滇西北野生滇龙胆根部HPLC指纹图谱研究

Study on HPLC Fingerprints of Roots of Gentiana rigescens Franch. in Northwestern Yunnan

目的建立滇西北少数民族聚居地区野生滇龙胆根部的高效液相色谱(HPLC)指纹图谱,为其质量评价提供依据。方法采用HPLC法,色谱柱为Agilent Intersil-C18(4.6 mm×250 mm,5μm),流动相为0.1%甲酸水溶液(A)和乙腈(B),梯度洗脱,柱温30℃,流速1 ml·min-1,检测波长241 nm,进样量5μl。建立10批滇龙胆根部指纹图谱,采用"中药色谱指纹图谱相似度评价系统(2004A)"确定共有峰,计算共有峰相对保留时间和相对峰面积,进行相似度分析,并采用SPSS22.0软件进行聚类分析。结果滇龙胆根部指纹图谱共确定17个共有峰,图谱相似度大于0.9,10批样品聚为两类,发现各批次样品成分类型基本一致,成分含量存在差异。检测出5种主要成分马钱苷酸、獐芽菜苷、龙胆苦苷、当药苷和异牡荆苷的含量,各批次样品龙胆苦苷含量均大于3.0%。结论该方法稳定、可靠,可为滇西北滇龙胆药材质量评价及少数民族聚居地区药用植物资源开发提供依据。

Objective To establish HPLC fingerprints of roots of Gentiana rigescens Franch. in ethnic minority areas of Northwestern Yunnan and provide the basis for quality evaluation of this species. Methods HPLC method was used on Agilent Intersil-C18column（ 4. 6 mm × 250 mm,5 μm） with the mobile phase of 0. 1% formic acid water（ A） and acetonitrile（ B） for gradient elution. The column temperature was 30 °C and the flow rate was 1. 0 m L/min. The detection wavelength was set at 241 nm,and the injection volume was 5 μl. The HPLC fingerprints for roots of G. rigescens from ten different origins were established. At the same time,the similarity evaluation system for chromatographic fingerprint of TCM（ version 2004A） was used for identifying the common peaks,calculating relative retention times and relative peak areas,as well as performing similarity analysis. Besides,cluster analysis was carried out by using SPSS 22. 0 software. Results 17 common peaks of fingerprints of roots of G. rigescens were identified,and the similarities were greater than 0. 9. Ten batches of samples were classified into two groups,and it showed that these samples have similar constituents and the contents of these constituents were different. The contents of four main components of loganic acid,swertiamarin,gentiopicroside,sweroside and isovitexin were tested. The content of gentiopicroside in each batches of samples was greater than 3. 0%. Conclusion This method is stable and reliable and it can be used for quality evaluation of G. rigescens in the northwest of Yunnan Province. What’s more,it could provide reliable basis for the development of medicinal plant resources in ethnic minority areas.