Enantiomer discrimination by mass spectrometry: noncovalent interactions of an N-derivatized dipeptide with various cinchona alkaloid derivatives and comparison with enantioselective liquid-phase separations

The enantiomer discrimination properties of cinchona alkaloid derived chiral selectors (CSs) towards a dipeptide analyte are examined by electrospray ionization mass spectrometry. The complexes formed between the CSs and the analyte enantiomers owing to various noncovalent interactions are analyzed and the magnitudes of enantiomer discrimination are determined from the complexes mass spectrometric intensities. The influence of different structural features of the CSs on enantioselectivity is discussed. The enantiomer discrimination results obtained by mass spectrometry are compared with those from related liquid chromatography enantiomer separations. A certain coherence between the chromatographic and mass spectrometric enantioselectivities could be established and the enantiomer discrimination patterns, i.e., the relative binding strengths, were identical for the two techniques. Thus, the use of mass spectrometry as a screening tool in the development of new CSs for chromatographic applications seems feasible.     

Stereoelective polymerization of tert-butyl thiirane

The polymerization in bulk of racemic tert-butyl thiirane with a chiral initiator resulting from the reaction between diethylzinc and (? )3,3-dimethyl-1,2-butanediol produces an optically active polymer by preferential consumption of R enantiomer. The unreacted monomer is enriched in S enantiomer. The relative rate r of consumption of R enantiomer versus S enantiomer is as high as 2.8. Obtained polymer could be separated into two crystalline fractions: an optically active fraction, formed from regular sequences of R type enantiomeric units, and an optically inactive fraction which corresponds to a racemate. Experimental results are consistent with a stereospecific mechanism of addition, the two enantiomers being chosen by two different type of sites. The stereoelective process is due to an unequal number of these two types of sites.     

Method Development of Enantiomer Separations by Affinity Capillary Electrophoresis,Cyclodextrin Electrokinetic Chromatography and Capillary Electrophoresis-Mass Spectrometry

 Capillary electrophoresis (CE) has become a powerful tool for enantiomer separations during the last decade. Since 1993, the author has investigated enantiomer separations by affinity capillary electrophoresis (affinity CE) with some proteins and by cyclodextrin electrokinetic chromatography (CDEKC) with some charged cyclodextrins (CDs). Many successful enantiomer separations are demonstrated from our study in this review article. In the enantiomer separations by affinity CE, the deterioration of detection     

Enantiomer analysis of chiral carboxylic acids by AIE molecules bearing optically pure aminol groups

Pure enantiomers of carboxylic acids are a class of important biomolecules, chiral drugs, chiral reagents, etc. Analysis of the enantiomers usually needs expensive instrument or complex chiral receptors. However, to develop simple and reliable methods for the enantiomer analysis of acids is difficult. In this paper, chiral recognition of 2,3-dibenzoyltartaric acid and mandelic acid was first carried out by aggregation-induced emission molecules bearing optically pure aminol group, which was easily synthesized. The chiral recognition is not only seen by naked eyes but also measured by fluorophotometer. The difference of fluorescence intensity between the two enantiomers of the acids aroused by the aggregation-induced emission molecules was up to 598. The chiral recognition could be applied to quantitative analysis of enantiomer content of chiral acids. More chiral AIE amines need to be developed for enantiomer analysis of more carboxylic acids.     

Infinite enantiomeric resolution of basic compounds using highly sulfated cyclodextrin as chiral selector in capillary electrophoresis

Four chiral basic analytes, namely methadone, fluoxetine, venlafaxine, and tramadol, were selected as model compounds for investigating their stereoselective separation with highly sulfated gamma-cyclodextrin (HS gamma-CD) by capillary electrophoresis (CE)-UV and CE-mass spectrometry (MS). At high concentration of chiral selector, the preferentially bonded enantiomer migrated faster in the anodic mode to the detector and high resolutions were obtained for all analytes. In the cathodic mode, at lower highly sulphated cyclodextrin (HS-CD) concentration, basic compounds could be detected, with the weakly bonded enantiomer migrating first (enantiomeric migration order inversion). It was also then possible, at intermediate HS-CD concentration, that only one enantiomer migrated to the detector as cation while the other enantiomer complexed with the CD was negatively charged and presented an opposite mobility. The latter never reached the detector achieving a perfect enantiomeric selectivity. Infinite chiral resolutions were thus achieved by CE-UV as well as by CE-electrospray ionisation (ESI)-MS where concentrations of HS-CD were adapted according to the negative contribution of the nebulization gas pressure of the interface.     

The CGC enantiomer separation of 2-arylcarboxylic acid esters by using β-cyclodextrin derivatives as chiral stationary phases

Chiral 2-arylcarboxylic acid esters are important intermediates in preparation of enantioenriched 2-arylpropionic acids type Non-steroidal anti-inflammatory drugs (NSAIDs). Enantiomer separation of 2-arylcarboxylic acid esters is crucial for evaluation of the asymmetric synthesis efficiency and the enantiomer excess of chiral 2-arylcarboxylic acid derivatives. The capillary gas chromatography (CGC) enantiomer separation of 17 pairs of 2-arylcarboxylic acid esters enantiomers was conducted by using seven different β-cyclodextrin derivatives (CDs) as chiral stationary phases. It was found that for the 7 pairs of 2-phenylpropionates enantiomers, CDs with both alkyl and acyl substituents especially 2,6-di-O-pentyl-3-O-butyryl-β-cyclodextrin exhibited better enantiomer separation abilities than the other CDs examined. For the 7 pairs of 2-(4-substituted phenyl)propionates enantiomers, 2,3,6-tri-O-methyl-β-cyclodextrin possessed better enantiomer separation abilities than the other CDs. Among the 3 pairs of 2-phenylbutyrates enantiomers examined, only methyl 2-phenylbutyrate enantiomers could be separated by three CDs among the 7 CDs tested, while enantiomers of ethyl 2-phenylbutyrate and isopropyl 2-phenylbutyrate couldn't be separated by any of the 7 CDs tested. Besides the structures of CDs, the structures of 2-arylcarboxylic acid esters including different ester moieties, substituents of phenyl, and different carboxylic acids moieties in 2-arylcarboxylic acid esters also affected the enantiomer separation results greatly. The CGC enantiomer separation results of 2-arylcarboxylic acid esters on different CDs are useful for solving the enantiomer separation problem of 2-arylcarboxylic acid esters.     

Asymmetric Synthesis of 2-Arylindolines and 2,2-Disubstituted Indolines by Kinetic Resolution

Kinetic resolution of 2-arylindolines (2,3-dihydroindoles) was achieved by treatment of their N-tert-butoxycarbonyl (Boc) derivatives with n-butyllithium and sparteine in toluene at −78 °C followed by electrophilic quench. The unreacted starting materials together with the 2,2-disubstituted products could be isolated with high enantiomer ratios. Variable temperature NMR spectroscopy showed that the rate of Boc rotation was fast (ΔG^≠≈57 kJ/mol at 195 K). This was corroborated by DFT studies and by in situ ReactIR spectroscopy. The enantioenriched N-Boc-2-arylindolines were converted to 2,2-disubstituted products without significant loss in enantiopurity. Hence, either enantiomer of the 2,2-disubstituted products could be obtained with high selectivity from the same enantiomer of the chiral ligand sparteine (one from the kinetic resolution and the other from subsequent lithiation-trapping of the recovered starting material). Secondary amine products were prepared by removing the Boc group with acid to provide a way to access highly enantioenriched 2-aryl and 2,2-disubstituted indolines.     

Chirally autocatalytic reaction performed in highly supersaturated conditions

Spontaneous chiral asymmetry generation, which is the preferential production of one enantiomer in a non‐chiral environment by chiral autocatalysis, could be observed in a preparation of a octahedral cobalt complex, cis‐[CoBr(NH₃)(en)₂]Br₂. A concentration fluctuation in a far‐from‐equilibrium chemical system will grow if the rate of local autocatalytic production of a compound in a small volume overcomes its loss due to diffusion. In a chirally autocatalytic system, this phenomenon could produce a large variation in the enantiomeric excess. In a reaction that produces the cobalt complex, the reaction rate was found to increase in the highly supersaturated solution of the product. In supersaturated solutions, before crystals of the solute have nucleated, embryos, which are the clusters of the solute, are formed. Ternary water‐solubility isotherm of each enantiomer of the cobalt complex suggests that each embryo consists of one exclusive enantiomer. Each chiral cluster, which could be regarded as polymeric material, thus formed in a highly supersaturated solution, may act as catalyst for the production of the same enantiomer. Life is a far‐from‐equilibrium self‐organized polymeric system in which chiral symmetry is broken. This reaction system is thus a model for the generation and amplification of chiral asymmetry in polymeric materials; it provides some insight in to the mechanisms that might have produced the observed biomolecular homochirality.     

Signal separation and determination of the enantiomeric purity of primary amines with (-)-myrtenal--a 13C NMR study

The applicability of (-)-myrtenal as a chiral derivatizing agent in combination with (13)C NMR spectroscopy was investigated. (13)C NMR was found to be a valuable tool for the identification and enantiomer differentiation of primary amines including beta-amino alcohols and vicinal diamines. The enantiomeric excess could be determined via automated deconvolution and integration, and was found to be in good accordance with the expected values even in the cases, when enantiomer differentiation was not possible in (1)H NMR spectra.     

Enantioselective DNA threading dynamics by phenazine-linked.

The interactions between the stereoisomers of the chiral bis-intercalator [mu-C4(cpdppz)(2)-(phen)(4)Ru(2)](4+) and DNA reveal interesting dynamic discrimination properties. The two enantiomers Delta-Delta and Lambda-Lambda both form very strong complexes with calf thymus DNA with similar thermodynamic affinities. By contrast, they display considerable variations in their binding kinetics. The Delta-Delta enantiomer has higher affinity for calf thymus DNA than for [poly(dA-dT)](2), and the association kinetics of the dimer to DNA, as well as to polynucleotides, requires a multiexponential fitting function. The dissociation reaction, on the other hand, could be described by a single exponential for [poly(dA-dT)](2), whereas two exponentials were required for mixed-sequence DNA. To understand the key mechanistic steps of the reaction, the kinetics was studied at varied salt concentration for different choices of DNA and chirality of the threading complex. The enantiomers were found to have markedly different dissociation rates, the Lambda-Lambda enantiomer dissociating about an order of magnitude faster than the Delta-Delta enantiomer. Also, the salt dependence of the dissociation rate constants differed between the enantiomers, being stronger for the Lambda-Lambda enantiomer than for the Delta-Delta enantiomer. Since the dissociation reaction requires unthreading of bulky parts of the bis-intercalator through the DNA helix, a considerable conformational change of the DNA must be involved, possibly defining the rate-limiting step.     

Light-emitting property of simple AIE compounds in gel, suspension and precipitates, and application to quantitative determination of enantiomer composition

It was found that simple AIE compounds could emit different intensity of fluorescence in gel, suspension and precipitates for the first time, which could be applied to fluorescence switches and quantitative determination of enantiomer composition.     

Molecularly imprinted CEC sorbents: investigations into polymer preparation and electrolyte composition

The influence of the sorbent preparation protocol and separation parameters on the selectivity and chromatographic efficiency of super-porous molecularly imprinted polymer (MIP) monoliths in capillary electrochromatography (CEC) was studied. Chiral templates were employed and enantiomer separation and resolution were used as measures of imprint selectivity and column efficiency, respectively; the latter was in addition studied by chromatography of non-related aromatic structures. The polymer preparation was varied with respect to monomer composition in the pre-polymerisation mixture and also the use of single versus multiple template(s). The separation parameters investigated were type and content of organic solvent and surfactant modifier in the electrolyte. It was found that acetone and acetonitrile in buffer mixtures provided enantiomer separation of enantiomers of the template and also structural analogues; however, the degree of separation was greatly influenced by the content in the electrolyte. Three surfactants, sodium dodecylsulfate (SDS), cetyltrimethylammonium bromide (CTAB) and polyoxyethylene sorbitanmonolaurate (Tween 20), were examined as electrolyte modifiers. It was found that addition of SDS decreased and CTAB and Tween 20 increased the enantiomer separation. SDS and CTAB could be used up to 1 mM concentration whereas Tween could be used up to 90 mM concentration without causing baseline disturbances. The effects found and demonstrated strongly suggest that these parameters are to be considered during optimisation of an MIP-CEC system.     

Enantioselective transesterification catalysis by nanosized serine protease subtilisin Carlsberg particles in tetrahydrofuran

Enzyme catalysis in organic solvents is a powerful tool for stereo-selective synthesis but the enantioselectivity is still hard to predict. To overcome this obstacle, we employed a nanoparticulate formulation of subtilisin Carlsberg (SC) and designed a series of 14 structurally related racemic alcohols. They were employed in the model transesterification reaction with vinyl butyrate and the enantioselectivities were determined. In general, short alcohol side chains led to low enantioselectivties, while larger and bulky side chains caused better discrimination of the enantiomers by the enzyme. With several bulky substrates high enantioselectivities with E>100 were obtained. Computational modeling highlighted that key to high enantioselectivity is the discrimination of the R and S substrates by the sole hydrophobic binding pocket based on their size and bulkiness. While bulky S enantiomer side chains could be accommodated within the binding pocket, bulky R enantiomer side chains could not. However, when also the S enantiomer side chain becomes too large and does not fit into the binding pocket anymore, enantioselectivity accordingly drops.     

Enantioselective separation of some polyunsaturated epoxy fatty acids by high-performance liquid chromatography on a cellulose phenylcarbamate (Chiralcel OC) stationary phase.

cis-Epoxides of linoleic acid, alpha-linolenic acid, arachidonic acid and the omega 3-epoxide of eicosapentaenoic acid were chromatographed on a cellulose trisphenylcarbamate (Chiralcel OC) stationary phase in the normal-phase mode. The R,S and S,R enantiomers of methyl-14(15)epoxyeicosatrienoate, methyl-9(10)epoxyoctadecadienoate and methyl-9(10)epoxyoctadecenoate could be partly resolved. The R,S enantiomer of methyl-14(15)epoxyeicosatrienoate eluted before the S,R enantiomer. [14C]14(15)Epoxyeicosatrienoic acid was isolated from an incubation of [14C]20:4n-6 with microsomes of rabbit kidney cortex and the S,R enantiomer was found to predominate (about 2:1).     

Resolution and stereochemistry of tert-butylphenylphosphinous acid-borane

A combined use of ephedrine and cinchonine as resolving agents enabled facile resolution of racemic tert-butylphenylphosphinous acid-borane (1) into the two enantiomers in ca. 31-32% yield each. The resolved 1 served as a model substrate to study stereoselective synthetic transformations of phosphinous acid-boranes yielding optically active phosphinite-borane, boranatophosphinous-sulfonic anhydride, secondary phosphine-borane, tertiary phosphine-borane, secondary phosphine oxide, and phosphinic halides. By the judicious choice of the reaction paths, either enantiomer of tert-butylphenylphosphine-borane and of tert-butylmethylphenylphosphine-borane could be stereoselectively obtained from a single enantiomer of 1.     

Preparative LC separation and isolation of enantiomerically pure olefins

Summary Diastereomer formation by the reaction of chiral olefins with optically active platinum complexes followed by LC separation of the diastereomers, and on-line or off-line release of the olefins from the complexes by ligand exchange yield optically pure enantiomers.The 100% dextrostetatory enantiomer of exo-2-vinylbicyclo[2.2.1]heptane (vinylnorbornane) could be isolated by an off-line method and was used for polarimetric measurements before and after preparative scale gas chromatographic clean-up.The initial valuable platinum complex could be easily recovered without loss.     

Chiral synthesis of the ABC-ring system of quinocarcin

Stereoselective synthesis of an enantiomeric pair of the ABC-ring system of quinocarcin(1), a notable antitumor antibiotic, could be achieved in> 95%ee by utilizing each enantiomer of 4-O-benzyl-2,3-O-isopropylidene-threose as a chiral auxiliary and featuring novel diastereoselective reduction of the 1,3-disubstituted isoquinoline as a key step.     

Stereoselective and Asymmetric-Selective Polymerization of α-Amino Acid N-Carboxyanhydride

The enantiomer selectivity in the propagation reaction of NCA was investigated by using suitable model reactions. Contrary to the assumption usually made, the enantiomer selectivity in the nucleophilic addition of chiral amines to NCA depended strongly on the structure of amine or NCA and the solvent. In the polymerization by an activated-NCA mechanism, the addition of activated NCA to NCA was found for the first time to be enantiomer-selective. In addition to this, the chiral penultimate unit was found to participate in the enantiomer selection. Structures of the transition states leading to the different types of enantiomer selection were proposed.     

Use of peak sharpening effects to improve the separation of chiral compounds with molecularly imprinted porous polymer layer open‐tubular capillaries

This investigation demonstrates the application of a new peak sharpening technique to improve the separation of difficult‐to‐resolve racemic mixtures in capillary electro‐chromatography. Molecularly imprinted porous layer open tubular (MIP‐PLOT) capillaries, prepared by a layer‐on‐layer polymerization approach with Z‐l ‐Asp‐OH as the template, were selected to validate the approach. SEM revealed that the polymer film thickness can be varied by changes in both the polymer composition and the layer‐on‐layer regime. Capillaries made with methacrylic acid as the functional monomer could not separate the Z‐Asp‐OH racemate, due to weak interactions between the MIP‐PLOT material and the target analytes. In contrast, MIP‐PLOT capillaries prepared with 4‐vinylpyridine as the functional monomer resulted in increased ionic interactions with the target analytes. Separation of the enantiomers could be enhanced when a peak zone sharpening effect was exploited through the use of specific BGE compositions and by taking advantage of eigenpeak phenomena. In this manner, the position of a sharpening zone and the peak shape of the sample analytes could be fine‐tuned, so that when the sharpening zone and the target analyte co‐migrated the separation of the Z‐l ‐Asp‐OH enantiomer from its d ‐enantiomer in a racemic mixture could be achieved under overloading conditions.     

Design criteria for dense permeation-selective membranes for enantiomer separations

Dense enantioselective membranes can distinguish between two enantiomers by different mechanisms. At this moment, it is not clear which mechanism provides the best membranes for large-scale enantiomer separations. Therefore, we studied the design criteria for permeation-selective membranes combining literature data, experiments and model calculations. Literature data on dense permeation-selective membranes for enantiomer separation show that these membranes could be divided into two different classes: diffusion selective and sorption selective. Reviewing the literature on diffusion-selective membranes shows that these membranes have one main disadvantage: the inverse proportionality relation between the permeability and selectivity. This disadvantage is absent for sorption-selective membranes. As a model system, the diffusion of phenylalanine through a packed bed of polypropylene beads coated with N-dodecyl-l-hydroxyproline:Cu(II) was studied. The experiments showed that the material could selectively adsorb phenylalanine (Phe) with a selectivity (d/l) of 1.25. However, no permeation selectivity could be detected. With a dual sorption model these results could be interpreted. These model calculations showed that the permeation selectivity only approaches the intrinsic selectivity of the selector if the selectively adsorbed population is mobile and the non-selective permeation is minimized. Therefore, to our opinion more emphasis should be put on the development of sorption selective membranes.