Analysis of the metabolites of genistin and enzyme kinetics of genistin in rat liver microsome
- Author:
Pei-Pei JIA
1
Author Information
- Publication Type:Journal Article
- Keywords: Enzymatic kinetics; Genistein; Genistin; HPLC-MS; Liver microsomes
- From: Chinese Pharmaceutical Journal 2015;50(9):797-801
- CountryChina
- Language:Chinese
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Abstract:
OBJECTIVE: To characterize the metabolism of genistin and study its enzymatic kinetics in rat liver microsome by HPLC-MS. METHODS: Genistin was incubated with rat liver microsomal incubation system. HPLC-MS method was used to characterize the metabolites. A metabolite generation method was established for quantitative analysis of genistein with sulfamethoxazole as internal standard. The enzyme kinetics parameters V
max and Km was calculated by the GraphPad Prism 5.0 software. RESULTS: The metabolites in vitro incubation system was identified as genistein. The optimal time in rat liver microsomes incubation time of 40 min, the optimal protein concentration of 1 mg · mL-1, a substrate concentration of 50 μmol · L-1. The enzyme kinetics parameters of genistein were as follows; Vmax =(0.1042 ± 0.0033) μmol · min-1 · mg(pro)-1, Km =(28.96 ± 2.80) μmol · L-1. CONCLUSION: The results indicate that genistin can be metabolited as the form of hydroxylation in rat liver microsome. Metabolite generation method is a reliable and simple method for determination of kinetic parameters of hepatic microsomal enzymes, and enzyme kinetic parameters obtained of genistin provide important parameters for further study.
