蒺藜中水溶性多糖的研究——杂多糖H的纯化与结构初步确定

从蒺藜(Tribulus terristris L)茎和叶中提出醇溶皂甙后,再提取水溶性粗多糖,其单糖组成及其摩尔比为Ara.,Rha.,Xyl.,GalA.,Gal.,Glc.,Man.,6.0:2.1:1:3.6:3.4:7.7:2.9.粗多糖经分级与纯化得一均一级分H,其单糖组成及其摩尔比为Ara.,Rha,Xyl,GalA,Gal,Glc,1.6:2.4:0.1:3.5:1.3:1。H为酸性杂多糖,分子量约为10万。经果胶酶酶解、β-D-半乳糖苷酶酶解,高碘酸氧化、Smith降解、部分酸水解、甲基化、与GC及GC-MS分析等,表明H为α-D-GalA(1→4)与L-Rha(1→2)可能交替连接构成的主链,一些Rha.上带有分枝。     

白木通多糖的研究

用碱液从白木通(Akebia trifoliate)茎中提取所得的粗多糖ATB经DEAE-Celulose及SephadexG-200柱层析后得到一多糖纯品ATBB 2,其分子量为2.3×10⁵。糖组分分析ATBB-2中各糖残基的摩尔比为Rha∶Ara∶Xyl∶Gal∶Glc∶GalA=1.22∶1.00∶1.10∶0.85∶0.24∶0.82。经甲基化,高碘酸氧化,Smith降解,部分酸水解,¹H和¹³CNMR谱的分析揭示ATBB-2是一个结构复杂的杂多糖。     

柱前衍生化气相色谱法测定当归多糖的单糖组成

采用三氟乙酸水解当归多糖。糖腈乙酰衍生化水解单糖产物，用气相色谱法测定当归粗多糖CAP及其组分APF1、APF2和APF3的单糖组成。结果表明，当归粗多糖CAP及其组分APF1、APF2和APF3的单糖组成摩尔比各不相同。CAP由鼠李糖(Rha)、阿拉伯糖(Ara)、甘露糖(Man)、葡萄糖(Glc)、半乳糖(Gal)组成，其摩尔比为1．00：2．72：0．72：4．00：2．32；APF1由Rha．Ara、Glc、Gal组成。其摩尔比为1．00：227：7．80：269；APF2由Rha、Ara、Man、Glc、Gal组成，其摩尔比为1．00：529：3．66：9．11：5．17；APF3由Rha．Ara、Man、Glc、Gal组成。其摩尔比为1．00：454：298：11.09：7.45。     

柱前衍生化HPLC分析蒲公英多糖的单糖组成

目的 合理优化衍生方法测定蒲公英多糖中单糖的组成及摩尔比。方法 采用1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生,用反相高效液相色谱法进行测定。色谱条件为：ZORBAX-C₁₈柱;流动相：0.1 mol·L^-1的磷酸盐缓冲液(pH 6.7)-乙腈(83∶17);流速：1 mL·min^-1;检测波长：245 nm;进样量：20 μL;柱温：室温。结果 蒲公英多糖由D-鼠李糖、葡萄糖、D-半乳糖、D-木糖和D-阿拉伯糖组成,以不同方法提取的各单糖含量的摩尔比为：水提取法,Rha∶Glc∶Gal∶ Ara=0.088∶0.500∶0.190∶0.340;超声提取法,Rha∶Glc∶Gal∶Ara=0.050∶0.350∶0.080∶0.320;酶提取法,Rha∶Glc∶ Gal∶Ara=0.270∶0.630∶0.330∶0.460;超声协提取同酶法,Rha∶Glc∶Gal∶Ara =0.078∶0.550∶0.130∶0.170。结论 改进后的衍生化方法和等度洗脱的色谱方法具有操作简单、快速、重复性好等特点,可用于蒲公英多糖中单糖组成和摩尔比的测定。     

骆驼刺茎枝粗多糖的理化性质及抗氧化活性研究

摘 要 目的：提取分离骆驼刺茎枝粗多糖,测定其理化性质及抗氧化活性。方法: 采用水提醇沉法提取骆驼刺茎枝粗多糖,硫酸苯酚法测定总糖含量,考马斯亮蓝法测定蛋白含量,咔唑硫酸法测定糖醛酸含量,GC法测定单糖组成及相对摩尔比;通过测定骆驼刺茎枝多糖的还原力,及其对1,1-二苯基-2-三硝基苯肼（DPPH）自由基和羟自由基的清除力,评价骆驼刺茎枝多糖的体外抗氧化活性。结果:水提醇沉制备的骆驼刺茎枝粗多糖总糖含量为73.2%,其中糖醛酸占粗多糖总糖含量的27.2%;蛋白质含量为17.6%;骆驼刺茎枝多糖由Rha、Ara、Xyl、Man、Glc、Gal、GlcA和GalA 8种单糖组成,相对摩尔比为1.05∶1.00∶1.25∶0.52∶3.05∶1.31∶0.47∶4.78;骆驼刺茎枝多糖的还原力、对DPPH与羟基自由基的清除率随多糖浓度的增大而增强。结论:初步提取分离了骆驼刺茎枝粗多糖,研究了其理化性质和体外抗氧化活性,为骆驼刺的开发利用提供研究基础。     

柱前衍生化HPLC分析蒲公英多糖的单糖组成

目的合理优化衍生方法测定蒲公英多糖中单糖的组成及摩尔比。方法采用1-苯基-3-甲基-5-吡唑啉酮（PMP）柱前衍生,用反相高效液相色谱法进行测定。色谱条件为：ZORBAX-C18柱;流动相：0.1mol·L^-1的磷酸盐缓冲液（pH6.7）-乙腈（83∶17）;流速：1mL·min^-1;检测波长：245nm;进样量：20μL;柱温：室温。结果蒲公英多糖由D-鼠李糖、葡萄糖、D-半乳糖、D-木糖和D-阿拉伯糖组成,以不同方法提取的各单糖含量的摩尔比为：水提取法,Rha∶Glc∶Gal∶Ara=0.088∶0.500∶0.190∶0.340;超声提取法,Rha∶Glc∶Gal∶Ara=0.050∶0.350∶0.080∶0.320;酶提取法,Rha∶Glc∶Gal∶Ara=0.270∶0.630∶0.330∶0.460;超声协提取同酶法,Rha∶Glc∶Gal∶Ara=0.078∶0.550∶0.130∶0.170。结论改进后的衍生化方法和等度洗脱的色谱方法具有操作简单、快速、重复性好等特点,可用于蒲公英多糖中单糖组成和摩尔比的测定。     

半枝莲多糖B3-PS2分离纯化及抗补体活性研究

本文提取半枝莲多糖,分别经DEAE-cellulose、Sephacryl S-300柱层析分离纯化后得到均一的B3-PS2组分,并对该组分进行了抗补体活性研究。研究结果表明B3-PS2组分分子质量为1 100 kD,主要由Glc、Gal和Ara三种单糖组成,糖残基摩尔比为2.7∶2.7∶1.0,还含有少量的Man、Rha、Fuc和Xyl。B3-PS2抑制补体活性的经典途径CH₅₀为(0.23 ± 0.03) mg·mL⁻¹,抗补体作用靶点为补体C1r、C1s、C3、C4组分。半枝莲多糖B3-PS2组分体外抗补体活性接近于阳性对照肝素,表明活性较好,这显示了半枝莲多糖治疗补体过度激活相关疾病的潜在价值。     

毛细管气相色谱法测定果胶的单糖组成

目的使用毛细管气相色谱法测定果胶样品的单糖组成及摩尔比。方法采用酸完全水解法水解果胶,糖液乙酰化衍生后,用毛细管气相色谱分析其单糖的组成及摩尔比。结果果胶中鼠李糖(Rha)、阿拉伯糖(Ara)、木糖(Xyl)、葡萄糖(Glc)、半乳糖(Gal)和半乳糖醛酸(GalA)的摩尔比为7.31∶3.68∶1∶3.09∶21.48∶131.54。结论果胶的单糖组成以半乳糖醛酸和半乳糖为主,同时含有鼠李糖、阿拉伯糖、木糖、葡萄糖等单糖。     

离子色谱测定多糖的单糖组成的方法学研究及其应用

本文旨在建立一种采用高效阴离子色谱分离和定量分析11种常见单糖的方法。采用Dionex ICS-5000+离子色谱仪,以Carbo PACTMPA 10(2.0 mm×250 mm)阴离子交换柱为分离柱,脉冲安培检测器检测,200 mmol/L Na OH和200 mmol/L的CH3COONa梯度洗脱,建立了可同时检测11种常见单糖的离子色谱-脉冲安培法(HPAEC-PAD),可实现核糖(Rib)、岩藻糖(Fuc)、鼠李糖(Rha)、阿拉伯糖(Ara)、半乳糖(Gal)、葡萄糖(Glc)、甘露糖(Man)、木糖(Xyl)、果糖(Fruc)、半乳糖醛酸(Gal A)、葡萄糖醛酸(Glc A)的定性定量分析。该方法在1~50 mg/L范围内具有良好的线性(R2=0.99100~0.99978),精密度RSD值小于3.52%,回收率在97.33%~101.82%,重复性RSD值小于3.04%。采用该方法对牛蒡精制多糖ALP-1的单糖组成进行了测定,结果表明牛蒡多糖ALP-1由岩藻糖(Fuc)、阿拉伯糖(Ara)、葡萄糖(Glc)和果糖(Fruc)组成,摩尔比为7.69∶1.00∶34.62∶3.08。对牛蒡多糖单糖组成的成功测定说明该方法成功建立,为常见单糖的定性定量分析及多糖的单糖组成分析提供了快速可靠的方法。     

枸杞子中免疫活性成分的分离、纯化及物理化学性质的研究

从宁夏枸杞子提取得到的糖缀合物LBP经DEAE-celulose柱色谱得到5个组分,其中Lbp3,Lbp4和Lbp5分别经CM-SephadexC-50,SephadexG-100,SephadexG-50柱色谱得到均一组分LbGp3,LbGp4和LbGp5。N含量:LbGp30.83%,LbGp41.72%,LbGp59.58%。总糖含量:LbGp393.6%,LbGp485.6%,LbGp58.6%。分子量:LbGp39.25×10⁴,LbGp4 21.48×10⁴,LbGp52.37×10⁴。糖组成为:LbGp3:Ara∶Glc=1∶1,LbGp4:Ara∶Gal∶Rha∶Glc=1.5∶2.5∶0.43∶0.23,LbGp5:Rha∶Ara∶Xyl∶Gal∶Man∶Glc=0.33∶0.52∶0.42∶0.94∶0.85∶1。初步分析表明LbGp4是O-连接的糖蛋白。     

人参果中水溶性多糖的研究:杂多糖F的分离提纯与结构分析

人参根、茎、叶多糖的研究已有报道,人参果胶是人参根多糖中的有效成分,人参果多糖的研究尚未见报道。采用吉林省集安制药厂提取人参果皂甙后的残渣为原料,提取水溶性人参果多糖。人参果渣用热水提取,醇析后获褐色粗多糖。经纸层析、气相色谱分析,该多糖含阿拉伯糖、鼠李糖、木糖、葡萄糖、半乳糖及半乳糖醛酸六种单糖残基,含蛋白质6.25%。将粗多     

Purification and characterization of antithrombotics from Syzygium aromaticum (L.) MErr. & PERRY

Two antithrombotic polysaccharides with relatively high molecular weight (HMW) and low molecular weight (LMW) were isolated from the flower buds of Syzygium aromaticum (L.) MERR. & PERRY (clove) by anion-exchange chromatography, hydrophobic interaction column chromatography and size exclusion chromatography (LMW: EC-2B-IIIa-2, M.W. ca. 34000; HMW: EC-2C-Ia-2, M.W. ca. 103000). The LMW polysaccharide was mainly composed of Rha, Gal, GalA and Ara (molar %: 24.1, 18.9, 18.0 and 17.9, respectively) with 10.8% of sulfate and 18.2% of protein. The HMW fraction consisted of Ara, Gal, Glc and Rha (molar %: 26.0, 23.7, 17.5 and 12.4, respectively) with 15.4% of sulfate and 8.0% of protein. Both polysaccharides had the backbone of type I rhamnogalacturonan and the side chain of arabinan. Also, most of the sulfates were attached at the position 6 of 3-linked galactosyl residues. Compared to the antithrombotic activity of the HMW fraction (plasma clotting time of 145 s in APTT assay), the LMW fraction displayed a slightly low activity (90 s). However, animal studies indicated that crude LMW polysaccharide did not show acute toxicity, while the acute LD50 of the HMW fraction was approximately 2-fold lower than that of heparin.     

Isolation and characterization of an anti-complementary polysaccharide D3-S1 from the roots of Bupleurum smithii

The preliminary data from hemolytic assays indicated that the hot-water extract of the roots of Bupleurum smithii had anti-complementary activity. Further bioactivity-guided fractionation led to the isolation of D3-S1, a homogeneous form of acidic polysaccharide. D3-S1 was a branched polysaccharide with average molecular weight about 2,000,000 Da, composed of Ara, Gal and GalA in the ratio of 2.6:1.0:1.2, along with trace of Rha, Glc, Xyl and Man. Methylation analysis and NMR identified the linkages of the residues of D3-S1. Functional analysis showed that D3-S1 inhibited complement activation on both the classic and alternative pathways with CH(50) value of 0.34+/-0.02 mg/ml and AP(50) value of 0.081+/-0.003 mg/ml, respectively. Preliminary mechanism studies by using complement component depleted-sera indicated that D3-S1 selectively interacts with C1s, C3 and C4, but not C1q, C1r, C2, C5 and C9. The results suggested that D3-S1 could be of potential benefits in treatment of the complement-associated diseases.     

A polysaccharide from the stems of Rubus amabilis Focke and its immunological enhancement activity

A water-soluble polysaccharide (named RAP) was newly isolated from the stems of Rubus amabilis. Structural confirmation of the polysaccharide was provided by hydrolysis, periodate oxidation, Smith degradation, and methylation analysis, combined with nuclear magnetic resonance (NMR), capillary electrophoresis (CE), infrared spectroscopy (IR), and gas chromatography-mass spectra (GC-MS). In vitro immunological enhancement activity was characterized using the proliferative activity of spleen lymphocytes and phagocytic activity of peritoneal macrophages in mice. The polysaccharide was mainly composed of xylose, arabinose, glucose, rhamnose, galactose, mannose, glucuronic acid, and galactocuronic acid in the molar ratio of 1.0:6.9:0.8:1.1:6.9:0.3:0.5:3.3, with the average molecular weight of 26.2 kDa. The linkage types of netural monosaccharides were as follows: the arabinose was →2) Ara (1→ and galactose were Gal (1→, →3) Gal (1→, →3,6) Gal (1→, →2,3,6) Gal (1→ and →2,3,6) Gal_f (1→. Xyl (1→, →6) Glc (1→, →2) Glc (1→, →3) Rha (1→, Rha (1→ and Man (1→ were also found in the structure. RAP-B-2 could improve the proliferative activity of spleen T cells and B cells and boost phagocytic activity of peritoneal macrophages at the concentration of 50 μg/ml (p < 0.05, p < 0.01).     

Isolation and characterization of an anti-complementary protein-bound polysaccharide from the stem barks of Eucommia ulmoides

EWDS-1, a homogeneous protein-bound polysaccharide, was isolated as an anti-complementary agent from the stem barks of Eucommia ulmoides. EWDS-1 was identified as a branched proteoglycan with average molecular weight about 2,000,000 Da, composed of Gal, Glc and Ara in the ratio of 2.1:1.0:0.9, along with trace of Rha, Xyl, Man, as well as 3.95% of protein. The linkages of the residues of EWDS-1 were deduced by methylation analysis and NMR technique. Bioassay showed that EWDS-1 inhibited complement activation on both the classic and alternative pathways with CH(50) and AP(50) values of 203+/-20 microg/ml and 45+/-8 microg/ml, respectively. Preliminary mechanism studies by using complement component depleted-sera indicated that EWDS-1 inhibits activation of complement system by interacting with C1q, C1r, C1s, C2, C3, C4, C5 and C9. The results suggested that EWDS-1 could be of promising benefits in treatment of the complement associated diseases.     

仙鹤草多糖的分离纯化及理化性质研究

目的从中药仙鹤草中提取仙鹤草多糖,并对其理化性质进行研究。方法利用水提醇沉法得粗多糖,并依次经纤维素DE-52柱和Superdex-200凝胶柱色谱进行纯化。采用光谱法和色谱法对其总糖含量、单糖组成、相对分子质量等理化性质进行研究。结果分离纯化的仙鹤草多糖HAPⅢ总糖含量、蛋白含量、糖醛酸含量分别为81%,3.4%,45%,由半乳糖、葡萄糖、鼠李糖、阿拉伯糖、甘露糖、岩藻糖、木糖组成,摩尔比为5.65∶2.67∶2.61∶1.86∶1.12∶1.09∶1。其平均相对分子质量为1.90×105。结论仙鹤草多糖HAPⅢ是一水溶性酸性杂多糖。     

人工培养蛹虫草多糖的分离纯化及其结构的初步研究

目的人工培养蛹虫草多糖类成分的提取与纯化。方法采用SephadexG 10 0柱色谱法进行了分离纯化 ;通过理化常数测定、化学和光谱分析等方法研究了其化学结构特征。结果分离得到CPS 1、CPS 2和CPS 33种多糖 ,其单糖组成分别为 :CPS 1:鼠李糖 木糖 甘露糖 葡萄糖 半乳糖 (1∶6 .4 3∶2 5 .6∶16 .0∶13.8) ;CPS 2 :鼠李糖 葡萄糖 半乳糖 (1∶4 .4 6∶2 .4 3) ;CPS 3为只含葡萄糖的均一多糖。 3种多糖的分子量分别为 2 30 0 0、12 90 0和 5 0 0 0。CPS 1主要含有β糖苷键 ,CPS 2和CPS 3主要含有α糖苷键。 结论CPS 1为一种含有 5种单糖 ,糖苷键为 β构型的多糖 ;CPS 2是一种含有 3种单糖 ,糖苷键为α构型的多糖 ;CPS 3是一种仅含葡萄糖、糖苷键为α构型的多糖