Use of a partial filling technique and reverse migrating micelles in the study of N-methylcarbamate pesticides by micellar electrokinetic chromatography-electrospray ionization mass spectrometry

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.     

Chiral capillary electrophoresis-mass spectrometry: modes and applications

A review is presented to highlight several approaches for coupling capillary electrophoresis (CE) and electrospray ionization-mass spectrometry (ESI-MS) for analysis of chiral compounds. A short discussion of commercially available CE-MS instruments and interface design is followed by a detail review on various modes of chiral CE-MS. In general, for each CE-MS mode, the capabilities, applications and limitations for chiral analysis have been pointed out. The first mode, chiral capillary zone electrophoresis-mass spectrometry (CZE-MS) in which neutral derivatized cyclodextrins (CDs) are used is possible using either column coupling with voltage switching or a partial-filling technique (PFT). However, some applications of direct coupling of CZE-MS mode are also reported. The second mode is a chiral electrokinetic chromatography-mass spectrometry (EKC-MS) in which a charged chiral selector such as a sulfated beta-CD or a vancomycin could be conveniently employed. This is because these chiral selectors have a significantly higher countercurrent electrophoretic mobility which prevents the entrance of these selectors into the mass spectrometer. The combination of counter-migration and PFT demonstrates that this synergism could be successfully applied to chiral analysis of a broader range of compounds. It is well-known that the on-line coupling of micellar electrokinetic chromatography to mass spectrometry (MEKC-MS) is problematic because the high surface activity and nonvolatile nature of conventional surfactant molecules lower the electrospray ionization efficiency. However, a recent report demonstrates that this hyphenation is now possible with the use of molecular micelles. Various MEKC-ESI-MS parameters that can be used to optimize both chiral resolution and ESI response are discussed. Finally, two recent examples that demonstrate the feasibility of using either open-tubular or packed chiral CEC with MS are reviewed. This survey will attempt to cover the state-of-the-art on various modes of CE-MS from 1998 up to 2002.     

Enantiomer separation of drugs by micellar electrokinetic chromatography using chiral surfactants

A review surveying enantiomer separations by micellar electrokinetic chromatography (MEKC) using chiral surfactants is described. MEKC is one of the most popular techniques in capillary electrophoresis, where neutral compounds can be analyzed as well as charged ones, and the use of chiral micelles enable one to achieve the enantioseparation. The chiral MEKC systems are briefly reviewed according to the types of chiral surfactants along with typical applications. As chiral micelles or pseudostationary phases in MEKC, various natural and synthetic chiral surfactants are used, including several low-molecular-mass surfactants and polymerized surfactants or high-molecular-mass surfactants. Cyclodextrin modified MEKC using chiral micelles is also considered.     

On-line capillary electrophoresis-mass spectrometry using dopant-assisted atmospheric pressure photoionization: setup and system performance

The on-line coupling of capillary electrophoresis (CE) and mass spectrometry (MS) via atmospheric pressure photoionization (APPI) is demonstrated. To achieve CE-APPI-MS, an adapted coaxial sheath-flow interface was combined with an ion-trap mass spectrometer equipped with an APPI source originally designed for liquid chromatography-MS. Effective photoionization of test compounds was accomplished after optimization of several interface and MS parameters, and of the composition and flow rate of the sheath liquid. Further enhancement of the ionization efficiency could be achieved by adding a dopant, such as acetone or toluene, to the sheath liquid to aid indirect ionization. Acetone significantly increased the ionization of the polar test compounds by proton transfer, while toluene was more useful for the enhanced formation of molecular ions from nonpolar compounds. The effect of several common CE background electrolytes (BGEs) on the APPI-MS response of the analytes was also studied. It appeared that in contrast with electrospray ionization, nonvolatile BGEs do not cause suppression of analyte signals using APPI. Therefore, in CE-APPI-MS, a variety of buffers can be chosen, which obviously is a great advantage during method development. Remarkably, also sodium dodecyl sulfate (SDS) did not affect the photoionization of the test compounds, indicating a strong potential of APPI for the on-line coupling of micellar electrokinetic chromatography (MEKC) and MS.     

Highly-sensitive micellar electrokinetic chromatographic analysis of dioxin-related compounds using on-line concentration.

An application study of an on-line concentration technique of neutral analytes for micellar electrokinetic chromatography (MEKC) was carried out in environmental analysis to enhance the UV detection sensitivity. Several dioxins and related compounds, such as dibenzofuran, dibenzo-p-dioxin, 2,3- and 2,7-dichlorodibenzo-p-dioxins, and 2,3,7-trichlorodibenzo-p-dioxin, were used as test solutes. For a highly sensitive separation and detection, cyclodextrin-modified MEKC (CD-MEKC) under acidic conditions was employed as a separation mode and stacking using reverse migrating micelles and a water plug (SRW) as an on-line concentration technique. Almost a 200-fold gain in detection sensitivity was obtained for the model compounds in SRW-CD-MEKC compared to that in normal CD-MEKC without on-line concentration and the limit of detection was found to be around 0.1 ppm for each solute.     

Partial-filling micellar electrokinetic chromatography and non-aqueous capillary electrophoresis for the analysis of selected agrochemicals

Selected agrochemicals (s-triazines and phenoxy acids) have been investigated with partial-filling micellar electrokinetic chromatography (PFMEKC) and non-aqueous capillary electrophoresis (NACE). Because these two techniques are compatible for coupling of capillary electrophoresis with mass spectrometry, different conditions affecting the separation efficiency (reproducibility, method linearity) were systematically tested, and the results were compared with those from classical MEKC. The conditions tested included buffer molarity, pH, the concentrations of the organic modifier and surfactant, the applied voltage, the injection time of the sample, and the length of the partial-filling plug. The respective limits of detection (LOD) using UV-detection were determined. Reduction of the electrophoretic raw data using the mobility scale transformation (micro-scale) improved qualitative comparison of the electropherograms and the reproducibility of quantitative data (integrated peak area) thus extending this data treatment from CZE to other endoosmotic flow-driven CE-techniques such as PFMEKC and NACE.     

Comparison of electrospray ionization and atmospheric pressure photoionization for coupling of micellar electrokinetic chromatography with ion trap mass spectrometry

The performance of dopant-assisted atmospheric pressure photoionization (DA-APPI) and electrospray ionization (ESI) for the coupling of micellar electrokinetic chromatography (MEKC) with ion trap mass spectrometry (ITMS) was compared using a set of test drugs comprising basic amines, steroids, esters, phenones and a quaternary ammonium compound. The influence of the surfactant sodium dodecyl sulfate (SDS) on analyte signals was studied by infusion of sample through the CE capillary into the respective ion sources. It was found that background electrolytes (BGEs) containing 20-50mM SDS in 10mM sodium phosphate (pH 7.5) caused major ionization suppression for both polar and apolar compounds in ESI-MS, whereas APPI-MS signal intensities remained largely unaffected. ESI gave rise to the formation of SDS clusters, which occasionally may cause space-charge effects in the ion trap. Furthermore, extensive sodium-adduct formation was observed for medium polar compounds with ESI-MS, whereas these compounds were detected as their protonated molecules with APPI-MS. Using the BGE containing 20mM SDS, MEKC-ESI-MS still provides slightly lower limits of detection (LODs) (2.6-3.1muM) than MEKC-APPI-MS (4.3-6.4muM) for basic amines. For less polar compounds, highest S/Ns were obtained with APPI-MS detection (LODs, 4.5-71muM). For BGEs containing 50mM SDS, the limits of detection for MEKC-APPI-MS were more favorable (factor 1.5-12) than MEKC-ESI-MS for nearly all tested drugs. Spray shield contamination by SDS was lower in DA-APPI-MS than in ESI-MS. It is concluded that DA-APPI shows the most favorable characteristics for MEKC-MS, especially when compounds of low polarity have to be analyzed.     

On-line micellar electrokinetic chromatography-mass spectrometry: feasibility of direct introduction of non-volatile buffer and surfactant into the electrospray interface

An on-line method for the coupling of micellar electrokinetic chromatography (MEKC) and mass spectrometry (MS) is presented which allows conventional MEKC conditions to be employed without further modification. The MEKC system is coupled directly to electrospray ionization (ESI) MS using a triaxial interface. A systematic study of the influence of the surfactant concentration, the nature and concentration of buffer salts and presence of organic modifier on the interface performance indicated the feasibility of the MEKC–MS approach. Effective interfacing of MEKC was achieved with both single quadrupole and ion-trap MS instruments. Using a background electrolyte containing 20 mM sodium dodecyl sulfate (SDS) and 10 mM sodium phosphate buffer, it is demonstrated that full MEKC runs of test mixtures of mebeverine and related compounds can be monitored by ESI-MS with satisfactory sensitivity. Sub-μg/ml levels of the analytes can still be detected in full scan mode, while detection limits are in the 10–50 ng/ml range when selected ion monitoring is applied. It is shown that such sensitivity would allow full-scan MS detection of 0.1% (w/w) levels of potential impurities in mebeverine. With the ion-trap instrument successful MEKC–MS/MS experiments were carried out providing information-rich MS spectra of the related compounds. Repeated MEKC–MS analyses proved that in the course of 1 day the migration time of mebeverine remained fairly constant while the MS-signal intensity only gradually decreased to approximately 65% of its original value. Once-a-day cleaning of the first part of the ion source, which takes only 5 min, suffices to preserve an optimal interface performance for a prolonged period of time.     

A comparative study of interfaces for microchip micellar electrokinetic chromatography-electrospray ionization mass spectrometry using the surfactant ammonium dodecyl sulfate

Using ammonium dodecyl sulfate (ADS) as the surfactant, the response of three common interfaces in the direct coupling of microchip micellar electrokinetic chromatography with electrospray ionization mass spectrometry was studied. In the range of 10-40 mM surfactant, a conventional sheath liquid interface provided poorer sensitivity than both sheathless interface and low-sheath-flow interface. At a surfactant concentration <20 mM, a low-sheath-flow interface exhibited less sensitivity than a sheathless interface; however, it outperformed the sheathless interface above a concentration of 20 mM. At a surfactant concentration above 20 mM, signal reduction due to dilution of the analyte compensated by signal enhancement gained from a reduction in ion suppression effect. The difference in responses of the interfaces was mainly due to the dilution effect, whereas the effect of flow rate became an important factor when the difference in responses between the interfaces was not significant. The utility of the PMMA microchip MEKC/MS using a low-sheath-flow interface was demonstrated by the analysis of sulfonamides at a concentration of 40 mM. The interday precision was in the range of 4.9-14.5%, and the LOD was in the range of 0.34-1.03 ng/mL (MEKC/MS/MS).     

Small-angle neutron scattering of mixed ionic perfluoropolyether micellar solutions

Aqueous mixed micellar solutions of perfluoropolyether carboxylic salts with ammonium counterions have been studied by small-angle neutron scattering. Two surfactants differing in the tail length were mixed in proportions n2/n3 = 60/40 w/w, where n2 and n3 are the surfactants with two and three perfluoroisopropoxy units in the tail, respectively. The tails are chlorine-terminated. The mixed micellar solutions, in the concentration range 0.1-0.2 M and thermal interval 20-40 degrees C, show structural characteristics of the interfacial shell that are very similar to ammonium n2 micellar solutions previously investigated; thus, the physics of the interfacial region is dominated by the polar head and counterion. The shape and dimensions of the micelles are influenced by the presence of the n3 surfactant, whose chain length in the micelle is 2 A longer than that of the n2 surfactant. The n3 surfactant favors the ellipsoidal shape in the concentration range 0.1-0.2 M with a 1/2 ionization degree of n2 micelles. The very low surface charge of the mixed micelles is attributed to the increase in hydrophobic interactions between the surfactant tails, due to the longer n3 surfactant molecules in micelles. The closer packing of the tails decreases the micellar curvature and the repulsions between the polar heads, by surface charge neutralization of counterions migrating from the Gouy-Chapman diffuse layer, leading to micellar growth in ellipsoids with greater axial ratios.     

Capillary electrophoresis-mass spectrometry: recent advances to the analysis of small achiral and chiral solutes

We describe here the state-of-the-art development of on-line capillary electrophoresis-mass spectrometry (CE-MS) over the last two years. Technological developments included are novel designs of new interfaces and ionization sources, new capillary coatings, buffers, and micelles as well as application of various modes of CE-MS published in the recent literature. The areas of CE-MS application in analysis of small achiral and chiral solutes are covered in sections that highlight the recent advances and possibilities of each mode of CE-MS. Application areas reviewed in this paper include achiral and chiral pharmaceuticals, agrochemicals, carbohydrates, and small peptides. The separation of enantiomers using micellar electrokinetic chromatography (MEKC)-MS with molecular micelles and capillary electrochromatography (CEC)-MS using pack tapered columns appears to provide good tolerance to electrospray stability for routine on-line CE-MS. These two modes seem to be very suitable for sensitive detection of chiral pharmaceuticals in biological samples, but their use will probably increase in the near future. Overall, it seems that one mode of CE-MS, in particular capillary zone electrophoresis (CZE)-MS, is now recognized as established technique for analysis of small charged solutes, but other modes, such as MEKC-MS and CEC-MS, are still within a period of development in terms of both MS-compatible pseudostationary phases and columns as well as applications.     

Micellar electrokinetic chromatography: current developments and future

This review highlights recent methodological and instrumental advances in micellar electrokinetic chromatography (MEKC). Enhancements in sensitivity and selectivity of the technique through the use of on-line preconcentration approaches (stacking and sweeping) and nonconventional pseudostationary phases, namely nonionic and zwitterionic surfactants, mixed micelles and polymers, are discussed in detail. Laser-induced fluorescence and mass spectrometry, as alternatives to UV-absorption detection, have been covered to evaluate their advantages and limitations when applied to analysis in an MEKC format. Some thoughts on future directions in this area such as in-capillary reactions, coated capillaries and MEKC on microchips are also presented.     

Direct coupling of micellar electrokinetic chromatography to mass spectrometry using a volatile buffer system based on perfluorooctanoic acid and ammonia

Direct coupling of micellar electrokinetic chromatography (MEKC) to mass spectrometry (MS) without employing partial filling is considered to be a challenge. One way of solving the problem would be the use of an MS-compatible surfactant. In the present study, the applicability of a series of surfactants (sodium dodecyl sulfate (SDS), lauric acid, cholic acid and perfluorated carboxylic acids) have been investigated both in terms of separation performance and MS compatibility. It was found that a MEKC system based on perfluorooctanoic acid (PFOA) and ammonia gave excellent results. The separation performance of the suggested system is comparable to the one obtained with standard systems based on SDS and sodium borate buffer although the selectivity is different. The electrospray ionization MS signal of the analytes is not seriously suppressed even at a PFOA concentration of 100 mM. Clusters are formed but their intensities are relatively low and comparable to those obtained with acetic acid. PFOA is volatile enough to allow long-term use, 30 h of continuous use has been recorded without any signs of decreasing performance. After use residual PFOA is easily removed from the ion-source (no memory effects). Furthermore, quantitation of trace impurities is possible at 25 ppb level when employing selected ion monitoring.     

MEKC: an update focusing on practical aspects

This paper reviews recent methodological and instrumental advances in MEKC. Improvements in sensitivity arising from the use of on-line sample concentration (sweeping, stacking, and combination of both protocols) and derivatization (in-capillary reactions and coupling with flow-injection systems) and improvements in resolution obtained by changing the composition of the BGE (e.g., with organic modifiers, ionic liquids, nonionic and zwitterionic surfactants, mixed micelles, and vesicles) or using coated capillaries are discussed in detail. In addition, MS and LIF spectroscopy are examined in relation to their advantages and restrictions as applied to MEKC analysis. Some thoughts on potential future directions are also expressed.     

On-line capillary electrophoresis-electrospray mass spectrometry for the stereoselective analysis of drugs and metabolites

The on-line combination of partial-filling capillary electrophoresis and electrospray ionization mass spectrometry was demonstrated for the enantioseparation of pharmaceutical drugs and metabolites, namely amphetamines, methadone, venlafaxine and selected tropane alkaloids. The partial-filling technique proved to be a suitable and efficient approach to avoid mass spectrometry (MS) source contamination, as well as signal suppression due to nonvolatile additives. To achieve chiral separation, various chiral selectors were applied, including neutral and particularly negatively charged cyclodextrins. Because of the countercurrent contribution, charged cyclodextrins were found more suitable for the on-line MS detection of separated enantiomers. Hyphenation of capillary electrophoresis (CE) with mass spectrometry was found appropriate for the stereoselective analysis of methadone in real serum samples. Moreover, the use of MS in the selected ion monitoring mode resulted in a very high selectivity, as well as improved sensitivity compared to UV detection. Finally, with atropine as a model compound, the quantitative performances of the method were evaluated and showed high sensitivity, as well as good repeatability in terms of migration time and peak area ratio.     

Widening of the elution window in micellar electrokinetic chromatography with cationic surfactants. II. Cationic additives and modifiers of the electroosmotic flow.

In micellar electrokinetic chromatography (MEKC) with cationic surfactants the migration window is significantly narrower than with anionic surfactants. In order to overcome this disadvantage of cationic surfactants, it is investigated whether it is possible to widen the migration window by reducing the velocity of the aqueous phase while the electrophoretic mobility of the micelles is maintained. Short chain alkylammonium compounds, hexamethonium bromide and hydroxypropylmethylcellulose are tested as additives to the separation electrolyte with the potential to improve the migration window via reducing the velocity of the electroosmotic flow. It will be shown that these modifiers can be successfully used in order to widen the migration window in MEKC with cationic surfactant employing an alkyltrimethylammonium bromide as micelle forming agents. Influence of the modifiers selected on retention of neutral and acidic solutes and on efficiency of the separation system is investigated.     

Online transient micellar phase concentration of anions using CTAB in CE

A transient micellar phase extractor using CTAB was described for the online sample concentration of various anionic analytes (drugs and herbicides) in CE. Stacking and separation was performed at neutral pH in coelectroosmotic flow in a hexadimethrine bromide coated fused‐silica capillary. A micellar plug (e.g. 10 mM CTAB) was injected prior to hydrodynamic injection of the analytes prepared in aqueous organic solvent (e.g. with 30% ACN). In the presence of an electric field, the micelles interacted with the anions inside the capillary. This was followed by selective analyte focusing via the mechanism of micelle to solvent stacking. The micelles acted as transient extractor because the stacking ends when the injected micelles completely migrated through the boundary between the sample and micellar plug. Fundamental studies were performed (effect of surfactant concentration, etc.) and the technique yielded 13‐ to 30‐fold improvements in peak height. A stacking CE method in conjunction with liquid–liquid extraction was also tested for the detection of the herbicides fenoprop and mecoprop in fortified drinking water at analyte concentration levels relevant to Australian Drinking Water Guidelines.     

Solute-solvent interactions in micellar electrokinetic chromatography: V. Factors that produce peak splitting

The experimental conditions that produce analyte peak splitting in micellar electrokinetic capillary chromatography (MEKC) have been systematically investigated. The system studied was a neutral phosphate buffer and sodium dodecyl sulfate (SDS) micelles as pseudostationary phase. A number of analytes showing a wide variety of hydrophobicity values and several organic solvents as sample diluents have been tested. Peak splitting phenomena are mainly due to the presence of organic solvent in the sample solution. They increase with the hydrophobicity of the analyte and decrease with the increase of the surfactant concentration. When hydrophobic compounds are analyzed the suggested ways to avoid split peaks are: (i) the use of 1-propanol or 1-butanol as sample diluent instead of methanol or acetonitrile or (ii) the use of high concentration of surfactant in the separating solution when the analyte must be dissolved in pure methanol or acetonitrile.     

Recent advances in micellar electrokinetic chromatography

This review contains nearly 200 reference citations, and covers advances in electrokinetic capillary chromatography based on micelles, including stabilized micelle complexes, polymeric and mixed micelles from 2003-2004. Detection strategies, analyte determinations, and applications in micellar electrokinetic capillary chromatography (MEKC) are discussed. Information regarding methods of analyte concentration, analyte specific analyses, and nonstandard micelles has been summarized in tabular form to provide a means of rapid access to information pertinent to the reader.     

On-line concentration by field-enhanced sample injection with reverse migrating micelles in micellar electrokinetic capillary chromatography for the analysis of flavonoids in Epimedium brevicornum Maxim

A simple and sensitive micellar electrokinetic capillary chromatography (MEKC) method was developed for the separation and determination of six flavonoids in Epimedium brevicornum Maxim. Field-enhanced sample injection with reverse migrating micelles (FESI-RMM) was used for on-line concentration of the flavonoids. An electrolyte containing 20 mM H3PO4, 100 mM SDS, 20% acetonitrile and 2% 2-propanol (pH 2.0) was chosen as the electrophoretic buffer. By optimizing the stacking conditions, about 40-360-fold improvement in the detection sensitivity was obtained for the flavonoids.