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Occurrence of Vitamin D Sulfate in Human Milk Whey¹

Endocrinology and Mineral Metabolism, VA Medical Center and School of Medicine, Case Western Reserve University, Cleveland, OH 44106 and ^* Department of Nutrition, University of Guelph, Guelph, Ontario, Canada, N1G 2W1

Following reports that vitamin D sulfate is the major source of vitamin D activity in human milk, we investigated the presence of this compound in milk whey using a modification of techniques for the determination of vitamin D metabolites in plasma. Synthetic cholecalciferol sulfate, ergocalciferol sulfate and [³H]cholecalciferol sulfate were prepared by reacting radioactive cholecalciferol or nonradioactive cholecalciferol or ergocalciferol with sulfamic acid in pyridine. The products were purified sequentially by Sephadex LH-20 and high pressure liquid chromatography. The purified products were chromatographically homogenous, exhibited an ultraviolet absorption spectrum identical to that of standard cholecalciferol, demonstrated a sulfonate ester linkage and upon saponification yielded the parent vitamin. Milk whey was extracted with methanol:methylene chloride (1:2, v/v) using [³H]cholecalciferol sulfate to estimate recovery of the compound. The extract was purified by chromatography on silica cartridges and reverse phase high pressure liquid chromatography and was quantitated by ultraviolet absorption (UV). Although added cholecalciferol sulfate was readily detected in human milk whey samples, no endogenous vitamin D sulfate was found (detection limit 1 ng/ml). The results indicate that vitamin D sulfate is not a major source of vitamin D activity in human milk.

KEY WORDS: • milk whey • vitamin D sulfate

¹ Supported by the Veterans Administration, the National Institutes of Health, the National Science and Engineering Research Council of Canada and the Ontario Ministry of Agriculture and Food.

² To whom reprint requests should be sent: VA Medical Center, Cleveland, OH 44106.

Manuscript received 4 April 1980.

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