QUANTITATIVE-DETERMINATION OF 25-HYDROXYVITAMIN D-3 3-SULFATE IN HUMAN PLASMA USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

The quantitative determination of 25-hydroxyvitamin D-3 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 ngimL. The proposed method is satisfactory in its accuracy and precision.