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In previous studies of the folate content of human milk, samples were prepared for assay by a method that resulted in a turbid solution that was then assayed by a turbidimetric microbiological method. We have used an improved microbiological assay in which the milks were treated with rennin to precipitate casein and heated in a buffered ascorbate to coagulate lactalbumin and lactoglobulin. Milks were obtained serially from nursing mothers for periods ranging from 1 day to 6 months postpartum. The results showed that the folate in human milk has few glutamate residues since treatment with a purified folate conjugase preparation release no additional folate activity for Lactobacillus casei. Colostrum is relatively low in folate, but milk folate increases as lactation proceeds. During each stage of lactation there was great variation in milk folate content among the women. In the case of a folate-deficient woman, supplementation with folic acid resulted in a prompt increase in milk folate level.
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