摘要
目的:建立聚合酶链式反应法(PCR法)快速鉴别川贝母真伪。方法:分析川贝母内转录间隔区1(ITS1)序列的特点,比较川贝母及其常见伪品的序列差异,设计了1对川贝母特异性扩增引物(CBM-SP引物),并用该引物对川贝母真品、伪品以及市场上购买的川贝母样品进行了鉴别。同时用《中华人民共和国药典》2015年版收录的聚合酶链式反应-限制性片段长度多态性法(PCR-RFLP)进行了验证。结果:用CBM-SP引物进行PCR鉴别,正品川贝母扩增出1条200 bp左右的条带,伪品未出现扩增条带。将该法应用于9批贝母商品药材的鉴定,可将川贝母及其伪品区别开来。本法实验结果与药典收录的PCRRFLP法实验结果一致。结论:本文建立的川贝母PCR快速鉴别法适用于川贝母及其常见伪品的真伪鉴别。
Objective:To establish a rapid polymerase chain reaction(PCR)method for the authentication of Fritillariae Cirrhosae Bulbus. Methods:The characteristics of the internal transcribed spacer 1(ITS1)of Fritillariae Cirrhosae Bulbus were analyzed. According to the sequence differences between Fritillariae Cirrhosae Bulbus and its common fake products,a pair of specific amplification primers(CBM-SP primers)were designed and used to identify the authentic Fritillariae Cirrhosae Bulbus,the fake products and the commercial Fritillariae Cirrhosae Bulbus samples. And the results were verified by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)method adopted in Chinese Pharmacopoeia(2015 edition). Results:The CBMSP primer was used for PCR identification. A band around 200 bp was detected in Fritillariae Cirrhosae Bulbus,while nothing appeared in other fake products. The method was applied in the identification of 9 batches of commercial samples of Fritillariae Cirrhosae Bulbus. Results showed that Fritillariae Cirrhosae Bulbus and its fake products could be distinguished. The above experimental results were consistent with the results of the PCRRFLP method adopted in the pharmacopoeia. Conclusion:The proposed rapid identification method of Fritillariae Cirrhosae Bulbus is applicable to the authentication of Fritillariae Cirrhosae Bulbus and its common fake products.
作者
刘香香
李靖
张煜彬
彭高发
兰燕宇
李勇军
王永林
杨畅
刘亭
LIU Xiang-xiang;LI Jing;ZHANG Yu-bin;PENG Gao-fa;LAN Yan-yu;LI Yong-jun;WANG Yong-lin;YANG Chang;LIU Ting(123451.Guizhou Provincial Key Laboratory of Pharmaceutics/State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Medical University,Guiyang 550004,China;School of Pharmacy,Guizhou Medical University,Guiyang 550004,China;Engineering Research Center for the Development and Application of Ethnic Medicine and TCM(Ministry of Education),Guizhou Medical University,Guiyang 550004,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2019年第10期1844-1851,共8页
Chinese Journal of Pharmaceutical Analysis
基金
国家自然科学基金项目(81603189)
关键词
聚合酶链式反应(PCR)
DNA分子鉴别
内转录间隔区(ITS)
川贝母鉴定
特异性引物
polymerase chain reaction(PCR)
DNA molecular authentication
internal transcribed spacer(ITS)
identification of Fritillariae Cirrhosae Bulbus
specific primers
