聚(St-DVB)型整体柱的制备及其对蛋白质的色谱分离性能

苯乙烯(St)为单体,二乙烯基苯(DVB)为交联剂,在致孔剂甲苯和十二醇存在下,直接以Ф4.6×50mm色谱柱管为模具,同时在整个反应过程中通过氩气对反应混合物施加一个恒定的压力,通过原位聚合制备了聚(苯乙烯-二乙烯基苯)(PSD)型整体柱。在聚合过程中对反应混合物施加一定的压力,使制备的整体柱表面光滑无凹痕现象,并且使柱子的两头平整不凹陷,有利于提高柱效。本文对整体柱的孔结构及其对蛋白质的分离性能进行了研究。实验结果表明,整体柱内部含有大量类似渠道的大孔,孔径为2μm~3μm,在流速为5ml/min时,背压为7MPa。而且流速对色谱分离效率的影响小,流速从1ml/min增大到3ml/min时,牛血清白蛋白和溶菌酶这两种蛋白质的总分离效能指标K1的值基本保持不变,保持在3.35左右,说明可以通过提高流速实现蛋白质的快速分离。     

牛血清白蛋白修饰毛细管整体柱的制备及组氨酸对映体分离

以甲基丙烯酸缩水甘油酯为单体, 乙二醇二甲基丙烯酸酯为交联剂, 环己醇和正十二醇混合溶液为致孔剂, 在最佳聚合条件下, 以偶氮二异丁腈为引发剂, 制备了毛细管整体柱基质, 并且研究了单体、交联剂及致孔剂对整体柱基质孔结构及渗透性的影响; 使用Epoxy方法在基质表面键合BSA, 制得BSA修饰的毛细管整体柱. 将此毛细管整体柱应用于毛细管电色谱中, 成功地分离出了组氨酸对映体, 分离度良好.     

以聚乙二醇/甲醇为二元致孔剂的新型磺酸甜菜碱型两性离子亲水毛细管整体柱的合成与色谱评价

使用新型二元致孔剂聚乙二醇(PEG)/甲醇,以N,N-二甲基-N-甲基丙烯酰胺基丙基-N,N-二甲基-N-丙烷磺酸内盐(SPP)为单体,季戊四醇三丙烯酸酯(PETA)为交联剂,偶氮二异丁腈(AIBN)为引发剂,通过原位聚合法制备磺酸甜菜碱型两性离子亲水毛细管整体柱。对各反应物的配比进行了优化。结果表明,当单体与致孔剂的质量比为1∶2.5,并且致孔剂中PEG与甲醇的质量比为1∶2,单体内部SPP与PETA的质量比为1∶1,AIBN为总质量的0.1%时为最优配比;PEG/甲醇二元致孔剂的加入实现了对整体柱内部孔径大小的调节,得到了结构更为均一,渗透性、机械稳定性良好的毛细管整体柱,并且理论塔板数与传统制备方法相比有显著提高,在毛细管液相色谱模式下最高可达2.4×105塔板/m。将制备的整体柱应用于毛细管液相色谱和加压毛细管电色谱分离酚类、核苷类等极性小分子混合物,得到了很好的分离效果。     

析因设计法优化制备聚(DVB)型整体柱及分离小分子物质的应用

以二乙烯基苯(DVB)为单体,十二烷醇和甲苯为制孔剂,在引发剂偶氮二异丁腈(AIBN)的作用下,直接以φ4.6×50mm色谱柱为模具,通过原位聚合制备了聚(二乙烯基苯)型整体柱.通过析因设计,以十二烷醇占反应混合物体积比,二乙烯基苯占反应混合物体积比,偶氮二异丁腈所占聚合物质量比3因素,以整体柱对小分子的分离度为响应指标进行了3因素2水平全析因实验.结果表明,引发剂的量对分离度的影响最大,其次为十二烷醇占反应混合物体积比与引发剂占聚合物质量比的交互作用和3因素交互作用,而其他效应,如十二烷醇占反应混合物体积比,DVB占反应混合物体积比,十二烷醇与DVB的交互作用,DVB与引发剂的交互作用的影响较小,可以忽略;从而得到了分离小分子物质的最优配方;通过测定整体柱的比表面积,孔径和孔容分布对其进行表征.所制备的整体柱,能够通过高效液相色谱系统对苯,乙苯和萘这3种小分子物质进行分离.分离条件为:100×4.6mm I.D.色谱柱,流动相为乙腈:水=80:20(v/v),流速1mL/min,检测峰值254nm.色谱柱背压5MPa,分离时间10min.     

正交试验法优化制备用于小分子物质分离的聚(甲基丙烯酸缩水甘油酯-二乙烯基苯)型整体柱

以甲基丙烯酸缩水甘油酯(GMA)为单体,二乙烯基苯(DVB)为交联剂,以环己醇和正十二醇为致孔剂,过氧化苯甲酰(BPO)为引发剂,直接以50 mm×4.6 mm色谱柱为模具,通过原位聚合制备聚(GMA-DVB)型整体柱。以GMA和DVB的体积比、环己醇和十二醇的体积比以及BPO占聚合物的质量分数为三因素,以分离苯和乙苯等小分子物质时的半峰宽分离度(R1/2)为考察指标,进行三因素三水平的正交试验,通过测定整体柱的比表面积、孔径和孔容分布对其进行表征。结果表明,制备整体柱的最优配方为V(GMA): V(DVB): V(环己醇): V(正十二醇)=0.825:0.825:1.32:2.03, BPO的质量分数为0.7%。应用所制备的整体柱分离苯和乙苯等小分子物质,理论塔板数达到37000塔板/m, R1/2值达到7.14,完全达到基线分离,分离时间小于10 min。该方法制备整体柱的重复性好,柱效较高,基本满足商品化要求。     

磺酸甜菜碱型两性离子亲水毛细管整体柱制备及应用

以3-[N,N-二甲基-[2-(2-甲基丙-2-烯酰氧基)乙基]铵]丙烷-1-磺酸内盐(SPE)为功能单体,季戊四醇三丙烯酸酯(PETA)为交联剂,环己醇和乙二醇(EG)为致孔剂,通过原位聚合法制备磺酸甜菜碱型两性离子亲水毛细管整体柱。优化单体、交联剂和致孔剂的比例等因素,考察了不同SPE含量对整体柱性能和选择性的影响。在最优制备条件下,以苯酚类化合物、烷基苯类化合物和苯甲酸类化合物为分离对象,评价该整体柱的色谱性能以及分离机理。在不同的色谱条件下,该整体柱具有亲水、疏水以及离子交换作用。此整体柱在0.05 m L/min的流速下(线速度为0.265 mm/s)分离烷基苯类化合物时,柱效高达41000~56000 plates/m,该整体柱重现性良好,连续运行的重现性(RSD)低于1.2%。在亲水/离子交换色谱模式下,该整体柱可应用于核苷和碱基的高效分离。     

反相毛细管整体柱的制备及其在多肽混合物分离中的应用

采用甲基丙烯酸月桂酯为基础功能单体,乙二醇二甲基丙烯酸酯为交联剂,正十二醇、1,4-丁二醇及二甲基亚砜为致孔剂,在内径为75 μm的石英毛细管内制备了具有良好机械性能及化学稳定性的反相毛细管整体柱。考察了致孔剂的种类、比例以及交联剂在单体混合物中的比例对柱压和分离效果的影响;以单体15%、交联剂15%、致孔剂70%(均为质量分数)作为优化配方,在70 ℃条件下反应24 h;并对所合成的毛细管整体柱进行了电镜表征,测试了流速、柱长与柱压的关系。结果表明,毛细管整体柱的通透性良好,可通过延长柱长的方法提高分离效果。将所制备的毛细管整体柱装于纳升级高效液相色谱仪上进行牛血清白蛋白及血浆样本的胰蛋白酶酶切液的分离,获得了比较理想的分离效果。     

乙烯基酯树脂整体柱用于溶菌酶的分离

以甲基丙烯酸间苯二酚二缩甘油酯树脂兼作单体和交联剂,偶氮二异丁腈(AIBN)为引发剂,十六醇为致孔剂,在不锈钢柱管中制成聚合物整体柱,通过电镜分析考察了该聚合物的微观孔结构.以此聚合物整体柱为基质,将该柱修饰为具有二醇基的聚合物整体柱,考察了此修饰柱的通透性,并将其作为高效液相色谱固定相考察了该柱对溶菌酶(Lys)的最大吸附量、α-淀粉酶(α-Amy)和溶菌酶在该柱上的分离,并对蛋清中的溶菌酶进行了分离.结果表明,分离效果良好,为蛋白分离提供了一种简单、快捷、有效的方法.     

士的宁分子印迹整体柱的制备

以士的宁为模板分子，甲基丙烯酸为功能单体，乙二醇二甲基丙烯酸酯作为交联剂，甲苯和十二醇混合溶液为致孔剂，用原位分子印迹技术，合成了士的宁分子印迹整体柱。通过优化合成条件，结果显示：模板分子、功能单体与交联剂之间的比例以1：4：16最佳，致孔剂中甲苯的最佳含量为18％（V/V)；对士的宁整体柱的色谱条件包括流动相组成、流速、柱温等进行了考察，并用于士的宁和马钱子碱的分离，其分离因子为3．5。     

以金纳米溶胶为中间体同时制备阳离子及反相模式整体柱

本文以甲基丙烯酸缩水甘油酯(GMA)为单体,二甲基丙烯酸乙二酯(EDMA)为交联剂,环己醇和十二醇为致孔剂,过氧化苯甲酰(BPO)为引发剂,制备了有机聚合整体材料基质,利用金纳米粒子(AuNPs)易与巯基基团(-SH)共价键合的特点和有机聚合整体柱的优点,以半胱胺和金纳米溶胶(AuNPs)为中间体,依次采用2-巯乙基磺酸钠和正十八烷硫醇修饰该柱,制备了表面具有阳离子交换和疏水作用两种模式的整体柱,拓宽了有机聚合物整体柱的应用领域。制备的整体柱适用于不同色谱模式下蛋白质或多肽混合物的分离。     

Preparation and evaluation of a neutral methacrylate-based monolithic column for hydrophilic interaction stationary phase by pressurized capillary electrochromatography

A polar and neutral polymethacrylate-based monolithic column was evaluated as a hydrophilic interaction capillary electrochromatography (HI-CEC) stationary phase with small polar–neutral or charged solutes. The polar sites on the surface of the monolithic solid phase responsible for hydrophilic interactions were provided from the hydroxy and ester groups on the surface of the monolithic stationary phase. These polar functionalities also attract ions from the mobile phase and impart the monolithic solid phase with a given zeta potential to generate electro-osmotic flow (EOF). The monolith was prepared by in situ copolymerization of a neutral monomer 2-hydroxyethyl methacrylate (HEMA) and a polar cross-linker with hydroxy group, pentaerythritol triacrylate (PETA), in the presence of a binary porogenic solvent consisting cyclohexanol and dodecanol. A typical HI-CEC mechanism was observed on the neutral polar stationary phase for both neutral and charged analytes. The composition of the polymerization mixture was systematically altered and optimized by altering the amount of HEMA in the polymerization solution as well as the composition of the porogenic solvent. The monoliths were tested in the pCEC mode. The resulting monoliths had different characteristics of hydrophilicity, column permeability, and efficiency. The effects of pH, salt concentration, and organic solvent content on the EOF velocity and the separation of nucleic acids and nucleosides on the optimized monolithic column were investigated. The optimized monolithic column resulted in good separation and with greater than 140,000 theoretical plates/m for pCEC.     

Novel Polymer Monolithic Column for Hydrophilic Compounds

A hydrophilic polymer-based monolithic column was prepared from water-soluble crosslinking agents for liquid chromatography. The column media were prepared with a diacrylate monomer, aqueous polyethylene glycol, methanol, and a water-soluble radical initiator. A fused silica capillary column, which was significantly modified by vinyl groups, was utilized as the outer column in order to control polymer shrinkage. We optimized the modification of the capillary column, composition of the monomer and porogen, and polymerization condition. The optimized column exhibited high hydrophilicity and achieved the baseline separation of nucleobases by using only a buffered solution as the mobile phase. Additionally, another column prepared from binary water-soluble crosslinking agents exhibited lower nonspecific adsorption of several proteins. Basically, we would show the possibility of a new type of polymer monolith prepared from water-soluble crosslinker and water-soluble porogenic solvent, and they can be used for chromatographic separation without non-specific hydrophobic interaction.     

Preparation and characterization of a molecularly imprinted monolithic column for pressure‐assisted CEC separation of nitroimidazole drugs

A polymethacrylate‐based molecularly imprinted monolithic column bearing mixed functional monomers, using non‐covalent imprinting approach, was designed for the rapid separation of nitroimidazole compounds. The new monolithic column has been prepared via simple in situ polymerization of 2‐hydroxyethyl methacrylate, dimethylaminoethyl methacrylate and ethylene dimethacrylate, using (S)‐ornidazole ((S)‐ONZ) as template in a binary porogenic mixture consisting of toluene and dodecanol. The composition of the polymerization mixture was systematically altered and optimized by altering the amount of monomers as well as the composition of the porogenic solvent. The column performance was evaluated in pressure‐assisted CEC mode. Separation conditions such as pH, voltage, amount of organic modifier and salt concentration were studied. The optimized monolithic column resulted in excellent separation of a group of structurally related nitroimidazole drugs within 10 min in isocratic elution condition. Column efficiencies of 99 000, 80 000, 103 000, 60 000 and 99 000 plates/m were obtained for metronidazole, secnidazole, ronidazole, tinidazole and dimetridazole, respectively. Parallel experiments were carried out using molecularly imprinted and non‐imprinted capillary columns. The separation might be the result of combined effects including hydrophobic, hydrogen bonding and the imprinting cavities on the (S)‐ONZ‐imprinted monolithic column.     

新型有机-无机杂化整体柱的制备及其在毛细管液相色谱及加压毛细管电色谱中的应用

以丙烯酰胺(AM)为单体,八乙烯基倍半硅氧烷(POSS)为交联剂,偶氮二异丁腈(AIBN)为引发剂,四氢呋喃(THF)为致孔剂,通过原位聚合法制备了poly(POSS-co-AM)有机-无机杂化整体柱,并对各反应物的配比进行了优化。结果表明,当功能单体与致孔剂、POSS与AM的质量比均为1.0: 5.0, AIBN的质量分数为0.1%时,杂化整体柱的柱效最高。无机材料的引入使整体柱结构均匀并具有良好的渗透性,该整体柱既能用于亲水色谱模式,也能用于反相色谱模式。将制备的整体柱用于毛细管液相色谱和加压毛细管电色谱分离核苷类、胺类、硝基苯胺类等化合物,获得了良好的效果。     

微电渗流毛细管原位柱的制备及性能考察

 采用二元致孔剂原位聚合的方法制备了一种新型微电渗流毛细管原位柱。与三元致孔剂制柱方法相比 ,具有制备过程简单、重复性好、能够方便地通过改变致孔剂配比来改变柱床的孔径和孔结构的特点。得到的毛细管柱内部结构均匀 ,通透性好。通过对改变不同致孔剂配比所制备的原位柱的孔结构特征及电渗流情况考察 ,及对柱长和柱径与电渗流的关系的探讨 ,发现制备的原位柱在较高 pH值和较高的有机改性剂浓度条件下 ,电渗流均能保持在较低值 ,可以适应不同电泳分离模式的需要。     

甲基丙烯酸基质的毛细管电色谱整体柱的制备与应用

以甲基丙烯酸(MAA)为功能单体,同时又作电渗流改性剂, 乙二醇二甲基丙烯酸酯(EDMA)为交联剂,甲苯和异辛烷为致孔剂,Irgacure 1800为光引发剂,采用紫外光引发原位聚合反应制备出毛细管电色谱整体柱.对影响电渗流的因素如pH、乙腈含量及离子强度等进行了讨论.使用制备的整体柱在3种模式(加压电色谱、气压驱动和电色谱)下对7种中性物质(硫脲、苯、甲苯、乙苯、萘、联苯和菲)的混合物实现基线分离,同时还可实现酸性物质(邻羟基苯甲酸、苯甲酸、苯乳酸、扁桃酸)和碱性物质(苯胺、甲苯胺、乙酰基苯胺和N-甲基苯胺)的快速分离.     

Preparation and evaluation of rigid porous polyacrylamide-based strong cation-exchange monolithic columns for capillary electrochromatography

A CEC monolithic column with strong cation-exchange (SCX) stationary phase based on hydrophilic monomers was prepared by in situ polymerization of acrylamide, methylenebisacrylamide, and 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) in a complete organic binary porogenic solvent consisting of DMSO and dodecanol. The sulfonic groups provided by the monomer AMPS on the surface of the stationary phase generate an EOF from anode to cathode, and serve as an SCX stationary phase at the same time. The monolithic stationary phase exhibited normal-phase chromatographic behavior for neutral analytes. For charged analytes, electrostatic interaction/repulsion with the monolith was observed. The strong SCX monolithic column has been successfully employed in the electrochromatographic separation of basic drugs, peptides, and alkaloids extracted from natural products.     

大孔交联聚甲基丙烯酸羟乙酯树脂的制备及其结构性能

双甲基丙烯酸乙二醇酯;大孔树脂;孔结构;大孔交联聚甲基丙烯酸羟乙酯树脂的制备及其结构性能