Biological generation of tritiated vitamin D3 metabolites and their purification by high-performance liquid chromatography

T. L. Clemens, L. J. Fraher, J. L.H. O'Riordan, C. J. Little, A. Dale

Research output: Contribution to journalArticle

Abstract

A simple, reproducible method for the biological synthesis of tritiated 24,25-dihydroxycholecalciferol, 25,26-dihydroxycholecalciferol and 1,25-dihydroxycholecalciferol is described. Kidney homogenates from both vitamin D deficient and replete chicks were used in vitro to generate these dihydroxylated metabolites using 25 (23,24-3H) hydroxycholecalciferol as the substrate. Tritiated products were purified by Sephadex LH 20 chromatography followed by high-performance liquid chromatography; the identity of each metabolite was established by chromatography with authentic crystalline preparations.

Original languageEnglish (US)
Pages (from-to)141-144
Number of pages4
JournalChromatographia
Volume13
Issue number3
DOIs
StatePublished - Mar 1980
Externally publishedYes

Keywords

  • High-performance liquid chromatography
  • Renal hydroxylase enzymes
  • Sephadex LH 20
  • Tritiated dihydroxycholecalciferol metabolites

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Organic Chemistry

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