对市场上一种新的松贝伪品的比较研究  

Comparative study on a new origin of Songbei adulterant appeared in the market

作　　者：张文娟[1] 魏锋[1] 马双成[1] ZHANG Wen-juan;WEI Feng;MA Shuang-cheng(National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《药物分析杂志》2020年第4期659-663,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：国家科技部“重大新药创制”重大专项(2014ZX09304307-001,2014ZX09304307-002)。

摘　　要：目的:比较松贝与新出现伪品的不同特征,为鉴别检验提供依据。方法:综合性状、聚合酶链式反应-限制性内切酶多样性(PCR-RFLP)法、DNA序列比对、系统进化树以及遗传距离等方法,寻找松贝与新伪品的鉴别要点。结果:该贝母在性状上与松贝正品还是存在一定差异。按照《中华人民共和国药典》2015年版PCR-RFLP法检验,松贝PCR产物可以被完全酶切,而该伪品的PCR产物只能部分被酶切;测序分析可知,其部分酶切的原因是SmaⅠ酶切位点“CCCGGG”中第2位为C/T多态性位点。在内部转录间隔区(ITS区)266 bp范围内,该伪品与松贝比较有8个突变位点。系统进化树分析,该伪品与川贝母6个基原物种没有聚为一支。遗传距离分析,该伪品与川贝母6个基原物种的距离明显大于6个基原物种之间的距离。结论:该贝母的基原植物与松贝不同,应与松贝严格区分,以维护消费者经济利益,并为安全用药提供保障。Objective:To carry out comparative study on Songbei and its new adulterant,and to provide evidence for identification.Methods:Multiple analytical methods[appearance characteristics,polymerase chain reactionrestriction fragment length polymorphisms(PCR-RFLP),DNA sequence alignment,phylogenic analysis and heritage distance analysis]were utilized to identify Songbei and its new adulterant.Results:Some subtle difference in appearance characteristics could be found by carefully observation.The PCR product of the adulterant could be partially cutted according to the PCR-RFLP method in Pharmacopoeia of the People’s Republic of China(Chinese Pharmacopoeia)2015,while the PCR product of Songbei could be completely cutted.By sequencing analysis,a“C/T”polymorphism was found on the second position of SmaⅠrestriction site“CCCGGG”,which might be the reason why the PCR product was only partially cutted.There were 8 mutation sites within the length of 266 bp on the internal transcribed spacer(ITS)region of the adulterant,compared to that of Songbei.Phylogenic analysis of neighborjoining(N-J)tree showed that the adulterant failed to be clustered as one branch with the 6 kinds of originating plants of Fritillariae Cirrhosae Bulbus.The heritage distance analysis revealed a much greater distance between the adulterant and the 6 kinds of originating plants of Fritillariae Cirrhosae Bulbus,compared to the distance within the 6 kinds of originating plants.Conclusion:The origin of the adulterant is verified to be different from that of Songbei and should be strictly identified to assure the interest of consumers and the safety medicinal treatment.

关 键 词：川贝母 松贝 伪品 内部转录间隔区(ITS区) 聚合酶链式反应-限制性内切酶多样性(PCRRFLP) DNA序列比对 系统进化树 遗传距离 

分 类 号：R917[医药卫生—药物分析学]