Characterization of several stationary phases prepared by thermal immobilization of poly(methyltetradecylsiloxane) onto silica surfaces

Variations of a thermal immobilization procedure using poly(methyltetradecilsiloxane) and silica produced fourteen stationary phases with carbon contents of 4-18%. The stationary phases were chromatographically evaluated with the Engelhardt, SRM 870 and Tanaka tests. Classifications using USP and Euerby procedures indicate that the new immobilized phases are different from most commercial phases although there was some similarity with phases that have high ion-exchange interactions. The retention mechanism involved in the separation of basic solutes on several of the new stationary phases was studied by varying pH, type of Lewis base and the ionic strength of the eluent. The separations are strongly influenced by the chemistry of the accessible free silanols. The stationary phases present good selectivity at intermediate pH where the basic analytes were protonated, suggesting use of intermediate pH for these separations. Stability tests show that the stationary phases have poor stability at very high pH, even at 23°C, but good stability in acidic mobile phases, even at 75°C, as expected for an immobilized polymer stationary phase.     

New poly(ionic liquid)-grafted silica multi-mode stationary phase for anion-exchange/reversed-phase/hydrophilic interaction liquid chromatography

A new poly(ionic liquid)-grafted silica stationary phase was prepared and characterized. It was then applied for multi-mode chromatographies, including ion-exchange, reversed-phase, and hydrophilic interaction liquid chromatographies for the effective separation of anions, hydrophobic compounds, and small polar molecules, respectively.     

Chromatographic evaluation of reversed-phase/anion-exchange/cation-exchange trimodal stationary phases prepared by electrostatically driven self-assembly process

This work describes chromatographic properties of reversed-phase/cation-exchange/anion-exchange trimodal stationary phases. These stationary phases were based on high-purity porous spherical silica particles coated with nano-polymer beads using an electrostatically driven self-assembly process. The inner-pore area of the material was modified covalently with an organic layer that provided both reversed-phase and anion-exchange properties while the outer surface was coated with nano-sized polymer beads with strong cation-exchange characteristics. This design ensured spatial separation of the anion-exchange and the cation-exchange regions, and allowed reversed-phase, anion-exchange and cation-exchange retention mechanisms to function simultaneously. Chromatographic evaluation of ions and small molecules suggested that retention of ionic analytes was influenced by the ionic strength, pH, and mobile phase organic solvent content, and governed by both ion-exchange and hydrophobic interactions. Meanwhile, neutral analytes were retained by hydrophobic interaction and was mainly affected by mobile phase organic solvent content. Depending on the specific application, selectivity could be optimized by adjusting the anion-exchange/cation-exchange capacity ratio (selectivity), which was achieved experimentally by using porous silica particles with different surface areas.     

High-performance liquid chromatographic stationary phases based on poly(dimethylsiloxane) immobilized on silica

This work describes the preparation and characterization of six stationary phases for high-performance liquid chromatography (HPLC) obtained by deposition of poly(dimethylsiloxane) (PDMS) in HPLC silica particles, followed by immobilization using different processes (thermal treatments, thermal treatment + microwave irradiation, self-immobilization + gamma irradiation and self-immobilization + microwave irradiation). The chromatographic parameters of all the phases were evaluated with a mixture of test compounds having varied natures (acid, basic and neutral). The stability of one of these phases was evaluated in both a neutral mobile phase and a higher pH mobile phase used at an elevated temperature, with promising results.     

HPLC separation of acetaminophen and its impurities using a mixed-mode reversed-phase/cation exchange stationary phase

Determination of acetaminophen and its main impurities: 4-nitrophenol, 4'-chloroacetanilide, as well as 4-aminophenol and its degradation products, p-benzoquinone and hydroquinone has been developed and validated by a new high-performance liquid chromatography method. Chromatographic separation has been obtained on a Hypersil Duet C18/SCX column, using gradient elution, with a mixture of phosphate buffer (pH = 4.88) and methanol as a mobile phase. Analysis time did not exceed 14.5 min and good resolutions, peak shapes and asymmetries have resulted. The linearity of the method has been tested in the range of 5.0-60 μg/mL for acetaminophen and 0.5-6 μg/mL for the other compounds. The limits of detection and quantification have been also established to be lower than 0.1 μg/mL and 0.5 μg/mL, respectively. The method has been successfully applied for the analysis of commercial acetaminophen preparations.     

NMR measurement of the surface dynamics of poly(dimethylsiloxane) adsorbed on silica gel

CPMAS-DD ¹³C NMR spectroscopy was used to examine the mobility of poly(dimethylsiloxane) adsorbed on silica gel (PDMS/SiO₂) at submonolayer coverages. The spin-lattice relaxation time in the rotating frame (T_1ρH) decreased linearly with increasing loading. This is consistent with a decrease in the mobility of the polymer segments as the loading is increased. The decrease in mobility results from interpolymer interference. We propose a model that explains these results in terms of a surface intrinsic viscosity that incorporates the polymer-polymer interactions on the surface.     

Connecting the wetting and rheological behaviors of poly(dimethylsiloxane)-grafted silica spheres in poly(dimethylsiloxane) melts

Using dynamic light scattering, mechanical rheometry, and visual observation, the static wetting behavior of PDMS-grafted silica spheres (PDMS-g-silica) in PDMS melts is related to their rheology. A phase diagram is mapped out for a constant grafted chain length as a function of grafting density and free polymer chain length. The transition between stable and aggregated regions is determined optically and with dynamic light scattering. It is associated with a first-order wetting transition. In the stable region Newtonian behavior is observed for semidilute suspensions. The hydrodynamic brush thicknesses, deduced from viscosity measurements, correspond closely to values obtained from self-consistent field calculations for the various parameter values. At the transition, the brush collapses suddenly and shear-thinning and thixotropy appear. The rheology indicates a degree of aggregation that increases with increasing length of the free polymer, as suggested by the theory.     

Bioactive surface modification of mica and poly(dimethylsiloxane) with hydrophobins for protein immobilization

Bioactive surfaces with appropriate hydrophilicity for protein immobilization can be achieved by hydrophobin II (HFBI) self-assembly on mica and polydimethylsiloxane (PDMS) surfaces. X-ray photoelectron spectroscopy and water contact angle measurements illustrated that the surface wettability can be changed from superhydrophobic (PDMS) or superhydrophilic (mica) to moderately hydrophilic, which is suitable for protein (chicken IgG) immobilization on both substrate surfaces. The results suggest that HFBI assembly, one kind of hydrophobin from Trichoderma reesei, may be a versatile and convenient method for the immobilization of biomolecules on diverse substrates, which may have potential applications in biosensors, immunoassays, and microfluidic networks.     

Preparation and characterization of poly(methyltetradecylsiloxane) stationary phases immobilized by gamma radiation onto zirconized silica

The preparation of stationary phases with enhanced chemical stability in alkaline eluents has been the principal objective of many chromatographers. New and improved silica substrates and advanced chemical modification methods are among the possibilities being investigated to reach this objective. The present work has evaluated these two possibilities for new stationary phases. First, the silica surface was modified by reaction with zirconium tetrabutoxide to produce zirconized silica particles having about 21% (w/w) of zirconium. Then poly(methyltetradecylsiloxane) (PMTDS) was immobilized onto this surface using different doses (50-120 kGy) of gamma radiation. These new phases were characterized using elemental analysis and infrared and solid-state (29)Si-nuclear magnetic resonance (NMR) spectroscopies. These new stationary phases presented column efficiencies of about 68,000 plates m(-1), symmetric peaks for apolar compounds and retention factors that depend on the irradiation dose and show improved stability in high pH mobile phases. The separation of several pharmaceuticals at pH 11 is presented.     

Chromatographic evaluation of some stationary phases based on poly(methyloctylsiloxane) immobilized onto silica

Several stationary phases were prepared by thermal immobilization of poly(methyloctylsiloxane) onto a silica surface using different amounts of poly(methyloctylsiloaxane), and different times and temperatures of immobilization to provide different carbon contents for the phases. The chromatographic properties were evaluated using the Tanaka test. Comparison of the results obtained with literature data using hierarchical cluster analysis showed dissimilarity with most of the commercial phases. Some basic pharmaceuticals, including six benzodiazepines were separated on one of the better PMOS phases.     

聚N-异丙基丙烯酰胺硅胶键合固定相的制备与评价

以3-巯丙基三甲氧基硅烷为偶联剂,将聚N-异丙基丙烯酰胺(PNIPAM)键合到硅胶上,制得了键合固定相（SI-PNIPAM）填料,并用元素分析、红外光谱等对其进行了表征。以甲醇-水为二元流动相,用多环芳烃、碱性物质对该固定相进行了色谱评价,并考察了该固定相的适用pH范围及水解稳定性。结果表明:该固定相具有较好的色谱性能与温敏特性,并且在pH 2.5～7.5时稳定性良好。     

Preparation,evaluation, and application of a novel reversed-phase/zwitterionic/hydrophilic interaction liquid chromatographic mixed-mode stationary phase

The present study described the preparation and application of a reversed-phase/zwitterionic/hydrophilic interaction liquid chromatography stationary phase, named as SIL-PS. The SIL-PS was prepared through a four-step reaction, chemical bonding, nucleophilic addition, SN1 substitution, and sulfonation on the silica matrix. It was featured with C₁₂ alkyl chain, quaternary ammonium, tertiary amine, and sulfonate groups. After SIL-PS was packed into the stainless steel column (150?× 2.1 mm i.d.), chromatographic parameters, including acetonitrile content, pH, and ionic strength of the mobile phase, and the column temperature, were systematically investigated to study the retention mechanism. Electrostatic adsorptive/repulsive, partition, and hydrogen-bonding interactions were demonstrated to contribute to the retention. The stability of the SIL-PS was satisfactory, with relative standard deviations of retention factors of 1.93, 2.08, and 1.90% for loxoprofen, adenosine, and liquiritin, respectively. Additionally, to investigate the separation selectivity, the non-steroidal anti-inflammatory drugs, nucleobases/nucleotides, and alkaloids/glycosides were separated; the HPLC fingerprinting of the Cortex phellodendri extract was also conducted, and the separation performance was superior to that of the C18 column in terms of peak shape, resolution, and analytical time. The results revealed that the prepared SIL-PS possessed multifunctionalities for multiretention and could be promising for complicated samples.     

Nylon-6 capillary-channeled polymer fibers as a stationary phase for the mixed-mode ion exchange/reversed-phase chromatography separation of proteins

Capillary-channeled polymer (C-CP) fibers extruded from nylon-6 are used as the stationary phase for the ion-exchange/reversed-phase mixed-mode chromatographic separation of a three protein mixture. The nylon-6 C-CP fibers are packed collinearly in a 250 x 1.5-mm i.d. column with an interstitial fraction of approximately 0.6. The effects of four displacing salts at three different pHs are studied with regards to protein retention time, peak width, selectivity, and resolution for a synthetic mixture consisting of myoglobin, ribonuclease A, and lysozyme to determine the optimum mobile phase conditions. The net charge model is found to be inadequate in fully explaining the retention behavior, as the proteins are retained by anion and cation-exchange interactions, as well as hydrophobic interactions with the stationary phase. It is found that pH and displacing salt strength had a significant influence on the retention properties and resolution of the proteins.     

Hydrophobization of glass surface by adsorption of poly(dimethylsiloxane)

Silica or glass particles are introduced in a poly(dimethylsiloxane) (PDMS) matrix for various applications. A particular feature of these systems is that PDMS adsorbs on the surface of the dispersed particles, thus rendering them more hydrophobic with time. The mechanism of this process of in situ hydrophobization is still poorly understood. The major aims of the present study are (1) to quantify the rate of surface hydrophobization by PDMS and, on this basis, to discuss the mechanism of the process; (2) to compare the contact angles of surfaces that are hydrophobized by different procedures and are placed in contact with different fluid interfaces-PDMS-water, hexadecane-water, and air-water; and (3) to check how the type of surfactant affects the contact angles, viz., the effective hydrophobicity of the surface. We present experimental results for the kinetics of hydrophobization of glass surfaces, which are characterized by measuring the three-phase contact angle of glass-surfactant solution-PDMS. The data reveal two consecutive stages in the hydrophobization process: The first stage is relatively fast and the contact angle increases from 0 degrees to about 90 degrees within several minutes. This stage is explained with the physical adsorption of the PDMS chains, as a result of hydrogen-bond formation with the surface silanol groups. The second stage is much slower and hours or days are required at room temperature to reach the final contact angle (typically, 150-160 degrees). This stage is explained as grafting of the PDMS molecules on the surface by chemical reaction with the surface silanol groups. If the glass surface had been pretreated by hexamethyldisilazane (HMDS), so that CH(3) groups had blocked most of the surface silanol groups, the first stage in the hydrophobization process is almost missing-the contact angle slowly changes at room temperature from about 90 degrees up to 120 degrees. The experiments aimed to compare several hydrophobization procedures showed that PDMS ensures larger contact angle (more hydrophobic surface) than grafted alkyl chains. The contact angles at the PDMS-water and hexadecane-water interfaces were found to be very similar to each other, and much larger than that at the air-water interface. Interestingly, we found that the ionic surfactants practically do not affect the contact angle of PDMS-hydrophobized surface, whereas the nonionic surfactants reduce this angle. Similar trends are expected with silica surfaces, as well.     

Poly(2-N-carbazolylethyl acrylate)-modified silica as a new polymeric stationary phase for reversed-phase high-performance liquid chromatography

Poly(2-N-carbazolylethyl acrylate) having terminal trimethoxysilyl groups was newly synthesized by radical polymerization and immobilized onto the silica surface (Sil-CEA). The chromatographic property of Sil-CEA was examined by applying polycyclic aromatic hydrocarbons as solutes. Poly(4-vinylpyridine)-modified silica (Sil-VP) and monomeric octadecylated silica (ODS) columns were used as the reference columns. Less sensitivity to molecular hydrophobicity and enhanced molecular planarity selectivity were obtained with Sil-CEA compared to ODS. On the other hand, high retention factors for the analyzed solutes and an opposite elution order for linear and disc-shaped solutes were obtained with Sil-CEA compared with Sil-VP. In this paper, the application for separation of tocopherols was also described.     

Functionality and stability of heparin immobilized onto poly(dimethylsiloxane)

Poly(dimethylsiloxane) (PDMS) has become an attractive material when working in the field of microfluidics, mainly because of the rapid prototyping process it involves. The increased surface volume ratio in microchannels makes the interaction between sample and material surface highly important, evident when handling complex biological samples such as plasma or blood. This study demonstrates a new grade of non-covalent heparin surface that adds efficient anticoagulant property to the PDMS material. The surface modification is a simple and fast one-step process performed at neutral pH, optimal when working with closed microsystems. The heparin formed a uniform and functional coating on hydrophobic PDMS with comparatively high level of antithrombin-binding capacity. In addition, long-term studies revealed that the immobilized heparin was more or less stable in the microchannels over a time of three weeks. Recalcified plasma in contact with native PDMS showed complete coagulation after 1h, while no fibrin formation was detected in plasma incubated on heparin-coated PDMS within the same time. In conclusion, we see the heparin coating developed and evaluated in this study as a tool that greatly facilitates the use of PDMS in microfluidics dealing with plasma or blood samples.     

Liquid chromatography and characterization of ether-functionalized imidazolium ionic liquids on mixed-mode reversed-phase/cation exchange stationary phase

A new series [C_nO_mmim]Cl of imidazolium cation-based ionic liquids (ILs), with an ether functional group on the alkyl side-chain, has been prepared. The possibility of analyzing the ionic liquids by high performance liquid chromatography (HPLC) was investigated on mixed-mode reversed/cation exchange stationary phase with the aqueous-acetonitrile mobile phase. Elution parameters, such as retention factor, selectivity and column efficiency, were studied as functions of mobile phase composition and pH. The ILs were characterized by elemental analysis, and infrared, UV and ¹H, ¹³C NMR spectroscopy.     

Thermal behaviour of poly(dimethylsiloxane) hybrid silicas prepared by radiation grafting

This paper reports our investigation regarding the thermal properties of new polymer-silica hybrid materials obtained by radiation grafting. The polymer poly(dimethylsiloxane),bis(3-aminopropyl)terminated is γ-grafted on a silica gel surface. The thermal behaviour of γ-grafted hybrid materials reveals remarkable differences compared to the thermal behaviour of physically adsorbed polymers. These differences allow us to assess the ability of γ-rays to produce a polymer chemically bonded on a silica surface. The chemical bonds formed by irradiation give to the polymer a high conformational stability confirmed by DTA analysis.