经典名方一贯煎HPLC指纹图谱及化学模式识别研究

Study on HPLC fingerprint and chemical pattern recognition of the classical famous prescription Yiguanjian

目的:通过制备15批一贯煎基准样品冻干粉,建立一贯煎基准样品HPLC指纹图谱,为该经典名方的质量控制及评价提供参考。方法:采用Venusil C_(18)色谱柱(250 mm×4.6 mm,5μm),以甲醇(A)-0.1%磷酸水(B)为流动相梯度洗脱,建立15批一贯煎基准样品的HPLC指纹图谱,应用“中药色谱指纹图谱相似度评价系统”软件(2012版)进行相似度分析,并对共有峰进行归属。采用聚类分析(HCA),主成分分析(PCA)及正交偏最小二乘法-判别分析(OPLS-DA)对指纹图谱数据进行评价。结果:15批一贯煎基准样品与对照指纹图谱的相似度均≥0.771,共标定了31个共有峰,指认出了色谱峰11、20、23、25、26、30、31,分别为香草酸、阿魏酸、毛蕊花糖苷、异类叶升麻苷、补骨脂素、欧前胡素和蒿本内酯共7个成分;其中峰2、5~8、10、13、16~19、24、27归属于枸杞子,峰1、20、30、31归属于当归,峰21~23、25、28归属于生地黄,峰3归属于枸杞子和麦冬,峰4、29归属于枸杞子和生地黄,峰9归属于当归和沙参,峰11、12、14、15、26归属于枸杞子和当归,HCA、PCA及OPLS-DA可将15批基准样品大致分为2类。结论:建立的一贯煎HPLC指纹图谱方法专属性强,分离度好,灵敏度高,除川楝子药味以外均有表征,可为该方剂复方制剂的质量控制与评价提供参考。

Objective:To establish the Yiguanjian substance benchmark HPLC fingerprint by preparing 15 batches of Yiguanjian material benchmark freeze-dried powder,and provide a reference for the quality control and evaluation of this classic recipe.Methods:Using a Venusil C_(18) column(250 mm×4.6 mm,5μm)andmethanol(A)-0.1%phosphoric acid water(B)as mobile phase gradient elution,15 batches of Yiguan decoction material benchmark HPLC fingerprints were established.And the"Chinese Medicine"chromatographic fingerprint similarity evaluation system"software(2012 version)was performed to analyze similarity and assign common peaks.Cluster analysis(HCA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to evaluate the fingerprint data.Results:The similarity between the 15 batches of Yiguanjian material benchmark and the control fingerprint was≥0.771.A total of 31 common peaks were calibrated,and the chromatographic peaks 11,20,23,25,26,30,and 31 were identified,which were vanilla.Acid,ferulic acid,verbascoside,xenocimicifugaoside,psoralen,imperatorin,and artemisinol,including 7 components.Peaks 2,5-8,10,13,16-19,24 and 27 were attributed to Lycium barbarum L.,peaks 1,20,30,and 31 were attributable to Angelica sinensis(Oliv.)Diels,peaks 21-23,25,and 28 belonged to Rehmannia glutinosa Libosch.,peak 3 was attributable to Lycium barbarum L.and Ophiopogon japonicus(L.f)Ker-Gawl.,peaks 4 and 29 were attributable to Lycium barbarum L.and Rehmannia glutinosa Libosch.,peak 9 was attributed to Angelica sinensis(Oliv.)Diels and Glehnia littoralis Fr.Schmindt ex Miq.,and peaks 11,12,14,15,and 26 were attributed to Lycium barbarum L.and Angelica sinensis(Oliv.)Diels.HCA,PCA and OPLS-DA can roughly divide the 15 batches of substance standards into two categories.Conclusion:The established HPLC fingerprint method of Yiguanjian has strong specificity,good resolution,and high sensitivity.It has characteristics except for the medicinal taste of Melia toosendan Sieb.et Zucc.,which can provide a reference for the quality control and evaluation of the compound preparation of this formula.