蜜环菌多糖的分离纯化及PMP衍生化HPLC分析

目的:对蜜环菌发酵液进行多糖的提取、分离纯化,得到均一多糖,对其进行单糖组分分析.方法:分离纯化采用分步醇沉和葡聚糖凝胶(Sephadex G-200)色谱法;纯度鉴定及分子量测定采用高效液相色谱法;单糖组成采用1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生HPLC法测定.结果:获得两种均一多糖(AMFP-Ⅰ、AMFP-Ⅱ),峰位分子量分别为 812 939D、596 217D,重均分子量(MW)分别为995 098D、872 640D,分布宽度(Mw/Mn)分别为1.268 93、1.235 20;测定单糖组成为甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖.结论:两种多糖分子量分布及单糖组成均不同,AMFP-Ⅰ主要由半乳糖醛酸、葡萄糖、半乳糖组成,AMFP-Ⅱ主要由半乳糖醛酸、半乳糖、木糖组成.     

三百棒多糖的单糖组成分析及对巨噬细胞泡沫化的作用研究

目的 研究三百棒多糖（Toddalia asiatica Lam polysaccharides，TALP）的单糖组成及TALP对巨噬细胞泡沫化形成的影响，初步探讨TALP抗动脉粥样硬化的可能机制。方法 采用水提醇沉法提取多糖，除蛋白后干燥，得粗多糖，利用Sephadex-G200纯化三百棒粗多糖，得TALP组分1（TALP1）和组分2（TALP2），1-苯基-3-甲基-5-吡唑啉酮柱前衍生高效液相色谱（PMP-HPLC）法和红外光谱法测定TALP组分和结构；将人外周血单核细胞系（THP-1）用十四烷酸佛波醇酯（PMA）诱导分化为巨噬细胞后分为对照组、氧化低密度脂蛋白（Ox-LDL）组及Ox-LDL+TALP1组，然后油红O染色并分析各组细胞油红O染色着色面积。结果 TALP红外光谱结果呈现多糖的典型特征吸收峰，TALP1是由葡萄糖、甘露糖、阿拉伯糖、鼠李糖、半乳糖、葡萄糖醛酸、半乳糖醛酸、果糖及另外2种未知的单糖构成的一种水溶性杂多糖。油红O染色结果显示，对照组基本无油红O着色，Ox-LDL组油红O着色面积明显增加，经过TALP1干预以后，Ox-LDL+TALP1组油红O着色面积减少。结论 TALP是一种杂多糖，含有葡萄糖、甘露糖、阿拉伯糖、鼠李糖、半乳糖、葡萄糖醛酸、半乳糖醛酸、果糖等；TALP1可抑制巨噬细胞泡沫化。     

银杏蜜环口服溶液中单糖HPLC特征图谱及其质量评价

目的 利用HPLC建立银杏蜜环口服溶液中糖的特征图谱，并对不同来源的银杏蜜环口服溶液质量进行评价。方法 采用1-苯基-3-甲基-5-吡唑啉酮（1-phenyl-3-methyl-5-pyrazolone，PMP）柱前衍生HPLC，结合中药指纹图谱相似度分析软件对4批次市售银杏蜜环口服溶液和6批次自制口服溶液中单糖进行相似度分析，同时通过对组方中成分单糖组成进行测定，对共有峰进行了归属和确认。结果 通过对自制口服溶液与市售口服溶液的HPLC特征图谱进行研究，确定了5个共有峰，经过归属验证，分别为甘露糖、鼠李糖、葡萄糖、半乳糖和阿拉伯糖，相似度介于0.998~1.000之间；同时对口服溶液及其组方中原药材进行了单糖组成测定，发现水解口服液中单糖来源于蜜环菌粉、银杏叶提取物及矫味剂甜菊糖；进一步对不同厂家的银杏叶提取物和蜜环菌粉进行单糖组成测定，结果显示不同厂家的银杏叶提取物和蜜环菌粉单糖组成有明显差异。最后通过对口服液及其原辅料中多糖进行提取、测定，发现口服液中多糖主要来源于蜜环菌粉。结论 本研究建立的方法能较全面地体现银杏蜜环口服溶液组分中单糖的特征，将市售口服溶液、自制口服溶液及其原药材的水解单糖进行归属及溯源，可用于口服液及其原料药材、辅料的质量评价。     

西洋参多糖PPQI-1～4理化性质的研究

AIM: To study the characteristics of the four bioactive pure polysaccharides of Panax quiquefolium L. (PPQI 1～4). METHODS: The polysaccharides were extracted with hot water and grade precipitated with ethanol. They were further purified with Sephadex DEAE A50 and Sephadex G200 chromatography. The molecular weights of the polysaccharides were determined with matrix assisted laser desorption/ionization mass spectrometry (MALDI MS). The sugar compositions were analyzed with GC. The linkage positions of the component sugars were determined by methylation and GC MS. RESULTS: The four polysaccharides are all uronic acid containing heteropolysaccharides with the component sugars of arabinose, galactose and glucose in different molar ratios. The uronic acid contents are within 30%～60%. Their molecular weights are in the range of 2×10⁴～7×10⁴u. CONCLUSION: The four new bioactive polysaccharides are all uronic acid containing heteropolysaccharides.     

FT-IR study of the polysaccharides isolated from the skin juice, gel juice, and flower of Aloe vera tissues affected by fertilizer treatment

Background

This experiment was conducted to evaluate the effect of different amounts of fertilizers on the polysaccharides of Aloe vera plant. There were four different treatments, viz. T₁?=?150%?N, T₂?=?150% P, T₃?=?150%?K, and T₄?=?150% NPK (50%?N?+?50% P?+?50%?K) soil. Crude water-soluble polysaccharides were isolated from the gel juice, skin juice, and flowers of A. vera planted in these soils.

Results

Result indicates that skin juice contained 2.4 times the level of polysaccharides in gel juice from one plant, suggesting the potential industrial application of A. vera skin rather than discarding it. After anion-exchange chromatography, neutral polysaccharides accounted for 58.1% and 78.5% of the total recovered neutral and acidic polysaccharide preparations from the gel juice and skin juice, respectively, whereas the crude flower polysaccharides were largely composed of weakly acidic polysaccharides (84.2%). Sugar analysis of the polysaccharides after gel permeation chromatography revealed that glucose and galactose were the most abundant monosaccharide in the neutral polysaccharides from the gel juice and skin juice, respectively. The acidic polysaccharides from the two juices consisted of glucuronic acid, galactose, glucose, mannose, and xylose with variable proportions.

Conclusions

Except glucuronic acid (15.4%) in flower acidic polysaccharide, the flower neutral and acidic polysaccharides contained galactose, glucose, and mannose as the main sugar components. Glucuronic acid was the major uronic acid in all acidic polysaccharides from different tissues.     

新疆药桑叶、枝多糖中单糖的组成分析与测定

目的:分析新疆药桑叶、枝多糖中单糖的组成并对其进行测定。方法:采用水提醇沉法提取新疆药桑叶、枝中的多糖,用2mol·L-1H2SO4将多糖水解成单糖,以1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化;采用反相高效液相色谱法测定各单糖。结果:桑叶、桑枝多糖均是由甘露糖、鼠李糖、葡糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖等单糖组成。7种单糖在25min内可很好地分离。结论:本方法操作简便、灵敏度高、结果准确,可用于新疆药桑的质量控制。     

盐碱地植物内生真菌SXH-69胞外多糖的分离、化学组成和结构特征

目的 对分离自滨州盐碱地植物内生真菌SXH-69的胞外多糖进行研究。 方法 利用乙醇沉淀法? Q Sepharose Fast Flow离子交换柱色谱和Sephacryl S-100凝胶柱色谱对菌株发酵液中的胞外多糖进行分离纯化;通过高效凝胶渗透色谱(HPGPC)? PMP柱前衍生高效液相色谱(HPLC)? 红外光谱(IR)及气质联用色谱(GC-MS)等方法,研究胞外多糖的化学组成和结构特征。 结果 从发酵液中分离纯化获得2个胞外多糖sx1-1和sx2-1,它们的分子量分别为13.5 kDa和18.3 kDa;这2个多糖主要由甘露糖组成,还含少量葡萄糖和半乳糖;sx1-1和sx2-1中均含有末端Man,1,4)-Man,1,6)-Man和1,2,4)-Man,而sx1-1还含有末端Glc, 1,2)-Gal,1,6)-Glc,1,3,6)-Glc;sx2-1中还含有1,2)-Glc,1,2,6)-Gal。 结论 首次从滨州盐碱地植物内生真菌SXH-69中分离得到胞外多糖sx1-1和sx2-1,它们有着不同的化学特征,是结构新颖的杂多糖。     

灵芝多糖分离鉴定及抗肿瘤活性的研究

目的探讨灵芝多糖的分离纯化、理化性质及初步抗肿瘤活性。方法采用热水提取 ,乙醇分级沉淀 ,DEAE 32离子交换柱色谱等方法从灵芝子实体中分离得到灵芝多糖。用SephadexG 10 0凝胶柱色谱法测定其纯度和平均分子量。用薄层色谱及气相色谱法测定其单糖组成。溴化四唑蓝法测定其初步体外抗肿瘤活性。结果从灵芝子实体中分离得到 4种均一多糖 (GLPL1 ～GLPL4 ) ,其中GLPL1 和GLPL3为主要成分 ,其分子量为 4 10 0和 5 70 0。GLPL1 由葡萄糖组成 ;GLPL3由葡萄糖和半乳糖组成。GLPL1 和GLPL3对人鼻咽癌细胞增殖有显著的抑制作用 ,GLPL3还对人胃癌细胞、人结肠癌细胞增殖有一定的抑制作用。结论灵芝多糖GLPL1 、GLPL3是很有潜力的抗肿瘤或辅助抗肿瘤药物     

人参细胞培养物中水溶性多糖的研究

人参细胞培养物中水溶性多糖经提取后,用果胶酶和Sevag法除去其中蛋白,用Sephadex G-100柱进行分离、纯化得到DPSCG1和DPSCG2两个洗脱峰。经醋酸纤维素薄膜电泳和琼脂糖凝胶电泳,证明DPSCG1和DPSCG2均为均一性多糖,两者经水解和薄层层析结果均由鼠李糖、木糖、阿拉伯糖、半乳糖和半乳糖醛酸等组成,经Sephadex G-200柱层析并与标准分子量对比,DPSCG1和DPSCG2的分子量分别为1.22×10⁵D和2.79×10⁴D。     

Immunostimulating Polysaccharides from Panax notoginseng

Purpose. The main purpose of this study is to prepare and characterize polysaccharides from Panax notoginseng, investigate their effects on immune system in vitro in order to find new immunostimulants for the prevention and supporting treatment of infection and immunodeficiency related diseases. Methods. Polysaccharides were extracted with aqueous solution, separated with column chromatography. Their anticomplementary activities were investigated by using human serum and antibody-sensitized sheep red blood cells. Interferon- and tumor necrosis factor inductive activities were studied by using isolated mouse spleen lymphocytes and peritoneal macrophages, respectively. Results. Four polysaccharides, homogeneous in gel-filtration chromatography, were prepared and designated PF3111, PF3112, PBGA11, and PBGA12. Component sugar analysis revealed that they are heteroglycans with MWs ranging from 37 kD to 760 kD, composed of glucose, galactose, arabinose, mannose, and xylose in different molar ratios. Fraction PBGA12 has the most anticomplementary activity which is mediated through both alternative and classical pathways. All the polysaccharides except PBGA11 induced the production of interferon- in the presence of concanavalin A. They induced the production of significant amount of TNF- in cell cultures. Conclusions. The polysaccharides from P. notoginseng have immunostimulating activities in vitro.     

Studies on antitumor and immunopotentiating activities of polysaccharides from Trichosanthes rhizome

The polysaccharide fraction from the rhizome ofTrichosanthes kirilowii (Cucurbitaceae) showed marked antitumor and cytotoxic activity with immunopotentiating activity. It was evidenced by the increase in the number of circulating leucocytes and peritoneal exudate cells and the recovery of lowered antibody forming activity in mice. The polysaccharide was mainly composed of glucose, galactose, fructose, mannose and xylose and a samll amount of protein.     

榆耳多糖的分离纯化、结构鉴定及抗肿瘤活性研究

目的对榆耳粗多糖(GI)进行分离纯化,并对所得均一多糖进行结构鉴定及抗肿瘤活性的研究。方法 GI经Q-Sepharose F.F离子交换色谱,Sepharose CL-6B凝胶色谱分离得到均一多糖,采用凝胶色谱、薄层色谱、红外光谱、高碘酸氧化等方法测定相对分子质量(Mr)、单糖组成、糖苷键连接方式。采用MTT法对粗多糖和均一多糖的体外抗肿瘤活性进行测定。结果分离得到均一多糖GI1-3和GI1-4b,其Mr分别约为1.55×105和3.03×104。经薄层色谱法、红外光谱法、高碘酸氧化法确定GI1-3由半乳糖、葡萄糖、甘露糖、木糖组成,以β-(1,4)、β-(1,6)、β-(1,3)糖苷键连接。GI1-4b由葡萄糖、甘露糖和木糖组成,以β-(1,4)、β-(1,6)糖苷键连接。体外抗肿瘤实验表明,GI1-3的抑瘤活性较GI1-4b和GI高,GI的抑瘤活性在3种多糖组分中最低。结论 GI、GI1-3和GI1-4b对体外生长的肿瘤细胞具有不同的抑制作用,可能与其Mr及分子结构有关。     

The isolation and the characterization of two polysaccharides from the branch bark of mulberry (<Emphasis Type="Italic">Morus alba</Emphasis> L.)

Two water-soluble polysaccharides termed MBBP-1 and MBBP-2 were isolated from the branches of the mulberry tree (Morus alba L.) using hot water extraction and purified on Anion-exchange DEAE52-cellulose and Sephadex G-100 column. MBBP-1 was shown to be composed of rhamnose, xylose, arabinose, mannose, glucose and galactose in the molar ratio of 4.53:2.49:4.38:4.67:17.85:5.88. MBBP-2 was composed of rhamnose, xylose, arabinose, mannose, glucose, galactose and galacturonic acid in the molar ratio of 26.85:13.8:3.14:4.4:6.1:3.19:4.9. Their structural characteristics were further investigated by FI-IR spectroscopy, Smith degradation, methylation analysis and NMR spectroscopy. Based on the data obtained, MBBP-1 had a backbone mainly consisting of (1 → 3)-linked glucose. MBBP-2 had a backbone mainly consisting of (1 → 3)-linked rhamnose and (1 → 2, 4)-linked xylose. Antioxidant assays indicated that antioxidant activities of MBBP-2 were significantly stronger than those of MBBP-1, and this was likely in relation to the different content of 8.2 % galacturonic acid in MBBP-2.     

灵芝多糖的提纯、组成及活性研究

目的探讨灵芝多糖的分离纯化、单糖组成以及抗肿瘤活性。方法采用热水提取,Sevag法去蛋白质,乙醇分级沉淀,DEAE-32离子交换色谱法从灵芝子实体粉中分离得到灵芝多糖。用SephadexG-100凝胶柱色谱法判断其纯度及相对分子质量,用薄层色谱法及气相色谱法鉴定其单糖组成,溴化四唑蓝法测定其体外抗肿瘤活性。结果从灵芝子实体粉中分离得到两种粗多糖GLPⅠ、GLPⅡ。GLPⅠ进一步分离得到两种均一多糖GLPH1、GLPH2。GLPH1糖苷键为β型,相对分子质量为45000。GLPH1由葡萄糖、半乳糖和痕量甘露糖、岩藻糖组成。GLPH1对KB细胞、BGC细胞、B16细胞的增殖具有一定的抑制作用。结论灵芝多糖GLPH1可望开发成抗肿瘤药或辅助抗肿瘤药。     

山药多糖的提取分离及山药总多糖的含量测定

目的:提取分离并测定山药多糖的含量。方法:依次用水提取,乙醇和十六烷基三甲基溴胺盐沉淀法得到粗多糖;用葡聚糖凝胶柱层析和高效液相层析纯化多糖并测定分子量;采用HPLC法测定山药总多糖含量。结果:从山药中得到两个均一多糖,相对分子质量分别为62000和7300Dal;HPLC法测得山药总多糖含量为16.42%。结论:采用本实验提取方法可有效提取分离出纯化山药多糖,利用HPLC法测定山药总多糖含量结果与其他测定方法比较,结果准确,可作为山药总多糖含量测定的方法。     

柱前衍生化HPLC法测定黄河滩枣多糖的单糖组成

目的建立PMP(1-苯基-3-甲基-5-吡唑啉酮)柱前衍生化HPLC法测定黄河滩枣多糖的单糖组成。方法采用水提醇沉法提取黄河滩枣多糖,多糖水解后进行PMP衍生化,HPLC法测定黄河滩枣多糖的单糖组成及摩尔比,并对等度和梯度洗脱两种模式的测定结果进行分析比较。结果黄河滩枣多糖由D-甘露糖、L-鼠李糖、D-半乳糖醛酸、D-葡萄糖、D-半乳糖、L-阿拉伯糖等六种单糖组成,它们的摩尔比约为0.54∶0.50∶0.27∶2.03∶5.40∶1.00,等度和梯度洗脱测定结果基本一致。结论本实验改进的等度洗脱方式更加简便易行,快速准确,适用于黄河滩枣多糖的单糖组成分析。     

Composition and Antiinflammatory Activity of Polysaccharide Complexes Extracted from Sweet Violet and Low Mallow

Water-soluble polysaccharides extracted from the grass of sweet violet (Viola odorata L.) and low mallow (Malva pusilla Smith) include glucose, galactose, arabinose, rhamnose, xylose, and glucuronic and galacturonic acids. The results of tests on experimental animals showed that the polysaccharide complexes from both medicinal plants studied possess antiinflammatory activity, which is manifested by suppression of the exudation and proliferation stages of inflammation and by a change in the capillary permeability. __________ Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 39, No. 4, pp. 29 – 32, April, 2005.     

蛹虫草菌丝体多糖与子实体多糖体外免疫调节活性的研究

目的 分析蛹虫草菌丝体多糖（mycelia polysaccharides of Cordyceps militaris,MPS）与蛹虫草子实体多糖（polysaccharides from fruiting body of Cordyceps militaris,FPS）的体外免疫活性。方法 以RAW264.7细胞株为受试对象,从细胞增殖、中性红吞噬能力、NO释放水平角度系统考察不同浓度的MPS与FPS的免疫调节活性,并分析其量化关系。结果 MPS与FPS均可明显促进RAW264.7细胞株的增殖、对中性红的吞噬能力和NO释放水平,且均呈现出浓度依赖性的特征。二者对RAW264.7细胞株增殖的促进作用无显著性差异,但在免疫调节最佳作用浓度方面却存在一定差异,MPS在100μg·mL^-1时表现出最好的细胞吞噬能力,而FPS在200μg·mL^-1时表现出最高的细胞吞噬能力。结论 MPS与FPS均具有显著的体外免疫活性。     

蒲公英根多糖的分离、纯化及结构的初步分析

目的提取蒲公英根多糖,并对该多糖进行分离、纯化和结构的初步分析。方法采用超声波协同酶法进行提取多糖,分离纯化后,经Sephadex G-75柱分成A,B,C,D4级,采用优化的PMP柱前衍生HPLC分析单糖组成。采用紫外、红外等方法进行结构的初步分析。结果 4种多糖均含有D-鼠李糖、葡萄糖、D-半乳糖、D-木糖和D-阿拉伯糖;它们的单糖的摩尔比依次为,A：0.098∶2.88∶0.125∶0.01∶0.33;B：0.113∶0.68∶0.13∶0.011∶0.19;C：0.139∶2.46∶0.36∶0.13∶0.086;D：0.028∶0.36∶0.031∶0.012∶0.94。结论 4种多糖在结构上有明显的不同。     

Growth inhibitory activities of kalopanaxsaponins A and I against human pathogenic fungi

Antifungal activities of the compounds isolated fromKalopanax pictus against representative fungi of dermatomycosis were investigated using paper disc diffusion method. It was found that kalopanaxsaponins A and I were effective in inhibiting the growth ofCandida albicans KCTC 1940 andCryptococcus neoformans KCTC 7224 with minimum inhibitory concentration (MIC) of 25 μg/ml. It showed that antifungal activity of both compounds have strong selectivity against the fungi of dermatomycosis.