THE EFFECT OF METHOD OF SAMPLE PREPARATION UPON THE DETERMINATION OF SOLUBILIZED INTRAMUSCULAR COLLAGEN FROM BOVINE MUSCLE BY QUANTITATION OF ITS HYDROXYPROLINE CONTENT

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Abstract
A total of 91 muscle samples were utilized in a series of three separate but related experiments to evaluate possible effects of method of sample preparation upon the yields of intramuscular hydroxyproline from the heat-soluble, insoluble and combined fractions of bovine muscle tissue. The combined results of these experiments indicate that yields of intramuscular hydroxyproline, and thereby yields of intramuscular collagen, obtained in different studies can not be compared unless the same muscle is evaluated and the same method of sample preparation is employed in both and/or all studies in which yields are to be compared. Results from these experiments may also suggest that sample preparation, in which samples are frozen in liquid nitrogen, results in underestimation of intramuscular hydroxyproline content, and thereby intramuscular collagen content, from the heat-soluble fraction as evidenced by: (1) the substantially lower yields of intramuscular hydroxyproline from the heat-soluble fraction of samples frozen in liquid nitrogen; (2) the fact that added hydroxyproline was completely recovered in the heat-soluble fraction of muscle during preliminary experiments; and (3) the fact that method of sample preparation (freeze drying vs. direct homogenization) did not influence the recovery rate or result in overestimation of added hydroxyproline.