Determination of Vitamin D3 Metabolites Using High-Performance Liquid Chromatography or Immunoaffinity Chromatography†

Our investigation of the analysis of vitamin D3 metabolites has been reviewed. The development of high-performance liquid chromatographic methods for the quantitative determination of 25-hydroxyvitamin D3 3-sulfate and 25-hydroxyvitamin D3, which are the major circulating metabolites of vitamin D3 in human serum/plasma, has been described. The developed methods were applied to the determination of the correlation between the concentration of the sulfate and its genin in healthy subjects and patients with chronic renal failure. The development of immunoaffinity chromatography immobilizing the highly specific anti-1,25-dihydroxyvitamin D3 antibody for the pretreatment of radioreceptor assay of 1,25-dihydroxyvitamin D3, which is the active metabolite of vitamin D3, is also described.

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