Separation and Characterization of Monoglucuronides of Vitamin D_3 and 25-Hydroxyvitamin D_3 in Rat Bile by High-Performance Liquid Chromatography

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The separation and characterization of vitamin D_3- and 25-hydroxyvitamin D_3-monoglucuronides, biliary metabolites obtained from rats dosed with D_3 and 25-hydroxyvitamin D_3 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.
公益社団法人日本薬学会の論文
1996-04-15