Rapid Separation And Analysis Of Ginsenoside Based On Multi-Walled Carbon Nanotubes

Objective: Ginseng is known as the "king of herbs" with a long history of medicinal use.Ginseng contains a variety of pharmacological active ingredients such as ginsenosides polysaccharides,amino acids and so on.Ginsenosides are the main pharmacological active components,which are widely used.In this paper,the ginsenosides in the ginseng crude extract were extracted with porous carbon materials,and the multi-response detection mode(MRM)in HPLC-MS technology was used to detect them,and the extraction efficiency of ginsenosides in the ginseng crude extract with porous carbon materials was analyzed.Ginseng quality varies,and ginseng from different places of origin has different quality.At present,there is no good evaluation method.In this paper,the HPLC-MS method is established,the multi-response detection mode(MRM)of HPLC-MS method combined with the multivariate statistical method is used to screen saponins,and the ginseng places of origin are distinguished by the content of ginsenosides.Methods: 25 mg MWCNTs were added to 5 ml 200 mg/ ml ginseng crude extract aqueous solution,which was slightly shaken for 2 s,centrifuged at 8000 RPM for 2 min,and precipitated with MWCNTs for the desorption of ginsenosides.The supernatant was filtered with 0.22μm water filtration membrane in a 10 ml volumetric flask,and the contents of 8 gensenosides were determined by HPLC-QqQ/MS 5ml of saturated n-butanol was added to the centrifuged MWCNTs,which were slightly shaken for 5 s and centrifuged at8000 RPM for 1 min.The supernatant was repeated for 3 times.The content of 8ginsenosides was detected by 0.22μm water filtration membrane as the previous step.The content of sugar in ginseng crude extract before and after extraction was determined by phenol-sulfuric acid method so as to determine the extraction efficiency of ginsenosides by multi-walled carbon nanotubes.Ginseng drying to constant weight after washing,crushing,will smash the powder had 40 mesh points stipulated by the pharmacopoeia sample sieve,in 4 ℃ dry environment preservation.Take 1 g ginseng powder and 50 ml ether,add to the soxhlet extractor reflux 2 h under 40 ℃,the purpose in order to remove the fat solubleginseng powder;Refuse to ether liquid,and then precision add 50 ml methanol circumfluence extraction 2 h,will extract in rotary evaporation under 40 ℃ to dry after dissolved in 10 ml water,saturated with 30 ml water extraction 3 times in a row,n-butyl alcohol solution aims to remove strong polarity ingredients such as sugar;Will consolidate merger extract,extract in 40 ℃ rotary evaporation to dry,dissolved in methanol and constant volume to 10 ml;After 0.22μm quartz fiber filtration membrane,hplc-ms analysis was performed.The obtained data were combined with the multivariate statistical analysis method to screen ginsenosides and determine the origin of ginseng by analyzing the content of ginsenosides.Results: MWCNTs can adsorb most ginsenosides,but have no obvious adsorption effect on sugar,indicating that MWCNTs can separate the sugar and ginsenosides in the extract of ginseng.MWCNTs has a good adsorption capacity for 8 main ginsenosides and can achieve the equilibrium of adsorption and desorption in a short time.The adsorption capacity of the original ginsenosides Ro of ginsenotriol and oleanolic acid is slightly higher than that of the original ginsenosides.Water-saturated n-butanol solution could rapidly elution the ginsenosides adsorbed by MWCNTs,and the recovery of ginsenosides increased with the volume of elution solvent.When the volume of elution solvent was above 20 mL,the recovery of 8 ginsenosides was higher than 85%.The species of ginsenosides in ginseng from 5 different producing areas were different,and the species and content of ginsenosides in similar producing areas were similar.Forty-one ginsenosides were identified by tandem mass spectrometry and polarity difference of ginsenosides.Using multivariate statistical analysis technique and hplc-ms full scan data of ginsenoside extract,a model was established to distinguish ginseng from 5 producing areas of suihua city,xunke county,hulin city,changbai county and wangqing countyConclusion: The use of porous carbon materials for rapid extraction and separation of ginsenosides in the crude extract of ginseng,compared with traditional methods,has the advantages of solvent saving,time saving and simple operation,etc.Compared with the separation method of macroporous resin,MWCNTs has the same separation effect andfaster separation speed.The quality of ginseng is different,among which the producing area has a great influence on its quality,and the growing years have a certain influence on its quality,but the influence is not big.