Preparation Of Rare Ginsenosides And Mass Spectrometry Identification Research

Ginseng is a kind of traditional Chinese medicine with high medicinal and health functions.The main active components of ginsenosides are common ginsenosides and rare ginsenosides.Rare ginsenosides refer to the low or nonexistent ginsenosides in Panax plants.Most of them belong to the secondary ginsenosides produced by the removal of some glycosyl groups,or the secondary ginsenosides produced by the change of the side chain structure of the mother nucleus at the same time Physical activity.The preparation of rare ginsenosides by biotransformation with high specificity,stable reaction and environmental friendliness has become a hot spot in the field of health products.It not only changes the pollution problem of traditional chemical method,but also improves the safety of food and the controllability of preparation process.In this paper,rare ginsenosides were prepared by biotransformation of common ginsenosides in Panax quinquefolium by medicinal and edible bacteria,and then purified by macroporous adsorption resin to obtain rare Ginsenosides with high purity,which improved the application value of ginsenosides in medicine and health care products.This study first studied the effects of different factors on the extraction volume of ginsenosides,such as solvent concentration,material-to-liquid ratio,extraction temperature and ultrasound time.It was found that when the extraction conditions were1:40 with a material-to-liquid ratio,adding an ethanol solution with a concentration of80% and sonicating at 75°C for 60 minutes would result in a higher total saponin content.Two extraction verifications were carried out under this condition,and the total saponins extraction rates obtained were 5.528 mg/mL and 8.632 mg/mL,respectively,which exceeded the total saponins extraction rate in the condition optimization experiment,indicating that the optimal conditions were effective.In order to obtain a relatively pure and rare ginsenoside solution,a macroporous resin with better adsorption and desorption effects on the ginsenoside solution is selected.This experiment uses a combination of static adsorption and dynamic adsorption of primary and secondary selection.From D3520,S-8,D4020,D201 C,HP-20,X-5,HPD400,HPD450,AB-8,D101 C,D101,these 11 kinds of macroporous resins,gradually screened out the macroporous resin D101 with the best adsorption effect.Sample conditions and elution conditions were uniformly designed and optimized.The optimal solution was finally determined to be the saponin solution concentration of 8.27mg/mL,the sample volume(resin: volume)of 1:2,and the volume of the deionized water eluent.The volume of 64.3 mL,20% ethanol desorption solution was 128.6 mL,the volume of 60% ethanol desorption solution was 192.9 mL,and the volume of 100%ethanol desorption solution was 128.6 mL.Purification verification was performed on this condition,and the verification result was that the adsorption rate was 100% The deionization rates of deionized water eluent,20% ethanol desorption solution,60%ethanol desorption solution,and 100% ethanol desorption solution are 5.98%,-6.79%,77.49%,and 16.95%,respectively.Finally,the extraction liquid of solid fermentation of American ginseng and the eluate of purification by macroporous resin were identified by mass spectrometry.The variety and content of rare ginsenosides before and after solid fermentation and purification by macroporous resin were compared respectively,and our target rare ginsenosides could be finally obtained.In this study,the total saponins extract from solid fermentation of American ginseng and the saponins in the eluate purified by macroporous resin were identified by mass spectrometry.The types and contents of rare ginsenosides before and after solid fermentation and purification by macroporous resin were compared.It is found that after solid fermentation of American ginseng,new rare ginsenosides are produced,such as Rh1,20(R)-Ginsenoside Rh1,F1,Rh3,Rh4,Rh6,Rk3,Notoginsenoside R2,etc.;CK,Mc and Rd the content of the original rare ginsenosides has also increased.Among them,the content of CK has increased by 336.72%.This is because the conversion pathway of CK is the most extensive,and the types of hydrolases and prototype ginsenosides are abundant;because the rare ginsenosides are converted from common ginsenosides Therefore,the content of most common ginsenosides has been reduced,such as Rg1,Re,Rb2 and Rb3,Rc,etc.,which are reduced by 78.99%,91.8%,Rb2 and Rb3,which are isomers of each other.Separately,a total of 94.62% and 71.17% were reduced;and some saponins disappeared,such as Malonyl ginsenoside Rd,Malonyl ginsenoside Rc,Malonyl ginsenoside Rb1,Malonyl ginsenoside Rb2 and other propionyl ginsenosides and partially acetylated ginsenosides Such as Acetyl G-Rc,Acetyl G-Rg3,Acetyl G-Rg1.After purification by macroporous resin,the content of rare ginsenosides such as Rh1,F1,F2,Rk2,Rk3,Rg3,CK and other target products were enriched in the gradient ethanol eluent,and the enrichment degree of Rh1 was 10.02 times of the original solution,F1 The degree of enrichment was 34.39 times that of the original American ginseng fermentation broth,F2 was 1.31 times that of the original saponin solution.The degree of enrichment was only 1.8 times that of the original solution,but the CK content in the 100% ethanol eluate is slightly higher than half of the CK content in the original solution,indicating that the elution effect was better.Based on the above results,rare ginsenosides were prepared by solid-state fermentation of Panax quinquefolium,and the extraction conditions were optimized to increase the extraction rate,and then purified by the selected macroporous resin D101.