Abstract: UFLC-Qtrap-MS/MS method was established to simultaneously determine 11 kinds of free amino acids and 11 kinds of nucleosides in Changium smyrnioides Wollf and its in vitro cultures. Waters XBridge Amide column(2.1 mm×100 mm,3.5 μm)was used for chromatographic separation at 30 ℃. The flow rate was 0.6 mL·min^-1 with a mobile phase of 0.2% formic acid-0.2% formic acid in acetonitrile in gradient elution mode. The compounds were detected by positive ion multiple reaction monitoring mode. The results were analyzed by principal component analysis(PCA)and technique for order preference by similarity to an ideal solution(TOPSIS). The results showed that,relative standard deviations of precision,repeatability,stability and recovery tests were all less than 5%. The average recovery values were within 98.62%~100.64%. The linear relationship of measured components was good,which showed that the method was accurate and feasible. The contents of free amino acids and nucleosides in different samples varied greatly. The suspension cells from petiole contained the highest content of total free amino acids(3682.01 μg·g^-1)and total nucleosides(6241.90 μg·g^-1). The highest evaluation index of amino acids was in cultivated C. smyrnioides Wollf root while that of nucleosides was in suspension cells cultured from petiole. TOPSIS evaluation showed that the quality of petiole suspension cell was the best. The method established was suitable for simultaneous and rapid determination of 22 kinds of free amino acids and nucleosides in C. smyrnioides Wollf. The quality evaluation pointed out the utility value of the root,leaf,petiole suspension cell and regenerated leaf of C. smyrnioides Wollf. The study provided some options for rational utilization of C. smyrnioides Wollf resource.