Studies On Chemical Component And Extraction Process From The Seeds Of Impatiens Balsamina L.

The seeds of Impatiens balsamina L is a traditional Chinese medicine. It is effect on the soft-firm, digestion block, feature bone, hard throat and so on. It have stimulant transdermal, anti-fertility, anti-tumor, anticancer, antibacterial effect. Also, it can be used in clinical practice contraception, termination of pregnancy and inhibit in esophagus and cardia tumor and bone hyperplasia.The seeds of Impatiens balsamina L contains so many compounds such as flavonoids, volatile oil constituents, saponins, of which the highest content is the hosenkosides. In this experiment, four kinds of main hosenkosides,hosenkosides K, A, C, B were used as the indexes of total hosenkosides extraction from the seeds of Impatiens balsamina L. The quality standards of total hosenkosides also have been studied, and the results can provided the theoretical basis for pharmacology in the future.The first part of this paper is talking about the separation of six hosenkosides isolated from the seeds of Impatiens balsamina L.. The six hosenkosides were identifed as hosenkoside K, hosenkoside A, hosenkoside C, hosenkoside B, hosenkoside D and hosenkoside E, of which four of them hosenkoside K(H1),A(H2),C(H3) and B(H4) were used as the indexs for further experimental study.In the second step of the experiment, in order to determine the best extraction condition that the total saponins were extracted from the seeds of Impatiens balsamina L, the solvent concentration, the ratio of material to liquid and reflux time were tested using orthogonal test. The content of four compounds H1, H2, H3, H4 were determined by HPLC in each condition as indexes.The results showed that optimal condition were that the seeds of Impatiens balsamina L, were extracted by 70% ethanol (V/W is 6 times), refluxed 4 times-60min,45min,30min,30min respectively,In the further experiment, this paper investigated the static adsorption and static desorption of 9 different types of macroporous resin(HPD-100,D101,D4020, ADS-17,S-8,DA-201,NKA-9,DM130,AB-8), the results showed that the AB-8 macroporous resin was the best choice and played a good separation efficiency.The pH of the solution,flow rate,elution agent factors were investigated and eventually decided the macroporous resin purification parameters.The color of the solution washed from the macroporous resin is so heavy that the decoloration resin is needed.In this part 5 kinds of decoloration resin (D315,D310, D941,D280,330) were selected, and the final selection is D310. At the same time this paper have been investigated the adsorption time,adsorption temperature effected on the rate of the decolorization,the result is that the ethanol elution flew the decoloration resin chromatographic column at room temperature and decolorded for 3 hours.The total saponins obtained from the macroporous resin and decolorization resins still contains more impurities,it is required to be further purified by silica gel column.chromatography. The parameters of the ratio of sample to silica gel(w/w) and mobile phase were tested.Based on the above experimental results, the final purification steps were as follows:The crushed seeds of Impatiens balsamina L, were extracted with 6 times(V/W) of 70%ethanol and refluxed four times, the extraction time were 60min,45min,30min, 30min respectively. The solution were combined after filtered and evaporated to the 6 times of the material wight in vacuum. The solution (pH=6.0) was through the AB-8 macroporous resin, then eluted by 80% ethanol for 6 times of the resin volume. All eluting solution was through D310 decolorization resin. The crude saponins were obtained after the solvent have been evaporated in vacuum. The crude saponins were subjected to silica gel column chromatography and eluted by ethyl acetate-ethanol-water (4:1:0.8), the ratio of silica gel to sample is 25:1(w/w).The collected solution that eluted from the column were determined by TLC, all the solution containing total saponins were combined and evaporated in vacuum and were dried at 105℃for 3 hours, then the total saponins could be obtained. The contents of compound H1, H2, H3, H4 in the total saponins were determined by HPLC-ELSD and the sum of the content were higher than 50%with transfer rate of H1,H2,H3,H4 passed 80%.Besides the quality standard of total saponins from the seeds of Impatiens balsamina L was established. The water, residue on ignition, heavy metals testing, arsenic salt determination have been detected. The results showed that the samples dry weight loss was less than 2%, Residue on ignition less than 0.1%, heavy metal content and arsenic salt content were lower than the pharmacopoeia allowed. It also established the method to determine the total saponins of four kinds of major hosenkoside H1,H2,H3,H4 by HPLC-ELSD.The results showed that the mean of compound hosenkoside K is 38.66%, compound hosenkoside A is 5.21%, compound hosenkoside C is 1.23%, compound hosenkoside B is 8.25%and the total content is 53.35%. The determination method was accurate, precise, high sensitivity, wide linear range, and the mobile phase composition is simple.