经典名方身痛逐瘀汤物质基准UPLC指纹图谱建立及其5种成分含量测定

UPLC fingerprint and determination of five components of substance benchmark of classical prescription Shentong Zhuyu Decoction

采用Waters Sun Fire C_(18)色谱柱(3.0 mm×150 mm,3.5μm),以乙腈-0.1%磷酸水溶液为流动相,梯度洗脱,体积流量0.4 mL·min^(-1),采用多波长切换法,柱温35℃,对18批身痛逐瘀汤物质基准样品进样分析。建立了经典名方身痛逐瘀汤物质基准UPLC指纹图谱,共标定了16个共有峰,使用《中药色谱指纹图谱相似度评价系统》(2012版)计算相似度,其相似度在0.911~0.988,并结合聚类分析(cluster analysis,CA)、主成分分析(principal component analysis,PCA)以及偏最小二乘法判别分析(partial least squares discrimination analysis,PLS-DA)、化学计量法,以16个共有峰进行评价,18批身痛逐瘀汤物质基准被分为2类,样品S1、S2、S5~S8、S14、S17分为一类,样品S3、S4、S9~S13、S15、S16、S18分为一类。筛选出差异性较大的11个成分,对差异较大且已知的羟基红花黄色素A、阿魏酸、苯甲酸、蜕皮激素及甘草酸铵5种有效成分进行含量测定,结合指纹图谱可以反映身痛逐瘀汤物质基准的整体概貌。该研究建立的经典名方身痛逐瘀汤物质基准UPLC指纹图谱及多成分含量测定方法,简单可行,重复性、稳定性良好,可为身痛逐瘀汤物质基准质量控制的研究提供依据。

Fingerprints of 18 batches of substance benchmark of Shentong Zhuyu Decoction(SZD)were established by UPLC under the following conditions:Waters Sun Fire C_(18) column(3.0 mm×150 mm,3.5μm),column temperature of 35℃,gradient elution with mobile phase of acetonitrile(A)-0.1%phosphoric acid aqueous solution(B)at the flow rate of 0.4 mL·min^(-1),and detection by wavelength switching.A total of 16 common peaks were identified.The similarities among the fingerprints were calculated by Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Edition)and the result showed they were in the range of 0.911-0.988.Based on the 16 common peaks,cluster analysis(CA),principal component analysis(PCA),and partial least square discriminant analysis(PLS-DA)all categorized the 18 batches of samples into two groups(S1,S2,S5-S8,S14,and S17 in one group,and S1,S2,S5-S8,S14,and S17 in another),and 11 most influential components were screened.Five known components with great difference among samples(hydroxysafflor yellow A,ferulic acid,benzoic acid,ecdysone,and ammonium glycyrrhizinate)were determined.The combination of multi-component content determination and fingerprints can reflect the overall cha-racteristics of the primary standards of SZD,which is simple,feasible,reproducible,and stable.This study can serve as a reference for the quality control of the primary standards of SZD.