蒙药短尾铁线莲多维鉴定研究  

Study on multidimensional identification of Clematis brevicaudata

作　　者：额尔敦呼 包同力嘎[2] 白明君 乌日汗[1] 苏门 乌日汉[1] 青白 包桂花[1] EERDUN Hu;BAO Tongliga;BAI Mingjun;WURI Han;Su Men;WURI Han;Qing bai;BAO Guihua(Mongolian Medical College,Inner Mongolia Minzu University,Inner Mongolia,Tongliao 028000,China;Clinical Mongolian Pharmacy Department of Affiliated Hospital of Inner Mongolia Minzu University,Inner Mongolia,Tongliao 028000,China)

机构地区：[1]内蒙古民族大学蒙医药学院,内蒙古通辽028000 [2]内蒙古民族大学附属医院临床蒙药剂部,内蒙古通辽028000

出　　处：《中国医药科学》2025年第7期64-68,共5页China Medicine And Pharmacy

基　　金：内蒙古自治区蒙医药标准化项目(2023-MB030);内蒙古自治区自然科学基金项目(2023LHMS08047)。

摘　　要：目的通过建立短尾铁线莲多维鉴定方法,为其制订质量标准提供依据。方法采用DNA条形码分子鉴定技术,对药材进行总DNA提取、ITS2序列聚合酶链式反应(PCR)扩增、双端测序、用DNAMAN、CodonCode ALigner 17.0软件以及ITS2 Database数据库构建碱基、峰图以及二级结构图;应用显微鉴定技术,对药材粉末进行过筛、制片、在生物显微镜下观察组织细胞特征;利用薄层色谱(TLC)鉴定技术,提取齐墩果酸和槲皮素、上行展开、加热、显色并检视。结果DNA条形码分子鉴定中,ITS2序列长度为299 bp、种内无变异位点,种内遗传距离为0,鸟嘌呤和胞嘧啶(GC)含量为65.55%,建立其碱基和峰图以及二级结构图;显微鉴定中分别鉴定出了叶上下表皮细胞、花粉粒、导管、纤维、非腺毛、气孔;薄层色谱鉴定中,分别鉴定出了齐墩果酸和槲皮素。结论研究表明ITS2基因序列可以鉴定短尾铁线莲,结合薄层色谱和显微鉴定法,均对短尾铁线莲进行准确鉴定,为制订质量标准体系提供科学依据。Objective To provide a basis for the development of quality standards for Clematis brevicaudata by establishing a multidimensional identification method for the herb.Methods Using DNA barcoding molecular identification technology,The total DNA was extracted,ITS2 sequence was amplified by polymerase chain reaction(PCR),double-ended sequencing was performed,and DNAMAN,CodonCode ALigner 17.0 software and ITS2 Database were used to construct the base,peak map and secondary structure map.The microscopical identification technique was used to screen and slice the medicinal powder,and the histological cell characteristics were observed under the biological microscope.Oleanolic acid and quercetin were extracted by thin layer chromatography(TLC),unfolded upward,heated,colored and examined.Results In the molecular identification of DNA barcode,the length of ITS2 sequence was 299 bp,no intra-species variable site,the intra-species genetic distance was 0,and the content of guanfacine and cytosine(GC)was 65.55%,and the establishment of its base and peak diagrams as well as secondary structure diagrams,in the microscopic identification,the upper and lower epidermal cells of the leaf,pollen grains,ducts,fibers,non-glandular hairs,and stomata were identified,respectively,in the TLC identification,respectively Oleanolic acid and quercetin were identified in thin-layer chromatography.Conclusion The ITS2 gene sequence can be used to identify the short-tailed clematis,and the combination of thin-layer chromatography and microscopic identification method can accurately identify the Clematis brevicaudata,which can provide a scientific basis for the development of quality standard system.

关 键 词：短尾铁线莲 脱氧核糖核酸条形码 薄层色谱法 显微鉴别法 

分 类 号：R248.1[医药卫生—中医临床基础]