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期 刊 ：中国中药杂志 2008年 33卷 23期 页码：0.

关键词：葛根素；大豆苷元；HPLC- ESI-MS；肝微粒体；体外代谢；

Keywords:puerarin, daidzein, HPLC-MS, liver microsomes, metabolism in vitro,

摘 要 ：目的：建立Beagle犬肝微粒体中葛根素及其代谢物的液相色谱质谱测定方法，并对葛根素在Beagle犬肝微粒体中的代谢动力学进行研究。方法：超速离心法制备Beagle犬肝微粒体；Shimadzu-ODS色谱柱(2.0 mm×150 mm，5 μm)；流动相甲醇-水梯度洗脱，正离子模式，选择离子扫描m/z 417(M+H)+(葛根素)，m/z 531(2M+Na)+(大豆苷元)。结果：在体外代谢系统中，葛根素在Beagle犬肝微粒体中被代谢为大豆苷元，其酶促动力学参数Vmax为(0.047±0.006) mg·min-1·g-1，Km为(1.22±0.53) mg·L-1。结论：该HPLC-MS法能够准确灵敏测定犬肝微粒体中葛根素和大豆苷元；葛根素在Beagle犬肝微粒体中被P450酶代谢为大豆苷元，同时为白葛胶囊后续研究奠定基础。

Abstract:Objective: To develop a HPLC-ESI-MS method for the determination of puerarin and its metabolite and study the metabolic kinetics in beagle dog liver microsomes. Method: Beagle dog liver microsomes were prepared by using ultracentrifugation method. Chromatography was performed on a Shimadzu C18 column (2. 0 mm×150 mm，5 μm). Amethanol-water gradient system was used. ESI interface was applied in the positive, and SIM m/z 417 was puerarin and m/z 531 was daidzein. Result: The puerarin was metabolized by NADPH regenerating system in beagle dog microsomes. The Michaelis-Menten parameters Km and Vmax in beagle dog microsomes were initially estimated by analyzing Lineweave-Brurk plot. The Vmax, Km of puerarin were (0.047±0.006) mg·min-1·g-1, (1.22±0.53) mg·L-1. Conclusion: The puerarin and daidzein can be rapidly determined by HPLC-MS in beagle dog microsomes and the puerarin was metabolized to daidzein by CY P450. The study can give help for Baige capsule.

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