摘要
目的改进《中国药典》2015版中川贝母分子鉴别方法,为《中国药典》的应用和标准修订提高提供参考。方法优化聚合酶链式反应(PCR)模板与引物的比率,提高退火温度至61℃;采用乙醇沉淀法纯化PCR反应液。结果原方法 PCR出现明显非特异性条带,酶切结果模糊。方法改进后,非特异性条带得到明显抑制,同时目的条带量没有明显影响,酶切结果清晰,效果良好。结论该方法专属性强、重现性好,可以满足川贝母的鉴别。
Objective To improve the molecular identification method of Fritillariae cirrhosae bulbus in Chinese Pharmacopoeia（ 2015version）,and to provide reference for implementation of the standards of Chinese Pharmacopoeia. Methods Primer quantity was reduced while template quantity was improved. And the annealing temperature was changed from 56 ℃ to 61 ℃. Ethanol precipitation was used for PCR purification. Results Using original method,one nonspecific band（ 200 bp to 300 bp） was found in PCR result and the result of restriction endonucleases digestion was too fuzzy to identify. After the method was improved,the nonspecific band was obviously inhibited,while the purpose band was not significantly affected. The result of restriction endonucleases digestion was clear for identification. Good results were achieved with the improved method. Conclusion This method has good repeatability and can be used for molecular identification of Fritillariae cirrhosae bulbus.
出处
《安徽医药》
CAS
2016年第10期1872-1875,共4页
Anhui Medical and Pharmaceutical Journal
关键词
川贝母
分子鉴别
中药质量标准
Fritillariae cirrhosae bulbus
Molecular identification
Quality standard of traditional Chinese medicine
