Boronate functionalised polymer monoliths for microscale affinity chromatography.
Novel macroporous monolithic stationary phase materials suitable for microscale boronate affinity chromatography were developed.
Boronic Acids↗
Biomedical subjects
Emily F Hilder
Publications and source records attributed to Emily F Hilder.
Sensitive determination of carbohydrates labelled with p-nitroaniline by capillary electrophoresis with photometric detection using a 406 nm light-emitting diode.
p-Nitroaniline was explored as a derivatising reagent for UV absorbance detection of carbohydrates after separation by CE. This derivatising agent has three advantages: first, it has excellent water solubility; second, it has high molar absorptivity; and third, it is possible to obtain sensitive detection using a UV or blue light-emitting diode (LED) as the light source. The labelling reaction took less than 30 min to complete with high reaction yield. The separation process was modelled and optimised using an artificial neural network. Nine carbohydrates were separated by a CE system within 16 min using a 0.17 M boric acid buffer at pH 9.7. On-column LED detection at 406 nm allowed the detection of carbohydrates with good detection limits (<1.1 microM or 8.8 fmol) and reproducible quantification in the concentration range of 2.6-200 microM. This method was applied successfully to the determination of component carbohydrates in some food samples.
Aniline Compounds↗
Monolithic stationary phases for fast ion chromatography and capillary electrochromatography of inorganic ions.
The focus of this review is on current developments in monolithic stationary phases for the fast analysis of inorganic ions and other small molecules in ion chromatography (IC) and capillary electrochromatography (CEC), concentrating in particular on the properties of organic (polymer) monolithic materials in comparison to inorganic (silica-based) monoliths. The applicability of these materials for fast IC is discussed in the context of recent publications, including the range of synthesis and modification procedures described. While commercial monolithic silica columns already show promising results on current IC instrumentation, polymer-based monolithic stationary phases are currently predominantly used in the capillary format on modified micro-IC systems. However, they are beginning to find application in IC particularly under high pH conditions, with the potential to replace their particle-packed counterparts.
Chromatography, Liquid↗
Towards high capacity latex-coated porous polymer monoliths as ion-exchange stationary phases.
The preparation of high capacity agglomerated monolithic ion-exchangers for capillary ion chromatography is described. Post-modification of reactive monoliths was investigated as an alternative to co-polymerisation of a suitable functional monomer with an overarching goal of increasing ion-exchange capacity. Direct sulfonation of poly styrene-co-divinyl benzene monoliths using concentrated sulfuric acid or chlorosulfonic acid was unsuccessful even for monoliths containing as low as 8% crosslinker. In contrast, chemical transformation of reactive monoliths containing glycidyl methacrylate was used to increase the ion-exchange capacity by up to more than thirty-fold with ion exchange capacities of 14-29 microequiv g(-1) achieved. Three different reactions were considered, including reaction with 4-hydroxybenenesulfonic acid under basic conditions; reaction with thiobenzoic acid followed by transformation to a reactive thiol and the subsequent oxidation to the sulfonic acid; and direct sulfonation with sodium sulfite. Of these, the reaction with sodium sulfite resulted in the most significant increase in the capacity and the best separation performance. In the isocratic mode separation efficiencies of over 13,500 plates m(-1) were observed (for iodate). The separation of seven inorganic anions was also demonstrated using a hydroxide gradient.
Chromatography, Ion Exchange↗
Preparation and characterisation of anion-exchange latex-coated silica monoliths for capillary electrochromatography.
Silica monoliths coated with functionalised latex particles have been prepared for use in monolithic ion-exchange capillary electrochromatography (IE-CEC) for the separation of inorganic anions. The ion-exchange monoliths were prepared using 70 nm quaternary ammonium, anion-exchange latex particles, which were bound electrostatically to a monolithic silica skeleton synthesised in a fused silica capillary. The resulting stationary phases were characterised in terms of their chromatographic performance and capacity. The capacity of a 50 microm diameter 25 cm latex-coated silica monolith was found to be 0.342 nanoequivalents and 80,000 theoretical plates per column were typically achieved for weakly retained anions, with lower efficiency being observed for analytes exhibiting strong ion-exchange interaction with the stationary phase. The electroosmotic flow (EOF) was reversed after the latex-coating was applied (-25.96 m2 V(-1) s(-1), relative standard deviation (RSD) 2.8%) and resulted in anions being separated in the co-EOF mode. Ion-exchange interactions between the analytes and the stationary phase were manipulated by varying the ion-exchange selectivity coefficient and the concentration of a competing ion (phosphate or perchlorate) present in the electrolyte. Large concentrations of competing ion (greater than 1M phosphate or 200 mM perchlorate) were required to completely suppress ion-exchange interactions, which highlighted the significant retention effects that could be achieved using monolithic columns compared to open tubular columns, without the problems associated with particle-packed columns. The latex-coated silica monoliths were easily produced in bulk quantities and performed reproducibly in acidic electrolytes. The high permeability and beneficial phase ratio makes these columns ideal for micro-LC and preconcentration applications.
Chromatography, Ion Exchange↗
Latex-functionalized monolithic columns for the separation of carbohydrates by micro anion-exchange chromatography.
A novel stationary phase for micro ion chromatography has been prepared by coating a porous poly(butyl methacrylate-co-ethylene dimethacrylate-co-2-acrylamido-2-methyl-1-propanesulfonic acid) monolith with quaternary amine-functionalized latex particles via simple electrostatic binding. This stationary phase enabled the separation of saccharides in the mobile phase with a high-pH value consisting of aqueous ammonia solution in anion-exchange mode using evaporative light scattering for detection. Effects of both porous properties of the monolithic stationary phase and chromatographic conditions on the separation ability were studied. Under optimized conditions, an efficient separation of seven saccharides was achieved in less than 10 min. The stationary phase also enables the separation of saccharides obtained by the enzymatic hydrolysis of corn starch.
Anion Exchange Resins↗
Development and application of polymeric monolithic stationary phases for capillary electrochromatography.
Monolithic columns for capillary electrochromatography are receiving quite remarkable attention. This review summarizes results excerpted from numerous papers concerning this rapidly growing area with a focus on monoliths prepared from synthetic polymers. Both the simplicity of the in situ preparation and the large number of readily available chemistries make the monolithic separation media a vital alternative to capillary columns packed with particulate materials. Therefore, they are now a well-established stationary phase format in the field of capillary electrochromatography. A wide variety of synthetic approaches as well as materials used for the preparation of the monolithic stationary phases are presented in detail. The analytical potential of these columns is demonstrated with separations involving various families of compounds and different chromatographic modes.
Electrophoresis, Capillary↗
Shielded stationary phases based on porous polymer monoliths for the capillary electrochromatography of highly basic biomolecules.
A novel stationary phase for capillary electrochromatography has been prepared via photoinitiated grafting of two layers of polymer chains onto the pore surface of a porous polymer monolith. To achieve the desired retention, the original monolith with optimized porous properties was grafted with an "interior" layer consisting of the ionizable monomer, 2-acrylamido-2-methyl-1-propanesulfonic acid, followed by a "covering" layer of hydrophobic polymer chains. This technique affords monolithic CEC columns that facilitate electroosmotic flow (EOF) while preventing ionized analytes from interacting with the charged surface functionalities. Grafting of the second layer does not adversely affect the EOF. Grafting times of 30 and 60 s for AMPS and butyl acrylate, respectively, enabled the preparation of a monolith with full shielding of the analytes from the ionizable functionalities and excellent chromatographic performance. This approach allows for the first time the independent optimization of both electroosmotic flow and retention properties in CEC columns. The efficient isocratic separations of mixtures of peptides, including some that are highly basic and would be affected by unshielded charges, were routinely achieved in 40-90 s using a simple MS compatible mobile phase consisting of 20 mmol/L ammonium acetate in a 1:1 water-acetonitrile mixture.
Acrylamides↗
Poly(tetrafluoroethylene) separation capillaries for capillary electrophoresis. Properties and applications.
Poly(tetrafluoroethylene) (PTFE) is a material widely known for its inertness and excellent electrical properties. It is also transparent in the UV region and has a reasonable thermal conductivity. These properties make PTFE a suitable material for the separation capillary in capillary electrophoresis. Differences in the chemistry of the capillary wall compared to fused silica (FS) can make PTFE an interesting alternative to FS for some special applications. In this work, properties of a commercial PTFE capillary of approx. 100 microm i.d. were investigated, including the dependence of electroosmotic flow (EOF) on pH for unmodified and dynamically modified PTFE, optical properties, and practical aspects of use. The main problems encountered for the particular PTFE capillary used in this study were that it was mechanically too soft for routine usage and the crystallinity of the PTFE caused light scattering, leading to high background absorbance values in the low UV region. The profile of the EOF versus pH for bare PTFE surprisingly showed significantly negative EOF values at pH < 4.2, with an EOF of -30 x 10(-9) m2 V(-1) s(-1) being observed at pH 2.5. This is likely to be caused by either impurities or additives of basic character in the PTFE, so that after their protonation at acidic pH they establish a positive charge on the capillary wall and create a negative EOF. A stable cationic semi-permanent coating of poly(diallyldimethylammonium chloride) (PDDAC) could be established on the PTFE capillary and led to very similar magnitudes of EOF to those observed with FS. A hexadecanesulfonate coating produced a cathodic EOF of extremely high magnitude ranging between +90 and +110 x 10(-9) m2 s(-1) V(-1), which are values high enough to allow counter-EOF separation of high mobility inorganic anions. In addition, pH-independent micellar electrokinetic capillary chromatography (MEKC) separations could be easily realised due to hydrophobic adsorption of sodium dodecylsulfate (used to form the micelles) on the wall of the PTFE capillary. The use of polymers that would be mechanically more robust and optically transparent in the low-UV region should make such CE capillaries an interesting alternative to fused silica.
Electrodes↗
Porous polymer monolith for surface-enhanced laser desorption/ionization time-of-flight mass spectrometry of small molecules.
Porous poly(butyl methacrylate-co-ethylene dimethacrylate), poly(benzyl methacrylate-co-ethylene dimethacrylate), and poly(styrene-co-divinylbenzene) monoliths have been prepared on the top of standard sample plates used for matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and the modified plates were used for laser desorption/ionization mass spectrometry (LDI-MS). The hydrophobic porous surface of these monoliths enables the transfer of sufficient energy to the analyte to induce desorption and ionization prior to TOFMS analysis. Both UV and thermally initiated polymerization using a mask or circular openings in a thin gasket have been used to define spot locations matching those of the MALDI plates. The desorption/ionization ability of the monolithic materials depends on the applied laser power, the solvent used for sample preparation, and the pore size of the monoliths. The monolithic matrices are very stable and can be used even after long storage times in a typical laboratory environment without observing any deterioration of their properties. The performance of the monolithic material is demonstrated with the mass analysis of several small molecules including drugs, explosives, and acid labile compounds. The macroporous spots also enable the archiving of samples.
Acids↗
Fabrication of porous polymer monoliths covalently attached to the walls of channels in plastic microdevices.
UV-initiated grafting of plastic tubes and microfluidic chips with ethylene diacrylate followed by the preparation of porous polymer monoliths has been studied. The first step affords a thin grafted layer of polymer with a multiplicity of pendent double bonds that are then used in the second step for covalent attachment of the monolith to the wall. As clearly seen on scanning electron micrographs, this procedure prevents the formation of voids at the monolith-channel interface a problem that has always plagued approaches involving bulk polymerization in nontreated channels due to the shrinkage of the monolith during the polymerization process and its lack of compatibility with the material of the device. Irradiation with UV light through a photomask allows precise patterning specifying both the area subjected to surface modification and the location of the monolith within specific areas of the device.
Microfluidics↗
Use of ionic polymers as stationary and pseudo-stationary phases in the separation of ions by capillary electrophoresis and capillary electrochromatography.
One of the problems with capillary electrophoresis is a lack of versatility regarding manipulation of the separation selectivity. A new and potentially universal concept is to introduce an ion-exchange component into a separation so that the migration of analyte ions is influenced by both their electrophoretic mobilities and their chromatographic properties. This may be accomplished by use of capillaries filled with or coated with solid ion-exchange polymers, or by addition of a soluble ionic polymer to the background electrolyte to create a pseudo-stationary phase. While each of these methods achieves the same result, they are not competitive, but rather complementary as the problems associated by one approach are overcome by the others. Recent highlights in the field are used to illustrate the flexibility that this approach provides to electrophoretic separation of ions.
Chromatography, Micellar Electrokinetic Capillary↗
Comparison of aqueous and nonaqueous carrier electrolytes for the separation of penicillin V and related substances by capillary electrophoresis with UV and mass spectrometric detection.
A method for the determination of penicillin V together with its impurities and by-products formed during biosynthesis, using capillary electrophoresis (CE) with UV and electrospray-mass spectrometric (ESI-MS) detection is presented. Aqueous and nonaqueous electrolytes containing 20 mM ammonium acetate were investigated to determine their suitability for the separation of these analytes. These carrier electrolytes were optimized with respect to the pH and the solvent/s used (water, methanol, acetonitrile, ethanol and isopropanol) and it was shown that although the nonaqueous electrolytes offered unique separation selectivities, the best results in terms of selectivity and sensitivity were obtained for the aqueous system. Finally, the applicability of this method for the analysis of a mixture representative of a real fermentation broth was demonstrated using an aqueous carrier electrolyte with both UV and ESI-MS detection.
Acetates↗
Development and optimization of an analytical method for the determination of UV filters in suntan lotions based on microemulsion electrokinetic chromatography.
Microemulsion electrokinetic chromatography (MEEKC) has been applied to the separation of some UV filters (Eusolex 4360, Eusolex 6300, Eusolex OCR, Eusolex 2292, Eusolex 6007, Eusolex 9020, Eusolex HMS, Eusolex OS, and Eusolex 232) commonly found in suntan lotions. The composition of the microemulsion employed was optimized with respect to the best possible separation of the selected analytes using artificial neural networks (ANNs). Two parameters namely the composition of the mixed surfactant system comprising the anionic sodium dodecyl sulfate (SDS) and neutral Brij 35 and the amount of organic modifier (2-propanol) present in the aqueous phase of the microemulsion were modeled. Using an optimized MEEKC buffer consisting of 2.25 g SDS, 0.75 g Brij 35, 6.6 g 1-butanol, 0.8 g n-octane, 17.5 g 2-propanol, and 72.1 g of 10 mM borate buffer (pH 9.2), eight target analytes could be separated in under 25 min employing a diode-array detector to segregate the overlapping signals obtained for Eusolex 9020 and Eusolex HMS. Detection limits from 0.8 to 6.0 nug/mL were obtained and the calibration plots were linear over at least one order of magnitude. The optimized method could be applied to the determination of Eusolex 6300 and Eusolex 9020 in a commercial suntan lotion.
Chromatography, Micellar Electrokinetic Capillary↗
Polymeric monolithic stationary phases for capillary electrochromatography.
This review summarizes the contributions of a number of groups working in the rapidly growing area of monolithic columns for capillary electrochromatography (CEC), with a focus on those prepared from synthetic polymers. Monoliths have quickly become a well-established stationary phase format in the field of CEC. The simplicity of their in situ preparation method as well as the good control over their porous properties and surface chemistries make the monolithic separation media an attractive alternative to capillary columns packed with particulate materials. A wide variety of approaches as well as materials used for the preparation of the monolithic stationary phases are detailed. Their excellent chromatographic performance is demonstrated by numerous separations of different analytes.
Chromatography↗