Chiroptical detection

Further, peak overlap results in nonlinear detector response vs concentration. Therefore, some other detection method must be used in conjunction with either of these types of detection. Nevertheless, as can be seen from Figure Ilf, chiroptical detection can be advantageous if there is considerable overlap of the two peaks. In this case, chiroptical detection may reveal that the lea ding and tailing edges of the peak are enantiomerically enriched which may not be apparent from the chromatogram obtained with nonchiroptical detection (Fig. He). 

Fig. 11. Simulated chromatograms of chiral separations obtained using nonchiroptical detection (a, c, e) and chiroptical detection (b, d, f) illustrating the...

Xanthan is reported to undergo a chiroptically detected temperature or salt-driven conformational change from an ordered conformation at high salt and low temperature to a disordered conformation either associated with lowering the salt concentration, or with increasing the temperature (2-5). The primary structure of xanthan has been known for about a decade (6,7), but different structures have been suggested both for the ordered and disordered conformation. Some workers (8-13) conclude that the ordered conformation is double-stranded or double-helix, whereas others (14-17) claim that a single stranded description can account for the observed data under... 

A. Mannschreck, Chiroptical detection during liquid chromatography, Chirality, 4 163 (1992). 

To be effective, the light sources used for chiroptical detection systems must have an intensity that is much greater than those ordinarily used in polarimetry. This comes about because the angular rotations observed for the very low analyte concentrations and very short sample pathlengths typical of an analytical liquid chromatograph, are extremely small (mdeg. and less). Conventional light sources have been replaced with laser illumination but these are not without problems, a major one of which is the instability in the emission [23],... 

CD detection for colored derivatives might just as easily be used for structural information about the carbohydrates as well as for their analysis, but so far little attention has been given to either application. Cyclic oligomers of p-D-glucose have important supporting roles to play in analytical applications that are discussed in a later section. The union of chirality in the carbohydrate moiety of a glycoside metabolite with the unsaturation in the base in such compounds as nucleosides and nucleotides, saponins and flavones, etc., is another area that will ultimately be developed for applications of chiroptical detection methods. New and imaginative ideas are needed for the analysis of carbohydrates, and CD should be one of the favored methods. 

Cellulose, starch, and their derivatives are commonly used as chromatographic stationary phases. They are, in principle, potential hosts for inducing CD activity in small molecules and could be used with effect for analysis in homogeneous media with chiroptical detection. An example might be the starch (amylose)-iodide complex [86]. Low aqueous solubility however is an obstacle to their general use in homogeneous solutions. Linear oligomers of maltose are more soluble than starch and could theoretically be used as alternatives to Cy, however they do not really compete in terms of the stability of the association complexes. 

The fine structure of the absorption band associated with the n-7T electronic transitions in A -3-ketosteroids is better resolved using chiroptical detection methods compared to absorbance spectrophotometry. Selectivity in the determination of these steroids, therefore, is increased using the solvent-dependent CD spectra of the compounds being investigated, and enhanced even more if difference CD procedures are developed [24],... 

Miiller, M.D. Buser, H.-R., Identification of the (+)- and (—)-enantiomers of chiral chlordane compounds using chiral high-performance liquid chromatography/chiroptical detection and chiral high-resolution gas chromatography/mass spectrometry Anal. Chem. 1994, 66, 2155-2162. 

D2 and D3 vitamins (ergocalciferol and cholecalci-ferol) has not been equally successfulJ Vitamin D extracted from natural sources has a single conformational stereochemistry that is one of several isomers produced in synthetic preparations. To certify that the natural form is present in a synthetic product, where it can be accurately assayed in the presence of the other isomers, is a formidable analytical task. Whether direct CD detection can satisfactorily solve it is currently unknown. A prior non-selective derivati-zation reaction might be required on all isomers. The A and E vitamins are achiral and not subject to chiroptical detection unless first derivatized by reaction with a chiral host. 

The seminal work on steroid analyses using chiroptical detection was done by Djerrasi by the determination of hecogenin acetate in the presence of tigonenin acetate.Every steroid is chiral and therefore amenable to polarimetric detection after chromatographic separation. Chromophores are fairly uncommon, and analysis by ORD or CD is therefore less suitable. The only unsaturation in the cholesterol molecule, for example, is the isolated A -double bond, which has an absorbance maximum at 205 nm. Unsaturation coupled with chirality provides some selectivity, as ably demonstrated by the work of Potapov for analogs of progesterone Even simpler than that is the direct discrimination between the ketosteroids testosterone and dihydrotestosterone, which have opposite signs in methylene chloride solution (Fig. 6). 

The other extreme is an analytical chromatogram with no sign of separation at all, as far as photometric detection is concerned. Nevertheless, the corresponding chiroptical detection often indicates some degree of enantioselectivity (cf. the polarimetric curve for the first run in Figure 3). In such cases, recycling33 the racemate several times through the same column increased the separation, as in... 

Preparative enrichment of enantiomers should be followed by determination of their chemical and enantiomeric purities, e.g., by enantioselective liquid chromatography.25,26 In addition to the sorbents mentioned above, others may be used which are available in only smaller amounts. In our laboratory, this is true for (+)-poly(trityl methacrylate) on silica,35 which can be used for analytical purposes. Preparative separations on a column 0.5 cm in diameter, however, would require many injections of small amounts of racemate. In the case of a baseline chromatogram like that in Figure 2, the determination of enantiomeric purity by measurement of the two peak areas of the enantiomers is straightforward. An analytical chromatogram showing some separation by photometric detection in spite of peak overlap (Figure 4) can still be used for optical purity determina-tion. The simultaneous use of both photometric and chiroptical detection as... 

Kiesswetter R, Brandi F, Kasmer-Pustet N, and Mann-schreck A (2003) Chiroptical detection during liquid chromatography deconvolution of overlapping peaks of enantiomers and its applications. Chirality 15 40-49. 

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