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

• Published in: Biological & pharmaceutical bulletin Vol. 19; no. 4; pp. 491 - 494
• Main Authors: SHIMADA, Kazutake, NAKATANI, Ito, SAITO, Kyoko, MITAMURA, Kuniko
• Format: Journal Article
• Language: English
• Published: Tokyo The Pharmaceutical Society of Japan 1996; Maruzen; Japan Science and Technology Agency
• Subjects: 25-hydroxyvitamin D3; biliary metabolite; Biological and medical sciences; Enzymes. Coenzymes. Vitamins. Pigments; Fundamental and applied biological sciences. Psychology; identification; Metabolisms and neurohumoral controls; monoglucuronide; rat; Vertebrates: anatomy and physiology, studies on body, several organs or systems; vitamin D3; Separation; Rat; Metabolite; Rodentia; HPLC chromatography; Identification; Glucuroconjugate; Characterization; Vertebrata; Mammalia; Vitamin D; Animal; Cholecalciferol(25-hydroxy); Bile; Colecalciferol
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.19.491

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.