| Pseudoginsenoside F11(PF11)is a unique component in American ginseng,and its distribution in roots,stems,leaves,flowers and fruits,it is the highest in stem and leaf.Its low toxicity,strong pharmacological activity,the current composition of a wide range of research,mainly in the separation preparation,pharmacology research,dosage form development stage.In the molecular,receptor,cell and the whole level of pharmacological pharmacological studies have great potential value,attracting the interest of the majority of drug researchers.However,due to the low amount of PF11 naturally occurring,and in the total saponin by other components,physical and chemical properties of complex,is not conducive to the separation of the target compound.In this paper,the preparation of PF11 was carried out by using extraction method,precipitation method and chromatography method.The specific process was as follows:Design extraction process for separating PF11.Take a certain amount of American ginseng leaf saponins,dissolved in water.The content of PF11 in the separation process was determined by HPLC-ELSD chromatography.It was found that PF11 was mainly enriched in the n-butanol fraction.The effects of extraction time,extraction temperature,saponin concentration and extraction time on the separation of PF11 were investigated by orthogonal test.The optimum separation conditions were obtained.The results showed that 100 g of saponin was dissolved in 500 ml of water at 40 ° C and extracted twice with 250 ml of n-butanol to take the n-butanol fraction.This process is simple and easy to implement.Design of precipitation method for s solution and the mixture is added with stirring to change the polarity of the n-butanol partial solution and precipitate a part of the saponin.The content of PF11 in n-butanol-acetone precipitate was determined by HPLC-ELSD chromatography.PF11 was mainly distributed in the filtrate after precipitation.The effects of precipitation rate,precipitation time,precipitation time and volume ratio of n-butanol to acetone on separation of PF11 were investigated by orthogonal experiment.The preliminary separation of the target compound PF11 was carried out.As a result,800 ml of n-butanol was partially added to the acetone solution at 30 ° C,precipitated once and allowed to stand for 5 min.The optimum conditions for the separation of PF11 were investigated by acetone-n-butanol-saturated sodium chloride and orthogonal test.The effects of precipitation temperature,precipitation time,precipitation time,liquid volume ratio of mixture liquid and acetone were investigated.The filtrate was concentrated,evaporated to dryness,and dissolved in a mixed solution of 600 ml of n-butanol-acetone-saturated sodium chloride(1: 4: 1 by volume).The mixed solution was added to 3L of acetone solution,Set 5min,filtration.Acetone in the salt solution to change the polarity of the mixture to achieve part of the separation of saponins.In the filtrate part of the dry,weighed,PF11 crude 30 g,content of 5.1%.The contents of six saponins(Rb1,Re,Rd,Rg1,Rb2,Rc)were determined during the separation process.The results showed that the contents of six saponins in n-butanol extraction solution were very few Butanol has a strong ability to dissolve saponins.After extraction and precipitation separation process,the content of PF11 was enriched from 2.24% to 5.1%,and the enrichment was achieved.It was also found that the purity of ginsenoside was enriched from 14.0% to 28.4%,while ginsenoside Rg1 was well enriched.Design chromatography purification process route.First,the adsorption and desorption of saponin by macroporous resin XAD-2 were investigated.The first step was to purify PF11 with XAD-2 macroporous resin.A first step of purification of 122.9 g crude PF11(5.1%)was carried out using the XAD-2 macroporous resin method.The dynamic loading of PF11 was detected by HPLC-ELSD and TLC.The optimum conditions were determined by the amount of macroporous resin,the amount of sample concentration,the alcohol content of the eluted saponin and the elution rate.Purified by macroporous resin once to achieve PF11 purity of 11.3%,the first step to separate PF11 crude 45.3g.Followed by a second step of purification of 40 g of PFO crude(purity: 11.3%)using C18 reverse phase column chromatography and recrystallization.The mobile phase was determined by TLC method as methanol: water(9: 5).The optimum conditions were determined by measuring the loading concentration,flow rate and eluent amount of column chromatography(C18 filler).After column chromatography(C18 filler),the sample was recrystallized from ethyl acetate to give 3.67 g of sample having a purity of 90.3% of PF11 in a yield of 40.5%. |
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