LC MS/MS determination and pharmacokinetic study of columbianadin in rat plasma after intravenous administration of pure columbianadin

Background: Columbianadin, one of the active coumarins, is isolated from Radix Angelicae pubescentis which has been used as a traditional Chinese medicine for the treatment of rheumatic diseases for thousands of years. A fast and sensitive method is required for the determination of columbianadin for pharmacokinetic studies. Liquid chromatography tandem mass spectrometry (LC-MS/MS) method is a preeminent analytical tool for rapid biomedical analysis. Results: A sensitive LC-MS/MS method has been validated to determine the concentration of columbianadin in rat plasma after intravenous administration of columbianadin (1, 2.5 and 5 mg kg(-1)). Liquid-liquid extraction was used to extract columbianadin from the rat plasma. Bergapten was selected as an internal standard (IS). The separations were performed on an Eclipse plus C18 column (4.6 100 mm, 1.8 mu m) with ammonium acetate aqueous solution (1 mmol L-1) and acetonitrile as the mobile phase. The flow rate was set at 0.300 mL min(-1). Quantification was performed using multiple reaction monitoring (MRM) mode to monitor transitions of m/z 329.3 -> 229.3 for columbianadin and m/z 217.2 -> 202.2 for IS at positive ionization mode. The calibration curve was linear over the concentration range of 4 20000 ng mL(-1) with a correlation coefficient (r) of 0.996 or better. The precision of intra-and inter-batch assays ranged from 4.02 to 7.33% and accuracies determined at three concentrations ranged between 91.9% and 106%. The lower limit of quantification was about 4 ng mL-1. Conclusion: The proposed LC-MS/MS method is simple, rapid and highly sensitive so that it could be used to evaluate pharmacokinetic properties of columbianadin in rat plasma after intravenous administration.