Quantitation of the main metabolites of vitamin D in a single serum sample. I. Extraction, separation and purification of metabolites

Clinica Chimica Acta; International Journal of Clinical Chemistry
L Aksnes

Abstract

A method for extraction, separation and purification of the main serum metabolites of vitamin D from a single serum sample is described. The method involved extraction of serum by diethylether and separation and purification of vitamin D, 25-OHD and the dihydroxymetabolites 24,25-(OH)2D, 25,26-(OH),2D and 1,25-(OH)2D by elution in three steps from a short open silicic acid column. The eluted vitamin D metabolites were further separated and purified by high pressure liquid chromatography (HPLC). The HPLC systems described separated the D2 and D3 forms of vitamin D, 25-OHD, 1,25-(OH)2D, and probably also 24,25-(OH)2D and 25,26-(OH)2D. The metabolites were purified by the methods described for further quantitation by UV-absorption or competitive protein binding assays, and were found to be homogenous on re-chromatography with different HPLC systems. Good recoveries were obtained for all the metabolites.

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Citations

Dec 1, 1989·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·E DahlJ Halse
Dec 1, 1994·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·T YamateY Seino
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Jul 29, 1988·Journal of Chromatography·A P De LeenheerR M Bauwens

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