粉红女士苹果多糖分离纯化、结构表征及抗氧化活性研究
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摘要
提取粉红女士苹果多糖半纯品并纯化,比较苹果多糖半纯品和纯品的表征性结构和抗氧化活性差异。以粉红女士苹果为试材,通过水提醇沉得多糖半纯品(PPS),用DEAE-52纤维素柱进一步分离纯化收集纯品组分,苯酚硫酸法、气相色谱法、高效凝胶渗透色谱法分别测定各多糖组分的含量、单糖组成和分子质量,进一步用傅里叶变换红外光谱和差示扫描量热法表征各多糖组分。结果表明:PPS上样量200 mg,以0.1,0.2 mol/L Na Cl水溶液依次在DEAE-52柱上洗脱得两种纯品组分(PPS-1和PPS-2)。PPS,PPS-1,PPS-2多糖含量分别为67.32%,89.74%,90.88%,重均分子质量分别为111.28,52.79,258.39 ku,单糖组成相同,其中PPS中半乳糖醛酸、PPS-1中阿拉伯糖和半乳糖醛酸、PPS-2中半乳糖和半乳糖醛酸分别占51.6%,69.9%和81.3%。PPS,PPS-1,PPS-2均具有植物多糖特征吸收峰,三者的热学特性与化学组成吻合。抗氧化试验表明,PPS还原力和清除DPPH·,ABTS+·能力高于PPS-1和PPS-2,三者清除·OH能力无显著差异。PPS较PPS-1、PPS-2制备简单,抗氧化活性高,可作为粉红女士苹果的高附加值产品运用到功能食品、医学和其它相关领域。
Studied with pink lady apple polysaccharide, an investigation of the differentiation of characterization and antioxidant activity of half pure and pure product were carried out in this study. Firstly,half pure product of pink lady apple polysaccharide(PPS) was gained with water extract-alcohol precipitation; secondly, the PPS was further purified by chromatography of Cellulose DEAE-52 to get pure fractions. Thirdly, the content of the polysaccharide components,monosaccharide composition and molecular weight were respectively determined by phenol sulfuric acid method, gas chromatography, high efficiency and gel permeation chromatography(HPGPC), and polysaccharide components were characterized using Fourier transform infrared spectroscopy(FTIR) and differential scanning calorimetry(DSC). The conclusions are as follows: 1) Two kinds of single products(PPS-1 and PPS-2) were got when the sample amount of PPS was200 mg and the DEAE-52 column was eluted with 0.1, 0.2 mol/L Na Cl aqueous solution in turn; 2) the polysaccharide content of PPS, PPS-1 and PPS-2 were 67.32%, 89.74% and 90.88% respectively, the molecular weight of PPS, PPS-1 and PPS-2 were 111.28, 52.79, 258.39 ku respectively. Moreover, the monosaccharide composition of PPS, PPS-1 and PPS-2 was same. But the mole percentage of galacturonic acid in PPS, arabinose and galacturonic acid in PPS-1,galactose and galacturonic acid in PPS-2 were 51.6%, 69.9% and 81.3%, respectively; 3) FTIR analysis showed that PPS, PPS-1 and PPS-2 had polysaccharide characteristic absorption peak. The thermal property of the three fractions were consistent with their chemical composition. Antioxidant test revealed that reducing power and removing DPPH·,ABTS+· ability of PPS was higher than PPS-1 and PPS-2 and there was no significant difference in removing ·OH. The purification conditions of pink lady apple were first time optimized and reproducible. The condition was suitable for efficient preparation of pure polysaccharides from pink lady apple. With the advantage of simple-prepared and high antioxidant activity of PPS, compared with PPS-1 and PPS-2, it could be used as the pink lady apple value-added products in functional food, medicine and other related fields. The data presented here provides not only rationale for utilization of pink lady apple in general, but also a base for further studies of industrial use of biochemicals in the new apple variety.It suggested that pink lady could be a new plant polysaccharides resources using Industrial production.
Studied with pink lady apple polysaccharide, an investigation of the differentiation of characterization and antioxidant activity of half pure and pure product were carried out in this study. Firstly,half pure product of pink lady apple polysaccharide(PPS) was gained with water extract-alcohol precipitation; secondly, the PPS was further purified by chromatography of Cellulose DEAE-52 to get pure fractions. Thirdly, the content of the polysaccharide components,monosaccharide composition and molecular weight were respectively determined by phenol sulfuric acid method, gas chromatography, high efficiency and gel permeation chromatography(HPGPC), and polysaccharide components were characterized using Fourier transform infrared spectroscopy(FTIR) and differential scanning calorimetry(DSC). The conclusions are as follows: 1) Two kinds of single products(PPS-1 and PPS-2) were got when the sample amount of PPS was200 mg and the DEAE-52 column was eluted with 0.1, 0.2 mol/L Na Cl aqueous solution in turn; 2) the polysaccharide content of PPS, PPS-1 and PPS-2 were 67.32%, 89.74% and 90.88% respectively, the molecular weight of PPS, PPS-1 and PPS-2 were 111.28, 52.79, 258.39 ku respectively. Moreover, the monosaccharide composition of PPS, PPS-1 and PPS-2 was same. But the mole percentage of galacturonic acid in PPS, arabinose and galacturonic acid in PPS-1,galactose and galacturonic acid in PPS-2 were 51.6%, 69.9% and 81.3%, respectively; 3) FTIR analysis showed that PPS, PPS-1 and PPS-2 had polysaccharide characteristic absorption peak. The thermal property of the three fractions were consistent with their chemical composition. Antioxidant test revealed that reducing power and removing DPPH·,ABTS+· ability of PPS was higher than PPS-1 and PPS-2 and there was no significant difference in removing ·OH. The purification conditions of pink lady apple were first time optimized and reproducible. The condition was suitable for efficient preparation of pure polysaccharides from pink lady apple. With the advantage of simple-prepared and high antioxidant activity of PPS, compared with PPS-1 and PPS-2, it could be used as the pink lady apple value-added products in functional food, medicine and other related fields. The data presented here provides not only rationale for utilization of pink lady apple in general, but also a base for further studies of industrial use of biochemicals in the new apple variety.It suggested that pink lady could be a new plant polysaccharides resources using Industrial production.
引文
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[14]DU Y Q,WANG J H.Structural analysis and antioxidant activities of polysaccharideisolated from Jinqian mushroom[J].International Journal of Biological Macromolecules,2014,64(2):63-68.
[15]JIA D,ZHANG J,LAN R,et al.A simple preparative method for isolation and purification of polysaccharides from mulberry(Morus alba L.)leaves[J].International Journal of Food Science&Technology,2013,48(6):1275-1281.
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[18]SHARMA R,AHUJA M.Thiolated pectin:synthesis,characterization andevaluation as a mucoadhesive polymer[J].Carbohydrate Polymers,2011,85(3):658-663.
[19]张淑娟,徐怀德,米林峰.光皮木果汁体外抗氧化活性研究[J].食品科学,2011,32(21):56-61.
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[22]ZHU Z Y,NIAN L,SI C L,et al.Structure and antitumor activity of a high-molecular-weight polysaccharide from cultured mycelium of Cordyceps gunnii[J].Carbohydrate Polymers,2012,88(3):1072-1076.
[23]加藤宏治,新田正已,原田政子,等.糖质的红外线吸收光谱[J].静冈大学农学部研究报告,1972,22:77-85.
[24]张颖,曾艳,张丽姣,等.蛹虫草菌糠多糖的分离纯化及结构组成分析[J].食品科学,2014,35(13):54-59.
[25]SHEN S,CHENG H R,LI XU,et al.Effects of extraction methods on antioxidant activities of polysaccharides from camellia seed cake[J].European Food Research and Technology,2014,238(6):1015-1021.
[26]LIU L,CAO J,HUANG J,et al.Extraction of pectins with different degrees of esterification from mulberry branch bark[J].Bioresource Technology,2010,101(9):3268-3273.
[27]IIJIMA M,NAKAMURA K,HATAKEYAMA T,et al.Phase transition of pectin with sorbed water[J].Carbohydrate Polymers,2000,41(1):101-106.
[28]KANG,DUCK-HEE R N.Oxidative stress,DNA damage,and breast cancer[J].AACN Advanced Critical Care,2002,13(4):540-549.
[29]G譈L覶IN I.Antioxidant and antiradical activities of L-carnitine[J].Life science,2006,78(8):803-811.
[30]CHEN G T,MA X M,LIU S G,et al.Isolation,purification and antioxidant activities of polysaccharides from Grifola frondosa[J].Carbohydrate Polymers,2012,89(1):61-66.
[31]WANG Y F,YANG Z W,WEI X L,et al.Antioxidant activities potential of tea polysaccharide fractions obtained by ultra filtration[J].International Journal of Biological Macromolecules,2012,50(3):558-564.
[32]杨江涛,杨娟,谢红,等.刺梨多糖粗品与纯品体外抗氧化作用[J].食品工业科技,2008,13(2):94-96.
[2]JAHANBIN K,GOHARI A R,MOINIC S,et al.Isolation,structural characterization and antioxidant activity of a new water-soluble polysaccharide from Acanthophyllum bracteatum roots[J].International Journal of Biological Macromolecules,2011,49(4):567-572.
[3]Schieber A,Hilt P,Streker P,et al.A new process for the combined recovery of pectin and phenolic compounds from apple pomace[J].Innnovative Food Sciernce&Emerging Technologies,2003,4(1):99-107.
[4]马惠玲,盛义保,张丽萍,等.苹果渣果胶多糖的分离纯化与抗氧化活性研究[J].农业工程学报,2008,24(1):218-222.
[5]马俊杰.苹果渣中水溶性多糖的提取及其性质研究[D].兰州:甘肃农业大学,2009.
[6]YANG X B,YANG S,GUO Y R,et al.Compositional characterisation of soluble apple polysaccharides,and their antioxidant and hepatoprotective effects on acute CCl4-caused liver damage in mice[J].Food Chemistry,2013,138(2/3):1256-1264.
[7]ZHOU C L,LIU W,KONG Q,et al.Isolation,characterisation and sulphation of soluble polysaccharides isolated from Cucurbita maxima[J].International Journal of Food Science and Technology,2014,49(2):508-514.
[8]XIE L Y,PENG W H,GAN B C,et al.Optimization of protein removal method and condition of polysaccharide from Phellinus linteus[J].Agricultural Science&Technology,2011,12(9):1249-1251.
[9]苏钰琦.苹果多糖的分离纯化与抗氧化活性深入研究[D].杨凌:西北农林科技大学,2008.
[10]ZHANG X H,LIU L N,LIN C W,et al.Isolation,structural characterization and antioxidant activity of a neutral polysaccharide from sisal waste[J].Food Hydrocolloids,2014(39):10-18.
[11]DOU J,MENG Y H,LIU L,et al.Purification,characterization and antioxidant activities of polysaccharides from thinned-young apple[J].International Journal of Biological Macromolecules,2015,72:31-40.
[12]张惟杰.糖复合物生化研究技术[M].杭州:浙江大学出版社,1999:47.
[13]WANG X,L譈X.Characterization of pectic polysaccharides extracted from apple pomace by hot-compressed water[J].Carbohydrate Polymers,2014,102(102):174-184.
[14]DU Y Q,WANG J H.Structural analysis and antioxidant activities of polysaccharideisolated from Jinqian mushroom[J].International Journal of Biological Macromolecules,2014,64(2):63-68.
[15]JIA D,ZHANG J,LAN R,et al.A simple preparative method for isolation and purification of polysaccharides from mulberry(Morus alba L.)leaves[J].International Journal of Food Science&Technology,2013,48(6):1275-1281.
[16]YING Z,HAN X,LI J,et al.Ultrasound-assisted extraction of polysaccharides from mulberry leaves[J].Food Chemistry,2011,127(3):1273-1279.
[17]LI N S,YAN C Y,HUA D H,et al.Isolation,purification,and structural characterization of a novel polysaccharide from Ganoderma capense[J].International Journal of Biological Macromolecules,2013,57(6):285-290.
[18]SHARMA R,AHUJA M.Thiolated pectin:synthesis,characterization andevaluation as a mucoadhesive polymer[J].Carbohydrate Polymers,2011,85(3):658-663.
[19]张淑娟,徐怀德,米林峰.光皮木果汁体外抗氧化活性研究[J].食品科学,2011,32(21):56-61.
[20]GRIGORAS C G,DESTANDAU E,FOUG?RE L,et al.Evaluation of apple pomace extracts as a source of bioactive compounds[J].Industrial Crops and Products,2013,49(4):794-804.
[21]盛文静,薛长湖,赵庆喜,等.不同海参多糖的化学组成分析比较[J].中国海洋药物杂志,2007,26(1):44-51.
[22]ZHU Z Y,NIAN L,SI C L,et al.Structure and antitumor activity of a high-molecular-weight polysaccharide from cultured mycelium of Cordyceps gunnii[J].Carbohydrate Polymers,2012,88(3):1072-1076.
[23]加藤宏治,新田正已,原田政子,等.糖质的红外线吸收光谱[J].静冈大学农学部研究报告,1972,22:77-85.
[24]张颖,曾艳,张丽姣,等.蛹虫草菌糠多糖的分离纯化及结构组成分析[J].食品科学,2014,35(13):54-59.
[25]SHEN S,CHENG H R,LI XU,et al.Effects of extraction methods on antioxidant activities of polysaccharides from camellia seed cake[J].European Food Research and Technology,2014,238(6):1015-1021.
[26]LIU L,CAO J,HUANG J,et al.Extraction of pectins with different degrees of esterification from mulberry branch bark[J].Bioresource Technology,2010,101(9):3268-3273.
[27]IIJIMA M,NAKAMURA K,HATAKEYAMA T,et al.Phase transition of pectin with sorbed water[J].Carbohydrate Polymers,2000,41(1):101-106.
[28]KANG,DUCK-HEE R N.Oxidative stress,DNA damage,and breast cancer[J].AACN Advanced Critical Care,2002,13(4):540-549.
[29]G譈L覶IN I.Antioxidant and antiradical activities of L-carnitine[J].Life science,2006,78(8):803-811.
[30]CHEN G T,MA X M,LIU S G,et al.Isolation,purification and antioxidant activities of polysaccharides from Grifola frondosa[J].Carbohydrate Polymers,2012,89(1):61-66.
[31]WANG Y F,YANG Z W,WEI X L,et al.Antioxidant activities potential of tea polysaccharide fractions obtained by ultra filtration[J].International Journal of Biological Macromolecules,2012,50(3):558-564.
[32]杨江涛,杨娟,谢红,等.刺梨多糖粗品与纯品体外抗氧化作用[J].食品工业科技,2008,13(2):94-96.