分子生物學方法鑒別川貝母及飲片的技術要點探析

閆海霞+++譚成+++杜小偉+++陳有根+++傅欣彤

[摘要]中藥是我國的傳統藥物，在臨床和日常保健中有著越來越廣泛的應用。中藥材和中藥飲片的質量關系到臨床治療的安全有效，對我國中醫藥產業的快速健康發展也至關重要。中藥材和飲片的真偽鑒別一直是行業關注的焦點之一，隨著生命科學技術飛躍發展，多種分子生物學技術被逐步運用于藥用動植物及中藥材的基源鑒定，取得較好的效果。《中國藥典》自2010年版起，均收載了中藥材和飲片（如蘄蛇、烏梢蛇和川貝母）的分子生物學方法，是采用新技術、新方法控制中藥質量的有益嘗試。標準已執行4年，但兄弟單位普遍反映在檢驗過程中常遇到各種技術問題，無法解決。本文以川貝母的分子生物學鑒別為例，通過總結實驗過程中需要注意的技術要點以及常見問題，旨在為即將或正在從事這項檢驗工作的同仁們提供經驗以供參考，以便更高效順利地完成實驗。

[關鍵詞]分子生物學；川貝母；聚合酶鏈式反應-限制性片段長度多態性

[中圖分類號] R917 [文獻標識碼] A [文章編號] 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中藥作為中華民族的傳統用藥，在臨床和日常保健中有著越來越廣泛的應用。中藥材和中藥飲片的質量關系到臨床療效及用藥安全，對我國中醫藥產業的快速健康發展至關重要[1]。中藥材和中藥飲片的真偽鑒別一直是行業關注的重點，隨著科學技術的進步，分子生物學方法逐步被應用于中藥材的鑒別[2-6]，2010年版《中國藥典》及其第一增補本收載了蘄蛇、烏梢蛇和川貝母的分子生物學鑒別方法[7-8]，并在2015年版《中國藥典》中得到修訂和保留[9]。這是采用新技術新方法控制中藥質量的有益嘗試，但兄弟單位普遍反映在檢驗過程中常遇到各種技術問題，無法解決。北京市藥品檢驗所2012年即開展蘄蛇和烏梢蛇的聚合酶鏈式反應（PCR）鑒別，以及川貝母的聚合酶鏈式反應-限制性內切酶長度多態性（PCR-RFLP）檢驗工作，至今已完成檢品近200件，過程中積累一些經驗和教訓。筆者以川貝母為例，將檢驗過程中遇到的問題作一總結歸納。