青天葵高效薄层色谱指纹图谱的鉴别研究

Identification of High-performance Thin-layer Chromatographic Fingerprints for Nervilia fordii(Hance) Schltr.

目的建立青天葵药材指纹图谱质量控制方法。方法采用Merck硅胶60高效预制薄层板,氯仿∶乙酸乙酯∶甲醇∶甲酸(4.5∶1.5∶2.5∶0.4)为展开剂,10%硫酸乙醇溶液显色,366 nm波长下扫描吸收光谱,测定18批青天葵药材的薄层色谱图谱,采用指纹图谱系统解决方案软件进行指纹图谱分析,与随行黄酮对照品比较,辨认色谱峰。结果建立青天葵药材薄层色谱指纹图谱,确定7个共有峰;与随行对照品比较,辨认了其中的5个色谱峰分别为鼠李秦素-3-O-β-D-葡萄糖苷、青天葵甲素、青天葵乙素、沙苑子苷和青天葵丙素,均为黄酮类成分;对18批次青天葵药材指纹图谱主成分分析,根据主成分得分,筛选出12批次合格样品导入指纹图谱系统解决方案软件,建立及生成共有模式,并进行相似度评价,结果 7批大叶青天葵和5批中叶青天葵指纹图谱与共有模式相似度>0.90,3批小叶青天葵及5批毛叶青天葵相似度均低于0.75;紫背天葵和天葵的薄层色谱与青天葵在相同位置无相同的荧光斑点。结论所建立的青天葵黄酮指纹图谱具有专属性强、操作简便、快速的特点,可有效地鉴别不同商品规格的青天葵药材、青天葵代用品"毛叶青天葵"以及"同名异物"紫背天葵和天葵,可用于青天葵药材的鉴别和质量评价。

Objective To establish the fingerprint profiling of Nervilia fordii（Hance）Schltr.（Qingtiankui）for its quality control. Methods High-performance thin-layer chromatography（HPTLC）was adopted. The separation was carried out on HPTLC pre-coated plates（silica gel Merck 60）using mixture of CHCl3（chloroform）∶Et OAc（ethyl acetate）∶Me OH（methanol）∶HCOOH（formic acid）at the proportion of 4.5 ∶1.5 ∶2.5 ∶0.4 as the mobile phase. Visualization of the chromatogram was performed by spraying with 10 % sulphuric acid-ethanol solution and heating at 105 ℃ until the bands were visible. The plate was observed at UV 366 nm and the HPTLC fluorescence image was documented. The corresponding digital scanning profile was generated and evaluated by software of Computer-assisted Fingerprints Evaluation System. Eighteen patches of Qingtiankui were determined and evaluated by similarity analysis and principal component analysis（PCA） with flavonoids as reference. Results The common pattern of Qingtiankui was established and 7 common peaks were confirmed. With comparison of the reference, 5 common peaks were identified as rhamnazin-3-O-β-D-glucoside, nervilifordizin A, nervilifordizin B, complanatuoside and nervilifordizin C.According to the scores of PCA,12 batches of qualified samples were screened out and their digital scanning profile was evaluated by CHROMAP 1.5 Software. The results of similarity analysis showed that the similarity of 7 batches of large-leave Qingtiankui and 5 batches of moderate-leave Qingtiankui was over 0.90,and the similarity of 3 batches of small-leave Qingtiankui and 5 batches of hairy-leave Qingtiankui was less than 0.75. No same fluorescence spot was found at the same positon of thin-layer chromatograms of Begonia fimbristipula Hance（Zibeitiankui）and Semiaquilegia adoxoides（DC.）Makino（Tiankui）. Conclusion This method is simple,repeatable and specific,and it can be used to control the quality of Nervilia fordii（Hance）Schltr.（Qingtiankui）,and also can be used to identify its substitute hairy-leave Qingtiankui or its adulterants Begonia fimbristipula Hance（Zibeitiankui） and Semiaquilegia adoxoides（DC.）Makino（Tiankui）.