Chiral sensing of Eu(III)‐containing achiral polymer complex from chiral amino acids coordination induction

The β‐diketonate‐based achiral polymer P‐1 could be synthesized by the polymerization of 3,7‐dibromo‐2,8‐dimethoxy‐5,5‐dioctyl‐5H‐dibenzo[b,d]silole ( M1 ) with (Z)?1,3‐bis(4‐ethynylphenyl)?3‐hydroxyprop‐en‐1‐one ( M2 ) via typical Sonogashira coupling reaction. The β‐diketonate unit in the main chain backbone of P‐1 can further coordinate with Eu(TTA)_x [TTA^? = 4,4,4‐trifluoro‐1‐(thiophen‐2‐yl)butane‐1,3‐dionate anion, X = 1, 2, 3] to afford corresponding Eu(III)‐containing polymer complexes. The resulting achiral polymer complex P‐2 (X = 2) can exhibit strong circular dichroism (CD) response toward both N‐Boc‐l and d‐ proline enantiomers. The CD signal was preliminarily attributed to coordination induction between chiral N‐Boc‐proline and the Eu(III) complex moiety. The linear regression analysis of CD sensing shows a good agreement between the magnitude of molar ellipticity and concentration of chiral N‐Boc‐l or d‐ proline, which indicates this kind Eu(III)‐containing achiral polymer complex can be used as a chiral probe for enantioselective recognition of N‐Boc‐l or d‐ proline enantiomers based on Cotton effect of CD spectra. © 2014 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2014 , 52, 3080–3086     

Preparative enantioseparation of propafenone by counter‐current chromatography using di‐n‐butyl l‐tartrate combined with boric acid as the chiral selector

This paper extends the research of the utilization of borate coordination complexes in chiral separation by counter‐current chromatography (CCC). Racemic propafenone was successfully enantioseparated by CCC with di‐n‐butyl l ‐tartrate combined with boric acid as the chiral selector. The two‐phase solvent system was composed of chloroform/ 0.05 mol/L acetate buffer pH 3.4 containing 0.10 mol/L boric acid (1:1, v/v), in which 0.10 mol/L di‐n‐butyl l ‐tartrate was added in the organic phase. The influence of factors in the enantioseparation of propafenone were investigated and optimized. A total of 92 mg of racemic propafenone was completely enantioseparated using high‐speed CCC in a single run, yielding 40–42 mg of (R)‐ and (S)‐propafenone enantiomers with an HPLC purity over 90–95%. The recovery for propafenone enantiomers from fractions of CCC was in the range of 85–90%.     

Stereoselective determination of dufulin in watermelon under field conditions using chiral ultra high performance liquid chromatography with high‐resolution mass spectrometry

An effective and sensitive chiral analytical method was established to investigate the stereoselective dissipation of rac‐dufulin in watermelon using ultra high performance liquid chromatography with a superchiral S‐OD chiral column (4.6 × 150 mm i.d., 5 μm) coupled with high‐resolution mass spectrometry. To optimize the pretreatment method for detecting rac‐dufulin in the three matrixes, different extraction solvents, extractant volumes, extraction times, and absorbents were investigated to improve extraction efficiency. Moreover, analysis of variance was used to perform method validation for determination of the two dufulin enantiomers in the three matrixes. Using the optimized method, good linearity was obtained (determination coefficient > 0.999). The limits of detection and quantification of the two dufulin enantiomers in soil, watermelon, and pulp were 0.15 and 0.5 μg/kg, respectively. The average recoveries of the two enantiomers in the three matrixes at four spiked levels ranged from 75.0 to 107.8%, with intra‐ and inter‐day relative standard deviations of 0.4–10.4%. In field trials, the R enantiomer was preferentially dissipated in watermelon. These method validation results confirmed that the developed method was convenient and reliable for the stereoselective determination of enantiomers of rac‐dufulin in watermelon.     

Indirect reversed‐phase high‐performance liquid chromatographic and direct thin‐layer chromatographic enantioresolution of (R,S)‐Cinacalcet

Enantioresolution of the calcimimetic drug (R,S)‐Cinacalcet was achieved using both indirect and direct approaches. Six chiral variants of Marfey's reagent having l ‐Ala‐NH₂, l ‐Phe‐NH₂, l ‐Val‐NH₂, l ‐Leu‐NH₂, l ‐Met‐NH₂ and d ‐Phg‐NH₂ as chiral auxiliaries were used as derivatizing reagents under microwave irradiation. Derivatization conditions were optimized. Reversed‐phase high‐performance liquid chromatography was successful using binary mixtures of aqueous trifluoroacetic acid and acetonitrile for separation of diastereomeric pairs with detection at 340 nm. Thin silica gel layers impregnated with optically pure l ‐histidine and l ‐arginine were used for direct resolution of enantiomers. The limit of detection was found to be 60 pmol in HPLC while in TLC it was found to be in the range of 0.26–0.28 µg for each enantiomers. Copyright © 2010 John Wiley & Sons, Ltd.     

Sensitive determination of D-lactic acid and L-lactic acid in urine by high-performance liquid chromatography-tandem mass spectrometry

d ‐Lactic acid in urine originates mainly from bacterial production in the intestinal tract. Increased d ‐lactate excretion as observed in patients affected by short bowel syndrome or necrotizing enterocolitis reflects d ‐lactic overproduction. Therefore, there is a need for a reliable and sensitive method able to detect d ‐lactic acid even at subclinical elevation levels. A new and highly sensitive method for the simultaneous determination of l ‐ and d ‐lactic acid by a two‐step procedure has been developed. This method is based on the concentration of lactic acid enantiomers from urine by supported liquid extraction followed by high‐performance liquid chromatography–tandem mass spectrometry. The separation was achieved by the use of an Astec Chirobiotic? R chiral column under isocratic conditions. The calibration curves were linear over the ranges of 2–400 and 0.5–100 µmol/L respectively for l ‐ and d ‐lactic acid. The limit of detection of d ‐lactic acid was 0.125 µmol/L and its limit of quantification was 0.5 µmol/L. The overall accuracy and precision were well within 10% of the nominal values. The developed method is suitable for production of reference values in children and could be applied for accurate routine analysis. Copyright © 2011 John Wiley & Sons, Ltd.     

Helix formation of poly(phenylacetylene) derivatives bearing amino groups at the meta position induced by optically active carboxylic acids

Two novel phenylacetylene derivatives bearing diethylaminomethyl groups at the meta position on phenyl groups [3‐(N,N‐diethylaminomethyl)phenyl]acetylene ( 1 ) and [3,5‐bis(N,N‐diethylaminomethyl)phenyl]acetylene ( 2 ) were synthesized and polymerized with [Rh(nbd)Cl]₂ (nbd: norbornadiene). Both monomers gave highly cis–transoidal stereoregular polymers that exhibited an induced circular dichroism (ICD) in the UV–visible region, probably because of a prevailing one‐handed helical conformation upon complexation with optically active carboxylic acids such as mandelic acid and lactic acid. The sign of the Cotton effects reflected the absolute configuration of the chiral acids. Therefore, these polymers can be used as a novel probe for determining the configuration of chiral acids. The polymers were stable in the presence of chiral acids in solution. The poly‐ 1 complexed with chiral acids exhibited a split‐type ICD, whereas the poly‐ 2 complexed with chiral acids showed a different, non‐split‐type ICD. The ICD pattern of the poly‐ 1 /chiral acids complexes dramatically changed with an increase in the concentration of the chiral acids, thus showing a non‐split‐type ICD similar to those of the poly‐ 2 /chiral acid complexes. © 2001 John Wiley & Sons, Inc. J Polym Sci Part A: Polym Chem 39: 3180–3189, 2001     

Determination of the enantiomers of α‐hydroxy‐ and α‐amino acids in capillary electrophoresis with contactless conductivity detection

The enantiomers of the anions of five α‐hydroxy acids, namely lactic acid, α‐hydroxybutyric acid, 2‐hydroxycaproic acid, 2‐hydroxyoctanoic acid and 2‐hydroxydecanoic acid, as well as the two α‐amino acids aspartic acid and glutamic acid, were baseline separated and detected by CE with contactless conductivity detection. Vancomycin was employed as chiral selector and could be used with conductivity detection without having to resort to a partial filling protocol as needed when this reagent is used with UV absorbance measurements. The procedure was successfully applied to the determination of the lactic acid enantiomers in samples of milk and yogurt. Linearity was achieved in the concentration range of 10–500 μmol/L with good correlation coefficients (0.9993 and 0.9990 for L ‐ and D ‐lactic acid, respectively). The LODs (3 S/N) for L ‐ and D ‐lactic acid were determined as 2.8 and 2.4 μmol/L, respectively.     

Natural terpene derivatives as new structural task‐specific ionic liquids to enhance the enantiorecognition of acidic enantiomers on teicoplanin‐based stationary phase by high‐performance liquid chromatography

We present the specific cooperative effect of a semisynthetic glycopeptide antibiotic teicoplanin and chiral ionic liquids containing the (1R ,2S ,5R )‐(–)‐menthol moiety on the chiral recognition of enantiomers of mandelic acid, vanilmandelic acid, and phenyllactic acid. Experiments were performed chromatographically on an Astec Chirobiotic T chiral stationary phase applying the mobile phase with the addition of the chiral ionic liquids. The stereoselective binding of enantiomers to teicoplanin in presence of new chiral ionic liquids were evaluated applying thermodynamic measurements and the docking simulations. Both the experimental and theoretical methods revealed that the chiral recognition of enantiomers in the presence of new chiral ionic liquids was enthalpy driven. The changes of the teicoplanin conformation occurring upon binding of the chiral ionic liquids are responsible for the differences in the standard changes in Gibbs energy (ΔG ⁰) values obtained for complexes formed by the R and S enantiomers and teicoplanin. Docking simulations revealed the steric adjustment between the chiral ionic liquids cyclohexane ring (chair conformation) and the β‐d ‐glucosamine ring of teicoplanin and additionally hydrophobic interactions between the decanoic aliphatic chain of teicoplanin and the alkyl group of the tested salts. The obtained terpene derivatives can be considered as “structural task‐specific ionic liquids” responsible for enhancing the chiral resolution in synergistic systems with two chiral selectors.     

Enantiomeric separation of d‐ and l‐serine using high‐performance liquid chromatography with electrochemical detection following pre‐column diastereomer derivatization

Enantiomeric separation of d ‐ and l ‐serine on an octadecylsilica column was investigated using (2R)‐2,5‐dioxopyrrolidin‐1‐yl‐2,5,7,8‐tetramethyl‐6‐(tetrahydro‐2H‐pyran‐2‐yloxy)chroman‐2‐carboxylate (R‐NPCA), which was developed for a pre‐column derivatization reagent for electrochemical detection. In addition, (2S)‐2,5‐dioxopyrrolidin‐1‐yl‐2,5,7,8‐tetramethyl‐6‐(tetrahydro‐2H‐pyran‐2‐yloxy)chroman‐2‐carboxylate (S‐NPCA) was newly synthesized from (S)‐(?)‐6‐hydroxy‐2,5,7,8‐tetramethylchroman‐2‐carboxylic acid (S‐α‐CA), and the enantiomeric separation of d ‐ and l ‐serine using S‐NPCA was also examined. The enantiomeric separation of d ,l ‐serine was achieved using the R‐ or S‐NPCA as a chiral derivatization reagent, and the elution orders of the enantiomers were reversed between R‐ and S‐NPCA. The elution orders of d ‐ and l ‐serine unexpectedly reversed between the phosphate buffer at pH 4.0 and pH 2.2, both of which were used in the mobile phase. Separation factors obtained using R‐ and S‐NPCA were similar—1.09 and 1.07, respectively. The detection limit was approximately 940 fmol on the column (signal‐to‐noise ratio 3) when the applied voltage was +650 mV. Copyright © 2009 John Wiley & Sons, Ltd.     

Chiral separation of disease biomarkers with 2‐hydroxycarboxylic acid structure

Chiral 2‐hydroxycarboxylic acids are compounds that have been linked to particular diseases and are putative biomarkers with some diagnostic potential. The importance of identifying whether a particular enantiomer is related to certain diseases has been encouraged recently. However, in many cases it has not yet been elucidated whether there are stereochemical implications with respect to these biomarkers and whether their enantioselective analysis provides new insights and diagnostic potential. In this study 13 disease‐related chiral 2‐hydrocarboxylic acids were studied for their chiral separation by high‐performance liquid chromatography on three cinchona alkaloid‐derived chiral stationary phases. From a subgroup of eight 2‐hydroxymonocarboxylic acids, baseline resolution could be achieved and inversion of elution order by exchanging tert‐butylcarbamoyl quinidine chiral stationary phase (Chiralpak QD‐AX) for the corresponding quinine analogue (Chiralpak QN‐AX) is shown for seven of them. Furthermore, conditions for chiral separation of the 2‐hydroxydicarboxylic acids, citramalic acid, 2‐isopropylmalic acid, and 2‐hydroxyadipic acid are reported and compared to the previous reported conditions for 2‐hydroxyglutaric acid and malic acid.     

Use of chiral derivatization for the determination of dichlorprop in tea samples by ultra performance LC with fluorescence detection

Dichlorprop is available for agricultural use as a chiral pesticide. In this study, the stereoselective determination of dichlorprop enantiomers in tea samples such as green, black, jasmine, and oolong was developed by ultra performance LC with fluorescence spectrometry after covalent chiral derivatization. The separation was achieved on an Acquity BEH C18 column with the mobile phase consisting of 0.1% formic acid in acetonitrile/water at a flow rate of 0.4 mL/min. In the covalent chiral derivatization using (S)‐(+)‐4‐(N,N‐dimethylaminosulfonyl)‐7‐(3‐aminopyrrolidin‐1‐yl)‐2,1,3‐benzoxadiazole, the peak resolution between the S and R‐dichlorprop enantiomers was 2.6. LODs and LOQs values were 10 and 50 ng/mL standard solution. The linearity of the calibration curves yielded the coefficients (r² > 0.99, ranging from 0.05 to 5 μg/mL) of determination of each of the dichlorprop enantiomers. SPE extraction was used for the sample preparation of dichlorprop in various tea samples. Recoveries were in the range of 82.4–97.6% with associated precision values (within‐day: 82.4–95.8%, n = 6, and between‐day: 83.7–97.6% for 3 days) for repeatability and reproducibility. Based on this result, our method has been proven to be highly efficient and suitable for the routine assay of dichlorprop enantiomers in various tea samples. We propose that the ultra performance LC assay after covalent chiral derivatization would be the renewed tools in the era of chiral stationary platform for chiral pesticide residues in foods.     

Preparative enantioseparation of loxoprofen precursor by recycling countercurrent chromatography with hydroxypropyl‐β‐cyclodextrin as a chiral selector

Recycling countercurrent chromatography was successfully applied to the resolution of 2‐(4‐bromomethylphenyl)propionic acid, a key synthetic intermediate for synthesis of nonsteroidal anti‐inflammatory drug loxoprofen, using hydroxypropyl‐β‐cyclodextrin as chiral selector. The two‐phase solvent system composed of n‐hexane/n‐butyl acetate/0.1 mol/L citrate buffer solution with pH 2.4 (8:2:10, v/v/v) was selected. Influence factors for the enantioseparation were optimized, including type of substituted β‐cyclodextrin, concentration of hydroxypropyl‐β‐cyclodextrin, separation temperature, and pH of aqueous phase. Under optimized separation conditions, 50 mg of 2‐(4‐bromomethylphenyl)propionic acid was enantioseparated using preparative recycling countercurrent chromatography. Technical details for recycling elution mode were discussed. The purities of both the S and R enantiomers were over 99.0% as determined by high‐performance liquid chromatography. The enantiomeric excess of the S and R enantiomers reached 98.0%. The recovery of the enantiomers from eluted fractions was 40.8–65.6%, yielding 16.4 mg of the S enantiomer and 10.2 mg of the R enantiomer. At the same time, we attempted to enantioseparate the anti‐inflammatory drug loxoprofen by countercurrent chromatography and high‐performance liquid chromatography using a chiral mobile phase additive. However, no successful enantioseparation was achieved so far.     

Binding modes identification through molecular dynamic simulations: A case study with carnosine enantiomers and the Teicoplanin A2‐2‐based chiral stationary phase

In the present study, an in silico methodology able to define the binding modes adopted by carnosine enantiomers in the setting of the chiral recognition process is described. The inter‐ and intramolecular forces involved in the enantioseparation process with the Teicoplanin A2‐2 chiral selector and carnosine as model compound are successfully identified. This approach fully rationalizes, at a molecular level, the (S) < (R) enantiomeric elution order obtained under reversed‐phase conditions. Consistent explanations were achieved by managing molecular dynamics results with advanced techniques of data analysis. As a result, the time‐dependent identification of all the interactions simultaneously occurring in the chiral selector‐enantiomeric analyte binding process was obtained. Accordingly, it was found that only (R)‐carnosine is able to engage a stabilizing charge–charge interaction through its ionized imidazole ring with the carboxylate counter‐part on the chiral selector. Instead, (S)‐carnosine establishes intramolecular contacts between its ionized functional groups, that limit its conformational freedom and impair the association with the chiral selector unit.     

Polymer‐based monolithic column with incorporated chiral metal–organic framework for enantioseparation of methyl phenyl sulfoxide using nano‐liquid chromatography

A new approach to the preparation of enantioselective porous polymer monolithic columns with incorporated chiral metal–organic framework for nano‐liquid chromatography has been developed. While no enantioseparation was achieved with monolithic poly(4‐vinylpyridine‐co‐ethylene dimethacrylate) column, excellent separations of both enantiomers of (±)‐methyl phenyl sulfoxide were achieved with its counterpart prepared after admixing metal–organic framework [Zn₂(benzene dicarboxylate)(l‐lactic acid)(dmf)], which is synthesized from zinc nitrate, l ‐lactic acid, and benzene dicarboxylic acid in the polymerization mixture. These novel monolithic columns combined selectivity of the chiral framework with the excellent hydrodynamic properties of polymer monoliths, may provide a great impact on future studies in the field of chiral analysis by liquid chromatography.     

One‐dimensional TOCSY 1H NMR experiments on adducts of amino acid esters with cobalt(III) tetramethylchiroporphyrin. Prospects for the analysis of amino acid mixtures

Cobalt(III) tetramethylchiroporphyrin, CoCl(TMCP), is a useful chiral shift reagent for structure attribution, absolute configuration assignment and enantiomeric excess determination of amino acid methyl esters by ¹H NMR spectroscopy. However, it has two axial sites available for amine coordination, a structural feature which generates n(n + 1)/2 diastereomeric species and n² distinct spin systems from a mixture of n amino ester enantiomers, making the analysis of complex amino acid samples exceedingly difficult by classical 1‐D or 2‐D NMR methods when n > 3. The 1‐D TOCSY experiment is shown to be a powerful tool for the selective excitation and detection of every single component of a mixture of four amino acid methyl esters bound to CoCl(TMCP): those of(S)‐Leu, (S)‐Asp, (R)‐Asp and (S)‐Glu, for example. The potential utility of this methodology for the determination of amino acid enantiomers in carbonaceous meteorites or other extraterrestrial samples is suggested. Copyright © 2002 John Wiley & Sons, Ltd.     

Enantioseparation of aromatic amino acids using CEC monolith with novel chiral selector,N‐methacryloyl‐l‐histidine methyl ester

A new type of polymethacrylate‐based monolithic column with chiral stationary phase was prepared for the enantioseparation of aromatic amino acids, namely d ,l ‐phenylalanine, d ,l ‐tyrosine, and d ,l ‐tryptophan by CEC. The monolithic column was prepared by in situ polymerization of butyl methacrylate (BMA), N‐methacryloyl‐l ‐histidine methyl ester (MAH), and ethylene dimethacrylate (EDMA) in the presence of porogens. The porogen mixture included DMF and phosphate buffer. MAH was used as a chiral selector. FTIR spectrum of the polymethacrylate‐based monolith showed that MAH was incorporated into the polymeric structure via in situ polymerization. Some experimental parameters including pH, concentration of the mobile phase, and MAH concentration with regard to the chiral CEC separation were investigated. Single enantiomers and enantiomer mixtures of the amino acids were separately injected into the monolithic column. It was observed that l ‐enantiomers of aromatic amino acids migrated before d ‐enantiomers. The reversal enantiomer migration order for tryptophan was observed upon changing of pH. Using the chiral monolithic column (100 μm id and 375 μm od), the best chiral separation was performed in 35:65% ACN/phosphate buffer (pH 8.0, 10 mM) with an applied voltage of 12 kV in CEC. SEM images showed that the chiral monolithic column has a continuous polymeric skeleton and large through‐pore structure.     

Enantioselective determination of ketamine in dog plasma by chiral liquid chromatography–tandem mass spectrometry

Ketamine is an N‐methyl‐d ‐aspartate receptor antagonist that is usually used clinically as a racemic mixture. Its two enantiomers exhibit different pharmacological activities. To determine whether the enantiomers have different pharmacokinetic profiles, a chiral liquid chromatography–tandem mass spectrometry method was developed and validated for the determination of ketamine enantiomers in dog plasma. The enantiomers of ketamine were extracted from 50 μL of plasma by methyl tert‐butyl ether. Adequate chromatographic retention and baseline resolution of the enantiomers were achieved within a runtime of 5 min on a chiral column coated with polysaccharide derivatives, using a gradient mobile phase of acetonitrile and 10 mm ammonium bicarbonate aqueous solution. Ketamine enantiomers were detected by mass spectrometry with multiple reaction monitoring mode using the transitions of m/z 238.3 → 125.9 for the analytes and m/z 237.1 → 194.1 for carbamazepine (internal standard). The method was linear over the concentration range from 0.5 to 500 ng/mL for each enantiomer. The lower limit of quantification (LLOQ) for each enantiomer was 0.5 ng/mL. The intra‐ and inter‐day precision was <7.3% and 8.5% for R‐ and S‐ketamine, respectively. The accuracy was 92.9–110.4% for R‐ketamine and 99.8–102.4% for S‐ketamine. The method was successfully applied to characterize the stereoselective pharmacokinetic profiles of ketamine in beagle dogs.     

Partial separation of lactic acid enantiomers by HPLC using (S)-(+)mandelic acid as chiral mobile-phase additive

Summary Separation of lactic acid enantiomers by inductive adsorption has been studied on silica columns using (S)-(+)mandelic acid as the chiral mobile-phase additive. The lactic acid peak is not very strongly retained, broad, strongly tailing and splits in two at loadings lower than 60 mol cm^–2 column cross-section and at mandelic acid concentrations higher than 10^–4M. The splitting indicates partial separation of enantiomers: enantiomerically enriched fractions are obtained but not enantiomerically pure. Lactic acid retention strongly decreases with increasing concentration of both hydroxy-acids; mandelic acid acts by displacement and lactic acid acts by an overloading effect.     

Enantioselective analysis of venlafaxine and its active metabolites: A review on the separation methodologies

Venlafaxine (VFX) is a serotonin and norepinephrine reuptake inhibitor chiral drug used in therapy as an antidepressant in the form of a racemate consisting of R‐ and S‐VFX. The two enantiomers of VFX exhibit different pharmacological activities: R‐VFX inhibits both norepinephrine and serotonin synaptic reuptake, whereas S‐VFX inhibits only the serotonin one. R‐ and S‐VFX are metabolized in the liver to the respective R‐ and S‐O‐desmethylvenlafaxine (ODVFX), R‐ and S‐N‐desmethylvenlafaxine (NDVFX), and R‐ and S‐N,O‐didesmethylvenlafaxine (NODVFX). The pharmacological profile of ODVFX is close to that of VFX, whereas the other two chiral metabolites (NDVFX and NODVFX) have lower affinity for the receptor sites. The pharmacokinetics of the VFX enantiomers appear stereoselective, including the metabolism process. In the past 20 years, several studies describing the enantioselective analysis of R‐ and S‐VFX in pharmaceutical formulations and its chiral metabolites in biological matrices were published. These methods encompass liquid chromatography coupled with UV detection, mass spectrometry, or tandem mass spectrometry, and capillary electrophoresis. This paper reviews the published methods used for the determination of the individual enantiomers of VFX and its chiral metabolites in different matrices.     

Modified Cyclodextrins as Enantioselective Hosts for Amino Acids

Two modified β‐cyclodextrins, H‐2 and H‐3 , having a flexible appended moiety were studied for the chiral discrimination of the enantiomers of various amino acids by means of fluorescence as signaling option. These hosts quenched the fluorescence intensities of amino acids upon binding. The d‐ enantiomers were better recognized by these hosts. The association constants (K_s) and enantioselectivity factors (α) of the host?guest complexes were calculated.