Quantitative Analysis Of Alkaloids And Nucleotides In Medicinal Herbs Of Fritillaria

Objective:Using modern analysis and combination technology to study the medicinal materials of Fritillaria,to investigate the distribution of nucleoside and alkaloids in different Fritillaria,and to compare the differences between species,and to provide an important guarantee for the identification and quality control of Fritillaria.To explore the advantages and disadvantages of different detectors in the analysis of alkaloids,and to provide new ideas and methods for the study of Fritillaria.Methods:1.Using HPLC-DAD method to determine the content of nucleoside in the medicinal materials of Fritillaria.2.The contents of alkaloids in Fritillaria medicinal herbs were determined by HPLC-ELSD method and HPLC-CAD method,and the differences between different detectors were compared.3.Using SPSS data analysis software to cluster analysis of quantitative results.Results:1.Through the exploration of HPLC-DAD method,the best experimental scheme was established,the chromatographic column was Thermo Hypersil Gold chromatographic column,methanol-water as mobile phase,gradient elution,sampling quantity 20μl,column temperature 25℃,Flow rate 1 mL.min^-1,dad detector detection wavelength:260nm.The recovery rate of 9 nucleoside compounds was between 95%¹05%and RSD was within 2%.The content of 9 nucleoside compositions was obtained,and 9 nucleoside compositions were distributed in different species of Fritillaria and the difference was not obvious.2.Through the investigation of extraction method,the method of alkaloid extraction was determined as ammonia infiltration alkali 2h,chloroform:Methanol（4:1）as extraction solvent,heating reflux 120min.In order to find the best experimental scheme,the chromatographic column is Thermo Hypersil Gold chromatographic column,the flow phase is 0.08%ethylamine-acetonitrile,gradient elution,sampling quantity 20μl,column temperature 35℃,flow rate 1 mL.min^-1.ELSD Detector Parameters:The carrier velocity is 2.5L.min^-1,the drift tube temperature is 105℃.The recovery rate of 5 alkaloids was between 95%¹05%and RSD was within 2%.The content of Peimisine,Sipeimine-3β-D-glucoside,Sipeimine,Peimine and Peiminine were obtained,and the differences between different Fritillaria were found by cluster analysis.3.Through the exploration of the HPLC-CAD method,the best experimental scheme is established,the chromatographic column is Zorbax sb-c18 chromatographic column,the flow phase is 0.1%trifluoroacetate-acetonitrile,gradient elution,the sampling quantity put 10μl,the column temperature 30℃,and the flow velocity 1mL.min^-1,fog detector parameters:Atomization Temperature 35℃,gas source for liquid nitrogen,pressure 4.206x105Pa.The recovery rate of 5 alkaloids was between95%¹05%andRSDwaswithin2%.ThecontentofPeimisine,Sipeimine-3β-D-glucoside,Sipeimine,Peimine and Peiminine were obtained and the content of Fritillaria was analyzed by cluster analysis in order to identify the different medicinal herbs.4.Different detectors（ELSD detectors and CAD detectors）of the same 5 alkaloid content determination results are basically consistent,however,as a new type of CAD detector,it has the advantages of sensitivity,better stability,lower ration limit,and can detect the ELSD material or the material which could not be detected.Conclusion:Through the above research,the content determination results of the Hplc-dad method were analyzed,and the total contents of 9 nucleoside compositions,8 species of Fritillaria,were found,and the content was not significant in species,which might be the commonness and regularity of different fritillaria herbs,and the content determination result of HPLC-ELSD method was studied,The contents of 5alkaloids from 8 species of Fritillaria were obtained,and the results of cluster analysis and later analysis were used to identify the medicinal herbs of fritillaria.The contents of 5 alkaloids from 8 Fritillaria were determined by using a new type of CAD detector.More accurate and reliable distribution of 5 kinds of alkaloids in medicinal herbs,according to the results of cluster analysis,it is more clear to distinguish different fritillaria medicinal herbs,and it is found that CAD detector is more suitable for the quantitative analysis of alkaloids in Fritillaria,which provides a new thought and method for quality control of Fritillaria.It provides an important basis for clinical safe drug use,and through the combination of the modern analysis,it can comprehensively analyze the medicinal materials of fritillaria and provide reliable means for its quality control,and guarantee the quality of medicinal materials and the safety of clinical medication.