Objective To develop a HPLC-MS/MS method for simultaneous quantification of six alkaloids, namely aconitine, hypaconitine, mesaconitine, yunaconitine, bullatine A and bullatine B, in aconitum brachypodum in blood.
Methods The gradients were eluted with methanol-water with 0.1% formic acid as mobile phase and separated on Waters C18 column. The flow rate was 0.2 mL/min. Protein precipitation with acetonitrile was used in sample preparation. Electrospray ionization, multiple reaction monitoring scan mode were used for mass spectrometry detection.
Results The linear ranges were 0.5-100 ng/mL for six alkaloids. The limits of aconitine, hypaconitine, mesaconitine, yunaconitine, bullatine A and bullatine B were found to be 0.50, 0.26, 0.56, 0.03, 0.30 and 0.20 ng/mL, respectively. At three spiked concentrations, the average extraction recoveries ranged from 82.1%-90.3%, accuracy ranged from 98.9%-105.8%. The intra-day and inter-day RSDs were less than 7%.
Conclusions The method provides a sensitive, accurate, precise and reliable analytical procedure for clinical and toxicological monitoring of six alkaloids in aconitum brachypodum in whole blood.
DownLoad: