Study On Detection Method Of Bulbus Fritillariae And Pinellia Ternata In Huoxiangzhengqi Water Based On ARMS-PCR

The authenticity of Chinese medicinal materials and the quality of the Chinese patent medicines threaten human health.Unfortunately,the traditional identification methods have potential disadvantages.In recent years,gene-based tests have been widely used in identification of traditional Chinese medicine(TCM).Here,based on polymerase chain reaction(PCR),we study on identification of Bulbus Fritillariae and quantitative detection for Pinellia ternata in Huoxiangzhengqi water.We establish two methods,one of the methds is a fast detection method of medicinal materials,and the other one is a quantitative detection method for medicinal materials in preparation of TCM.The main content of this study is described as follows:1.Based on amplification refractory mutation system(ARMS)analysis,we developed three authentication scheme for the detection of Fritillaria pallidiflora.Meanwhile,based on ARMS analysis,we developed two authentication scheme for the detection of "Chuan Beimu".Universal primers as internal control can significantly reduce the false-negative rate.We can identify Fritillaria pallidiflora quickly,esasily,and reliablely by duplex PCR.What's more,the above methods can also be developed into a fast and high-throughput detection method by using SYBR Green.The above methods are accurate,reliable and specific,those works lay the foundation for multiplex PCR.2.We establish a quantitative detection method for Pinellia ternata in Banxia decoction by PCR.We successfully identify Pinellia ternata medicinal materials from adulterants.These results proved that the molecular authentication method were reliability and practicality.Then,we focus on the quantitative detection method for Pinellia ternata in Banxia decoction by PCR.The result suggested that accurate quantitative analysis of Pinellia ternata medicinal materials can be achieved by real-time fluorescent PCR assay and agarose-gel-electrophoresis based method.Finally,we explored to apply the quantitative analysis method to medicinal materials in Huoxiangzhengqi water.