| Atractylodis Rhizoma origin from dried rhizome of Atractylodes lancea(Thunb.)DC.Studies have shown that there are some differences in the yield of the aqueous extract of Atractylodes lancea(A.lancea)from different habitats,and the sacchrides is the main component of the aqueous extract.In order to explore the differences in the saccharides of A.lancea.In this study,the total saccharide,oligosaccharides and crude polysaccharides were explored,and a quality evaluation method for oligosaccharides and crude polysaccharides of A.lancea and a prediction model for the total saccharide content of A.lancea were established.This study will provide a reference for the quality evaluation of saccharide from A.lancea.In addition,crude polysaccharide is one of the effective components of A.lancea to invigorate the spleen.In this study,the mechanism of crude polysaccharide of A.lancea for the treatment of spleen deficiency and diarrhoea from the perspective of intestinal flora will be further elaborated.The specific study methods and findings are as follows:1.The total saccharide content of A.lancea of different habitats was determined by phenol-sulphuric acid method and a prediction model based on hyperspectral technique was established for the total saccharide content.The results showed that the total saccharide content of A.lancea from different habitats ranged from 28.06%-34.72%.The highest total saccharide content of A.lancea was found in Hubei,Jiangsu Maoshan and Henan,reaching over34.08%,with significant differences compared to Anhui.The results of the random forest algorithm-based regression model of total saccharide content and spectral data of A.lancea showed a strong correlation between visible infrared spectral data and total saccharide content,with R~2 up to 0.857 1 and0.751 1 for the training and prediction sets of the established model,respectively.2.A method for the quantification and characterization of oligosaccharides in A.lancea was established based on ultra-performance liquid chromatography-quadrupole-time-of-flight masss pectrometry(UPLC-Q-TOF-MS)and ultra-performance liquid chromatography-evaporative-light-scattering detector(UPLC-ELSD),and the oligosaccharides in A.lancea from different habitats were quantified.The study showed that the geo-authentic characteristics of Jiangsu Maoshan were significant,with the peak area ratio of FF to GF oligosaccharides being greater than 1,while the other habitats did not have this characteristic.The total GF oligosaccharides content was highest in Henan,Jiangsu Maoshan and Hubei,reaching 18.51%,16.47%and 14.11%respectively.There was no significant difference in the content of the nine oligosaccharides between the two origins of Henan and Jiangsu Maoshan.The results based on principal component(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)showed that Henan and Jiangsu Maoshan clustered into two categories with other habitats respectively,and sucrose,GF2,GF3,GF4 and GF5 were the differential components between the two categories.3.A comparative study on the extraction of crude polysaccharides from A.lancea by different extraction methods showed that the structure of crude polysaccharides extracted by orbital shakers is consistent with that of water extraction and the method is relatively stable,which is a green and efficient extraction method.The main influencing factors of the orbital shakers extraction process were the material-to-liquid ratio and extraction time.The analysis of crude polysaccharides from different habitats of A.lancea based on multi-level structural characterization of infrared spectroscopy,molecular weight distribution,and monosaccharide composition showed that the structural differences between the geo-authentic and other production areas were not significant.The infrared spectra of the crude polysaccharides of A.lancea from different production areas were highly similar.High performance molecular exclusion chromatograms(HPSEC)showed that the profiles of Henan and Jiangsu Maoshan were not consistent with those of the other producing areas,with the Mw1 fraction being higher in Henan and Jiangsu Maoshan,at 1.85%and 1.35%respectively.The molecular weight distribution results of different habitats showed that the relative contents of Mw2 and Mw3 in the Henan differed significantly from other habitats,and the difference in fraction content may be one of the differences in crude polysaccharides in different habitats of A.lancea.The crude polysaccharides of A.lancea consisted of Fru,Man,Glu A,Rib,Allose,Rha,Gal A,Api,Glu,Gal,Xyl,Ara and Fuc,and for the first time,Rib and Allose were detected in the crude polysaccharides of A.lancea.The monosaccharide composition of the crude polysaccharides from different habitats of A.lancea was not significant except for the Glu content,while the content of the other 12 monosaccharides did not differ significantly.Based on the multivariate statistical analysis of the monosaccharide composition of crude polysaccharides from different habitats of A.lancea,the results showed that the clustering analysis based on three principal components could classify Anhui and Chongqing with Hubei and Henan,and the main different monosaccharide species were Fru,Glu and Allose.4.This study investigated the effects of geo-authentic A.lancea crude polysaccharide in the treatment of spleen deficiency diarrhea and explored its mechanism of action based on intestinal flora.The study showed that the crude polysaccharide of A.lancea could reduce the basic physiological indexes such as fecal water content,significantly increase the weight of immune organs and enhance the immunity of mice;meanwhile,the crude polysaccharide of A.lancea could restore the levels of GAS,MTL and D-Xyl in mice,and enhance the gastrointestinal motility and absorption function.The above study indicates that A.lancea crude polysaccharide is an effective biomolecule for the treatment of spleen deficiency and diarrhea.The results of the intestinal flora analysis showed that low doses of A.lancea crude polysaccharide could reverse the intestinal flora disorder caused by spleen deficiency and diarrhoea.Microbial community analysis at the phylum and genus level showed that Helicobacter abundance was suppressed and the abundance of beneficial genera,such as Akkermansia and Alistipes,was increased. |
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