Simultaneous determination of tanshinone IIA and its three hydroxylated metabolites by liquid chromatography/tandem mass spectrometry
Article first published online: 7 FEB 2006
Copyright © 2006 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 20, Issue 5, pages 815–822, 15 March 2006
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
- Issue published online: 7 FEB 2006
- Article first published online: 7 FEB 2006
- Manuscript Revised: 31 DEC 2005
- Manuscript Accepted: 31 DEC 2005
- Manuscript Received: 25 NOV 2005
- 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
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.