The Chemical Composition Analysis Of Sophora Flavescens And The Preparation Technology Of Sophoridine And 13,14-dehydrosophoridine

Sophora alopecuroides L.belongs to the Leguminous species,it flavor is bitter and property is cold,with a little toxicity.As a traditional Chinese herbal medicine,it has the function of clearing the intestines and dampness,and it be used for acute dysentery and enteritis.It is a traditional Chinese medicine in the Northwest northwestern region.Based on the study of the chemical constituents of S.alopecuroides,this paper studied the preparation process of sophoridine and 13,14-dehydrosophoridine.This paper is divided into 4 chapters.Part 1 Summary of the related literaturesThe clinical application,chemical composition,pharmacological action and preparation process of alkaloids of kudouzi(Sophora alopecuroides L.)were reviewed,which provided theoretical basis for the subsequent work.Part 2 Study on Chemical Constituents of S.alopecuroidesThe air-deried seeds S.alopecuroides were reflux extracted by ethanol.Solvent extraction and various chromatographic techniques(silica gel,polyamide,C18,Sephadex LH-20,etc.)were used to isolate the chemical constituent of extract.The structure of this compounds were identified by using physicalchemical prorerties and sepectralanlysis(ESI-MS,1H NMR,13C NMR,and HMBC,HMQC).Nine alkaloids were obtained and indetified,there are sophocarpine,Matrine,N-metylcytisine,sophoridine,cytisine,oxymatrine,oxysophocarpine,sophoramine and 13,14-dehydrosophoridine.One Chalcone is Butine 4-?-(3-D-glucop-yanoside.There are three others,there are ?-amyrin,daucosterol,and?-sitosterol.Among this constituents,?-amyrin and Bute 4-O-?-D-glucopyanoside were fistly separated from S.alopecuroides.By separating the chemical constituents of S.alopecuroides,it further enriches the chemical composition of S.alopecuroides and provides a material basis for its full development and utilization.Part 3 Study on Preparation Process of sophoridine and 13,14-dehydrosophoridine1.Establishment of a method for determination of the content of sophoridine and 13,14-dehydrosophoridine in S.alopecuroides.Firstly,an HPLC method is estabilshed for determinating content of sophoridine and 13,14-dehydrosophoridine and to optimized the materials from different districts.The results showed that the specific content,precision,stability and repeatability met the requirements,and the herb of Inner Mongolia was selected from the four origins.2.Extraction process.The best level of material/liquid ratio,extraction time,ethanol concentration and extraction times were screened by single factor test,and the exteaction of sophoridine and 13,14-dehydrosophoridine was optimized with Box-Behnken experiment Through comprehensive analysis,the optimized parameters were as follows:the material/liquid ratio was 1:11,the ethnol concentration of ethanol was 60%,and decocting frequency was three times,eachtime for 1.5 h.The respectively extraction rates of sophoridine and 13,14-dehydros-ophoridine were 86.14%and 83.09%.3.Enrichment and purification process.By screening six different types of macroporous resins,H103 macroporous resin was selected as a enriched filler of sophoridine and 13,14-dehydrosophoridine,meanwhile,the sample pH,sample loading,volume of water,eluent,pH and volume of eluent were investigated.The best enrichment process the macroporous resin as follows:sample of 0.5 g/mL with the pH=10 was absoipted to resin coloumn.After the sample was adsorbed,using water to wish 2 BV,then using 20%ethanol to wish 3 BV,finally,using 80%ethanol with pH 11.0 to wish 10 BV,collected 80%ethanol eluate,and the solvent was recovered under vacuum to obtain extract.The silica gel purification process as follows:using dichloromethane-methanol-ammonia water(5:0.04:0.02)as eluent,wet silica gel is packed into chromatographic column(diameter/height 1:8).Loading sample of mixed with silica gel into chromatographic column sampling at 1:20,then using eluent to elute,the mixture of sophoridine and 13,14-dehydrosophoridine will get.Crude products are reflux extracted by petroleum ether.The crude mixture is hydrogenated to obtain sophoridine.Another crude mixture is separated by silica gel and recrystallized to obtain 13,14-dehydrosophoridine.4.Purity test.The purity of the prepared sophoridine and 13,14-dehydrosophoridine was tested by TLC and HPLC at different unfolding systems and detection wavelengths.The results showed that respectively purity of sophoridine and 13,14-dehydrosophoridine is above 98.5%.