Quantitative determination of 25-hydroxyvitamin D3 3-sulphate in human plasma using high performance liquid chromatography.
The quantitative determination of 25-hydroxyvitamin D3 3-sulphate in human plasma was completed using reversed phase high performance liquid chromatography with UV detection (265 nm) and an internal standard method. The vitamin D sulphate fraction was obtained from a plasma specimen with the combined use of a Bond Elut C18 cartridge for solid-phase extraction and a piperidinohydroxypropyl Sephadex LH-20 column for lipophilic ion-exchange chromatography. Separation of the compounds was performed on a YMC-Pack ODS-AM column. The limit of quantitation was 5 ng/mL and the assay was linear from 5 to 50 ng/mL. The proposed method is satisfactory in its accuracy and precision.
Shimada
K
K
Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Mitamura
K
K
Kitama
N
N
D002112:Calcifediol / Q000097:blood*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006801:Humans; D007202:Indicators and Reagents; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D007202:Indicators and Reagents; D002112:Calcifediol