Poly(glycerin 1,3‐dimethacrylate)‐based monolith with a bicontinuous structure tailored as HPLC column by photoinitiated in situ radical polymerization via viscoelastic phase separation

In this article we described our new approach to the polymer monolith with its morphology tailored for HPLC application to small solutes such as drug candidates. We prepared polymer monoliths based on glycerin 1,3‐dimethacrylate, GDMA with a bicontinuous structure by in situ photoinitiated free radical polymerization (UV irradiation at 365 nm). Our photopolymerization was carried out with a monodispese ultra high molecular weight polystyrene solution in chlorobenzene uniquely formulated as a porogen. The poly‐GDMA monoliths in bulk, rod and capillary thus prepared showed a bicontinuous network‐like structure featured by their fine skeletal thickness nearly in sub μm size. This monolithic structure was considered as a time‐evolved morphology frozen by UV‐irradiation via viscoelastic phase separation induced by the said porogenic polystyrene solution. According to our μHPLC measurement with acetophenone as a model solute, the UV prepared poly‐GDMA capillary demonstrated a much shaper elution profile affording higher column efficiency and permeability as compared with the thermally prepared capillary of the same bore size. Our investigation showed experimentally that poly‐GDMA monoliths with a well‐defined bicontinuous structure could be prepared reproducibly by photoinitiated radical polymerization via viscoelastic phase separation using the said unique porogen. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 4651–4673, 2008     

2,4,6‐trichlorophenyl acrylate emulsion‐templated porous polymers (PolyHIPEs). Morphology and reactivity studies

A series of monolithic crosslinked polymers with 2,4,6‐trichlorophenyl acrylate as a reactive component was prepared by free radical polymerization of the internal phases of high internal phase emulsions (HIPEs). The volume ratio of water to oil phase (void volume; 60–90%), crosslinker type (divinylbenzene or ethylenglycol dimethacrylate) and quantity (30–50 mol %) and type of porogenic solvent (chlorobenzene, toluene, chloroform, dichloroethane) were altered to study these effects on the structure and reactivity of the monolithic polymers. The polymer supports were characterized by scanning electron microscopy (SEM), FTIR spectroscopy, elemental analysis and mercury intrusion porosimetry. SEM images revealed an open cellular structure with voids between 1 and 12 μm and window sizes between 0.3 and 3 μm. The porogen had an influence on the surface area, being larger with added porogen and the influence being highest with toluene. Adding toluene also influenced the void size, increasing the average diameter from ～2 μm (no porogen) to ～12 μm (added toluene). Monolithic supports were functionalized by reaction of the ester moieties with tris(2‐aminoethyl)amine derivative and by hydrolysis of the ester groups to carboxylic acids. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 4043–4053, 2007     

Porogen formulations for obtaining molecularly imprinted polymers with optimized binding properties

This paper examines the formulation of new porogenic mixtures used to prepare molecularly imprinted polymers (MIPs) in both thin film and bulk monolith formats. Films were cast by using spin coating to spread a pre-polymerization mixture onto a substrate, and rapid curing of the films was achieved with UV photolysis. The use of a low volatility solvent in combination with a linear polymer porogen resulted in a porous morphology and a 60-fold enhancement in the binding capacity, relative to a non-porous film prepared with a highly volatile solvent and in the absence of the polymer porogen. The opposite effect was seen in MIPs that were prepared in the traditional bulk monolith format, for which the binding efficiency of the MIP decreased monotonically with the concentration of the linear polymer porogen. Furthermore, bulk MIPs that were prepared in the presence of linear polymer porogens exhibited significantly decreased specific surface areas (from 620 to 8 m²/g for samples prepared with pure solvent and 50% polymer porogen, respectively). Despite the change in binding capacity and morphology, the selectivity of the bulk MIPs remained unaffected by the presence of the polymer porogens (approximately 50% chiral selectivity for all bulk MIPs considered). This difference in behavior of the two systems was attributed to the large difference in the kinetics of polymerization.     

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.     

Effect of the nature of a porogen on the porous structure of monolithic polydivinylbenzene sorbents

In quartz capillaries, macroporous monolithic sorbents based on divinylbenzene are synthesized and their porous structure is studied via inverse size-exclusion-hydrodynamic chromatography. Either a single-component porogen (a higher alcohol) or a two-component porogen (a mixture of a higher alcohol and mesitylene) is used for the synthesis of monoliths. The removal of a solvent good for a polymer from a porogen results in an increase in the size of flow-through channels and a decrease in the free-space volume inside the monolith; this space is used for the separation of polymer sorbates (the working volume of a column). At the same time, the volume of micro- and mesopores in the monolith structure is practically independent of the content of the good solvent in the porogen. It is inferred that the good porogen plays an active role in formation of the macroporous structure of monoliths. The structure of monoliths obtained on the basis of the two-component porogen with the use of nonanol and mesitylene or toluene is optimum for the molecular-mass analysis of polymers.     

Development of rigid biporous polymeric adsorbent for protein chromatography

Summary A novel biporous poly (glycidyl methacrylate-triallyl isocyanurate-divinylbenzene) resin (denoted as Resin C) was prepared byin-situ co-polymerization with both superfine granules of sodium sulfate and the solvents toluene and n-heptane as porogenic agents. This material is based on a novel porogenic mode of a combination of both solid granules and solvents. The properties of Resin C were characterized, and then compared with both Resin A (where only solvents as porogen) and Resin B (where only solid granules are used as porogen). The biporous resin showed good mechanical performance, high dynamic adsorption capacity and column efficiency at high flow-rates. These factors indicate the biporous resin as a promising adsorbent for high-speed protein separation by chromatography.     

Fast preparation of polystyrene-based monolith using microwave irradiation for micro-column separation

The conventional analytical HPLC was successfully developed for micro-column separation by using a simple eluate splitting system, a self-preparation of monolithic column and an on-capillary column detector in our laboratory. A typical polystyrene-based monolith was quickly prepared inside the fused-silica capillary, which in situ polymerization was carried out in 10 min by microwave irradiation. The reactant solution consisted of styrene (ST) as a functional monomer, divinylbenzene (DVB) as a cross-linking agent, toluene and isooctane as porogenic solvents, and azobisisobutyronitrile (AIBN) as an initiator. The monolith was proved to form in the center of the capillary and adhered to the column inner wall by the scanning electron micrograph. Its chromatographic behaviors were evaluated in detail by varying the flow rate and percentage of mobile phases, and under the optimal condition, baseline separation of the model analytes including thiourea, benzene, toluene, ethylbenzene was obtained with a highest theoretical plate number near 11,290 N/m by the developed capillary HPLC. Furthermore, the stability and porosity of the prepared monolith were systematically investigated by a simple flow method. Figure A polystyrene-based monolith was rapidly prepared inside the fused-silica capillary, which in situ polymerization was carried out about 10 min by microwave irradiation.     

Characteristics and fluidic properties of porous monoliths prepared by radiation-induced polymerization for Lab-on-a-Chip applications

Porous polymer monoliths were prepared by UV- or EB-induced polymerization of hydroxyethyl methacrylate (HEMA) and ethylene dimethacrylate (EDMA) as network precursors dissolved in porogenic solvent mixtures composed of methanol and n-hexane. The fluidic properties and the pressure resistance of porous monoliths synthesized into 1 mm i.d. capillaries and in 100 μm-wide microchannels were investigated. The influence of photopolymerization time (or electron beam dose) and monomer content on flow properties is discussed on the basis of morphological features. The two types of radiation can be used to achieve the in situ fabrication of monolith inside microsystems. The permeability of the porous monoliths can be adjusted by tuning compositional and processing parameters.     

Preparation and evaluation of hydroxylated poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolithic capillary for in-tube solid-phase microextraction coupled to high-performance liquid chromatography

A hydroxylated poly(glycidyl methacrylate-co-ethylene dimethacrylate) (GMA-co-EDMA) monolithic capillary was prepared and investigated for in-tube solid-phase microextraction (SPME). The polymer monolith was synthesized by in-situ polymerization of GMA and EDMA in the presence of dodecanol and toluene as the mixed porogenic solvents. After polymerization, glycidyl groups were hydrolyzed with sulfuric acid to produce diol groups at the surface of the porous monolith. To investigate the extraction mechanism, several groups of model analytes (including neutral, acidic and basic) were selected to perform extractions. The resulting monolith showed high extraction selectivity towards polar compounds, which resulted from the enhancement of dipole-dipole and hydrogen bonding interactions relative to hydrophobic interactions. The equilibrium extraction time profiles were also monitored for those model compounds to assess the extraction capacity of the monolithic capillary. Moreover, the hydroxylated poly(GMA-co-EDMA) monolithic capillary exhibited satisfactory reproducibility and stability. Finally, the in-tube SPME-HPLC method, based on the developed monolithic capillary as the extraction media, was successfully applied to the determination of five polar organic contaminants in lake water.     

Enantioseparation of 2-aryloxypropionic acids on chiral porous monolithic columns by capillary electrochromatography. Evaluation of column performance and enantioselectivity

The enantioseparation of 2-aryloxypropionic acids by capillary electrochromatography was tested on columns with a monolithic stationary phase prepared from silanized fused-silica capillaries (100 microm I.D.) by in situ copolymerization of glycidyl methacrylate, ethylene glycol dimethacrylate and methyl methacrylate in the presence of formamide and 1-propanol as the porogen solvents. The porous chiral monolithic stationary phases were prepared by reaction of the epoxy-groups at the surface of the monolith with (+)-1-(4-aminobutyl)-(5R,8S,10R)-terguride. To attain the minimum HETP values for the enantiodiscrimination of 2-phenoxypropionic acid, the influence of the composition of polymerization solution on column total porosity and efficiency was investigated. Optimum mobile phase conditions were found for all analytes tested using acetonitrile-methanol mixtures containing triethylamine and acetic acid as the buffer components. Furthermore, the chemical and mechanical stabilities of the columns were satisfactory, allowing hundreds of analyses.     

Preparation and evaluation of a phenylboronate affinity monolith for selective capture of glycoproteins by capillary liquid chromatography

A phenylboronate affinity monolith was prepared and applied to the selective capture of glycoproteins from unfractionated protein mixtures. The monolith was synthesized in a 100 μm i.d capillary by an in situ polymerization procedure using a pre-polymerization mixture consisting of 4-vinylphenylboronic acid (VPBA) as functional monomer, ethylene dimethacrylate (EDMA) as crosslinker, diethylene glycol and ethylene glycol as binary porogenic solvents, and azobisisobutyronitrile (AIBN) as initiator. The prepared monolith was characterized in terms of the morphology, pore property, and recognition property. The selectivity and dynamic binding capacity were evaluated by using standard glycoproteins and nonglycoproteins as model proteins. The chromatographic results demonstrated that the phenylboronate affinity monolith had higher selectivity and binding capacity for glycoprotein than nonglycoprotein. The resulting phenylboronate affinity monolith was used as the sorbent for in-tube solid phase microextraction (in-tube SPME), and the extraction performance of the monolith was assessed by capture of ovalbumin from egg white sample.     

Chiral separation of binaphthol enantiomers on molecularly imprinted polymer monolith by capillary electrochromatography.

A novel enantioseparational monolithic stationary phase for binaphthol based on a molecular imprinting method was introduced and evaluated in capillary electrochromatography (CEC). The monolithic stationary was prepared by the in situ copolymerization of methacrylic acid and ethylene glycol dimethacrylate in a porogenic solvent (toluene or toluene-isooctane) in the presence of an imprinting molecule, (R)-1,1'-bi-2,2'-naphthol. Such stationary phases could separate the enantiomers of binaphthol. The influence of several parameters on the column permeability was investigated. These parameters included the polymerization time, the molar ratio of the functional monomer to the imprinting molecule and the content of porogen. The influence of the polymerization condition and the electrochromatographic parameters on the enantiomer separation was also studied. Initial studies showed that a higher molecular ratio of the imprinted molecule to the functional monomer, a higher content of porogen, a higher content of acetonitrile, a higher pH, as well as the addition of Tween 20, gave a higher enantiomer selectivity.     

二苯甲酰-D-酒石酸分子印迹整体柱的制备及其色谱性能

采用紫外光谱法选择丙烯酰胺为功能单体,以二苯甲酰-D-酒石酸(D-DBTA)为模板分子,丙烯酰胺为功能单体,二甲基丙烯酸乙二醇酯为交联剂,甲苯和十二醇为混合致孔剂,用原位分子印迹技术,合成了D-DBTA分子印迹整体聚合物。在优化色谱条件的基础上,25min内基本实现了DBTA消旋体的手性分离,分离度达1.25。对色谱分离过程中的热力学进行分析,在所考察的温度范围内,色谱分离存在两种不同的热力学过程,且两者与范特霍夫方程能够很好地吻合。在这两种热力学过程的转换温度处,分离因子α达最大值。     

Narrowly dispersed molecularly imprinted microspheres prepared by a modified precipitation polymerization method

A modified precipitation polymerization (MPP) method was established to prepare narrowly dispersed molecularly imprinted polymeric microspheres. MPP was stabilizer and surfactant free and needed only small amount of porogen (about 50 wt.%). Only part of alcohols and all the alkanes tested formed particles. Using a mixture of alkane and toluene as porogen, the carbon numbers of alkanes and solubility parameter of porogenic solvents were important factors in controlling particle morphology. Nearly mono-dispersed microspheres with diameter of about 2-3 μm were synthesized by MPP using mineral oil:toluene = 2:3 as porogen. Template did not affect the formation of globe microspheres in MPP. Microspheres prepared under the lowest reaction temperature had the highest binding capability. When used as sorbents of high performance liquid chromatography (HPLC), the microspheres prepared by MPP using bisphenol A, estradiol, and tebuconazole as template had similar binding selectivity and higher binding capability compared to microspheres synthesized by classical precipitation polymerization. Photoinitiation and low reaction temperature were important factors attributed to better binding capability of microspheres prepared by MPP.     

Fine control of the porous structure and chromatographic properties of monodisperse macroporous poly(styrene-co-divinylbenzene) beads prepared using polymer porogens

The porous structure of monodisperse macroporous beads can be controlled by using soluble polymers with well-defined structural characteristics as part of the porogenic mixture. In general, the use of linear polystyrene as a porogen in the preparation of poly (styrene-co-divinylbenzene) beads shifts the pore size distribution towards larger pores. While a direct correlation between pore size and molecular weight of the porogen has been established, the chemical composition of the polymer porogen has no effect on the porous and chromatographic properties of the beads. These findings suggest that the average molar volume of the porogenic system is important while the miscibility of the polymer porogen with the crosslinked polymer that is formed is of little relevance. © 1994 John Wiley & Sons, Inc.     

Retentivity and enantioselectivity of uniformly-sized molecularly imprinted polymers for (S)-nilvadipine in aqueous and non-aqueous mobile phases.

Uniformly-sized molecularly imprinted polymers (MIPs) for (S)-nilvadipine have been prepared by a multi-step swelling and polymerization method using methacrylic acid or 4-vinylpyridine (4-VPY) as a functional monomer, ethylene glycol dimethacrylate (EDMA) as a cross-linker, and toluene, chloroform, cyclohexanol or phenylacetonitrile as a porogen. The chiral recognition abilities of the MIPs for nilvadipine were evaluated using aqueous and non-aqueous mobile phases. Among the MIPs, the (S)-nilvadipine-imprinted 4-VPY-co-EDMA polymers prepared using toluene as a porogen showed the highest recognition ability for nilvadipine in both aqueous and non-aqueous mobile phases. In addition to molecular shape recognition, hydrogen-bonding interactions of the NH proton of nilvadipine with a pyridyl group of the (S)-nilvadipine-imprinted 4-VPY-co-EDMA polymers could play an important role in the retention and chiral recognition of nilvadipine in aqueous and non-aqueous mobile phases. Furthermore, the MIP for (S)-nilvadipine gave the highest molecular recognition ability when a porogenic solvent during polymerization was used as the mobile phase modifier.     

Preparation of hemispherical particles by cleavage of micrometer-sized,spherical poly(methyl methacrylate)/polystyrene composite particle with Janus structure: effect of molecular weight

Micrometer-sized, hemispherical polymer particles were prepared as a result of cleavage of spherical Janus poly(methyl methacrylate) (PMMA)/polystyrene (PS) composite particle by treating particles with acetone/water solutions. The original PMMA/PS composite particles were prepared by the slow evaporation of toluene from homogeneous PMMA/PS/toluene droplets dispersed in aqueous solution of sodium dodecyl sulfate in advance. Appropriate molecular weights of PMMA and PS were necessary for occurrence of the cleavage of the Janus composite particle, resulting in PMMA and PS hemispherical particles. The cleavage depended on the composition of the acetone/water solution, which was explained by selective solvent absorption into the polymer phases. The results strongly support the cleavage mechanism of Janus composite polymer particles that had been proposed earlier.     

光聚合整体式咖啡因印迹毛细管柱的制备及分离性能

分子印迹技术作为一种制备对目标分子具有专一识别能力的功能高分子的方法 ,近年来在化学化工、生物化学与生物技术的许多领域中得到广泛应用 [1～ 4 ] .分子印迹技术与微分离方法 (包括微柱液相色谱、毛细管电泳、毛细管电色谱和芯片分离等 )结合已引起人们极大的兴趣和关注[5,6] .毛细管柱是毛细管电色谱和微柱液相色谱的关键部件 ,目前普遍使用的是烷基键合硅胶微粒的填充柱 ,存在制备时须烧塞和填充两大困难 ,以及使用时易产生气泡和易折断等缺点 .将含被识别分子 (印迹分子 )、交联剂、溶剂、功能单体和引发剂的混合液注入毛细管 ,经光…     

Poly(glycidyl methacrylate‐co‐N‐methylolacrylamide‐co‐ethylene dimethacrylate) monolith coupled to high‐performance liquid chromatography for the determination of adenosine phosphates in royal jelly

A polymer monolith microextraction method coupled with high‐performance liquid chromatography was developed for the determination of adenosine triphosphate, adenosine diphosphate, and adenosine monophosphate. The monolithic column was synthesized inside fused‐silica capillaries using thermal initiation free‐radical polymerization with glycidyl methacrylate as the monomer, ethylene dimethacrylate as the cross‐linker, cyclohexanol, and 1‐dodecanol as the porogen. N‐Methylolacrylamide, an important hydrophilic monomer, was incorporated into the polymerization mixture to enhance the hydrophilicity of the poly(glycidyl methacrylate‐co‐ethylene dimethacrylate) column. The obtained poly(glycidyl methacrylate‐co‐N‐methylolacrylamide‐co‐ethylene dimethacrylate) monolith was characterized by scanning electron microscopy, Fourier‐transform infrared spectra, and X‐ray photoelectron spectroscopy. Optimum conditions for the preconcentration and separation of the target adenosines were also investigated. Under the optimum conditions, we obtained acceptable linearities, low limits of detection, and good relative standard deviations. The developed polymer monolith microextraction with high‐performance liquid chromatography method exhibited a good performance with recovery values in the range of 76.9?104.7% when applied to the determination of the adenosines in five royal jelly samples.     

Novel method to prepare polystyrene-based monolithic columns for chromatographic and electrophoretic separations by microwave irradiation

Microwave irradiation can provide a viable alternative to the traditional means such as ultraviolet light and thermal initiation for the preparation of monolithic capillary columns. Polystyrene-based monolithic stationary phases were prepared in situ in fused-silica capillaries and simultaneously in vials. The column permeability, electrophoretic and chromatographic behavior were evaluated using pressure-assisted capillary electrochromatography (pCEC), capillary electrochromatography (CEC) and low pressure liquid chromatography (LPLC). With an optimal monolithic material, the largest theoretical plates for preparing the column could be close to 18,000 plates/m for thiourea in the mode of pCEC. Furthermore, the influence of the composition of the porogenic solvents (toluene/isooctane) on the morphology of organic-based monoliths [poly(styrene-divinylbenzene-methacrylic acid)] was systematically studied with mercury intrusion porosimetry and scanning electron microscopy. The monoliths which were prepared with a high content of isooctane had a bigger pore size and better permeability, and hence resulted in a faster separation.