炙黄芪多糖的酶解产物指纹图谱构建与化学模式识别分析  

Fingerprints and chemical pattern recognition of enzyme hydrolysates of honey-processed Astragali Radix polysaccharides

作　　者：李亚栋 赵颖 尚奥丽 杨若泓 陈君 LI Ya-dong;ZHAO Ying;SHANG Ao-li;YANG Ruo-hong;CHEN Jun(School of Traditional Chinese Medicine,China Pharmaceutical University,Nanjing 210009)

机构地区：[1]中国药科大学中药学院,南京210009

出　　处：《中南药学》2024年第11期2994-2999,共6页Central South Pharmacy

基　　金：国家重点研发计划(No.2022YFC3501604)。

摘　　要：目的构建炙黄芪多糖酶解产物指纹图谱,推动其质量标准的提升。方法采用水提醇沉法制备炙黄芪多糖,然后用β-D-半乳糖苷酶进行水解,采用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF-MS)对水解产物进行鉴定,并运用亲水色谱-高效液相色谱-蒸发光散射检测器(HILIC-HPLC-ELSD)构建水解产物指纹图谱,结合相似度评价、聚类分析(HCA)和正交偏最小二乘判别分析(OPLS-DA)比较生黄芪、炙黄芪的差异。结果炙黄芪多糖水解产物经鉴定主要为葡萄糖(Glc)和聚合度(DP)为2~8的六碳糖寡糖片段,炙黄芪多糖中不存在游离的β-D-半乳糖基,还原端存在与半乳糖以β-D-糖苷键连接的游离葡萄糖基;炙黄芪样品的相似度为0.939~0.995,而生黄芪样品与炙黄芪多糖的相似度为0.855~0.923,HCA和OPLS-DA模型可将生黄芪、炙黄芪明显区分,Glc、DP2和DP3是两者水解产物的主要差异糖片段。结论本研究建立的炙黄芪多糖酶解产物指纹图谱,体现了多糖结构特征且专属性良好,可为炙黄芪的质量控制提供参考。Objective To establish the fingerprints of enzyme hydrolysates of saccharide mapping of honey-processed Astragali Radix(HAR)to improve the standard of HAR.Methods β-D-galactosidase was used to hydrolyze HAR polysaccharides(HAPS),and ultra-performance liquid chromatography and time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was used to identify the HAPS hydrolysates.Hydrophilic interaction-high performance liquid chromatography-evaporative light scattering detector(HILIC-HPLC-ELSD)was used to establish the fingerprints of HAPS hydrolysates.The similarity was evaluated,hierarchical cluster analysis(HCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were conducted compare the differences between HAR and Astragali Radix(AR).Results The hydrolysates were mainly composed of glucose(Glc)and oligosaccharides with degree of polymerization(DP)of 2~8.No free β-D-galactose was found in HAPS,and there was free glucose connected with galactose by β-D-glycosidic bond in the reducing end.The similarities of HAPS samples ranged from 0.939 to 0.995,while the similarities of HAPS samples and Astragalus polysaccharides(APS)samples ranged from 0.855 to 0.923.HCA and the established OPLS-DA model could distinguish HAPS samples from APS.Glc,DP2 and DP3 were the main sugar fragments in the hydrolysates of HAPS and APS.Conclusion The fingerprints of enzymatic hydrolysates of HAPS established in this study reflect the structural characteristics of polysaccharides with good specificity,which provides reference for the quality control of HAR.

关 键 词：黄芪多糖 炙黄芪 酶解 指纹图谱 质量评价 

分 类 号：R283.6[医药卫生—中药学] R284.1[医药卫生—中医学]