PMID: 6975800Oct 1, 1981Paper

Protein binding assays for 25-hydroxy, 24,25-dihydroxy and 1,25-dihydroxy metabolites of vitamin D1) in human plasma

Journal of Clinical Chemistry and Clinical Biochemistry. Zeitschrift Für Klinische Chemie Und Klinische Biochemie
E KeckH von Lilienfeld-Toal

Abstract

In this study we describe protein binding assays for 25-hydroxy, 24,25-dihydroxy and 1,25-dihydroxy metabolites of vitamin D in 6 ml of plasma. The procedure involves methanol/methylene chloride extraction of plasma followed by separation and purification of mono- and dihydroxylated vitamin D metabolites on two LH 20 columns with different elution solvents. The separation of dihydroxylated vitamin D metabolites is carried out by high performance liquid chromatography (HPLC). 25-Hydroxy and 24,25-dihydroxy vitamin D are measured by competitive protein binding assays with diluted homogenate from vitamin D-deficient rat kidneys. 1,25-Dihydroxy vitamin D is measured by competitive protein binding assay with diluted cytosol from vitamin D-deficient chick intestine. The concentration of 25-hydroxy vitamin D in human plasma, as determined by this assay, was 12.2 +/- 5.9 microgram/l; of 24,25-dihydroxy vitamin D 1.5 +/- 1.0 microgram/l, and of 1,25-dihydroxy vitamin D 25.8 +/- 11.7 ng/l (means +/- S.D., n = 17, October, Düsseldorf). In April, the concentration of the vitamin D metabolites of the same subjects were: 25-hydroxy vitamin D 3.3 +/- 0.9 microgram/l; 24,25-dihydroxy vitamin D 0.17 +/- 0.04 microgram/l and 1,25-dihydroxy vitam...Continue Reading

Citations

Dec 1, 1993·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·H E van der WielW J van der Vijgh
Sep 1, 1982·European Journal of Pediatrics·E KeckH L Krüskemper
Oct 15, 1984·Clinica Chimica Acta; International Journal of Clinical Chemistry·S ScharlaE Mayer
Dec 1, 1987·Journal of Steroid Biochemistry·C E PorteousH L Makin
May 1, 1993·Annals of the Rheumatic Diseases·H Franck, E Keck
Dec 1, 1987·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·S VukicevićA Stavljenić
Feb 1, 1986·Annals of the Rheumatic Diseases·R M van SoesbergenJ K van der Korst

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