N. F. S. Gault
SUMMARY
INTRODUCTION
The pioneering work of Hamm and his colleagues in the 1950s (see Hamm,
1960) provided the basis of our present knowledge of the physico-
chemical properties of meat which influence its ability to retain water, be
15
Meat Science 0309-1740/85/$03.30 ~" Elsevier Applied Science Publishers Ltd, England,
1985. Printed in Great Britain
16 N. F. S. Gault
The muscles chosen for this study were the M. longissimus lumborum
(LD), M. triceps brachii caput longum (TB) and the M. injhaspinatus (IS).
Thes,e muscles were selected on the basis of their collagen content and
their relative size and shape which facilitated the sampling procedures
used in this work. The LD and TB muscles have a relatively low and
similar collagen content (2'3~o dry weight) while the IS has an
exceptionally high collagen content (13.0 ~o dry weight) with characteris-
tic sheets of connective tissues running throughout its length (Bendall,
1967). Four of each of these muscles were individually taken from the
carcases of twelve young steers of average slaughter weight and age.
Muscles were obtained, when required, three days after slaughter from
the same commercial source. Each muscle was cut transversely to the
muscle fibre direction on a gravity feed slicer into steaks approximately
1.0cm thick, from which discs of meat 3.0cm in diameter (wt 7.5 _+ 1.0g)
were prepared using a cork borer. To facilitate slicing and the preparation
of discs, transverse blocks of muscle and steaks were tempered
indivJLdually at - 2 5 °C until the surfaces were rigid.
Each meat disc was accurately weighed and plgced in a 200 ml capacity
screw-cap polystyrene jar to which 50 ml chilled (~ °C) acetic acid solution
was added. Penetration of acid into the meat was aided by continuous
swirling at 120 rpm in a Gallenkamp Cooled Orbital Incubator for 48 h at
4°C. Untreated meat discs were used as controls. Each treatment was
replicated four times for each individual muscle used.
After equilibration, meat discs were surface dried with paper towelling
and reweighed. Two discs from each treatment were individually cooked
in sealed polythene bags in a water bath at 80°C for 20min and
immediately chilled in an ice-water bath. They were then surface dried as
before and reweighed. The weight of each meat disc, before and after
cooking, was divided by the corresponding raw meat weight to express
change in weight as swelling ratios which served as measures of WHC in
the raw and cooked states.
Cores of meat, 0-9 cm in diameter, were cut from each meat disc parallel
with the muscle fibre direction (where possible) for tenderness assessment
on an Instron Model 1122 Universal Testing Instrument fitted with a
Warner-Bratzler shear device. The average tenderness value for each meat
disc was calculated as the mean of either four (control samples and
those equilibrated in < 0"05M acetic acid) or six (samples equilibrated in
>0"075M acetic acid) measurements of peak shear force.
Chemical analyses
The remaining two meat discs from each treatment were individually
homogenised and made up to 100 ml with distilled water. Measurements
of pH were carried out on these solutions using a digital pH meter fitted
with a combination glass electrode. Titratable acidity was determined on
a 25-ml aliquot of each solution against 001M or 0' 10M NaOH using a
phenolphthalein indicator. Total nitrogen was determined on a 10-ml
aliquot of each solution by the Kjeldahl method (AOAC, 1980) modified
by incorporating a selenium, rather than mercury-based, catalyst. All the
used acetic acid solutions were assessed for total nitrogen (25-ml aliquot),
titratable acidity (5-ml aliquot) and pH by the methods described above.
Titratable acidity values were used to calculate the quantity of acetic
acid (g) in the various equilibrating solutions and meat discs. Values
WHC and tenderness of beef muscles below the ultimate pH 19
obtained from the nitrogen analyses were used to calculate the per cent
total nitrogen content of the raw meat discs leached into the equilibrating
solutions.
Statistical analyses
RESULTS
From preliminary studies carried out with a range of acetic acid solutions
of varying strength (N. F. S. Gault, unpublished data), it was found that
optimum swelling of meat discs had occurred well within a 48-h period
under the conditions described in the 'Materials and Methods' section.
Consequently, this time was considered adequate to ensure that diffusion
of acetic acid into the meat discs had reached a state of equilibrium with
the ex~Lernal solution.
Table 1 shows that there was an expected decrease in the pH of the acid-
treated meat discs, the extent of which was related to the acetic acid
concentration of the equilibrating solutions. However, the results
obtained with the weakest acetic acid solutions were peculiar in that their
corresponding titratable acidity values were less than those of the
untreated controls of normal pH, (Table 1). This was particularly
noticeable with those discs equilibrated in < 0.025M acetic acid solution,
the effect becoming obscure at acetic acid concentrations > 0.05~. These
effects were markedly consistent, not only within each muscle group, but
also between the three muscle types studied, and highlight the complex
nature and uniqueness of each equilibrating treatment.
TABLE 1
Influence o f Acetic Acid C o n c e n t r a t i o n o n its D i s t r i b u t i o n * Between M e a t a n d E q u i l i b r a t i n g S o l u t i o n , a n d o n their p H tO
Characteristics (Acetic Acid C o n c e n t r a t i o n (M) o f F r e s h E q u i l i b r a t i n g S o l u t i o n s ( 8 t o r 165 o b s e r v a t i o n s per m e a n ) )
Musch, Control 0.0025 0.005 0.01 0.025 0.05 0.075 0.10 0.25 (I.50 0.75 1.00 1.25 1.50 2.00 3.00 SEM
{TA ~ 0.03 0.08 0.15 0-23 0.30 0.75 1.50 2.25 3.00 3.75 4.50 6-00 9-00 0.000
:[:TAu 0-06 0.08 0.14 0.20 0.25 0-57 1 . 1 2 1-66 2 . 2 1 2.79 3.36 4.45 6-81 0.035
t T A m 0.06 -- 0.04 0.05 0.07 0.09 fill 0.22 0.39 0.60 0.81 0.97 1.12 1.45 1:97 0.029
%A ~ 0-06 0.15 0-30 0.45 0-60 1 . 5 0 3.00 4.50 6.00 7.50 9.00 12.00 18-00 0.000
LD~
%A -- 0.11 0.17 0.29 0.42 0.55 1.30 2.62 3.89 5.24 6.57 7-89 10.40 15.60 0-055
t%A m 0.81 0.48 0.53 0.67 0.77 0.92 1.62 2-60 3-99 5-11 6.30 7.48 9.82 14.10 0.085
}pH ~ -- 4-41 4.42 3-99 3.90 3.80 3.53 3.29 3.16 3.06 2-97 2.91 2.83 2 - 7 1 0-017
tpH m 5.33 -- - 4.61 4.43 4.31 4.19 4.12 3-84 3.65 3.50 3.44 3.34 3.29 3.22 3.15 0.017
3~TAI -- 0-01 0-02 0.03 0.08 0.15 0-23 0-30 0.75 1,50 2.25 3.00 - 6.00 9.00 0.000
{TA ~ -- 0.04 0.04 0.05 0.09 0-14 0.20 0.25 0.58 1 . 1 3 1.65 2.28 4.50 7-02 0.023
tTA" 0.05 0.01 0.02 0.03 0-05 0.07 0.09 0.12 0.23 0.42 0-62 0.76 -- - 1.50 1.98 0.023
TB { ToA1 - - 0-02 0.03 0.06 0.15 0-30 0.45 0.60 1 . 5 0 3-00 4.50 6.00 12.00 18-00 0.000
++%A" -- 0.08 0.09 0 - 1 1 0.18 0.30 0.43 0.56 1.32 2.64 3.87 5.33 10.58 16-06 0-065
t % A m 067 022 0.27 0.41 0.48 0.58 0.74 0.90 1.59 2.76 3-88 4.95 9.49 13.73 0.080
{pH ~ -- 5.16 4.89 4.66 4.37 4.14 4.01 3 - 9 1 3.60 3.36 3.20 3.10 2.83 2.66 0-012
tpH ~ 5.53 5.21 4.75 4-49 4.36 4.22 4.11 4.00 3.76 3.56 3.45 3.36 3.13 3.02 0.024
{TA 1 001 0.02 0.03 0.08 0.15 0.23 0.30 0.75 1.50 2.25 3-00 6.00 9-00 0-000
{TA u 0.03 0-04 0.05 0.08 0.13 0.19 0.25 0-60 1 . 1 4 1 . 6 6 2.16 4.31 6-55 0.026
t T A " 0.05 0.03 0.03 0.03 0.06 0.08 010 0 - 1 1 0.21 0.42 0.63 0.84 -- - 1.62 2:05 0-038
{ %A t 0"02 0.03 0-06 0-15 0.30 0.45 0.60 1 . 5 0 3-00 4-50 6-00 12.00 18.00 0.000
IS
{ YoA" 0.06 0.07 0.09 0.16 0.29 0.41 0-54 1-35 2.66 3.97 5.22 -- 10.56 15.67 0-027
t %A" 0.57 0.30 0.30 0-36 0.52 0.62 0.78 0.85 1.63 2.78 3.87 4.92 9.22 12-31 0-108
:[:pH~ 5,23 4.95 4.71 4.39 4-14 400 3.92 3.58 3.35 3.20 3.09 2.81 2.67 0.007
t p H m 5.80 5.24 4.88 4-63 4.39 4.26 4.12 4.04 3.77 3.58 3.43 3.32 --- 313 3-02 0-025
* Titratable acidity calculated as acetic acid in all samples, including meat controls.
TA = total content of acetic acid (g): %A = percentage acetic acid present on wt/wt (meat) or wt/vol (solution) basis: Superscriptsy~ u
and rn refer to fresh equilibrating solution, used equilibrating solution and meat, respectively.
WHC and tenderness of beefmuscles below the ultimate pH 21
The loss of nitrogenous material from the meat discs as a function of the
pH of the used equilibrating solutions is illustrated in Fig. 1, where it can
be seen that the greatest loss was found with the IS (30 ~ ) followed by the
TB (27 ~o) and the LD (24 ~). Such losses occurred within the pH range
3.5 to 4.0 which corresponded to original acetic acid concentrations of
0-25~, 0.10M and 0.075r~ for the IS, TB and LD samples, respectively.
There was also a progressive decrease in the loss of nitrogenous material
with decreasing pH to a common value of around 15 ~ for each muscle
type. ]However, at higher pH values, the nitrogen solubility behaviour of
the three muscle types was quite different. While the values for the LD and
TB samples remained between 20 ~o and 25 ~o, there was a progressive
decrease with the IS samples which reached a minimum value of 12 ~o
around pH 5.2.
30
(A)
25
~N
20
15
30
25
~N
20
15
i t t t | | t |
25
~N
2C
15
| t i t n n ,
Fig. 1. Means plots showing the relationship between pH of used equilibrating solution
and per cent nitrogen solubilized from discs of LD (A), TB (B) and IS (C) muscles. (Eight
observations per mean.) SEM (per cent N solubilized) = 0'56 CA), 0.80 (B) and 0'99 (C).
22 N. F.S. Gault
TABLE 2
Regression Analyses of Individual Values Obtained for Cooked Meat Swelling (y) Against
the C o r r e s p o n d i n g Values for Raw Meat Swelling (x) of Discs of LD, T B and IS muscles
(N = 112 per Muscle Type)
(A) ]9,0
1
8. O ~
2.0
7.0 u
.0 6.0
so
4.0
3.0 ~
o
2,0 ~
1,0
, a j ,i
(B) A~A
9.0
2.( 8.0 t
7.0 °~
.0
6.0
5.0
4. o
u') 3.0
2.0 ~
1.0 ~
(c) 9.0
2.( 8.0 t
7.0
o
~ 1.E 6.0
5.0
o
4.0
u'l
3,0 .~
0.E 2.0
1.0 0.
f,-.,-A- , ~ v ' A ' / . , , ,
3.0 3.5 4.0 4.5 5.0 5.5
pH
Fig. 2. Means plots showing the influence of pH of uncooked meat discs on raw meat
swelling (©), cooked meat swelling (O) and cooked meat peak shear force (A) for the LD
(A), TB (B) and IS (C) muscles. (Eight observations per mean for cooked samples, sixteen
observations per mean for raw samples.) SEM (raw meat swelling) = 0.032 (A), 0.025 (B)
and 0.034 (C): SEM (cooked meat swelling) = 0.030 (A), 0.027 (B) and 0.044 (C): SEM
(peak shear force)= 0348 (A), 0.269 (B) and 0-176 (C).
24 N. F. S. Gault
9.0
7.0 \o
JP
6.0
~ y = 106.70e-2"913x y : 22.76e-2.326x
5.0
q.o
3.0
2.0 • ~ e o • •
1.0 _"~ o 0
• ~,i~aiaall.,f~L. _. ~
0.6 1.0 1.4 1.8 2.2 0+6 1.0 1.q 1.8 2.2
Swelling ratio S ~ l l i ng ratio
Fig. 3. The relationships between cooked meat peak shear force and swellingratio in
discs of beef M. longissimus lumborum. (A) raw meat swelling,(B) cooked meat swelling.
peak shear force and swelling ratios of the meat discs from the LD, TB
and IS muscles, respectively. Since all of these Figures indicated an
inverse curvilinear relationship between peak shear force and swelling
ratio, it was considered that the data might best be described by a series of
exponential decay equations of the type:
Y=ae -bx
°
° (a)
tO.O
i.° (A)
9.0
8.0
'S
.~ 7.0 i°
~ G.O
°
*• • Y : 118.0qe_2.997x y : 22.35e - 2 - 3 7 6 x
~ 5.0 o| •
2.0
1.0
O.G 1.0 1.q 1.8 2.2 0.6 1.0 t.~ 1,8 2.2
~OII irlg ratio Sv~l I i ng r a t i o
Fig. 4. The relationships between cooked meat peak shear force and swelling ratio in
discs of beef M. triceps brachii caput longum. (A) raw meat swelling, (B) cooked meat
swelling.
in Figs 3-5. The derived equations and their statistical significance are
summarised in Table 3. The constants a and b for each of the relationships
between peak shear force and swelling ratio were subsequently used to fit
the curves shown in Figs 3-5. Such results confirm the general trends
illustrated in Fig. 2, and show that quantitative relationships, expressed
by exponential decay equations, exist between cooked meat toughness
9.0
r~ 8.0
' Eu
.~ 7.0
~ G .0
,. 5.0
q.0
,•1• Y = 128.38e -3"07fix
°
° \~
-'.~! .
y = 25.71e -2.Gllx
~E 3.0
2.0
1.0
":'~..-
C'.G 1.0 1.q 1.8 2.2 0.6 1.0 1.Zl 1.8 2,2
Svel I i n g r a t i o .~i li ng ratio
Fig. 5. The relationships between cooked meat peak shear force and swelling ratio in
discs of beef M. inJraspinatus. (A) raw meat swelling, (B) cooked meat swelling.
26 N. F. S. Gault
TABLE 3
Regression Analyses of Individual Values of in Peak Shear Force (y) Against the
C o r r e s p o n d i n g Values for Raw Meat Swelling ( R M S ) (x) a n d C o o k e d Meat Swelling
(CMS) (x) of Discs of LD, T B a n d IS Muscles ( N = 112 Per Muscle Type)
and both (a) raw meat swelling ratio and (b) cooked meat swelling ratio,
for the LD, TB and IS muscles used in this study.
DISCUSSION
seen more clearly when the percentage acetic acid contents of the meat
discs and their respective equilibrating solutions are compared. Here, an
apparent distribution balance was reached with the 0.50M (LD) and 0.75M
(TB and IS) acetic acid treatments, beyond which the balance increased in
favour of the equilibrating solutions. Consequently, both the uptake of
acetic acid by the meat discs and their loss of buffering capacity, outlined
above, contributed to the observed pH ranges of the meat discs and their
equilibrating solutions (Table 1).
The nitrogenous material leached from the meat discs (Fig. 1) was most
probably of sarcoplasmic origin, because of the well-established solubility
behaviour of this fraction in contrast to the relative insolubility of
myofibrillar and connective tissue proteins. Lawrie (1979) has also
indicated that non-protein nitrogen and sarcoplasmic protein nitrogen
accou~at for as little as 11 ~o and 26 °/o, respectively of the total nitrogen
content of beef L. dorsi muscle. The maximum losses observed in the
present study could, therefore, account for a substantial proportion of
these components. However, the actual identification of this nitrogenous
material requires further experimental investigation, particularly in the
light of the swelling phenomena discussed later.
The raw meat swelling characteristics of the different muscle types
(Fig. 2) appeared to reflect their differences in connective tissue content,
since the LD and TB samples showed a similar smooth swelling profile
while that of the IS samples was more erratic, exhibiting a pronounced
kink within the pH range 4.3 to 4.0. The swelling behaviour of collagen in
acetic acid solutions is well documented (Gustavson, 1956), and would
account for some of the differences seen over the pH range 4.5 to 4.0
where visible sheets of intramuscular connective tissue in the IS muscle
discs were observed to swell to a greater extent than the myofibrillar
component. Although the diameter of all the meat discs generally
increased to a much greater extent than their thickness, dimensional
changes were too complex to quantify since anisotropic swelling was
regularly observed, particularly with the LD samples, the shape changing
from a circular form to that of an oval.
Despite these differences, the overall raw meat swelling behaviour of
the three muscle types was similar in terms of weight gain. These
relationships (Fig. 2) also bear a striking resemblance to the influence of
acidic pHs on the WHC of ground beef illustrated by H a m m (1960, 1975).
In view of the myofibrillar swelling observed below the IEP by Penny et al.
(1963), it seems reasonable to assume that myofibrillar swelling
28 N. F. S. Gault
meat of pHu > 6 . 0 (Gill & Newton, 1981), only the influence of W H C
below the I E P could be seriously considered for industrial exploitation as
a practical method for tenderizing meat, as the results of the present study
show.
ACKNOWLEDGEMENTS
REFERENCES
AOAC (1980). Official methods of analysis (13th edn.), Association of Official
Analytical Chemists, Washington, DC.
Bendall, J. R. (1967). J. Sci. Fd. Agric., 18, 553.
Bouton, P. E., Harris, P. V. & Shorthose, W. R. (1971). J. Fd. Sci., 36, 435.
Bouton, P. E., Harris, P. V. & Shorthose, W. R. (1972a). J. Fd. Sci., 37, 351.
Bouton, P. E., Harris, P. V. & Shorthose, W. R. (1972b). J. Fd. Sci., 37, 356.
Bouton, P. E., Carroll, F. D., Harris, P. V. & Shorthose, W. R. (1973a). J. Fd.
Sci., 38, 404.
Bouton, P. E., Carroll, F. D., Fisher, A. L., Harris, P. V. & Shorthose, W. R.
(1973b). J. Fd. Sci., 38, 816.
Currie, R. W. & Wolfe, F. H. (1980). Meat Sci., 4, 123.
Dransfield, E. (1977). J. Sci. Fd. Agric., 28, 833.
Fjelkner-Modig, S. & Rud6rus, H. (1983). Meat Sci., 8, 203.
Fredeen, H. T., Martin, A. H. & Weiss, G. M. (1974). J. Fd. Sci., 39, 532.
Gill, G. O. & Newton, K. G. (1981). In: The problems o[dark-cutting in be~[i
(Hood, D. E. & Tarrant, P. V., (Eds)), Martinus Nijhoff, The Hague, p. 305.
Gustavson, K. H. (1956). The chemistry and reactivity of collagen. Academic
Press Inc., New York.
Hamm, R. (1960). Ad~'ances in Food Research, 10, 355.
Hamm, R. (1975). In: Meat(Cole, D. J. A. & Lawrie, R. A. (Eds)), Butterworths,
London, 321.
Khan, A. W. & Lentz, C. P. (1973). J. Fd. Sci., 38, 56.
Lawrie, R. A. (1979). Meat Science (3rd edn.), Pergamon Press, Oxford.
McDougall, D. B., Shaw, B. G., Nute, G. R. & Rhodes, D. N. (1979). J. Sci. Fd.
Agric., 30, 1160.
Miles, C. L. & Lawrie, R. A. (1970). J. Fd. Technol., 5, 325.
Offer, G. & Trinick, J. (1983). Meat Sci., 8, 245.
Penny, I. F., Voyle, C. A. & Lawrie, R. A. (1963). J. Sci. Fd. Agric., 14, 535.