摘要
目的:建立鸡内金配方颗粒特异性聚合酶链式反应(PCR)鉴别方法,用于原料-标准汤剂-配方颗粒一致性鉴定。方法:采用试剂盒法提取鸡内金药材、标准汤剂及配方颗粒基因组DNA模板,基于文献引物及初始PCR体系,优化PCR扩增体系及反应条件。结果:筛选并确定了PCR扩增体系及反应条件;鸡内金原料-标准汤剂-配方颗粒均可扩增出约279 bp的目标条带,而伪品及空白均无干扰;同时该方法适用于最大煎煮时长8 h的标准汤剂,并对鸡内金饮片检出限为0.33 ng,鸡内金配方颗粒检出限为2.32 ng。结论:该研究建立的鸡内金配方颗粒特异性PCR鉴别方法专属性好,灵敏度高,易于推广,为其他中药配方颗粒的质量控制提供了参考依据。
Objective:To establish a specific polymerase chain reaction(PCR)method for the consistency identification of Galli Gigerii Endothelium Corneum material-standard decoction-formula granules.Methods:The genomic DNA template of Galli Gigerii Endothelium Corneum material,standard decoction,formula granules was extracted by kit.PCR amplification system and reaction conditions were optimized based on primers in the literature and initial PCR system.Results:The PCR amplification system and reaction conditions were selected and determined.The target bands of Galli Gigerii Endothelium Corneum materials-standard decoction-formula granules were amplified at about 279 bp,and the forgeries and blanks and not interfered with it.The method could detect the sequence of Galli Gigerii Endothelium Corneum in standard decoction for 8 hours.The detection limit of decoction pieces was 0.33 ng,and the detection limit of formula grranules was 2.32 ng.Conclusion:This specific PCR identification method of Galli Gigerii Endothelium Corneum Formula established in this study has good specificity,high sensitivity and easy popularization,and can also be used as a reference for the quality control of other Chinese medicinal granula.
作者
李国卫
罗宇琴
宋叶
罗怡靖
谭斯尹
孙冬梅
陈向东
LI Guo-wei;LUO Yu-qin;SONG Ye;LUO Yi-jing;TAN Si-yin;SUN Dong-mei;CHEN Xiang-dong(Guangdong Yifang Pharmaceutical Co.,Ltd./Guangdong Provincial Key Laboratory of TCM Formula Granule Enterprise,Foshan 528244,China;China Medical University,Nanjing 210009,China;Guangzhou University of Chinese Medicine,Guangzhou 510006,China)
出处
《中药材》
北大核心
2025年第1期52-57,共6页
Journal of Chinese Medicinal Materials
基金
2022年佛山市南海区重点领域科技攻关专项(南科[2023]20号-18)。
关键词
鸡内金
鸡内金配方颗粒
聚合酶链式反应
Galli Gigerii Endothelium Corneum
Galli Gigerii Endothelium Corneum formula granules
Polymerase chain reaction
作者简介
李国卫(1989-),男,硕士,主任中药师,研究方向:中药配方颗粒质量控制及标准研究,E-mail:452048107@qq.com;通讯作者:孙冬梅,E-mail:tcmgdp@163.com。
