[关键词]
[摘要]
目的 比较研究黄芪蜜炙后多糖、单糖、寡糖及非糖类小分子成分的整体变化。方法 结合糖组学和代谢组学,运用超高效液相-二极管阵列检测器(ultra high performance liquid-photodiode array detector,UPLC-PDA)、高效液相-蒸发光散射检测器(ultra high performance liquid-evaporative light scattering detector,HPLC-ELSD)、高效凝胶渗透色谱-蒸发光散射检测器(high performance gel permeation chromatography-evaporative light scattering detector,HPGPC-ELSD)和超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(ultra high performance liquid chromatography-quadrupole/electrostatic field orbit-trap high resolution mass spectrometr,UHPLC-Q-Orbitrap HRMS)等对黄芪和蜜炙黄芪的多糖、单糖、寡糖以及非糖小分子组分进行分析。结果 黄芪和蜜炙黄芪中均可以找到3种相对分子质量的多糖,但3种多糖的重均相对分子质量、相对分子质量分布以及3种多糖各自的占比存在差异。黄芪和蜜炙黄芪多糖均含有甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖、岩藻糖7种单糖,但两者物质的量比稍有不同(黄芪vs蜜炙黄芪=0.70∶0.20∶0.85∶5.67∶0.44∶2.04∶0.32 vs 0.79∶0.15∶0.73∶7.10∶0.31∶1.71∶0.35)。在黄芪和蜜炙黄芪中均能检测到寡糖蔗糖,蜜炙后蔗糖含量显著下降(P<0.01)。黄芪和蜜炙黄芪均含有游离单糖果糖、甘露糖、葡萄糖、半乳糖醛酸和岩藻糖,蜜炙黄芪中的果糖、甘露糖、葡萄糖含量均显著高于黄芪(P<0.05、0.01),而半乳糖醛酸的含量显著低于黄芪(P<0.05),岩藻糖含量在黄芪蜜炙后无显著变化。从黄芪和蜜炙黄芪中共筛选出20个差异性非糖小分子成分,主要为皂苷类和黄酮类成分。代表性成分定量分析表明,与黄芪相比,蜜炙黄芪中的黄酮类成分毛蕊异黄酮葡萄糖苷、芒柄花素、芒柄花苷的含量显著降低(P<0.05、0.01),皂苷类成分黄芪甲苷含量显著增加(P<0.01)。呋喃醛衍生物5-羟甲基糠醛在黄芪中未检测到,在蜜炙黄芪中的含量显著升高(P<0.001)。结论 黄芪和蜜炙黄芪的糖类组分与非糖类组分存在显著差异,这可能是造成黄芪蜜炙后补中益气功效增强的主要原因。
[Key word]
[Abstract]
Objective The overall variations on polysaccharides, monosaccharides, oligosaccharides and other chemical components were studied when Huangqi (Astragali Radix,AR) processed by honey. Methods Merging glycomics and metabolomics methods were used to analyze the polysaccharides, monosaccharides, oligosaccharides and other chemical components of Astragali Radix and honey processed Astragali Radix (HAR) by using ultra high performance liquid-photodiode array detector (UPLC-PDA), ultra high performance liquid-evaporative light scattering detector (HPLC-ELSD), high performance gel permeation chromatography-evaporative light scattering detector (HPGPC-ELSD) and ultra high performance liquid chromatography-quadrupole/electrostatic field orbit-trap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Result Three kinds of polysaccharides found in both AR and HAR, however, the relative molecular weight, molecular weight distribution and proportion of the three polysaccharides in AR and HAR were different. Both AR and HAR polysaccharides contained mannose, rhamnose, galacturonic acid, glucose, galactose, arabinose, and fucose, but their molar ratios were slightly different (AR vs HAR = 0.70∶0.20∶0.85∶5.67∶0.44∶2.04∶0.32 vs 0.79∶0.15∶0.73∶7.10∶0.31∶1.71∶0.35). Oligosaccharides sucrose could be detected in both AR and HAR, but the content was decreased significantly after honey treatment (P<0.01). Free monosaccharides like fructose, mannose, glucose, galacturonic acid and fucose were also detected both in AR and HAR, and the contents of fructose, mannose and glucose were significantly increased after honey treatment (P<0.05, 0.01). At the same time, the content of galacturonic acid (P<0.05) was significantly decreased. There was no significant change in fucose after Astragali Radix honey processing. For other chemical components, a total of 20 markers were screened when AR processed by honey, which were mainly saponins and flavonoids. The quantitative analysis results showed that flavonoid components like calycosin-7-O-β-D-glucoside, formononetin, and ononin were significantly decreased (P<0.05, 0.01), while the content of astragaloside IV (P<0.01) was significantly increased. As the furfural derivative, 5-hydroxymethylfurfural (P<0.001) was not detected in AR, but significantly increased in HAR. Conclusion There are significant differences in saccharides ingredients and other chemical components between AR and HAR, which may be the main reason for the enhanced effect of HAR in tonifying middle and replenishing qi.
[中图分类号]
R284.1
[基金项目]
国家自然科学基金面上项目(82074022);国家自然科学基金联合基金重点项目(U21A20410);科技部“中医药现代化研究”重点研发计划项目子课题(2019YFC1710800);山西省应用基础研究项目(201801D221436);山西省教育厅科技创新计划项目(2019L0720);山西中医药大学科技创新计划项目(2022PY-YH-13);山西省中医药管理局科研项目(2022ZYYC265,2023ZYYA033)