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Simultaneous Determination of Baicalein, Baicalin, Wogonin, and Wogonoside in Rat Plasma by LC-MS/MS for Studying the Pharmacokinetics of the Standardized Extract of Scutellariae Radix

  • Chung, Hye-Jin (Neuro-Medicine Center, Korea Institute of Science and Technology) ;
  • Lim, Sun-Young (College of Pharmacy, Hanyang University) ;
  • Kim, In-Sook (College of Pharmacy, Hanyang University) ;
  • Bu, Young-Min (Department of Herbal Pharmacology, College of Oriental Medicine, Kyung Hee University) ;
  • Kim, Ho-Cheol (Department of Herbal Pharmacology, College of Oriental Medicine, Kyung Hee University) ;
  • Kim, Dong-Hyun (Department of Pharmacology and PharmacoGenomics Research Center, College of Medicine, Inje University) ;
  • Yoo, Hye-Hyun (College of Pharmacy, Hanyang University)
  • Received : 2011.10.21
  • Accepted : 2011.11.17
  • Published : 2012.01.20

Abstract

A new composition of standardized Scutellariae Radix extract (HPO12) was developed for treatment of Alzheimer's disease. For the preclinical pharmacokinetic study of HPO12, a rapid, sensitive, and selective LCMS/MS method was developed and validated for the simultaneous determination of 4 bioactive compounds, baicalein, baicalin, wogonin, and wogonoside. After extraction with ethylacetate, chromatographic analysis was performed on a Thermo $C_{18}$ column ($150mm{\times}2.1mm$, $3{\mu}m$) with a mobile phase consisting of 0.1% formic acid (A) and 0.1% formic acid in 95% acetonitrile (B) by using gradient elution at a flow rate of $250{\mu}L/min$. Analytes introduced to a mass spectrometer were monitored by multiple reaction monitoring (MRM) in positive ion mode. Using $25{\mu}L$ of plasma sample, the method was validated over the following concentration ranges: 25-5000 ng/mL for baicalein, 20-40000 ng/mL for baicalin, 1-1000 ng/mL for wogonin, and 5-10000 ng/mL for wogonoside. The intra- and inter-day precision and accuracy of the quality control samples at the 4 concentrations showed $\leq$ 13.7% relative standard deviation (RSD) and 86.6-105.5% accuracy. The method was successfully applied to determine the concentrations of baicalein, baicalin, wogonin, and wogonoside in rat plasma after intraperitoneal and oral administrations of HPO12.

Keywords

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