Mass Spectrometry Analysis And Neuroprotective Effects Of Ginseng Oligosaccharides

Objective: We established a method for the analysis of seven types of ginseng oligosaccharides using ion mobility mass spectrometry(IM-MS)and liquid chromatography-mass spectrometry(LC-MS),achieving qualitative and quantitative analysis of oligosaccharides in ginseng,and determining differences between them.We analyzed the role of American ginseng(AG)and black ginseng(BG)oligosaccharides in improving cognitive impairment through mice models of scopolamine-induced cognitive impairment for learning.Methods: In this study,oligosaccharides in fresh ginseng(FG),red ginseng(RG),AG,dried ginseng(DG),black ginseng 1(BG1),black ginseng 2(BG2),and black ginseng 3(BG3)were compared and analyzed by IM-MS.The distinct components of several ginsengs were identified using statistical analysis and three-dimensional IM-MS.The oligsaccharides in the seven varieties of ginseng were quantitatively analyzed using liquid chromatography-mass spectrometry(LC-MS)in multiple reaction monitoring(MRM).The data were processed using the principal component analysis method,and the seven different types of ginseng oligosaccharides were compared and examined for content differences.The learning and memory capacities of the mice with cognitive impairment were assessed using the Morris water maze test and the novel object identification test.By using an enzyme-linked immunosorbent assay(ELISA),the levels of acetylcholine(Ach),acetylcholinesterase(Ach E),glutathione(GSH),malondialdehyde(MDA),and superoxide dismutase(SOD)were measured in order to assess the effects of AG and BG oligosaccharides on the cholinergic nervous system and oxidative stress in the cerebral cortex of mice with cognitive impairment.The scopolamine-induced cognitive impairment mice were used as the model,and the experiment was divided into 7 groups: The control group(CON)and model group(SCOP)were given normal saline daily by gavage.The daily doses of donepezil positive drug group(DON+SCOP),AG oligosaccharide extract low-dose group(AGOSL+SCOP),AG oligosaccharide extract high-dose group(AGOSH+SCOP),BG oligosaccharide extract low-dose group(BGOSL+SCOP),and BG oligosaccharide extract high-dose group(BGOSH+SCOP)were 1,40,80,40 and 80 mg/kg,respectively.On the sisth day,the mice were made membranes,and after 30 minutes of intragastric administration,the CON group was intraperitoneally injected with normal saline,and the other groups were intraperitoneally injected with 1.5 mg/kg scopolamine.After 30 minutes,the behavioral tests were performed.At the end of the behavioral test,the cerebral cortex of the mice was collected for ELISA.Results: The seven ginseng samples shared fructose,glucose,maltose,maltotriose,and maltotetraose,according to the IM-MS study.Only three types of BG included unknown derivated-oligosaccharide ions of m/z 391.18,m/z 420.21,and m/z 443.20,and only fresh ginseng contained isomaltotetraose and only red ginseng contained an unknown tetrasaccharide.All other ginseng varieties—aside from fresh ginseng—contain unknown derivated-oligosaccharide ions of m/z 383.16 and m/z 385.14.The molecular formula for the five unidentified oligosaccharides discovered in ginseng was determined,and it was assumed that the structure contained alkene functional groups,esters,or amino groups.Multivariate statistical analysis results showed that seven ginseng varieties were divided into two groups(for the purpose of this work,named group I and group II).Group I comprised RG,DG,AG,and FG.Group II contained BG1,BG2,and BG3.The two types of ginseng included differing amounts of m/z 385.14,m/z 391.19,m/z 421.21,m/z 420.21,maltose,and maltotetracose.The results of behavioral experiments showed that both AG oligosaccharides and BG oligosaccharides could improve the learning and memory ability of scopolamine induced cognitive impairment mice,and the effect of AG oligosaccharides was better than that of BG oligosaccharides,and there was no dose-response relationship.The results of ELISA experiments showed that AG and BG oligosaccharides improved spatial learning and memory ability by increasing the level of Ach in the cerebral cortex of cognitively impaired mice,reducing oxidative stress in the brain by regulating the levels of SOD,GSH and MDA,and improving learning and memory abilities.In regulating SOD levels,the effect of BG oligosaccharides is better than that of AG oligosaccharides.Conclusion: Qualitative and quantitative approaches were developed for the analysis of oligosaccharides in DG,RG,AG,FG,and three types of BG by applying IM-MS and LC-MS procedures.The chemical formula and potential structural formula of five unknown compounds were hypothesized.The distinctive components were discovered using multivariate statistical analysis.It serves as a resource for the ginseng pharmacodynamics study,quality assessment,and processing procedure.In improving spatial memory and regulating SOD levels,the effect of BG oligosaccharides was better than that of AG,and there was no dose-effect relationship.There is a dose-dependent in the regulation of GSH and there is no dose-dependence in the regulation of other indicators.AG and BG oligosaccharides can improve cognitive impairment in mice by regulating cholinergic neurotransmitters and oxidative stress markers.In improving spatial memory and regulating SOD levels,the effect of BG oligosaccharides was better than that of AG,and there was no dose-dependence.There was a dose-dependent regulation of GSH,and there was no difference between the AG and BG oligosaccharides.This study provides a reference for the pharmacodynamic study of cognitive impairment diseases in BG and the development of food and health products.