红芪多糖的提取分离纯化及组成分析

目的获得红芪多糖纯品并且得到其单糖组成。方法通过分步醇沉法和凝胶柱色谱获得红芪多糖纯品,用气相色谱法测其组成。结果红芪多糖1和2经Sephadex G-200纯化后分别得到两个组分,红芪多糖3通过Sephadex G-25得到两个组分;红芪多糖1,2,3均由鼠李糖、阿拉伯糖、木糖、葡萄糖、半乳糖五种单糖组成。结论分步醇沉法获得的三种多糖1,2,3经Sephadex G-200和Sephadex G-25纯化可获得红芪多糖纯品,GC法测其组成方法简便可行。     

黄芪多糖的研究

从中药黄芪Aslragalus mongholicus Bunge.的水提液中分离得到两种葡聚糖AG-1、AG-2和两种杂多糖AH-1、AH-2。该四种多糖经电泳、凝胶柱层析等证实均为单一组分。AG-1为水溶性,经水解、过碘酸氧化、甲酸生成、Smith降解、乙酰解、红外、C-13核磁共振等确定为α(1→4)(1→6)葡聚糖、α(1→4)与口(1→6)甙键糖基的组成比例约为5:2。AG-2不溶于水,为α(1→4)葡聚糖。AH-1为水溶性酸性多糖,水解后纸层析检出含己糖醛酸(半乳糖醛酸和葡萄糖醛酸)、葡萄糖、鼠李糖、阿拉伯糖,其分子比值为1:0.04:0.02:0.01。AH-2为水溶性、水解后用纸层析和气相层析检出葡萄糖和阿拉伯糖,其分子比值为1:0.15。AG-1、AH-1具有某些免疫促进作用。     

红芪多糖的提取分离纯化及组成分析

目的 获得红芪多糖纯品并且得到其单糖组成.方法 通过分步醇沉法和凝胶柱色谱获得红芪多糖纯品,用气相色谱法测其组成.结果 红芪多糖1和2经Sephadex G-200纯化后分别得到两个组分,红芪多糖3通过Sephadex G-25得到两个组分;红芪多糖1,2,3均由鼠李糖、阿拉伯糖、木糖、葡萄糖、半乳糖五种单糖组成.结论 分步醇沉法获得的三种多糖1,2,3 Sephadex G-200 和Sephadex G-25纯化可获得红芪多糖纯品,GC法测其组成方法简便可行.     

坛紫菜多糖对脾细胞活性的影响

目的：坛紫菜多糖分级组分F1、F2和F3是从坛紫菜中通过热水提取及DEAE-纤维素层析得到的3个多糖组分。本文研究了坛紫菜多糖组分对大鼠脾细胞活性的影响。方法：采用噻唑蓝(MTT)比色方法研究坛紫菜多糖组分对大鼠脾细胞活性的影响。结果：Fl和F2对脾细胞的活性具有明显的抑制作用，F3对脾细胞活性也有一定的抑制作用。而且3种多糖组分能够显著抑制紫外线损伤的脾细胞的活性。结论：坛紫菜多糖对免疫细胞的活性具有明显的抑制作用，可能是一种潜在的免疫抑制剂。     

马齿苋多糖POPⅡ的分离纯化及理化性质研究

为研究马齿苋多糖组分的性质，采用水提醇沉方法从马齿苋全草中提取粗多糖，通过Sevage法脱蛋白质后，再经DEAE-纤维素柱层析和SephadexG-200葡聚糖凝胶柱色谱分离纯化得到马齿苋精多糖POPⅡ，由醋酸纤维薄膜电泳和SephadexG-200葡聚糖凝胶过滤法鉴定为均一性组分，化学定性反应和光谱鉴定表明POPⅡ不含蛋白质和核酸、不含酚类物质，但含有糖醛酸的非淀粉类多糖，其相对分子质量为12100。POPⅡ完全酸水解后经薄板层析、气相色谱分析（结合半乳糖醛酸含量），确定它由阿拉伯糖、半乳糖和半乳糖醛酸组成，其中阿拉伯糖：半乳糖的摩尔比为1．00：1．05，其组成中半乳糖醛酸含量为9．18％。红外光谱分析表明POPⅡ具有典型的多糖吸收峰，结构中存在β-型糖苷键。     

贻贝多糖对大鼠睾丸支持细胞增殖作用的研究

目的对贻贝多糖进行分离提取并就其对大鼠睾丸支持细胞的增殖作用进行了研究。方法通过热碱法提取贻贝多糖粗品,过DEAE-cellulose柱获得3个洗脱峰的半纯品,然后用MTT法对其进行了大鼠睾丸支持细胞增殖率测定。结果得到的贻贝多糖粗品呈白色粉末状,水溶性好;贻贝多糖半纯品3个洗脱峰均能提高大鼠睾丸支持细胞的增殖率,其中,第3峰和1mg·mL-1剂量的第2峰促细胞增殖作用明显。结论贻贝多糖能有效提高大鼠睾丸支持细胞活性,从而增强机体的免疫力、促进男性生殖细胞的发育。     

大黄多糖分离纯化及其功能的研究（I）

大黄多糖的分离纯化，通过甲醇回流，除去大黄中致泻的蒽醌类物质，按Sevag‘s方法去除蛋白质，获得大黄多糖粗制品，再分别经Sephadex G-50和G－100柱分别纯化，结果获得单一组分多糖纯品；建立四氧嘧啶诱导的糖水病小鼠模型；大黄多糖有明显（P＜0．01），降低血糖及提高血清胰岛素水平的功能。     

石花多糖抗辐射有效部位的提取分离及检测

目的:对具有抗辐射作用的石花多糖粗品进行提取、分离和纯度检测.方法:将梅花衣科植物石花(Parmelia tinctorum Despr)水提、醇沉后,经AB-8树脂脱色和三氯乙酸除蛋白,再用DE22纤维素柱分离,冷冻干燥后得浅灰色粉末多糖;对石花多糖有效组分进行纸层析分析和玻璃纤维纸电泳纯度检查.结果:纯度检查不多于3个斑点.结论:石花多糖有效部位为相对单一组分.     

胀果甘草多糖的分离纯化及其理化性质

目的对新疆地产胀果甘草Glycyrrhiza inflataBat.中获得的一种水溶性多糖(GIP-2)进行分离纯化,并测定其部分理化参数。方法采用脱脂、回流提取、乙醇沉淀、Sevag法除蛋白,从胀果甘草药材中提取粗多糖,透析后经DEAE-52离子交换层析和Sepharose CL-6B、Sephadex G-50凝胶柱层析分离纯化得到一种水溶性多糖(GIP-2);UV及IR法检测其性质;自动旋光仪测定旋光度;高效凝胶渗透色谱法(HPGPC)分析其纯度和分子量范围;完全酸水解法鉴定多糖的单糖组分。结果胀果甘草多糖GIP-2为黄白色粉末,无甜味,易溶于水;UV检测192 nm处有明显吸收峰,260、280 nm处均无吸收峰,证明被测物为多糖,且不含核酸及蛋白质;IR分析结果显示,在3393、2932、1616、1423、1101 cm-1处表现为典型的多糖吸收峰;GIP-2分子量>2000 kDa,主要由葡萄糖、阿拉伯糖和半乳糖组成,其摩尔比为3.3:11.7:1.0。结论提取分离所得胀果甘草多糖GIP-2为单一、纯净的多糖。     

当归多糖X—C—3—Ⅱ的分离纯化与组成研究

目的对中药岷当归[Angelicasinensis(Oliv)Diels]的水溶性成分进行研究,分离出有活性的单一的多糖组分.方法采用热水提取、乙醇沉淀、DEAE-SephadexA-25柱层析进行分离,凝胶色谱法测定相对分子质量,利用气相色谱法鉴定多糖组分中所含单糖的种类和它们之间的摩尔比.结果分离得到一个多糖组分X-C3-Ⅱ,相对分子质量为1.0×105,其中所含单糖的种类和它们之间的摩尔比为葡萄糖半乳糖阿拉伯糖鼠李糖半乳糖醛酸=56.022.118.91.91.1.结论x-C-3-Ⅱ为首次从该植物中分离得到,并且为均一性多糖.     

Cell proliferative effect of polyxyloses extracted from the rhizomes of wild turmeric,Curcuma aromatica

Hot water-soluble crude polysaccharides were extracted from the rhizomes of wild turmeric, Curcuma aromatica Salisb. (Zingiberaceae), using dry grinding, boiling water extraction, and then ethanol precipitation. The crude polysaccharide extract was then fractionated by DEAE-cellulose ion exchange column chromatography, and subsequently further purified by Superdex G-200 gel filtration column chromatography, giving two relatively abundant polysaccharide fractions, called P11 and P21, and a much less common fraction P22 obtained in insufficient amounts for further analysis. The two main polysaccharide fractions were evaluated for monosaccharide composition by acid hydrolysis and high performance liquid chromatography (HPLC), whilst the molecular weight and functional groups were determined by gel permeable chromatography (GPC) and FT-IR, respectively. Fractions P11 and P21 were found to be polyxyloses with molecular weight-averages of 469,171 and 157,665?Da, respectively. P11 (100?μg/mL) could significantly induce human gingival fibroblast cells proliferation by 30%, while P21 (100?μg/mL) could significantly inhibit gingival fibroblast cells proliferation by 92%. The in vitro human primary gingival fibroblast cell proliferation in cell culture at a concentration of 100?μg/mL.     

Structural features of an immunostimulating and antioxidant acidic polysaccharide from the seeds of Cuscuta chinensis

Three crude polysaccharide fractions, named CS-A, CS-B and CS-C, were prepared from the seeds of Cuscuta chinensis by hot water extraction and diluted alkali extraction subsequently, then EtOH precipitation, and tested for lymphocyte proliferation activity. CS-A showed a stimulating effect on concanavalin A or lipopolysaccharide induced mitogenic activity of lymphocytes. An acidic polysaccharide (CS-A-3beta) was purified from CS-A by anion exchange and gel filtration chromatography. The polysaccharide showed potent stimulating effects on lymphocyte proliferation and antibody production, but did not significantly influence the serum IgG level. Its structural features were elucidated by methylation analysis, partial acid hydrolysis, 1D and 2D NMR and ESI-mass spectroscopy. The data obtained indicated that CS-A-3beta had a backbone consisting of alpha-D-1,4-linked GalpA residues and beta-L-1,2-linked Rhap residues with branches at C-4 of Rhap residues and C-3 of GalpA residues, composed of arabinogalactan and glucobiose. This structure is typical for a pectic polysaccharide of the rhamnogalacturonan type. In addition, the effect of CS-A, CS-B, CS-C and CS-A-3beta on hydrogen peroxide induced cell lesion in rat pheochromocytoma line PC 12 was investigated. The results indicated that, besides its immunostimulating activity, CS-A-3beta had a protective effect against free radical-induced cell toxicity.     

浒苔多糖的分离纯化及其对小鼠淋巴细胞活性的影响

为了探讨浒苔(Enteromorpha)多糖的免疫调节活性,采用Sevage法,Sephacryl-300 分子筛层析以及乙醇分级沉淀法获得四个浒苔多糖组分EP,EP1、EP2和EP3,比较四种多糖对体外培养的小鼠T、B淋巴细胞增殖影响;同时观察作用效果较强的EP2在T、B淋巴细胞中的分布及其对细胞RNA合成的影响。结果表明,EP、EP1、EP2和EP3中单糖、糖醛酸、硫酸根含量存在差异,均能促进对B淋巴细胞以及ConA或LPS诱导的T、B淋巴细胞增殖,且EP2的作用效果最好;经EP2处理的T、B淋巴细胞或同时用EP2和丝裂原(ConA或LPS)处理的T、B淋巴细胞体积明显增大,RNA合成增加,B淋巴细胞有母细胞化趋势。由此可见,浒苔多糖能调节小鼠淋巴细胞活性,促进丝裂原诱导的淋巴细胞分化和免疫应答,有望成为一种新的动物免疫增强剂。     

海带硫酸多糖的提取、纯化及其理化分析

目的研究海带硫酸多糖 (LPS)的提取制备及其组分的纯化和理化性质。方法采用酶解法从海带提取制备LPS,海带浆加入纤维素酶、半纤维素酶、果胶酶和蛋白酶 ,50℃水解 4h。取滤液加入氯化钙 ,离心去除海藻酸钙。取上清液加入十六烷基三甲基溴化铵 (CTAB)与LPS结合沉淀 ,离心收集CTAB LPS沉淀物。加入氯化钙溶液进行盐解 ,将LPS游离释放出来 ,加入乙醇使LPS析出。离子交换色谱和凝胶过滤色谱法分离纯化其多糖组分。结果酶解提取法的收率为 5‰ ,色谱法分离得到 4个主要多糖组分 ,测定其分子量分别为 2 1 0、1 2 0、40 0和 1 4 0kD。酶解法的提取收率高于水煮法。结论LPS的开发具有广阔的前景     

海胆黄多糖的分离、纯化及免疫活性测定

目的从光棘球海胆中分离纯化海胆黄多糖(polysaccharidefromtheeggsofStrongylocentrotusnu-dus,简称SEP),确定其纯度和分子量,并现察其免疫活性。方法海胆黄先经丙酮脱脂,根据正交实验和活性分析确定最佳热水提取条件,然后热水提取、去蛋白、醇沉得海胆黄粗多糖。粗多糖经超滤、DEAESepharoseFastFlow及SephacrylS-400柱层析纯化得多糖精品SEP。经高效液相色谱、聚丙烯酰胺凝胶电泳及纸层析鉴定其纯度。高效凝胶渗透色谱法(HPGPC)测定其分子量。体外脾淋巴细胞增殖实验测定其免疫活性。结果从海胆黄中分离纯化得到的均一多糖组分SEP,经检测其分子量为1950KD左右。脾淋巴细胞增殖实验表明SEP可显著促进脾淋巴细胞的增殖。结论从海胆黄中分离纯化得到的均一多糖组分SEP具有较强的体外免疫活性。     

灵芝孢子粉中多糖的制备工艺研究

目的：针对灵芝孢子粉多糖的制备工艺进行研究。方法：以多糖转移率为指标,比较布袋包煎和搅拌提取两种工艺;采用单因素法,考察搅拌提取中料液比和提取次数对多糖提取的影响;根据滤液的性状,比较静置取上清液、离心、布袋滤过和板框滤过等多种滤过方法,优化滤过工艺。结果：灵芝孢子粉多糖的最佳制备工艺条件为：按料液比1：30加水至提取罐中,煮至100℃,投料,保温搅拌提取2min,静置24min,取上清液,离心,滤液先通过（5μm滤袋）,再板框滤过,滤液浓缩至相对密度为1．10（60℃）,醇沉,取沉淀减压干燥,千膏得率为3．26％,多糖含量为25．1％,多糖转移率97％。结论：该工艺可行有效,具有实际生产意义。     

Fucoidan is the active component of fucus vesiculosus that promotes contraction of fibroblast-populated collagen gels

The fibroblast-populated collagen gel culture method has been evaluated as a dermal model of wound contraction and granulation in tissues during the wound healing process and as an in vitro model of dermal tissue. We previously reported that an extract of Fucus vesiculosus promoted fibroblast-populated collagen gel contraction and that the promotion of the gel contraction was due to the increased expression of integrin alpha2beta1 on the surface of the fibroblasts. In this study, we investigated the active component of the extract of this alga using extraction and fractionation techniques. Water extraction of the alga was followed by precipitation with excess ethanol and then gel filtration with the boundary molecular weight of 30,000. The high molecular weight fraction obtained from gel filtration was fractionated by ion exchange chromatography on diethylaminoethyl cellulose column to give active fractions that have more polar properties. These polar, high molecular weight fractions which contained molecules with fucose and sulfate groups showed significant gel contraction-promoting activity and integrin expression-enhancing activity, and were estimated to be the sulfated-polysaccharide fucoidan. Commercially available fucoidan showed similar activities to the above-described fraction of this alga. Although it remains necessary to precisely identify the specific active component, the above results indicate that fucoidan is the active component which promotes collagen gel contraction, and also indicate the possibility that it dose so by enhancing the integrin alpha2beta1 expression.     

Antitumor components ofAgrocybe cylindracea

To find pharmacologically active components ofAgrocybe cylindracea, its basidiocarps were extracted with water. The extracts were separated by DEAE cellulose column chromatography, Sepharose CL-4B gel filtration, and Concanavalin A-Sepharose 4B affinity chromatography. Among the obtained fractions fromA. cylindracea, fraction IN which was the neutral proteinbound-polysaccharide fraction exhibited a marked antitumor activity and it was tentatively named “Cylindan”. It showed about 70% of tumor inhibition against the solid form of sarcoma 180 when a dose of 30 mg/kg/day was intraperitoneally injected into ICR mice. When each fraction was examined by chemical analysis, Cylindan consisted of 85% polysaccharide, 3% protein and 1% hexosamine. Its polysaccharide moiety contained glucose, mannose, fucose and galactose and its protein moiety contained the comparatively large amounts of aspartic acid and glycine, and other 11 amino acids.     

Purification of an anti-ulcer polysaccharide from the leaves of Panax ginseng.

Water-soluble and alkaline-soluble crude polysaccharides which were separated from the roots or leaves of Panax ginseng C. A. Meyer, were compared for their anti-ulcer activity. Of these four polysaccharide fractions, the water-soluble crude polysaccharide fraction (GL-2) from the leaves and the alkaline-soluble crude polysaccharide fraction (GRA-2) from the roots prevented HCl/ethanol-induced ulcerogenesis in mice potently. The most potent fraction, GL-2, was further fractionated into four polysaccharide fractions by precipitation with cethyltrimethylammonium bromide, and the weakly acidic polysaccharide fraction, GL-4, showed the most potent inhibition of gastric lesion formation. The activity of GL-4 decreased after treatment with periodate or digestion with endo-polygalacturonase, indicating that the carbohydrate moiety may contribute to the expression of the activity. GL-4 was further purified by anion-exchange chromatography and gel filtration, and the most active purified polysaccharide, GL-4IIb1III was obtained. GL-4IIb1III (average relative molecular mass, 16,000 d) had the nature of a pectic polysaccharide, and was composed mainly of galactose and galacturonic acid with small proportions of rhamnose, arabinose, mannose, glucose, and glucuronic acid. GL-4IIIb1III prevented HCl/ethanol-induced ulcerogenesis in mice dose dependently.