Study On The Purification And Extraction Of The Immunomodulatory Effective Part Of Panax Quinquefolium Total Saponins, Total Polysaccharides Of Yi Yuan Kang Fang

Objective: In recent years, there are many research about American ginseng to prove that it has a wide range of biological activities, the main active ingredient of ginseng total saponin and polysaccharide compounds can act on a variety of immune cells, promote the secretion of certain cytokines to further exert immunomodulatory effects. This topic will extract and purificate these which bioactive,such as the total saponin and total polysaccharides and other components about American ginseng,as Kang Yi Yuan Fang further screening effective components research and immunomodulatory effects of basis.Methods: In medicine quality control, plus 80% ethanol 50 ml, heat reflux extraction three times, each time 1h American ginseng extract total saponins, vanillin using acetic acid as chromogenic reagent, using UV-visible spectrophotometry total saponin content. Add water 50 ml, water bath was extracted three times, each time 1h, filtration, combined filtrate was concentrated to 50 ml, the precise amount of 10 ml, adding ethanol 30 ml, shake, 4 ℃ standing 24 h, removed by centrifugation(4000r?min- 1,10min), the supernatant was decanted, the precipitate dissolved in water, adding an equal volume of Sevag reagent(n-butanol: chloroform = 1: 4) in addition to protein twice to give a total for the test polysaccharide, anthrone sulfuric acid method significantly Determination of total polysaccharide color.Through the 12 batches of Panax quinquefolium herbs fingerprint map construction, compared to the reference of the retention time of the seven peaks chemically identified, using fingerprints and similarity parameters common peaks were analyzed, and map cluster analysis. The effective part of the extraction process in order Panax quinquefolium total saponins, ginsenoside Rg1,Re, Rb1 as an indicator, the single factor experiment, even experimental study of Panax quinquefolium total saponin alcohol extraction process; to Panax quinquefolium as an indicator of total polysaccharides, single-factor test, orthogonal experimental study of Panax quinquefolium total polysaccharides extraction process. In the process of research and purified by macroporous resin for different types of adsorption and desorption properties of the study, The optimum resin adsorption, and were single factor test, test, and even single-factor test, orthogonal test Panax quinquefolium total saponins and purification of process parameters total polysaccharides.Results:(1) quality control of medicines, ginsenosides Rg1, Re, Rb1 content of more than 2.0% of total Pharmacopoeia, controllable quality. And the establishment of Panax quinquefolium HPLC fingerprint, marked a total of 30 peaks, the similarity in the 0.925 to 0.995,12 approved medicinal clustered into four categories.(2) Panax quinquefolium total saponin alcohol extraction process: the use of 12 times the amount of 85% ethanol extract twice 160 min, combined extract was filtered and concentrated. Panax quinquefolium total polysaccharides extraction process: liquid ratio of 10: 1, extracted three times, each time 1.5h, combined extract was filtered and concentrated.(3) were selected D101, HPD600 resin purification as the best Panax quinquefolium total saponins and total polysaccharides, preferably for the purification process of total saponins: the sample concentration was 14.84mg?ml-1, the sample flow rate was 1.0ml?min-1, diameter higher than 1:10, the volume of washing 3BV, eluent of 80% ethanol, the eluent in an amount of 200 ml,the purity of American ginseng total saponins is 57.45%.The total polysaccharide purification process: the sample concentration was 3.64 mg?ml-1, PH 8.0, the sample flow rate of 2.0 ml?min-1, diameter and height ratio of 1: 8, the maximum loading capacity of 3.5BV, wash removal agent is 20% ethanol, the eluent PH 8.0, eluent dosage 4BV,the purity of American ginseng total Polysaccharide is 47.94%.Conclusion:By Panax quinquefolium total saponins, total polysaccharides Extraction and purification process, the extraction and purification process is feasible to obtain total saponins, total polysaccharides stable and controllable quality.