Se Pu. 2002 May; 20(3): 253-5.

[Determination of aconitine alkaloids in a pesticide--0.25% aconitine alkaloids mixed with emulsion by high performance liquid chromatography]

Chen YF, He W, Zhu FC.

Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.

The raw root of Aconitum, an important Chinese traditional medicine, contains some very toxic alkaloids such as aconitine, mesaconitine and hypaconitine etc. They are usually processed to lower the alkaloid content before used as a drug. The extract of crude drug (aconite) can be made into a plant pesticide by means of mixing with some emulsion. In order to evaluate the quality of the pesticide, we developed a rapid, specific and precise method using high performance liquid chromatography (HPLC) for the separation and quantitation of the alkaloids in the aconite extract and the mixed products (the aconite extract is mixed with emulsion). Before the determination by HPLC, the sample must be acidified with 2% (mass percentage) HCl at first. Extract the acid liquid with CHCl3. Alkalize the extract with ammonia, and add a little of 0.1 mol/L NaHCO3 and 0.1 mol/L Na2CO3 till pH 9. Extract it with CHCl3. Evaporate the extract and add a certain amount of methanol. Add the internal standard into the sample. Inject the combined sample solution onto a column of chemically bonded octadecylsilane phase and develop the chromatogram with MeOH-H2O-CHCl3-triethylamine (68:32:2:0.1, volume ratio). Aconitine, mesaconitine, hypaconitine and medroxyprogesteroni acetas (internal standard) were on base line separated. Quantify the alkaloids by peak area ratio (aconitine alkaloids vs internal standard). This method has high recovery (> 92%) and good reproducibility (RSD < 3.2%).