Abstract: Aconite tuber is one of the commonly used Chinese traditional medicine. Lipo-alkaloids is the most important active ingredients in Aconite tuber. A stable, efficient and reliable method was established to quickly analysed the lipo-alkaloids which are rather complex in aconite tuber. The lipo-alkaloids in aconite tuber is analysed by using multiple scan function of electrospray ionization coupled with triple quadrupole mass spectrometry including full scan, daughter ion scan, parent ion scan and neutral loss scan. There are two kinds of differences in the lipo-alkaloidal structures, one in which base on the different alkaloid skeleton, the other difference is due to the connected with C8 substituted long chain fatty acids. The lipo-alkaloids with the high content in aconite tuber are due to 4 kinds of alkaloids skeleton and 3 kinds of C8 substituted fatty acids, e.g. a total of 12 kinds of lipo-alkaloids which are analysed. People often only focus on the much reaction monitoring(MRM) function of the triple quardrupole mass spectrometry, ignore the other functions. The function of parent ion scan and neutral loss scan for analysis of the lipo-alkaloids from aconite root has scanning speed, high efficiency. The results are stable and reliable.