PMID: 6974771Jan 1, 1981Paper

A method for simultaneous determination of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in human plasma by using two steps of high-performance liquid chromatography

Journal of Nutritional Science and Vitaminology
T OkanoT Matsuo

Abstract

A method for simultaneous determination of 25-hydroxyvitamin D2 (25-OH-D2) and 25-hydroxyvitamin D3 (25-OH-D3) in human plasma has been developed by using two steps of high-performance liquid chromatography (HPLC). Lipids extracted from 0.51 ml of human plasma were first subjected to the preparative HPLC using a Nucleosil 5C18 column (reversed-phase type) and a 25-OH-D fraction containing 25-OH-D2 and 25-OH-D3 was separated. The separated fraction was subsequently subjected to the analytical HPLC using a Zorbax SIL column (straight-phase type). Since the peaks corresponding to 25-OH-D2 and 25-OH-D3 were clearly separated from one another on the chromatogram of the analytical HPLC, the metabolites could be simultaneously determined by estimating the respective peak heights. When the fractions corresponding to the respective peaks were separately collected by repeatedly applying rather large quantities of human plasma and were subjected to gas chromatography-mass spectrometry (GC-MS), they were identified as containing 25-OH-D2 and 25-OH-D3, respectively. The proposed method was applied to plasma samples of human adults taking 400 I.U./day of vitamin D2 for 8 weeks and the values were 22.5 +/- 8.1 ng/ml for 25-OH-D3 and 11.5 +/- ...Continue Reading

Citations

Jul 1, 2000·Environmental Health and Preventive Medicine·K NakamuraM Yamamoto
Apr 4, 2003·Aging Clinical and Experimental Research·Kazutoshi NakamuraMasaharu Yamamoto
Mar 28, 1983·Clinica Chimica Acta; International Journal of Clinical Chemistry·D J Trafford, H L Makin
Dec 1, 1987·Journal of Steroid Biochemistry·C E PorteousH L Makin
Jan 1, 1990·Journal of Steroid Biochemistry·V K Agarwal
Jun 1, 1997·Journal of Pharmaceutical and Biomedical Analysis·S MasudaT Kobayashi
Mar 7, 2000·Journal of Pharmaceutical and Biomedical Analysis·M D Luque de CastroJ M Quesada
Dec 1, 1994·Acta Paediatrica Japonica; Overseas Edition·H ShimizuT Kobayashi
Jan 2, 2007·The Journal of Steroid Biochemistry and Molecular Biology·Bruce W Hollis, Ronald L Horst
Jun 1, 1984·Scandinavian Journal of Clinical and Laboratory Investigation·I HolmbergM Sturén
May 9, 2000·The American Journal of Clinical Nutrition·K NakamuraM Yamamoto
Nov 11, 2005·The American Journal of Clinical Nutrition·Kazutoshi NakamuraMasaharu Yamamoto
Jul 29, 1988·Journal of Chromatography·A P De LeenheerR M Bauwens

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