Lack of evidence for existence of vitamin D and 25-hydroxyvitamin D sulfates in human breast and cow's milk.

Abstract

The presence of water-soluble vitamin D and 25-OH-D sulfates in human breast and cow's milk was studied. We first confirmed that synthetic vitamin D2 and D3 sulfates could not be hydrolyzed by alkali but by acid. Breast or cow's milk was separated into milk whey containing water-soluble components and milk curd containing crude proteins and lipophilic components. The separated milk whey and curd were hydrolyzed by acid or alkali and each lipid extract was subjected to HPLC analysis. Neither peak due to vitamin D and 25-OH-D was observed in the chromatograms of acid- and alkali-hydrolyzed milk whey, whereas the peaks due to vitamin D3 and 25-OH-D3 were found in the chromatograms of both acid- and alkali-hydrolyzed milk curd and there was no significant difference between the respective peak heights. The eluates corresponding to the respective peaks observed on the latter's chromatograms were collected and subjected to UV, HPLC, GC-MS and GLC to identify the existence of vitamin D3 and 25-OH-D3, respectively. We concluded from these results that neither breast nor cow's milk contained water-soluble vitamin D and 25-OH-D sulfates, whereas they contained fat-soluble vitamin D3 and 25-OH-D3. The concentrations of vitamin D3 and 25-OH-D3 in breast milk were about 125 and 350 ng/liter, while those in cow's milk were about 420 and 270 ng/liter, respectively. The experiments on the transfer of 3H-D3 and 3H-25-OH-D3 perorally dosed to lactating rats into suckling pups through their milk also supported the above conclusion.