Chiral separation of dansyl amino acids by capillary electrophoresis: Comparison of formamide andN-Methylformamide as background electrolytes

Summary Chiral separation of 12 dansyl amino acids has been achieved by capillary electrophoresis using -cyclodextrin dissolved inN-methylformamide or formamide. The viscosity and the dielectric constants of these two solvents are very different, giving them disimilar electrophoretic properties. Intense electroosmotic flow, high electrophoretic mobilities and high efficiency were achieved inN-methylformamide, but high chiral selector concentration was needed for sufficient resolution. In contrast, the separation of dansyl amino acids in formamide was characterized by longer analysis time and lower efficiency, but the resolution and selectivity of the separation were better and baseline separation could be achieved at lower -cyclodextrin concentrations.     

Novel chiral solvating agents derived from natural amino acid: enantiodiscrimination for chiral α-arylalkylamines

Two benzo[de]isoquinoline 1,3-dione amino acids 1 and 2 were readily prepared, and their enantiodiscriminating ability were investigated by ¹H NMR spectroscopy. It was found that 1 exhibited an excellent chiral recognition ability toward chiral α-phenylethylamine and some of its derivatives, leading to clear baseline separation of the multiplet of the probe groups in two enantiomers. The stoichiometric ratio and association constants of some host-guest complexes were determined. The interactions between the hosts and guest 3 were further studied by intermolecular NOE experiment and ESI-MS.     

Enantioseparation of α-phenylglycine by HPLC on an ODS column coated with chiral crown ether

Summary Previous work has demonstrated the possibility of crcating chiral stationary phases by coating a suitable support material via the mobile phase. The result is a wide variety of chiral phase systems which can be used in liquid chromatogrphy to separate enantiomers by a liquid-solid adsorption mechanism. The present paper demonstrates the potential of this technique for chiral crown ethers incorporating an -D-mannopyranoside unit using the separation of phenyglycine enantiomers as an example.     

Chiral separation on achiral stationary phases with different functionalities using β-cyclodextrin in the mobile phase and application to bioanalysis and coupled columns

Summary -cyclodextrin was used in the mobile phase as chiral selector for separating the enantiomers of terbutaline, chlorthalidone and oxazepam. The effect on chiral resolution using e.g. hydrophobic, polar or cation exchanging stationary phases was investigated. Both the chiral separation factor and retention level were affected by the concentration of methanol and -cyclodextrin. The stationary phase had no effect on the chiral separation only on the level of retention. By tuning the concentration of -cyclodextrin and methanol in the mobile phase chiral separation could be obtained on most stationary phases. By changing the stationary phase while adjusting the mobile phase composition to maintain the chiral selectivity, improvements of the selectivity towards e.g. endogenous compounds can be obtained when separating enantiomers in complex matrixes as biological fluids. Further improvement on selectivity can be obtained if coupled columns are used. This is examplified for separation of chlorthalidone and terbutaline enantiomers in biological fluids by coupling an achiral column to another achiral column and using a mobile phase containing -cyclodextrin on the last column.     

Chiral Resolution of Enantiomers of Homocamptothecin Derivatives, Antitumor Topoisomerase I Inhibitors, Using High Performance Liquid Chromatography on Polysaccharide-Based Chiral Stationary Phases

Analytical HPLC methods using derivatized cellulose and amylose chiral stationary phases were developed for the separation of the enantiomers of homocamptothecin (hCPT) derivatives which constitute a promising series of potent anticancer agents targeting DNA topoisomerase I. The resolutions were performed using a normal phase methodology with two silica-based celluloses tris-3,5-dimethylphenylcarbamate (Chiralcel OD-H) and tris-methylbenzoate (Chiralcel OJ) or two amyloses tris-3,5-dimethylphenylcarbamate (Chiralpak AD) and tris-(S)-1-phenylethylcarbamate (Chiralpak AS). The mobile phase and the chiral stationary phase were varied to achieve the best resolution. Different types and concentration of aliphatic alcohols in the mobile phase were also tested along with the temperature dependence. An optimal baseline separation (Rs > 1.5) was readily obtained in most cases. The different columns gave complementary results in term of resolution. The limits of detection and quantification were between 0.08–0.40 M and 0.24–1.80 M, respectively and the enantiomeric purity was superior to 99.9%.     

The influence of end-capping on the enantioselectivity of a chiral phase

Summary The effect of end-capping chiral stationary phases (CSP's) derived fromN-(2-naphthyl)alanine undecyl ester has been examined using either trimethylchlorosilane (TMCS), hexamethyldisilazane (HMDS), or bis(trimethylsilyl) trifluoroacetamide (BSTFA) as end-capping reagents. The separation factor () and capacity factor (k) of the enantiomers ofN-(3,5-dinitrobenzoyl)leucine octadecyl amide andN-(3,5-dinitrobenzoyl)alanine butyl ester were evaluated on three columns all packed with material from the same batch of stationary phase. These columns were essentially identical before, but not after end-capping with the above reagents. TMCS and HMDS were found to be superior to BSTFA, which appears to cause a significant loss of bonded phase from the silica surface. It seems that residual silanols affect the retention either by interacting with the analyte or by interacting with strands of stationary phase. End-capping usually increases enantioselectivity, sometimes by decreasing k for the first enantiomer and increasing k for the second enantiomer. The enhancement in enantioselectivity is greatest in relatively nonpolar mobile phases and occurs to a greater extent for phases having incomplete surface coverages.     

Optimized Separation of cis-trans Isomers and Enantiomers of Sertraline Using Cyclodextrin-Modified Micellar Electrokinetic Chromatography

Micellar electrokinetic chromatography has been investigated for the separation of cis-trans isomers and enantiomers of sertraline. The effects of various separating factors were studied. Optimum separation was achieved using a buffer (pH 11.5) of 35 mM sodium borate containing 30 mM sodium deoxycholate and 20 mM hydroxypropyl –-cyclodextrin; the optimum voltage and temperature were 25 kV and 20 °C, respectively. A detection wavelength of 210 nm was used. The analytical performance of the method was discussed in terms of linearity response, precision, detection limits, quantification limits and recoveries. The method developed was successfully applied to the determination of sertraline in bulk drug, tablets and capsules.Revised: 1 April and 18 May 2004     

Separation of Enantiomers of Non-Protein Amino Acids by High-Performance Liquid Chromatography on a Chiral Ligand-Exchange Column

The HPLC separation of enantiomers of underivatized non-protein amino acids was investigated by using a column packed with octadecylsilanized silica coated with N,S-dioctyl-D-penicillamine as a chiral ligand-exchange phase (Sumichiral OA-5000). Good enantiomeric separations were achieved with a variety of -amino acids carrying aliphatic or aromatic side chains, cyclic imino acids, and -amino acids, together with -methyl--amino acids, by optimizing the amount (0–20%, v/v) of 2-propanol as the organic component and the concentration (1–5 mM) of Cu²⁺ as the complexing metal ion in the aqueous-organic eluent.     

The n→π* optical activity of chiral carbonyl compounds predicted to be optically inactive by the octant rule

The one-electron model of n* optical activity in chiral carbonyl compounds is carried to second-order in perturbation theory and the results are applied to systems which are predicted by the octant rule to yield zero n* optical activity. It is shown that second-order contributions to the n* rotatory strength lead to sector rules which are qualitatively different from those obtained from first-order contributions. Inclusion of three-way interactions involving the carbonyl chromophore and two different extrachromophoric perturbing groups (or atoms) lead to second-order contributions to the rotatory strength which can account for the n* optical activity in chiral carbonyl systems predicted to be optically inactive by the conventional quadrant and octant rules. These results are shown to be in agreement with the conclusions derived by Ruch and Schönhofer from a purely algebraic theory of molecular chirality.     

Simultaneous chiral separation of leucovorin and its major metabolite 5-methyl-tetrahydrofolate by capillary electrophoresis using cyclodextrins as chiral selectors: Estimation of the formation constant and mobility of the solute-cyclodextrin complexes

Summary Capillary electrophoresis with an electrolyte containing cyclodextrin was investigated for the simultaneous separation of the diastereoisomers of 6R,S-leucovorin and its active metabolite 6R,S-5-methyl-tetrahydrofolate. , and -cyclodextrin separated the diastereoisomers of 5-methyl-tetrahydrofolate, while only -cyclodextrin was found to be effective for the chiral separation of leucovorin. The effect of -cyclodextrin concentration was investigated, and subsequently a curve-fitting analysis for the quantitative estimation of the binding constants was attempted. The binding constants were found to be very small, in the range 2–4 M^–1. Although the interaction between -cyclodextrin and the tetrahydrofolates is weak, the high efficiency of capillary electrophoresis and the use of high concentrations of -cyclodextrin allow baseline chiral separation of the diastereoisomers of leucovorin and 5-methyl-tetrahydrofolate. Changes in temperature exert differing effects on the separations of leucovorin and 5-methyl-tetrahydrofolate; higher temperatures improved the separation of leucovorin diastereoisomers but reduced the resolution of 5-methyl-tetrahydrofolate diastereoisomers. The effects of urea and buffer salt concentrations and of buffer pH were also investigated. Capillary electrophoresis with -cyclodextrin was used to analyse plasma samples spiked with clinically-relevant levels of leucovorin and 5-methyl-tetrahydrofolate. Resolution of these compounds in ultrafiltered plasma was demonstrated, but detection sensitivity was not adequate for the routine use of this method for the determination of leucovorin and 5-methyl-tetrahydrofolate in plasma. In addition, a simple technique to reverse the elution order of ionic stereoisomers was demonstrated. By adding a cationic surfactant into the buffer and reversing the separation potential, the elution order of the diastereoisomers of leucovorin and 5-methyl-tetrahydrofolate was reversed.     

Use of inorganic, complex-forming ions for selectivity enhancement in capillary electrophoretic separation of organic compounds

Capillary electrophoresis (CE) is a powerful separation method based on the migration of charged species under the influence of electric field. The main merits of CE are high separation efficiency, short analysis time and small consumption of solvents and samples. However, the main drawbacks of CE are generally lower sensitivity compared to classical column-chromatographic methods.Selectivity and/or sensitivity of CE separation can be improved by forming complexes between analytes and a complex-forming reagent present as an additive in the background electrolyte (BGE). We focus this review primarily on the application of inorganic complex-forming reagents added to the BGE to separate organic ligands. We briefly mention common CE separations of inorganic analytes (mainly metal ions) using BGEs with organic ligands (e.g., hydroxycarboxylic or aminopolycarboxylic acids) as selectors.The review involves brief theoretical consideration of the significance of the effect of complex formation on separation selectivity and/or sensitivity in CE, but the major topic is critical evaluation of different inorganic complex-forming reagents used recently in the CE analysis of organic compounds, including:

(i)

borate, tungstate and molybdate in separating organic compounds possessing vicinal -OH groups;

(ii)

ligand-exchange CE and capillary electrochromatography in chiral analysis; and,

(iii)

the role of metal ions as central ions employed for selectivity enhancement of CE separation of various classes of organic compounds, including biopolymers.

     

Asymmetric synthesis of α-fluoro-α-sulfenyl-β-ketoesters using DBFOX-Ph/Ni(II) complex

Enantioselective α-sulfenylation of α-fluoro-β-ketoesters 4 with phenylsulfenyl chloride catalyzed by DBFOX-Ph/Ni(II) complex afforded the corresponding α-fluoro-α-sulfenyl-β-ketoesters 2 in moderate to good yields with excellent enantiomeric excesses up to 93% ee. α-Fluoro-α-sulfenyl-β-ketoesters can be effectively converted to tri-fluorinated α-sulfenylcarboxylates by the use of DAST, which should be useful intermediates for the synthesis of non-racemized fluorinated isosteres of pharmaceutically attractive SM₃₂. The enantioselective α-phenylsulfenylation as well as α-pentafluoro-phenylsulfenylation of non-fluorinated β-ketoesters 5 were also carried out under the same catalyst conditions affording up to 95% ee of the products 6-8.     

Chiral Separation of Racemates of Drugs and Amino Acid Derivatives by High-Performance Liquid Chromatography on a Norvancomycin-Bonded Chiral Stationary Phase

A novel norvancomycin-bonded chiral stationary phase (NVC-CSP) has been synthesized by use of the chiral selector norvancomycin, which differs from vancomycin because of the presence of leucine rather N-methylleucine. The enantiomers of some neutral and basic chiral drugs, for example warfarin, benzoin, bendroflumethiazide, and praziquantel, were directly separated by high-performance liquid chromatography in the reversed-phase mode. The effect of conditions such as organic modifier concentration, column temperature, pH, and mobile phase flow rate on retention and enantioselectivity were investigated. It was shown that hydrophobic, steric, and ionic interactions were present between the analyte and the macrocycle in this chromatographic system. Vant Hoff plots afforded the thermodynamic data _R,SH° and _R,SS°; the negative values obtained indicated the process of enantiomer separation was enthalpy-controlled. In an attempt to improve the resolution of some very polar acidic compounds (dansyl-amino acids) norvancomycin was used as stationary phase chiral selector and chiral mobile-phase additive simultaneously, better results were obtained as the result of a synergistic effect. It was also shown experimentally that the newly synthesized NVC-CSP behaved somewhat differently from the earlier reported vancomycin-bonded CSP, probably because of the different structures of norvancomycin and vancomycin.     

Chiral separations by host-guest complexation with cyclodextrin and crown ether in capillary zone electrophoresis

Summary Capillary zone electrophoresis using cyclodextrins and a chiral crown ether as buffer constituents was studied for the enantiomeric separation of drugs and amino acids. Based on results obtained from separation of racemic -amino acids both chiral selectors are compared with respect to resolution, efficiency and retention time. For (±)-Quinagolide effects of buffer composition and temperature are examined using -cyclodextrin as chiral agent. Optimum conditions were pH 2.5 at 30 mmol L^–1 -cyclodextrin. A linear dependence of retention on -cyclodextrin concentration allowed calculation of formation constants of the host-guest complexes. Buffer concentration and temperature also influence resolution. The application of a chiral crown ether to the separation of optical isomers in capillary zone electrophoresis is described for the first time. Chiral recognition of solutes depends on the formation of protonated alkyl amines and separation is attributed to the formation of diastereomeric host-guest complexes with different interactions for each enantiomer. The effects of crown ether concentration on resolution are presented.     

On the influence of chiral auxiliaries in the stereoselective cross-coupling reactions of titanium enolates and acetals

Titanium enolates from chiral N-propanoyl-1,3-thiazolidine-2-thiones containing bulky substituents at C4 turned out to be excellent platforms to get highly stereocontrolled cross-coupling reactions with acetals. Related oxazolidinethiones also afforded good results, but the corresponding oxazolidinones resulted completely unselective for such reactions, which proves that an exocyclic CS bond is essential to attain a synthetically useful stereocontrol.     

Separation of Enantiomers of N-Protected Non-Protein Amino Acid Esters by Chiral High-Performance Liquid Chromatography

The high-performance liquid chromatographic separation of enantiomers of N-protected non-protein amino acid esters was investigated by using a cellulose tris(3,5-dimethylphenylcarbamate) chiral stationary phase column (Daicel Chiracel OD). The effect of the N-protecting groups and the ester groups on chiral discrimination was examined. The benzyloxycarbonyl (Z), 4-methoxybenzyloxycarbonyl, and 9-fluorenylmethoxycarbonyl derivatives gave good enantiomeric separations, while the formyl and t-butoxycarbonyl groups marred them. Almost all the alkyl esters examined and the benzyl ester gave enantiomeric separations better than or of the same order as the methyl ester. The N-Z-protected methyl esters of a number of non-protein -amino acids were well resolved using hexane–2-propanol as a mobile phase. The resolution of -amino acid derivatives was inferior to that of the isomeric -amino acid derivatives.     

Diastereoselective and enantioselective chromatography of the pyrethroid insecticides allethrin and cypermethrin

Summary Liquid and gas chromatographic separations of the pyrethroid insecticides allethrin and cypermethrin have been investigated with various achiral and chiral stationary phases. Diastercomeric and enantiomeric selectivity was observed for cypermethrin on a Pirkle-type chiral LC stationary phase, but very strong interactions and therefore long retention times prevented the separation of allethrin on this phase. Trans-allethrin isomers were separated on a chiral -cyclodextrin RP-HPLC column while cypermethrin showed some difficulties on this phase due to isomerization. Diastereomeric but no enantiomeric selectivity by GC was achieved for cypermethrin with an apolar DB 5 capillary. GC separation of the diastereomers was used to study the selective photodegradation of cypermethrin isomers after forestry applications. Chiral -cyclodextrin-based GC phases showed some enantioselectivity for cis- and trans-allethrin isomers. A separation of the eight isomers into six partially resolved peaks was achieved by GC with a coupled column consisting of chiral permethylated -cyclodextrin and DB 1701 as stationary phases. This combination was used to characterize allethrin formulations intended for indoor use and to investigate allethrin products formed by ozonolysis of thin films of the insecticide.     

High-performance liquid chromatographic resolution of enantiomers on chiral amine-bonded silica gel

Summary A chiral stationary phase with an immobilized, optically active diamine was prepared for the separation of enantiomers. The synthesis of the phase was carried out by bonding (–)trans-1,2-cyclohexanediamine to microparticulate silica gel through the coupling agent 3-glycidoxypropylsilane. The resolution of the racemic compounds catechin, 2,2-dihydroxy-1,1binaphthyl and trans-1,2-cyclohexandiol, is reported.     

Enantiomeric separation of amphetamine,methamphetamine and ring substituted amphetamines by means of a β-cyclodextrin-chiral stationary phase

Summary The enantioseparation of amphetamine, methamphetamine and various ring-substituted amphetamines by use of a chiral stationary phase carrying immobilized native -cyclodextrin (-CyD) selectors is reported. The system is evaluated for resolving the specified compounds directly without any derivatization and after derivatization with phenyl isothiocyanate (PITC), naphthyl isothiocyanate (NITC) and 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). This direct enantioseparation is compared with the features of indirect separation of diasteromeric derivatives after reaction with the optically pure Marfey's reagent employing a simple non-chiral alkyl-silica (RP-8) column. A selection of those methods best suited for each single amphetamine is given.Seventeen different samples of amphetamine, confiscated by the Swedisch police, were analyzed with respect to their enantiomeric composition. Within this set of samples synthesized by the same method no significant deviation from a racemic ratio could be observed.