Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features.Therefore,quantitatively quality control of polysaccharides based on their chemical charac-teristics i...Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features.Therefore,quantitatively quality control of polysaccharides based on their chemical charac-teristics is important for their application in biomedical and functional food sciences.However,poly-saccharides are mixed macromolecular compounds that are difficult to isolate and lack standards,making them challenging to quantify directly.In this study,we proposed an improved saccharide mapping method based on the release of specific oligosaccharides for the assessment of Hericium eri-naceus polysaccharides from laboratory cultured and different regions of China.Briefly,a polysaccharide from H.erinaceus was digested by b-(1-3)-glucanase,and the released specific oligosaccharides were labeled with 8-aminopyrene-1,3,6-trisulfonic-acid(APTS)and separated by using micellar electrokinetic chromatography(MEKC)coupled with laser induced fluorescence(LIF),and quantitatively estimated.MEKC presented higher resolution compared to polysaccharide analysis using carbohydrate gel elec-trophoresis(PACE),and provided great peak capacity between oligosaccharides with polymerization degree of 2(DP2)and polymerization degree of 6(DP6)in a dextran ladder separation.The results of high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector(HPSEC-MALLS-RI)showed that 12 h was sufficient for complete digestion of polysaccharides from H.erinaceus.Laminaritriose(DP3)was used as an internal standard for quantifi-cation of all the oligosaccharides.The calibration curve for DP3 showed a good linear regression(R2>0.9988).The limit of detection(LOD)and limit of quantification(LOQ)values were 0.05 mg/mL and 0.2 mg/mL,respectively.The recovery for DP3 was 87.32(±0.03)%in the three independent injections.To sum up,this proposed method is helpful for improving the quality control of polysaccharides from H.erinaceus as well as other materials.展开更多
基金the National Natural Science Foundation of China(Grant No.:81673389)the Science and Technology Development Fund,Macao SAR,China(File Nos.:0075/2018/A2,034/2017/A1 and 0017/2019/AKP)+1 种基金the Guangdong Key Project for Modernization of Lingnan Herbs(Project No.:2020B1111110006)the Multi-Year Research Grant from the University of Macao(File Nos.:MYRG2018-00083-ICMS,MYRG2019-00128-ICMS,and CPG2022-00014-ICMS).
文摘Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features.Therefore,quantitatively quality control of polysaccharides based on their chemical charac-teristics is important for their application in biomedical and functional food sciences.However,poly-saccharides are mixed macromolecular compounds that are difficult to isolate and lack standards,making them challenging to quantify directly.In this study,we proposed an improved saccharide mapping method based on the release of specific oligosaccharides for the assessment of Hericium eri-naceus polysaccharides from laboratory cultured and different regions of China.Briefly,a polysaccharide from H.erinaceus was digested by b-(1-3)-glucanase,and the released specific oligosaccharides were labeled with 8-aminopyrene-1,3,6-trisulfonic-acid(APTS)and separated by using micellar electrokinetic chromatography(MEKC)coupled with laser induced fluorescence(LIF),and quantitatively estimated.MEKC presented higher resolution compared to polysaccharide analysis using carbohydrate gel elec-trophoresis(PACE),and provided great peak capacity between oligosaccharides with polymerization degree of 2(DP2)and polymerization degree of 6(DP6)in a dextran ladder separation.The results of high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector(HPSEC-MALLS-RI)showed that 12 h was sufficient for complete digestion of polysaccharides from H.erinaceus.Laminaritriose(DP3)was used as an internal standard for quantifi-cation of all the oligosaccharides.The calibration curve for DP3 showed a good linear regression(R2>0.9988).The limit of detection(LOD)and limit of quantification(LOQ)values were 0.05 mg/mL and 0.2 mg/mL,respectively.The recovery for DP3 was 87.32(±0.03)%in the three independent injections.To sum up,this proposed method is helpful for improving the quality control of polysaccharides from H.erinaceus as well as other materials.
