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5 Measurement of Water-Holding Capacity and Juiciness: K.O. Honikel and R. Hamm
5 Measurement of Water-Holding Capacity and Juiciness: K.O. Honikel and R. Hamm
juiciness
K.O. HONIKEL and R. HAMM
5.1 Introduction
A. M. Pearson et al. (eds.), Quality Attributes and their Measurement in Meat, Poultry and Fish Products
© Springer Science+Business Media Dordrecht 1994
126 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUCTS
The structure of the muscle and its substructures, especially the highly
organized insoluble myofibrillar proteins, are responsible for the retention
of (about 75%) water in the muscular tissue. A water:total protein ratio
of 3.3 to 3.6 and a water:myofibrillar protein ratio of about 5 exists in the
lean tissue of the major meat animals, namely, beef and pork.
It is generally accepted that the water in meat is bound in different
ways. A small part of the water within the muscle is the constitutional
water which comprises about 0.5 g H 20 per 100 g protein, i.e. about 0.1 %
of the total tissue water that is located within the protein molecules.
Protein- water binding energies for this water are much greater than those
existing in normal water binding (Fennema, 1977). A further part of tissue
water (5- 10% of total water) shows a relatively restricted mobility
(Hamm, 1972, 1975). According to the classification proposed by
Figure 5.1 Scheme of the various forms of water within a muscle cell. Depending on the pH
and salt concentration, meat protein structures such as myofilaments and fibrils shrink or
swell. On the left (a), the scheme shows the shrunken state where protein chains are close to
each other and a large amount of water is 'free' (horizontal lines). In the swollen state (b), a
considerable part of the 'free' water becomes immobilized (net-like structure). Infinite
swelling leads to dissolving of the protein molecules (i.e. they move among each other
without major interference), which is shown in (c). Around the protein threads (thick lines),
the interfacial water is shown (parallel lines to the protein threads).
128 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUCTS
tissues do not exist and, therefore, it is not clear whether and to what
extent water is ordered in this biosystem.
Water in muscle that is not bound as interfacial water or constitutional
water, is more or less expressible during application of any force but it is
not yet known if the differences in the state of water in muscle have any
meaning in the measurement of WHC by physical methods. Therefore, the
contradictory results of studies on the state of water in muscle mentioned
above are of very limited value for understanding the different changes in
the water-holding properties of meat.
The remarkable changes in WHC occurring during storage and proces-
sing of meat are determined by the extent to which the bulk of the water
is immobilized within the microstructure of the intact or comminuted
tissue (Hamm, 1972). Fennema (1977) called this immobilized bulk water
'entrapped water' because it is physically entrapped in a fashion similar to
that found in gels. The authors prefer the expression immobilized water
(Figure 5.1) because this term has been commonly used since the first
review on the WHC of meat (Hamm, 1960). It should be noted that the
changes or differences in WHC of meat that are of practical importance,
are not related to the fractions of constitutional or interfacial water
(Hamm, 1960, 1972, 1986).
There seems, however, to be a more-or-less continuous transition from
water that is strongly immobilized within the tissue to the 'free water' that
can be squeezed out by very low pressure. It is not possible, therefore, to
present absolute figures for the immobilized part of bulk water because
the amount of immobilized water measured depends on the method used;
nevertheless by using a standardized method one can measure relative dif-
ferences in WHC quite accurately.
-
_?fi.
c:
60 -
50 -
-
m 40 -
Cl
30 -
.c x
Cl
Q) 20 0
3: 10
o
4,0 4,5 5,0 5,5 6,0 6,5 7,0 7,5 8,0
pH of meat
Figure 5.2 Water-holding capacity of beef expressed as an increase in weight of a homo-
genate in dependence of the pH of the meat (Grau and Hamm, 1957). Small cubes of beef
(0.3-0.5 cm diameter) were inserted for several hours in 0.15 M salt solutions at the pH
values indicated. After that time the cubes were gently dried by absorbing tissue and
weighed. The symbols show two sets of experiments with different beef samples.
132 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUCTS
5.5 under normal circumstances due to the formation of lactic acid. The
final pH of meat, however, ranges between 6.8 and 5.4. Furthermore, the
WHC depends on the muscle type (Figure 5.4) and the species of animal
as a result of their varying composition and structure.
15 ..
••••
•
•• PSE
•• •
• • ••
••• x
•
••
x •
•
v. .x. 0
~o
o )( 0 ox 0
~o~x~
OXxxOo 0 0
x
x ~ 0
N
• )( 0
~
OX
<9 x o~ 000
• o 0 0
• 0
.x 0 0
•• • 0
0
OF[)
Figure 5.3 Drip loss of various pork muscles over a wide range of pHI. Three different
muscles were used: (.) longissimus dorsi; (x) semimembranosus muscle; and (0) adductor
muscle. The drip-loss measurements lasted from 24-96 h post-mortem. According to the
released drip within this period of time, limits were set for DFD, normal and PSE pork. If
the time of measurement was shorter, e.g. 24- 48 h post-mortem, the limits of drip loss for
the three classes were reduced to less than I % for DFD, 1-5% for normal and above 5% for
PSE pork.
WATER-HOLDING CAPACITY 133
12 F oSS
10 I
8 1
6~
4~
2L
o --~----~----------------------------------~
o 2 4 6 8 10 12 14 16
days of measurement
Figure 5.4 Drip losses of various bull muscles during 14 days of measurement. Day 0 is
24 h post-mortem. Cubes of meat (25~30 g) as described in the text were used. There was no
sarcomere shortening with all sarcomere lengths being between 1.7 and 1.9 !lm. The pH
values of all four muscles had a mean of 5.55~5.60. The number of samples for each point
was 33. longissimus dorsi, 0; psoas major, +; semitendinosus, A; supraspinatosus, o.
are possible in the same plants under prevailing conditions of the chilling
facilities. Nevertheless, they might not be comparable to those data from
other companies.
0.5 34 52 24 42 58 36 44 59 40 55
4 34 50 16 40 53 31 41 55 34 44 60 51
5 36 53 20 41 56 32 42 56 37 45 57 50
7.5 37 51 28 40 56 29 42 57 35 45 60 49
10 27 49 22 38 55 28 40 55 35 45 55 51
14 33 53 22 39 54 22 41 55 22 44 58 53
17 32 49 21 40 57 36 43 57 41 44 58 54
20 37 48 27 42 58 36 43 58 40 44 60 51
20 35 42 16 39 50 24 41 52 27 43 57 48
23 33 55 31 40 57 32 42 59 33 44 59 54
24 37 47 26 44 58 33 46 59 34 53 59 50
27 37 53 28 39 57 32 40 58 34 42 60 47
30 35 52 26 41 56 32 42 57 34 45 60 56
Means 34.4 50.3 23.6 40.4 55.8 31.0 42.1 56.7 34.3 44.8 58.6 51.5
SD c ±2.8 ±3.4 ±4.6 ±1.6 ±2.3 ±4.3 ±1.7 ±2.0 ±5.2 ±2.7 ±1.6 ±2.7
Out of this wide variety of possible methods, the authors have selected
methods and evaluated them critically and recommend standardized pro-
cedures.
0/0
--. -.
-
--
50 \ •
\ ./'
~ 40 \
I
/
•
•
\
.;
~
§u \
~ 30 \ /
a •\ I
0>
\ • /
.~
c 20 /
\
~
/
0 \ / "$1'
00
L
Ul 10 'r--- u..
u..
«
CD
£
0 4 8 12 16 20 24 28 32 36°C
temperature
Figure 5.5 Influence of the storage temperature between I and 24 h post-mortem on sarco-
mere shortening in beef sternomandibularis muscle; 0% shortening is equal to a sarcomere
length of 1.9 ~m.
WATER-HOLDING CAPACITY 137
(%)
from day 2. oOe
8
7
6
5
Vl
Vl
.S' 4
~3
-0
2
1
0
o 4 8 12 16 20 21.. 28 32
muscle temperature at 0-2 4 hou r s
Figure 5.6 Drip loss of beef muscle cubes (about 30 g) from 1-7 days stored at the first day
post-mortem between O°C and 35°C. Samples were those used in Figure 5.5. The days of
measurement are shown on the right side of the diagram.
5.7.1.1 Method for drip loss. Before carrying out the determination, it is
advisable to record the pH of the sample, the time post-mortem and the
type and age of the animal. Knowledge of these parameters is important
when evaluating the results obtained. The method is described for the
widely used longissimus dorsi muscle.
A slice (2.5 cm thick) of the longissimus dorsi muscle is removed
between the eighth thoracic and first lumbar vertebrae, with only freshly
cut surfaces being used for the measurement. Associated adipose tissue
and parts of spinalis and multifidus dorsi muscles should be removed. The
fascia should remain around the muscle. Room temperature during
cutting should be similar to the temperature of the meat. The slice of
muscle should be weighed and suspended by means of a net or thread
inside a plastic pouch and sealed under atmospheric pressure. The samples
are then held at 0-4 C for at least 24 h. The duration of storage must be
D
reported. The pouches should hang in such a way that the exudate
dripping from the meat does not remain in contact with the meat. At the
end of the measuring period the muscle is taken from the pouch, dried
gently with an absorbing tissue and reweighed. During weighing, care
must be taken that no condensation of water vapour occurs on the cold
surface. Drip loss is expressed as the weight loss in mg.g- 1 of the original
weight of meat or, better, as the percentage of original weight. Finally, the
pH of the muscle should be measured again. If other muscles are used for
drip loss determination, the difference in the way of handling is in regard
138 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUCTS
10
8
0
......, 0
"-
•
6 • '" 0, •
.' .
(f)
"-
(f)
0
4
"-
,
Q. , 0
beef
u
...... pork
2
~
0
Figure 5.7 Relationship between sarcomere length and drip loss in beef and pork muscles.
to the shape of the sample only. As noted above, a cube cut parallel and
perpendicular to the fibre direction is recommended.
Following the drip loss determination, the sample can be used immedi-
ately for cooking loss measurements. If there is a delay before cooking
loss measurement, the sample must be wrapped to avoid drying out of the
surface.
Drip losses b
Days
post-mortem 1 day 3 days 6 days 8 days
3 0.88
6 0.70 0.92
8 0.63 0.87 0.97
14 0.57 0.82 0.93 0.96
unfrozpn
Figure 5.8 Drip loss of thawed beef. The meat was kept unfrozen or frozen post-rigor at the
indicated velocities to about - 18°C. After 2 days of frozen storage, the meat was thawed as
indicated. On reaching one, the cubes were cut and weighed. Measurements started at 2°C.
Figure 5.9 Diagram of results of the FPPM obtained with meat of different WHC: (a) pre-
rigor meat with high WHC; (b) post-rigor meat of higher pH and medium WHC; and (c)
post-rigor meat with low pH and low WHC. The area of the outer ring zone represents the
fluid ring, the centre zone is the area of the meat film, which is spread out by the pressure
applied. (From Hofmann, 1982.)
2
em
Q)
10
--'
U
lfl
:J
8
E
.....
u
6
.....
0 0
c 8 0
Ol
t.
c
L
-
\J 2
:J
--'
0
6.7 65 6.3 6 .1 5.9 5.7 5.5 pH
post rigor
Figure 5.10 Dependency of fluid ring area of various pork muscles (e.g. PSE, normal, DFD)
on the final pH value.
WATER-HOLDING CAPACITY 141
-'" 70 7 -
u
:;
E 60' 6
/' WL
a
~ 50
/
If,
'"E
e
~ 1.0 '"
15 ~
.~:::I 30
fr om R
:;:
'"
OJ
20 /~- ----- --- 1)
OJ /
cr: 10 I 12
/
20 {,O 60 80 100 5 6
Load (kp) Time of pressi ng (m in)
Figure 5.11 Parameters of importance for the filter-paper press method: (a) influence of load
(kp) on the amount of released juice, measured by weight loss (WL) or calculated from the
fluid area (RZ) ( - = meat with lower WHC; - - - = meat with higher WHC); (b) influ-
ence of time of pressing on the area of meat (M) and fluid area (RZ). (See Figure 5.9.)
5.7.2.2 Evaluation of the method. When using exactly 0.3 g of meat, the
WHC can be expressed as the area of the fluid ring (Figure 5.10) or as the
amount of loose water in it and is related either to the weight of sample,
to the total water content, or to the protein content of the sample. With
regard to measurement of WHC for samples of raw, unground tissue from
the muscles, the coefficients of repeatability reported in the literature vary
from 0.68-0.98 (Fewson and Kirsammer, 1960; Gravert, 1962; Fewson et
al., 1964; Brendl and Klein, 1971). The error of variance found by Fewson
et al. (1964) was below 5%. Several modifications of this technique have
been suggested. In most of them the application of defined pressure is
recommended (Wierbicki and Deatherage, 1958) or the amount of
released water is determined by weighing the meat sample or the filter
paper before and after pressing (Hamm, 1960, 1972).
The assumption that almost all of the water released during pressing of
the meat sample is located in the fluid area outside the meat area (Grau
142 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUCTS
Table 5.3 Linear correlation coefficients between high-speed centrifugation losses (30 min
at 50000 xg), capillary volumeter measurements and drip losses and time post-mortema
Drip lossb
Days Centrifugation
post-mortem 1 day 3 days 6 days 8 days 14 day losses at 48h
Centrifugation
loss at 48 h
post-mortem 0.12 0.34 0.46 0.47 0.50
Capillary
volumeter at
48 h post-mortem 0.29 0.37 0.35 0.33 0.30 0.38
% centrifugation loss
Figure 5.12 Influence of the sex of cattle and muscles on centrifugation loss at 48 h post-
mortem. The method used is the high-speed centrifugation method described in the text.
Each group contains 13-34 muscles.
~_-Cylinder guide
Capillary tube
It -I---+---+-__Scale
Test Block_---t.-
~_- Stand
__
Meat sample
Stand base
Figure 5.13 Drawing of the capillary volumeter. The test block contains the gypsum plate.
It can be screwed off and exchanged. The cylinder handle lowers the cylinder (weight about
800 g) onto the meat sample.
mikroliter
_ long.dorsi _ psoasmajor
_ semi tend. _ supraspinam
Figure 5.14 Influence of the sex of cattle and different muscles on capillary volumeter values
measured at 48 h post-mortem. The method is described in the text.
(/)
(/)
6
~
0- 4
....
l:J
0
0-
2
0
50
(/)
J'c __E~~----~R--~~~~~r-~~=-~~--
(/)
0 40
01
C
oX
30 •
0
0
u 20
-ge
10
0 2 3 4 5 6 7 8 days
time of storage
Figure 5.15 Influence of time and temperature of storage on drip loss and cooking loss
of the sternomandibularis muscle of beef. Muscle cubes (about 30 g) were stored at 70°C 0,
SoC 6 and 25°C . within I h post-mortem and held for 24 h at these temperatures. From
day 2 onwards they were stored at O°C . Drip and cooking losses were determined by the
methods described in the text.
(cold shortening) and increases with the time of storage. The cooking
losses of the meat, however, stayed fairly constant within the period of
storage and were independent of the temperature of storage (shortening
on the first day) and the drip released.
Cooking losses depended much more on the end-point temperature
(Figure 5.16) and the speed of heating (Figure 5.17). The higher the final
temperature and the slower the velocity of heating, the higher were the
cooking losses. Between 50°C and 70°C, there was nearly a linear rela-
tionship between the final temperature and cooking losses (Figure 5.16).
These relationships have been investigated in detail by Laroche (1982a,b).
Taking these considerations into account, the authors have worked out a
standardized cooking procedure, which is described for the longissimus
dorsi muscle and has been used for drip loss measurements.
48 -
/x
44 x
40 - //
I 0x' ,6
36 - x .
32 -
/i
I
28 -
//' !,/
x ,0 /
, ,
I •
, l
~ 20 -
.2
I
.° , . .
I.
(J1
.~ 16 I ,
""oo I "
X
v 12 ,,:/ .
I .
8 , I
I /
4 / ~ ./
// /
~ .- .
40 50 60 70 80 90 100°C
temperature
Figure 5.16 Influence of end heating temperature and pH of meat on cooking loss. The
meat was heated at a rate of about 2°C.min- 1 to the indicated end-points. x, pH 5.5; 0, pH
5.8; . , pH 6.3.
75°C) and sealed under moderate vacuum (about 150 mm Hg) to remove
the air trapped between the meat and the wall of the bag. This facilitates
complete immersion of the package in water. If air pockets remain in the
pouch, part of the meat may float above the water level. If the bag is not
sealed, care must be taken that the sides of the bag are in contact with the
meat surface in order to allow optimum heat flow. The mouth of the bag
must remain above the water level. Glass beads or stainless steel rods
should be put in the bottom of the bag in order to keep all of the meat
immersed in the bath.
The pouch is placed in water at 75°C for 50 min and then placed in
running tap water (about 15°C) for 40 min, after which the meat is taken
from the bag, mopped dry and weighed. The heat loss is expressed as
heating loss (in g) per initial weight before cooking (in g) or as percentage
heat loss.
There will be occasions when the history of the meat is unknown or
insufficiently known. In such cases, it is advantageous to measure the
WATER-HOLDING CAPACITY 151
0/0
40
39
38 xxx
I,{)
I,{) 37 xx
.9 ><
><
(Jl
c 36 ><
-'"
0
x
0
u 35
34
0 4 8 12 16 20 24 28 32 36
1/ velocity of heating (sec/oC)
Figure 5.17 Influence of reciprocal heating velocity on cooking losses in pork. The end-
point of heating was 95°C.
moisture content of the meat and relate the loss to total moisture content,
or if appropriate, to dry matter content. The method of moisture determi-
nation is described by Boccard et a/. (1981). Results are expressed as
heating loss (in g)/protein (in g). After measuring heating loss, the sample
may be used for other quality evaluations such as tenderness measure-
ments.
,
I
0/0
16 l- • ••
••
14 l- • • • -
•• •
12 - : I -
• • \ ••
-
\Il
>.
10-
•
0
-0
(T) B • -
•
L..
(l)
13
\Il
6-
•• -
\Il
.2 4- •
•
l
.
0..
L..
-0
• •
2,
:
0 I I
10 20 30 40 50 0/0
Cooking loss after 3 days
Figure 5.18 Comparison of cooking and drip losses of pork longissimus dorsi muscles at 3
days post-mortem. All muscles had an ultimate pH of 5.5-5.75. PSE, cold-shortened and
normal muscles were included in the group. Drip losses were determined between 24 h
and 72 h post-mortem, and cooking losses at 72 h post-mortem according to the method
described in the text but after reaching a final temperature of 95°C.
Meat products are generally prepared by the addition of salt and other
additives. In some products, e.g. cooked or raw ham and bacon, the
muscular structure of the meat remain.s fairly intact. In most of the meat
products, however, the fibrillar structure of the meat is destroyed by
cutting or comminuting. The salt and other additives, water and fatty
tissue are finely comminuted with the meat. In many cases, especially in
cooked sausages, a homogeneous batter is obtained that is stabilized by
heating. During heating, cooking stability (the prevention of water and fat
rendering out) is the most important criterion.
Cooked comminuted meat products of the frankfurter type are the
WATER-HOLDING CAPACITY 153
3500
o /'
/
/'
" 0
o
o
8 ",/ '"
,
~
..
"" 0 . / ./
300 '....... 8 . . . . ,- •
.............. -J:J _ _ - - - 0
~ o
___ 0 ___ , _.
,
••
(g 2500
u
Ul
>
Figure 5.19 Changes in the viscosity of unheated meat batters with increasing salt con-
centrations and without (0) or with (e) 0.75% lecithin as an emulsifier. Viscosity was
measured with a rotational viscosimeter (Haake Rotovisko, Berlin, Germany) as described by
Honikel and Hamm (1983).
WATER-HOLDING CAPACITY 155
~
o
(/)
(/)
52
c
o
-+-'
d
01
:::J
~
'--
-+-'
C
<!J
u
Figure 5_20 Centrifugation losses (45000 xg for 30 min) of unheated meat batters as a
means of determining WHC with regard to salt concentration and lecithin in the batter.
Batters were the same as shown in Figure 5.19. The method is described by Honikel and
Hamm (1983).
28
24
.-.i. 20
• • <III
<J
>.
.~ 12
• • 15
~
0
8 10
- 0 ___
- --0
d
5
Figure 5.21 Influence of salt concentration and lecithin on jelly and fat rendering. The
batter was heated to 95°C within 45 min (open symbols without lecithin; closed symbols
batter with 0.5% lecithin).
WATER-HOLDING CAPACITY 157
WHC is due to three factors: (i) the various forms of water in meat; (ii)
compartmentalization within the cellular and subcellular structures of
meat; and (iii) the changes occurring post-mortem that alter the amount
of water in different forms and compartments.
158 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUCTS
5.11 Summary
References
Bendall, J.R. and Restall, D.J. (1983) The cooking of single myofibers, small myofibre
bundles and muscle strips of beef M. psoas and M. sternomandibularis muscle at varying
heating rates and temperatures. Meat Sci. 8, 93.
160 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUCTS