Establishment of Fingerprint and Spectrum-effect Relationship Study on Anti-oxidantion Activity of Anemarrhena asphodeloides and Its Different Processed Products / 中国药房

ABSTRACT

OBJECTIVE：To establish the fingerprint of ethanol extract and acetone extract from Anemarrhena asphodeloides and its different processed products ，and to investigate the spectrum-effect relationship between the fingerprint and the antioxidant activity. METHODS ：HPLC method and HPLC-ELSD method were adopted. The determination was performed on Thermo BDS Hypersil C 18 column with mobile phase consisted of acetonitrile- 0.2％ acetic acid at the flow rate of 1.0 mL/min. The column temperature was 30 ℃，and the detection wavelength was set at 258 nm. The sample size was 10 μL. The determination was performed on XDB-C 18 columnwith mobile phase consisted of acetonitrile-0.1％ acetic acid （gradient elution ）at the flow rate of 0.9 mL/min. The column temperature was 30 ℃ . The temperature of atomizer was 40 ℃ and the flow rare of N 2 was 1.6 mL/min. The sample size was 10 μL. Using mangiferin and timosaponin B Ⅱ as reference ，Fingerprint Similarity Eva- com luation System of TCM Chromatogram （2004A edition ）was adopted to draw the fingerprint of ethanol extract and acetoneextract from 20 batches of A. asphodeloides and its different processed products to confirm common peaks. Using scave nging rate of 1，1-diphenyl-2-trinitrophenylhydrazine（DPPH）radical as index，antioxidant activities of ethanol extract and acetone extract from 20 batches of A. asphodeloides and its processed products were investigated. Using scavenging rate of DPPH radical as dependent variable ，common peak area as independent variable ，PLSR was used to analyze the spectrum-effect relationship of ethanol extract and acetone extract from A. asphodeloides with antioxidantion activity. RESULTS ：Eight peaks （M1-M8）were identified in the fingerprints of ethanol extracts from 20 batches of processed A. asphodeloides . Mangiferin （chromatogram peak M 7）was identified with similarity of 0.389-1.000；seven comon peaks （S1-S7）and timosaponin B Ⅱ（peak S 5）were identified in the fingerprint of acetone extract ，and the similarity was 0.044-0.999. DPPH radical scavenging rate of ethanol extract from 20 batches of A. asphodeloides and its processed products was 21.23％- 81.39％，and A. asphodeloides was significantly lower than salt-processed A. asphodeloides with salt wine-processed A. asphodeloides （P＜0.001）；and that of acetone extract was 49.73％-83.78％，and A. asphodeloides was significantly higher than stir-baked A. asphodeloides with salt ，wine or fire （P＜0.001）. The standardized regression coefficients of peaks M 2-M7 in the spectrum of ethanol extract from A. asphodeloides were all greater than 0，which was positively correlated with antioxidant activity. Only the variable importance projection （VIP）value of peak M 7 was greater than 1，which had an important contribution. The standardized regression coefficients of peaks S 4-S7 in the acetone extract spectrum of A. asphodeloides were greater than 0，and were positively correlated with antioxidant activity. The order of VIP values was peak S 5＞S6＞S4，and the VIP values were all greater than 1. CONCLUSIONS：The fingerprint of the different processed products A. asphodeloides and its antioxidant activity spectral effect relationship were successfully established ；mangiferin（peak M 7）may be the main antioxidant substance of ethanol extract from A. asphodeloides . Timosaponin B Ⅱ（peak S 5），peak S 6 and peak S 4 may be the main antioxidant substance in acetone extract from A. asphodeloides .