HPLC determination of five polyphenols in rat plasma after intravenous administration of hawthorn leaves extract and its application to pharmacokinetic study.
A simple and specific HPLC-UV method was developed to simultaneously determine five active compounds including vitexin-4"-O-glucoside (VG), vitexin-2"-O-rhamnoside (VR), vitexin (VIT), rutin (RUT) and hyperoside (HP) in rat plasma after intravenous administrating the hawthorn leaves extract (HLE). With baicalin as internal standard (I.S.), sample pretreatment involved a one-step extraction with methanol of 0.2 ml plasma. The HPLC assay was carried out using a Phenomsil C18 analytical column with UV detection at 332 nm. The mobile phase consisted of methanol-acetonitrile-tetrahydrofuran-1% glacial acetic acid (6:1.5:18.5:74, v/v/v/v). The calibration curves were liner over the range of 2.030-500.5, 0.1513-75.64, 0.2507-12.54, 0.5128-25.64 and 0.4032-20.16 µg/ml for VG, VR, VIT, RUT and HP, respectively. The relative standard deviations (RSD) of the intra- and inter-day precisions for the analysis of the five analytes were between 1.0 and 8.9% with accuracies (relative error) below 8.2% for the analysis of the five analytes. The average extraction recoveries of five analytes were more than 82.67 ± 4.74%. The HPLC method herein described was fully validated and successfully applied to the pharmacokinetic studies after intravenous administration of HLE solution to rats over three doses.
Department of Pharmaceutical Analysis, School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian, China.
Si-Yuan
SY
Wang
Ji-Yan
JY
Chai
Wen-Jie
WJ
Zhang
Xun
X
Liu
Yang
Y
DU
Zhong-Zhe
ZZ
Cheng
Xi-Xiang
XX
Ying
Ting-Guo
TG
Kang
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D027825:Crataegus; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D005419:Flavonoids / Q000097:blood* / Q000737:chemistry; D007275:Injections, Intravenous; D008297:Male; D010636:Phenols / Q000097:blood* / Q000737:chemistry; D018515:Plant Leaves; D059808:Polyphenols; D051381:Rats; D017208:Rats, Wistar
D004365:Drugs, Chinese Herbal; D005419:Flavonoids; D010636:Phenols; D059808:Polyphenols