Determination of 25-hydroxyvitamin D₃ in human plasma using HPLC with UV detection based on SPE sample preparation

Interest in the metabolism and physiological action of vitamin D is increased exponentially. The most important metabolites of vitamin D are 25-hydroxyvitamin and 1,25-dihydroxyvitamin D₃. The aim of the study was to develop a rapid and simple HPLC method for the measurement of 25-hydroxyvitamin D₃...

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Published in:Journal of separation science 2009-09, Vol.32 (17), p.2953-2957
Main Authors: Kand'ar, Roman, Zakova, Pavla
Format: Article
Language:English
Subjects:
25-hydroxyvitamin D3
Adult
Analysis
Biological and medical sciences
Calcifediol - blood
Chromatography, High Pressure Liquid - instrumentation
Chromatography, High Pressure Liquid - methods
Female
General pharmacology
HPLC with ultraviolet detection
Humans
Male
Medical sciences
Pharmacology. Drug treatments
Reference Values
Retinyl acetate and solid-phase extraction
Sensitivity and Specificity
Solid Phase Extraction - methods
Ultraviolet Rays
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Zakova, Pavla
description Interest in the metabolism and physiological action of vitamin D is increased exponentially. The most important metabolites of vitamin D are 25-hydroxyvitamin and 1,25-dihydroxyvitamin D₃. The aim of the study was to develop a rapid and simple HPLC method for the measurement of 25-hydroxyvitamin D₃ in human plasma. A method for the measurement of 25-hydroxyvitamin D₃ using HPLC with UV detection and investigation into the extraction techniques with regard to stability and recovery are described. For the separation, RP column LiChroCart 125-4, Purospher RP-18e, 5 μm, was used. The mixture of methanol and deionized water (95:5 v/v) was used as mobile phase. The analytical performance of this method is satisfactory: the intra- and inter-assay coefficients of variation were below 10%. Quantitative recoveries from spiked plasma samples were between 92.0-103.2%. The LOD was 10 nmol/L. The preliminary reference range of 25-hydroxyvitamin D₃ in a group of blood donors is 62 ± 26 nmol/L.
doi_str_mv 10.1002/jssc.200900213
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subjects 25-hydroxyvitamin D3
Adult
Analysis
Biological and medical sciences
Calcifediol - blood
Chromatography, High Pressure Liquid - instrumentation
Chromatography, High Pressure Liquid - methods
Female
General pharmacology
HPLC with ultraviolet detection
Humans
Male
Medical sciences
Pharmacology. Drug treatments
Reference Values
Retinyl acetate and solid-phase extraction
Sensitivity and Specificity
Solid Phase Extraction - methods
Ultraviolet Rays
title Determination of 25-hydroxyvitamin D₃ in human plasma using HPLC with UV detection based on SPE sample preparation
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