基于非对称场流分离技术的山药多糖表征及其抗氧化活性研究

doi:10.3969/j.issn.1000-1565.2026.01.006

摘要/Abstract

摘要： 山药多糖具有重要的生物活性,本文采用超声辅助热水提取法提取了棒山药和小白嘴山药多糖,通过酶解法去除蛋白质,采用非对称场流分离技术在线联用多角度光散射检测器和示差折光检测器(AF4-MALS-dRI)、傅里叶变换红外光谱和刚果红实验对山药多糖的结构进行了表征,对比了Berry和Debye 2种光散射函数数学模型对山药多糖摩尔质量和回转半径表征结果准确性的影响,研究了山药多糖的抗氧化活性.结果证明2种山药多糖均具有β-D-吡喃环结构,其中棒山药多糖具有相对松散的三螺旋结构.在2 mg/mL质量浓度下,棒山药多糖的OH和DPPH自由基清除率分别为(93.9±0.29)%和(88.2±0.58)%,高于小白嘴山药的抗氧化活性.结果表明相对松散的三螺旋结构有利于山药多糖的抗氧化活性.

关键词: 非对称场流分离, 山药多糖, 抗氧化活性

Abstract: Yam polysaccharides(YPs)have important biological activities. The polysaccharides were extracted from Bang yam and Xiaobaizui yam by the ultrasound-assisted hot water extraction method. The proteins were removed from the crude yam polysaccharide by enzymatic hydrolysis. The structure of YPs was characterized by using asymmetrical flow field-flow fractionation coupled online with multiangle light scattering detector and differential refractive index detector(AF4-MALS-dRI), Fourier transform infrared spectroscopy, and Congo red experiment. The effect of Berry and Debye models on the accuracy of the characterization results of YPs(i.e., molar mass and radius of gyration)was investigated. Furthermore, the antioxidant activity of YPs was studied. The results suggested that the two YPs had a β-D-pyran ring structure. The Bang YP(BYP)had a relatively loose triple helix structure. At a concentration of 2 mg/mL,- 引用格式:张玲,彭芯钰,章瑞环,等.基于Winkler地基梁理论的拼宽公路新老路基差异沉降分析［J］.河北大学学报(自然科学版),2026,46(1):1-12.引用格式:张孜寒,刘晓莹,尹亭惠,等.基于非对称场流分离技术的山药多糖表征及其抗氧化活性研究［J］.河北大学学报(自然科学版),2026,46(1):48-55.DOI:10.3969/j.issn.1000-1565.2026.01.006基于非对称场流分离技术的山药多糖表征及其抗氧化活性研究张孜寒,刘晓莹,尹亭惠,郭子瑶,窦海洋(河北大学基础医学院,河北省炎性自身免疫性疾病发病机制及防治重点实验室,河北保定 071000)摘要:山药多糖具有重要的生物活性,本文采用超声辅助热水提取法提取了棒山药和小白嘴山药多糖,通过酶解法去除蛋白质,采用非对称场流分离技术在线联用多角度光散射检测器和示差折光检测器(AF4-MALS-dRI)、傅里叶变换红外光谱和刚果红实验对山药多糖的结构进行了表征,对比了Berry和Debye 2种光散射函数数学模型对山药多糖摩尔质量和回转半径表征结果准确性的影响,研究了山药多糖的抗氧化活性.结果证明2种山药多糖均具有β-D-吡喃环结构,其中棒山药多糖具有相对松散的三螺旋结构.在2 mg/mL质量浓度下,棒山药多糖的OH和DPPH自由基清除率分别为(93.9±0.29)%和(88.2±0.58)%,高于小白嘴山药的抗氧化活性.结果表明相对松散的三螺旋结构有利于山药多糖的抗氧化活性.关键词:非对称场流分离;山药多糖;抗氧化活性中图分类号:O657 文献标志码:A 文章编号:1000-1565(2026)01-0048-08DOI:10.3969/j.issn.1000-1565.2026.01.006Characterization of yam polysaccharide by asymmetrical flow field-flowfractionation and study on its antioxidant activityZHANG Zihan, LIU Xiaoying, YIN Tinghui, GUO Ziyao, DOU Haiyang(Key Laboratory of Pathogenesis Mechanism and Control of Inflammatory-Autoimmune Disease ofHebei Province, School of Basic Medical Sciences, Hebei University, Baoding 071000, China)Abstract: Yam polysaccharides(YPs)have important biological activities. The polysaccharides were extracted from Bang yam and Xiaobaizui yam by the ultrasound-assisted hot water extraction method. The proteins were removed from the crude yam polysaccharide by enzymatic hydrolysis. The structure of YPs was characterized by using asymmetrical flow field-flow fractionation coupled online with multiangle light scattering detector and differential refractive index detector(AF4-MALS-dRI), Fourier transform infrared spectroscopy, and Congo red experiment. The effect of Berry and Debye models on the accuracy of the characterization results of YPs(i.e., molar mass and radius of gyration)was investigated. Furthermore, the antioxidant activity of YPs was studied. The results suggested that the two YPs had a β-D-pyran ring structure. The Bang YP(BYP)had a relatively loose triple helix structure. At a concentration of 2 mg/mL,- 收稿日期:2025-01-25;修回日期:2025-04-30 基金项目:河北省自然科学基金项目(H2024201027);大学生创新创业训练计划项目(DC2024288;DC2005546) 第一作者:张孜寒(2002—),女,河北大学在读硕士研究生,主要从事场流分离方向研究. E-mail: zhangzihan62024@163.com 通信作者:窦海洋(1983—),男,河北大学副教授,博士,主要从事场流分离方向研究.E-mail: douhaiyang-1984@163.com 第1期张孜寒等:基于非对称场流分离技术的山药多糖表征及其抗氧化活性研究河北大学学报(自然科学版) 第46卷the scavenging rates of BYP for OH and DPPH free radicals were(93.9±0.29)% and(88.2±0.58)%, respectively, which were higher than those of Xiaobaizui YP(XYP). The results revealed that the relatively loose triple helix structure could be beneficial to the antioxidant activity of YP.

Key words: asymmetrical flow field-flow fractionation, yam polysaccharide, antioxidant activity

中图分类号:

O657

张孜寒,刘晓莹,尹亭惠,郭子瑶,窦海洋. 基于非对称场流分离技术的山药多糖表征及其抗氧化活性研究[J]. 河北大学学报(自然科学版), 2026, 46(1): 48-55.

ZHANG Zihan, LIU Xiaoying, YIN Tinghui, GUO Ziyao, DOU Haiyang. Characterization of yam polysaccharide by asymmetrical flow field-flow fractionation and study on its antioxidant activity[J]. Journal of Hebei University(Natural Science Edition), 2026, 46(1): 48-55.

参考文献

[1] LI Y, JI S Y, XU T, et al.Chinese yam(Dioscorea): Nutritional value, beneficial effects, and food and pharmaceutical applications[J].Trends Food Sci Technol, 2023, 134: 29-40. DOI: 10.1016/j.tifs.2023.01.021.
[2] ZHANG L, WANG S R, ZHANG W M, et al. Prospects of yam(Dioscorea)polysaccharides: Structural features, bioactivities and applications[J]. Food Chem, 2024, 446: 138897. DOI: 10.1016/j.foodchem.2024.138897.
[3] CHANG H Y, TONG X Y, YANG H Q, et al. Chinese yam(Dioscorea opposita)and its bioactive compounds: The beneficial effects on gut microbiota and gut health[J]. Curr Opin Food Sci, 2024, 55: 101121. DOI: 10.1016/j.cofs.2023.101121.
[4] OUYANG J, WANG F, LI W T, et al. Structure characterization of polysaccharide from Chinese yam(Dioscorea opposite Thunb.)and its growth-promoting effects on streptococcus thermophilus[J]. Foods, 2021, 10(11): 2698. DOI: 10.3390/foods10112698.
[5] ZHOU S Y, HUANG G L, CHEN G Y. Extraction, structural analysis, derivatization and antioxidant activity of polysaccharide from Chinese yam[J]. Food Chem, 2021, 361: 130089. DOI: 10.1016/j.foodchem.2021.130089.
[6] BAI C L, CHEN R I, CHEN Y B, et al. Plant polysaccharides extracted by high pressure: A review on yields, physicochemical, structure properties, and bioactivities[J]. Int J Biol Macromol, 2024, 263: 129939. DOI: 10.1016/j.ijbiomac.2024.129939.
[7] FENG X J, GUO M Z, LI J Y, et al. The structural characterization of a novel Chinese yam polysaccharide and its hypolipidemic activity in HFD-induced obese C57BL/6J mice[J]. Int J Biol Macromol, 2024, 265: 130521. DOI: 10.1016/j.ijbiomac.2024.130521.
[8] LI Z D, XIAO W H, XIE J H, et al. Isolation, characterization and antioxidant activity of yam polysaccharides[J]. Foods, 2022, 11(6): 800. DOI: 10.3390/foods11060800.
[9] JUROWSKI K, NOGA M, KOBYLARZ D, et al. Multimodal imaging using raman spectroscopy and FTIR in a single analytical instrument with a microscope(Infrared Raman Microscopy AIRsight, Shimadzu): Opportunities and applications[J]. Int J Mol Sci, 2024, 25(13): 6884. DOI: 10.3390/ijms25136884.
[10] CHEAH Y T, CHAN D J C. A methodological review on the characterization of microalgal biofilm and its extracellular polymeric substances[J]. J Appl Microbiol, 2022, 132(5): 3490-3514. DOI: 10.1111/jam.15455.
[11] WANG Z F, YOU Y L, Li F F, et al. Research progress of NMR in natural product quantification[J]. Molecules, 2021, 26(20): 6308. DOI: 10.3390/molecules26206308.
[12] CHEN X, ZHANG W H, DOU Y W, et al. Applications of asymmetrical flow field-flow fractionation for separation and characterization of polysaccharides: A review[J]. J Chromatogr A, 2021, 1635: 461726. DOI: 10.1016/j.chroma.2020.461726.
[13] 张晨婧,张西瑞,郭子瑶,等.大豆7S球蛋白-大枣多糖复合物的制备、表征及抗原性[J].河北大学学报(自然科学版), 2024, 44(3): 254-260. DOI: 10.3969/j.issn.1000-1565.2024.03.004.
[14] VENTOURI I K, ASTEFANEI A, KAAL E R, et al. Asymmetrical flow field-flow fractionation to probe the dynamic association equilibria of β-D-galactosidase[J]. J Chromatogr A, 2021, 1635: 461719. DOI: 10.1016/j.chroma.2020.461719.
[15] IBIETA G, BUSTOS A-S, ORTIZ-SEMPERTEGUI J, et al. Molecular characterization of a galactomannan extracted from Tara(Caesalpinia spinosa)seeds[J]. Sci Rep, 2023, 13(1): 21893. DOI: 10.1038/s41598-023-49149-3.
[16] GUO Y X, ZHANG W H, SUN Y S, et al. Asymmetrical flow field-flow fractionation combined with liquid chromatography enables rapid, quantitative, and structurally informative detection of resistant starch[J]. Microchem J, 2022, 181: 107787. DOI: 10.1016/j.microc.2022.107787.
[17] LIN B B, WANG S S, ZHOU A Q, et al. Ultrasound-assisted enzyme extraction and properties of Shatian pomelo peel polysaccharide[J]. Ultrason Sonochem, 2023, 98: 106507. DOI: 10.1016/j.ultsonch.2023.106507.
[18] AL-WRAIKAT M, LIU Y, WU L M, et al. Structural characterization of degraded Lycium barbarum L. Leaves' polysaccharide using ascorbic acid and hydrogen peroxide[J]. Polymers,2022,14(7):1404. DOI: 10.3390/polym14071404.
[19] HUO N L, AMEER K, WU Z G, et al. Preparation, characterization, structural analysis and antioxidant activities of phosphorylated polysaccharide from Sanchi(Panax notoginseng)flower[J]. J Food Sci Technol, 2022, 59(12): 4603-4614. DOI: 10.1007/s13197-022-05539-5.
[20] CHEN N-D, CHEN N F, LI J, et al. Similarity evaluation of different origins and species of dendrobiums by GC-MS and FTIR analysis of polysaccharides[J]. Int J Anal Chem, 2015, 2015(1): 713410. DOI: 10.1155/2015/713410.
[21] ANDERSSON M, WITTGREN B, WAHLUND K G. Accuracy in multiangle light scattering measurements for molar mass and radius estimations: Model calculations and experiments[J]. Anal Chem, 2003, 75(16): 4279-4291. DOI: 10.1021/ac030128+.
[22] JUNA S, WILLIAMS P A, Davies S. Determination of molecular mass distribution of amylopectin using asymmetrical flow field-flow fractionation[J]. Carbohydr Polym, 2011, 83(3): 1384-1396. DOI: 10.1016/j.carbpol.2010.09.045.
[23] ZHANG W H, SHEN S G, SONG T G, et al. Insights into the structure and conformation of potato resistant starch(type 2)using asymmetrical flow field-flow fractionation coupled with multiple detectors[J]. Food Chem, 2021, 349: 129168. DOI: 10.1016/j.foodchem.2021.129168.
[24] GRIMM A, KRÜGER E, BURCHARD W. Solution properties of β-D-(1, 3)(1, 4)-glucan isolated from beer[J]. Carbohydr Polym, 1995, 27(3): 205-214. DOI: 10.1016/0144-8617(95)00056-D.
[25] 张汇,聂少平,艾连中,等. 灵芝多糖的结构及其表征方法研究进展[J]. 中国食品学报, 2020, 20(1): 290-301. DOI: 10.16429/j.1009-7848.2020.01.038. (