Simultaneous determination of aconitum alkaloids in rat body fluids by high-performance liquid chromatography

A sensitive, reliable and accurate high-performance liquid chromatography (HPLC) method coupled with photodiode array detector (DAD) were developed for the simultaneous quantitative determination of aconitine, mesaconitine and hypaconitine in rat plasma and urine by optimizing the extraction, separation and analytical conditions. The analyses were chromatographed on a ZORBAX Eclipse XDB-C18 column (150 mm ~ 4.6 mm i.d.; 5 ƒEm particle size) with gradient elution using solvents of acetonitrile and ammonium acetate buffer (pH 10.0). The detection wavelength was 240 nm. Intra-assay and inter-assay precision of the analyses were less than 10% and the average recovery rates obtained were in the range of 85.63 - 90.94% for all analysis of the three aconitum alkaloids with relative standard deviations (RSD) below 14%. Positive linear relationships were observed in correlation coefficients that exceeded 0.95. The limits of detection (signal-to-noise ratio of 3) were 2.64, 1.58 and 2.75 ng for aconitine, mesaconitine and hypaconitine, respectively. The method can provide a scientific and technical platform to determine the concentration of aconitum alkaloids in plasma during a pilot pharmacokinetic study in rats.