Objective: To establish a liquid chromatography-mass spectrometry-mass spectrometry (LC-MS-MS) method to study the effects of Phellodendri Cortex (CP) on the contents of four ingredients (mangiferin

neomangiferin

timosaponin AⅢ and timosaponin BⅡ) from Anemarrhenae Rhizoma (RA). Method: Chromatographic separation was carried out on ZORBAX Eclipse XDB-C18 column (2.1 mm×150 mm

3.5 μm)

with 0.1% formic acid-acetonitrile as the mobile phase for gradient elution. The column temperature was 30 ℃ and the flow rate was 0.25 mL ·min-1. The quantification of the analytes was performed by mass spectrometry with electrospray ionization (ESI) under negative ion multiple reaction monitoring (MRM) mode. Result: Mangiferin

neomangiferin

timosaponin AⅢ and timosaponin BⅡhad good linear relationship between 5.05-404

5.3-424

5.05-404

20.8-1 664 μg ·L-1 respectively (r ≥0.999). The average recovery rate was 100.6%

99.2%

99.1% and 98.5%

respectively. The contents of mangiferin and timosaponin AⅢ were highest in group RA-CP (1: 0). Neomangiferin content in group RA-CP (1: 1) was higher than that in other groups. Timosaponin BⅡcontent was similar in each group. Conclusion: The method is rapid

accurate and reliable

and has been successfully used to the quantification of four compounds above mentioned in RA-CP. The contents mangiferin

neomangiferin and timosaponin AⅢ can be affected by the different RA-CP compatibility.