Comparison of Component Differences in Chebulae Fructus and Chebulae Fructus Immaturus Based on Chemical Pattern Recognition and Multi-index Quantitative Analysis / 中国药房

ABSTRACT

OBJECTIVE：To provide reference for the identification of Chebulae Fructus and Chebulae Fructus Immaturus . METHODS：UPLC method was adopted. The determination was performed on Waters Cortecs T 3 C18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid solution （gradient elution ）at the flow rate of 0.35 mL/min. The column temperature was 30 ℃，and the detection wavelength was set at 270 nm. The sample size was 1 μL. Using gallic acid as reference，UPLC fingerprints of 17 batches of Chebulae Fructus and 14 batches of Chebulae Fructus Immaturus were established and their similarity was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition）. By comparing substance control ， UV absorption spectrum and related literaturs ，common peaks were identified. PCA and PLS-DA were performed by using SPSS 20.0 and SIMCA 14.1 software. The contents of main difference components in Chebulae Fructus and Chebulae Fructus Immaturus were determined by above UPLC method and compared. RESULTS ：There were 8 common peaks in UPLC fingerprint of Chebulae Fructus and Chebulae Fructus Immaturus ，i.e. chebulic acid （peak 1），gallic acid （peak 2），punicalagin A （peak 3），punicalagin B （peak 4），corilagin（peak 6），chebulagic acid （peak 7）and chebulinic acid （peak 8）. The similarities of 17 batches of Chebulae Fructus were from 0.92 to 0.99，while 14 batches of Chebulae Fructus Immaturus were all above 0.99. The similarity of control fingerprint between Chebulae Fructus and Chebulae Fructus Immaturus was 0.909. PCA demonstrated the differences between Chebulae Fructus and Chebulae Fructus Immaturus . The results of PLS-DA were consistent with those of PCA ，and the variable importance in projection （VIP）values of peak 5，4，7，3 and 2 were above 1 in the PLS-DA model. In 31 batches of samples ，the contents of gallic acid （peak 2），punicalagin A（peak 3），punicalagin B （peak 4）and chebulagic acid （peak 7）were 2.63-10.31， 5.37-44.63，8.02-60.77，44.07-162.98 mg/g；RSDs were 40.14％， 47.91％ ，53.97％ ，36.22％（n＝31）. There was statistical significance in the differences of the mentioned 4 components between Chebulae Fructus and Chebulae Fructus Immaturus 719412818@qq.com （P＜0.05）. CONCLUSIONS ：There are significant differences between Chebulae Fructus and Chebulae Fructus Immaturus gallic acid ，punicalagin A ，punicalagin B and chebulagic acid are the main difference components for identification.