The Development of an Improved Milk Substrate for Rennet Coagulation Assay on an Automatic Clot-Timer

A substrate was developed for measuring the milk clotting strength of rennet preparations on an automatic clot-timer. The substrate contained 8 percent pasteurized skim milk solids, 1 percent chloroform, 0.3 percent 200 bloom gelatin, 0.03 M added CaCl2 and was buffered to pH 6.6 with 0.057 M cacodylic acid and 0.042 M triethanolamine. The substrate was shelf-stable for 18 days at room temperatures. It was found that rennet preparations could be standardized to within 1 percent of each other, in terms of milk clotting strength, by use of the substrate on the automatic clot-timer. The method appears to have advantages over conventional rennet standardizing procedures. The research included studies on the effect of chloroform, nonfat dry milk and CaCl2 concentrations; heat and ionizing radiation on the substrate coagulation time