目的:建立栝楼桂枝汤(Gualou Guizhi decoction)物质基准指纹图谱和多成分含量测定方法,探究饮片-物质基准关键质量属性的量值传递规律.方法:制备15批栝楼桂枝汤物质基准,采用ChromCore 120 C18(250 mm × 4.6 mm,5 μm)色谱柱,以乙腈(A)-0.1％甲酸水(B)为流动相,进行梯度洗脱,流速0.8 mL·min-1,柱温30 ℃,检测波长254 nm,采用中药色谱指纹图谱相似度评价软件(2012年版)建立指纹图谱,结合层次聚类分析(hierarchical cluster analysis,HCA)、主成分分析(principal component an alysis,PCA)与正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA),筛选不同批次栝楼桂枝汤物质基准质量差异的主要化学成分.建立HPLC法测定主要差异性成分含量,以含量、出膏率和转移率分析栝楼桂枝汤的量值传递规律.结果:建立了栝楼桂枝汤物质基准HPLC指纹图谱,确定30个共有峰,与对照品比对共指认出其中9个成分,分别为没食子酸、芍药内酯苷、芍药苷、甘草苷、芒柄花苷、甘草素、肉桂酸、异甘草素、甘草酸.15批栝楼桂枝汤物质基准指纹图谱相似度均＞0.920.3种化学识别模式均将样品分为2类,造成主要差异化学成分共5个,分别为甘草酸、甘草素、芍药苷、甘草苷、芒柄花苷.建立了多成分HPLC含量测定方法,符合方法学要求.饮片-物质基准之间5个差异成分的平均转移率分别为51.65％、40.46％、74.74％、60.06％、34.54％,平均出膏率为14.20％.结论:栝楼桂枝汤物质基准的关键质量属性在饮片-物质基准间稳定传递.建立的栝楼桂枝汤指纹图谱和含量测定方法准确、可靠,可为栝楼桂枝制剂的质量控制提供技术参考.
HPLC fingerprint of substance benchmarks in Gualou Guizhi decoction and quantity transfer rule of key quality attributes
Objective:To establish a fingerprint of Gualou Guizhi decoction and a method for determination of multi-component to clarify the transfer rule of quantities and quality from decoction pieces and substance bench-marks.Methods:Fifteen batches of Gualou Guizhi decoction substance benchmarks were prepared and analyzed by the method of HPLC.Traditional Chinese medicine(TCM)Chromatographic Fingerprint Similarity Evaluation Software(2012)and hierarchical cluster analysis(HCA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)was used to evaluate the quality differences between batches of substance benchmarks,and screen the main chemical compositions that were led to quality discrepan-cy.The main differential components were determined by HPLC.And their contents,yields and transfer rates were analyzed for transfer rule of quantities and quality.Results:HPLC fingerprint of Gualou Guizhi decoction was established.A total of 30 common peaks in the fingerprint of Gualou Guizhi decoction substance benchmarks were assigned.Compared with the reference standards,9 components were identified as gallic acid,albiflorin,paeoni-florin,liquiritin,ononin,liquiritigenin,cinnamic acid,isoliquiritigenin and glycyrrhizic acid.The similarities of 15 batches of Gualou Guizhi decoction substance benchmarks samples were all above 0.920.All of them were divided into two categories by three chemical recognition modes for 5 major differential components,including gly-cyrrhizic acid,liquiritigenin,paeoniflorin,iquiritin,and ononin.The HPLC was also applied to determine the contents of multi-components and the method validation results were good.The average transfer rates of five index components were 51.65％,40.46％,74.74％,60.06％,and 34.54％,respectively and their average extraction rate was 14.20％.Conclusion:The critical quality properties of Gualou Guizhi decoction can be stably transferred from decoction pieces to substance benchmarks.The method of fingerprint and content determination is accurate and reliable,which is able to provide technique for quality control of Gualou Guizhi decoction formulation.
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