目的 建立远志薄层色谱(thin-layer chromatography，TLC)及超高效液相色谱(ultra-high performance liquid chromatogra-phy，UHPLC)特征图谱，并分析远志全根、根皮和木心间化学成分差异，为远志质量评价提供参考。方法 以3，6'-二芥子酰基蔗糖、细叶远志苷A和远志皂苷B为指标成分建立TLC图谱;用UHPLC法建立远志特征图谱，结合热图聚类分析、主成分分析(principal component analysis，PCA)和正交偏最小二乘法判别分析(partial least squares discriminant analysis，OPLS-DA)评价多批远志全根、根皮和木心之间的差异性。结果 TLC图谱斑点清晰、分离度较好;UHPLC特征图谱共标定28个共有峰，指认了其中的7个特征峰，13批次远志特征图谱与对照图谱相似度均大于0。9，批次间质量较为稳定;TLC和UHPLC特征图谱显示远志全根、根皮和木心化学组成一致，未发现特异成分，热图聚类分析和PCA显示远志全根与根皮中各成分峰面积接近，OPLS-DA筛选出10个变量投影重要性(variable important in projection，VIP)值大于1的差异性成分。结论 所建立的TLC及UHPLC特征图谱简便、准确、稳定，展开系统和提取溶剂均使用低毒试剂，符合绿色化学理念，适用于远志的质量评价，从化学分析角度说明木心的存在对远志整体化学成分影响较小。
Study on thin-layer and ultra-high performance liquid characteristic chromato-grams of Polygalae radix
Objective To establish thin layer chromatography(TLC)and ultra-high performance liquid chromatography(UHPLC)characteristic chromatograms of Polygalae radix,to analyze the differences in chemical constituents among roots,cortexes and heartwoods,and to provide reference for the quality evaluation of Polygalae radix.Methods TLC characteristic chromatogram was established by using 3,6'-disinapoyl sucrose,tenuifoliside A and onjisaponin B as reference standards.UHPLC was used to estab-lish the characteristic chromatogram of Polygalae radix,and combined with heat map cluster analysis,principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)to evaluate the differences among roots,cortexes and heartwoods from various batches of Polygalae radix.Results The spots in TLC chromatogram were clear and well-separated.A total of 28 common peaks were demarcated from the characteristic chromatograms of Polygalae radix,and 7 characteristic peaks of which were identified.The similarities between the characteristic chromatograms of 13 batches of Polygalae radix and the reference chromatogram were all more than 0.9,indicating more stable in quality.Both TLC and UHPLC characteristic chromatograms showed that the chemical constituents of roots,cortexes and heartwoods of Polygalae radix were consistent,without any specific constituent.Heat map cluster analysis and PCA showed that the roots of Polygalae radix were similar in the peak area of each com-ponent to the cortexes,and OPLS-DA screened out 10 differential components via variable important in projection(VIP)values more than 1.Conclusion The established TLC and UHPLC characteristic chromatograms are simple,accurate and stable.The de-velopment system and extraction solvents used are low toxic,consistent with the concept of green chemistry.The methods are suitable for the quality evaluation of Polygalae radix.The chemical analysis indicates that heartwoods have few effects on the overall chemical composition of Polygalae radix.
Polygalae radixTLCUHPLCcharacteristic chromatogramquality evaluationremoval of heartwoodschemical pa-ttem recognition
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