Simultaneous determination of nine major constituents in Agrimonia pilosa Ledeb. by HPLC-DAD-ESI-MS/MS

Herba Agrimoniae is a traditional Chinese medicine. To date, there is no quantitative control method for Agrimonia pilosa (A. pilosa) with main active constituent contents listed in the Chinese Pharmacopeia. A high performance liquid chromatography coupled with diode-array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS/MS) method is established for the characterization and simultaneous quantification of nine major constituents in A. pilosa. Furthermore, the contents of these constituents of A. pilosa from 21 different regions of China were analyzed and compared. All calibration curves show good linearity (r ≥ 0.999). The HPLC-DAD method shows good precision, repeatability and stability for the quantification of the nine constituents in A. pilosa with the standard deviations below 3.29%. The recoveries for the quantified constituents are within the range of 95.44–102.55%. The qualitative and quantitative method was proved to be sensitive, accurate and reliable to determine the constituents in A. pilosa. The method is successfully applied to differentiate the A. pilosa of different regions.