Separation and characterization of monoglucuronides of vitamin D(3) and25-hydroxyvitamin D(3) in rat bile by high-performance liquid chromatography.
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 beta-glucuronidase.
Shimada
K
K
Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Nakatani
I
I
Saito
K
K
Mitamura
K
K
D000818:Animals; D001646:Bile / Q000737:chemistry*; D002112:Calcifediol / Q000031:analogs & derivatives*; D002762:Cholecalciferol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005456:Fluorescent Dyes / Q000378:metabolism; D005965:Glucuronates / Q000737:chemistry* / Q000302:isolation & purification; D005966:Glucuronidase / Q000378:metabolism; D006868:Hydrolysis; D008297:Male; D015394:Molecular Structure; D051381:Rats; D017208:Rats, Wistar
D005456:Fluorescent Dyes; D005965:Glucuronates; C027963:vitamin D3 glucosiduronate; D002762:Cholecalciferol; D005966:Glucuronidase; D002112:Calcifediol