高速逆流色谱分离纯化枸杞多糖在神经炎症中的作用OA北大核心CSTPCD

Objective:This study constructed a novel method for the separation and purification of polysaccharides from Lycium barbarum polysaccharide(LBPs)by high-speed countercurrent chromatography with aqueoust two-phase system,and investigated its effect on neuroinflammation.Methods:Through the partition coefficient(K)value and stationary phase retention rate Sf,the conditions for separating and purifying LBPs using aqueous two-phase system coupled with high-speed countercurrent chromatography(HSCCC)were screened and optimized.Preliminary structural characterisation of LBPs was performed using high performance size exclusion column-multiangle laser light scatter-refractive index detector(HPSEC-MALLS-RID),1-phenyl-3-methyl-5-pyrazolone precolumn derivatization coupled to high performance liquid chromatography(HPLC),Fourier transform infrared spectroscopy(FTIR),thermogravimetric analyzer(TG)and scanning electron microscopy(SEM).Moreover,the anti-neuroinflammation effect was investigated on LPS induced BV2 cell model.Results:The optimal biphasic system consisted of ethanol,ammonium sulphate,water and isopropanol had a ratio of 1.4∶1.0∶2.58∶0.3.Under the separation conditions(900 r/min and 0.8 mL/min mobile phase flow rate),three polysaccharide fractions,named LBPs-l,LBPs-2 and LBPs-3,was isolated.Polysaccharide contents were 78.35％±1.52％,69.03％±1.71％and 62.11％±2.31％.Molecular weights were 7.40×104 Da,2.73×104 Da and 1.47×104 Da.The monosaccharide composition was as follows:Man∶Rha∶GalA∶GlcA∶Glu∶Gal∶Ara,with the molar ratios of 1.9∶4.9∶1.3∶8.1∶7.6∶32.6∶41.8,2.1∶4.3∶1.5∶8.3∶5.1∶28.6∶37.9 and 5.9∶4.6∶1.1∶0.7∶13.8∶66.8∶5.3,respectively.There was also a significant difference in terms of thermal stability and micro-morphology.The LBPs exhibited varying degrees of protective activity on LPS-induced BV2 cells.Conclusions:This study would provide technical support for the efficient separation and purification of plant polysaccharides,and scientific basis for the application of LBPs in the inhibition of neuroinflammation related diseases.