Research Article

Simultaneous determination of tanshinone IIA and its three hydroxylated metabolites by liquid chromatography/tandem mass spectrometry

  1. Peng Li1,
  2. Guang-Ji Wang1,*,
  3. Jing Li1,2,
  4. Hai-Ping Hao1,
  5. Chao-Nan Zheng1

Article first published online: 7 FEB 2006

DOI: 10.1002/rcm.2367

Issue

Rapid Communications in Mass Spectrometry

Rapid Communications in Mass Spectrometry

Volume 20, Issue 5, pages 815–822, 15 March 2006

Additional Information

How to Cite

Li, P., Wang, G.-J., Li, J., Hao, H.-P. and Zheng, C.-N. (2006), Simultaneous determination of tanshinone IIA and its three hydroxylated metabolites by liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom., 20: 815–822. doi: 10.1002/rcm.2367

Author Information

  1. 1

    Key Lab of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210038, China

  2. 2

    Department of Pharmacology, School of Medicine, Jiangsu University, Zhenjiang 212001, China

*Key Lab of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210038, China.

Publication History

  1. Issue published online: 7 FEB 2006
  2. Article first published online: 7 FEB 2006
  3. Manuscript Revised: 31 DEC 2005
  4. Manuscript Accepted: 31 DEC 2005
  5. Manuscript Received: 25 NOV 2005

Funded by

  • National ‘863’ Project. Grant Number: 2003AA2Z347A
  • Program in International Cooperation. Grant Number: BZ2004042
  • Key Lab of Drug Metabolism and Pharmacokinetics of Jiangsu Province. Grant Number: BM2001201

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Abstract

A rapid and sensitive method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the simultaneous determination of tanshinone IIA and its three hydroxylated metabolites, tanshinone IIB, hydroxytanshinone IIA and przewaquinone A, in a rat liver microsome was developed and fully validated. A single step of liquid-liquid extraction with ethyl acetate was utilized in this method. Chromatographic separation of the sample matrix from the analytes and the internal standard diazepam was performed using a Shim-pack VP-ODS analytical column. Detection was performed on a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization source and operated in selected reaction monitoring (SRM) mode. The method was linear in the concentration range of 1–500 ng/mL for all analytes. The intra- and inter-day precisions (RSD %) were within 15% and deviations of the assay accuracies were within 15.0% for all analytes. The analytes proved to be stable during sample storage, preparation and analyses. This validated method was successfully applied to the enzyme kinetic study of tanshinone IIA in liver microsome. The elimination of tanshinone IIA and formation of tanshinone IIB and hydroxytanshinone IIA in the liver microsome all exhibited a sigmoidal kinetics profile. The formation of przewaquinone A shows a typical hyperbolic profile. In addition, this method has now been applied in the analysis of other bio-samples including plasma, urine, bile and feces. Copyright © 2006 John Wiley & Sons, Ltd.

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