Cyclodextrins, modified, chiral separation

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. 

Anions and uncharged analytes tend to spend more time in the buffered solution and as a result their movement relates to this. While these are useful generalizations, various factors contribute to the migration order of the analytes. These include the anionic or cationic nature of the surfactant, the influence of electroendosmosis, the properties of the buffer, the contributions of electrostatic versus hydrophobic interactions and the electrophoretic mobility of the native analyte. In addition, organic modifiers, e.g. methanol, acetonitrile and tetrahydrofuran are used to enhance separations and these increase the affinity of the more hydrophobic analytes for the liquid rather than the micellar phase. The effect of chirality of the analyte on its interaction with the micelles is utilized to separate enantiomers that either are already present in a sample or have been chemically produced. Such pre-capillary derivatization has been used to produce chiral amino acids for capillary electrophoresis. An alternative approach to chiral separations is the incorporation of additives such as cyclodextrins in the buffer solution. 

Based on the theory, the separation of enantiomers requires a chiral additive to the CE separation buffer, while diastereomers can also be separated without the chiral selector. The majority of chiral CE separations are based on simple or chemically modified cyclodextrins. However, also other additives such as chiral crown ethers, linear oligo- and polysaccharides, macrocyclic antibiotics, chiral calixarenes, chiral ion-pairing agents, and chiral surfactants can be used. Eew non-chiral separation examples for the separation of diastereomers can be found. 

G Galaverna, R Corradini, A Dossena, R Marchelli. Histamine-modified cationic /3-cyclodextrins as chiral selectors for the enantiomeric separation of hydroxy acids and carboxylic acids by capillary electrophoresis. Electrophoresis 20 2619-2629, 1999. 

CL Cooper, JB Davis, MJ Sepaniak. Mechanisms of enantiomeric resolution in cyclodextrin-modified capillary electrophoretic separations of binaphthyl compounds. Chirality 7 401-408, 1995. 

Capillary gas chromatography (GC) using modified cyclodextrins as chiral stationary phases is the preferred method for the separation of volatile enantiomers. Fused-silica capillary columns coated with several alkyl or aryl a-cyclo-dextrin, -cyclodextrin and y-cyclodextrin derivatives are suitable to separate most of the volatile chiral compounds. Multidimensional GC (MDGC)-mass spectrometry (MS) allows the separation of essential oil components on an achiral normal phase column and through heart-cutting techniques, the separated components are led to a chiral column for enantiomeric separation. The mass detector ensures the correct identification of the separated components [73]. Preparative chiral GC is suitable for the isolation of enantiomers [5, 73]. 

Chiral mobile-phase modifiers 133 Separation of dansylated amino acids using /3-cyclodextrin in mobile phase and reversed-phase column... 

The preparative-scale separation of enantiomers on chiral stationary phases (CSPs) by GC cannot match the overwhelming success achieved in the realm of liquid chromatography (LC) (Francotte, 1994, 1996 and 2001). Modern commercial instrumentation for preparative-scale GC is not readily available. In contrast to LC, separation factors a in enantioselective GC are usually small (a = 1.01 - 1.20). This is beneficial for fast analytical separations but detrimental to preparative-scale separations. Only in rare instances are large chiral separation factors (a > 1.5) observed in enantioselective GC. Only in one instance, a separation factor as high as a = 10 was detected in enantioselective GC for a chiral fluorinated diether and a modified 7-cyclodextrin (Schurig and Schmidt, 2003) (vide supra). 

Barbiturates Permethyl-(3-cyclodextrin-modified silica gel (Nucleosil, 5 pm) Methanol-5 mM phosphate buffer, pH 7.0 (1 4) 400 mm x 100 pm i.d. 235 mm effective length, chiral separation, pressure-supported CEC 139... 

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. 

CE is playing a major role in the separation of chiral compounds, a field that is gaining increasing attention in pharmaceutical sciences as well as in forensic toxicology (Lurie, 1994 Novotny et al., 1994 Ward, 1994). The chirally active selectors used in CE include optically active complexes such as Cu(II)-l-histidine, Cu(II)-aspartame, cyclodextrins, modified CDs, bile salts, crown ethers, and proteins (bovine serum albumin, aracid glycoprotein, etc.). 

The first separations of enantiomers in GC on cyclodextrin modified column were carried out by Sybilska et al. in 1983 [5], They applied a formamide solution of a-cyclodextrin as a stationary phase in the classical packed column. The column allowed an efficient separation of chiral monoterpenes - a- and P-pinenes into enantiomers. This system of using CDs in GC is characterised by obtaining high enantioselectivity factors, so enantioseparation is still possible for receiving not very efficient packed columns. Unfortunately, the columns appeared to be not very stable at higher temperatures. 

Several other stationary phases made from different proportions of typical phases (methyl, phenyl, cyanopropyl), or from special compounds such as polytrifluropropyl-siloxane, or different columns such as PLOT (porous layer open tubular), columns coated with a modified graphitized carbon or with a silicone based polymer with chiral groups incorporated into the polymeric chain, columns coated with derivatized cyclodextrins (for the separation of chiral compounds), etc. are also utilized. 

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).]...

T. Ueda, F. Kitamura, R. Mitchell, T. Metcalf, T. Kuwana, and A. Nakamoto, Chiral separation of naph-thalene-2,3-dicarboxaldehyde-labeled amino-acid enantiomers by cyclodextrin-modified micellar electro-kinetic chromatography with laser-induced fluorescence detection, Chem. 63 2919 (1991). 

As shown in Eq. (2) together with the chiral recognition Kk + Kg), the other necessary requirement for enantioseparations in CE is a mobility difference between the free and the complexed analyte fXf - fjL, 0). Otherwise, it will be impossible to transfer a chiral recognition into a chiral separation. This requirement does not hold when neutral analytes are analyzed with neutral chiral selectors. In such a case, an additional buffer component is required that will assist in generating a difference between the mobilities of an analyte in its free and complexed forms with a chiral selector. This is achieved by an achiral micellar phase in cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) [9]. However, a charged CD or a chiral micellar phase can combine the... 

Furuta R, Doi T. 1994. Chiral separation of diniconazole, uniconazole and structurally related compounds by cyclodextrin-modified micellar electrokinetic chromatography. Electrophoresis 15 1322-1325. 

Marina, M.L., I. Benito, J.C. Dfez-Masa, and M.J. Gonzalez. 1996. Chiral separation of polychlorinated biphenyls by micellar electrokinetic chromatography with y-cyclodextrin as modifier in the separation buffer. Chromatographia 42 269-272. 

Methods development for chiral analyses has been one of the most challenging separation problems for the analytical chemist in the pharmaceutical industry. Racemic drug substances have a variety of chemical structures and several chiral selectors are available for the analyst to choose in order to obtain the enan-tioselectivity needed for chiral resolution. To alleviate this problem, a fast capillary electrophoresis procedure for the enantiomeric separation of acidic and basic compounds using native and modified cyclodextrins has been described (200). The technique is called cyclodextrin array chiral analysis. A generalized optimi-... 

L Liu, MA Nussbaum. Systematic screening approach for chiral separations of basic compounds by capillary electrophoresis with modified cyclodextrins. J Pharm Biomed Anal 19 679-694, 1999. 

Nishi, H. Fukuyama, T. Terabe, T. Chiral separation by cyclodextrin-modified micellar electrokinetic chromatography. J. Chmmatogr. 1991, 553, 503-516. 

Nielsen, M. W. F. Chiral separation of basic drugs using cyclodextrin-modified capillary zone electrophoresis. 

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