Enantioseparation of Phenylsuccinic Acid Enantiomers by Solvent Sublation with Collaborative Selectors

A new solvent sublation (SS) system for chiral separation is introduced by using phenylsuccinic acid (H₂A) as the model enantiomers. The experiments were carried out in a traditional SS apparatus but with collaborative chiral selectors: dibenzoyl-L-tartaric acid (L-DBTA) in the organic phase and hydroxypropyl-β-cyclodextrin (HP-β-CD) in the aqueous phase. The chiral recognition abilities of the two selectors are opposite for the H₂A enantiomers. Several important parameters were investigated. The results demonstrate that enantioselective sublation and partitioning behavior are mainly dependent on the pH of the solution, the concentrations of chiral selectors and H₂A. Furthermore, the flow rate of air and flotation time also have some effects on the enantioseparation. Under the optimized conditions, the enantioselectivity expressed by the separation factor (β) and enantiomer excess (e.e.%) are 2.47 and 29.50%, and the yields of R-H₂A and S-H₂A are 0.23 and 0.13 g·L^?1, respectively. Compared with the SS system with the single selector HP-β-CD in the aqueous phase (or L-DBTA in the organic phase), the increased values of β and e.e.% in the new SS system with collaborative selectors are 1.31 (or 1.38) and 5.90% (or 13.82%), respectively.     

Chiral separation of α-cyclohexylmandelic acid enantiomers using ionic liquid/salt aqueous two-phase system

In this paper, an ionic liquid-based aqueous two-phase system (ILATPS) was applied to the chiral separation of α-cyclohexylmandelic acid (α-CHMA) enantiomers with hydroxypropyl-β-cyclodextrin (HP-β-CD) as the chiral selector. Several influencing parameters were investigated including the types and concentration of ionic liquids, the amount of phase-forming salt, temperature, mixing time, pH, and the content of HP-β-CD. The results showed that not all ILATPS had the ability to chirally recognise the selected enantiomers and that [C₄mim]BF₄/(NH₄)₂SO₄-based ATPS possessed the best enantioseparation ability of the investigated ILATPSs. Under optimal conditions, the separation factor (α) attained 1.59 in a single-step extraction. ILATPS is much “greener” than other liquid-liquid extraction systems, showing its potential for application to the chiral separation.     

LC Enantioseparation of Aryl-Substituted β-Lactams Using Variable-Temperature Conditions

Direct reversed-phase high-performance liquid chromatographic methods were developed for the separation of enantiomers of β-lactams. The enantiomers of 7 aryl-substituted β-lactams were separated on chiral stationary phases containing the macrocyclic glycopeptide antibiotic teicoplanin (Chirobiotic T) and teicoplanin aglycone (Chirobiotic TAG) at 10-°C increments in the range 5–45 °C, using different compositions of 0.1% aqueous triethylammonium acetate (pH 4.1)/methanol (v/v) as mobile phase. The mobile phase composition and temperature were varied to achieve baseline resolutions in a single chromatographic run. The dependence of the natural logarithms of the selectivity factors ln α on the inverse of temperature, 1/T, was used to determine the thermodynamic data on the enantiomers. The thermodynamic data revealed that all the compounds in this study undergo separation via the same enthalpy-driven chiral recognition mechanism. The different methods were compared in systematic chromatographic examinations. The effects of the organic modifier, the mobile phase composition and the temperature on the separation were investigated.     

Enantioseparation of Hydrobenzoin and Structurally Related Compounds on <Emphasis Type="Italic">β</Emphasis>-Cyclodextrin and Hydroxypropyl-<Emphasis Type="Italic">β</Emphasis>-cyclodextrin Bonded Chiral Stationary Phases

A β-cyclodextrin (β-CD) and a hydroxypropyl-β-cyclodextrin (HP-β-CD) bonded chiral stationary phase (CSP) were prepared. Comparative evaluations of these two CSPs for the enantioseparation of hydrobenzoin, benzoin and α-phenethyl alcohol by reversed-phase liquid chromatography were presented. The effects of buffer composition in the mobile phase on the retention and enantioseparation were investigated. The borate buffer had a significant influence on the retention and enantioseparation of hydrobenzoin. Linear solvent strength retention model was used to fit the chromatographic data. Good linearity existed between the logarithm of retention factor (k) and the volume fraction of organic modifier (φ). Another retention model, stoichiometric displacement theory for retention, was also tried to fit the chromatographic data. The results showed that not only acetonitrile, but also water molecules participated in the displacing process of the solute.     

Preparation and characterization of the inclusion complexes of equol with sulfobutylether-β-cyclodextrin: their antioxidant activity and dissolution evaluation

The formation of inclusion complexes between S-(?)-equol (SEq) and cyclodextrins (CDs) was investigated. The binding constant (K_c) of the SEq/sulfobutylether-β-cyclodextrin (SBE-β-CD) inclusion complex was determined to be 1600 L/mol based on UV data. The phenyl ring of the SEq molecule was found to be inserted from the secondary hydroxyl face of the SBE-β-CD as evidenced from ¹H–¹H rotating frame nuclear Overhauser effect spectroscopy (ROESY) NMR. The thermal properties of the solid SEq/SBE-β-CD inclusion complexes prepared by physical mixing, kneading and freeze-drying methods were studied by differential scanning calorimetry. For the solid complex obtained by the freeze-drying method, the endothermic peak corresponding to the melting point of SEq disappeared. The solid SEq/SBE-β-CD complexes exhibited a high score in antioxidant activity evaluation tests compared to SEq alone. Dissolution test revealed that the solid complex obtained by freeze-drying method had improved dissolution of SEq.     

Determination of the enantiomeric purity of albuterol on sorbents modified by macrocyclic antibiotics

The separation of albuterol enantiomers on sorbents with macrocyclic glycopeptide antibiotics immobilized on the silica surface was investigated. Commercial columns—Nautilus-E (BioKhimMak, Russia) with eremomycin as a chiral selector and ChirobioticTAG (Astec, United States) with teicoplanin aglycone as a chiral selector—were used for enantioseparation. Levalbuterol is the (R)-enantiomer of albuterol. We managed to separate albuterol enantiomers on both columns in a polar organic mode, but selectivity was higher on the ChirobioticTAG column (R _s = 1.7). The maximum resolution of enantiomer peaks (1.7) was observed in methanol–acetonitrile–triethylamine–acetic acid (90: 10: 0.05: 0.05) as the mobile phase. The detection limit of the compound calculated by a signal–background ratio of 3: 1 was 0.00002 mg/mL, which corresponds to 0.1% of (S)-enantiomer with respect to the total amount. The results made it possible to determine the enantiomeric purity of the active pharmaceutical substances of levalbuterol.     

The Role of Temperature in Enantioseparation of Norgestrel with Native Cyclodextrins: A Combined LC and NMR Study

Temperature dependence (10–50 °C) of the chiral LC separation of norgestrel was studied using native CDs added to a methanol–water 1:1 eluent. α- or β-CDs gave no enantiodiscrimination, while baseline LC separation was achieved with γ-CD. ¹H NMR titrations of the racemate with α-CD showed only weak complexation, while β- or γ-CDs caused enantiomeric splitting of the steroid signals. Both the chiral selectivity values and their insignificant temperature dependence measured in the γ-CD-modified LC system were successfully reproduced by NMR titrations. This agreement corroborates the intuitive view for a wider temperature range that the stability difference of the diastereomeric γ-CD/steroid complexes governs this LC enantioseparation.     

LC Enantioseparation of Tryptophan Analogs on α-Cyclodextrin Stationary Phase

For the direct separation of enantiomers of tryptophan analogs, a native α-cyclodextrin-based Cyclobond III chiral stationary phase was examined. The separation conditions were optimized by variation of the different chromatographic parameters, and data are reported as retention factors, separation factors and resolutions. The nature and concentration of the buffers as mobile phase additivies exerted only slight effects on the retention and resolution. In all cases, the erythro isomers could be separated significantly better than the threo ones. With increasing hydrophobicity of the analytes, the retention time increased, but no clear trend could be observed between the resolution and the hydrophobicity. The sequence of elution of the enantiomers was in most cases determined by spiking the racemic samples with enantiorich analytes.     

Enantioseparation of 1-Phenyl-1,2,3,4-tetrahydroisoquinoline on Polysaccharide-Based Chiral Stationary Phases

The enantiomers of 1-phenyl-1,2,3,4-tetrahydroisoquinoline have been directly separated on polysaccharide-based chiral stationary phases (CSPs). The normal phase separation of (S)- and (R)-1-phenyl-1,2,3,4-tetrahydroisoquinoline was accomplished by screening of the immobilized Chiralpak IC column with different eluents. The effect of mobile phase type on retention, selectivity and resolution was studied. 2-Propanol or ethanol/n-hexane/ethanolamine mixtures were applied as mobile phases by screening of following polysaccharide-based immobilized (Chiralpak IA, Chiralpak IC) and coated (Lux Cellulose-1, Lux Cellulose-2, Lux Amylose-2) CSPs. Polar organic and reversed-phase conditions were also tested for direct enantioseparation of 1-phenyl-1,2,3,4-tetrahydroisoquinoline.     

Enantioselective Extraction System Containing Binary Chiral Selectors and Chromatographic Enantioseparation Method for Determination of the Absolute Configuration of Enantiomers of Cyclopentolate

The distribution coefficients and enantioseparation of cyclopentolate were studied in an extraction system containing d-tartaric acid ditertbutyl ester in organic phase and 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) in aqueous phase. Various parameters involved in the enantioseparation such as the type and the concentration of chiral selectors, pH value and a wide range of organic solvents were investigated. The maximum enantioselectivity (α = 2.13) and optimum distribution coefficients (K _R = 0.85, K _S = 0.40) were obtained under the following conditions: 0.10 mol/L HP-β-CD in aqueous phase and 0.20 mol/L d-tartaric acid ditertbutyl ester in decanol as organic phase. Cyclopentolate is present as a racemic mixture to the aqueous phase. The potentially different biological activities of cyclopentolate enantiomers have not been examined yet. Two chiral liquid chromatography methods have been developed for the direct separation of the enantiomers of cyclopentolate. First method was used for the quantification analysis of cyclopentolate enantiomers in aqueous phase. Second method used two chiroptical detectors: electronic circular dichroism (ECD) and optical rotation (OR) for the identification of individual cyclopentolate enantiomers from the organic phase enriched with (R)-enantiomer. The absolute stereochemistry was determined by means of the comparison of the experimental and computed ECD spectra and signs of OR. The ECD spectra of chiral analytes were measured on-line using HPLC-ECD technique.     

Phase Ratio and Equilibrium Constant in RP-HPLC Obtained from Octanol/Water Partition Constant Through Solvophobic Theory

The experimentally known dependence in RP-HPLC of the retention factor k′ on octanol/water partition coefficient (K _ow) has been examined based on solvophobic theory. The result showed that the dependence provides a means for the evaluation of phase ratio (Φ) of RP-HPLC columns, and of the equilibrium constant for a given compound and mobile phase. Using this theory, the phase ratio was evaluated for a set of seven different C18 columns (five having fully porous particles and two core–shell particles), and the equilibrium constants were calculated for four homologous series of compounds in two mobile phase systems. One mobile phase was methanol/aqueous solution of 0.1% H₃PO₄, and the other was acetonitrile/aqueous solution of 0.1% H₃PO₄. Besides providing the values for Φ for the evaluated columns, the results of the study indicated that for a specific composition of the mobile phase and for a given compound displaying only hydrophobic interactions, the equilibrium constant K(X) for different C-18 columns is basically the same. The data were further used to provide guidance in the selection of a chromatographic column for a specific separation based on K _ow values and chemical structure of the analytes. The study indicated that the separation of compounds with identical polar groups (or no polar groups) and with very close values for the K _ow cannot be achieved based only on hydrophobic interactions that dominate the separation on RP-type columns. Only column that displays polar interactions may provide a solution to such separations. For hydrocarbons with close K _ow values, the separation cannot be achieved even on columns with some polarity. On the other hand, even compounds with equal K _ow values, but with different functionalities can be separated on RP-HPLC columns without involving polar interactions. The compounds with different K _ow values are expected to be easily separated on RP-HPLC columns.     

Determination of the retention characteristics in thin-layer chromatography taking into account the actual position of the mobile phase front

To reveal the actual position of the mobile phase front, corresponding to the free volume of the layer in thin-layer chromatography, it is suggested to use a nonsorbable low-molecular-weight marker. The expressions for the capacity coefficient k′ equilibrium coefficient K _d , and corrected R _f values (R′_f) were obtained taking into account the correction of the position of the mobile phase front.     

An electrochemical chiral sensor based on amino-functionalized graphene quantum dots/β-cyclodextrin modified glassy carbon electrode for enantioselective detection of tryptophan isomers

An electrochemical chiral sensing platform based on amino-functionalized graphene quantum dots/β-cyclodextrin modified glassy carbon electrode (NH₂-GQDs/β-CD/GCE) was developed for enantioselective detection of tryptophan (Trp) isomers. NH₂-GQDs/β-CD/GCE showed high electrocatalytic activity and good analytical behavior toward the oxidation of Trp isomers. The oxidation peak potentials and oxidation peak currents of Trp isomers at NH₂-GQDs/β-CD/GCE surface were observed by differential pulse voltammetry. NH₂-GQDs/β-CD nanocomposite exhibited different binding ability for two Trp isomers and selectively bonded with d-Trp, resulting in the higher oxidation peak current of d-Trp at NH₂-GQDs/β-CD/GCE surface. Trp isomers exhibited different oxidation peak potentials at NH₂-GQDs/β-CD/GCE surface, and the peak potential separation between l-Trp and d-Trp was around 0.022 V, which was used for the enantioselective detection of Trp isomers. Under the optimum experimental conditions, the oxidation peak currents were linearly dependent on the concentrations of Trp isomers. The linear ranges of l-Trp and d-Trp were all from 1.0 to 30.0 μM with correlation coefficients of 0.9886 and 0.9800, respectively. The detection limits of l-Trp and d-Trp were 0.65 and 0.12 μM (3σ/K), respectively. Such NH₂-GQDs/β-CD/GCE displayed high anti-interference against some physiological substances, good reproducibility and excellent long-term stability toward Trp isomers detection in biomedical applications.     

HPLC Enantioseparation of Novel Spirobrassinin Analogs on the Cyclofructan Chiral Stationary Phases

A novel series of nine chiral analogs of spirobrassinin, which have potential biological activity, was separated for the first time on three different derivatized cyclofructan chiral stationary phases in the normal phase mode. The effects of mobile phase composition, the type and concentration of polar modifier, additives, and the analyte structure on the retention and enantioseparation were studied. The results proved that for cyclofructan-based chiral stationary phases, the R-naphthylethyl carbamate cyclofructan 6 provides the best separation efficiency for the analyzed compounds. The effect of temperature on the separation was also investigated and the corresponding thermodynamic parameters were evaluated from linear Van’t Hoff plots (lnk or lnα versus 1/T). It was found that the enantioseparation was enthalpy controlled. In addition, the elution order of the enantiomers was determined in all the cases.     

Potential functions of internal rotation in <Emphasis Type="Italic">n</Emphasis>-alkanes and its contribution to thermodynamic properties

The potential functions of braked internal rotation V(?) in n-alkanes (ethane, propane, butane, n-pentane, n-hexane, n-heptane) were calculated by ab initio and DFT methods with the 6-311++G(3df,3pd) basis set. The functions were approximated as a series of six cosines. The dependences of V(?) on the length of the hydrocarbon chain in n-alkanes were analyzed. The heights of the trans-cis and trans-gauche barriers and the differences between the energies of the trans and gauche conformers were calculated and compared with the experimental data. From the calculated geometric parameters and V(?), the contributions of the braked internal rotation to the enthalpy, entropy, heat capacity, and Gibbs free energy at 298 K were determined. The contributions of internal rotations are transferable within the framework of additive approaches. The generalized function V _av(?) for n-alkanes and averaged contributions of internal rotation of the C-C bonds and CH₃- and -CH₂- tops to the thermodynamic properties were suggested.     

Separation of β-blocker and amino acid enantiomers on a mixed chiral sorbent modified with macrocyclic antibiotics eremomycin and vancomycin

A mixed chiral sorbent based on silica with immobilized macrocyclic antibiotics eremomycin and vancomycin was synthesized. A possibility of the separation of enantiomers of β-blockers (metoprolol, pindolol, alprenolol, oxprenolol, labetalol, and atenolol) and amino acids (tryptophan, phenylalanine, DOPA, methionine, and acetyl glutamic acid) on this chiral sorbent by HPLC was studied. The influence of the composition of the mobile phase (pH of buffer solution, its concentration, content of organic modifier, and its nature) on the retention times of β-blocker and amino acid enantiomers, selectivity, and resolution of peaks was studied. It was shown that the mixed chiral sorbent has enantioselectivity to both classes of compounds, while silica modified with vancomycin has no ability to the separation of enantiomers of non-derivatized amino acids, and silica modified with eremomycin has no ability to the separation of β-blocker enantiomers. High values of resolution for amino acids (max Rs > 4) and β-blockers (max Rs > 1) were obtained.     

Kinetic study on extractive resolution of tropic acid by hydroxyethyl-β-cyclodextrin

Reactive extraction is proposed as a promising approach to the resolution of tropic acid (TA), where hydroxyethyl-β-cyclodextrin (HE-β-CD) is used as a selective extractant. The reaction between tropic acid and extractant occurred in the aqueous phase, and kinetic studies of reactive extraction have been investigated in a Lewis-type stirred cell. The batch experiments have been performed to study the process parameters affecting the extraction rate including agitation speed, interfacial area, pH value of aqueous phase, initial concentration of tropic acid enantiomers and initial concentration of extractant on the initial extraction rate. The kinetic equation of the forward rate was deduced at the experiment condition: r _TA+ = (4.041 × 10^?3)[TA₊][CD]²; r _TA? = (3.1234 × 10^?3)[TA_?][CD]².     

Preparation,characterization of new Co(II) and Cu(II) phthalocyanines and their catalytic performances in aerobic oxidation of substituted phenols

The host–guest inclusion complex of meta-cresol (m-cresol, liquid at room temperature, guest molecule) is synthesized with β-cyclodextrin (β-CD, host molecule). The inclusion complex is characterized by various techniques like nuclear magnetic resonance, powder X-ray diffraction, field emission scanning electron microscopy and spectroscopic (steady state as well as time-resolved) techniques. To the best of our knowledge, no literature data is available on the photophysical properties (especially fluorescence upon photoexcitation) of m-cresol in liquid media till date. Our spectroscopic studies exhibit some interesting photophysical properties of m-cresol and its inclusion complex with β-CD in different liquid media. The present work is important in view of the various potential applications of m-cresol in science, technology and medicine.     

Enantioselective Extraction System Containing Binary Chiral Selectors and Chromatographic Enantioseparation Method for Determination of the Absolute Configuration of Enantiomers of Cyclopentolate

The distribution coefficients and enantioseparation of cyclopentolate were studied in an extraction system containing d-tartaric acid ditertbutyl ester in organic phase and 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) in aqueous phase. Various parameters involved in the enantioseparation such as the type and the concentration of chiral selectors, pH value and a wide range of organic solvents were investigated. The maximum enantioselectivity (α = 2.13) and optimum distribution coefficients (K _R = 0.85, K _S = 0.40) were obtained under the following conditions: 0.10 mol/L HP-β-CD in aqueous phase and 0.20 mol/L d-tartaric acid ditertbutyl ester in decanol as organic phase. Cyclopentolate is present as a racemic mixture to the aqueous phase. The potentially different biological activities of cyclopentolate enantiomers have not been examined yet. Two chiral liquid chromatography methods have been developed for the direct separation of the enantiomers of cyclopentolate. First method was used for the quantification analysis of cyclopentolate enantiomers in aqueous phase. Second method used two chiroptical detectors: electronic circular dichroism (ECD) and optical rotation (OR) for the identification of individual cyclopentolate enantiomers from the organic phase enriched with (R)-enantiomer. The absolute stereochemistry was determined by means of the comparison of the experimental and computed ECD spectra and signs of OR. The ECD spectra of chiral analytes were measured on-line using HPLC-ECD technique.

     

High-Performance Liquid Chromatographic Separation of Stereoisomers of <Emphasis Type="Bold">β</Emphasis>-Amino Acids and a Comparison of Separation Efficiencies on Chirobiotic T and TAG Columns

Direct and indirect reversed-phase high-performance liquid chromatographic methods were developed for the separation of enantiomers of seventeen unnatural β-amino acids, including several β-3-homo amino acids. The direct separations of the underivatized analytes were performed on chiral stationary phases containing macrocyclic glycopeptide antibiotic teicoplanin (Chirobiotic T column) and teicoplanin aglycone (Chirobiotic TAG column). The indirect method involved pre-column derivatization with two new chiral derivatizing agents, (1S,2S)-1,3-diacetoxy-1-(4-nitrophenyl)-2-propylisothiocyanate, (S,S)-DANI and (S)-N-(4-nitrophenoxycarbonyl)phenylalanine methoxyethyl ester, (S)-NIFE. The different methods were compared in systematic chromatographic examinations. The effects of organic modifier, mobile phase composition, pH and flow rate on the separation were investigated.