深绿卷柏的显微鉴别和高效液相色谱特征图谱研究  

Microscopic identification and HPLC characteristic chromatogram of Selaginella doederleinii Hieron

作　　者：王新桂 朱田密[1,2] WANG Xin-gui;ZHU Tian-mi(Hubei Hospital of Traditional Chinese Medicine/Affiliated Hospital of Hubei University of Chinese Medicine,Wuhan 430061;Hubei Academy of Traditional Chinese Medicine,Wuhan 430074)

机构地区：[1]湖北省中医院湖北中医药大学附属医院,武汉430061 [2]湖北省中医药研究院,武汉430074

出　　处：《中南药学》2023年第3期758-763,共6页Central South Pharmacy

基　　金：全国中药特色技术传承人才培训项目[国中医药人教函(2019)43号]。

摘　　要：目的研究深绿卷柏的显微鉴别特征,并建立其HPLC特征图谱。方法采用茎横切片法和叶表面制片方法研究深绿卷柏的显微特征。色谱柱为Agilent ZORBAX SB-C18 Narraw-Bore(2.1 mm×150 mm,5μm),以乙腈-0.1%磷酸水为流动相进行梯度洗脱,流速为0.25 mL·min^(-1),检测波长为330 nm,柱温为30℃,记录15批深绿卷柏的色谱图,计算相似度,确认10个共有峰为特征峰,以穗花杉双黄酮色谱峰为参照,计算相对保留时间,建立特征图谱,对部分特征峰进行指认,且与3种混淆品特征图谱进行比较。用SPSS软件对深绿卷柏进行聚类分析。结果深绿卷柏的茎、叶显微特征明显。15批深绿卷柏样品的相似度为0.765~0.964,特征峰稳定,并且可以与混淆品中华卷柏、江南卷柏、卷柏相区别。指认了4种双黄酮类成分,分别为穗花杉双黄酮、罗波斯塔双黄酮、橡胶树双黄酮、穗花杉双黄酮-7,4',7'',4'''-四甲醚。聚类分析将深绿卷柏分为两类,与产地相关。结论所建立的方法简便、实用,可为深绿卷柏的鉴别提供依据。Objective To determine the microscopic characteristics and identify the characteristic chromatogram method of Selaginella doederleinii Hieron.by HPLC.Methods Stem transverse sectioning and leaf sealing were used in the microscopic study.Agilent ZORBAX SB-C18 Narraw-Bore(2.1 mm×150 mm,5μm)column was used with gradient elution with acetonitrile-0.1%phosphoric acid solution as the mobile phase at a flow rate of 0.25 mL·min^(-1).The detection wavelength was 330 nm,the column temperature was 30℃.The HPLC chromatogram of 15 batches of Selaginella doederleinii Hieron.samples was recorded and similarity calculated.Totally 10 common peaks were confirmed as characteristic peaks.Each relative retention time was calcutated with Amentoflavone peak as the reference.The characteristic chromatogram was estabilshed and compared with 3 confusing varieties.Some characteristic peaks were identified.Hierarchical cluster analysis was performed with SPSS.Results The microscopic characteristics of the stem and leaves were obvious.The similarity of 15 batches of Selaginella doederleinii Hieron.ranged from 0.765 to 0.964.The characteristic peaks were stable and could be distingushed from adulterants such as Selaginella sinensis,Selaginella moellendorffii and Selaginella tamariscina.Four biflavonoids in the characteristic chromatogram of Selaginella doederleinii Hieron.were identified as amentoflavone,robustaflavone,heveaflavone,and 7,4',7'',4'''-tetra-O-methyl-amentoflavone.The 15 samples from different areas were divided into 2 clusters by hierarchical cluster analysis. Conclusion The method is simple and practical, which can be used to identify authentic Selaginella doederleinii Hieron..

关 键 词：深绿卷柏 显微鉴别 特征图谱 高效液相色谱 

分 类 号：R284.1[医药卫生—中药学]