Key Technologies For Quality Evaluation Of Galli Gigerii Endothelium Corneum And Chinese Patent Medicines Containing Galli Gigerii Endothelium Corneum

Galli Gigerii Endothelium Corneum(GGEC),a common TCM for both medicine and food,is the dry stomach inner-wall of Gallus gallus domesticus(Chinese Pharmacopoeia Committee2020).GGEC is used for the treatment of vomiting,infantile food accumulation,diarrhea,enuresis,spermatorrhea,biliary depression and other diseases.It is recorded in every edition of Chinese Pharmacopoeia.Meanwhile,GGEC is also widely used as health food,and market demand is large.Therefore,there are a lot of substitution,adulteration,and aggravation phenomena,and even adulterated with some from sick chicken.The safety of medication was affected.The main components of GGEC are amino acids,proteins,polysaccharides and so on.It is often difficult to use the identification technology commonly used in TCM decoction methods.The identification of GGEC in Chinese Pharmacopoeia(2020 version)still relies on the traditional character identification,and lacks effective modern methods.In this study,a variety of modern technology were used to study the authentication of GGEC in substitution,adulteration,microbial safety,that provide a reference for the appraisal standard of GGEC and even animal origin medicine.The main parts of study are as follows:1.In this research,an electronic-nose method was established to deepen the accuracy of character identification.The traditional character identification is mainly based on the shape,color,cross-section,and smell of medicine.It is subjective and highly dependent on the experience of the appraisers to realize the identification by relying on the appraisers to look,smell and taste.In this experiment,the optimized electronic nose detection technology was used to quantify the volatile odor of GGEC,to determine the difference between GGEC powder and substitute,and to achieve the purpose of identification through cluster analysis.By optimizing the detection conditions,the electronic-nose accurately distinguished the authenticity of GGEC,and accurately distinguished GGEC and processed GGEC.The results of this session show that the electronic-nose technology can effectively improve the reliability of GGEC character identification.2.Multiplex PCR technology was established to identify the GGEC adulteration of substitutes.In the process of collecting GGEC,the substitutes from Anas platyrhynchos and Anser anser are often added intentionally or unintentionally.They have similar appearance and similar chemical composition.It is difficult to distinguish the adulteration by common spectral and chromatographic techniques.In this research,a multiple species-specific PCR method was established to identify the adulteration of GGEC and substitutes,and verified by self-control.In this study,we optimized the reaction system and reaction conditions of PCR,and investigated its sensitivity,specificity,and applicability.The established method has high specificity,and the detection limit is 1 ng/μL.Through the optimized technology,the substitutes from Anas platyrhynchos and Anser anser were identified from the artificially mixed samples,and 2 batches with substitute from Anser anser were identified from 14 batches of GGEC commodities.The results showed that the multiplex PCR technology developed in this chapter can be used to identify the adulteration of GGEC.3.The multiplex PCR was optimized to realize the authenticity identification of GGEC in Chinese patent medicine.Chinese patent medicine was made from ready-to-use materials in crushed or extracted forms,which cannot be identified by character.In this research,based on the above multiple species-specific PCR technology,by optimizing the reaction system and reaction conditions of PCR method,and investigating its sensitivity and specificity,the established detection method has high specificity,and the detection limit is 1 ng/μL.Through the optimized technology,2 batches of adulterants by Anas platyrhynchos and 1 batch of adulterants by Anser anser were identified from the commodity "Xiao Er Fufang Jineijin San" and "Xiao Er Feike Keli".The results show that the multiplex PCR technique developed in this chapter can be used to identify the adulteration of GGEC in Chinese patent medicine.4.The fluorescent quantitative PCR technology was established to identify the contamination in GGEC by Escherichia coli,which should not be detected in TCM in Chinese Pharmacopoeia.There are many kinds of microorganisms attached to TCM,but the amount of DNA extracted is very small,so it is difficult to detect by common PCR technology.In this study,the method of q PCR was designed and optimized,and E.coli was detected in 14 batches of commercial GGEC.Compared the detection results with the conventional PCR and plate culture count results,the method showed a high sensitivity and accuracy.