By optimizing the extraction and analytical conditions, a reliable and precise HPLC method coupled with photodiode array detection (HPLC-DAD) has been developed for the identification and quantification of three major aconitine-type alkaloids (aconitine, mesaconitine, hypaconitine) in the roots of Aconitum carmichaelii Debeaux. The qualitative analysis of the plant material was carried out by LC-APCI-MS(n). By means of this method, 26 lipo-alkaloids were also identified from the roots of A. carmichaelii. The effect of processing on aconitine-type alkaloids, lipo-alkaloids and pure aconitine was studied. As part of our investigation, two lipo-alkaloids, 14-benzoylaconine-8-palmitate and 14-benzoylaconine-8-linoleate were produced semisynthetically. The COX-1, COX-2 and LTB(4) formation inhibitory activity of aconite root extracts and different types of diterpene alkaloids and the toxicity of lipo-alkaloids were also investigated.