Abstract: A multiple indexes quantitative analysis method using ultra performance liquid chromatography united triple quadrupole mass spectrometry in multiple reaction monitoring (MRM) for determination of alkaloid content in Radix Aconitum Vilmoriniani was established. The research provide a reference for the quality control of Radix Aconitum Vilmoriniani. Chromatography columns: ACQUITY YPLC®BEH C₁₈ (ϕ2.1 mm×50 mm, 1.7 µm). Mobile Phases: acetonitrile (A)-0.1% formic acid aqueous solution (B), gradient elution. The contents of talatisamine, yunaconitine, vilmorrianine A and other six deterpenoid alkaloids in 18 batches of cultivated Radix Aconitum Vilmoriniani were determined by UPLC-MS/MS and evaluated combining with chemometric methods. Good linearity (R²>0.9980) was observed for the contents of nine ingredients with respect to their peak areas measured in the corresponding concentration ranges. The average spiked recovery rate were 96.74%−101.2% with the RSD of 0.65%−1.9%. The determination results showed that the contents of nine components in Radix Aconitum Vilmoriniani varied largely, but there were no significant geographical differences. According to the results of HCA, PCA and OPLS-DA, 18 batches of cultivated Radix Aconitum Vilmoriniani were found to be basically divided into two categories, in which talatismine,vilmorrianine A, yunaconitine, geniculatine C and austroconitine B were the main differential components. The method was developed for the simultaneous determination of the contents of nine alkaloid components in cultivated Radix Aconitum Vilmoriniani by UPLC-MS/MS. The method have advantage of high specificity and sensitivity with short analysis time, and can provide a reference for the quality evaluation of Radix Aconitum Vilmoriniani from different origins.