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
5.1 Introduction
The measurement of water-holding capacity (WHC) of meat is carried out

in many different ways all over the world. Some of the methods need only
a few minutes to obtain results, others require days. All of them measure
the inherent ability of the cellular and subcellular structures of meat to
retain excess water compared with the amount of the other muscular con-
stituents. Variability occurs for three reasons:
1. The water molecules are not linked in uniform amounts to the various
cell constituents;
2. These constituents have a variable ability to associate with the water
molecules; and
3. The cellular components are changed by processing.
Various amounts of water are released by the methods applied at different
times after slaughter. In the same piece of meat, the drip loss within 24 h
may amount to 3%, the cooking loss to 25-35% and, with the filter paper
press method, about 60% of the water may be released. From these
results, it is evident that the water must have been lost from different
cellular structures. This means the term water-holding has no uniform
meaning because the methods measure various forms of water retention,
which means that the results may not be comparable with each other.
Thus, everyone who measures WHC must be aware of what is required
to be measured and which method is to be applied for that purpose. In
this respect, the chapter will serve as a guideline.
Juiciness can be looked at as another way of measuring WHC, which is
measured by chewing the meat and using the human senses. While the
physical methods for WHC determination can be described and standar-
dized exactly, in juiciness evaluation even standardization or calibration is
difficult, not to mention the evaluation itself. Calibration of juiciness can
be performed only in a group, where all members of a panel try to agree
on a hedonic scale by judging reference samples. Unlike tenderness or
flavour, which can be calibrated by testing reference samples, this is
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
impossible with juiciness as standardized samples have not been described

or produced.
Furthermore, as already mentioned in chapter 1, juiciness, the trans-
formed sensation of a part of the mouthfeel, is influenced by and influ-
ences the impression of toughness and flavour. Additionally, mouthfeel
has an indistinguishable fat component (chapter 4). Well-marbled beef
appears to be more juicy and is often also more tender than a leaner piece
of meat, even if the water content of the cooked products is equal.
As a consequence, this chapter will not provide sophisticated methods
for the evaluation of juiciness in the same sense as they are presented for
WHC.
5.2 Composition and structure of meat
Meat originates from cross-striated muscular and adhering fatty tissue.

The lean muscular tissue consists of muscle cells arranged in bundles,
which are surrounded by connective tissue. Between the muscle bundles
there is a variable amount of fat (less than 1% and up to 10%), part of
which appears as marbling of the meat. Muscle cells contain 72~ 75%
water, while 21 ~24% of muscle weight consists of different proteins. The
remainder is made up by small amounts of salt, lipids, nitrogen com-
pounds of low molecular weight and traces of carbohydrates. The main
protein groups of meat are the extracellular connective tissue proteins
(mainly collagen) and the cellular sarcoplasmic, myofibrillar, and cytoske-
letal proteins. The connective tissue protein content of muscle accounts
for 0.5~4% of the muscle weight, while the actual muscle cell proteins
account for 15~23% by weight (Linke and Arneth, 1970; Honikel and
Wellhauser, 1993). Even if the connective tissue protein content is fairly
low in comparison with the other muscle cell proteins, a small amount is
quite important for the tenderness and WHC of the meat.
Within the muscle cell, two main groups of proteins are important in
the changes occurring in meat during processing and manufacture. The
first group are the soluble sarcoplasmic proteins, with many of them being
enzymes, which comprise about one third of the total muscle proteins.
The second group of proteins are those that are insoluble at physiological
salt concentration (ca. 0.15 M) and pH (7.0), namely, the insoluble myofi-
brillar proteins. The latter are organized in fibrils and their substructures,
the filaments, and are very important constituents of meat with regard to
various aspects of meat quality.
In cross-striated muscles, from which the meat we eat comes, the fila-
ments and fibrils are subdivided into sarcomeres limited by the Z-discs.
These are the basic contractile units of the muscles. These sarcomeres are
able to contract in the presence of adenosine triphosphate (A TP), which
WATER-HOLDING CAPACITY 127
remains available in the muscles during about the first 10 h post-mortem.

In the pre-rigor state, Ca2 + ions are released into the myofibrillar space
by the membranes surrounding the cell substructures (the sarcoplasmic
reticulum (SR) and mitochondria) causing the muscles to contract (Buege
and Marsh, 1975; Hamm, 1979). Contracted muscles going into rigor
mortis exhibit a change in WHC, which is expressed by an enhanced drip
loss.
5.3 State of water in meat
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
(a) (b) (e)
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).
Fennema (1977), this fraction of tissue water might be defined as inter-

facial water (Figure 5.1). This water is bound at the surface of the
proteins, probably in multilayers and in small crevices. The interactions
result in decreased mobility of the water molecules and represent an inter-
mediate stage between constitutional water and the bulk water with a
reduced vapour pressure and melting point. A part of this water remains
liquid even after freezing below -20 e.
D
The question arises as to whether the remaining bulk of cellular water

(around 90-95%) is free in the physical sense of this term, which means
that it can move without becoming attracted by proteins. The interpreta-
tion of nuclear magnetic resonance (NMR) studies about the state of
water in muscle has aroused some controversy. Numerous authors believe
that essentially all of the water in skeletal muscle exists in a physical state
different from that of pure water or diluted electrolyte solutions (Blan-
shard and Derbyshire, 1975; Peschel and Belouschek, 1979). The alter-
native model, again citing NMR evidence, claims that most of the muscle
water behaves like bulk water in a diluted salt solution but that only a
small fraction of the water adjacent to the protein molecules has a
modified molecular structure. This fraction of tissue water would corre-
spond to the constitutional and interfacial water mentioned above. Some
scientists have concluded from their results that in most of intracellular
water no dramatic changes in the characteristics of molecular motion have
been imposed by the constitutents of the muscle cell (Blanshard and Der-
byshire, 1975; Garlid, 1979). The water within muscle, however, is limited
in motion by the sarcolemma or cell wall.
In addition to the intracellular water, there also exists in muscle an
extracellular space filled with water. Hamm (1986) reported that about
lO% of the total water in living muscle may be associated with the extra-
cellular space. The mobility of the extracellular water is supposed to be
somewhere between that of pure water and that of cellular water. The
dimension of the extracellular space depends on the state of swelling.
Swelling of muscle fibres by increasing pH or addition of divalent cations
decreases the extracellular space, whereas shrinkage of fibres as a result of
development of rigor mortis and fall in pH is associated with an increase
in the extracellular space (Rome, 1967; Offer & Trinick, 1983; Hamm,
1986; Offer and Knight, 1988). Therefore, swelling and shrinkage of
muscle fibres in the intact tissue cause movement of water between the
intracellular and extracellular spaces.
In summary according to the literature it is reasonable to suppose that
water is held in muscle: (i) by capillarity, mostly in the interfilamental
spaces within the myofibrils; (ii) a substantial part is situated in the spaces
between the myofibrils; and (iii) water that is located in the extracellular
space. However, methods for an exact and reliable differentiation between
capillary forces and other forces restricting the mobility of water in animal
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.
5.4 Definition of water-holding capacity

There exists a wide variety of methods in research and practice to measure
WHC (Hamm, 1986). Before we enter into the description and compar-
ison of the methods the authors will try to define the expression WHC
and explain in general the methods used.
WHC is the ability of meat - or more generally of meat systems - to
hold all or part of its own and/or added water. This ability depends on
the method of handling and the state of the system. As the state of meat
and its treatment differ considerably, the meaning of WHC varies widely.
Therefore, the methods applied and the state of meat at the time of mea-
surement must be defined exactly in order to obtain comparable results. In
spite of all the variation in the methods used, there are three main ways of
treatment that can be divided in three different basic procedures for mea-
suring WHC, i.e. (i) applying no force, (ii) applying mechanical force and
(iii) applying thermal force.
5.5 General methodology
5.5.1 Applying no force

To this group of methods belong the measurement of evaporation and
weight loss, free drip, bag drip, weep (Penny, 1977; Honikel et al., 1986),
cube drip and related methods (Howard and Lawrie, 1956), whereby the
meat is left to itself under different environmental conditions. The only
force on the meat is gravity. These methods are very sensitive but time-
consuming, requiring from one to several days and are often hastened by
the other two methods that are discussed below.
5.5.2 Applying external mechanical force

By using positive or negative pressure the WHC of meat can be detected
within a few minutes or up to an hour. In this group are the centrifuga-
tion methods (Wierbicki and Deatherage, 1958; Honikel and Hamm,
1983), the filter paper press method (Grau and Hamm, 1957), the capillary
volumeter method (Fischer et aI., 1976) and imbibing methods (Monin et
al., 1981; Kauffman et al., 1986). With these methods the amount of water
released is higher than with the methods discussed under applying no
force (5.5.1) since the pressure applied forces the release of water from the
intra- and extracellular space of the muscle tissue. In drip loss measure-
ments, only extracellular water exudes from the meat (Offer, 1984). There-
fore, factors must be known to evaluate the actual drip loss of the meat
by these mechanical force methods. The matter becomes even more com-
plicated as the state of meat changes during conditioning and ageing,
which also influences the WHC. Therefore, methods applying mechanical
force reveal, in most cases, only a tendency as to how the meat may
behave during the following days but absolute values that can be recalcu-
lated as drip loss cannot be obtained.
5.5.3 Applying thermal force

Since meat usually is consumed after cooking, the WHC of cooked meat
is of central interest. Cooking losses can be measured by a wide variety of
methods (Bendall and Restall, 1983). During heating, the meat proteins
denature and the cellular structures are disrupted, which have a strong
influence on the WHC of meat. Extra- and intracellular water will be
released by the meat sample on cooking. The influence of the method of
heating and the final temperature have a great effect on WHC. The influ-
encing parameters are, however, not fully recognized. Also the relation-
ships between WHC measurements that apply no force, those applying
mechanical force and those using thermal force are not known.
5.5.4 Choosing a method for measuring WHC

The different methods of measuring WHC in research and practice are
due to the different interest of people who handle meat. For example,
people who slaughter, condition and age meat are mainly interested in
weight and drip losses. They understand WHC in these terms. They apply
methods of no force and external mechanical force for evaluating WHC.
In contrast, those who produce 'ready to eat' meat and also consumers
are interested in cooking losses. They use mainly method (5.5.3) or in
laboratories sometimes in combination with method (5.5.2). Manu-
facturers of meat products are concerned about WHC and the fat emulsi-
fication capacity of salted meat products. They generally use methods
(5.5.2) and (5.5.3)
It becomes evident from the different ways of treatment and interest
that WHC means different things to different people, which explains why
different methods are used to evaluate WHC. Therefore, it is and will be
impossible to find a single method that gives reliable information on all
aspects of WHC that are of practical interest.
5.6 Influence of different factors on WHC

The WHC of all methods used in practice depend on the pH of the meat
(Figures 5.2 and 5.3), which changes after death from around 7.0 to about
70
-
_?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.
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.
5.6.1 Evaporation losses

Evaporation losses depend on the following factors :
1. The surface cover of the muscle tissue, such as adipose tissue, covering
or wrapping material;
2. The size of the sample (large or small);
3. The length of the measuring period;
4. The temperature of meat and chilling room; and
5. The air speed and air humidity within the chiller.
Evaporation losses may be standardized in laboratories but, in practice,
standardization becomes rather difficult. Useful measurements, however,
15 ..
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5.3 5,8 6.3
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.
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.
5.6.2 Drip losses

Drip losses also depend on several other factors namely:
1. size of sample;
2. shape of sample;
3. treatment during the conditioning period (Figures 5.5-5.7) and pro-
cessing (Figure 5.8);
4. temperature of chilling; and
5. length of measuring period (Figure 5.6).
Drip losses in meat samples can be measured under defined conditions
and an optimum procedure can be recommended. The sum of the eva-
poration and drip losses is the total weight loss, in which the individual is
interested.
5.6.3 Filter paper press method ( FP PM)

The FPPM was developed by Grau and Hamm (1957). This method
(scheme shown in Figure 5.9) is easy to apply and very rapid. Thus it has
been widely used. Besides pH (Figure 5.10), the results depend on the fol-
lowing factors:
1. The amount of pressure applied (Figure 5.11);

2. The length of time that the pressure is applied (Figure 5.11); and
3. The 'viscosity' or plasticity of the meat, i.e. the pre-rigor state or after
grinding and salting.
5.6.4 Centrifugation methods for uncooked meat

Besides the factors influencing weight loss, the centrifugation losses
depend additionally on the following conditions:
1. Speed of centrifugation (xg);
2. Length of time of centrifugation; and
3. The plasticity of the meat, which is influenced by state of meat and
additives.
5.6.5 Capillary volumeter method

This method makes use of an apparatus developed by Hofmann (1975),
which is shown and described in Figure 5.13. The measurement is faster
than the FPPM with a single measurement requiring 30-120 s. The results
depend on the duration of the measurement and the amount of pressure
applied.
5.6.6 Imbibing method

The imbibing method of Kauffman et al. (1986) is the procedure devel-
oped most recently. With a 3 s measuring time, this is the fastest available
method. The results depend on the length of imbibing and the time
between cutting the fresh surface and the application of the method.
5.6.7 Cooking losses

In addition to pH (Figure 5.16, Table 5.1) cooking losses depend on:
1. The shape and size of sample;
2. Temperature profile during cooking (Figure 5.17);
3. Final cooking temperature (Figure 5.16); and
4. The environment during cooking (in water on salt solution as shown
in Table 5.1, air, wrapping).
A standardized procedure for cooking concerning shape, size and environ-

ment is possible and will be given. The profiles of heating and final tem-
perature, however, are dependent upon the consumers' attitudes in
different countries and may differ from the standardized method
proposed.
Table 5.1 WHC (cooking loss") as % of muscle, unsalted and salted muscleb homogenate (50% water added) at different post-mortem pH values and
temperatures
Temperature pH 6.8 pH 6.1 pH 5.9 pH 5.5

CC)
Muscle Homogenate Muscle Homogenate Muscle Homogenate Muscle Homogenate
Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted
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
aMeat heated for 30 min to 95°C.

bGround meat (homogenate) includes 50% water, resp. 50%,4% NaCI solution added.
cSD = standard deviation.
Out of this wide variety of possible methods, the authors have selected
methods and evaluated them critically and recommend standardized pro-
cedures.
5.7 Description of methods and evaluation
5.7.1 Drip loss determination

Drip losses can be measured on whole cuts or parts of cuts. For practical
reasons it is advisable to use a piece of meat with a defined shape and
weight. The reason for this is, as already mentioned, that drip loss is
dependent upon the surface area and weight of the sample. In larger
pieces, a smaller percentage of the weight is lost as drip. Furthermore, the
fibre direction in the sample taken for drip loss determination is impor-
tant. Therefore, it is recommended that one generally use a cube of meat
cut parallel to the fibre direction at a weight of about 30-100 g. In special
cases, such as the widely used longissimus dorsi muscle, a slice of a muscle
may be preferred to a cube. Within the European Community (EC), a
recommendation for measuring drip on the longissimus muscle was
proposed 10 years ago by Honikel, as discussed in the following section.
0/0
--. -.
-
--
50 \ •
\ ./'
~ 40 \
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.;
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~ 30 \ /
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00
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Ul 10 'r--- u..
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£
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.
(%)
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
10
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0.7 0.9 1.1 1.3 1.5 1.7 1.9 ( }Jm )

final sarcomere length
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.
5.7.1.2 Evaluation of the method. The method as described above uses a

minimum time requirement of 24 h. A longer period of 2 days or more
lowers the variability and increases the correlation coefficient (Table 5.2)
with regard to long-range storage time. If beef muscle is to be aged for 2
weeks, then the reliability of a 24 h drip-loss measurement (r = 0.57) is
Table 5.2 Linear correlation coefficients between

drip-loss measurements and time post-mortem a
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
an = 565 beef samples .

bDay 0 is 24 h post-mortem.
vplocity of frPe'Zing (-10 bis -7"C) during

12 frppzing thawing
slow slow O,OJOC/min

10
fast slow
slow O,05°C/min fasl 1"C lmin
8
fast 50C/min fast
unfrozpn
5 6 7 days after thawing
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.
rather poor. A 3-day measurement increases the correlation coefficient

considerably to a value 0.82.
The samples should not be taken from the carcass or muscles before the
onset of rigor mortis. Small pre-rigor cubes may shorten and results may
be obtained similar to those shown in Figures 5.5-5.7.
Thawed or slightly frozen samples should be measured with care as
their drip loss depends on the method of freezing and thawing (Figure
5.8). Determining drip for 1 day in this case may give false results.
5.7.2 Determination of water-holding capacity using the jilter paper press

method (FPPM)
The procedure is simple and rapid. It is applicable to both intact and
ground tissue, with and without added water. Heat-denatured meat can
also be used and only 0.3 g of tissue is necessary. The technique requires
only a few minutes and the result is fixed permanently on the filter paper.
There are, however, disadvantages of the FPPM; for example the techni-
que is not applicable to samples containing salt and large amounts of fat
and/or water such as sausage emulsions (Grau and Hamm, 1957; Tsai and
Ockerman, 1981) because their plasticity (fluidity) is very high.
In the original FPPM developed by Grau and Hamm (1957), 300 mg of
muscle tissue are placed on a filter paper (Schleicher and Schull, Dassel,
Germany, 2040b resp., Whatman No. 1 or a similar filter paper) with a
defined moisture content and pressed between two plexiglas plates to a
thin film . The water, which is squeezed out, is absorbed by the filter paper
(Figure 5.9). The area of the ring of fluid, which is obtained by subtract-
ing the area of the meat film from the total area, is proportional to the
140 QUALITY ATTRIBUTES IN MEAT, POULTRY AND FISH PRODUC'I:S
(a) (b) (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.)
amount of water. The pressure produced by tightening the plates with

screws and by hand is so great (see Figure 5.l1) that individual differences
in pressure do not influence the fluid area. This simple technique can be
carried out in a slaughterhouse. The accuracy of the method is satisfac-
tory provided that the sample is weighed precisely.
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.
- I -----,---r - ,----.---- r- - - , - -,----,- ,--

M RZ
(a) (b)
-'" 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.)
5.7.2.1 Method. A piece of filter paper (size about 6 x 6 em) is placed

on a plexiglass plate and 0.2-0.4 g of meat (originally exactly 0.3 g) is
placed in its centre. A second plexig1ass plate is put on top and pressed by
a weight of 50 kg or some other device for 5 min. After removal of the
top plate, the meat area is marked with a pencil and, if necessary, with
light meat (e.g. chicken), also the fluid area. The size of the area is
measured by a planimeter or an electronic area measuring device.
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
and Hamm 1957; Wierbicki and Deatherage, 1958) cannot be correct

because the amount of expressible water, determined by weight loss, is
higher than the amount of water in the fluid area, as shown in Figure 5.11
(Hofmann, 1977; Hofmann et al., 1982). Therefore, the fluid that is
squeezed out by pressing must be adsorbed also by the filter paper below
the meat area. With increasing pressure the amount of released water,
determined by the weight loss of the meat sample, increases to about 60%
of the original weight of the sample until a load of about 40 kg is imposed
(Figure 5.11). A further increase in load does not result in any additional
release of juice. It is interesting that the amount of bound water in the
tissue, which cannot be squeezed out, is in the range of 10-15% of the
original tissue water. The amount of non-expressible water corresponds
closely to the amount of constitutional and interfacial water in muscle (see
section on 'States of water in meat'). The total area is much less depen-
dent on WHC than the fluid area (Figure 5.9). From this observation, it
must be concluded that fluid ring is determined primarily by the meat area
of the meat film rather than by the amount of expressible water
(Hofmann et al., 1982).
The magnitude of force depends on the deformability or plasticity of
the meat sample, which is mainly a function of the protein-protein inter-
actions in the myofibrillar system and, therefore, is correlated with WHC.
In meat with a weak protein-protein interaction (e.g. in the pre-rigor state
at a high pH value), a higher proportion of the total water is immobilized
than in post-rigor meat with a strong protein-protein interaction. Conse-
quently, under a certain pressure, meat with high WHC (high pH) is more
easily deformed than meat with a low WHC (low pH). This explains the
highly significant correlations between FPPM and other methods for the
determination of WHC, which are actually pH correlations. Furthermore,
it must be considered that, under certain circumstances, differences in the
deformability of the meat sample might not be related to differences in
WHC but to other factors. This certainly plays a role if large amounts of
water or fat are added to minced meat. With the FPPM, the deformation
of the meat sample to a thin film occurs very quickly; indeed, after a few
seconds the meat film has reached its maximum size (Figure 5.11). The
loosely bound water is squeezed out at the same rate but the adsorption
of the released fluid by the filter paper takes a longer time. It takes several
minutes for the fluid areas to reach their maximum (Figure 5.11).
Originally the FPPM used the fluid area with exactly 300 mg as a
measure for WHC. Following our proposal, George et al. (1980) ex-
pressed WHC in terms of the ratio of meat to fluid. Hofmann et at. (1982)
proposed using the ratio of meat area to total area because within a
certain weight range (200-400 mg) this ratio is independent of the weight
of sample and determined by WHC only.
In studies on the influence of different factors on WHC of meat, Tsai
and Ockerman (1981) found highly significant correlations (r = 0.88-

0.99) between the FPPM and a centrifugation method. According to
Ubertalle and Mazzocco (1979), the FPPM (Grau and Hamm, 1957) IS
quicker and more easily reproduced than the centrifugation methods.
5.7.3 Determination of water-holding capacity by centrifugation

The centrifugation methods are applicable for intact tissue and ground
meat. In the technique of Bouton et al. (1971, 1972) a weighed intact
muscle sample (3-4 g) is centrifuged at 100 000 xg for 1 h in stainless steel
tubes. Fischer et al. (1976) also found that with centrifugation at
60 000 xg for 30 min good results could be obtained. The juice that is
released from the meat is decanted off as quickly as possible in order to
avoid readsorption of water. The meat sample is removed from the tube
with a forceps, dried with tissue paper and then reweighed to determine
liquid loss. If the residue is dried in the tube at 105°C, the total water
content of the sample can be determined, and WHC can be expressed as
released or bound water as a percentage of total water. The coefficient of
variation is less than 5% and, although the conditions are arbitrary, they
are easily reproducible. The need for a high-speed centrifuge makes it
almost impossible to use this type of method outside a laboratory.
Using a relatively low speed and with ground meat, however, the
remaining amount of juice at the end of centrifugation is very small
(Wierbicki and Deatherage, 1958) because the fluid is readsorbed at the
end of centrifugation. In some techniques, readsorption of water is
avoided by separation of the released juice from the sample. The meat can
be put on a fritted glass disk or glass beads at the bottom of the tube,
thus separating the released juice from the tissue by centrifugation (Wier-
bicki et at., 1957). In the method of Penny (1975), the meat is placed on
perforated teflon disks, covered with pieces of fine nylon mesh and placed
at the bottom of the centrifuge tube, which is partially filled with glass
beads. The juice released by centrifugation can also be adsorbed by using
other absorbing materials like gypsum (Hofman et at., 1980) that is placed
on the bottom of the centrifuge tube.
5.7.3.1 High-speed centrifugation method. In each of two centrifuge

tubes, or a multiple of two, with a volume of about 50 ml, 10 g of meat
are weighed as accurately as possible. After balancing the tubes they are
placed in a fixed angle rotor and centrifuged for 20 min at about 5-10 °C
and about 50 000 xg. In a series of the authors' own unpublished experi-
ments, it was shown that the same rotor and speed must be used. Com-
parison with other rotors and speeds is limited. After the centrifugation,
the tubes are at once turned upside down on filter paper for about a
minute, then the samples are taken by a forceps from the tube, dried
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
an = 480/495 beef samples. bDay 0 is 24 h post-mortem.
gently with absorbent paper and weighed. The centrifugation loss is

expressed as percentage centrifugation loss to the original weight.
5.7.3.2 Low-speed centrifugation method. The procedure is very similar to

the high-speed method discussed above. However, before putting the meat
in the tubes, glass beads or other absorbing materials are accurately
weighed into the tubes, after which the meat is placed above the glass
beads or absorbant. The speed of the fixed angle or swing out rotor
should be from 5000-10 000 xg. After the centrifugation, the tubes should
not be turned upside down but the meat should be removed directly by
forceps and handled as described in the high-speed method.
5.7.3.3 Evaluation of the methods. Different centrifugation methods with

low speeds (1200-4000 r.p.m.) and varying duration (8-20 min) using
meat and tissue homogenates were found to be accurate and reproducible
(Grosse et al., 1979). Highly significant correlations between the results of
different centrifugation methods (about 1000 xg) and the FPPM have
been reported by Brendl and Klein (1971). Other authors, using smaller
samples for the FPPM (0.3-0.5 g) came to the conclusion that the cen-
trifugation method (with about 1000 xg calculated) is more exact and
more reproducible (Gravert, 1962; Steinhauf et ai., 1965). Penny (1975)
found a highly significant correlation. (r = 0.88) between 'spun drip'
(1700 xg) of frozen, thawed meat measured with the centrifugation
method and the amount of 'free drip' (released without centrifugation).
An interesting relationship between high-speed centrifugation losses
(50 000 xg) for 30 min and drip losses is shown in Table 5.3. Drip-loss
measurements for 24-48 h post-mortem samples (day 1) had a rather poor
correlation to the centrifugation losses at 48 h. The correlation coefficients
increased considerably for drip losses for 3-14 days but, in the end, still
% centrifugation loss
young bull steer heifer

sex
_ long.dorsi ::::::J psoasmajor

![§ill semitend. _ supr aspinam
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.
had only a moderate correlation coefficient of 0.50. The causes of this

relatively low relationship are unknown. One reason is easily recognizable
by comparing Figures 5.4 and 5.12. While the drip losses in the four
muscles with which the experiments were carried out are rather different
for supraspinatus muscle only, centrifugation losses are additionally lower
with semitendinosus and psoas major muscles compared with longissimus
dorsi. Further variations between the sexes occur. The reason may be the
plasticity of different muscles, which is important with centrifugation.
5.7.4 Capillary volumeter measurements

A newer type of method for the evaluation of WHC of meat is based on
the application of capillary forces to the muscle tissue (Hofmann, 1975;
Fischer and Hofmann, 1978). A gypsum plate is placed onto the surface
of the intact tissue for a definite time (30-120 s).
The loosely bound water is sucked up into the porous material by the
effect of capillary forces. The air displaced from the capillaries by the
meat juice goes into an V-shaped, calibrated capillary glass tube contain-
ing a coloured fluid (Figure 5.13). The volume of the displaced air read
from the shift of this fluid is equal to the volume of released water and is
inversely proportional to the WHC of the tissue. It is not necessary to
adjust the gypsum to a certain humidity but, for each measurement, a new
disk of gypsum has to be used. The thickness of meat has no influence on
the results. However, the volume of the water sucked up by the porous
material depends on the pressure used. The volume increases with an
~_-Cylinder guide
Capillary tube
It -I---+---+-__Scale
Rising liquid -+-+--__ Cylinder

Cylinder handle
. ~\ Hose connection
Test Block_---t.-
~_- Stand
__
Meat sample
r==~~~~~==~ Base plate
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.
increase in load up to 300 g, then reaches a plateau, while at loads greater

than 1000 g the volume increases again with the rising load. In the low-
pressure range, the capillary forces of the gypsum are probably dominat-
ing. Between 300 g and 1000 g the capillary forces seem to be equal to the
WHC, while at loads greater than 1000 g the meat juice is squeezed out
by the high pressure (Fischer et al., 1976). It could be that the plateau of
the volume-pressure curve indicates the border between extracellular and
intracellular water. Surprisingly, with the capillary volumeter method, the
same values of WHC in terms of volume of released water are obtained if
the tissue is cut either parallel or perpendicular to the fibre direction
(Hofmann, 1975).
5.7.4.1 Procedure for capillary volumeter method. A freshly and evenly

cut surface of lean meat of at least 4 x 4 cm is placed below the gypsum
plate. The gypsum plate is then combined with the capillary and a weight
is placed on the meat for a constant time of 30-120 s. At the end of mea-
surement, the amount (~.tl) of meat juice that penetrates into the gypsum
plate is read on the capillary.
mikroliter
young bull steer heifer

sex
_ 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.
5.7.4.2 Evaluation of the method. The methoJ is highly reproducible, the

coefficient of variation being 4%. It has been used successfully for the
evaluation of pork quality. Fischer et al. (1976) have used simple equip-
ment based on the same principle. A significant correlation coefficient
between the results of the capillary volumeter method and those of the
centrifugation method of Bouton et al. (1972) was found. The correlation
between the capillary volumeter method and the FPPM of Grau and
Hamm (1957) was smaller but was still significant (Fischer et al., 1976).
The correlation between capillary volumeter method and drip losses was
astonishingly poor (Table 5.3). One reason again may be the different
behaviour of various muscles. When comparing Figures 5.4 and 5.14 the
differences become obvious. As already observed in comparison of cen-
trifugation losses (Figure 5.12) and drip losses (Figure 5.4), there are also
differences in WHC between muscles, which are detected with drip loss or
capillary volumeter measurements. It should be mentioned, however, that
capillary volumeter measurements with beef muscles are not easily applied
as the enhanced size of the fibre bundles of beef makes it rather difficult to
get the gypsum plate airtight onto the meat surface.
Nevertheless, the method has its advantages. No weighing of the sample
is required, the data obtained are directly related to WHC and also kinetic
measurements can be carried out. It has the disadvantage that the data
cannot be applied to studies on minced meat or homogenates (e.g. sausage
emulsions).
5.7.5 Imbibing method

The FPPM and the capillary volumeter method are fast and can be
carried out without the need of electric power. The equipment is easy to
carry and can be handled by any trained person. However, Monin et al.
(1981) and Kauffman et al. (1986) indicated the procedure was too com-
plicated. Both groups developed methods which use a simple filter paper,
getting results within seconds.
5.7.5.1 Imbibing method of Kauffman et al. (1986). The method of

Kauffman et al. (1986) uses a muscle surface that is cut and left for 10-
15 min. Then a 45 mm diameter filter paper (Kauffman et al., 1986 used
filter paper No. 300 204 of Schleicher and Schull, Dassel, Germany) is
placed on the meat surface for about 1-2 s. The paper is evaluated imme-
diately on a wetness score of 0-5 (0 = 0% of paper wet, 1 = 20%, 2 =
40%, 3 = 60%, 4 = 80%, 5 = 100% of the paper wet) or the weight
gain is measured at once (weight gain of less than 20 mg to 150 mg).
5.7.5.2 Evaluation of the method. The authors (Kauffman et al., 1986)

found correlation coefficients between drip loss at 48 h and their methods
(either by wetness score or by weight gain) to be 0.90 or higher. More
than 86% of the variation in drip loss could be accounted by variation in
filter paper score. However, pH variation in the meat sample might have
been responsible for the high correlation coefficient.
This method is cheap, easy to handle and, according to the data, very
accurate. There are, however, a few precautions that must be observed.
During the 10-15 min waiting time neither condensation nor evaporation
of water must be allowed to occur. This means that the room temperature
must be very similar to that of the meat and the humidity in the room
must be between 90% and 96%. Second, the method does not work on
meat with a large amount of fat on the surface. Usually this is a minor
problem.
The imbibing method is a rapid alternative to drip loss measurements.
However, its use as a fast method with regard to other WHC measure-
ments must be proven.
5.7.6 Cooking loss measurement

There exists a widespread opinion that meat with a high drip loss also has
a high cooking loss. This, however, has not been shown necessarily to be
the case. In Figure 5.15, an experiment with beef, which was obtained at
1 h post-mortem and stored at various temperatures (O°C, 8°C and 25°C)
within the first day after slaughter is shown. From day 2 on, it was stored
at O°e. Drip loss as already discussed depends on the state of contraction
(/)
(/)
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.
5.7.6.1 Measurement of cooking losses. According to Boccard et al.

(1981) a slice of loin of known weight (70-100 g) and pH is placed in a
thin-walled polyethylene bag (the bag must be waterproof and withstand
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
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.
5.7.6.2 Evaluation of the method. Cooking-loss measurements are used in

a wide range of conditions. As shown in Figures 5.16 and 5.17, many
factors influence cooking losses. Another factor is rendering of the fat in
meat with a high amount of fat. In the literature, fat rendering is often
included in cooking losses. Some scientists, however, like to separate
aqueous cooking juice from liquid fat, especially if this method is applied
to meat batters.
As shown in Figure 5.18, cooking losses and drip losses are apparently
dependent on different meat characteristics. Cooking losses do not depend
on shortening (Figure 5.15) or on the PSE-condition, which can lead to
high drip losses. This is shown on Figure 5.18 where several PSE muscles
and also cold-shortened muscles are compared with normal and unshor-
tened meat samples. Heating causes denaturation of the proteins and dis-
integration of membranes, which has a tremendous influence on the bulk
phase water so that shortening or the amount of extracellular space
becomes unimportant.
,
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.
5.8 Meat products
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
largest in terms of volume and the most thoroughly studied meat

products. In these types of products, WHC is an important characteristic;
thus, the methodology used to detect changes in WHC and the manu-
facturing of cooked sausages will be described briefly.
5.B.1 Manufacturing of cooked sausage

Raw meat of various species (e.g. beef, pork, chicken and others) is cut
in a bowl chopper with the salt, additives, spices and ice or water.
Cutting destroys the cellular and fibrillar structure and facilitates the
action of salt and water, transforming the meat from a state of limited
swelling (Figure 5.la) to one of unlimited swelling (Figures 5.1 b, c) as
explained by Hamm (1975). Salt ions decrease the attractive forces
between adjacent protein molecules, and the added water allows an
increase in volume. The distance between protein molecules is enlarged
and the meat undergoes swelling (Figure 5.1 b). If the swelling of protein
molecules is unlimited, their attractive forces become small enough that
they are solubilized (Figure 5.lc). These salt soluble proteins, discussed
later, are considered to play an important role in meat batter stability.
The degree of swelling and solubilization depends on the pH and rigor
state of the meat, the salt concentration (Hamm, 1972) and the tem-
perature.
Addition of salt not only causes swelling but also results in a change
in the conformational structure of the myofibrillar proteins that in-
creases their hydrophobicity (Voutsinas et at., 1983). The increase in
surface hydrophobicity facilitates the interaction between proteins and
fat particles, forming a covering layer. This allows the hydrophobic
regions of the proteins to interact with each other and to form a
three-dimensional structure upon heating. Thus, the addition of salt
produces three effects: (i) the swelling and dissolving of myofibrillar
proteins, which increases the amount of immobilized water; (ii) the
increase in surface hydrophobicity of muscle proteins that leads to fat
binding; and (iii) the formation of a heat-stable network by hydro-
phobic forces upon heating.
None or little of the water and fat should render out on heating. It is
the intention of the manufacturer to prepare a stable product, which he or
she knows is stable before heating. Therefore, the objective in producing
cooked sausage is to manufacture a stable, cooked sausage. Direct mea-
surements of WHC and fat-rendering of the batter are applied. Also mea-
surements which are supposed to have a relationship to the cooking
stability are used. These include viscosity or emulsification capacity mea-
surements and detection of WHC of the batter before heating, e.g. by cen-
trifugation. Some of the methods used for evaluating cooking stability of
batters will be discussed briefly.
5.8.2 Water-holding capacity of unheated batters

Before cooking, the colloidal batter system is stable due to the low tem-
peratures in the batter and its high viscosity. Gel formation, however, has
not yet taken place. Gel formation on heating depends on the type and
state of the proteins in the batter and the degree of comminution as well
as the heating process itself, i.e. the velocity and final temperature of
heating. Therefore, results obtained with raw material can give hints only
about the stability of the heated product.
The preferred method for evaluating the WHC in the raw batter is the
centrifugation method. Centrifugal forces on the batter are supposed to
release water by differences in density within the batter, but the viscosity
of the batter is also important. The relationship between viscosity and
centrifugation losses is shown in Figures 5.19 and 5.20. In both figures,
the same batters were measured. On increasing the salt content of the
batters above 1%, the authors observed an increase in viscosity and a
decrease in centrifugation losses. This indicated that a higher viscosity,
which is due to a higher interaction between the particles, increased the
3500
o /'
/
/'
" 0
o
o
8 ",/ '"
,
~
..
"" 0 . / ./
300 '....... 8 . . . . ,- •
.............. -J:J _ _ - - - 0
~ o
___ 0 ___ , _.
,
••
(g 2500
u
Ul
>
2000~~__- L_ _~_ _L-~_ _~_ _~_ _~

o 0.5 1.0 1.5 2.0 2.5 3.0 3.5 f..0
% Noel in meat + ice
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).
~
o
(/)
(/)
52
c
o
-+-'
d
01
:::J
~
'--
-+-'
C
<!J
u
% NaCl in mea t + ice
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).
WHC, as measured by centrifugation. In addition to the addition of salt,

lecithin was also added to the batters as an emulsifier in a second set of
experiments. Addition of lecithin decreased the viscosity of the batters
over the entire range of salt concentrations studied. It also lowered cen-
trifugation losses. Thus, salt increased the viscosity and WHC, while
lecithin decreased the viscosity but increased the WHC. Both batter ingre-
dients exhibited different relationships to centrifugation losses and viscos-
ity. However, the opposing relationship between centrifugation "' losses and
viscosity have also been reported in other studies. Pre-rigor, salted meat
has a low viscosity but an excellent WHC due to a low interaction
between actin and myosin (Hamm, 1972). The same situation applies on
addition of di- or polyphosphates to salted meat (Hamm, 1972).
Viscosity is an expression of the interaction between molecules and par-
ticles within an unheated batter. As the examples above indicate, the visc-
osity or interaction between batter components and centrifugation losses
of unheated batters as an expression of WHC may give conflicting results.
5.8.3 Water-holding capacity of heated batters

If viscosity measurements are not always in accordance with centrifuga-
tion losses, how does centrifugation relate to cooking losses? Centrifuga-
tion and heating losses are not always in accordance with each other
(Honikel and Hamm, 1983). Batters with or without lecithin (Figure 5.21)
give different results. Centrifugation losses in batters with lecithin are
always lower than those without lecithin, while cooking losses of batters
with lecithin have higher salt concentrations (above 1.2%) and a higher
cooking loss (cookout) than batters without lecithin. Lecithin, which is an
emulsifier, interacts with hydrophobic regions of the meat proteins
(Honikel and Hamm, 1983), decreasing the ability of the batter to form a
stable gel for coating the hydrophobic surface of the proteins. In unheated
batters, however, lecithin lowers the viscosity by reducing the attractive
forces between protein molecules.
In conclusion, using viscosity and centrifugation measurements as
methods to predict the WHC of heated batters may result in erroneous
28
24
.-.i. 20
• • <III
<J
>.
.~ 12
• • 15
~
0
8 10
- 0 ___
- --0
d
5
1.0 2.0 3.0 4.0

% Noel in meat + ice
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).
predictions of cooking stability. The latter should be measured preferably

by cooking batters under similar conditions, as used for sausage manu-
facturing.
5.8.4 Measurement of water-holding capacity in sausage batters

In principal, the same methods are used as described in the section on
cooking loss measurement in this chapter. Since the rendering of fat will
often occur, the quantity of batters used should be larger than that of the
meat, as weighing of two fluids is difficult. Measuring the volume of the
aqueous phase and the fat phase requires more material. Thus, in actual
tests, a sample of 100-200 g of batter is stuffed in impermeable casings or
in cans and heated to the desired temperature. The batter is then cooled,
preferably overnight and rewarmed to about 45-50°C. (The cans should
not be opened at high temperatures.) Some juice will be lost by over-
pressure. The jelly and fat are fluid and can be poured via a funnel into a
measuring cylinder. After a few moments, jelly and fat will separate and
the volume can be easily measured.
5.8.5 Evaluation of methods

Owing to the changes in the meat proteins described earlier, none of the
methods used for WHC measurements of the raw meat will give a hint
about the WHC of the meat in a cooked sausage batter. If there is any
relationship between the WHC of the meat and batters, the pH of the
meat, which also influences the pH of the batter, will be the decisive
factor. It is easier to measure the pH of the meat than its WHC.
Some papers have reported an intermediate method for WHC between
the meat and batter. The meat is minced or comminuted with salt and
water and heated. The results allow some conclusions but the cookout is
usually somewhat higher than that after comminuting with fat. This can
be explained by the positive action of the fat in batters. The fat particles
stabilize the batter, acting as spacers in the protein network.
In conclusion, the WHC of batters can be measured best in pilot
charges of batters. Predictions by other methods always have their short-
commgs.
5.9 Conclusions about water-holding capacity measurements
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.
WHC can vary widely. As the methods of measurement apply various

forces, different amounts of water are released from the meat. For
example, in drip-loss measurements, a small percentage drip may exude
from the surface within 24 h, while with the FPPM, 60% of the water is
squeezed out within a few minutes. Therefore one should avoid the claim
of measuring the WHC of meat, but report that drip losses or cooking
losses have been measured. The proper method should also be used.
Cooking-loss measurements in slaughter plant are unnecessary but drip-
loss measurements would be the appropriate method. A plant that
produces ready-to-eat meals must cook its meat under conditions similar
to those used to produce its products. In all publications, the methods
must be described carefully.
5.10 Measurement of juiciness
5.10.1 Problems in evaluating juiciness

Evaluation of juiciness can be regarded as another way to evaluate WHC
as stated earlier. Expression of juiciness in objective terms, as with WHC
by applying physical methods, has not been reported to date. Several
obstacles appear to prevent this, including the following.
1: Juiciness is a part of mouthfeel, which is influenced by tenderness and
aroma, so it may be changed by these impressions.
2. Mouthfeel includes the sensation of fat, which enhances the impression
of juiciness.
3. There are no definite units for expressing juiciness, such as ml or per-
centage in WHC.
4. Since there is not an objective measurement, calibration of taste panels
is difficult or impossible.
5.10.2 Evaluation of methods

These problems can be overcome by a series of reference samples with
which the taste panel can be calibrated. However, calibration must be
repeated from time to time as no one can keep reference meat samples for
long. Owing to the many factors influencing mouthfeel, e.g. fat content,
juiciness cannot be related to the water content, neither in the uncooked
nor the cooked meat. If any relationships exist, they are, in most cases,
due to other attributes, such as pH or toughness from cold-shortening.
Thus, evaluation of juiciness by a good, trained taste panel can be
reproducible and result in reduced variability; however, because of the
problems outlined above it will not have the reliability of well-defined
chemical and physical WHC measurements.
Books have been written about sensory evaluation of foods (e.g.

Lawless and Klein, 1991) and hundreds of papers have measured WHC
and evaluated juiciness simultaneously. From these, it becomes clear that
the results are conflicting. The authors recommend that WHC and juici-
ness evaluations be examined separately, since both have some" common
but many different factors that influence them.
5.11 Summary
Water-holding capacity of meat is the ability of meat to hold all or part of

its own and added water. The chemically bound portion of water in meat
is rather small. Fibrillar protein structures, which are the cellular struc-
tures of muscles, and capillary forces retard or prevent the movement of
water molecules. The main water-retaining meat compounds are the myo-
fibrillar proteins, which are organized in myofibres.
The amount of water immobilization depends on the pH value, which
falls from about 7.0 in live muscle to around 5.5 in meat. The minimum
water-holding capacity of meat exists around pH 5.3 since this is near the
isoelectric point of the myofibrillar proteins.
Measurement of water-holding capacity is performed in many ways,
which can be divided into three chief areas: (i) no force is applied other
than the gravity of earth - these methods (e.g. drip-loss measurement)
require a considerable length of time; (ii) mechanical forces such as
pressure or increased gravity speed up the methods described - centrifuga-
tion-loss measurements or the filter paper press method belong to this
group; and (iii) thermal forces release water from the meat upon heating.
The various forces applied cause release of different amounts of water.
A few percentage up to 60% of the weight of meat are released during
measurement. Due to this wide range, comparision of methods is limited.
Therefore, methods should be stated clearly. Also the proper method for
the specific purpose must be used.
Sensory evaluation of juiciness can be regarded as another way to
evaluate water-holding capacity. Even though the results of determining
the water-holding capacity with physical methods can vary among
methods, it is possible to calibrate the methodology and instruments, not-
withstanding the fact that human senses are difficult to calibrate.
References
Bendall, J.R. and Restall, D.J. (1983) The cooking of single myofibers, small myofibre
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5 Measurement of water-holding capacity and

The measurement of water-holding capacity (WHC) of meat is carried out

impossible with juiciness as standardized samples have not been described

5.2 Composition and structure of meat

Meat originates from cross-striated muscular and adhering fatty tissue.

remains available in the muscles during about the first 10 h post-mortem.

5.3 State of water in meat

(a) (b) (e)

Fennema (1977), this fraction of tissue water might be defined as inter-

The question arises as to whether the remaining bulk of cellular water

5.4 Definition of water-holding capacity

5.5 General methodology

5.5.1 Applying no force

5.5.2 Applying external mechanical force

5.5.3 Applying thermal force

5.5.4 Choosing a method for measuring WHC

5.6 Influence of different factors on WHC

5.6.1 Evaporation losses

5.3 5,8 6.3

5.6.2 Drip losses

5.6.3 Filter paper press method ( FP PM)

1. The amount of pressure applied (Figure 5.11);

5.6.4 Centrifugation methods for uncooked meat

5.6.5 Capillary volumeter method

5.6.6 Imbibing method

5.6.7 Cooking losses

A standardized procedure for cooking concerning shape, size and environ-

Temperature pH 6.8 pH 6.1 pH 5.9 pH 5.5

aMeat heated for 30 min to 95°C.

5.7 Description of methods and evaluation

5.7.1 Drip loss determination

0.7 0.9 1.1 1.3 1.5 1.7 1.9 ( }Jm )

5.7.1.2 Evaluation of the method. The method as described above uses a

Table 5.2 Linear correlation coefficients between

an = 565 beef samples .

vplocity of frPe'Zing (-10 bis -7"C) during

slow slow O,OJOC/min

5 6 7 days after thawing

rather poor. A 3-day measurement increases the correlation coefficient

5.7.2 Determination of water-holding capacity using the jilter paper press

(a) (b) (c)

amount of water. The pressure produced by tightening the plates with

- I -----,---r - ,----.---- r- - - , - -,----,- ,--

5.7.2.1 Method. A piece of filter paper (size about 6 x 6 em) is placed

and Hamm 1957; Wierbicki and Deatherage, 1958) cannot be correct

and Ockerman (1981) found highly significant correlations (r = 0.88-

5.7.3 Determination of water-holding capacity by centrifugation

5.7.3.1 High-speed centrifugation method. In each of two centrifuge

an = 480/495 beef samples. bDay 0 is 24 h post-mortem.

gently with absorbent paper and weighed. The centrifugation loss is

5.7.3.2 Low-speed centrifugation method. The procedure is very similar to

5.7.3.3 Evaluation of the methods. Different centrifugation methods with

young bull steer heifer

_ long.dorsi ::::::J psoasmajor

had only a moderate correlation coefficient of 0.50. The causes of this

5.7.4 Capillary volumeter measurements

Rising liquid -+-+--__ Cylinder

r==~~~~~==~ Base plate