Enantioselective separations

Among the existing separation techniques, some - due to their intrinsic characteristics - are more adapted than others to processing large amounts of material. Such processes, which already exist at industrial level, can be considered in order to perform an enantioselective separation. This is the case for techniques such as distillation and foam flotation, both of which constitute well-known techniques that can be adapted to the separation of enantiomers. The involvement of a chiral selector can be the clue which changes a nonstereoselective process into an enantioselective one. Clearly, this selector must be adapted to the characteristics and limitations of the process itself. 

All enantioselective separation techniques are based on submitting the enantiomeric mixture to be resolved to a chiral environment. This environment is usually created by the presence of a chiral selector able to interact with both enantiomers of the mixture, albeit with different affinities. These differences in the enantiomer-selector association will finally result in the separation that is sought. 

Optically active polymers are potentially very useful in areas such as asymmetric catalysis, nonlinear optics, polarized photo and electroluminescence, and enantioselective separation and sensing.26 Transition metal coupling polymerization has also been applied to the synthesis of these polymers.27 For example, from the Ni(II)-catalyzed polymerization, a regioregular head-to-tail polymer 32 was obtained (Scheme 9.17).28 This polymer is optically active because of the optically active chiral side chains. 

Ye J, Wu J, Liu W (2009) Enantioselective separation and analysis of chiral pesticides by high-performance liquid chromatography. Trends Anal Chem 28 1148-1163... 

Obviously, the monolithic material may serve its purpose only if provided with a suitable surface chemistry, which depends on the desired application. For example, hydrophobic moieties are required for reversed phase chromatography, ionizable groups must be present for separation in the ion-exchange mode, and chiral functionalities are the prerequisite for enantioselective separations. Several methods can be used to prepare monolithic columns with a wide variety of surface chemistries. 

Moreover, in-situ copolymerization approaches of polymerizable chiral cin-chonan carbamate selectors have also been shown to be viable straightforward routes to enantioselective separation media. In one approach, polymethacrylate-type monoliths have been fabricated by copolymerization of functional monomers and crosslinker in presence of porogenic solvents [80-85]. They have been utilized mainly for CEC (and will be described in detail later) while they turned out to be less suitable for HPLC application because of a low crosslinking degree. 

This review provides an overview of the literature published to date on macrocyclic antibiotics exploited for enantioselective separations in high-performance liquid chromatography (HPLC). It was not intended as a comprehensive issue on the applications of such antibiotics in sub- and supercritical fluid chromatography (SFC), thin layer chromatography (TLC), capillary electrophoresis (CE), and capillary electrochromatography (CEC). A number of structural properties of the most important macrocyclic antibiotics applied in HPLC enantioseparations are listed in Table 2.1. 

As a general rule, in the case of CSPs featuring hydrophobic pockets, a decrease of mobile phase flow-rate results in an increase of chromatographic resolution (Rs), as a consequence of better stationary phase mass transfer [78]. This change has significant impact mostly in RP mode [17]. In the NP enantioselective separations of two test solutes (4-hexyl-5-cyano-6-methoxy-3,4-dihydro-2-pyridone and... 

Very recently, ethoxynonafluorobutane (ENFB) was evaluated as a safe and environmentally friendly NP solvent for the enantioselective separation of 15 compounds on two commercially available glycopeptides CSPs (vancomycin and teicoplanin), with APCI-MS detection [126]. 

A special mention in the field of enantioselective HPLC separations must be made of chiro-optical detection systems, such as circular dichroism (CD) and optical rotation (OR), which can be also used to circumvent the low UV detectability of chromophore-lacking samples [40, 61]. While sensitivity of chiro-optical detection is not always sufficient to perform enantiomeric trace analysis, the stereochemical information contained in the bisignate spectropolarimetric response is useful in establishing elution order for those compounds not available as single enantiomers of known configuration. An example of application of different online detection systems (UV and CD at 254 nm) in the enantioselective separation of a racemic sulfoxide on a commercially available TAG CSP is reported in Figure 2.12, under NP conditions. 

Aboul-Enein, H.Y. and Serignese, V., Optimized enantioselective separation of clenbuterol on macrocyclic antibiotic teicoplanin chiral stationary phase, J. Liq. Chrom. Rel. Technol, 22, 2177, 1999. 

Dolezalova, M. and Tkaczykova, M., HPLC enantioselective separation of aromatic amino and hydrazino acids on a teicoplanin stationary phase and the enantiomeric purity determination of L-isomers used as drugs. Chirality, 11, 394, 1999. 

Bosakova, Z., Cufinovd, E., and Tes ovd, E., Comparison of vancomycin-based stationary phases with different chiral selector coverage for enantioselective separation of selected drugs in high-performance liquid chromatography, J. Chromatogr. A, 1088, 94, 2005. 

Tesalova, E., Bosdkovi, Z., and Pacakova, V., Comparison of enantioselective separation of A -tert-butyloxycarbonyl amino acids and their non-blocked analogues on teicoplanin-based chiral stationary phase, J. Chromatogr. A, 838, 121, 1999. 

Enantioselective separation by supercritical fluid chromatography (SFC) has been a field of great progress since the first demonstration of a chiral separation by SFC in the 1980s. The unique properties of supercritical fluids make packed column SFC the most favorable choice for fast enantiomeric separation among all of the separation techniques. In this chapter, the effect of chiral stationary phases, modifiers, and additives on enantioseparation are discussed in terms of speed and resolution in SFC. Fundamental considerations and thermodynamic aspects are also presented. 

Cinchona Alkaloid-Derived Enantioselective Separation Materials.. 

When using PFT with a neutral selector, it is quite difficult to avoid any entrance of the chiral selector into the ionization source, particularly at a high pH, where EOF is important. The use of BGE at low pH and/or coated capillary to minimize EOF is therefore mandatory. However, the coaxial sheath gas, which generally assists the ionization process, leads to an aspirating phenomenon of the chiral selector in the MS direction. Javerfalk et al. were the first to apply PFT with a neutral methyl-/i-CD for the separation of racemic bupivacaine and ropivacaine with a polyacrylamide-coated capillary and an acidic pH buffer (pH 3). Cherkaoui et al. employed another neutral CD (HP-/1-CD) with a PVA-coated capillary for the analysis of amphetamines and their derivatives. To prevent a detrimental aspiration effect, analyses were carried out without nebulization pressure. Numerous other studies presented excellent results such as the enantioselective separation of adrenoreceptor antagonist drugs using tandem mass spectrometry (MS/MS) the separation of clenbuterol enantiomers after solid-phase extraction (SPE) of plasma samples or the use of CD dual system for the simultaneous chiral determination of amphetamine, methamphetamine, dimethamphetamine, and p-hydroxymethamphetamine in urine. 

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