Determination of aconitum alkaloids in biological samples by liquid chromatography-tandem mass spectrometry

Objective:To establish a method for determination of aconitum alkaloids in biological samples by LC-MS/MS.Methods:After 0.5 mL(g) of biological sample was extracted using a liquid-liquid extraction with diethyl ether at pH 9.2 and buprenorphine as an internal standard,aconitum alkaloids were separated on a Capcell Pak MG-Ⅱ C18 column (250 mm×2.0 mm,5 μm) using acetonitrile-buffer solution(20 mmol·L-1 ammonium acetate and 0.1% formic acid,pH 4) (70∶30) as the mobile phase at a flow-rate of 0.2 mL·min-1 with LC-MS/MS in the positive ionization mode.Quantitative results were obtained by the internal calibration procedure.Results:Aconitine,hypaconitine,mesaconitine and buprenorphine in blood,urine and liver were separated well.Calibration curve was linear within the range of 0.1-50 ng·mL-1(ng·g-1) with the LOD range from 0.01 to 0.1 ng·mL-1(ng·g-1).The inter-day and intra-day RSDs were less than 13.3%.The extraction recoveries of aconitum alkaloids in blood were more than 64.8%.Conclusion:This method is sensitive,fast and accurate for determination of trace aconitum alkaloids in biological samples.