分子生物学方法鉴别川贝母及饮片的技术要点探析

闫海霞+++谭成+++杜小伟+++陈有根+++傅欣彤

[摘要]中药是我国的传统药物，在临床和日常保健中有着越来越广泛的应用。中药材和中药饮片的质量关系到临床治疗的安全有效，对我国中医药产业的快速健康发展也至关重要。中药材和饮片的真伪鉴别一直是行业关注的焦点之一，随着生命科学技术飞跃发展，多种分子生物学技术被逐步运用于药用动植物及中药材的基源鉴定，取得较好的效果。《中国药典》自2010年版起，均收载了中药材和饮片（如蕲蛇、乌梢蛇和川贝母）的分子生物学方法，是采用新技术、新方法控制中药质量的有益尝试。标准已执行4年，但兄弟单位普遍反映在检验过程中常遇到各种技术问题，无法解决。本文以川贝母的分子生物学鉴别为例，通过总结实验过程中需要注意的技术要点以及常见问题，旨在为即将或正在从事这项检验工作的同仁们提供经验以供参考，以便更高效顺利地完成实验。

[关键词]分子生物学；川贝母；聚合酶链式反应-限制性片段长度多态性

[中圖分类号] R917 [文献标识码] A [文章编号] 1674-4721（2017）02（c）-0092-04

Technical essentials for molecular biological method on identification of Fritillaria cirrhosa and its decoction pieces

YAN Hai-xia TAN Cheng DU Xiao-wei CHEN You-gen FU Xin-tong▲

Department of Traditional Chinese Materials，Beijing Institute for Drug Control（Beijing Center for Health Food and Cosmetic Control），Beijing 102206，China

[Abstract]Traditional Chinese Medicine is traditional medicine of china，which has extensive use in clinic treatment and daily health-care.The quality of Chinese medicinal materials and Traditional Chinese Medicine decoction pieces relates greatly to the clinical curative effect and security，which is also very important to the development of Traditional Chinese Medicine property.The identification of Chinese medicinal materials and Chinese medicinal decoction pieces is a focal point in traditional Chinese medicines profession，with the development of life science and technology leap，a variety of molecular biology technology is gradually used in medicinal plants and animals and the base source identification of Chinese Medicinal Materials，achieved good results.The "Pharmacopoeia of the People′s Republic of China" since 2010，contains the Chinese medicinal materials and medicinal slices （e.g，Agkistrodon，wu pin snakes and Fritillaria cirrhosa） molecular biology method，is to adopt new technology，new method to control the quality of traditional Chinese medicine.Standard has been four years，but brother units generally reflect the various technical problems encountered in the inspection process，it can′t solve.Identification based on the molecular biology of Fritillaria cirrhosa，for example，by summing up the main points that need to pay attention to the process of technology and common problems，aiming at will or are engaged in the inspection work colleagues to provide experience for reference，so that more efficient to complete the experiment successfully.

[Key words]Molecular biology；Fritillaria cirrhosa；Polymerase chain reaction-restriction fragment length polymorphism中药作为中华民族的传统用药，在临床和日常保健中有着越来越广泛的应用。中药材和中药饮片的质量关系到临床疗效及用药安全，对我国中医药产业的快速健康发展至关重要[1]。中药材和中药饮片的真伪鉴别一直是行业关注的重点，随着科学技术的进步，分子生物学方法逐步被应用于中药材的鉴别[2-6]，2010年版《中国药典》及其第一增补本收载了蕲蛇、乌梢蛇和川贝母的分子生物学鉴别方法[7-8]，并在2015年版《中国药典》中得到修订和保留[9]。这是采用新技术新方法控制中药质量的有益尝试，但兄弟单位普遍反映在检验过程中常遇到各种技术问题，无法解决。北京市药品检验所2012年即开展蕲蛇和乌梢蛇的聚合酶链式反应（PCR）鉴别，以及川贝母的聚合酶链式反应-限制性内切酶长度多态性（PCR-RFLP）检验工作，至今已完成检品近200件，过程中积累一些经验和教训。笔者以川贝母为例，将检验过程中遇到的问题作一总结归纳。