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Meat Science 92 (2012) 7983

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Meat Science
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Meat quality and cooking attributes of thawed pork with different low eld NMR T21
Chunbao Li, Dengyong Liu, Guanghong Zhou , Xinglian Xu, Jun Qi, Peilei Shi, Tianlan Xia
Key Laboratory of Meat Processing and Quality Control, MOE, National Center of Meat Quality and Safety Control, MOST, College of Food Science and Technology,
Nanjing Agricultural University, Nanjing, 210095, PR China

a r t i c l e

i n f o

Article history:
Received 6 April 2011
Received in revised form 28 July 2011
Accepted 2 November 2011
Keywords:
Low-eld NMR
Color
Cooking
Water holding capacity
Pork

a b s t r a c t
A relationship of low eld NMR T2 components to meat quality and cooking attributes of pork was investigated. Longissimus muscle was removed from 23 pig carcasses at 24 h postmortem for meat quality measurements and cooking test. Frozen samples were classied into three groups by LF-NMR T21 of thawed
samples: A (b 40 ms), B (4044 ms) and C (> 44 ms). There were signicant differences (P b 0.05) in pH, lightness (L* value) and pressing loss among the three groups. Cooking time to attain 70 C was slightly lower in
group C than the other groups. Shear force value of cooked samples was not affected by T21. The component
T21 correlated (P b 0.05) with L* value, muscle pH and pressing loss, while L* value correlated (P b 0.05) with
thawing loss and muscle pH. Therefore, combined LF-NMR and color measurements could be a good way to
differentiate water holding capacity of pork.
2011 Elsevier Ltd. All rights reserved.

1. Introduction
In the past 10 years, Bertram and her co-workers have successfully
applied low eld (LF) NMR and other technologies to evaluate pork
quality, especially of water holding capacity (WHC) (Bertram et al.,
2001; Bertram, Purslow, & Andersen, 2002). In LF-NMR analysis,
transverse relaxation components of myowater (T2) within muscle
or meat can be separated into two to three populations representing
the water concentration in different compartments, which are characterized by two to three relaxation times (T2s). Normally, the fastest
relaxation population with a time constant of 010 ms (T2b) is referred to as water tightly associated with macromolecules, and the intermediate population with a time constant of 3050 ms (T21)
represents water located within highly organized protein structures,
e.g. water in tertiary and quaternary protein structures with high
myobrillar protein densities including actin and myosin lament
structures (intra-myobrillar), and nally the slowest population
with a time constant of 100250 ms (T22) represents the water located outside the myobrillar network (extra-myobrillar) (Bertram &
Andersen, 2004). The LF-NMR transverse relaxometry (T2) reects
myowater distribution and mobility and corresponding structural
features in meat which directly affect WHC and drip (Bertram,
Schafer, Rosenvold, & Andersen, 2004; Bertram et al., 2003, 2002;
Pearce, Rosenvold, Andersen, & Hopkins, 2011), and sensory quality
of cooked pork (Bertram, Aaslyng, & Andersen, 2005; Pearce et al.,

2011). All these ndings are extremely important to understand

water distribution and mobility in muscle tissue at different conditions. However, few studies have related LF-NMR data to other quality attributes (e.g., meat color, shear force) and cooking attributes of
meat.
The objective of the present study was to compare meat quality
and cooking attributes among three groups of pork classied by LFNMR T21 of thawed samples.
2. Material and methods
2.1. Sampling
A total of 23 cold pig carcasses (average and standard deviation of
carcass weight were 62.4 kg and 10.8 kg) were randomly selected
from animals, which were reared in the same farm, at 24 h postmortem in a local slaughterhouse. Longissimus dorsi muscle between the
last three lumbar vertebrae was removed from the left side and meat
color of cutting surface was measured. Due to long distance between
the slaughterhouse and the laboratory (approximately 400 km), the
samples for measurements of pH, water holding capacity (thawing
loss, cooking loss and pressing loss), LF-NMR, cooking and shearing
were vacuum packed and frozen at 20 C, and then transported
to the laboratory on dry ice.
2.2. Color measurement

Corresponding author at: Key Laboratory of Meat Processing and Quality Control,
Ministry of Education, College of Food Science and Technology, Nanjing Agricultural
University, 210095 Nanjing, China. Tel.: + 86 25 84395376; fax: + 86 25 84395939.
E-mail address: ghzhou@njau.edu.cn (G. Zhou).
0309-1740/$ see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.meatsci.2011.11.015

After removing from carcasses, cutting surface of samples was

bloomed in the air for approximately 20 min. Five measurements
were performed on each sample. Color parameters were determined

80

C. Li et al. / Meat Science 92 (2012) 7983

using a Minolta colorimeter (CR-300; Minolta Camera Co., Osaka,

Japan) with illuminant D65, a 0 viewing angle and an 8 mm port/
viewing area. Before measurement, the colorimeter was calibrated
with a white tile (mod CR-A43). Color coordinates (L*, a*, b*) were
recorded.
2.3. pH measurement
Muscle pH was measured according to the method of McGeehin,
Sheridan, and Butler (2001). Briey, 1 g of frozen sample was homogenized (Ultra Turrax T25, IKA, Germany) at 6000 rpm for 2 15 s with
a 5 s break in 10 ml of ice-cold buffer containing sodium iodoacetate
(5 mM) and potassium chloride (150 mM), pH 7.0. The pH of the homogenate was recorded with a Hanna 211 pH meter (Hanna, Italy).
2.4. Thawing and cooking
Frozen samples were thawed at +4 C for 16 h. Thawing loss was
calculated as a percentage of weight loss before and after thawing.
Thawed samples were cut into two pieces (one was 2.5 cm thick
and the other was 1.0 cm thick) perpendicular to muscle direction.
The 2.5 cm thick piece was used for cooking test and the 1.0 cm
thick piece was used for pressing test and NMR measurement.
Thawed samples (152.9 15.7 g of weight) were individually
cooked in 80 C water bath till central temperature reached 70 C.
During cooking, central temperature of samples was tracked with a
Testo probe (Pt 100, Testo AG, Germany) connected with a Testo thermometer (Testo 7352, Testo AG, Germany). Temperature recordings
at a 1 s interval were exported and standardized with average weight
and then cooking time to 70 C was calculated. The cooked samples
were chilled to room temperature. Cooking loss was calculated as a
percentage of weight loss before and after cooking.

(T2b, T21 and T22) and their corresponding water population (P2b, P21
and P22) were recorded.
2.8. Statistical analysis
Principle component analysis (PCA) was performed with the program Unscrambler X (version 10.1, CAMO software AS, Oslo, Norway,
2010) to show the differences in the measured variables among all
the samples. All the variables were standardized by multiplying by
1/SD. On scores plot, samples were grouped by variables one by one.
After grouping, the differences in measured variables among
groups were evaluated by one-way analysis of variance. Least squares
means were compared by the Bonferroni's method (Abdi, 2007). In
addition, correlation coefcients between variables were calculated.
The analyses were done with the program SAS 9.12 (SAS Institute
Inc., Cary, NC, USA, 2003).
3. Results
3.1. PCA and sample grouping
Principle component analysis showed that the rst two principle
components (PCs) explained 49% of variation in all measured variables. Scores plot gave a preview on the similarity between samples
(Fig. 1). When each measured variable was individually used for sample grouping, only T21 gave a relatively good separation. Samples in
Group A (average 38.68 ms) were well separated from those in
Groups B (average 42.32 ms) and C (average 44.53 ms) (Fig. 2).
Based on this grouping, other measured variables were further statistically analyzed.
3.2. Carcass weight, pH and meat color

2.5. Shearing
Six to eight 1.27-cm-diameter cylindrical cores were removed
from each cooked sample parallel to the muscle ber direction.
Cores were sheared by a WarnerBratzler shearing machine (Salter
235, Manhattan Kansas, USA) and an average shear force (WBSF)
was calculated for each sample.

Average carcass weight of Group A was much lower (P b 0.05,

Table 1) than those of Groups B and C. Samples in Group C had higher
(P b 0.05, Table 1) pH values than the other two groups. Lightness (L*
value) of muscle samples in Group A was higher (P b 0.05, Table 1)
than the other two groups. There was no signicant difference
(P > 0.05) in a* and b* values between any two groups.

2.6. Pressing test

3.3. Water holding capacity and shearing

Two 2.5 cm in diameter and 1.0 cm thick samples were removed

from each thawed sample using a cylinder sampler (inner diameter,
2.50 cm). Then samples were wrapped with 16 layers of tissue papers
and pressed under a force of 343 N for 5 min using a compression machine (YYW-2, Nanjing Soil Instrument, China). Pressing loss was calculated as a percentage of weight loss before and after compression.

There was no signicant difference in either thawing loss or cooking loss among the three groups (P > 0.05, Table 1). However, pressing loss of thawed samples in Group A was lower (P b 0.05) than
those of the other two groups. No signicant difference existed between Groups B and C (P > 0.05). There was no signicant difference
(P > 0.05, Table 1) in shear force value between any two groups.

2.7. NMR transverse relaxation (T2) measurements

NMR relaxation measurements were performed according to the
method of Bertram, Andersen, and Andersen (2007) with minor modication. Briey, three 3.5 1 1 cm strips were removed along the ber
direction from each thawed sample. The strips were individually
wrapped in paralm membrane, placed a cylindrical glass tube
(18 mm in diameter) and then inserted into the probe of a Niumag
Pulsed NMR analyzer (PQ001, Niumag Corporation, Shanghai, China).
The analyzer was operated at a resonance frequency of 22.4 MHz at
32 C. Transverse relaxation (T2) was measured using the CPMG sequence with a -value of 200 s. Samples were repeatedly scanned for
20 times at a 1 s interval. A total of 3000 echoes were acquired and tted
with a multi-exponential model under the program MultiExp Inv Analysis (Niumag Corporation, Shanghai, China). Three relaxation times

Fig. 1. Principle component analysis (scores plot).On scores plot, samples were grouped
by T21, where A is lower than 40 ms, B is 4044 ms and C is greater than 44 ms.

C. Li et al. / Meat Science 92 (2012) 7983

81

Although samples in Group A seemed to have lower T22 and P21 and
higher P2b, the differences were not statistically signicant (P > 0.05).
3.5. Dynamical change in temperature during cooking
As shown in Fig. 3a, samples in the three groups showed great
variations in temperature prole during cooking. Generally, it
seemed to take slightly longer time for Group A to attain a nal
cooking temperature of 70 C (Fig. 3a and Table 1) compared to
Groups B and C. In addition, temperature prole was dependent
on muscle weight, in which it took longer time for heavier samples
to reach the designated temperature (r = 0.52, P b 0.05, Table 2).
To eliminate weight effect, temperature data were standardized
with average weight by multiplying (actual weight/average weight)
and the temperature proles were shown in Fig. 3b. Groups A (red
lines) and B (blue lines) were generally slower to reach 70 C compared to Group C (black lines). The variation in adjusted cooking
time to reach 70 C among all samples increased (MSE was 4.77)
and the time difference between any two groups increased as well
(Table 1).
3.6. Correlation analysis

Fig. 2. Temperature proles of samples during cooking. (a) Temperature proles

(unadjusted); (b) temperature proles (adjusted). Ai, samples in Group A (i = 1 to
9); Bi, samples in Group B (i = 1 to 7); Ci, samples in Group C (i = 1 to 7). Samples in
each group were arranged in weight ascending order.

4. Discussion

3.4. NMR T2s

As described above, Group A had the lowest T21, with the highest
for Group C and the intermediate for Group B (P b 0.001, Table 1).
Table 1
Comparisons of variables between three groups based on T21 (Means and SEM)1.

Carcass weight (kg)

pH
Color
L*
a*
b*
Water holding capacity
Thawing loss (%)
Cooking loss (%)
Pressing loss (%)
Shear force (kg)
NMR attributes
T2b (ms)
T21 (ms)
T22 (ms)
P2b
P21
P22
Cooking time
(unadjusted, min)
Cooking time
(adjusted, min)

Correlation coefcients among measured variables were listed in

Table 2. NMR T21 showed a positive relationship (P b 0.05) to pressing
loss and pH, but it was negatively (P b 0.05) correlated to color L* and
a*. The P2b correlated negatively (P b 0.05) with adjusted cooking time
(CTadj). Three NMR T2 populations were correlated to each other
(P b 0.05). Lightness (L*) was positively related to thawing loss
(P b 0.05), but negatively (P b 0.05) to pH.

A
(n = 9)

B
(n = 7)

C
(n = 7)

SEM

P value

56.94a
5.75a

70.49b
5.80a

61.29ab
6.05b

9.52
0.19

0.033
0.012

55.40a
6.72
4.46

52.59b
6.29
3.79

52.08b
6.02
3.59

2.53
0.95
0.86

0.032
0.342
0.127

6.82
22.98
36.05a
3.53

4.74
22.54
38.85b
4.10

5.35
23.33
38.48b
3.44

2.32
2.17
2.35
0.75

0.206
0.798
0.052
0.221

2.35
38.68a
186.51
3.40
92.94
3.67
19.42

2.65
42.32b
202.84
3.08
93.11
3.81
19.31

2.24
44.53c
214.85
2.78
94.75
2.47
18.11

0.44
0.97
30.48
0.79
2.79
2.30
3.91

0.216
b 0.001
0.201
0.318
0.402
0.494
0.734

21.04

19.78

17.20

4.77

0.296

P2b, P21 and P22, populations for water closely associated with macromolecules, water
located within the myobrillar protein matrix, and extramyobrillar water, respectively;
a,b,c
Means on the same row with different letters are signicantly different (P b 0.05).
1
T2b, T21, T22, relaxation times for water closely associated with macromolecules,
water located within the myobrillar protein matrix, and extramyobrillar water,
respectively.

4.1. NMR relaxation parameter and water holding capacity of meat

NMR T2 relaxation has been shown to have a good relationship to
WHC of pork, of which T21, T22 and their populations are extremely
indicative for drip of fresh pork and juiciness of cooked product
(Bertram et al., 2003, 2001, 2004). In the present study, T21 of thawed
samples was found to be a relatively good variable to separate pork
samples with different qualities. Group A had relatively low pH and
pressing loss, but higher L* compared to the other two groups.
Group A also appeared to have relatively high thaw loss and long
cooking time (in particular after adjustment with sample weight).
NMR T2 relaxation time showed some relationship to carcass
weight. Group A had lower carcass weight and T21 than the other
two groups. This could be attributed to the difference in animal
growth rate (Pearce et al., 2011). Fast growth rate would increase
myobrillar protein density which would further inuence the

Fig. 3. Typical NMR T2 curves of samples in three groups. Group A, T21 is lower than
40 ms, Group B, T21 is 40 44 ms and Group C, T21 is greater than 44 ms.

82

C. Li et al. / Meat Science 92 (2012) 7983

Table 2
Pearson correlation coefcients between measured variables (N = 23).

L*
a*
b*
TL
CL
PL
pH
T2b
P2b
T21
P21
T22
P22
CT
CTadj

a*

b*

TL

CL

PL

pH

T2b

P2b

T21

P21

T22

P22

CT

CTadj

0.27

0.53
0.70

0.59
0.16
0.29

0.18
0.26
0.04
0.18

0.02
0.08
0.08
0.16
0.40

0.46
0.16
0.41
0.26
0.35
0.26

0.06
0.11
0.10
0.06
0.04
0.10
0.06

0.14
0.06
0.26
0.19
0.33
0.36
0.30
0.16

0.52
0.22
0.42
0.35
0.13
0.51
0.65

0.32
0.05
0.26
0.04
0.31
0.03
0.38
0.24
0.72
0.28

0.06
0.08
0.32
0.35
0.26
0.39
0.35
0.04
0.36
0.40
0.25

0.34
0.03
0.22
0.11
0.26
0.09
0.36
0.35
0.53

0.42
0.10
0.05
0.25
0.14
0.05
0.36
0.03
0.08
0.24
0.12
0.15
0.12

0.32
0.46
0.15
0.28
0.32
0.18
0.15
0.08
0.44
0.25
0.35
0.20
0.27
0.18

0.03
0.38
0.03
0.03
0.16
0.14
0.12
0.02
0.31
0.01
0.17
0.29
0.10
0.52

0.01
0.34

0.22
0.97
0.18

0.72

TL, thawing loss; CL, cooking loss; PL, pressing loss; T2b and P2b, relaxation time and population for water closely associated with macromolecules; T21 and P21, relaxation time and
population for water located within the myobrillar protein matrix; T22 and P22, relaxation time and population for extramyobrillar water; CT, cooking time (unadjusted); CTadj,
adjusted cooking time; W: sample weight before cooking.
P b 0.05.
P b 0.01.
P b 0.001.

intra- and inter-myobrillar myowater characteristics (Pearce et al.,

2011).
Postmortem pH decline is very important for WHC of meat (Bond
& Warner, 2007; Schafer, Rosenvold, Purslow, Andersen, & Henckel,
2002) probably by affecting intramyobrillar and intermyobrillar
water (NMR T21, T22 and their populations) (Bertram, Whittaker,
Andersen, & Karlsson, 2003). Our results also showed a signicant
positive relationship between T21 and pH, i.e., muscle with lower pH
could show more lateral or longitudinal shortening and thus has
less spaces between myobrils with higher potential to reach a proton sink, i.e. higher relaxation time (T21) (Pearce et al., 2011). Relaxation time T21 also correlated positively with pressing loss, indicating
the importance of intramyobrillar water for WHC of meat. As mentioned above, samples with higher T21 would have less intramyobrillar spaces due to shortening and thus show lower water holding
capacity with higher pressing loss (Pearce et al., 2011). In the present
study, the authors did not nd signicant associations between pH
and WHC (thawing loss, cooking loss and pressing loss), which
could be due to alteration of T2s during thawing (Qi et al., 2010). On
the other hand, brightness (L* value) gave a signicant relationship
to T21, pH and thawing loss, indicating that brightness would be an
important indicator for water holding capacity of pork, however van
Laack et al. (1994) showed that pH value at 45 min postmortem
was more important for WHC than L* value.
4.2. NMR relaxation parameter and heat transfer during cooking
During cooking, cooking time (CT) differed slightly among the
three groups and depended greatly on sample weight. When weight
effect was removed, adjusted cooking time (CTadj) differed greatly
between groups and it correlated negatively with P2b, i.e., a lower
water population associated with macromolecules resulted in an increased cooking time, which indicates that tightly bound water in
muscle could play an important role in heat transfer during cooking.
In fact, water content in muscle tissue may determine the thermal
conductivity during cooking (Perez & Calvelo, 1984), if more free
water in the extramyobrillar space in the sarcoplasm (intermyobrillar), between muscle bers (interfascicular) and between the
muscle fasciculi (extrafascicular) and immobilized water (intramyobrillar) were available (Pearce et al., 2011), heat transfer
from surface to center would be faster. Conversely, heat transfer
would be slow if protein-associated water (charged hydrophilic
groups on the muscle proteins tightly bound water) (Pearce et al.,

2011) is high. In the present study, thawed samples in Group C had

slightly higher content of intramyobrillar water (P21), and most
importantly, intramyobrillar and extramyobrillar water in these
samples was more mobile (higher T21 and T22), which would be benecial for heat transfer during cooking. Meanwhile, structural
changes in protein (intramolecular antiparallel (beta)-sheets and
(alpha)-helical structure) during cooking may be associated with
simultaneous alterations in water distribution in muscle (P2b, P21)
and this process would depend on pH value of samples (Bertram,
Kohler, Bocker, Ofstad, & Andersen, 2006).
Although there was no signicant difference in cooking loss among
the three groups, some changes in water distribution and mobility
could have taken place. Tornberg (2005) attributed cooking loss of
meat to the denaturation of sarcoplasmic proteins between 40 and
60 C, transverse shrinkage of the ber between 40 and 60 C and longitudinal shrinkage of connective tissue network and muscle bers between 60 and 70 C. The cooking-induced shrinkage of the myobrils
was mainly induced by denaturation of myosin rods and light chains between 53 and 58 C (Bertram, Wu, van den Berg, & Andersen, 2006).
5. Conclusions
In the present study, NMR T21 of thawed pork was shown feasible
to classify pork with different qualities. Samples were classied by LFNMR T21 of thawed pork into three groups that showed signicantly
different pH, L* value and pressing loss. The three groups also showed
slightly different cooking time to attain 70 C. Our results conrmed
that it could be feasible to differentiate water holding capacity of
pork by a combination of LF-NMR measurement and color evaluation.
Acknowledgments
This work was funded by 30901126, 200903012, CARS-36-11 and
the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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