AS R E L A T E D TO I M B I B E D WATER

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1 P H Y S I C A L C H A R A C T E R I S T I C S OF MUSCLE T I S S U E S AS R E L A T E D TO I M B I B E D WATER In reviewing the discussions of t h i s Conference r e l a t i n g t o meat hydration and meat tenderness from two years ago it occurred t o me that it would be helpf'ul t o draw together a few of the general aspects of muscle physiology and meat hydration. While t h e discussion which follows probably will not answer any s p e c i f i c questions, it may bring t o light some f a c t o r s which have not been previously discussed in t h i s Confelllence and give a g r e a t e r appreciation of the problems involved. I hope t o be able t o bring i n t o the discussion some of the specific suggestions of t h e Methods Comnittee. In thinking about the o v e r a l l aspects of muscle hydration, it may be w e l l t o review t h e normal e l e c t r o l y t e and water balance i n our s t a r t i n g material the l i v i n g muscle ( I ). Generally speaking, 20 p e r cent of l i v e body weight is composed of e x t r a c e l l u l a r f l u i d s and approximately 50 percent is composed of i n t r a c e l l u l a r f l u i d s. With the exception of the higher - protein content of blood plasma it is very similar t o i n t e r s t i t i a l f l u i d and makes up about 5 per cent of t h e t o t a l e x t r a c e l l u l a r f l u i d. The extrac e l l u l a r water serves as a transport f o r n u t r i e n t and waste materials and is, along with the kidney, l a r g e l y responsible f o r proper balance and constancy i n t h e i n t r a c e l l u l a r f l u i d s. The e l e c t r o l y t e balance and the ionic p a t t e r n of the two f l u i d s is shown i n Figure 1. The striking difference between the makeup of the two f l u i d s lies in t h e i r content of sodium, chloride, potassium and phosphate but it i s a l s o noted that the two f l u i d s have the same t o t a l i o n i c concentrations. While the magnesium and calcium ions are present i n r e l a t i v e l y small amounts they have, as discussed below, a very s i g n i f i c a n t e f f e c t on t i s s u e hydration during post m o r t e m changes. Chloride i s never found i n t r a c e l l u l a r l y and sodium i s probably not a normal c e l l constituent. This c h a r a c t e r i s t i c allows chloride concentrat i o n s t o be used i n estimating i n t e r s t i t i a l f l u i d s and i n d i r e c t l y i n t r a c e l l u l a r f l u i d s. The r a t i o of i n t r a c e l l u l a r t o e x t r a c e l l u l a r water increases during growth but reaches a r e l a t i v e l y constant l e v e l a t maturity if feeding and management remain constant. I n some of our work a t the h r i c a n &at I n s t i t u t e Foundation we have found an increase from 2.0 t o 2.7 i n t h e i n t r a c e l l u l a r - e x t r a c e l l u l a r r a t i o as the rats matured from 3 months t o one year of age. I n the same s e r i e s of experiments we observed a small but s i g n i f i c a n t l y higher I/E r a t i o when rats were f e d corn o i l, than when they were fed lard o i l. These e f f e c t s may be direct o r they may be determined by differences i n h o m n a l b a l a n c e s. This raises the question a8 t o the e f f e c t that hormone supplements f o r livestock may have on the balance between intra- and e x t r a c e l l u l a r f l u i d s. If' this r a t i o could be influenced through feed supplementation it would presumably be desirable t o s h i f t the r a t i o i n favor of higher l e v e l s of i n t r a c e l l u l a r moisture.

2 98 A t t h e time of slaughter m s c l e t i s s u e ph i s normally reduced by t h e formation of l a c t i c acid. Accompanying the drop i n ph i s a decrease i n e l e c t r i c a l resistance and a loss of ATP, IXzring t h i s same period of t i m e the sarccrlermna loses sorue of its semi-penneable c h a r a c t e r i s t i c s and permits a much freer interchange of e l e c t r o l y t e s. The interrelationships between these changes are not e n t i r e l y clear but a few of t h e p r a c t i c a l aspects of these changes have been studied i n 6ome detail. Meat which r e t a i n s a high ph a f t e r s!.alzghter a l s o retains, w h a t has been termed by Callow, as "closed" s t i u c t u r e as contrasted t o an "open" s t r u c t u r e which develops i n meat with an ultimate ph of 5.6 (2). Callow and others have explained t h i s on the basis that because myosin has an i s o e l e c t r i c point of the protein net charge w i l l decrease as it appmaches t h i s ph and t h e meat fibers w i l l be l e s s hydrated. This physical change would then provide f o r f r e e r movement of water and ions through the i n t e r s t i t i a l spaces. That t h i s occurs is a l s o s u g e s t e d by a decreased e l e c t r i c a l resistance as t h e pr drops. This s t r u c t u r a l change would suggest that t h e penetration of pickle would be faster i n meat of low ph. While Bate-Smith (3) observed t h i s t o be t r u e it 'has not been confinned by more recent work of Komenky In t h e l a t t e r work the correlation between rate of penetration and ph (4). was 0.19, According t o Haa3m the post martem drop i n ph explains only onet h i r d of t h e change i n hydration which i s observed and that the breakdown of ATP after death is responsible f o r t h e remaining changes (S), Hamm has indicated t h a t ATP normally binds bivalent ions and upon breakdown of the ATP magnesium and possibly calcium ions become available which can become attached t o the protein making it a cloger knit s t r u c t u r e with less hydrat i o n. Evidence f o r t h i s type of s h i f t can be drawn from the finding t h a t muscle proteins contain less bound magnesium immediately after slaughter than after rigor, F'urther evidence f o r t h i s type of change is found i n t h e patent of Turner and Olson (6). Their patent describes a method f o r preventing the loss of ATP a c t i v i t y and rqyosin s o l u b i l i t y t o r e t a i n t h e water binding properties of meat intended f o r use i n sausage. By freezing meat wlthin three o r four hours of slaughter and defrosting t h e meat i n t h e presence of salt, they have been able t o incorporate meat i n t o an emulsion i n a prerigor state. Under these conditions the soluble protein content remains a t a maximum and the emulsifying and water binding properties are greatest. Ham has reported that it i s possible t o accomplish the same e f f e c t by the addition of salt t o meat i n a pre-rigor state (5). In connection with the post mortem changes i n ph and protein hydration mention m i g h t a l s o be made of the e f f e c t of such changes on the observed c o l o r of rdeat. A number of workers have observed the e f f e c t of physical s t r u c t u r e on the depth of color in meat, The "closed" s t r u c t u r e of meat at a high ph permits reflected light t o pass through a deep l a y e r of pigment and the c o l o r appears t o be a deep dark red. T h i s condition e x i s t s i n dark c u t t i n g beef. If the l i g h t i s s c a t t e r e d by s u p e r f i c i a l layers, as occurs at low ph values, the color w i l l appear much paler even though t h e pigment content is unchanged,

3 99. Because t h i s session i s under the direction of the Research Methods Committee t h i s presentation would be incamplete without some mention of methods f o r measuring water imbibition. The two most popular methods f o r p r a c t i c a l use a= the tube method of Deatherage and aseociates and the HammGrau f i l t e r paper technique. The former method and its application t o water binding studies was described in sane detail by Dr. Deatherage a t our Conference two years ago (7). EiIany of you have used the Hannn-Grau procedure o r some variatlon of it. The study of this method by Wierbicki e t al. ( 8 ) w i l l be of interest t o those unfamiliar with the procedure. The water binding properties of meat as determined by the f i l t e r paper technique have been found t o be closely related t o the ph of meat immediately a f t e r slaughter and d u r i w the development of rigor (9) (8) (5). These investigat i o n s and others (10) (5) show a m l a t i v e l y poor relationship between ph and water binding capacity 24 hours after slaughter o r when t h e ph has reached a constant value. For example, Pedersen (10) found a correlation of only 0.2 between muscle ph and free moisture i n muscles which had an ultimate ph ranging from 5.3 t o 5.6. The discrepancy between the correlations a t the two different times can probably be explained on the basis of the loss of ATP and the loss of s o l u b i l i t y of muscle protein. This would s u a e s t that the correlation between ph and free moisture immediately after slaughter i s t o a considerable extent through the association of decrasing ph with the loss of ATP and myosin s o l u b i l i t y rather than a d i r e c t e f f e c t of ph. This conclusion i s consistent with that of Hamrnts presented above, t h a t ph accounts f o r only one-third of the differences i n muscle hydration. References 1. Gamble, James I, Companionship of water and e l e c t r o l y t e s i n the organization of body f l u i d s. Medical Sciences Vol. 5, Number 1, Stanford University Press, Stanford, California. 2. Callow, E. H, The e l e c t r i c a l resistance of muscular t i s s u e and i t s r e l a t i o n t o curing. Annual Report Food Investigation Board p Bate-Smith, E. C The physiology and chemistry of rigor mortis with special r e f e r a x e t o the aging of beef. Adv. i n Food Research Val, 1: Konnenky, L. and Gy. Gantner On correlations between the ph value of meat and the diffusion of salts. Presented a t Fourth Meeting of European Meat Research Workers, Cambridge, September, Hamm, R Biochemistry of meat hydration. Eleventh Annual Conference on Research, American Meat I n s t i t u t e Foundation, March, Turner, E. W. and F. C. Olson Manufacture of sausage and ground meat products. U. S, Patent No. 2, 874,060. Deatherage, F. E Methods f o r s t u d y i n g t h e water-holding capacity of proteins i n meat. Tenth Annual Reciprocal Meat Conference, p

4 100, 8. Wierbicki, E. and F, E, Deatherage Determination of waterholding capscity of fresh meats. J. Agr, and Food Chem. 6: Briskey, E. J, e t a l The e f f e c t of exhaustive exercise and high sucrose regimen on c e r t a i n chemical and physical pork ham muscle characteristics. J. An. Sci. 18:173-7, 10. Vismer-E edersen, J. 1958, Quality of pork i n r e l a t i o n t o m t e of ph change post mortem. Presented a t Fourth Meeting of European Meat Research Workers, Cambridge, September, 1958 t mm/l OSMOLAR BODY EQUALITY OF THE FLU 1 OS 300 I NTRACELLULAR INTERSTITIAL FLU I O FLUID d 4 I- O I v) z too50 - L K\ Ca. - FIGURE 1 C A T I O N AND A N I O N D ~ S T R I B U T I IN ON INTERSTI T l A L AND INTRACELLULAR GAMBLE (1) FLUIDS.

5 . 101 MR. PEARSON: ThRnk you, George. Dr. Wnsinger b r i e f l y mentioned the effect of ph on muscle protein, and Dr. Wilson f u r t h e r enlareed on t h i s subj e c t, but our next topic deals d i r e c t l y with it, The I s o e l e c t r i c Point of Muscle Protein and Its Relationship t o S o l u b i l i t y and Dispersibility. This paper was o r i g i n a l l y to be given by Dr. Reiner Hamm, but since he could n o t be i n attendance today, we have asked Dr. Deatherage t o take up t h i s subj e c t and present t h e material. Dr. Batherage. #########I

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