A gas-chromatographic method for the determination of vitamin D in fortified non-fat dried milk

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Abstract
A gas-chromatographic method with temperature programming has been adapted for determining vitamin D in fortified non-fat dried milk by using an acetone-ethanol mixture for protein precipitation and ascorbic acid as an anti-oxidant, followed by extraction with chloroform, with a butylated hydroxyanisole-2,6-di-t-butyl-p-cresol mixture as the second anti-oxidant. Sample purification is accomplished by using multi-bore silicic acid column chromatography with gradient elution, followed by antimony trichloride treatment and 2-stage silicic acid micro-column chromatography. Vitamins D2 and D3 are differentiated and determined by gas-liquid chromatography, with one vitamin as internal standard for the other. An over-all average recovery of 104 per cent. of vitamin D2 was obtained when vitamin D2 was used as the internal standard on samples fortified at levels of 10 to 50 µg per 100 g of non-fat dried milk. Approximate total analysis time was 7 hours.

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