疏水作用色谱中流动相组成对溶质保留行为的影响

 首次研究了疏水作用色谱 (HIC)中芳香醇同系物在不同种类盐流动相中的保留行为。以计量置换保留模型中的参数Z分析了HIC中小分子与生物大分子保留行为的差别 ,以及不同流动相组成对两种类型溶质的洗脱范围及洗脱能力的影响。与反相色谱相似 ,芳香醇在HIC中的保留仍存在同系物规律。比较了小分子和生物大分子在不同盐溶液中的Z值变化 ,表明流动相中的盐仅改变小分子与固定相的水合程度 ,而对生物大分子 ,除改变其和固定相水合程度外 ,还会影响生物大分子与固定相接触区的分子构象     

体液直接进样固定相的研究进展

评述了生物体液直接进样主效液相色谱固定相的制备、结构性能及应用。其特点可概括为：只允许生物体液中蛋白质等大分子物质与亲水的生物相容的固定相表现接触、使其在柱中无保留地洗脱、而小分子物质可获得保留、达到直接进样分析的目的。     

人血白蛋白结合位点II的分子结合模式研究

人血白蛋白是血浆中含量最高的蛋白,是一种重要的载体蛋白,能与多种内源性和外源性物质结合。人血白蛋白主要有两个药物结合位点,位点I和位点II,其中位点II的柔性较大,对药物分子的亲和性较高。本文采用分子模拟方法,基于12种结合于位点II的小分子-人血白蛋白晶体结构,分析了相互作用能,发现12种分子的结合以疏水作用为主,静电作用为辅。进一步采用丙氨酸扫描和结合能评价,分析结合部位的关键氨基酸残基,探究结合模式的规律性,发现从位点入口到空腔内部存在静电、疏水和杂合的三层相互作用分布,共同形成了稳定的分子结合。最后采用分子对接和分子动力学模拟,预测了L-色氨酸的结合模式。研究结果有助于深入了解人血白蛋白药物位点II的小分子结合模式,为基于位点II的药物和分离配基的优化设计提供指导。     

蛋白质在疏水色谱法中的热行为

考察了一系列标准蛋白质在疏水色谱法中的热行为;研究了柱温变化对蛋白质和小分子保留值的影响。实验结果表明,每个蛋白质的保留值－温度曲线上都有一个固定的热变性转变温度。在这个温度以下,除热稳定性较高的溶菌酶外,蛋白质的保留值都随柱温的升高而线性地增加。在转变温度以上,保留值随柱温升高会迅速增大或减小。与蛋白质相反,小分子的保留值随柱温的升高会不断地减小。研究了柱温变化对酶活性的影响,并与酶的静态热失活进行了比较。发现仅在室温下疏水色谱法才具有温和的分离条件,在较高柱温下疏水固定相可以诱导蛋白质变性。     

体液直接进样高效液相色谱法测定尿中肌酐和尿酸

肌酐和尿酸是人体重要的代谢产物,二者均随尿排出.在体液中二者的含量及其比值是临床化学的重要参数.已报道的肌酐和尿酸的高效液相色谱分析方法常需要费时的样品预处理或使用复杂的柱切换设备,而且分析中还可能受其它物质的干扰,给分析工作带来麻烦.本实验室合成的聚氧乙烯交联γ.氨基两基固定相属于生物体液直接进样高效液相色谱固定相,在固定相疏水的表面键合亲水的基团,避免了体液中蛋白质变性吸附在柱中,同时利用填料的小孔径对蛋白质的体积排阻作用,使色谱柱对蛋白质不保留,而小分子物质则可以达到分离的目的.曾对该固定相考查了蛋白质回收实验,回收率达95%以上,并测定了实际样品,具有满意的柱效及柱寿命.本文利用该固定相直接进样分析了人尿中肌酐和尿酸,方法的线性范围宽,灵敏度高,准确性好,省时省力,为临床化学提供了一个有益的分析手段.     

吸附固定相开管毛细管电色谱方法的建立（英文）

 首次将管壁吸附作用作为开管毛细管电色谱固定相制备的推动力,成功地建立了称为“吸附固定相开管毛细管电色谱”的一种新方法。原理是：选择合适的条件,让荷正电的化合物在毛细管管壁上充分吸附,直接用吸附层作为固定相。目前,已有数类化合物被用作固定相物质,其中包括阳离子表面活性剂如十六烷基三甲基溴化铵（CTAB）、碱性蛋白质如溶菌酶和细胞色素C、碱性小肽如赖氨酸-酪氨酸和赖氨酸-丝氨酸-酪氨酸、以及碱性氨基酸如L-赖氨酸。CTAB吸附固定相用于分离电中性化合物,其它吸附固定相用于手性分离。     

芘基丁酸键合硅胶固定相对含π电子物质的高效液相色谱分离研究

通过芘基丁酸键合硅胶固定相对含π电子物质的分离研究, 考察了该固定相的色谱性能. 结果表明, 该固定相不仅具有疏水作用和电荷转移性能, 还具有很强的平面识别能力, 可很好地分离烷基苯、 稠环芳烃及取代硝基苯等各种含π电子物质， 且在pH 3.5～7.0 时化学稳定性能良好, 硅醇基效应得到了很好的抑制, 可有效地用于碱性化合物的分离分析.     

三元醇丙烯酰胺键合色谱固定相的制备及评价

利用巯基与乙烯基的"点击化学"反应合成了一种新型含多羟基的硅烷偶联剂,再将其与硅胶反应制得含多羟基的亲水固定相。经过元素分析表征证明多羟基官能团已成功键合到硅胶表面。采用一系列不同性质的标准物质考察了亲水色谱模式下固定相的溶质保留机理。由于固定相结构中既具有极性多羟基官能团,也有短的疏水碳链,因此固定相兼具疏水性与亲水性。将此固定相成功应用于亲水与反相色谱两种模式,并对比了两种模式下流速对柱效的影响。最后将固定相应用于烷基苯、水溶性维生素以及核苷的分离中,取得了较好的分离效果,证明了固定相良好的应用前景。     

胺型键合固定相分离碱性物质的影响因素考察

 以某些碱性物质为探测溶质 ,中性物质乙苯为参照溶质 ,详尽考察了检测温度、缓冲液中阳离子浓度、流动相的流速分别在流动相的 pH值为 3 0和 7 0时对胺型键合固定相分离碱性物质的影响。研究结果表明 ,随着温度的升高 ,碱性物质的保留时间先下降较快 ,然后趋于平缓 ,而柱效和峰形分别得到了提高和改善 ;缓冲液中阳离子浓度对碱性物质的保留行为有较大的影响 ;碱性物质的最佳流速较中性物质小。由于胺型键合固定相中的疏水烷基链中引入了极性官能团氨基 ,因此碱性物质在该固定相上的行为比在传统的固定相上更为复杂。     

聚N-异丙基丙烯酰胺硅胶键合固定相的应用

采用N-异丙基丙烯酰胺制备了聚N-异丙基丙烯酰胺硅胶键合固定相。试验将聚N-异丙基丙烯酰胺硅胶键合固定相应用于碱性物质和酯类物质的分离,各组分实现了基线分离,且色谱峰形对称;并利用聚N-异丙基丙烯酰胺硅胶键合固定相,在甲醇-水(30+70)混合液为流动相,28~32℃时发生了相转变,由亲水性构象转变为疏水性构象的温敏特性,可将其用于两种极性差异较大的极性化合物分析中。     

Characterization of internal surface reversed-phase silica supports for liquid chromatography

Internal surface reversed-phase (ISRP) supports synthesized from commercially available porous silica particles with a variety of nominal pore diameters and specific surface areas are characterized with regard to physical and chromatographic properties. Bonded phase coverage, pore size, capacity and efficiency measurements are made upon the various ISRP supports in order to evaluate the effect that the physical properties of silica have upon the chromatographic performance of ISRP packings. In addition, various models that describe the pore structure of silica supports are discussed.     

Preparation and investigation of LC packing made by microwave‐assisted solid‐phase synthesis

The preparation of the so‐called bonded phase liquid chromatographic packings is usually carried out by heating the silica, the silane, a catalyst, or a scavenger in an appropriate solvent (often toluene.) Due to the longtime of boiling, the procedure is time and energy consuming, and solvent intensive. The goal of this work is to present a simple, environment‐friendly preparation method with reduced solvent consumption to synthetize RP liquid chromatographic stationary phases. The effects of reaction conditions (amount of reagents, composition of the reagent, microwave energy, reaction time, reproducibility of the synthesis) are discussed. Pore structure, surface coverage, the change of the pore structure and surface coverage upon reaction are demonstrated, the efficiency of the column (van Deemter plot for different solutes) is presented. A variety of applications (aromatic hydrocarbons, halobenzenes, bioactive peptides, resveratrol from red wine) demonstrates the separation power of the new phase.     

Base‐deactivated and alkaline‐resistant chromatographic stationary phase based on functionalized polymethylsilsesquioxane microspheres

Vinyl, chloropropyl, and mercaptopropyl functionalized particles were prepared by a two‐step acidic/alkaline catalyzed co‐hydrolysis/condensation of methyltrimethoxysilane with a different silane precursor that carries chemically reactive functional group including vinyl, chloropropyl, and mercaptopropyl, respectively. The morphology, pore structure, and functional groups of the synthesized packings were studied by SEM, nitrogen adsorption‐desorption measurements, and solid‐state ¹³C ²⁹Si NMR spectroscopy, respectively. The particles show ordered sphere, narrow particle size distribution, and mesoporous structure. The carbon contents of the microspheres are in the range of 17–19%, comparable to those of octadecyl‐bonded silica packings. The three‐kind of microspheres were directly used as packing materials for high‐performance liquid chromatography without size classification. The chromatographic performance of the columns was evaluated and compared with a commercially available C₁₈ phase. The results revealed that these columns possess typical reversed‐phase chromatographic properties with increased hydrophobicity than polymethylsilsesquioxane and symmetric peaks for basic compounds. They were applied to the simultaneous separation of combination bendazol hydrochlorothiazide capsules containing polar and basic drugs with peaks identified by tandem with mass spectrometry. In general, a novel method is provided for the synthesis of different methyltrimethoxysilane‐derived microspheres for high‐performance liquid chromatography, which are advantageous for separating basic compounds.     

硅胶聚合物键合相键合基团及流动相盐浓度和pH值对蛋白质分离的影响

介绍了３种同孔径不同键合基团的大孔径硅胶聚合物键上的合成，３种填料是在１００ｎｍ孔径硅胶上键合乙烯基后，分别与甲基丙烯酸羟乙基酯，二乙烯基苯共聚形成ＨＤＳＩ填料；与甲基丙烯酰胺共聚形成ＰＡＭＳＩ填料；与顺丁烯二酸共聚再与乙二胺反应形成ＰＦＭＳＩ填料。比较了它们对蛋白质分离的特性。     

苯乙烯和马来酸酐包夹硅胶弱阳离子色谱柱的制备及其应用

用马来酸酐包夹硅基制备出一种新型弱阳离子色谱柱填料,它具有良好的色谱性能,可同时分离一价、二价金属阳离子。论述了固定相的制备条件。采用所制备的固定相对热电厂锅炉水样中的阳离子进行了测定。     

Reversed-phase high-performance liquid chromatography of dinucleoside polyphosphates

The reversed-phase high-performance liquid chromatographic separation of purine dinucleoside polyphosphates on octadecyl- and phenyl-bonded silica packings using phosphate-based eluents was studied. The effects of pH, ionic strength and the content of the organic modifiers methanol and acetonitrile in the mobile phase on the retention and other chromatographic parameters are reported. The data obtained were used to establish an isocratic assay for diguanosine and diadenosine polyphosphates.     

Native and modified alumina,titania and zirconia in normal and reversed-phase high-performance liquid chromatography

Summary Chromatographic properties of silica, alumina, titania and zirconia have been investigated in normal phase mode in the separation of test mixtures of basic, neutral and acidic compounds. In contrast to silica the chromatographic behaviour revealed the basic properties of the alumina, titania and zirconia surfaces. Therefore, separation of basic compounds on these packings seems very promising. Lypophilic packings have been synthesized by modification of titania, zirconia and alumina with organosilanes and polymers and tested for the separation of basic compounds and proteins. High hydrolytic stability of the modified packings was observed during separations with strong alkali and acidic eluents.     

Comparative studies of gas chromatographic properties of new packings with chemically bonded complexes

Specific interactions of aliphatic linear and branched hydrocarbons as well as cyclic and aromatic hydrocarbons with new packings containing chemically bonded complexes of transition metals are studied. The possibility of using these packings to separate these compounds is also discussed. The packings under study contain complexes of Cu(II) and Cr(III) chemically bonded to the silica surface. Chlorides of these metals are bonded to the silica surface by the use of the ketoimine group originally from 2-(3-triethoxysililpropylimino)-3-(n-buthyl)-pentanon-4. In order to determine an influence of the performed modification to gas chromatographic properties of the packings, such retention parameters as retention factor, retention index, molecular retention index, and specific retention volume are measured for these compounds. Based on the obtained values, a trial is taken to determine an influence of the nature of the bonded metal from the complex on the retention of the adsorbates under study and a dependence between a structure of an adsorbate molecule and values of charge-transfer interactions with the bonded metal complexes.     

溶质在动态改性氧化锆液相色谱柱上的保留行为

分别采用硬脂酸、环糊精和十二烷基磺酸钠动态改性自制的ZrO2微球,研究了流动相中甲醇和改性剂浓度对苯酚及苯甲酸的衍生物、苯胺衍生物及芳香烃类化合物的色谱保留行为的影响。中性及碱性化合物的保留时间较短,色谱峰对称;酸性化合物保留时间较长,色谱峰拖尾较严重。改性氧化锆表现出反相色谱性能。     

Vergleichende Untersuchungen über stationäre Phasen in der schnellen Flüssig-Chromatographie

The following stationary phase materials for column liquid chromatography: Corasil I and II, Perisorb A, and Zipax were characterized as to their particle size and their pore structure of the porous layer. The superficially porous packings were tested at identical conditions in liquid-solid and liquid-liquid chromatography. The chromatographic parameters k′, h, h=f(u), N /t, K and K · N /t were evaluated. The results were discussed in connection with the surface proporties of the stationary phase materials.