Liquid chromatography chromatographic resolution

Using amylose tris-3,5-dimethylphenylcarbamate as the chiral selector in enantioselective high-performance liquid chromatography, micropreparative resolution of the DHA racemate was achieved and the chromatographic behaviour in enantio-GC could be defined by coinjecting these references of definite chirality (Fig. 17.4) [13]. 

Resolution can often be improved in liquid chromatographic separations simply by changing the retention of the components, which corresponds to changing the capacity factor k of the components to be separated. Rapid increases in the third term in Eq. (5), 2/(1 + 2). occur as the capacity factor increases, up to 2 = 5. The term increases only slowly beyond this value [2]. Therefore, in most separations by liquid chromatography, optimal resolution of mixtures occurs when each component has capacity factor values between 2 and about 10. Increasing k values to higher values results in longer analysis times with minimal improvements in resolution [2]. 

HPLC separations are one of the most important fields in the preparative resolution of enantiomers. The instrumentation improvements and the increasing choice of commercially available chiral stationary phases (CSPs) are some of the main reasons for the present significance of chromatographic resolutions at large-scale by HPLC. Proof of this interest can be seen in several reviews, and many chapters have in the past few years dealt with preparative applications of HPLC in the resolution of chiral compounds [19-23]. However, liquid chromatography has the attribute of being a batch technique and therefore is not totally convenient for production-scale, where continuous techniques are preferred by far. 

Thick-layer silica gel chromatography can also be employed [7], although most separations are now accomplished by high-performance liquid chromatography. Resolution of complex mixtures often requires both normal and reverse phase modes [19]. A further dimension is added, when bioactivity is correlated with spectroscopically-monitored chromatographic profiles. 

Total resolution of [269] into its two enantiomers was achieved by liquid-liquid chromatography through complexation to L-valine adsorbed initially on diatomaceous earth (Timko et al., 1978). On the basis of comparative chromatographic studies, the separation factors (a) and the EDC values were correlated (Cram et al., 1975) (Table 59). 

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