Study on Specific Chromatogram and Multi-components Determination of Alkaloids in Hulisan Tablets by HPLC

OBJECTIVE To establish the HPLC specific chromatogram of aconitum alkaloids in Hulisan tablets and simultaneously determine the contents of 6 alkaloids. METHODS Kromasil 100-5-C₁₈ column(4.6 mm×250 mm, 5 μm) was used with gradient elution of acetonitrile(A)-0.2% acetic acid(triethylamine adjusted pH to 6.20)(B) at the flow rate of 1.0 mL·min^-1. The column temperature was 35℃ and the detection wavelength was 235 nm. The similarity evaluation system for chromatographic fingerprint of TCM was used to calculate the similar degree. Then, the further quality assessment of the drug was carried out by similarity evaluation combined with CA and PCA. And the contents of the active ingredients of sample were determined. RESULTS Ten common peaks were identified in the aconitum alkaloids characteristic chromatogram of 11 batches of Hulisan tablets. And 6 alkaloids were identified as benzoylmesaconitine, benzoylhypaconitine, benzoylaconitine, mesaconitine, hypaconitine and aconitine. The contents of benzoylmesaconitine, benzoylhypaconitine, benzoylaconitine, mesaconitine, hypaconitine and aconitine in 11 batches of Hulisan tablets from three manufacturers were 231.190-1 002.977, 17.022-105.744, 21.962-48.406, 5.436-39.678, 2.692-97.854 and 1.707-43.342 μg·g^-1. CONCLUSION The established HPLC specific chromatogram method and multi-component content determination method is simple, reliable, accurate and can effectively evaluate the quality of Hulisan tablets.