Micellar electrokinetic chromatography separation technique

The effects of pH on electrokinetic velocities in micellar electrokinetic chromatography was studied by using sodium dodecyl sulfate solutions [179]. Micellar electrokinetic capillary chromatography with a sodium dodecyl sulfate pseudostationary phase has been used to determine the partition constants for nitrophenols, thiazolylazo dyes, and metal chelate compounds [180]. A similar technique was used to separate hydroquinone and some of its ether derivatives. This analysis is suitable for the determination of hydroquinone in skin-toning creams [181]. The ingredients of antipyretic analgesic preparations have also been determined by this technique [182], The addition of sodium dodecyl sulfate improves the peak shapes and resolution in chiral separations by micellar electrokinetic chromatography [183]. 

Separation in Micellar Electrokinetic Chromatography (MEKC) is based on partitioning of the analyte molecules between the aqueous run buffer and the core of micelles, which are contained in the run buffer. The technique is essentially a hybrid between CE and liquid chromatography (LC). The run buffer and micelles are moved through the capillary by an applied electric field. The analytes are dragged with the bulk solution. Similar to LC, the analytes partition between two phases, in this case two mobile phases, the hydrophilic run buffer and the hydrophobic micelles. Unlike other electrophoresis modes, MEKC can distinguish between different neutral compounds according to their hydrophobicity. 

The capillary electrophoresis (CE) technique can be used with micellar phases. The first use of such phases with CE was presented in 1984 by Terabe, who called the technique micellar electrokinetic chromatography (MEKC) [39]. Its success was tremendous because it opened the use of CE to noncharged molecules and species. CE became an essential separation tool especially in the field of biology. Today ten applications are published in MEKC for only one in MLC. Numerous books and review articles describe the CE technique including the use of micellar phases [40-43]. A simplified survey is presented here. 

CE is a family of techniques similar to those found in conventional electrophoresis zone electrophoresis, displacement electrophoresis, isoelectric focusing (IEF), and sieving separations. Other modes of operation unique to CE include micellar electrokinetic chromatography (MEKC) and capillary electrochromatography (CEC). 

Koezuka, K., Ozaki, H., Matsubara, N., and Terabe, S. (1997). Separation and detection of closely related peptides by micellar electrokinetic chromatography coupled with electrospray ionization mass spectrometry using the partial filling technique.. Chromatogr. B 689, 3—11. 

Pluym et al. compared the use of CE to that of HPLC in chemical and pharmaceutical quality control. They stated that CE could be considered as a complementary technique to HPLC because of its large separation capacity, its simplicity, and its economical benefits. Jimidar et al. decided that CE offers high separation efficiency and can be applied as an adjunct in HPLC method validation. Mol et al. evaluated the use of micellar electrokinetic chromatography (MEKC) coupled with electrospray ionization mass spectrometry (ESI—MS) in impurity profiling of drugs, which resulted in efficient separations. 

Dedicated applications of capillary zone electrophoresis (CZE) coupled to MS are discussed, particularly in the field of drug analysis. Development of other capillary-based electrodriven separation techniques such as non-aqueous capillary electrophoresis (NACE), micellar electrokinetic chromatography (MEKC), and capillary electrochromatography (CEC) hyphenated with MS are also treated. The successful coupling of these electromigration schemes with MS detection provides an efficient and sensitive analytical tool for the separation, quantitation, and identification of numerous pharmaceutical, biological, therapeutic, and environmental compounds. 

Nitroaromatic explosives and other nitrated organic explosives are under the normal conditions neutral compounds and therefore cannot be separated directly by capillary zone electrophoresis (CZE) technique. Another separation vector must be introduced in order to achieve the resolution between the solutes. Micellar electrokinetic chromatography (MEKC) is typically employed on microchip scene for separation of nitroaromatic explosives. 

Capillary electrochromatography (CEC) is a rapidly emerging technique that adds a new dimension to current separation science. The major "news" in this method is that the hydrodynamic flow of the eluting liquid, which is typical of HPLC, is replaced by a flow driven by electro-endoosmosis. This increases considerably the selection of available separation mechanisms. For example, combinations of traditional processes such as reversed-phase- or ion-exchange- separations with electromigration techniques are now possible. Also, CEC is opening new horizons in the separation of non-polar compounds, and thus represents an alternative to the widely used micellar electrokinetic chromatography. 

Capillary electrophoresis is a general term that is used to describe a number of different separation techniques. Capillary zone electrophoresis (CZE) is the classic technique and is therefore usually referred to as just CE. Other techniques include micellar electrokinetic chromatography (MEKC), capillary isoelectric focusing, and capillary isotachophoresis. CZE and MEKC are the predominant techniques and are those used herein, so only they will be discussed in detail here. 

The FITC labeling method was also applied to chiral separations of amino acids on a microchip to determine the enantiomeric ratios of amino acids found on a meteorite [27], Since biotic amino acids are normally single enantiomers, chiral separations of amino acids are not truly clinical in nature, but illustrate the potential for chiral separations of small molecules of clinical interest. Ma-thies and co-workers used this technique to search for evidence of life in extraterrestrial environments. Enantiomeric forms of Val, Ala, Glu, and Asp could be discriminated by addition of a-, (3-, or y-cyclodextrin (CD) to the run buffer. Improved resolution with faster separations was found with respect to conventional CE. This method has been modified, by addition of SDS to the buffer, to perform cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) [28]. Increasing the SDS concentration decreased the magnitude of elec-troosmotic flow (EOF), increasing the effective migration distance, and therefore the resolution on the microchips. 

The majority of enantioseparations are performed by pressure-driven liquid chromatography. However, in the last decade other liquid-phase separation techniques have evolved and demonstrated their usefulness for enantioseparations, including supercritical fluid chromatography (SFC), capillary electrophoresis (CE), micellar electrokinetic chromatography (MEKC), and open-tubular and packed-bed electrochromatography (OT-EC and CEC). 

Micelles and cyclodextrins are the most common reagents used for this technique. Micellar electrokinetic capillary chromatography (MECC or MEKC) is generally used for the separation of small molecules [6], Sodium dodecyl sulfate at concentrations from 20 to 150 mM in conjunction with 20 mM borate buffer (pH 9.3) or phosphate buffer (pH 7.0) represent the most common operating conditions. The mechanism of separation is related to reversed-phase liquid chromatography, at least for neutral solutes. Organic solvents such as 5-20% methanol or acetonitrile are useful to modify selectivity when there is too much retention in the system. Alternative surfactants such as bile salts (sodium cholate), cationic surfactants (cetyltrimethy-lammonium bromide), nonionic surfactants (poly-oxyethylene-23-lauryl ether), and alkyl glucosides can be used as well. 

Various papers related to the simultaneous determination of creatinine and uric acid can be found in the hterature. Several authors have developed capillary zone electrophoresis (CZE) methods for simultaneous analysis of these compounds in urine. The CE analysis of these renal markers offers some advantages when compared with chromatography, such as shortened separation time, reduced reagent consumption, and increased resolution. Capillar micellar electrokinetic chromatography has been applied to the simultaneous separation of creatinine and uric acid in human plasma and urine. However, chromatographic techniques are widely accepted for the determination of these compounds. Reversed-phase and ion... 

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