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

doi:10.1007/BF02259302

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 journal › Article › peer-review

4 Scopus citations

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 language	English (US)
Pages (from-to)	141-144
Number of pages	4
Journal	Chromatographia
Volume	13
Issue number	3
DOIs	https://doi.org/10.1007/BF02259302
State	Published - Mar 1980
Externally published	Yes

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

Access to Document

10.1007/BF02259302

Cite this

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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.",

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AU - Clemens, T. L.

AU - Fraher, L. J.

AU - O'Riordan, J. L.H.

AU - Little, C. J.

AU - Dale, A.

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AB - 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.

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