大孔吸附树脂法富集纯化黄芪多糖的工艺优化及评价研究  

Process optimization and evaluation of enrichment and purification of Astragalus polysaccharides by macroporous adsorption resin

作　　者：何海 赵沙沙 张小荣 崔治家 邵晶 HE Hai;ZHAO Shasha;ZHANG Xiaorong;CUI Zhijia;SHAO Jing(College of Pharmacy,Gansu University of Chinese Medicine,Lanzhou Gansu 730000,China;Northwest Collaborative Innovation Center for Traditional Chinese Medicine Co-constructed by Gansu Province&MOE of PRC,Lanzhou Gansu 730000,China;Key Laboratory of Pharmacology and Toxicology of Traditional Chinese Medicine in Gansu Province,Gansu University of Chinese Medicine,Lanzhou Gansu 730000,China;Research Center of Traditional Chinese Medicine Pharmaceutical Technology and Engineering in Gansu Province,Lanzhou Gansu 730000,China)

机构地区：[1]甘肃中医药大学药学院,甘肃兰州730000 [2]西北中藏药省部共建协同创新中心,甘肃兰州730000 [3]甘肃省中药药理与毒理学重点实验室,甘肃兰州730000 [4]甘肃省中药制药工艺工程研究中心,甘肃兰州730000

出　　处：《中兽医医药杂志》2022年第5期1-6,共6页Journal of Traditional Chinese Veterinary Medicine

基　　金：甘肃省自然科学基金项目(21JR11RA136);甘肃省自然科学基金项目(17JR5RA163);甘肃省中药制药工艺工程研究中心开放课题(ZYGY202004);兰州市城关区科技计划项目(2020-2-2-2);甘肃省教育厅双一流重大科研项目(GSSYLXM-05);中医药公共卫生服务补助专项子课题(2305191901)。

摘　　要：以研究黄芪多糖标准提取物为目的,筛选优化其大孔吸附树脂纯化富集的工艺方法与工艺参数,并进行评价分析。以吸附量、吸附率、解析率为指标,考察4种型号大孔吸附树脂(AB-8、D-101、NKA-9、X-5)对黄芪多糖的纯化富集效果,并进行相关工艺参数优化;采用改良差示苯酚硫酸法测定多糖含量;采用IR法、GC-MS法、体外总抗氧化试验对纯化前后的黄芪多糖进行化学特征分析与抗氧化活性评价。确定的纯化工艺为:NKA-9大孔树脂50 g湿法装柱(2 cm×50 cm),取质量浓度为3.0 mg/mL的黄芪粗多糖溶液250 mL,以2.0 mL/min流速循环上样2次,动态吸附后再静态吸附24 h,后以50%丙酮0.8 mL/min进行洗脱,洗脱液用量500 mL。验证试验表明,经本工艺纯化富集后,所得黄芪多糖纯度提高58.84%;IR光谱表现出更突出的多糖结构特征吸收峰。GC-MS法评价纯化后黄芪多糖的单糖组成为鼠李糖、阿拉伯糖、甘露糖、葡萄糖和半乳糖。与黄芪粗多糖相比较,NKA-9大孔吸附树脂纯化黄芪多糖总抗氧化活性显著增强。优化的黄芪多糖大孔树脂纯化工艺稳定可行,重复性较好,使多糖得到较好的纯化富集,为黄芪多糖标准提取物的制备及原料药研制提供了研究基础。To study the standard extract of Astragalus polysaccharides,screen and optimize the technological method and parameters of macroporous adsorption resin purification and enrichment,and carry out evaluation and analysis.Using adsorption capacity,adsorption rate and resolution rate as indicators,investigating the purification and enrichment effects of four types of macroporous adsorption resins(AB-8,d-101,NKA-9 and X-5)on Astragalus polysaccharides and optimizing the relevant process parameters;improved phenol sulfuric acid method were used to determine the content of polysaccharide;Astragalus polysaccharides before and after purification were evaluated by IR method,GC-MS method and total antioxidant test in vitro.The determined purification process was that NKA-9 macroporous resin 50 g wet column installation(2 cm×50 cm),250 mL astragalus crude polysaccharide solution with concentration of 3.0 mg·mL^(-1),circulate the sample twice at the flow rate of 2.0 mg·mL^(-1),static adsorption for 24 h after dynamic adsorption,then eluted with 50%acetone 0.8 mg·mL^(-1),and the amount of eluent was 500 mL.The validation experiment showed that the purity of Astragalus polysaccharide was increased by 58.84%after purification and enrichment,IR spectra showed more prominent structural characteristic absorption peak of polysaccharides.The monosaccharide composition of purified Astragalus polysaccharides was rhamnose,arabinose,mannose,glucose and galactose by GC-MS analysis.Compared with crude Astragalus polysaccharides,the total antioxidant activity of Astragalus polysaccharides purified by NKA-9 macroporous adsorption resin was significantly enhanced.The optimized HPS macroporous resin purification process is stable and feasible with good repeatability,so that the polysaccharides can be well purified and enriched,which provides a research basis for the formation of Astragalus polysaccharides standard extract and its becoming a scientific bulk drug of preparation.

关 键 词：黄芪多糖 大孔吸附树脂 富集纯化 工艺优化 抗氧化活性 

分 类 号：S853.72[农业科学—临床兽医学]