RESUMEN
Coated capillaries can be advantageous in many capillary electrophoretic applications where nonaqueous background electrolytes are used. In the present work, a new dynamic polymer coating (poly(glycidylmethacrylate-co-N-vinylpyrrolidone)) for methanol-based background electrolytes is introduced. The magnitude and stability of electroosmotic flow was investigated with coated capillaries at pH* values of 3, 7.8, and 10.4 in methanol. At pH* 7.8 and 10.4 the electroosmotic flow was negligible and repeatable. On the other hand, at pH* 3 a weak, unstable electroosmotic flow was observed, due to a change in the conformation of the polymer under acidic conditions. The dynamically coated capillaries were successfully applied to the separations of cationic drugs, phenols, and benzoic acids. The synthesis and characterization of the polymer are described in detail.
Asunto(s)
Materiales Biocompatibles Revestidos , Codeína/análogos & derivados , Electroforesis Capilar/métodos , Concentración de Iones de Hidrógeno , Alprenolol/aislamiento & purificación , Materiales Biocompatibles Revestidos/química , Codeína/aislamiento & purificación , Electrólitos , Electroforesis Capilar/instrumentación , Efedrina/aislamiento & purificación , Luz , Espectroscopía de Resonancia Magnética , Metanfetamina/aislamiento & purificación , Metanol , Propranolol/aislamiento & purificación , Dispersión de RadiaciónRESUMEN
This study describes three ways to couple micellar electrokinetic chromatography (MEKC) on-line with electrospray ionization mass spectrometry (ESI-MS) for the analysis of N-methylcarbamate pesticides. The methods involved the use of a partial filling (PF) technique under basic conditions and the use of reverse migrating micelles (RMMs) under acidic and basic conditions. The use of RMMs in basic electrolyte solutions required coated capillaries with low electroosmotic flows, and capillaries coated with anionic poly(sodium 2-acrylamide-2-methylpropanesulfonate) were selected for the purpose. Before the on-line MEKC-ESI-MS coupling, the MEKC and MS conditions were separately optimized under off-line conditions. The methods were compared in terms of detection limits and the stability of the electrospray process. The PF method offered good separation but poorer stability of the electrospray relative to the other methods. A more stable electrospray performance was obtained with use of RMMs in acidic electrolyte solutions, but some of the analytes were protonated and could not be detected due to the increase in their retention factors. However, with the use of anionic polymer-coated capillaries and RMMs at pH 8.5, all analytes were successfully separated. The high-salt stacking method was applied to improve the sensitivity of MEKC-ESI-MS and the detection limits were in the range of 0.04-2.0 microg/ml.
Asunto(s)
Carbamatos/química , Cromatografía Capilar Electrocinética Micelar/métodos , Insecticidas/química , Micelas , Espectrometría de Masa por Ionización de Electrospray/métodos , Sensibilidad y EspecificidadRESUMEN
A fast and easy method was sought for determination of the iridoid glycosides catalpol, ketologanin, verbenalin, loganin, 8-epi-loganic acid, geniposidic acid and 10-cinnamoyl catalpol in plant samples. The method involved micellar electrokinetic capillary chromatography (MEKC) coupled on-line to mass spectrometry. The partial filling technique and electrospray ionization were used. Seven iridoid glycosides could be separated with use of MEKC under basic conditions. However, 8-epi-loganic acid and geniposidic acid could not be detected simultaneously with the five neutral iridoid glycosides by mass spectrometry. Therefore, only the neutral iridoid glycosides were screened from plant samples. Catalpol, verbenalin, loganin and possibly 10-cinnamoyl catalpol were found in an examination of seven plant species in the genera Plantago, Veronica, Melampyrum, Succisa, and Valeriana. Aucubin, which was not included in the sample mixture used in method development because of overlapping with catalpol in MEKC, was also detected. The limits of detection for the iridoid glycosides, both at the UV and at the mass spectrometer, are given.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar/métodos , Glucósidos/análisis , Espectrometría de Masas/métodos , Plantas Medicinales/química , Piranos/análisis , Glucósidos/química , Concentración de Iones de Hidrógeno , Glucósidos Iridoides , Glicósidos Iridoides , Iridoides , Estructura Molecular , Peso Molecular , Plantago/química , Piranos/química , Scrophulariaceae/química , Especificidad de la Especie , Espectrofotometría Ultravioleta , Valerianaceae/químicaRESUMEN
Liposomes made of mixtures of zwitterionic and anionic lipids were investigated by means of capillary electrophoresis and dynamic light scattering. The influence of the molar lipid ratio and of the buffers, used in the running electrolyte solution, on the physical characteristics of the liposomes were investigated. Data on effective electrophoretic mobilities, total charges as well as sizes of the liposomes are given. In addition, examples on the use of liposomes as carriers in electrokinetic capillary electrophoresis for the separation of benzene derivatives, steroids, and phenols are shown.
Asunto(s)
Liposomas/química , Electroforesis Capilar , Liposomas/análisisRESUMEN
Liposomes are small membrane-enclosed vesicles composed of either natural or synthetic lipids. Their size can be adjusted on a wide scale and they can be made with well-defined compositions. While liposomes have been extensively used as model biomembranes they have also gained a considerable degree of attention as carriers for drugs as well as for genetic material. The physical properties of liposomes are critically dependent on their chemical composition. In this study liposomes were applied as pseudostationary phases in electrokinetic capillary chromatography. Various negatively charged liposomes, consisting of mixtures of zwitterionic and anionic lipids, were investigated. Major emphasis was put on clarifying the effects of the total lipid concentration, the lipid molar ratio, the lipid head group, and the buffer on the capillary electrophoretic separation of neutral analytes. In addition, the influence of the physical state of the membrane, ie., gel vs. fluid, on the separation was investigated. Corticosteroids were applied as model analytes.
Asunto(s)
Corticoesteroides/análisis , Electroforesis Capilar/métodos , Liposomas , Fosfolípidos , Corticoesteroides/aislamiento & purificación , Portadores de Fármacos , Cinética , Relación Estructura-ActividadRESUMEN
Polygalacturonic acid, a linear homopolysaccharide, was investigated by capillary electrophoresis (CE) using linear polyacrylamide-coated capillaries and laser-induced fluorescence (LIF) detection. A successful separation of its fluorescently labeled oligomers was achieved through sieving in polyacrylamide entangled matrices. The reaction conditions for the derivatization of polygalacturonic acid were optimized. In studying the interactions between polygalacturonic acid and various metal ions, the end-label, free-solution electrophoretic (ELFSE) technique, developed earlier in our laboratory (Sudor, J., Novotny, M. V., Anal. Chem. 1995, 67, 4205-4209) was found preferable to the sieving method. ELFSE is fast and convenient in that no polymer solutions are needed for the separation. The investigation showed that for the moderately large oligomers, the strongest binding occurred with calcium and cadmium ions, while the smallest interaction was observed with magnesium ions.
Asunto(s)
Oligosacáridos/química , Oligosacáridos/aislamiento & purificación , Pectinas/química , Cadmio , Calcio , Cationes Bivalentes , Electroforesis Capilar/métodos , Electroforesis en Gel de Poliacrilamida/métodos , Indicadores y Reactivos , Bases de Schiff , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
The one-line combination of micellar electrokinetic chromatography (MECC) with electrospray ionization-mass spectrometry (ESI-MS) was investigated using some corticosteroids as model components. The on-line technique is difficult because the micelles in the electrolyte solution tend to soil the mass spectrometer and lower the sensitivity of the spectrometer. To prevent the micelles from reaching the mass spectrometer, several techniques have been developed among which is the partial filling (PF) technique. In this study the PF-MECC technique was investigated in an on-line MECC-ESI-MS study of mixtures of corticosteroids. Because the compounds are uncharged, partitioning or interaction with micelles is required to achieve separation. Surfactant solutions of sodium dodecyl sulfate (SDS), sodium cholate (SC), and SDS/SC mixtures were used as micellar phase. Good MECC separation was achieved after optimization of the surfactant concentrations and the length of the injected micellar plug. Both hydrodynamic and electrokinetic injections of the micellar solutions were tested. A basic ammonium acetate solution was used as the CE electrolyte solution. The ESI-MS analysis of the compounds was performed in the positive ionization mode, using an acidic sheath liquid. Because of the low MS intensities of the corticosteroids, the peaks were isolated during the MS runs. MS-MS and MS-MS-MS data on the corticosteroids were obtained by off-line injection of the compounds.
Asunto(s)
Corticoesteroides/análisis , Cromatografía/métodos , Electroforesis Capilar/métodos , Espectrometría de Masas/métodos , Electrólitos , Iones , Micelas , Estructura Molecular , SolucionesRESUMEN
In addition to high efficiency, short analysis times and small sample volumes, a further attractive feature of capillary electrophoretic techniques is the possibility to achieve, high selectivities. Usually, selectivity control also allows improvement in the resolution. A simple way to enhance the selectivity of capillary electrophoretic separations is to add one or more surfactants above their critical micelle concentration, or in the case of chiral separations to add a chiral selector to the background electrolyte. Because of the dynamic structure of micelles, the aggregation of monomers and size of the micelles can be easily adjusted. This review describes the various type of surfactants used in micellar electrokinetic capillary chromatography, and the chiral selectors employed in enantiomeric separations by capillary electrophoresis. Factors affecting the selectivity are noted. A brief discussion is included of the selectivity enhancement obtainable in non-aqueous media.
Asunto(s)
Electroforesis Capilar/métodos , Micelas , Preparaciones Farmacéuticas/química , Preparaciones Farmacéuticas/aislamiento & purificación , Dodecil Sulfato de Sodio/química , Tensoactivos/química , Tampones (Química) , Ciclodextrinas , Dextrinas , Concentración de Iones de Hidrógeno , Sensibilidad y Especificidad , Solventes/química , Estereoisomerismo , Tensoactivos/clasificación , TemperaturaRESUMEN
The application of mixed micellar electrokinetic capillary electrochromatography to the determination of the corticosteroid hormones cortisone, cortisol (hydrocortisone), and dexamethasone in serum samples is demonstrated. The serum samples were enzymatically hydrolyzed, precipitated, solid-phase extracted, and analyzed by capillary electrophoresis. A micellar system of sodium dodecyl sulfate and sodium cholate buffered with an organic compound provided the electrolyte solution. Spiked blank serum samples were used for the linearity testing and limits of detection and quantitation were measured. Patient samples were analyzed and the concentrations of cortisol and cortisone were measured. The method, which was fast and easy to perform, was confirmed to be useful for the determination of corticosteroids in serum.
Asunto(s)
Corticoesteroides/sangre , Cromatografía/métodos , Electroforesis Capilar/métodos , Ácido Cólico , Ácidos Cólicos/química , Cortisona/sangre , Dexametasona/sangre , Humanos , Hidrocortisona/sangre , Dodecil Sulfato de Sodio/química , Tensoactivos/químicaRESUMEN
Selectivity and resolution were studied for the separation of seven corticosteroids by micellar electrokinetic capillary chromatography (MEKC) using a mixed micellar solution of sodium dodecyl sulfate (SDS) and sodium cholate (SC), buffered with 3-(N-morpholino)propanesulfonic acid (MOPS) or 3-[(1,1-dimethyl-2-hydroxyethyl)amino]-2-hydroxypropane sulfonic acid (AMPSO). The changes in selectivity were compared for the AMPSO-SDS-SC system by varying the pH and the concentrations of AMPSO, SDS and SC. The experimental design started with the central composite design and continued in a sequential manner. The optimum selectivity for the separation of the corticosteroids was calculated from the analyte migration times and the analyte velocities, by using empirical quadratic regression models. Satisfactory regression fits and coefficients of determination for prediction were obtained with cross-validated models. To optimize the resolution, the physical parameters of capillary length and analysis time were varied under the conditions optimal for the selectivity. In both the selectivity and the resolution, optimization the overall optimum was determined by using the desirability function technique. Analysis times were controlled by using 1,3-diaminopropane to influence the electroosmotic flow velocity (veo). The voltage was kept constant, which resulted in higher electric field strength in shorter capillaries. No changes in the selectivity were observed when 1,3-diaminopropane was used to control the electroosmotic flow velocity. Such an optimization technique, where the chemical and physical factors affecting the separation are treated independently, seemed to be effective for finding the best possible resolution for the corticosteroids.
Asunto(s)
Corticoesteroides/aislamiento & purificación , Ácidos Cólicos , Cromatografía/métodos , Micelas , Dodecil Sulfato de Sodio , Tampones (Química) , Ácido Cólico , Cromatografía/estadística & datos numéricos , Electrólitos , Concentración de Iones de Hidrógeno , Modelos Estadísticos , Análisis de Regresión , Sensibilidad y Especificidad , SolucionesRESUMEN
The corticosteroids studied can be effectively separated by employing micellar electrokinetic capillary chromatography (MECC). The effect of pH, borate concentration and the sodium dodecyl sulfate (SDS) concentration on both the resolution and the selectivity was studied under 15 different experimental conditions. The experimental design was similar to the central composite design approach. Empirical quadratic regression models were derived for analyte migration time, band broadening and analyte velocity. Satisfactory regression fits and coefficients of determination for prediction were obtained with cross-validated models. The models for analyte migration time and analyte velocity were in good agreement with theory. Modeling of the band broadening seemed to be somewhat more complicated. Optimum conditions for resolution and selectivity were different. This is due to the fact that selectivity studies ignore the electroosmotic and band broadening properties of different electrolyte solutions. However, the study of the selectivity yielded good information about the suitability of the electrolyte systems for the particular separation problem. Although a high solubilizing power of SDS caused the corticosteroids to partition strongly into the SDS micelles, a good separation could be achieved at low SDS concentrations.