The Preparation And Antibacterial Activity Of Less Polar Ginsenosides From Panax

American ginseng?Panax quinquefolius?,Asian ginseng?Panax ginseng?and notoginseng?Panax notoginseng?are ginseng botanicals which have been used for thousands of years as important resources of health food all over the world.Saponins in ginseng,also called ginsenosides,are considered to be the main biological constituents.According to the polarity of the ginseng saponin,ginsenosides can be divided into polar ginsenosides and less polar one.The ginsenosides in nature plant is porar one.Less polar ginsenosides have strong biological activities than the polar.However,therer was no transformation is more conducive for application.According to the data of World Health Organization,the incidence of diseases?dermatophytes,halitosis?has a high prevalence caused by microorganism.Exploring the active ingredient from natural products for treating related diseases is hot in recent years.However,the research on antimicrobial effect of ginsenosides was rare.This study intends to get the less polar ginsenosides from Panax,investigate the antimicrobial?dermatophytes,halitosis?activity and mechanism of ginseng saponins,develope bacteriostatic effect of toothpaste and wash liquid products,respectively.The main research and work in this paper involves as follows:?1?The different varieties,different parts of the low polarity saponins of conversion and loss ratio of ginseng total saponins were investigated systemly.It was found that American ginseng stem leaf and notoginseng total saponin is suitable for the material as the conversion of low polarity saponins.The contents of ginsenosides Rb2,Rg2?S?and Rd abundantly present in American ginseng stems-leaves saponins?AGS,0.81±0.003 mg/mg?were significantly transformed into less polar one at the rate of84%.The less polar ginsenosides in AGS and HTS were 0.08±0.005 mg/mg,0.76±0.04 mg/mg.NS contained five kinds of polar ginsenosides?0.66±0.06 mg/mg?.However,after heat treatment,these ginsenosides were transformed into the less polar ginsenosides of Rk3,Rh4,and Rh5 in TNG at the rate of 90%and the less ratio of ginsenosides was 0.9%.?2?In this study,19 compounds were isolated from the transformed American steam-leaf saponins by solvent extraction,repeating column chromatography and recrystallization.By the spectral analysis and chemical evidence they were identified as follows:ginsenosides 20?E?-Rh4,20R-Rg3,Rk3,20?z?-Rh4,Rh2,20?z?-Rg6,F4,20?S?-Rh2,20?S?-Rg3,20S-Rg5,Rs3,Rk2,20S-Rg2,20R-Rg2,Rg1,Rc,Rd,Rb2,20?S?-Rh,protopanoxadiol.?3?The antibacterial activity of ginsenosides was investigated by MIC and MBC.AGS was not detected even when the concentration was at 2.5 mg/mL.C.perfringens and P.gingivalis were mostly susceptible to HTS-4,with the MIC of 9.8?g/mL and MBC of 312.5?g/mL.In our test,less polar ginsenoside fractions HTS-4 demonstrated their highest activity against standard strains of anaerobic bacteria.Among the three fungi,T.rubrum was most susceptible to HTS,with the lowest MIC.The MIC of HTS against T.rubrum was just doubled than that of miconazole nitrate,which implied that HTS could be also used as a potent antifungal agent.The experiments show less polarity saponins are superior to the polarity saponins.?4?Further more,the effect of ginsenosides on the integrity of microbial membrane system were explored by the content of protein and nucleic acid from outside of membrane,fluorescent confocal microscopy,transmission electron microscopy,CBD plate method,the bacteria produce acid was investigated using GC/MS.The absorbance value for nucleic acids(OD_260nm)of P.gingivalis increased significantly?P<0.05?from 0.047 for the control to 0.100 and 0.184 with the presence of HTS-4 at the levels of MIC and MBC at 22 h,respectively.After 18 h,OD_260nm value of P.gingivalis almost stop to growth.Under similar conditions,there was a progressive release of nucleic acids of C.perfringens and F.nucleatum for up to almost 18 h followed by a steady state.Moreover,values for proteins also showed significant increase?P<0.05?for both bacteria.The values of proteins at MBC levels increased faster than that of MIC levels.After addition of HTS-4 at MBC level,a rapidly decrease were occurred in all bacteria with the MFI values decreased 77.21,64.53,and 78.7 percent compared with control,respectively.Furthermore,the MFI of three bacteria treated with HTS-4 at MBC level decreased by57.66,25.85 and 40.54 percent respectively.Confocal fluorescence microscopy and CBD plate method validation of biofilm in low concentration has been suppressed,high concentrations of ginseng directly by low polarity saponins can kill bacteria biofilm.The result from GC/MS was founded that under the MBC level of nuclear fusobacterium capacity of acid inhibition by almost 90%.?5?Finally we produced a series of toothpaste and contain gargle fluid respectively,as a product display.