不同取代度蕨麻硒多糖的制备、表征及体外抗氧化活性
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摘要
硒多糖具有抗金属中毒、抗氧化、抗病毒等多种作用,其结构与活性密切相关。本文以从我国传统藏药蕨麻中提取的蕨麻多糖为前体物,通过单因素和响应面设计来优化工艺并调控条件制备出不同取代度的蕨麻硒多糖,使用尺寸排除色谱-激光光散射联用仪、紫外光谱仪、红外光谱仪、热重分析仪、扫描电镜对产物结构进行表征,最后评估了不同取代度蕨麻硒多糖的体外抗氧化活性。结果表明:蕨麻硒多糖的最佳硒化时间为134.84 min,温度78.39℃,催化剂量49.37 mg,投料比PAP∶H_2SeO_3=1∶0.81,此条件下产物的硒含量可达到8518.81μg/g;制备出的五种取代度的蕨麻硒多糖结构中均含有Se=O键和Se-O键,即实现了蕨麻多糖的硒化;取代度越高的蕨麻硒多糖的分子量越小、热稳定性越低、空间结构越松散,蕨麻硒多糖对O~-_2·、·OH和DPPH·的清除能力明显且随硒取代度的增加而增强。
Selenium polysaccharide has multiple functions of anti-metal poisoning,anti-oxidation,anti-virus etc. Its activity is closely related to its structure. In this paper,the Potentilla anserina L. polysaccharide extracted from traditional Tibetan medicine potentilla was used as a precursor. Optimizing the preparation process and regulated the conditions by single factor and response surface design to prepare different substitution degrees of Potentilla anserina L. polysaccharides. The size exclusion chromatography-laser light scattering spectroscopy,UV spectrometer,infrared spectrometer,thermogravimetric analyzer,and scanning electron microscopy were used to characterize the structure of the product and evaluated the in vitro antioxidant activity of different substitution degrees of selenium Potentilla anserina L. polysaccharide in the end. The results showed that the optimum reaction time of selenium Potentilla anserina L. polysaccharide was 134.84 min,temperature 78.39 ℃,catalyst 49.37 mg and wight ratio PAP∶H_2SeO_3=1∶0.81. Under these conditions,the selenium contents of the product was 8518.81 μg/g. There existed Se=O and Se-O bonds in selenium Potentilla anserina L. polysaccharide with five degrees of substitution,meaning the selenization of the Potentilla anserina L.polysaccharide was successful. The higher degree of substitution of selenium Potentilla anserina L. polysaccharide,the smaller the molecular weight,the lower the thermal stability,and the looser the spatial structure. The selenium Potentilla anserina L. polysaccharide had significant scavenging ability to O~-_2·,·OH and DPPH·,and had positive correlation to the substitution degree of selenium.
Selenium polysaccharide has multiple functions of anti-metal poisoning,anti-oxidation,anti-virus etc. Its activity is closely related to its structure. In this paper,the Potentilla anserina L. polysaccharide extracted from traditional Tibetan medicine potentilla was used as a precursor. Optimizing the preparation process and regulated the conditions by single factor and response surface design to prepare different substitution degrees of Potentilla anserina L. polysaccharides. The size exclusion chromatography-laser light scattering spectroscopy,UV spectrometer,infrared spectrometer,thermogravimetric analyzer,and scanning electron microscopy were used to characterize the structure of the product and evaluated the in vitro antioxidant activity of different substitution degrees of selenium Potentilla anserina L. polysaccharide in the end. The results showed that the optimum reaction time of selenium Potentilla anserina L. polysaccharide was 134.84 min,temperature 78.39 ℃,catalyst 49.37 mg and wight ratio PAP∶H_2SeO_3=1∶0.81. Under these conditions,the selenium contents of the product was 8518.81 μg/g. There existed Se=O and Se-O bonds in selenium Potentilla anserina L. polysaccharide with five degrees of substitution,meaning the selenization of the Potentilla anserina L.polysaccharide was successful. The higher degree of substitution of selenium Potentilla anserina L. polysaccharide,the smaller the molecular weight,the lower the thermal stability,and the looser the spatial structure. The selenium Potentilla anserina L. polysaccharide had significant scavenging ability to O~-_2·,·OH and DPPH·,and had positive correlation to the substitution degree of selenium.
引文
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[23]李珺,钟耀广,刘长江.香菇多糖的纯化及电镜分析[J].山西农业科学,2010,38(3):6-9.
[2]梁欢,黄进,王丽,等.药用植物硒多糖的研究进展[J].中国中药杂志,2018(15).
[3]Surhio M M,Wang Y,Xu P,et al.Antihyperlipidemic and hepatoprotective properties of selenium modified polysaccharide from Lachnum sp[J].International Journal of Biological Macromolecules,2017,99.
[4]Ferreira I C,Heleno S A,Reis F S,et al.Chemical features of Ganoderma polysaccharides with antioxidant,antitumor and antimicrobial activities[J].Phytochemistry,2015,114:38-55.
[5]魏慧.蕨麻多糖对镉致免疫抑制小鼠的免疫调节机理研究[D].兰州:兰州大学,2016.
[6]葛肖健,张永亮,师超峰,等.蕨麻多糖对缺氧损伤人脐静脉内皮细胞的保护作用及机制[J].山东医药,2017,57(5):8-11.
[7]闵光涛,冯颖,闵光宁,等.蕨麻提取物保护小鼠四氯化碳急性肝损伤的作用机制[J].兰州大学学报(医学版),2012,38(1):49-52.
[8]Zhao B,Zhang J,Yao J,et al.Selenylation modification can enhance antioxidant activity of Potentilla anserine L.polysaccharide[J].International Journal of Biological Macromolecules,2013,58(2):320-328.
[9]柴刚,魏洪敏,王俊杰,等.原子荧光法测定花椰菜、青花菜中的硒含量[J].农产品质量与安全,2018(1):80-83.
[10]Ding W,Zhou J,Zeng Y,et al.Preparation of oxidized sodium alginate with different molecular weights and its application for crosslinking collagen fiber[J].Carbohydrate Polymers,2017,157:1650-1656.
[11]封燕,贡小辉,韦德群,等.金蝉花多糖的抗氧化活性及结构分析[J].食品科学,2016,37(12):19-24.
[12]秦利鸿,曹建波,易伟松.绿茶多糖的扫描电镜制样新方法及原子力显微镜观察[J].电子显微学报,2009,28(2):162-167.
[13]陈晋明,冯翠萍.白灵菇多糖抗氧化活性研究[J].食品科技,2015,40(2):239-242.
[14]贾之慎,邬建敏.比色法测定Fenton法产生的羟基自由基[J].生物化学与生物物理进展,1996,23(2):184-186.
[15]宋佳敏,王鸿飞,孙朦,等.响应面法优化金蝉花多糖提取工艺及抗氧化活性分析[J].食品科学,2018,39(4):275-281.
[16]白婕,郭凯,沈银梅.Box-Behnken响应面法优化富硒平菇柄多糖提取工艺研究[J].经济林研究,2017,35(2):121-126.
[17]陈兵兵,王振斌.葛根多糖的基本理化特性研究[J].食品研究与开发,2016,37(15):10-13.
[18]Wang D,Sun S Q,Wu W Z,et al.Characterization of a water-soluble polysaccharide from Boletus edulis,and its antitumor and immunomodulatory activities on renal cancer in mice[J].Carbohydr Polym,2014,105(1):127-134.
[19]王占一,戴博,张立华,等.石榴皮多糖硒酸酯制备工艺参数优化及其结构分析[J].食品科学,2016,37(10):40-46.
[20]葛明明,胡北,孙丽娜,等.硒化蒲公英多糖的制备工艺及硒含量测定的研究[J].食品工业科技,2014,35(8):276-280.
[21]赵玉苹.海藻硒多糖的合成及生物活性研究[D].广州:暨南大学,2006.
[22]高义霞,袁毅君,周向军,等.罗望子多糖硒酸酯的制备及表征[J].药物分析杂志,2012(7):1222-1226.
[23]李珺,钟耀广,刘长江.香菇多糖的纯化及电镜分析[J].山西农业科学,2010,38(3):6-9.