Vitamin D is determined by high performance liquid chromatography (HPLC) in samples containing other fat-soluble vitamins. The vitamin D in the unsaponifiable residue is extracted and separated from interferences by straight phase chromatography, and the fraction corresponding to vitamin D3 is collected and quantitated using the AOAC official final action HPLC method for vitamin D3. Analysis of a synthetic mixture gave reasonable recoveries. The method measures potential vitamin D3 content in milkpowder samples containing 2 IU vitamin D/g in the presence of all known vitamin D3 isomers, vitamin A, and vitamin E.
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