On-Line Drug Metabolism in Capillary Electrophoresis. 1. Glucuronidation Using Rat Liver Microsomes
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- 作者：Hee Seung Kim ; Irving W. Wainer
- 刊名：Analytical Chemistry
- 出版年：2006
- 出版时间：October 15, 2006
- 年：2006
- 卷：78
- 期：20
- 页码：7071 - 7077
- 全文大小：123K
- 年卷期：v.78,no.20(October 15, 2006)
- ISSN：1520-6882
文摘
A rat liver microsome pseudostationary phase has beenused for the on-line capillary electrophoresis monitoringof glucuronidation. Uridine diphosphate glucuronosyltransferase (EC 2.4.1.17) containing microsomes wasisolated from rat liver and directly injected onto neutrallycoated capillary containing polymeric replaceable gelsfollowed by injection of the substrate mixture. On-lineglucuronidation was observed within 15 min without anysample preparation. The factors affecting the separationof glucuronides and parent compounds were investigatedby varying the applied electric fields and the size (lengthand internal diameter) of capillary. The Michaelis-Mentenparameters (Km and Vmax) for the glucuronidation of4-methyl-7-hydroxy coumarin and 4-nitrophenol weredetermined using the CE method and by off-line microsomal incubation. No significant differences were observedfor Km and Vmax values for 4-methyl-7-hydroxycoumarinand 4-nitrophenol between on-line and off-line glucuronidation of these two compounds. This method was alsoused to determine the inhibition constant (IC50 value) forthe competitive inhibition of morphine glucuronidation bycodeine, IC50 (on-line) = 170 vs 580 
M (off-line). Theresults demonstrate that this method can be used toscreen for the glucuronidation of test compounds andshould reduce the time required for this screening process.
M (off-line). Theresults demonstrate that this method can be used toscreen for the glucuronidation of test compounds andshould reduce the time required for this screening process.