基于DNA指纹图谱的水牛角真伪及掺杂鉴别方法研究  

Authenticity and adulteration identification of Bubali Cornu based on DNA fingerprints

作　　者：熊辕 陈天韵 张恬[1] 田娜 蒋超[1] XIONG Yuan;CHEN Tian-yun;ZHANG Tian;TIAN Na;JIANG Chao(State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院、中药资源中心、道地药材品质保障与资源持续利用全国重点实验室,北京100700

出　　处：《中国中药杂志》2024年第7期1826-1833,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82073972);中国中医科学院优秀青年科技人才培养专项(ZZ13-YQ-089)。

摘　　要：是否存在掺杂是中药鉴定的难点问题。水牛角主要以水牛角丝或水牛角粉2种形态流通于药材市场,缺乏明显的鉴定特征,存在掺杂风险。然而目前水牛角的掺杂鉴定方法缺失。为解决中药水牛角的真伪及掺杂问题,控制药材水牛角的质量,保证临床用药的真实性。使用DNA指纹图谱对来自药材企业和药材市场的43批样品进行鉴定,筛选出可鉴定水牛角和牦牛角的荧光DNA指纹图谱扩增引物,利用荧光毛细管分型技术获得水牛角的DNA指纹图谱。并检测了荧光毛细管分型技术对不同掺杂比例的鉴别效果。筛选出了2对可区分水牛角和牦牛角的荧光STR分型引物16Sa和CRc,建立DNA指纹图谱鉴别方法,牦牛角16Sa迁移峰为223.4~223.9 bp、水牛角为225.5~226.1 bp,牦牛角CRc迁移峰为518.8~524.8 bp、水牛角为535.9~542.5 bp。通过迁移峰的峰高可对掺伪比例在50%以下的掺杂品进行初步定量,定量结果与掺入比例相似。该研究建立了一种简单快捷,可用于中药水牛角及其掺杂品的通用DNA指纹图谱鉴别方法,能够实现对药材水牛角鉴定的同时对掺杂品进行半定量。Whether adulteration exists is a difficult problem in the identification of traditional Chinese medicine(TCM).Bubali Cornu is mainly available in the medicinal material market in the form of buffalo horn silk or buffalo horn powder but lacks obvious identification characteristics,so there is a risk of adulteration.However,the method of identification of adulteration in Bubali Cornu is lacking at present.In order to ensure authenticity and identify adulteration of TCM Bubali Cornu,control the quality of TCM Bubali Cornu,and ensure the authenticity of clinical use,the DNA fingerprints of 43 batches of samples from pharmaceutical companies and medicinal material markets were identified,and the amplification primers of fluorescent DNA fingerprints of Bubali Cornu and Bovis Grunniens Cornu were screened.The DNA fingerprints of Bubali Cornu were obtained by fluorescent capillary typing.The identification effect of fluorescent capillary typing on different adulteration ratios was also tested.Two pairs of fluorescent STR typing primers,namely 16Sa and CRc,which can distinguish Bubali Cornu and Bovis Grunniens Cornu,were screened out,and a DNA fingerprint identification method was established.The 16Sa migration peaks of Bovis Grunniens Cornu and Bubali Cornu were 223.4-223.9 bp and 225.5-226.1 bp.The CRc migration peaks of Bovis Grunniens Cornu and Bubali Cornu were 518.8-524.8 bp and 535.9-542.5 bp.The peak height of the migration peak could be used for preliminary quantification of the adulterants with an adulteration ratio below 50%,and the quantitative results were similar to the adulteration ratio.In this study,a simple and quick universal DNA fingerprint method was established for the identification of Bubali Cornu and its adulterants,which could realize the identification of TCM Bubali Cornu and the semi-quantitative identification of the adulterants.

关 键 词：水牛角 掺杂 DNA指纹图谱 STR分型 分子鉴别 动物药 

分 类 号：R282.5[医药卫生—中药学]