Shimada K, Nakatani I, Saito K, Mitamura K
Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Biol Pharm Bull. 1996 Apr;19(4):491-4. doi: 10.1248/bpb.19.491.
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.
分别通过口服给予维生素D₃和25-羟基维生素D₃的大鼠所获得的胆汁代谢产物——维生素D₃和25-羟基维生素D₃单葡萄糖醛酸苷的分离与表征，采用高效液相色谱法（HPLC）进行。通过联合使用用于固相萃取的Bond Elut C18柱和用于离子交换色谱的亲脂性凝胶（哌啶羟基丙基葡聚糖凝胶LH-20），从胆汁标本中获得葡萄糖醛酸苷级分。通过三种方式与标准样品比较来鉴定每种葡萄糖醛酸苷：其色谱行为、使用4-[4-(6-甲氧基-2-苯并恶唑基)phenyl]-1,2,4-三唑啉-3,5-二酮的荧光标记衍生物的色谱行为，以及使用β-葡萄糖醛酸苷酶进行酶促水解后获得的数据。
