Capillary electrophoresis chiral resolution

Chromatography and Capillary Electrophoresis Chromatographic resolution is oriented toward the exclusive separation of chiral substances using columns with a chiral stationary phase from which the enantiomers are separated through diastereomeric interactions. Resolution of enantiomers contained in the mobile phase (gas or liquid)... 

Capillary Electrophoresis. Capillary electrophoresis (ce) or capillary 2one electrophoresis (c2e), a relatively recent addition to the arsenal of analytical techniques (20,21), has also been demonstrated as a powerful chiral separation method. Its high resolution capabiUty and lower sample loading relative to hplc makes it ideal for the separation of minute amounts of components in complex biological mixtures (22,23). 

Enantioresolution in capillary electrophoresis (CE) is typically achieved with the help of chiral additives dissolved in the background electrolyte. A number of low as well as high molecular weight compounds such as proteins, antibiotics, crown ethers, and cyclodextrins have already been tested and optimized. Since the mechanism of retention and resolution remains ambiguous, the selection of an additive best suited for the specific separation relies on the one-at-a-time testing of each individual compound, a tedious process at best. Obviously, the use of a mixed library of chiral additives combined with an efficient deconvolution strategy has the potential to accelerate this selection. 

Lin et al. [95] used capillary electrophoresis with dual cyclodextrin systems for the enantiomer separation of miconazole. A cyclodextrin-modified micellar capillary electrophoretic method was developed using mixture of /i-cyclodextrins and mono-3-0-phenylcarbamoyl-/j-cyclodextrin as chiral additives for the chiral separation of miconazole with the dual cyclodextrins systems. The enantiomers were resolved using a running buffer of 50 mmol/L borate pH 9.5 containing 15 mmol/L jS-cyclodextrin and 15 mmol/L mono-3-<9-phcnylcarbamoyl-/j-cyclodextrin containing 50 mmol/L sodium dodecyl sulfate and 1 mol/L urea. A study of the respective influence of the /i-cyclodcxtrin and the mono-3-(9-phenylcarbamoyl-/i-cyclodextrin concentration was performed to determine the optical conditions with respect to the resolution. Good repeatability of the method was obtained. 

Phinney et al. [Ill] investigated the application of citrus pectins, as chiral selectors, to enantiomer separations in capillary electrophoresis. Successful enantioreso-lution of primaquine and other antimalarials, was achieved by utilizing potassium polypectate as the chiral selector. Changes in pH, chiral additive concentration, and capillary type were studied in relation to chiral resolution. The effect of degree of esterification of pectin materials on chiral recognition was evaluated. 

Ward, T.J., Dann III, C., and Blaylock, A., Enantiomeric resolution using the macrocyclic antibiotics rifamycin B and rifamycin SV as chiral selectors for capillary electrophoresis, J. Chromatogr. A, 715, 337, 1995. 

Schmitt, U., Ertan, M., and Holzgrabe, U. (2004). Chiral capillary electrophoresis facts and fiction on the reproducibility of resolution with randomly substituted cyclodextrins. Electrophoresis 25, 2801-2807. 

Rudaz, S., Calleri, E., Geiser, L., Gherkaoui, S., Prat, J., and Veuthey, J. L. (2003). Infinite enantiomeric resolution of basic compounds using highly sulfated cyclodextrin as chiral selector in capillary electrophoresis. Electrophoresis 24, 2633—2641. 

Capillary electrophoresis offers a set of important advantages that make it a premier technique for the investigation of enantioselective effects in the affinity interactions between chiral drugs and cyclodextrins. The most important advantage of CE is the inherently high separation efficiency offered by this technique. As already known, the most important contributors to peak resolution (R) are a separation selectivity (a) and an efficiency (N). A relationship between these parameters in CE is described by the following equation (2) ... 

D Eberle, RP Hummel, R Kuhn. Chiral resolution of pantoprazole sodium and related sulfoxides by complex formation with bovine serum albumin in capillary electrophoresis. J Chromatogr A 759 185—192, 1997. 

DK Lloyd, A Ahmed, F Pastore. A quantitative relationship between capacity factor and selector concentration in capillary electrophoresis and high-performance liquid chromatography evidence from the enantioselective resolution of benzoin using human serum albumin as chiral selector. Electrophoresis 18 958-964, 1997. 

Among the electrophoretic methods of chiral resolution, various forms of capillary electrophoresis such as capillary zone electrophoresis (CZE), capillary isotachophoresis (CIF), capillary gel electrophoresis (CGE), capillary isoelectric focusing (CIEF), affinity capillary electrophoresis (ACE), and separation on microchips have been used. However, in contrast to others, the CZE model has been used frequently for this purpose [44]. On the other hand, drawbacks associated with the electrophoretic technique due to lack of development of modem chiral phases have limited the application of these methods. Moreover, the electrophoretic techniques cannot be used at the preparative scale, which represents an urgent need of chiral separation science. 

Bonato and Paias [136] developed two sensitive and simple assay procedures based on HPLC and capillary electrophoresis for the enantio-selective analysis of omeprazole in pharmaceutical formulations. Racemic omeprazole and (S)-omeprazole were extracted from commercially available tablets using methanol-sodium hydroxide 2.5 mol/1 (90 10). Chiral HPLC separation of omeprazole was obtained on a ChiralPak AD column using hexane-ethanol (40 60) as the mobile phase and detection at 302 nm. The resolution of omeprazole enantiomers by capillary electrophoresis was carried out using 3% sulfated /1-cyclodextrin in 20 mmol/1 phosphate buffer, pH 4 and detection at 202 nm. 

Contrary to conventional HPLC, almost 98% of chiral resolution in CE is carried out using the chiral selector as a mobile phase additive. Again all the common chiral selectors used in NLC can also be used in NCE. But, unfortunately, few chiral molecules have been tested in NCE for enantiomeric resolution of some racemates. To the best of our knowledge only cyclodextrins and protein-based chiral mobile phase additives have been used for this purpose. Manz and coworkers discussed chiral separations by NCE in their reviews in 2004 [21] and 2006 [22], Later on, Pumera [16] reviewed the use of microfluidic devices for enantiomeric resolutions in capillary electrophoresis. Not much work has been carried out on chiral resolution in NCE but the papers that are available are discussed here. 

The resolution of 3,4-dihydro-2/f-l-benzothiopyran-4-one 1-oxide has been achieved by capillary electrophoresis using heptakis-6-sulfato-/3-cyclodextrin or, better, its 2,3-diacetyl derivative as the chiral selector the i -sulfoxide migrated before the A-enantiomer <2001JSS766>. It has also been noted that (R)-thiochroman 1-oxide is eluted before the (d )-enantiomer from a Chiralcel cellulose tribenzoate column <2001TA1551> and a Daicel Chiralpak AD-H column using hexane/APrOH 90 10 has been used to resolve the (3.y,l. R) 249 and (3/ ,l A) 250 enantiomers of the substituted tetrahydrothiopyran-4-one <20050BC84>. 

Lurie IS, Klein RX, Lebelle M, Brenneisen R, Weinberger R. Chiral resolution of drugs of forensic interest by capillary electrophoresis using cyclodextrins. Anal Chem 1994 66 4019. 

Capillary electrophoresis (CE) provides a valid alternative to HPLC methods for chiral separations. The direct resolution of racemates requires only an enantiomerically pure additive (chiral selector) to be dissolved in the running buffer. The experimental conditions affecting the separations and an overview of practical applications have been compiled <1999ELP2605>. 

Ali, L Gupta, V.K. Aboul-Enein, H. Y, Chiral resolution of racemic environmental pollutants by capillary electrophoresis Crit. Rev. Anal. Chem. 2008, 38, 132-146. 

Kodama, S. et al. Direct chiral resolution of tartaric acid by ion-pair capillary electrophoresis using an aqueous background electrolyte with (lR,2R)-(-)-l,2-diaminocyclo-hexane as a chiral covmterion. Electrophoresis 2003, 24, 2711-2715. 

Analysis of the chiral pollutants at trace levels is a very important and demanding field. In recent years, capillary electrophoresis has been gaining importance in the direction of chiral analysis of various racemates. A search of the literature cited herein indicates a few reports on the chiral resolution of environmental pollutants by CE. It has not achieved a respectable place in the routine chiral analysis of these pollutants due to its poor reproducibility and to the limitations of detection. Therefore many scientists have suggested various modifications to make CE a method of choice. To achieve good reproducibility. 

Fig. 2 Influence of cyclodextrin type on the chiral resolution of the basic drug tocainide and related substances using 40 mM sodium phosphate (pH 3.0) containing (a) 20 mM -CD and (b) 50 mM methyl- -CD and 50 mM heptakis (2,6 di-O-methyl-jS-CD. [Adopted with kind permission from D. Beider and G. Schomburg, Chiral separations of basic and acidic compounds in modified capillaries using cyclodextrin-modified capillary zone electrophoresis, J. Chromatogr. A 666 351 (1994).]...

For enantioseparation on CSPs in CEC, nonstereospecific interactions, expressed as 4>K, contribute only to the denominator as shown in Eq. (1), indicating that any nonstereospecific interaction with the stationary phase is detrimental to the chiral separation. This conclusion is identical to that obtained from most theoretical models in HPLC. However, for separation with a chiral mobile phase, (pK appears in both the numerator and denominator [Eq. (2)]. A suitable (f)K is advantageous to the improvement of enantioselectivity in this separation mode. It is interesting to compare the enantioselectivity in conventional capillary electrophoresis with that in CEC. For the chiral separation of salsolinols using /3-CyD as a chiral selector in conventional capillary electrophoresis, a plate number of 178,464 is required for a resolution of 1.5. With CEC (i.e., 4>K = 10), the required plate number is only 5976 for the same resolution [10]. For PD-CEC, the column plate number is sacrificed due to the introduction of hydrodynamic flow, but the increased selectivity markedly reduces the requirement for the column efficiency. 

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