Study On Consititues Of Alkaloids In The Seeds Of Crinum Asiaticum L.

Crinum asiaticum L.is Crinum genus of Amaryllidaceae family,which is a member of perennial herb compositae,whose seeds are round and yellow when matured.It exists mainly in the south of China.All plants of Crinum asiaticum L.can dispel wind and dampness,eliminate swelling.They have rich chemical composition such as volatile oils,flavonoids and their glycosides,alkaloids and glycosides,fatty,terpenoids,steroids,amimo acides and so on.In clinic,their chemical composition is used as a traditional medicine to relieve pain,activate blood circulation,treat bone-stasis swelling,fractures,poinsonous snake bites and so on.Crinum asiaticum L.bulbs and leaves contain a lot of antitumor active alkaloids ingredient,the chemical constituents of seeds with biological activity has not been reported.To confirm its chemical composition and related pharmacological activity and provide a theoretical basis for the further development and utilization of Crinum asiaticum L.This paper studied the chemical composition of alkaloids in the fruits of Crinum asiaticum.Single factor experiment and response surface methodology were used to optimize the best extracted way and the extraction process.The extracts were used acidic water-alkaline-simple impurity extraction method to remove impurities according to the characteristics of alkaloids.Dichloromethane was used as the extraction solvent to obtain total alkaloid extracts.Then the total alkaloid extracts were separated and purified by multiplty chromatography.The sample compounds were detected by TLC,in which the compounds in same Rf values were mergered.The mergered compounds that their was analyzed by HPLC,in which the chromatographic parameters were set as follows:C18 column,mobile phase of methanol-water,flow 1.0mL/min.Record the mobile phase proportion when the compounds peaks were baseline separated.Under this chromatographic condition,a fraction can be separated into several pure compounds by a C18 column.The structures of the isolated monomeric compounds were finally identified by NMR and MS technnlogy.The heating refluxing extraction method was used,and optimization results by response surface method are as follows:extraction time 2.5 h,extraction temperature 84? material liquid ratio 1:9,ethanol concentration of 95%,highest alkaloid extraction rate 0.4494%.The total alkaloids were extracted according to this optimized method.The crude alkaloidwas separated using silica gel column chromatography,Sephadex LH-20 chromatography,HPLC analysis and preparation to get three pure compounds.Their structures were elucidated as follows:(1)N-trans-feruloyl tyramine,(2)liquiritigenin,(3)hippacine.