−12 40.28 ± 2.13 46.17 ± 1.59 37.85 ± 2.19 56.50 ± 1.36 43.12 ± 1.96bcy
−18 49.13 ± 1.13ax 49.25 ± 1.11ax 49.31 ± 1.00ax 49.11 ± 1.43ay 47.57 ± 1.04ax

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-d Different letters indicated that there were significant differences between different packaging
methods for the same storage temperature, P < 0.05; x-y Different letters indicated that there were significant differences between different storage temperatures for
the same packaging method, P < 0.05.

4
F. Wang, et al. Meat Science 171 (2021) 108292

Table 4
Effect of different storage temperatures on OxyMb% values of beef rolls during storage.
Storage temperature (°C) Storage time (days)

0 30 90 120 180

ax bx cx cy
−12 50.13 ± 1.05 46.51 ± 1.69 42.41 ± 1.91 41.10 ± 2.19 43.78 ± 1.97bcy
−18 50.13 ± 1.05ax 49.83 ± 1.27ax 45.46 ± 1.54bx 48.87 ± 2.15abx 50.08 ± 1.64ax

Note: a-c Different letters indicated that there were significant differences between different storage times for the same storage temperature, P < 0.05; x-y Different
letters indicated that there were significant differences between different storage temperatures for the same storage time, P < 0.05.

Table 5
Effect of different packaging methods on MetMb% of beef rolls during storage.
Storage time (days) Packaging method

50%O2 60%O2 80%O2 CO OP

az az az ax
0 35.79 ± 1.39 35.79 ± 1.39 35.79 ± 1.39 35.79 ± 1.39 35.79 ± 1.39az
30 40.26 ± 1.98ayz 37.85 ± 1.67ayz 41.23 ± 2.92ay 37.36 ± 1.97ax 38.82 ± 2.81az
90 44.88 ± 2.82bxy 41.08 ± 0.74bcxy 50.61 ± 3.43ax 37.73 ± 2.28cx 45.53 ± 2.65bxy
120 45.90 ± 3.57abx 43.45 ± 3.09bx 47.45 ± 2.93abx 30.64 ± 2.27cy 50.42 ± 2.46ax
180 48.77 ± 3.39ax 42.75 ± 1.89bxy 46.70 ± 2.31abx 30.28 ± 2.20cy 44.01 ± 1.65aby

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-c Different letters indicated that there were significant differences between different packaging
methods for the same storage time, P < 0.05; x-z Different letters indicated that there were significant differences between different storage times for the same
packaging method, P < 0.05.

Table 6
Effect of different packaging methods on MetMb% values of beef rolls at two storage temperatures.
Storage temperature (°C) Packaging method

50%O2 60%O2 80%O2 CO OP

−12 47.54 ± 2.22abx 42.05 ± 1.56cx 49.86 ± 2.13ax 30.65 ± 1.13dy 45.20 ± 1.91bcx
−18 38.71 ± 0.97ay 38.32 ± 0.84ay 38.85 ± 0.87ay 38.07 ± 1.30ax 40.63 ± 1.00ay

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-c Different letters indicated that there were significant differences in different packaging
methods for the same storage temperature, P < 0.05; x-y Different letters indicated that there were significant differences between different storage temperatures for
the same packaging method, P < 0.05.

Table 7
Effect of different packaging methods on a* values of beef rolls during storage.
Storage time (days) Packaging method

50%O2 60%O2 80%O2 CO OP

ax ax ax ax
0 22.27 ± 0.26 22.27 ± 0.26 22.27 ± 0.26 22.27 ± 0.26 22.27 ± 0.26ax
30 20.09 ± 0.68ax 20.70 ± 0.54axy 18.42 ± 1.06aby 17.32 ± 1.04bz 19.44 ± 1.09aby
90 17.08 ± 1.13aby 18.68 ± 0.86ay 14.03 ± 1.46cz 19.26 ± 1.72ayz 15.69 ± 1.02bcz
120 13.60 ± 1.28bcz 15.87 ± 0.88bz 12.31 ± 1.59cz 20.05 ± 1.58axy 13.55 ± 1.19bcz
180 13.45 ± 1.22cz 16.12 ± 0.99bz 13.89 ± 1.39bcz 19.20 ± 1.33ayz 14.00 ± 1.04bcz

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-c Different letters indicated that there were significant differences between different packaging
methods at the same storage time, P < 0.05; w-z Different letters indicated that there were significant differences between different storage times for the same
packaging method, P < 0.05.

Table 8
Effect of different packaging methods on a* values of beef rolls at two storage temperatures.
Storage temperature (°C) Packaging method

50%O2 60%O2 80%O2 CO OP

cy bx dy ax
−12 16.19 ± 1.06 18.04 ± 0.76 13.57 ± 1.11 20.88 ± 0.89 15.89 ± 0.97cy
−18 18.41 ± 0.62ax 19.42 ± 0.50ax 18.80 ± 0.63ax 18.36 ± 0.74ay 18.09 ± 0.70ax

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-d Different letters indicated that there were significant differences between different packaging
methods at the same storage temperature, P < 0.05; x-y Different letters indicated that there were significant differences between different storage temperatures
using the same packaging method, P < 0.05.

5
F. Wang, et al. Meat Science 171 (2021) 108292

a-d
Fig. 2. Changes in L* values of beef rolls during storage times. Different letters indicated that there were significant differences in L* values between different
storage times, P < 0.05.

Fig. 3. Effects of storage time (A) and packing method (B) on b* values of beef rolls. Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2
short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for 0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-d Different
letters indicated that there were significant differences in b* values between different storage times, P < 0.05; x-z Different letters indicated that there were
significant differences in b* values between different packaging methods, P < 0.05.

Table 9
Effects of different packaging methods on a* values of beef rolls at different temperatures during storage.
Storage temperature (°C) Storage time (days) Packaging method

50%O2 60%O2 80%O2 CO OP

akx akx akx akx

−12 0 22.27 ± 0.38 22.27 ± 0.38 22.27 ± 0.38 22.27 ± 0.38 22.27 ± 0.38akx
30 19.89 ± 1.02akx 20.05 ± 0.74aklx 15.67 ± 0.98bly 17.81 ± 1.53ablx 18.73 ± 1.38ablx
90 15.82 ± 1.94bclx 17.98 ± 1.52ablmx 9.96 ± 1.42dmy 20.88 ± 2.60aklx 14.67 ± 1.84cmx
120 11.81 ± 2.04bcmy 14.84 ± 1.44bmx 9.07 ± 1.97cmy 23.05 ± 2.20akx 11.28 ± 1.40cny
180 11.14 ± 2.07cmy 15.05 ± 1.86bmx 10.87 ± 2.16cmy 20.39 ± 2.39aklx 12.49 ± 1.76bcmnx
−18 0 22.27 ± 0.38akx 22.27 ± 0.38akx 22.27 ± 0.38akx 22.27 ± 0.38akx 22.27 ± 0.38akx
30 20.29 ± 0.99aklx 21.36 ± 0.74akx 21.18 ± 0.96aklx 16.83 ± 1.53blx 20.15 ± 1.76aklx
90 18.33 ± 1.10almx 19.39 ± 0.86aklx 18.11 ± 0.84almx 17.64 ± 2.29alx 16.70 ± 0.90almx
120 15.39 ± 1.32amx 16.90 ± 0.96alx 15.55 ± 1.76amx 17.04 ± 1.57aly 15.83 ± 1.49amx
180 15.75 ± 0.43amx 17.19 ± 0.61alx 16.90 ± 0.49amx 18.01 ± 1.23alx 15.50 ± 0.87amx

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-d Different letters indicate significant differences between different packaging methods at the
same temperature and storage time, P < 0.05; k-n Different letters indicate significant differences between different storage times at the same temperature and
packaging method, P < 0.05; x-y Different letters indicate significant differences between different storage temperatures under the same storage time and packaging
method, P < 0.05.

frozen beef rolls stored for 30 days is overwrap packaging at −12 °C.
For countries where CO MAP is forbidden, the optimal packaging
method is 60% O2 MAP when stored at −12 °C for more than 30 to
90 days. For countries where CO MAP is allowed, the optimal packa-
ging method is CO MAP when stored for more than 30 to 180 days at
−12 °C. In addition, at −18 °C, the most suitable packaging method for
frozen beef rolls when stored to 90 days is overwrap packaging, and the
most suitable packaging method for storage from 90 to 180 days is 60%
O2 MAP or CO MAP (for countries where CO MAP is allowed) and in
general storage at the lower temperature is better.

6
F. Wang, et al. Meat Science 171 (2021) 108292

Table 10
Effect of different packaging methods on TBARS (mg MDA/ kg) of beef rolls during storage.
Storage time (days) Packaging method

50%O2 60%O2 80%O2 CO OP

az ay ay aw
0 0.11 ± 0.01 0.11 ± 0.01 0.11 ± 0.01 0.11 ± 0.01 0.11 ± 0.01az
30 0.29 ± 0.05byz 0.29 ± 0.06bxy 0.71 ± 0.21ax 0.17 ± 0.02bw 0.39 ± 0.12abyz
90 0.60 ± 0.16bcxy 0.45 ± 0.08cwxy 0.97 ± 0.23awx 0.22 ± 0.03cw 0.92 ± 0.43abwx
120 0.71 ± 0.18bwx 0.66 ± 0.14bwx 1.19 ± 0.21aw 0.27 ± 0.03cw 1.21 ± 0.32aw
180 1.04 ± 0.23aw 0.71 ± 0.17aw 1.04 ± 0.21awx 0.22 ± 0.01bw 0.74 ± 0.22axy

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-c Different letters indicated that there were significant differences between different packaging
methods for the same storage time, P < 0.05; w-z Different letters indicated that there were significant differences between different storage times for the same
packaging method, P < 0.05.

Table 11
Effect of different packaging methods on TBARS (mg MDA/ kg) values of beef rolls at two storage temperatures.
Storage temperature (°C) Packaging method

50%O2 60%O2 80%O2 CO OP

bcx cx ax dx
−12 0.74 ± 0.13 0.63 ± 0.10 1.21 ± 0.15 0.23 ± 0.02 0.97 ± 0.21bx
−18 0.36 ± 0.08ay 0.26 ± 0.03ay 0.40 ± 0.07ay 0.17 ± 0.01ax 0.38 ± 0.11ay

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging; a-d Different letters indicated that there were significant differences between different packaging
methods for the same storage temperature, P < 0.05; x-y Different letters indicated that there were significant differences between different storage temperatures for
the same packaging method, P < 0.05.

Fig. 4. Effects of storage time (A), packaging method (B) and storage temperature (C) on pH values of beef roll. Packaging methods: 50% O2 short for 50% O2 + 30%
CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for 0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap
packaging; a-c Different letters indicate that there are significant differences in pH values at different storage times, P < 0.05; l-n Different letters indicated that there
were significant difference in pH values between different packaging methods, P < 0.05; x-y Different letters indicated that there were significant differences in pH
values between different storage temperatures, P < 0.05.

Table 12
Effects of different packaging methods on TVB-N (mg/kg) of beef rolls at different temperatures during storage.
Storage temperature (°C) Storage time (days) Packaging method

50%O2 60%O2 80%O2 CO OP

−12 0 12.04 ± 0.23 12.04 ± 0.23 12.04 ± 0.23 12.04 ± 0.23 12.04 ± 0.23
180 12.79 ± 0.66 12.77 ± 0.72 13.18 ± 1.26 12.70 ± 0.81 12.92 ± 1.04
−18 0 12.04 ± 0.23 12.04 ± 0.23 12.04 ± 0.23 12.04 ± 0.23 12.04 ± 0.23
180 12.94 ± 0.75 12.27 ± 0.59 12.25 ± 0.82 12.12 ± 0.66 12.77 ± 0.54

Note: Packaging methods: 50% O2 short for 50% O2 + 30% CO2 + 20% N2, 60% O2 short for 60% O2 + 40% N2, 80% O2 short for 80% O2 + 20% CO2, CO short for
0.4% CO + 30% CO2 + 69.6% N2, OP short for overwrap packaging.

3.3. Lipid oxidation
The main products of lipid oxidation are peroxides, which in turn
decompose under the action of peroxidase to form low molecular
weight products, such as aldehydes (malondialdehyde), ketones, fatty
acids, etc. (Faustman et al., 2010). Among them, malondialdehyde can
react with thiobarbituric acid to form color, so this reaction (TBARS) is
used to measure the degree of lipid oxidation.
Storage time and packaging method interacted (P < 0.05) to affect
the TBARS as shown in Table 10. Except for day 0, the lipid oxidation of

7
F. Wang, et al.
the beef rolls in 80% O2 MAP was the highest, followed by overwrap
packaging. As storage time increased lipid oxidation of the beef rolls
increased for all packaging methods apart from that based on CO,
which showed only a small change. There is an international consensus
that oxygenated packaging promotes lipid oxidation (Cayuela & Gil,
2004; Kim, Huff-Lonergan, Sebranek, & Lonergan, 2010), and the
higher the oxygen content, the higher the degree of lipid oxidation
(Łopacka, Półtorak, & Wierzbicka, 2016; Esmer et al., 2011), but this
was not the case at the end of the storage time where MAP stored
samples containing oxygen had oxidized to the same extent as each
other and equivalent to overwrap.
An interaction of packaging method and storage temperature was
evident for TBARS (P < 0.05; Table 11). The lipid oxidation of beef
rolls in 80% O2 MAP at −12 °C was significantly higher than that of the
other packaging methods, while the lipid oxidation of beef rolls in 5
packaging methods at −18 °C was similar. Except for CO MAP, the lipid
oxidation of beef rolls in the remaining 4 packaging methods at −12 °C
was significantly higher than at −18 °C. CO MAP has certain anti-
oxidant properties, and the response reported here is consistent with
the results of Zhang et al. (2017) and Van Rooyen, Allen, and O'Connor
(2017) because the antioxidant capacity of carboxymyoglobin is
stronger than that of oxygenated myoglobin. Numerous studies have
shown that lipid oxidation represented by TBARS is significantly related
to the redox state of myoglobin (Baron & Andersen, 2002; Chaijan,
2008; Faustman et al., 2010). These studies defined myoglobin oxida-
tion and lipid oxidation as interactions, since the secondary oxidation
products released during the oxidation of one substance can accelerate
the reaction of the other. Our study also showed that the color of frozen
beef rolls at −18 °C was better than those stored at −12 °C (apart from
those in CO MAP), and that a lower temperature can inhibit lipid oxi-
dation to a certain extent, as also observed by Lan et al. (2016).
3.4. pH values
pH values are an important indicator of meat quality and an im-
portant determinant of microbial growth. The data analysis indicated
that the fixed factors of storage time, packaging method and storage
temperature had an effect on the pH values of beef rolls independently
of each other (P < 0.05).
According to Fig. 4 (A), storage time had a significant effect on the
pH values of beef rolls (P < 0.05). Overall, the pH values of beef rolls
decreased significantly with storage time. This is probably caused by
the denaturation of the buffer proteins and the increase of solute con-
centration due to the change of the frozen exudate (Leygonie, Britz, &
Hoffman, 2012). The results in Fig. 4 (B) indicate that the packaging
method had a significant effect on the pH values of beef rolls
(P < 0.05). The pH values of beef rolls in 50% O2 MAP and 60% O2
MAP were significantly higher or not significantly lower than the other
three packaging methods, while the pH values of 80% O2 MAP and
overwrap packaging were significantly lower or not significantly higher
than other packaging methods. As shown in Fig. 4 (C), the storage
temperature had an effect on the pH values of beef rolls (P < 0.05),
with the pH values of beef rolls at −18 °C being significantly higher
than at −12 °C.
Many studies have shown that pH is negatively correlated with lipid
oxidation, and the more severe the lipid oxidation, the lower the pH
(Guo et al., 2015), which may be caused by fatty acids and other acids
produced by lipid oxidation.
3.5. TVB-N
TVB-N is widely used as an indicator of shelf life and freshness for
fleshy foods (Cortez-Vega, Pizato, & Prentice, 2012; Liu, Li, Xia,
Regenstein, & Zhou, 2013). According to the data analysis, the factors
storage time, storage temperature and packaging method had no sig-
nificant effect on TVB-N levels of the beef rolls (P > 0.05). TVB-N is
8
Meat Science 171 (2021) 108292
mainly composed of ammonia, amine and trimethylamine, which ori-
ginate from microorganisms and enzyme activity (Lu, Tang, & Dong,
2017). As shown in Table 12, the TVB-N values of beef rolls increased
slightly at both temperatures during storage, and some studies have
shown that this is related to the increase in the total viable counts (TVC)
during storage (Kyrana, Lougovois, & Valsamis, 1997; Qian et al.,
2018). For commercial fresh and frozen meat products, the Chinese
National Food Safety Standard (GB 2707–2016) allows maximum TVB-
N values of 15 mg/100 g. The TVB-N values of all treatment groups in
this study did not exceed the limit during the storage period as showed
in Table 12. This result was in agreement with the research results of
Qian et al. (2018), who found that the TVB-N values of beef stored at
−12 °C and − 18 °C for 168 days did not exceed the national limit.
4. Conclusions
In general, CO MAP and 60% O2 + 40% N2 MAP provided better
color protection for frozen beef meat rolls, while 80% O2 + 20% CO2
MAP had the worst color protection effect. With the extension of sto-
rage time, the color of beef rolls showed a downward trend, and the
color of frozen beef rolls at −18 °C was better than at −12 °C. At the
same time, the trend in the variation of OxyMb% and pH values was
consistent with a* values, while the variation trend between MetMb%
and lipid oxidation are opposite. In conclusion, this study established
the best storage methods for frozen beef rolls in terms of color stability
which could improve product quality while reducing costs and creating
greater economic benefit. As follows: when stored at −12 °C and the
storage period is within 30 days, it is recommend to use overwrap
packaging, and when the storage period needs to reach 30–90 days,
60% O2 + 40% N2 MAP is recommended for countries where CO MAP
is forbidden, and when the storage period needs to reach 30–180 days,
CO MAP is recommended for countries where CO MAP is allowed.
When stored at −18 °C and the storage period is within 90 days,
overwrap packaging is recommended, and when the storage period
needs to reach 90–180 days, 60% O2 + 40% N2 MAP or CO MAP are
recommended for different countries.
Declaration of Competing Interest
The authors state that they have no conflict of interests.
Acknowledgements
This work was supported by the Ministry of Agriculture and Rural
Affairs of the People's Republic of China (CARS-37); Shandong
Provincial Department of Agriculture (SDAIT-09-09); Shandong
Agricultural University (SYL2017XTTD12).
References
Anonymous (2013). Opening and providing for the administration of tariff quotas for high-
quality fresh, chilled and frozen beef and for frozen buffalo meat. Commission im-
plementing regulation. BEL: European Union1–12.
Baron, C. P., & Andersen, H. J. (2002). Myoglobin-induced lipid oxidation. A review.
Journal of Agricultural and Food Chemistry, 50, 3887–3897.
Bekhit, E. D. A., Morton, J. D., Bhat, Z. F., & Kong, L. (2019). Meat color: Factors affecting
color stability. Encyclopedia of Food Chemistry, 2, 202–210.
Cayuela, J. M., & Gil, M. D. (2004). Effect of vacuum and modified atmosphere packaging
on the quality of pork loin. European Food Research and Technology, 219, 316–320.
Chaijan, M. (2008). Review: Lipid and myoglobin oxidations in muscle foods.
Songklanakarin Journal of Science and Technology, 30, 47–53.
Chakanya, C., Arnaud, E., Muchenje, V., & Hoffman, L. C. (2017). Colour and oxidative
stability of mince produced from fresh and frozen/thawed fallow deer (dama dama)
meat. Meat Science, 126, 63–72.
Cho, S., Kang, S. M., Seong, P., Kang, G., Kim, Y., Kim, J., ... Park, B. (2017). Effect of
aging and freezing conditions on meat quality and storage stability of 1++ grade
Hanwoo steer beef: Implications for shelf life. Korean Journal for Food Science of
Animal Resources, 37(3), 440–448.
Choe, J. H., Stuart, A., & Kim, Y. H. B. (2016). Effect of different aging temperatures prior
to freezing on meat quality attributes of frozen/thawed lamb loins. Meat Science, 116,
F. Wang, et al.
158–164.
Cortez-Vega, W. R., Pizato, S., & Prentice, C. (2012). Quality of raw chicken breast stored
at 5c and packaged under different modified atmospheres. Journal of Food Safety, 32,
360–368.
Esmer, O. K., Irkin, R., Degirmencioglu, N., & Degirmencioglu, A. (2011). The effects of
modified atmosphere gas composition on microbiological criteria, color and oxida-
tion values of minced beef meat. Meat Science, 88(2), 221–226.
Faustman, C., Sun, Q., Mancini, R., & Suman, S. P. (2010). Myoglobin and lipid oxidation
interactions: Mechanistic bases and control. Meat Science, 86(1), 86–94.
Gao, X. G., Wang, Z. Y., Tang, M. T., & Dai, R. T. (2014). Comparison of the effects of
succinate and NADH on postmortem metmyoglobin reductase activity and beef
colour stability. Journal of Integrative Agriculture, 13(8), 1817–1826.
Guo, J., Wang, J., Lin, H., Zhang, Y., Hu, H., Wang, C., & Wu, Y. (2015). Changes and
correlations of Duroc muscle pH, glycogen, lactic acid and TBA under different sto-
rage conditions. Agricultural Science and Technology, 16(12), 2795–2800.
Holman, B. W. B., Coombs, C. E. O., Morris, S., Kerr, M. J., & Hopkins, D. L. (2017). Effect
of long term chilled (up to 5 weeks) then frozen (up to 12 months) storage at two
different sub-zero holding temperatures on beef: 1. Meat quality and microbial loads.
Meat Science, 133, 133–142.
Hunt, M. C., Mancini, R. A., Hachmeister, K. A., Kropf, D. H., & Milliken, G. (2004).
Carbon monoxide in modified atmosphere packaging affects color, shelf life, and
microorganisms of beef steaks and ground beef. Journal of Food Science, 69(1),
FCT45–FCT52.
Kang, S. M., Kang, G., Seong, P. N., Park, B., & Cho, S. (2014). Evaluation of various
packaging systems on the activity of antioxidant enzyme, and oxidation and color
stabilities in sliced Hanwoo (Korean cattle) beef loin during chill storage. Asian-
Australasian Journal of Animal Sciences, 27(9), 1336–1344.
Kim, Y. H., Huff-Lonergan, E., Sebranek, J. G., & Lonergan, S. M. (2010). High-oxygen
modified atmosphere packaging system induces lipid and myoglobin oxidation and
protein polymerization. Meat Science, 85(4), 759–767.
Kyrana, V. R., Lougovois, V. P., & Valsamis, D. S. (1997). Assessment of shelf-life of
maricultured gilthead sea bream (Sparus aurata) stored in ice. International Journal of
Food Science and Technology, 32, 339–347.
Lan, Y., Shang, Y., Song, Y., & Dong, Q. (2016). Changes in the quality of superchilled
rabbit meat stored at different temperatures. Meat Science, 117, 173–181.
Leygonie, C., Britz, T. J., & Hoffman, L. C. (2012). Meat quality comparison between fresh
and frozen/thawed ostrich M. iliofibularis. Meat Science, 91(3), 364–368.
Lindahl, G. (2011). Colour stability of steaks from large beef cuts aged under vacuum or
high oxygen modified atmosphere. Meat Science, 87, 428–435.
Meat Science 171 (2021) 108292
Liu, D., Li, L., Xia, W., Regenstein, J. M., & Zhou, P. (2013). Biochemical and physical
changes of grass carp (ctenopharyngodon idella) fillets stored at −3 and 0 °C. Food
Chemistry, 140, 105–114.
Łopacka, J., Półtorak, A., & Wierzbicka, A. (2016). Effect of reduction of oxygen con-
centration in modified atmosphere packaging on bovine M. longissimus lumborum
and M. gluteus medius quality traits. Meat Science, 124, 1–8.
Lu, Q., Tang, X., & Dong, J. (2017). A feasibility quantification study of total volatile basic
nitrogen (TVB-N) content in duck meat for freshness evaluation. Food Chemistry, 237,
1179–1185.
O’Keeffe, M., & Hood, D. E. (1982). Biochemical factors influencing metmyoglobin for-
mation in beef from muscles of differing colour stability. Meat Science, 7, 209–228.
Qian, S., Li, X., Wang, H., Sun, Z., Zhang, C., & Guan, W. (2018). Effect of sub-freezing
storage (−6, −9 and −12°C) on quality and shelf life of beef. International Journal of
Food Science and Technology, 53(9), 2129–2140.
Ramanathan, R., Nair, M. N., Hunt, M. C., & Suman, S. P. (2019). Mitochondrial func-
tionality and beef colour: A review of recent research. South African Journal of Animal
Science, 49(1), 9–19.
Renerre, M. (1999). Biochemical basis of fresh meat colour. Proceedings of the 45th in-
ternational conference of meat science and technology (ICoMST), Yokohama, Japan (pp.
344–353). .
Seyfert, M., Hunt, M. C., Mancini, R. A., Hachmeister, K. A., & Unruh, J. A. (2004).
Accelerated chilling and modified atmosphere packaging affect color and color sta-
bility of injection-enhanced beef round muscles. Meat Science, 68(2), 209–219.
Siu, G. M., & Draper, H. H. (1978). A survey of the malonaldehyde content of retail meats
and fish. Journal of Food Science, 43(4), 1147–1149.
Suman, S. P., Mancini, R. A., & Faustman, C. (2006). Lipid-oxidation-induced carbox-
ymyoglobin oxidation. Journal of Agricultural and Food Chemistry, 54(92), 48–53.
Tang, J., Faustman, C., & Hoagland, T. A. (2004). Krzywicki revisited: Equations for
spectrophotometric determination of myoglobin redox forms in aqueous meat ex-
tracts. Journal of Food Science, 69(9), C717–C720.
Van Rooyen, L. A., Allen, P., & O’Connor, D. I. (2017). The application of carbon mon-
oxide in meat packaging needs to be re-evaluated within the EU: An overview. Meat
Science, 132, 179–188.
Wang, F., Holman, B. W. B., Zhang, Y., Luo, X., Mao, Y., & Hopkins, D. L. (2020).
Investigation of colour requirements of frozen beef rolls by Chinese consumers for hot
pot. Meat Science, 162, Article 108038.
Zhang, Y., Qin, L., Mao, Y., Hopkins, D. L., Han, G., Zhu, L., & Luo, X. (2017). Carbon
monoxide packaging shows the same color improvement for dark cutting beef as high
oxygen packaging. Meat Science, 137, 153–159.