红花均一多糖的结构表征及抑制HepG2增殖活性

对红花多糖进行分离提纯和结构表征,探讨了红花均一多糖对HepG2肝癌细胞增殖的抑制作用.采用水提醇沉法提取红花粗多糖;通过阴离子交换色谱柱和分子筛凝胶色谱柱对红花粗多糖进行分离纯化;采用高效凝胶渗透色谱法(HPGPC)对红花多糖的均一性以及分子量分布进行测定;采用单糖组成、甲基化及核磁共振波谱等方法对其进行结构解析.通过体外细胞实验,研究了红花多糖对于肿瘤细胞的作用.从红花中分离得到一种均一多糖(CTPS-1A),通过HPGPC计算得出其分子量为53800. CTPS-1A的单糖组成为n(阿拉伯糖)∶n(半乳糖)=59.2∶40.8.利用甲基化分析其连接方式为T-Galp, 1,3-Galp, 1,6-Galp, 1,3,6-Galp, T-Araf,1,5-Araf和1,3,5-Araf.研究结果表明, CTPS-1A为阿拉伯半乳聚糖(AG),其主链由-β-Galp-(6→1)-β-Galp-(6→组成,并在1,6-β-Galp的3位连有T-β-Galp, 1,3-β-Galp, T-α-Araf, 1,5-α-Araf, 1,3及5-α-Araf等分支.体外培养HepG2细胞实验结...     

柱前衍生化-高效液相色谱法分析缅甸刺梧桐胶多糖的单糖组成

利用1-苯基-3-甲基-5-吡唑啉酮柱前衍生化-高效液相色谱法,优化10种常见单糖的色谱分离条件,并用于不同等级的自制刺梧桐胶多糖的单糖组成分析。对自制刺梧桐胶多糖的单糖组成分析结果表明,自制刺梧桐胶多糖由葡萄糖醛酸(GlcUA)、鼠李糖(Rha)、半乳糖醛酸(GalUA)、半乳糖(Gal)、阿拉伯糖(Ara)和岩藻糖(Fuc) 6种单糖组成,平均物质的量比为1∶3.30∶1.62∶2.45∶0.08∶0.03。对不同等级的刺梧桐胶多糖的分析鉴定结果表明,不同等级的刺梧桐胶多糖的单糖组成均含Rha、GalUA、GlcUA、Gal 4种单糖组分,其中Ara在缅甸A、B级中没有检测出来,Fuc在缅甸D级中没有检测出来。方法学结果显示:10种单糖在各自线性范围内线性关系良好,相关系数r≥0.9992,6种单糖加标回收率在96.80%～101.26%之间,相对标准偏差小于3%。该方法简单、准确灵敏,可用于刺梧桐胶多糖的单糖组成分析。     

山茱萸酸性多糖FCP5-A的分离纯化与结构表征

从山茱萸中提取出水溶性粗多糖, 经柱色谱分离纯化得到一种酸性多糖组分FCP5-A. 采用高效凝胶渗透色谱法(HPGPC)测定其为均一性多糖, 平均分子量为8.7×104. 经IR、GC、部分酸水解、13C NMR及甲基化分析等方法对该多糖的化学结构进行了表征. 结果表明, 该多糖由鼠李糖、阿拉伯糖、半乳糖及半乳糖醛酸组成, 其摩尔比为1∶5.7∶0.6∶1.2. FCP5-A为多分支结构, 由-2)Rha(1-及-4)GalA(1-构成主链, 在鼠李糖的4位存在分支; 支链主要由高度分支的阿拉伯糖构成, 此外还存在-3)Gal(1-; 末端残基为Ara(1-及Gal(1-. 结果提示, FCP5-A为一种新的山茱萸酸性分支多糖.     

高效凝胶渗透色谱及高效液相色谱和电喷雾-离子阱质谱法联用测定构成芦荟多糖的单糖

利用高效凝胶渗透色谱(HPGPC)测定芦荟浓缩液和芦荟粉中多糖的纯度和相对分子质量,使用1-苯基-3-甲基-5-吡唑啉酮(PMP)作为柱前衍生试剂分别对样品和其水解产物进行衍生,结合高效液相色谱(HPLC)分离和电喷雾-离子阱质谱(ESI-MS)检测分析其单糖组成.结果表明:组成芦荟中多糖的单糖主要是甘露糖和葡萄糖.     

独脚金水溶性多糖的结构表征及其清除自由基活性

独脚金粗多糖(Strigaasiatica(L.)O.Ktze. polysaccharide,SKPc)经DEAE-Sephadex A-25以及Sephadex G-100柱层析分离纯化后得到独脚金多糖组分(SKP~*_1)。采用高效液相色谱(HPLC)法检测SKP~*_1,表明多糖SKP~*_1极性、相对分子质量是均一的,且其相对分子质量为14852u。采用气相色谱(GC)分析法研究SKP~*_1的单糖组成为葡萄糖(Glu)、阿拉伯糖(Ara)、半乳糖(Gal)、木糖(Xyl)、甘露糖(Man)、鼠李糖(Rha),相对摩尔比是2.5∶2.0∶0.85∶0.4∶0.5∶1.0;可能主要以β型糖苷键连接。采用化学分析和仪器分析结合进行SKP~*_1结构分析,表明SKP~*_1结构是Ara以β-(1→3)糖苷键连接构成主链之一,2,4或2,5位有支链;Gal以β-(1→6)糖苷键连接构成SKP~*_1的主链之一,且在3位上有分枝。支链由β-(1→4或1→5)Xyl、β-(1→4,1→6,1→2)Glc构成。Rha、Ara、Man、Gal、Glc都有一部分构成糖分子的末端残基。体外清除羟基实验结果表明,SKP~*_1有较好地清除羟基自由基的活性。     

沙蒿籽水溶性胶多糖ASPI-A的结构与免疫活性的初步研究

从沙蒿籽中提取出水溶性胶多糖,经柱色谱分离纯化得到一种中性多糖组分ASPI-A.采用高效凝胶渗透色谱法(HPGPC)测定其为均一性多糖,平均分子量为5.42×104Da.经IR,GC部分酸水解、甲基化分析等方法对该多糖的化学结构进行了表征.结果表明,该多糖由阿拉伯糖、甘露糖、葡萄糖及半乳糖组成,其物质的量的比为1∶2.8∶4.9∶1.9.ASPI-A为多分支结构,以(1→4)-β-Glc构成主链,部分葡萄糖C6存在分支,由甘露糖以-4)Man(1-连接在葡萄糖C6位,Glc(1-和Gal(1-连接在甘露糖C4位构成.免疫活性实验结果表明,ASPI-A在10～50μg·mL-1浓度范围内对ConA诱导的小鼠T淋巴细胞增殖反应具有促进作用.     

高效液相色谱串联质谱法测定牛奶中的高氯酸盐

建立了高效液相色谱-串联质谱测定牛奶中高氯酸盐的方法.样品经1%乙酸-乙腈(体积比1:4)混合溶液提取,于6 000 r/min离心20 min后,经0.2μ m的尼龙滤膜、On-GuardⅡRP柱、On-GuardⅡAg柱和On-GuardⅡBa柱净化,最大反相性能色谱柱C12(Synergi 4u MAX-RP 8...     

凝胶过滤色谱法测定泥鳅多糖的组成及分子量

从泥鳅粘液中分离提取到中性的粗多糖 ,采用凝胶过滤色谱柱在高效液相色谱仪上进行分离和测定 ,结果表明 ,该多糖由一个高聚糖 (MAP ,19 1% )和一个寡糖 (MAO ,80 9% )组成。用标准的Pullulan(P 112 0 0 0 ,P 4 730 0 ,P 2 2 80 0 ,P 5 90 0 ,P 2 70 0 )作标准曲线 ,测得其重均分子量分别为 130 2 5 2 (约 72 3个糖基 )和 15 39(约 9个糖基 )。粗多糖经SephadexG 10 0凝胶柱 (1cm× 5 0cm)过滤 ,分离得到纯化的MAP和MAO。     

佛手多糖的化学组成及体外抗氧化活性研究

佛手经热水提取和乙醇沉淀,得佛手多糖粗品(BP).BP经Servag法除蛋白,用DEAE离子交换柱梯度洗脱分离得到BP1和BP2两个组分,经凝胶色谱和SDS-聚丙烯酰胺凝胶电泳检验为均一组分.用凝胶渗透色谱法测得BP1和BP2的重均分子量分别为17773和65589.薄层色谱和高效液相色谱分析表明,BP1和BP2均由鼠李糖、甘露糖、葡萄糖、半乳糖和木糖组成.采用化学发光法在多种化学模拟体系中研究了佛手多糖清除活性氧的作用,观察了佛手多糖对HO^.导致DNA链损伤的抑制作用.结果表明,佛手多糖能有效地清除O₂^-.和HO^.等活性氧,对DNA链具有良好的保护作用.     

花生致敏糖蛋白Ara h1糖链决定簇的质谱分析

以花生种子总蛋白及其主要致敏糖蛋白Ara h1为研究对象,采用"一釜法"对蛋白上的糖链进行释放并同时进行衍生化标记,通过C18固相萃取柱纯化,以电喷雾质谱(ESI-MS)、多级串联质谱(MSn)和亲水性液相色谱-质谱联用(HILIC-MS)进行结构解析和定量分析.结果表明,蛋白Ara h1共有10条N-糖链,其中7条为高甘露糖型,2条为木糖修饰,另外1条为与过敏原相关的核心α1,3-Fuc修饰N-糖链,其含量约占总糖链的12.45%.     

白芷多糖的分析

研究了白芷多糖的结构和单糖组成。采用紫外光谱和红外光谱法对白芷多糖进行定性分析。结果表明,白芷多糖具有多糖特征性的紫外和红外吸收峰,其分子中存在酰胺结构;凝胶渗透色谱法测定结果表明,白芷多糖的峰值分子量为88538;采用气相色谱法测定白芷多糖中单糖的种类和构成比例,结果表明,白芷多糖由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖和半乳糖等7种单糖组成,并计算出其中6种单糖的摩尔组成比例。     

枸杞功能性成分的提取及其组成的液相色谱分析

对枸杞子中的水溶性多糖成分进行了分离提取及组成分析。采用水提、醇沉等方法提取枸杞子中的多糖成分,用Sevage法除去蛋白质后,用凝胶色谱法进行多糖的纯化,制备3-甲基-1-苯基-2-吡唑啉酮（PMP）衍生物进行液相色谱分析。结果表明,枸杞多糖由鼠李糖、阿拉伯糖、木糖、葡萄糖、半乳糖5种单糖组成,其含量分别为5．28％,17．87％,38．29％,17．82％,20．74％。     

Structural Analysis of Water-soluble Polysaccharide PIP1 Extracted from the Cultured Mycelium of Phellinus igniarius

IntroductionPhellinus igniariusbelongs to the phylum Basidio-mycota and is widely distributed in China, Japan, Aus-tralia, Philippines, and North America. The myceliumofP igniariusis perennial and grows on the stock oflatifoliate trees, such as robur, bir…     

Purification of polysaccharide from artificially cultivated Anoectochilus roxburghii (wall.) Lindl. by high‐speed counter current chromatography and its antitumor activity

To establish a systematic method for the extraction, purification, characterization and antitumor activity study of polysaccharide from artificially cultivated Anoectochilus roxburghii (wall.) Lindl. (AC‐ARPS). High‐speed counter current chromatography with two‐phase aqueous systems was successfully applied to purify AC‐ARPS after one‐step separation. The purity of the AC‐ARPS obtained by phenol/sulfuric acid method was 95.01%. The chemical structures of AC‐ARPS were identified by a series of analytical methods including high‐performance liquid chromatography and liquid chromatography with mass spectrometry. High‐performance liquid chromatography and liquid chromatography with mass spectrometry indicated that AC‐ARPS was mainly composed of mannose, ribose, glucose, galactose and arabinose with a molar ratio of 1.00:8.47:47.30:1.17:1.19. AC‐ARPS is a homogeneous polysaccharide with a molecular weight of 25 681 Da. The antitumor effect of AC‐ARPS was evaluated on lung cancer A549, osteosarcoma 143B, rat adrenal pheochromocytoma PC 12, breast cancer MCF‐7, acute leukemia HL 60, chronic leukemia K562, colon cancer SW620, esophageal cancer OE 19, liver cancer HepG2, and neuroglioma U251 cells in vitro. AC‐ARPS showed the best inhibitory effect on OE 19 cells, and the IC₅₀ value was 5.67 ± 0.831 μmol/L. Fluorescence analysis and flow cytometry results showed that AC‐ARPS induced apoptosis and G2/M phase arrest in OE 19 cells.     

Rapid separation of polysaccharides using a novel spiral coil column by high‐speed countercurrent chromatography

The separation of polysaccharides is time consuming. We developed and optimized a type‐J counter‐current chromatography system with a novel tri‐rotor spiral coil column for the rapid separation of polysaccharides. The optimal composition of an aqueous PEG1000/K₂HPO₄/KH₂PO₄ system was found to be 14:16:14 w/w/w where the lower phase was the mobile phase. Optimal performance was achieved at a column rotational speed, temperature, and flow rate of 1200 rpm, 45°C, and 3.0 mL/min, respectively. The mobile phase was pumped from the inner terminal in a ‘‘head‐to‐tail’’ elution mode. Polysaccharide LCP‐1 (10.7 mg) was successfully obtained in high purity in one step from 50.0 mg of a crude polysaccharide extracted from the lychee fruit (Litchi chinensis) within 100 min. LCP‐1 possess a number‐average molecular weight and weight‐average molecular weight of 1.05 × 10⁵ and 1.59 × 10⁵ kDa, respectively. The monosaccharide composition consists of the molar ratio of glucose, galactose, and arabinose of 1.3:3.5:1.     

Isolation and characterization of methoxylated flavones in the flowers of Primula veris by liquid chromatography and mass spectrometry

Characterization of six flavones, which were named substances G1, G2, G3, G4, G5 and G6 according to their R_F values in normal-phase thin-layer chromatography, is reported. The pure flavones were purified after maceration with methanol by normal-phase solid-phase extraction, normal-phase medium-pressure liquid chromatography, normal-phase preparative thin-layer chromatography and preparative reversed-phase high-performance liquid chromatography (RP-HPLC). The collected fractions of several isolation steps were analyzed by normal-phase (NP) and RP-HPLC. Detection and identification of the substances G was accomplished by UV detection at 213–216 nm, diode array UV detection, or fluorescence detection (λ_ex=330 nm; λ_em=440 nm). The molecular mass, the elementary composition, and the structure of the six components was determined by electron-impact high-resolution mass spectrometry (EI-HRMS). Substance G4 was identified as 3′,4′,5′-trimethoxyflavone. The substances G1–G6 were shown to be mono-, di- tri- and pentamethoxyflavones. HPLC–electrospray ionization tandem mass spectrometry (ESI-MS–MS) of the flavones was carried out employing a 150×2 mm I.D. column packed with a 3 μm/100 Å octadecylsilica stationary phase and a mobile phase comprising 1.0% acetic acid in water–acetonitrile (50:50). Comparative RP-HPLC–ESI-MS of the raw methanol extract and the isolated substances G1–G6 proved that the isolated compounds were pure and were not artifacts. Finally, RP-HPLC–ESI-MS–MS was used to identify substances G1–G6 in phytopharmaceutical drugs.     

Molecular weight and monosaccharide composition of Astragalus polysaccharides

Two polysaccharides (APS-I and APS-II) were isolated from the water extract of Radix Astragali and purified through ethanol precipitation, deproteination and by ion-exchange and gel-filtration chromatography. Their molecular weight was determined using high performance liquid chromatography and gel permeation chromatography (HPLC-GPC) and their monosaccharide composition was analyzed by TLC and HPLC methods, using a refractive index detector (RID) and an NH(2) column. It was shown that APS-I consisted of arabinose and glucose and APS-II consisted of rhamnose, arabinose and glucose, in a molar ratio of 1:3.45 and 1:6.25:17.86, respectively. The molecular weights (Mw) of APS-I and APS-II were 1,699,100 Da and 1,197,600 Da, respectively.     

柘树根多糖的分离纯化及结构表征

以柘树[Cudrania tricuspidata(Carr.) Bur.]的根为材料, 经热水抽提、木瓜蛋白酶-Sevag法除蛋白、乙醇沉淀和DEAE-Sephadex A-50凝胶柱层析分离纯化, 得到一种水溶性的柘树根多糖(CPS-0). 采用HPLC、糖基组成分析、甲基化分析、GC、GC-MS、NMR(1H NMR, 13C NMR及HMQC)、元素分析、UV和IR等技术对CPS-0的纯度、性质、组成和结构进行表征. 结果表明, CPS-0仅含葡萄糖, 分子量为4.6×103, 主链由1,4-连接的α-D-葡萄糖残基组成, 其侧链由末端及1,4-连接的葡萄糖残基构成, 取代于主链分支点葡萄糖的6位, 平均每10个葡萄糖残基组成的重复单元中含有1个分支.     

Determination of the compositions of polysaccharides from Chinese herbs by capillary zone electrophoresis with amperometric detection

In this paper, capillary zone electrophoresis with amperometric detection (CZE-AD) was applied to determine the compositions of hetero-polysaccharides from Chinese herbs, Angelica sinensis and flax by analyzing their hydrolyzed monosaccharides: fucose, galactose, glucose, arabinose, rhamnose and xylose. Under the selected optimum conditions, the six monosaccharides could be perfectly separated within 25 min and showed significant current responses at copper electrodes. The linear ranges of the six monosaccharides were all from 5.0 x 10(-6) to 2.0 x 10(-4) mol L(-1) and their detection limits were lower or near 1.0 x 10(-6) mol L(-1) (S/N = 3). Experiments showed that the Angelica sinensis polysaccharide was composed of fucose, galactose, glucose, arabinose, rhamnose and xylose (mole ratio 1.0:13.6:15.0:8.7:21.3:3.7), and the flax polysaccharide was composed of galactose, glucose and arabinose (mole ratio 1.0:4.98:1.1). The purity of these polysaccharides leached by the introduced leaching method was 98.3 and 97.6%, respectively. Analyzing polysaccharides by this method has some merits of speed, simple instrumentation and operation, high sensitivity and high reproducibility.