红毛藻多糖的提取纯化及其性质的研究
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摘要
红毛藻(Bangia fusco-purpurea)是我国福建南日岛一种营养价值高且多糖含量丰富的经济养殖海藻。本文系统地研究了红毛藻多糖的提取、分离纯化工艺及其理化性质,并对红毛藻多糖的体外抗氧化活性做了初步的探索。
以多糖的提取率与硫酸根含量为指标,比较了热水、冷水、稀酸、稀碱提取红毛藻多糖的效果,发现用热水提取红毛藻多糖提取率与硫酸根含量最高,故选用热水作为提取溶剂。通过单因素与正交实验确定了热水提取的最佳工艺参数为:浸提温度90℃,浸提时间2小时,料液比1:30,浸提次数3次,在此优化的条件下得到多糖提取率为5.97%;采用超声波辅助热水提取红毛藻多糖,通过单因素与正交实验确定了最佳工艺参数为浸提温度为80℃,浸提时间为80 min,料液比1:70,超声波功率200 W,在最佳条件下多糖的提取率为9.69%;用乙醇沉淀多糖时乙醇的最佳终浓度为75%,多糖提取液的最佳浓缩比为1:3,此时的沉淀率为90.77%。
用热水提取的红毛藻粗多糖中含有大量的杂蛋白,本文比较了Sevag法、TCA法、中性蛋白酶法及碱性蛋白酶法去除红毛藻粗多糖中的蛋白质,使用这四种脱蛋白方法的脱除率分别为78.50%、56.56%、90.67%和96.77%,多糖的损失率分别为56.50%、10.63%、7.95%、9.80%,可见,酶法脱蛋白的效果好且多糖的损失较少。
用阴离子交换柱DEAE Cellulose-52对红毛藻多糖进行分级分离,分别用0 mol/L、0.1 mol/L、0.3 mol/L、0.5 mol/L及2 mol/L的NaCl进行分段梯度洗脱,在盐离子浓度为0.1 mol/L、0.3 mol/L和0.5 mol/L处分别得到三个多糖峰并命名为BPⅠ、BPⅡ和BPⅢ。接着用凝胶过滤柱Sephadex G-100对上述三个组分的多糖继续进行纯化,结果表明,凝胶过滤柱Sephadex G-100可对BPⅠ、BPⅡ和BPⅢ三个多糖组分中的小蛋白有效去除,但BPⅠ中还有一部分蛋白与多糖结合在一起,可能是一种糖蛋白。经紫外光谱分析与冻融分析均表明上述三种多糖为较纯的多糖组分,把Sephadex G-100柱纯化的多糖组分分别命名为BPSⅠ、BPSⅡ和BPSⅢ。
测定三种多糖的硫酸根含量,发现BPSⅢ的硫酸根含量最高,其次是BPSⅡ,而BPSⅠ含量最低,且多糖的纯度越高硫酸根含量越高。气相色谱分析结果表明,BPSⅠ、BPSⅡ的单糖组分主要由半乳糖组成,而BPSⅢ除含有半乳糖外还含有少量木糖,其摩尔比为11: 1。分析三种多糖的红外光谱可知三种多糖均含有β-D-半乳糖,硫酸根的位置均在C6位上,BPSⅠ中含有3,6-内醚,BPSⅡ和BPSⅢ不含3,6-内醚。
测定红毛藻粗多糖(BP)、BPSⅠ、BPSⅡ和BPSⅢ四种多糖的体外抗氧化活性,结果表明,BP、BPSⅠ、BPSⅡ、BPSⅢ对·OH自由基的半数抑制浓度(IC50)分别为4.98 mg/mL、1.22 mg/mL、13.86 mg/mL和1.78 mg/mL;对DPPH自由基的IC50值分别为0.70 mg/mL、1.05 mg/mL、0.58 mg/mL和0.55 mg/mL;对脂质过氧化体的IC50值分别为5.37 mg/mL、2.83 mg/mL、24.02 mg/mL以及1.60 mg/mL,而对O-2的抑制作用却并不显著。
Bangia fusco-purpurea is a kind of red alga which is an economic aquaculture alga in Fujian province. In this paper, the extraction process, purification method , the physical and chemical characters of Bangia fusco-purpurea polysaccharides (BP) were systematically studied, and its antioxidant activity in vitro were also been preliminarily explored.
We compared different solvents effect on extraction yield and sulfate content of polysaccharides, the method of hot water, cold water, dilute acid and dilute alkali were used in the experiment. The result showed that hot water is the best solvent. According to single factor and orthogonal test, the optimum conditions of extract process were as follows: extracting temperature was 90℃, extracting time was 2 h, the ratio of alga to solvent was 1:30 and extracting times was 3. The maximum yield of polysaccharide was 5.97% under this optimized conditions; Ultrasonic method was used as an auxiliary method in the extraction of polysaccharide. According to single facter and orthogonal test, the optimum conditions of extractions were: extracting temperature was 80℃, extracting time was 80 minutes, the ratio of alga to solvent was 1:70 and the power of ultyasonic was 200 W. The maximum yield of polysaccharide was 9.69% under this condition; The ethanol concentration of 75% and the concentrate ratio of extracting solution of 1: 3 were optimized for polysaccharide precipitation. Polysaccharide extracted by hot water contains a large amount of protein. Sevag method, TCA method, neutral protease method and alkaline protease method were used to remove the protein. The deproerinization efficiency was 78.50%、56.56%、90.67% and 96.77% respectively, and the loss of polysaccharide was 56.50%、10.63%、7.95% and 9.80% respectively. It is clear to see that the application of protease method in deproteinizaiton was much more efficiency and had less loss of polysaccharide.
BP was separated by DEAE Cellulose-52 column, The elution was performed by step-wise process using the same buffer containing different concentrations of NaCl: 0.1 mol/L, 0.3 mol/L , 0.5 mol/L and 2 mol/L, respectively. and three fractions were obtained under the concentrations of 0.1 mol/L, 0.3 mol/L, 0.5 mol/L, which were named BPⅠ、BPⅡand BPⅢ. The Sephadex G-100 column was applied for further purification. The result showed that Sephadex G-100 could remove the protein from BPⅠ、BPⅡand BPⅢeffectively , but there was some protein combine with BPⅠ, it may be a kind of glycoprotein. Ultraviolet spectrum analysis and freeze-thaw analysis showed that these fractions were purified polysaccharide, and named as BPSⅠ、BPSⅡand BPSⅢ.
The content of sulfate was different amount different fractions, BPSⅢhad highest level of sulfate, the next was BPSⅡa nd the BPSⅠhad the lowest level of sulfate. The content of sulfate was also positive correlation with its purity. Gas chromatography analysis showed that the mainly monomers of BPSⅠa nd BPSⅡwas galactose, the BPSⅢalso contained some xylose except galatose. According to IR spectra, it could be confirmed that the ploysacchardes part of BPSⅠwas composed by 6-sulfate-β-D-galactose and 3, 6-anhydro-glaetose, BPSⅡa nd BPSⅢwere composed by 6-sulfurde–D-galactose, which did not contain 3, 6-anhydro-glaetose.
Antioxidant activity in vitro of BP、BPSⅠ、BPSⅡand BPSⅢwas determined, the result showed that the IC50 on·OH was 4.98 mg/mL、1.22 mg/mL、13.86 mg/mL and 1.78 mg/mL respectively, the IC50 on DPPH was 0.70 mg/mL、1.05 mg/mL、0.58 mg/mL and 0.55 mg/mL respectively, the IC50 on liposome oxidation was 5.37 mg/mL、2.83 mg/mL、24.02 mg/mL and 1.60 mg/mL respectively, but there was no significant effect on the inhibition of O-2.
以多糖的提取率与硫酸根含量为指标,比较了热水、冷水、稀酸、稀碱提取红毛藻多糖的效果,发现用热水提取红毛藻多糖提取率与硫酸根含量最高,故选用热水作为提取溶剂。通过单因素与正交实验确定了热水提取的最佳工艺参数为:浸提温度90℃,浸提时间2小时,料液比1:30,浸提次数3次,在此优化的条件下得到多糖提取率为5.97%;采用超声波辅助热水提取红毛藻多糖,通过单因素与正交实验确定了最佳工艺参数为浸提温度为80℃,浸提时间为80 min,料液比1:70,超声波功率200 W,在最佳条件下多糖的提取率为9.69%;用乙醇沉淀多糖时乙醇的最佳终浓度为75%,多糖提取液的最佳浓缩比为1:3,此时的沉淀率为90.77%。
用热水提取的红毛藻粗多糖中含有大量的杂蛋白,本文比较了Sevag法、TCA法、中性蛋白酶法及碱性蛋白酶法去除红毛藻粗多糖中的蛋白质,使用这四种脱蛋白方法的脱除率分别为78.50%、56.56%、90.67%和96.77%,多糖的损失率分别为56.50%、10.63%、7.95%、9.80%,可见,酶法脱蛋白的效果好且多糖的损失较少。
用阴离子交换柱DEAE Cellulose-52对红毛藻多糖进行分级分离,分别用0 mol/L、0.1 mol/L、0.3 mol/L、0.5 mol/L及2 mol/L的NaCl进行分段梯度洗脱,在盐离子浓度为0.1 mol/L、0.3 mol/L和0.5 mol/L处分别得到三个多糖峰并命名为BPⅠ、BPⅡ和BPⅢ。接着用凝胶过滤柱Sephadex G-100对上述三个组分的多糖继续进行纯化,结果表明,凝胶过滤柱Sephadex G-100可对BPⅠ、BPⅡ和BPⅢ三个多糖组分中的小蛋白有效去除,但BPⅠ中还有一部分蛋白与多糖结合在一起,可能是一种糖蛋白。经紫外光谱分析与冻融分析均表明上述三种多糖为较纯的多糖组分,把Sephadex G-100柱纯化的多糖组分分别命名为BPSⅠ、BPSⅡ和BPSⅢ。
测定三种多糖的硫酸根含量,发现BPSⅢ的硫酸根含量最高,其次是BPSⅡ,而BPSⅠ含量最低,且多糖的纯度越高硫酸根含量越高。气相色谱分析结果表明,BPSⅠ、BPSⅡ的单糖组分主要由半乳糖组成,而BPSⅢ除含有半乳糖外还含有少量木糖,其摩尔比为11: 1。分析三种多糖的红外光谱可知三种多糖均含有β-D-半乳糖,硫酸根的位置均在C6位上,BPSⅠ中含有3,6-内醚,BPSⅡ和BPSⅢ不含3,6-内醚。
测定红毛藻粗多糖(BP)、BPSⅠ、BPSⅡ和BPSⅢ四种多糖的体外抗氧化活性,结果表明,BP、BPSⅠ、BPSⅡ、BPSⅢ对·OH自由基的半数抑制浓度(IC50)分别为4.98 mg/mL、1.22 mg/mL、13.86 mg/mL和1.78 mg/mL;对DPPH自由基的IC50值分别为0.70 mg/mL、1.05 mg/mL、0.58 mg/mL和0.55 mg/mL;对脂质过氧化体的IC50值分别为5.37 mg/mL、2.83 mg/mL、24.02 mg/mL以及1.60 mg/mL,而对O-2的抑制作用却并不显著。
Bangia fusco-purpurea is a kind of red alga which is an economic aquaculture alga in Fujian province. In this paper, the extraction process, purification method , the physical and chemical characters of Bangia fusco-purpurea polysaccharides (BP) were systematically studied, and its antioxidant activity in vitro were also been preliminarily explored.
We compared different solvents effect on extraction yield and sulfate content of polysaccharides, the method of hot water, cold water, dilute acid and dilute alkali were used in the experiment. The result showed that hot water is the best solvent. According to single factor and orthogonal test, the optimum conditions of extract process were as follows: extracting temperature was 90℃, extracting time was 2 h, the ratio of alga to solvent was 1:30 and extracting times was 3. The maximum yield of polysaccharide was 5.97% under this optimized conditions; Ultrasonic method was used as an auxiliary method in the extraction of polysaccharide. According to single facter and orthogonal test, the optimum conditions of extractions were: extracting temperature was 80℃, extracting time was 80 minutes, the ratio of alga to solvent was 1:70 and the power of ultyasonic was 200 W. The maximum yield of polysaccharide was 9.69% under this condition; The ethanol concentration of 75% and the concentrate ratio of extracting solution of 1: 3 were optimized for polysaccharide precipitation. Polysaccharide extracted by hot water contains a large amount of protein. Sevag method, TCA method, neutral protease method and alkaline protease method were used to remove the protein. The deproerinization efficiency was 78.50%、56.56%、90.67% and 96.77% respectively, and the loss of polysaccharide was 56.50%、10.63%、7.95% and 9.80% respectively. It is clear to see that the application of protease method in deproteinizaiton was much more efficiency and had less loss of polysaccharide.
BP was separated by DEAE Cellulose-52 column, The elution was performed by step-wise process using the same buffer containing different concentrations of NaCl: 0.1 mol/L, 0.3 mol/L , 0.5 mol/L and 2 mol/L, respectively. and three fractions were obtained under the concentrations of 0.1 mol/L, 0.3 mol/L, 0.5 mol/L, which were named BPⅠ、BPⅡand BPⅢ. The Sephadex G-100 column was applied for further purification. The result showed that Sephadex G-100 could remove the protein from BPⅠ、BPⅡand BPⅢeffectively , but there was some protein combine with BPⅠ, it may be a kind of glycoprotein. Ultraviolet spectrum analysis and freeze-thaw analysis showed that these fractions were purified polysaccharide, and named as BPSⅠ、BPSⅡand BPSⅢ.
The content of sulfate was different amount different fractions, BPSⅢhad highest level of sulfate, the next was BPSⅡa nd the BPSⅠhad the lowest level of sulfate. The content of sulfate was also positive correlation with its purity. Gas chromatography analysis showed that the mainly monomers of BPSⅠa nd BPSⅡwas galactose, the BPSⅢalso contained some xylose except galatose. According to IR spectra, it could be confirmed that the ploysacchardes part of BPSⅠwas composed by 6-sulfate-β-D-galactose and 3, 6-anhydro-glaetose, BPSⅡa nd BPSⅢwere composed by 6-sulfurde–D-galactose, which did not contain 3, 6-anhydro-glaetose.
Antioxidant activity in vitro of BP、BPSⅠ、BPSⅡand BPSⅢwas determined, the result showed that the IC50 on·OH was 4.98 mg/mL、1.22 mg/mL、13.86 mg/mL and 1.78 mg/mL respectively, the IC50 on DPPH was 0.70 mg/mL、1.05 mg/mL、0.58 mg/mL and 0.55 mg/mL respectively, the IC50 on liposome oxidation was 5.37 mg/mL、2.83 mg/mL、24.02 mg/mL and 1.60 mg/mL respectively, but there was no significant effect on the inhibition of O-2.
引文
[1]吕惠敏,张侃.海藻的利用与开发[J].食品科学,1998,(6):29-30.
[2]孟春,郭养浩,石贤爱,等.海藻多糖生物活性及分子修饰.中国生物工程杂志,2004,24(3):35-39.
[3] PATTERSON G M L, LARSEN L K, MOORE R E. Bioactive natural products from blue-green algae[J]. Journal of Applied Phycology, 1994, (6):151-157.
[4] SHIDDHANTA A K. Marine algal polysaccharides functions and utilization[M]. India: Surya International Publications, 1995:125-131.
[5] KAY R A. Microalgae as food and supplement [J]. Criti Rev Food Sci Nutr, 1991, 30(6): 555-573.
[6]纪明侯.海藻化学[M].北京:科学出版社,1997.
[7]张惟杰.糖复合物生化研究技术[M].第二版.杭州:浙江大学出版社,2003:30-120.
[8]王长云,管华诗.多糖抗病毒作用研究进展II硫酸多糖抗病毒作用[J].生物工程进展,2000,20(2):3-8.
[9]吴永沛,邱晓燕,张东,等.海带提取岩藻聚糖的研究[J].食品科学,2003,24(10):78-80.
[10] HEO S J, LEE K W, JEON Y J. Antioxidant activity of enzymatic extracts from browm seaweeds [J].Algae, 2005, 96(14):1616-1623.
[11]卢睿春,刘婉乔,侯振建,等.酶法提取马尾藻硫酸多糖的研究[J].中国海洋药物,1997,(2):10-13.
[12]刘轲,王琪琳,吕辉,等.海带硫酸多糖的提取、纯化及其理化分析[J].中国生化药物杂志,2002,23(3):114-116.
[13]刘慧燕,赵谋明.江蓠低分子量多糖的提取以及抗氧化活性研究[J].食品工业科技,2005,26(3):67-69.
[14]赵兵,王玉春,孙学兵,等.循环气升式超声破碎鼠尾藻提取海藻多糖[J].中国海洋药物,1999,(4):19-23.
[15]杨文杰,黄惠华,李琳,等.螺旋藻多糖的水提微波辅助提取的比较[J].食品工业科技,2003,24(8):40-42.
[16]徐惠,于志清,苏富强,等.螺旋藻牯多糖的分离及其免疫学作用[J].中国生化药物杂志,1997,18(2):72-75.
[17]高向东,盛海林,潘秋文,等.均匀设计和球面对称设计在螺旋藻多糖最佳工艺研究中的应用比较[J].中国药科大学学报,1997,28(3):171-173.
[18] TABBA H D, CHANG R S, SMITH K M.Isolation, purification and partial characterization of prunellin, an anti-HIV component from aqueous extract of prunella vulgaris[J].Antiviral Res, 1989, 11(5-6): 283.
[19]强亦忠,王崇道,邵源,等.海藻硫酸多糖的制备及其性质研究[J].苏州大学学报,2003,23(4):391-393.
[20]章银良,李红旗,高峻,等.螺旋藻多糖提取新工艺的研究[J].食品与发酵工业,1999,259(2):15-18.
[21] COLLIDE S, BOISSO-VIDAL C, JOZEFONVICA J. A low molecufar weight fucoldan fraction from the brown seaweed Pelvetia canaliculata[J]. Phytochemistry, 1994, 35(3): 697.
[22]过菲,许时婴,林之川.超滤技术在羊栖菜粗多糖提取工艺中的应用[J].工艺技术,2002,23(10):50-51.
[23] BIMALENDU RAY. Polysaccharides from Enteromorpha compressa: Isolation, purification and structural features[J]. Carbohydrate Polymers, 2006, 66(3): 408-416.
[24]伍志春,欧阳藩,房燕丽,等.非水溶性有机试剂萃取海藻多糖[J].高技术通讯,2000,(8):19-21.
[25] CHEVOLOT, FOUCAULT A, CHAUBERT F. Further data on the structure of brown seaweed fucans: relationships with anticoagulant activity [J]. Carbohydrate Research, 1999, 319(1-4):154-165.
[26] GIROND S, CRANCE J M, VAN CUYCK-GANDRE H, et al. Antiviral activity of carrageenan on hepatitis A virus replication in cell cluture [J]. Res Virol, 1991, 142(4): 261-270.
[27] HAMASUNA R, EIZURU Y, MINAMISHIMA Y. Inhibition by iota-carrgeenan of the spread of murine cytomegalovirus from the peritoneal cavity to the blood plasma [J]. J Gen Virol, 1994, 75(1):111-116.
[28] HASUI M, MATSUDA M, OKUTANI K, et al. In vitro antiviral activities of sulfated polysaccharices from a marine microalga(Cochlodinium polykrikoides) against human immunodeficiency virus and other enveloped viruses [J]. International Journal of Biological Macromolecules, 1995, 17(5): 293-297.
[29] CARLUCCI M J, CIANCIA M, MATULEWICZ M C, et al. Antiherpetic activity and mode of action of matural carrageenans of diverse structural types [J]. Antiviral Research, 1999, 43(2):93-102.
[30] DAMONTE E, NEYTS J, PUJOL C A, et al. Antiviral activity of a sulphated polysaccharide from the red seaweed Nothogenia fastigiata [J]. Biochemical Pharmacology, 1994, 47(12): 2187-2192.
[31] PUJOL C A, COTO C E, DAMONTE E B.Determination of the antiviral activity of a naturally occurring sulftated xylomannan under various experimental conditions [J]. Rev Agent Microbiol, 1995, 27(2):91-98.
[32] CRANCE J M, GRATIER D, GUIMT J, et al. Inhibition of sacdfly fever Sicilian virus (phlebovirus) replication in vitro by antiviral compounds [J]. Res Virot, 1997, 148(5):353-365.
[33]王长云,管华诗.多糖抗病毒作用研究进展Ⅲ.卡拉胶及其抗病毒作用[J].生物工程进展,2000a,20(3):39-42.
[34] SUTAPA M, PRODYUT K, GHOSAL G A, et al. Isolation, chemical investigation and antiviral activity of polysaccharides from Gracilaria corticata (Gracilariaceae, Rhodophyta) [J]. International Journal of Biological Macromolecules, 2002, 31(1-3):87-95.
[35] HOSHINO T, HAYASHI T, HAYASHI K, et al. An antivirally active sulfated polysaccharide from Sargassum horneri(TURNER) C. AGARDH [J]. Biological & pharmaceutical bulletin, 1998, 21(7):730-734.
[36] PREEPRAME S, HAYASHI K, LEE JB, et al. A novel antiviraliy active fucan sulfate derived from an edible brown alga, Sargassum horneri [J]. Chemical & pharmaceutical bulletin (Tokyo), 2001, 49(4):484-485.
[37] REN D L, WANG J Z, AMANO H, et al. The effects of an algal polysaccharide from Gloiopeltis tenax on transplantable tumors and immune activities in mice [J]. Phanta Med, 1995, 61(2):120-124.
[38] COOBE D. Antitumor activity of ediblemmarine algae: effect of crude fucoid an fraction prepared from edible seaweeds against tumor [J]. International Journal of Cancer, 1987, (39):82-85.
[39] ITOCH H, NODA H, AMANO H, et al. Immunological analysis of inhibition of lung metastases by fucoidan (GIV-A) prepared from brown seaweed Sargassum thunbergii [J]. Anticancer Research, 1995, 15(5B): 1937-1947.
[40] ZHUANG C, ITOH H, MIZUNO T, et al. Antitumor active fucoidan from the brown seaweed, umitoranoo(Sargassum thunbergii) [J]. Biosci Biotechnol Biochem, 1995, 59(4):563-567.
[41]西泽一俊.海藻生物活性物质.食品开发(日),1980,18(11):28-30.
[42] OKAI Y, ISHIZAKA S, HIGASHI-OKAI K. Detection of immunomodulating activities in an extract of Japanese edible seaweed, Laminaria Japonica(Makonbu) [J]. Journal of the Science of Food and Agriculture, 1996, 72(4):455-460.
[43]董为人,田雪梅.补体活化在动脉粥样硬化形成中的意义[J].国外医学免疫分册,1999,22(1):47-50.
[44] KUZNETSOVA T A, ZAPOROZHETS T S, BESEDNOVA N N, et al. Immunostimulating and anticoagulating activity of fucoidan from browm algae Fucus evanescens of Okhotskoe sea [J]. Antbiot Khimioter, 2003, 48(4):11-13.
[45]张全斌,李大新.坛紫菜多糖对脾细胞活性的影响[J].中国海洋药物,2003,96(6):14-18.
[46]王常青,杨桂兰.两种海藻多糖对大鼠脂质代谢和血小板功能的比较[J].中华预防医学杂志,1997,31(6):342-345.
[47]王艳梅,李智恩,牛锡珍,等.孔石莼多糖降血脂活性的初步研究[J].中国海洋药物,2003,22(2):33-35.
[48]左绍远,钱金,万顺康,等.螺旋藻多糖降血糖活性实验研究[J].时珍国医国药,2000,11(8):677-678.
[49]王兵,李靖,马舒冰.羊栖菜多糖降血糖的实验研究[J].中国海洋药物,2000,(3):33-35.
[50]李八方,毛文君,曹立民.羊栖菜多糖对高血脂模型大鼠血脂的影响[J].中国水产科学,2000,7(2):56-58.
[51] CHURH F C, MEADE J B, TREANOR R E, et al. Antithrombin activity of fucoidan.The interaction of fucoidan with heparin cofactorⅡ. AntithrombinⅢ, and thrombin [J].The Journal of Biological Chemistry, 1989,264(6):3618-3623.
[52] TAKASHI N, AKIHIRO F, TERUKAZU N, et al. Inhibition of the generation of thrombin and factor Xaby a fucoindan from the brown seaweed Ecklonia kurome [J]. Thrombosis Research, 1999, 96(1): 37-49.
[53] GRAUFFEL, KLOAREG, MABEAU, et al. New natural polysaccharides with potent antithrombic activity: fucans from brown algae [J].Biomaterials, 1989, 10(6): 363-368.
[54] COLLIEC, FISCHER, TAPON-BRETAUDIERE, et al. Anticoagulant properties of a fucoidan fraction [J]. Thrombosis Research. 1991, 64(2): 143-154.
[55] MATSUBARA, BACIC. Anticoagulant properties of a sulfated galactan preparation from a marine green alga Codium cylindricum [J]. International Journal of Biological Mzcromolecules, 2001, 28(5): 395-399.
[56]王春玲,张全斌.褐藻多糖硫酸酯抗凝血活性的研究[J].中国海洋药物,2005,24(5):36-38.
[57] MARIANA S. P, ANA-CRISTINA E. S. VILELA-SILVA, ANA-PAULA VALENTE, et al. A 2-sulfated, 3-linked-α-Lgalactan is an anticoagulant polysaccharide [J]. Carbohydrate Research. 2002, 337(21-23), 2231–2238.
[58]王静凤,张学成,姜国良,等.枝管藻多糖的提取及其抗凝血活性的初步研究[J].青岛海洋大学学报,2003,33(1):75-79.
[59] Hayakawa Y , Hayashi T, Lee J, et al. Inhibition of thrombin by sulfated polysaccharides isolated from green algae[J]. Biochimaica et Biophysica Acta, 2000, 1543(1):86-94.
[60] Yasantha A, Kil-Nam K, You-Jin J. Antiproliferative and antioxidant properties of an enzymatic hydrolysate from browm alga, Ecklonia cava [J]. Food and Chemical Toxicology, 2006, 44(7):1065-1074.
[61] Hu, J. F., Geng, M.Y., Zhang, J. T., Jiang, H. D.. An in vitro study of the structure-activity relations of sulfated polysaccharide from brown algae to its antioxidant effect [J]. Journal of Asian Natural Product Reports, 2001, 3(4): 353-358.
[62] Zhang Q., Li, N., Zhou, G., et al. In vivo antioxidant activity of polysaccharide fraction from Porphyra haitanesis (Rhodephyta) in aging mice [J]. Pharmacological Research, 2003,48(2): 151-155.
[63] Zhang Q, Yu P, Li Z, et al,. Antioxidant activities of sulfated polysaccharide fractions from Porphyra haitanesis[J]. Journal of Applied Phycology , 2003,15(4):305-310.
[64]项华,赵健忠,柏林.海藻多糖抗突变性研究[J].癌变·畸变·突变,2001,13(1):42-44.
[65]陈美珍,余杰,龙梓洁,等.龙须菜多糖抗突变和清除自由基作用的研究[J].食品科学,2005,26(7):219-222.
[66]余杰,许肇成,颜璐璐,等.海萝多糖抗突变与抗肿瘤作用的研究[J].汕头大学学报,2007,22(2):59-63.
[67]庞辉,汤桂芳,玉艳红.螺旋藻多糖的抗溃疡作用及机制[J].广西医学,2006,28(11):1772-1773.
[68] Maisin JR. Bacq and Alecander Award lecture-chemical radioprotection: past, resent, and future prospects[J]. Int J Radiat Biol, 1998, 73(4): 443.
[69]吴晓旻,杨明亮,黄晓兰.海带多糖的抗辐射作用与脾细胞调亡[J].武汉大学学报,2004,25(3):239-241.
[70]罗琼,吴晓旻,杨明亮,等.海带多糖的抗辐射作用与淋巴细胞凋亡关系研究[J].营养学报,2004,26(12):471-473.
[71] Crist ,R. H., K. Oberholser, N. Shank, et al. Nature of Bonding Between Metallic Ions and Algal Cell Walls [J]. Environ. Science & Technol, 1981, 15(10):1212-1217.
[72] Tarlan E, Yetis U, Dilek F B. Algal treatment of pulp and paper industry wastewasters in SBR systems[J]. Water Science and Technology, 2002, 45(12):151-158.
[73] SAEKI Y, KATO T, OKUDA K. Inhibitory effects of funoran on the adherence and colonization of oral bacteria[J]. Bullentin of Tokyo Dental College, 1996, 37(2):77-92.
[74] MATSUI M S, MUIZZUDDIN N, ARAD S, et al. Sulfated polysaccharides from red microalgae have anti-inflammatory properties in vitro and in vivo [J]. Appl Biochem Biotechnol, 2003, 104(1):13-32.
[75] HINCHA D K, ZUTHER E, HELLWEGE E M, et al. Specific effects fructo-and glucooligosaccharides in the preservation of liposomes during drying [J]. Glycobiology, 2002, 12(2):103-110.
[76] EHRESMANN D W, DEIG E F, HATCH M T, et al. Dextran sulfate, a potent anti-HIV agent in vitro having synergism with sidovudine [J]. Lancet, 1987, 13(1):1379-1386.
[77] NARDELLA A, CHAUBET F, BOISSON-VIDAL, et al. Anticoagulant low molecular weight fucans produced by radical process and ion exchange chromatography of high molecular weight fucans extracted from the brown seaweed Ascophyllum nodosum [J]. Carbohydrate Research, 1996, 289(8): 201-208.
[78] YASANTHA A, WON-KYO J, THAVA V, et al. An anticoagulative polysaccharide from an enzymatic hydrolysate of Ecklonia cava [J]. Carbohydrate Polymers ,2006, 66(2): 184-191.
[79] SHANMUGAM S, MODY K H. Heparonid active sulfated polysaccharides from marine algae as potential blood coagulant agents [J]. Currant Sciences, 2000, (79): 1672-1683.
[80] SUSANNE A, GERHARD F. Characterization of the Anticoagulant Actions of a Semisynthetic Curdlan Sulfate[J]. Thrombosis Research, 2000, 99 (4): 377-388.
[81]刘志峰,宫晓黎,魏淑贞.五种海藻多糖体外抗血小板聚集作用观察[J].中国海洋药物,2001,20(2):36-38.
[82] TSIPALI E, WHALEY S, KALBFLEISCH J, et al. Glucans exhibit weak antioxidant activity, but stimulate macrophage free radical activity [J]. Free Radical Bicology and Medicine, 2001, 30(4):393-402.
[83] GROTH T, WAGENKNECH T. Anticoagulant potential of regioseselective Derivatized cellulose [J]. Biomaterials, 2001, 22(8): 2719-2729.
[84] MARIA G P, NORMA M B B, MARCI R S, et al. Structure and anticoagulant activity of a sulfated galactan from the red alga, Gelidium crinale. Is there a specific structural requirement for the anticoagulant action? [J]. Carbohydrate Research, 2005, 340(12): 2015-2023.
[85] MULLOY B, MOURAO PAS, GRAY E, et al. Structure/function studies of anticoagulant sulphated polysaccharides using NMR [J]. Journal of biotechnology, 2000, 77(1): 123-135.
[86]诸葛健,赵振锋,方慧英.功能性多糖的构效关系[J].无锡轻工大学学报,2002,21(2):209-212.
[87]中国农业百科全书,水产业卷[M].北京:中国农业出版社,1994,218-219,748-750.
[88]曾呈奎.中国经济海藻志[M].北京:科学出版社,1962:158-212.
[89]黄文凤,黄建明,董飞强.红毛菜的营养成分特征和价值[J].海洋水产研究,1998,19(2):57-61.
[90]马家海,李水军,纪焕红,等.红毛菜的氨基酸和脂肪酸分析[J].中国海洋药物杂志,2002,5:40-42.
[91]食品中蛋白质的测定方法.中华人民共和国国家标准,GB/T 14771-1993.
[92]黄伟坤.食品检测与分析[M].北京:轻工业出版社.1997:49-51.
[93]夏敏.必需微量元素与人体健康[J].中国食物与营养,2003,(10):53-56.
[94]藻类制品卫生标准,GB-19643-2005.
[95]王进卿.膳食纤维的功能与应用[J].中国酿造,2003,(5):37-38.
[96] DODGSON K S, PRICE R G. A note on the determination of the ester sulfate content of sulfated polysaccharides[J]. Biochemistry Journal, 1962, 84(1): 106-110.
[97]李宏燕,樊君.大枣多糖的水提醇沉工艺研究[J].宁夏工程枝术,2005,4(3):265-267.
[98]刑小黑,吴明忠,朱述钧,等.灵芝多糖化学研究[J].中国食用菌,1996,15(3):1-16.
[99]刘晓红,肖凯军.金樱子粗多糖的脱蛋白研究[J].食品科技,2007,(1):192-194.
[100]周存山,马海乐,胡文彬.条斑紫菜多糖提取工艺的优化[J].农业工程学报,2006,22(9):194-197.
[101]余华.海带多糖中蛋白质去除方法的对比研究[J].成都大学学报,2005,24(4):265-268.
[102]李瑞军,李德耀,张现峰,等.大孔吸附树脂法去除淫羊藿多糖中蛋白的研究[J].高等学校化学学报,2006,27(1):60-70.
[103]贾建波,李相前,杨文.桑黄多糖分离纯化及其结构初步鉴定[J].食品科学,2006,27(12):446-450.
[104]黄纯,马驰,宋慧智.亮菌多糖提取中脱蛋白和脱色方法比较[J].药学与临床研究,2007,15(1):45-46.
[105]陈小萍,张卫明,史劲松,等.茶树花水提多糖的精制工艺初探[J].食品科技,2007,32(4):72-75.
[106]何传波,陈玲,李琳.巴戟天多糖脱蛋白方法的研究[J].食品科技,2005(6):25-27.
[107]朱美静,童群义.猴头多糖脱蛋白方法的研究[J].河南工业大学学报,2005,26(4):25-27.
[108]盛家荣,曾令辉,翟春,等.多糖的提取、分离及结构分析[J].广西师院学报(自然科学版),1999,16(4):49-54.
[109]陈京,熊耀康.多糖类药物的质量标准研究现状及其进展[J].浙江中医学院学报,2004,28(3):77-80.
[110]张丽萍,李森,黄丽萍.金顶侧耳酸提水溶性多糖的研究——PC-3的分离、纯化与结构确定[J].菌物学报,1993,12(2):158-162.
[111]康琰琰,王一飞,朱良,等.裙带菜茎中硫酸多糖的分离纯化及分析鉴定[J].中药材,2005,28(9):769-771.
[112]黄益丽,郑天凌.海洋生物活性多糖的研究现状与展望[J].海洋科学,2004,28(4):58-61.
[113]张承圭,王传怀,袁玉荪,等.生物化学仪器分析及技术[M].北京:高等教育出版社,1990:84-90.
[114]刘成梅,付桂明,游海,等.百合多糖的纯化与化学结构鉴定研究[J].食品科学,2002,23(5):114-117.
[115]马丽,覃小林,刘雄民,等.螺旋藻多糖的纯化和效液相色谱分析[J].化工技术与开发,2004,33(2):29-32.
[116]董洪新,刘新海,李焕艺.灵芝多糖生物活性物质的研究现状及进展[J].当代生态农业,2003,(1):28-30.
[117] XIN X L, LIU C H. Progress in the study of antioxidant effects of Chinese herbal polysaccharides [J]. Journal of Beijing University of Traditional Chinese Medicine, 2000, 23(5): 54-55.
[118] HALLIWELL B, GUTTEIDE J M C, ARUMOA O I. The deoxyribose method: A simple“Test-Tube”assay for determination of rate constants for reactions of hydroxyl radicals [J]. Analytical Biochemistry, 1987, 165(1): 215-219.
[119]吉林市卫生防疫站.GB/T 5009.171-2003保健食品中超氧化物歧化酶(SOD)活性的测定[S].中国国家标准化管理委员会.北京:中国标准出版社,2003:410-417.
[120] DUAN X J, ZHANG W W, LI X M, et al. Evaluation of antioxidant property of extract and fractions obtained from a red alga, Polysiphonia urceolata [J], Food Chemistry, 2006, 95(1): 37 .
[121]范晓,严小军.高分子量褐藻多酚抗氧化性质研究[J].水生生物学报,1999,23(5):494-499.
[122] YI O S, MEYER A S, FRANKED E N. Antioxidant Activity of Grape Extracts in a Lecithin Liposeme System[J]. JAOCS, 1997, 74(10): 130l-1306.
[123]晏文洁,李家璞.类黄酮抗氧化力与其结构之关系[J].台湾农业化学与食品科学,2000,38(1):80-88.
[124] CUZZOCREA S, RILEY D, CAPUTI A P, et al. Antioxidant therapy: a new pharmacological approach in shock, inflammation and ischemis/reperfusion injury[J]. Pharmacological reviews, 2001, 25(5): 135-139.
[125]钦传光,周军,赵文,等.泥鳅多糖清除活性氧和保护DNA链的作用[J].生物化学与生物物理学报,2001,33(2):215-218.
[2]孟春,郭养浩,石贤爱,等.海藻多糖生物活性及分子修饰.中国生物工程杂志,2004,24(3):35-39.
[3] PATTERSON G M L, LARSEN L K, MOORE R E. Bioactive natural products from blue-green algae[J]. Journal of Applied Phycology, 1994, (6):151-157.
[4] SHIDDHANTA A K. Marine algal polysaccharides functions and utilization[M]. India: Surya International Publications, 1995:125-131.
[5] KAY R A. Microalgae as food and supplement [J]. Criti Rev Food Sci Nutr, 1991, 30(6): 555-573.
[6]纪明侯.海藻化学[M].北京:科学出版社,1997.
[7]张惟杰.糖复合物生化研究技术[M].第二版.杭州:浙江大学出版社,2003:30-120.
[8]王长云,管华诗.多糖抗病毒作用研究进展II硫酸多糖抗病毒作用[J].生物工程进展,2000,20(2):3-8.
[9]吴永沛,邱晓燕,张东,等.海带提取岩藻聚糖的研究[J].食品科学,2003,24(10):78-80.
[10] HEO S J, LEE K W, JEON Y J. Antioxidant activity of enzymatic extracts from browm seaweeds [J].Algae, 2005, 96(14):1616-1623.
[11]卢睿春,刘婉乔,侯振建,等.酶法提取马尾藻硫酸多糖的研究[J].中国海洋药物,1997,(2):10-13.
[12]刘轲,王琪琳,吕辉,等.海带硫酸多糖的提取、纯化及其理化分析[J].中国生化药物杂志,2002,23(3):114-116.
[13]刘慧燕,赵谋明.江蓠低分子量多糖的提取以及抗氧化活性研究[J].食品工业科技,2005,26(3):67-69.
[14]赵兵,王玉春,孙学兵,等.循环气升式超声破碎鼠尾藻提取海藻多糖[J].中国海洋药物,1999,(4):19-23.
[15]杨文杰,黄惠华,李琳,等.螺旋藻多糖的水提微波辅助提取的比较[J].食品工业科技,2003,24(8):40-42.
[16]徐惠,于志清,苏富强,等.螺旋藻牯多糖的分离及其免疫学作用[J].中国生化药物杂志,1997,18(2):72-75.
[17]高向东,盛海林,潘秋文,等.均匀设计和球面对称设计在螺旋藻多糖最佳工艺研究中的应用比较[J].中国药科大学学报,1997,28(3):171-173.
[18] TABBA H D, CHANG R S, SMITH K M.Isolation, purification and partial characterization of prunellin, an anti-HIV component from aqueous extract of prunella vulgaris[J].Antiviral Res, 1989, 11(5-6): 283.
[19]强亦忠,王崇道,邵源,等.海藻硫酸多糖的制备及其性质研究[J].苏州大学学报,2003,23(4):391-393.
[20]章银良,李红旗,高峻,等.螺旋藻多糖提取新工艺的研究[J].食品与发酵工业,1999,259(2):15-18.
[21] COLLIDE S, BOISSO-VIDAL C, JOZEFONVICA J. A low molecufar weight fucoldan fraction from the brown seaweed Pelvetia canaliculata[J]. Phytochemistry, 1994, 35(3): 697.
[22]过菲,许时婴,林之川.超滤技术在羊栖菜粗多糖提取工艺中的应用[J].工艺技术,2002,23(10):50-51.
[23] BIMALENDU RAY. Polysaccharides from Enteromorpha compressa: Isolation, purification and structural features[J]. Carbohydrate Polymers, 2006, 66(3): 408-416.
[24]伍志春,欧阳藩,房燕丽,等.非水溶性有机试剂萃取海藻多糖[J].高技术通讯,2000,(8):19-21.
[25] CHEVOLOT, FOUCAULT A, CHAUBERT F. Further data on the structure of brown seaweed fucans: relationships with anticoagulant activity [J]. Carbohydrate Research, 1999, 319(1-4):154-165.
[26] GIROND S, CRANCE J M, VAN CUYCK-GANDRE H, et al. Antiviral activity of carrageenan on hepatitis A virus replication in cell cluture [J]. Res Virol, 1991, 142(4): 261-270.
[27] HAMASUNA R, EIZURU Y, MINAMISHIMA Y. Inhibition by iota-carrgeenan of the spread of murine cytomegalovirus from the peritoneal cavity to the blood plasma [J]. J Gen Virol, 1994, 75(1):111-116.
[28] HASUI M, MATSUDA M, OKUTANI K, et al. In vitro antiviral activities of sulfated polysaccharices from a marine microalga(Cochlodinium polykrikoides) against human immunodeficiency virus and other enveloped viruses [J]. International Journal of Biological Macromolecules, 1995, 17(5): 293-297.
[29] CARLUCCI M J, CIANCIA M, MATULEWICZ M C, et al. Antiherpetic activity and mode of action of matural carrageenans of diverse structural types [J]. Antiviral Research, 1999, 43(2):93-102.
[30] DAMONTE E, NEYTS J, PUJOL C A, et al. Antiviral activity of a sulphated polysaccharide from the red seaweed Nothogenia fastigiata [J]. Biochemical Pharmacology, 1994, 47(12): 2187-2192.
[31] PUJOL C A, COTO C E, DAMONTE E B.Determination of the antiviral activity of a naturally occurring sulftated xylomannan under various experimental conditions [J]. Rev Agent Microbiol, 1995, 27(2):91-98.
[32] CRANCE J M, GRATIER D, GUIMT J, et al. Inhibition of sacdfly fever Sicilian virus (phlebovirus) replication in vitro by antiviral compounds [J]. Res Virot, 1997, 148(5):353-365.
[33]王长云,管华诗.多糖抗病毒作用研究进展Ⅲ.卡拉胶及其抗病毒作用[J].生物工程进展,2000a,20(3):39-42.
[34] SUTAPA M, PRODYUT K, GHOSAL G A, et al. Isolation, chemical investigation and antiviral activity of polysaccharides from Gracilaria corticata (Gracilariaceae, Rhodophyta) [J]. International Journal of Biological Macromolecules, 2002, 31(1-3):87-95.
[35] HOSHINO T, HAYASHI T, HAYASHI K, et al. An antivirally active sulfated polysaccharide from Sargassum horneri(TURNER) C. AGARDH [J]. Biological & pharmaceutical bulletin, 1998, 21(7):730-734.
[36] PREEPRAME S, HAYASHI K, LEE JB, et al. A novel antiviraliy active fucan sulfate derived from an edible brown alga, Sargassum horneri [J]. Chemical & pharmaceutical bulletin (Tokyo), 2001, 49(4):484-485.
[37] REN D L, WANG J Z, AMANO H, et al. The effects of an algal polysaccharide from Gloiopeltis tenax on transplantable tumors and immune activities in mice [J]. Phanta Med, 1995, 61(2):120-124.
[38] COOBE D. Antitumor activity of ediblemmarine algae: effect of crude fucoid an fraction prepared from edible seaweeds against tumor [J]. International Journal of Cancer, 1987, (39):82-85.
[39] ITOCH H, NODA H, AMANO H, et al. Immunological analysis of inhibition of lung metastases by fucoidan (GIV-A) prepared from brown seaweed Sargassum thunbergii [J]. Anticancer Research, 1995, 15(5B): 1937-1947.
[40] ZHUANG C, ITOH H, MIZUNO T, et al. Antitumor active fucoidan from the brown seaweed, umitoranoo(Sargassum thunbergii) [J]. Biosci Biotechnol Biochem, 1995, 59(4):563-567.
[41]西泽一俊.海藻生物活性物质.食品开发(日),1980,18(11):28-30.
[42] OKAI Y, ISHIZAKA S, HIGASHI-OKAI K. Detection of immunomodulating activities in an extract of Japanese edible seaweed, Laminaria Japonica(Makonbu) [J]. Journal of the Science of Food and Agriculture, 1996, 72(4):455-460.
[43]董为人,田雪梅.补体活化在动脉粥样硬化形成中的意义[J].国外医学免疫分册,1999,22(1):47-50.
[44] KUZNETSOVA T A, ZAPOROZHETS T S, BESEDNOVA N N, et al. Immunostimulating and anticoagulating activity of fucoidan from browm algae Fucus evanescens of Okhotskoe sea [J]. Antbiot Khimioter, 2003, 48(4):11-13.
[45]张全斌,李大新.坛紫菜多糖对脾细胞活性的影响[J].中国海洋药物,2003,96(6):14-18.
[46]王常青,杨桂兰.两种海藻多糖对大鼠脂质代谢和血小板功能的比较[J].中华预防医学杂志,1997,31(6):342-345.
[47]王艳梅,李智恩,牛锡珍,等.孔石莼多糖降血脂活性的初步研究[J].中国海洋药物,2003,22(2):33-35.
[48]左绍远,钱金,万顺康,等.螺旋藻多糖降血糖活性实验研究[J].时珍国医国药,2000,11(8):677-678.
[49]王兵,李靖,马舒冰.羊栖菜多糖降血糖的实验研究[J].中国海洋药物,2000,(3):33-35.
[50]李八方,毛文君,曹立民.羊栖菜多糖对高血脂模型大鼠血脂的影响[J].中国水产科学,2000,7(2):56-58.
[51] CHURH F C, MEADE J B, TREANOR R E, et al. Antithrombin activity of fucoidan.The interaction of fucoidan with heparin cofactorⅡ. AntithrombinⅢ, and thrombin [J].The Journal of Biological Chemistry, 1989,264(6):3618-3623.
[52] TAKASHI N, AKIHIRO F, TERUKAZU N, et al. Inhibition of the generation of thrombin and factor Xaby a fucoindan from the brown seaweed Ecklonia kurome [J]. Thrombosis Research, 1999, 96(1): 37-49.
[53] GRAUFFEL, KLOAREG, MABEAU, et al. New natural polysaccharides with potent antithrombic activity: fucans from brown algae [J].Biomaterials, 1989, 10(6): 363-368.
[54] COLLIEC, FISCHER, TAPON-BRETAUDIERE, et al. Anticoagulant properties of a fucoidan fraction [J]. Thrombosis Research. 1991, 64(2): 143-154.
[55] MATSUBARA, BACIC. Anticoagulant properties of a sulfated galactan preparation from a marine green alga Codium cylindricum [J]. International Journal of Biological Mzcromolecules, 2001, 28(5): 395-399.
[56]王春玲,张全斌.褐藻多糖硫酸酯抗凝血活性的研究[J].中国海洋药物,2005,24(5):36-38.
[57] MARIANA S. P, ANA-CRISTINA E. S. VILELA-SILVA, ANA-PAULA VALENTE, et al. A 2-sulfated, 3-linked-α-Lgalactan is an anticoagulant polysaccharide [J]. Carbohydrate Research. 2002, 337(21-23), 2231–2238.
[58]王静凤,张学成,姜国良,等.枝管藻多糖的提取及其抗凝血活性的初步研究[J].青岛海洋大学学报,2003,33(1):75-79.
[59] Hayakawa Y , Hayashi T, Lee J, et al. Inhibition of thrombin by sulfated polysaccharides isolated from green algae[J]. Biochimaica et Biophysica Acta, 2000, 1543(1):86-94.
[60] Yasantha A, Kil-Nam K, You-Jin J. Antiproliferative and antioxidant properties of an enzymatic hydrolysate from browm alga, Ecklonia cava [J]. Food and Chemical Toxicology, 2006, 44(7):1065-1074.
[61] Hu, J. F., Geng, M.Y., Zhang, J. T., Jiang, H. D.. An in vitro study of the structure-activity relations of sulfated polysaccharide from brown algae to its antioxidant effect [J]. Journal of Asian Natural Product Reports, 2001, 3(4): 353-358.
[62] Zhang Q., Li, N., Zhou, G., et al. In vivo antioxidant activity of polysaccharide fraction from Porphyra haitanesis (Rhodephyta) in aging mice [J]. Pharmacological Research, 2003,48(2): 151-155.
[63] Zhang Q, Yu P, Li Z, et al,. Antioxidant activities of sulfated polysaccharide fractions from Porphyra haitanesis[J]. Journal of Applied Phycology , 2003,15(4):305-310.
[64]项华,赵健忠,柏林.海藻多糖抗突变性研究[J].癌变·畸变·突变,2001,13(1):42-44.
[65]陈美珍,余杰,龙梓洁,等.龙须菜多糖抗突变和清除自由基作用的研究[J].食品科学,2005,26(7):219-222.
[66]余杰,许肇成,颜璐璐,等.海萝多糖抗突变与抗肿瘤作用的研究[J].汕头大学学报,2007,22(2):59-63.
[67]庞辉,汤桂芳,玉艳红.螺旋藻多糖的抗溃疡作用及机制[J].广西医学,2006,28(11):1772-1773.
[68] Maisin JR. Bacq and Alecander Award lecture-chemical radioprotection: past, resent, and future prospects[J]. Int J Radiat Biol, 1998, 73(4): 443.
[69]吴晓旻,杨明亮,黄晓兰.海带多糖的抗辐射作用与脾细胞调亡[J].武汉大学学报,2004,25(3):239-241.
[70]罗琼,吴晓旻,杨明亮,等.海带多糖的抗辐射作用与淋巴细胞凋亡关系研究[J].营养学报,2004,26(12):471-473.
[71] Crist ,R. H., K. Oberholser, N. Shank, et al. Nature of Bonding Between Metallic Ions and Algal Cell Walls [J]. Environ. Science & Technol, 1981, 15(10):1212-1217.
[72] Tarlan E, Yetis U, Dilek F B. Algal treatment of pulp and paper industry wastewasters in SBR systems[J]. Water Science and Technology, 2002, 45(12):151-158.
[73] SAEKI Y, KATO T, OKUDA K. Inhibitory effects of funoran on the adherence and colonization of oral bacteria[J]. Bullentin of Tokyo Dental College, 1996, 37(2):77-92.
[74] MATSUI M S, MUIZZUDDIN N, ARAD S, et al. Sulfated polysaccharides from red microalgae have anti-inflammatory properties in vitro and in vivo [J]. Appl Biochem Biotechnol, 2003, 104(1):13-32.
[75] HINCHA D K, ZUTHER E, HELLWEGE E M, et al. Specific effects fructo-and glucooligosaccharides in the preservation of liposomes during drying [J]. Glycobiology, 2002, 12(2):103-110.
[76] EHRESMANN D W, DEIG E F, HATCH M T, et al. Dextran sulfate, a potent anti-HIV agent in vitro having synergism with sidovudine [J]. Lancet, 1987, 13(1):1379-1386.
[77] NARDELLA A, CHAUBET F, BOISSON-VIDAL, et al. Anticoagulant low molecular weight fucans produced by radical process and ion exchange chromatography of high molecular weight fucans extracted from the brown seaweed Ascophyllum nodosum [J]. Carbohydrate Research, 1996, 289(8): 201-208.
[78] YASANTHA A, WON-KYO J, THAVA V, et al. An anticoagulative polysaccharide from an enzymatic hydrolysate of Ecklonia cava [J]. Carbohydrate Polymers ,2006, 66(2): 184-191.
[79] SHANMUGAM S, MODY K H. Heparonid active sulfated polysaccharides from marine algae as potential blood coagulant agents [J]. Currant Sciences, 2000, (79): 1672-1683.
[80] SUSANNE A, GERHARD F. Characterization of the Anticoagulant Actions of a Semisynthetic Curdlan Sulfate[J]. Thrombosis Research, 2000, 99 (4): 377-388.
[81]刘志峰,宫晓黎,魏淑贞.五种海藻多糖体外抗血小板聚集作用观察[J].中国海洋药物,2001,20(2):36-38.
[82] TSIPALI E, WHALEY S, KALBFLEISCH J, et al. Glucans exhibit weak antioxidant activity, but stimulate macrophage free radical activity [J]. Free Radical Bicology and Medicine, 2001, 30(4):393-402.
[83] GROTH T, WAGENKNECH T. Anticoagulant potential of regioseselective Derivatized cellulose [J]. Biomaterials, 2001, 22(8): 2719-2729.
[84] MARIA G P, NORMA M B B, MARCI R S, et al. Structure and anticoagulant activity of a sulfated galactan from the red alga, Gelidium crinale. Is there a specific structural requirement for the anticoagulant action? [J]. Carbohydrate Research, 2005, 340(12): 2015-2023.
[85] MULLOY B, MOURAO PAS, GRAY E, et al. Structure/function studies of anticoagulant sulphated polysaccharides using NMR [J]. Journal of biotechnology, 2000, 77(1): 123-135.
[86]诸葛健,赵振锋,方慧英.功能性多糖的构效关系[J].无锡轻工大学学报,2002,21(2):209-212.
[87]中国农业百科全书,水产业卷[M].北京:中国农业出版社,1994,218-219,748-750.
[88]曾呈奎.中国经济海藻志[M].北京:科学出版社,1962:158-212.
[89]黄文凤,黄建明,董飞强.红毛菜的营养成分特征和价值[J].海洋水产研究,1998,19(2):57-61.
[90]马家海,李水军,纪焕红,等.红毛菜的氨基酸和脂肪酸分析[J].中国海洋药物杂志,2002,5:40-42.
[91]食品中蛋白质的测定方法.中华人民共和国国家标准,GB/T 14771-1993.
[92]黄伟坤.食品检测与分析[M].北京:轻工业出版社.1997:49-51.
[93]夏敏.必需微量元素与人体健康[J].中国食物与营养,2003,(10):53-56.
[94]藻类制品卫生标准,GB-19643-2005.
[95]王进卿.膳食纤维的功能与应用[J].中国酿造,2003,(5):37-38.
[96] DODGSON K S, PRICE R G. A note on the determination of the ester sulfate content of sulfated polysaccharides[J]. Biochemistry Journal, 1962, 84(1): 106-110.
[97]李宏燕,樊君.大枣多糖的水提醇沉工艺研究[J].宁夏工程枝术,2005,4(3):265-267.
[98]刑小黑,吴明忠,朱述钧,等.灵芝多糖化学研究[J].中国食用菌,1996,15(3):1-16.
[99]刘晓红,肖凯军.金樱子粗多糖的脱蛋白研究[J].食品科技,2007,(1):192-194.
[100]周存山,马海乐,胡文彬.条斑紫菜多糖提取工艺的优化[J].农业工程学报,2006,22(9):194-197.
[101]余华.海带多糖中蛋白质去除方法的对比研究[J].成都大学学报,2005,24(4):265-268.
[102]李瑞军,李德耀,张现峰,等.大孔吸附树脂法去除淫羊藿多糖中蛋白的研究[J].高等学校化学学报,2006,27(1):60-70.
[103]贾建波,李相前,杨文.桑黄多糖分离纯化及其结构初步鉴定[J].食品科学,2006,27(12):446-450.
[104]黄纯,马驰,宋慧智.亮菌多糖提取中脱蛋白和脱色方法比较[J].药学与临床研究,2007,15(1):45-46.
[105]陈小萍,张卫明,史劲松,等.茶树花水提多糖的精制工艺初探[J].食品科技,2007,32(4):72-75.
[106]何传波,陈玲,李琳.巴戟天多糖脱蛋白方法的研究[J].食品科技,2005(6):25-27.
[107]朱美静,童群义.猴头多糖脱蛋白方法的研究[J].河南工业大学学报,2005,26(4):25-27.
[108]盛家荣,曾令辉,翟春,等.多糖的提取、分离及结构分析[J].广西师院学报(自然科学版),1999,16(4):49-54.
[109]陈京,熊耀康.多糖类药物的质量标准研究现状及其进展[J].浙江中医学院学报,2004,28(3):77-80.
[110]张丽萍,李森,黄丽萍.金顶侧耳酸提水溶性多糖的研究——PC-3的分离、纯化与结构确定[J].菌物学报,1993,12(2):158-162.
[111]康琰琰,王一飞,朱良,等.裙带菜茎中硫酸多糖的分离纯化及分析鉴定[J].中药材,2005,28(9):769-771.
[112]黄益丽,郑天凌.海洋生物活性多糖的研究现状与展望[J].海洋科学,2004,28(4):58-61.
[113]张承圭,王传怀,袁玉荪,等.生物化学仪器分析及技术[M].北京:高等教育出版社,1990:84-90.
[114]刘成梅,付桂明,游海,等.百合多糖的纯化与化学结构鉴定研究[J].食品科学,2002,23(5):114-117.
[115]马丽,覃小林,刘雄民,等.螺旋藻多糖的纯化和效液相色谱分析[J].化工技术与开发,2004,33(2):29-32.
[116]董洪新,刘新海,李焕艺.灵芝多糖生物活性物质的研究现状及进展[J].当代生态农业,2003,(1):28-30.
[117] XIN X L, LIU C H. Progress in the study of antioxidant effects of Chinese herbal polysaccharides [J]. Journal of Beijing University of Traditional Chinese Medicine, 2000, 23(5): 54-55.
[118] HALLIWELL B, GUTTEIDE J M C, ARUMOA O I. The deoxyribose method: A simple“Test-Tube”assay for determination of rate constants for reactions of hydroxyl radicals [J]. Analytical Biochemistry, 1987, 165(1): 215-219.
[119]吉林市卫生防疫站.GB/T 5009.171-2003保健食品中超氧化物歧化酶(SOD)活性的测定[S].中国国家标准化管理委员会.北京:中国标准出版社,2003:410-417.
[120] DUAN X J, ZHANG W W, LI X M, et al. Evaluation of antioxidant property of extract and fractions obtained from a red alga, Polysiphonia urceolata [J], Food Chemistry, 2006, 95(1): 37 .
[121]范晓,严小军.高分子量褐藻多酚抗氧化性质研究[J].水生生物学报,1999,23(5):494-499.
[122] YI O S, MEYER A S, FRANKED E N. Antioxidant Activity of Grape Extracts in a Lecithin Liposeme System[J]. JAOCS, 1997, 74(10): 130l-1306.
[123]晏文洁,李家璞.类黄酮抗氧化力与其结构之关系[J].台湾农业化学与食品科学,2000,38(1):80-88.
[124] CUZZOCREA S, RILEY D, CAPUTI A P, et al. Antioxidant therapy: a new pharmacological approach in shock, inflammation and ischemis/reperfusion injury[J]. Pharmacological reviews, 2001, 25(5): 135-139.
[125]钦传光,周军,赵文,等.泥鳅多糖清除活性氧和保护DNA链的作用[J].生物化学与生物物理学报,2001,33(2):215-218.