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Quantitative determination of 25‐hydroxyvitamin D 3 3‐sulphate in human plasma using high performance liquid chromatography
Author(s) -
Shimada Kazutake,
Mitamura Kuniko,
Kitama Nami
Publication year - 1995
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130090508
Subject(s) - chromatography , chemistry , cartridge , high performance liquid chromatography , detection limit , sephadex , extraction (chemistry) , solid phase extraction , human plasma , mechanical engineering , biochemistry , engineering , enzyme
The quantitative determination of 25‐hydroxyvitamin D 3 3‐sulphate in human plasma was completed using reversed phase high performace 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.