Separation And Preparation Of Chemical Constituents Of Cassia Seeds And Study On Comparative Extracts

This dissertation includes three parts.The first part:summary of the related literaturesThis chapter reviews the development status,advantages and application of the Chinese medicine standard extracts.At the same time,this chapter also reviews chemical composition,pharmacological effects and quality evaluation method of cassia,and provide ideas for further improvement of quality control system of cassia.The second part:Study on the chemical constituents of cassiaThe HPLC-IT-TOF MS method is used to study the chemical constituents of cassia identification.There are 17 compounds inferred,including 7 naphthopyrones,7 anthraquinones and 3 naphthoquinones.Among in that,the research group selects the three naphthopyrones(cassiaside B2,rubrofusarin-6-O-?-D-gentiobioside and cassiaside C)as the main components of the standard extract.The research group uses a variety of isolation and structural identification technique to research chemical constituents of cassia.Five compounds are isolated and identified,including 2 anthraquinones(aurantiobtusin-6-O-?-D-glucopyranoside and chrysophanol-1-O-?-D-glucopyransyl-(1-3)-O-?-D-glucopyransyl-(1-6)-O-?-D-glucopyranoside)and 3 naphthopyrones(cassiaside B2,rubrofusarin-6-O-?-D-gentiobioside and cassiaside C).Three naphthopyrones are calculated by peak area normalization method.It is showed that the purity of the three compounds is more than 98%.So the three compounds can be used as a chemical reference to research cassia standard extract.The third part:the research and application of the standard extractfrom cassiaAn HPLC method is established for determinating of three naphthopyrones including cassiaside B2,rubrofusarin gentiobioside and cassiaside C simultaneously in cassia.The results show that the method is simple,well repeatable and reproducible.The research group studies extraction process and preparation method of cassia standard extract,with cassiaside B2,rubrofusarin-6-O-?-D-gentiobioside and cassiaside C as indexes.The prepared route of cassia standard extract is established for the first time.After the purified preparation,the total content of three main components reaches above 60%.The preparation method is stable,reproducible,and provides research base for the development of Chinese medicine standard extracts.The research group uses qualitative analysis,quantitative analysis,characteristic chromatogram and determination of moisture content to research cassia standard extract.(1)Qualitative analysis:the research group uses LC-MS method and TLC method to study cassia standard extract.The results show that the cassias tandard extract containes cassiaside B2,rubrofusarin-6-O-?-D-gentiobioside and cassiaside C.(2)Quantitative analysis:HPLC method is used to determine the three main components of cassia standard extract.The results show that the method is well repeatable and reproducible.(3)Characteristic chromatogram:the research group establishs characteristic chromatogram of ten batches of cassia standard extract.The results show that the similarity of ten batches of cassia standard extract is above 0.9.(4)Determination of moisture content:the research group uses Karl Fischer coulometry to determine the water content of cassia standard extract.The average moisture content is 2.12%,which achieve relevant standard.The homogeneity and stability of cassia standard extract are tested.The cassia standard extract showes good uniformity and stability.HPLC method is used to determine the three main components and to study certification as well as uncertainty of cassia standard extract.The results of certification and uncertainty of three main components are(19.92±0.86)%,(21.49±0.88)%,(24.08±0.98)%.The uncertainties of the three components are all less than 1%.This suggests that uncertainty is normal and reliable.The cassia standard extract has good reproducibility between different laboratories and different instruments,which shows the feasibility of promotion of cassia standard extract.The research group establishes the qualitative identification method of cassia with standard extractas index,which make an important progress and development to the cassia qualitative identification.The research group establishes the content determination method of cassia with standard extract as index.Comparing the determination results of the method using standard extractas index and the method using reference substance as index,it shows no significant difference between the results of two quantitative methods.The result illustrates that standard extract can replace reference substance in cassia quality control.The research group explores quality control standards of cassia based on the content determination method using standard extract as index.Three limits of quality evaluation for cassia are initially established,which provide some suggestions for the improvement of cassia quality evaluation system.