凝胶排阻色谱-多角度激光散射法测定破伤风类毒素相对分子质量分布

为更好地表征精制破伤风类毒素(精制TT)相对分子质量分布的情况,利用凝胶排阻色谱与多角度激光散射仪联用法(SEC-MALS法)对精制破伤风类毒素分子质量及其分布进行了初步探索.通过优化实验参数,确定了最佳实验条件:流动相为0.2 mol/L磷酸盐缓冲液-1％异丙醇(pH =7.0)(PBS),样品浓度为2.0 mg/m...     

凝胶色谱与多角激光光散射联用研究聚氧化乙烯的扩展效应和链构象

采用凝胶色谱与多角激光光散射联用的方法,测定了一系列不同分子量的聚乙二醇(PEG)和聚氧化乙烯(PEO)在色谱柱中的扩展效应.扩展因子随PEG/PEO分子量的增加而增大,经扩展效应改正后得到了样品的准确分子量和分子量分布.同时建立了PEO的Z均回转半径Rgz与重均分子量Mw之间的单分散标度关系:Rgz=0.0272 Mw0.56,结果表明,长链PEO在水溶液中由于排除体积效应采取溶胀的无规线团构象.     

凝胶色谱-光散射联用法对壳聚糖及其产品的分子质量及分布的测量

采用国家基准物质氯化钠和甲苯校准了示差折光仪和增强光系统的仪器常数,用超纯水重量法校准了激光光散射仪到示差检测器之间的延时体积,用葡聚糖标准物质对各角度的光强进行归一化处理,研究了凝胶色谱-激光光散射联用方法中设备的校准和溯源。上述校准途径使仪器溯源到国际SI单位,从而确保了测量结果准确可靠。采用激光光散射仪测得壳聚糖及其产品术后防粘连隔离膜的光折射增量dn/dc值分别为0.118、0.092 mL/g,凝胶色谱-激光光散射联用方法测得壳聚糖及其产品术后防粘连隔离膜的重均分子质量分别为8.897×104、1.168×105g/mol;分子质量分布Mw/Mn分别为1.148±0.026和3.132±0.377。     

体积排阻色谱和激光光散射对大豆蛋白热诱导聚集体的研究

采用体积排阻色谱(SEC-HPLC)和激光光散射(LLS)研究了不同浓度和离子强度下大豆蛋白热诱导聚集体的分子量分布和粒径分布。在离子强度为0时,SEC-HPLC的结果表明,热处理后的蛋白溶液主要由3部分组成,即聚集体、中间体和未聚集部分。聚集体部分随着浓度增加而逐渐增加;LLS的结果表明:体系有不均一的粒径分布,且浓度增加时体系的平均粒径增加。上述样品在较高离子强度下加热时,SEC-HPLC和LLS的结果都证明溶液中的中间体部分逐渐消失。因此,控制浓度和离子强度可以制备不同性质的大豆蛋白聚集体。     

Determination of Morphological Properties of Swellable Solids by Size Exclusion Chromatography

Many technically interesting porous solids, e.g. ion exchangers or adsorbents for catalysis, are swellable polymers, i.e. the pore structure depends on the solvent medium. A method based on exclusion chromatography, permits determination of the pore size and pore size distribution in the swollen state.—Size exclusion chromatography, also referred to as gel permeation, gel filtration, or molecular sieve chromatography, is a widely employed method for the separation of dissolved substances—mostly polymer mixtures—according to their molecular size. Porous solids are used as stationary phase. Conversely, pore sizes and other structural data can be determined by exclusion chromatography. This application requires a series of standards (polymer samples) of known molecular weight. As a simple and rapid method, it has already proven valuable for such determinations in the case of rigid solids; in the case of swellable solids, this constitutes the sole method by which the pore structure can be characterized: classical methods require dry samples.     

体积排阻色谱法测定胸腺肽分子量校准曲线条件的优化

在体积排阻色谱(SEC)法测定胸腺肽分子量校准曲线过程中,流动相中乙腈的比例对核糖核酸酶A、人胰岛素、胸腺肽α1和生长激素释放抑制因子4种蛋白的保留时间有重要影响,进而影响校准曲线的线性关系。当乙腈比例为75%时,胸腺肽分子量校准曲线线性最好,此时分子量校准方程为y=-3.138 6x+21.724,线性相关系数r2=0.988 5。4种蛋白的理论塔板数在45 783～63 345之间,拖尾因子在0.96～1.18之间,分离度在3.52～8.82之间。SEC法测定胸腺肽分子量校准曲线的液相色谱条件对4种蛋白的分离效果优异,分子量校准曲线线性良好,可用于胸腺肽制剂中高分子量物质的检测。     

Characterization of River Natural Organic Matter by High-Performance Size Exclusion Chromatography

The molecular weight of natural organic matter from the Yeongsan and Seomjin Rivers and the discharge from the wastewater treatment plants were analyzed for seasonal characteristics by high-performance size exclusion chromatography. The distribution of molecular weights demonstrated relationships with microbial activity and seasonal variations with temperature. Dissolved organic carbon and total organic carbon for the Seomjin River were from 1.6 to 3.1 mg L^?1; for the Yeongsan River system, these parameters were between 2.0 and 8.0 mg L^?1. There were no significant differences between upstream and downstream organic matter. The biochemical oxygen demand (0.8–2.5 mg L^?1) for the Seomjin River was less than the chemical oxygen demand (2.9–5.0 mg L^?1). Similarly, the biochemical oxygen demand (0.9–4.6 mg L^?1) for the Yeongsan River was lower than the chemical oxygen demand (3.8–10.6 mg L^?1). The molecular weight of aromatic compounds increased as the water temperature decreased in October, whereas the molecular weights of proteins decreased as the water temperature increased. These results suggest that as microbial activity increased, natural organic matter with a smaller molecular weight may be employed as an energy source by microorganisms. As a result, seasonal variation of the water temperature may influence the distribution of organic matter in the rivers based on molecular weight and microbial activity such that the smaller natural organic matter was more readily degraded by microorganisms.     

Determination of Polysaccharides in Undaria pinnatifida by Ionic Liquid-Modified Silica Gel Size Exclusion Chromatography

Ionic liquid-modified silica particles with a large pore size were synthesized and used as the stationary phase for size exclusion chromatography coupled with a refractive index detector for the determination of polysaccharides, such as fucoidan, alginic acid, and laminarin from Undaria pinnatifida (seaweed). The molecular weight of polysaccharide was determined by a dextran standard curve (5–1100?kDa). The ionic liquid-modified silica column exhibited excellent size exclusion properties in separating laminarin from other components. The 1-butyl-3-methyl-imidazolium bis-(trifluoromethylsulfonyl)-amino silica column has superior resolution in laminarin separation than the other columns because the amino-group in ionic liquid provide π–π interactions due to aromaticity of the ring structure, which enhances the effect of the hydroxyl group in the target compound separation. The concentrations of polysaccharides were calculated using a standard linear equation to be 0.332–0.484?mg/g of fucoidan, 0.207–0.301?mg/g of alginic acid, and 0.154–0.297?mg/g of laminarin.     

非抑制型电导检测离子排斥色谱法分析有机酸

本文以对甲苯磺，酸加硼酸作为淋洗液，采用非抑制型电导检测，研究了离子排斥色谱法分离和检测苹果酸，抗坏血酸，乳酸，琥珀酸，甲酸，乙酸，丙酸等有机酸的色谱条件，并进行了实际样品的分析。     

体积排阻色谱法研究变性溶菌酶分子复性过程中集聚体的形成

在体积排阻色谱柱上研究了还原剂存在时脲和盐酸胍变性的3种溶菌酶溶液的复性和分离过程。当变性液中原始溶菌酶浓度大于10 g/L时,变性溶菌酶在体积排阻色谱柱上除了复性为与未变性溶菌酶出峰时间相同的复性态溶菌酶分子外,还形成了溶菌酶折叠中间体的二分子集聚体。这个结果得到了用稀释法复性时溶菌酶的蛋白电泳检测结果的支持。与稀释法复性相比较,用体积排阻色谱法复性时所形成的折叠中间体二分子集聚体的量要远远低于用稀释法所形成的集聚体的量。     

凝胶排阻色谱法测定中空纤维超滤膜组件对聚乙二醇的截留率

采用凝胶排阻色谱法测定中空纤维超滤膜组件对聚乙二醇20000(PEG20000)溶液的截留率,选择Agilent PL aquagel-OH MIEXD–H8μm色谱柱,流动相为0.1 mol/L Na NO3溶液(含0.02%Na N3),流量为1 m L/min,RID检测器,柱温箱和检测器温度均为30℃。当进样体积为50μL时,方法的检出限为18.8 mg/L,定量限为49.0 mg/L。PEG20000溶液的质量浓度在60~1 000 mg/L范围内与色谱峰高呈良好的线性,相关系数r2=1.000。该方法对实际样品测定结果的相对标准差为1.31%(n=6)。将该方法的测定结果与紫外–可见分光光度计法的测量结果进行F检验和t检验,结果表明两种方法测量结果无显著性差异。采用该方法可以同时对PEG20000样品的相对分子质量及其分布进行测定,从而对选用的截留标准物质进行质量控制。     

一种快速测定牦牛皮胶和牛皮胶中18种氨基酸的PITC柱前衍生HPLC法

建立一种有效测定牦牛皮胶与牛皮明胶中18种氨基酸含量的异硫氰酸苯酯(PITC)柱前衍生-高效液相色谱法。对PITC衍生方法进行了优化,方法无需干燥和有机溶剂萃取过量PITC的步骤。牦牛皮使用胃蛋白酶在37℃下酶解72 h,酶解物冷冻干燥保存。样品用6.0 mol/L盐酸,110℃水解后,以PITC为衍生试剂进行衍生处理。采用Shiseido Capcell Pak C18(4.6 mm×250 mm,5μm)色谱柱,流动相为0.14 mol/L乙酸钠-0.5 m L/L三乙胺水溶液(p H 6.08)和60%乙腈,梯度洗脱,流速为1 m L/min,检测波长为254 nm。18种氨基酸在0.2~80 mg/L浓度范围内与峰面积间的线性关系良好(r2≥0.999 2),检出限为0.01~0.39 mg/kg,平均回收率为78.7%~121.0%,相对标准偏差(RSD)为0.05%~12.0%。结果表明,该方法衍生化步骤简单、灵敏度高、前处理时间短,可用于牛科动物皮胶中多种氨基酸成分和含量的测定。