Separation and Characterization of Monoglucuronides of Vitamin D₃ and 25-Hydroxyvitamin D₃ in Rat Bile by High-Performance Liquid Chromatography

抄録

The separation and characterization of vitamin D₃- and 25-hydroxyvitamin D₃-monoglucuronides, biliary metabolites obtained from rats dosed with D₃ and 25-hydroxyvitamin D₃ per os, respectively, were carried out by HPLC. The glucuronide fractions were obtained from bile specimens by the combined use of a Bond Elut C18 cartridge, for solid phase extraction, and a lipophilic gel (piperidinohydroxypropyl Sephadex LH-20), for ion-exchange chromatography. Each glucuronide was identified by comparison with an authentic sample in three ways : its chromatographic behavior, that of its fluorescent labeled derivative using 4-[4-(6-methoxy-2-benzoxazolyl)phenyl]-1, 2, 4-triazoline-3, 5-dione and data obtained following enzymatic hydrolysis using β-glucuronidase.