Abstract
The amounts of free and bound vitamin B12 in cow's milk were determined microbiologically with the use of the test organism, Lactobacillus leichmannii. Bound vitamin B12 was released by autoclaving the samples in the presence of acid and cyanide. Approximately 95% of the total vitamin B12 in milk was in the bound form. Milk proteins were separated into gross fractions by isoelectric precipitation, coagulation with rennin, centrifugation, and salting-out with ammonium sulfate. Bound forms of the vitamin occurred in all protein fractions, although the whey proteins contained more than the casein proteins, based on the amount of bound vitamin B12 per mg of protein. The amount of bound vitamin B12 in the original milk was accounted for by analysis of the casein and whey protein fractions.
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