Abstract

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 µm 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.

Key words: Aconitum alkaloids, rat body fluids, quantitative analysis, high-performance liquid chromatography.

Abbreviation

HPLC, High-performance liquid chromato- graphy; DAD, photodiode array detector; RSD, relative standard deviations; SPE, solid phase extraction; LC-MS, liquid chromato- graphy-mass spectrometry; GC-MS, gas chromatography-mass spectrometry; CE, capillary electrophoresis; CE-MS,capillary electrophoresis-mass spectrometry.