[Metabolic kinetic of puerarin in beagle liver microsomal by HPLC-ESI-MS].
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.


METHODS
Beagle dog liver microsomes were prepared by using ultracentrifugation method. Chromatography was performed on a Shimadzu C18 column (2.0 mm x 150 mm, 5 microm). 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.


RESULTS
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 x min(-1) x g(-1), (1.22 +/- 0.53) mg x L(-1).


CONCLUSIONS
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.
Wen
Bin-yu
BY
Xi'an Jiaotong University School of Medicine, Xi'an 710061, China. wenbinyu@stu.xjtu.edu.cn
Li
Hao
H
Wang
Lan
L
Wang
Si-cen
SC
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D007529:Isoflavones / Q000493:pharmacokinetics*; D008099:Liver / Q000737:chemistry* / Q000187:drug effects; D008862:Microsomes, Liver / Q000737:chemistry* / Q000187:drug effects; D010599:Pharmacokinetics; D021241:Spectrometry, Mass, Electrospray Ionization
D007529:Isoflavones; C033607:puerarin