Combination of capillary electrophoresis and molecular modeling to study the enantiomer affinity pattern between β‐blockers and anionic cyclodextrin derivatives in a methanolic and water background electrolyte

In order to have deep insights into the mechanisms of enantiomer affinity pattern in both aqueous and non‐aqueous systems, an approach combining capillary electrophoresis and molecular modeling was undertaken. A chiral β‐blocker; acebutolol, was enantioseparated in aqueous capillary electrophoresis and non‐aqueous capillary electrophoresis using two anionic β‐cyclodextrin derivatives. The enantiomer affinity pattern of acebutolol was found to be opposite when an aqueous background electrolyte was replaced with non‐aqueous background electrolyte in the presence of heptakis(2,3‐di‐O‐acetyl‐6‐sulfo)‐β‐cyclodextrin but remained the same in the presence of heptakis(2,3‐di‐O‐methyl‐6‐sulfo)‐β‐cyclodextrin. Molecular docking of acebutolol into two β‐cyclodextrin derivatives indicated two distinct binding modes called ‘up’ and ‘down’ conformations. After structure optimization by molecular dynamics and energy minimization, both enantiomers of acebutolol were preferred to the ‘up’ conformation with heptakis(2,3‐di‐O‐methyl‐6‐sulfo)‐β‐cyclodextrin while ‘down’ conformation with heptakis(2,3‐di‐O‐acetyl‐6‐sulfo)‐β‐cyclodextrin. The further calculation of the complex energy with solvent effect indicated that heptakis(2,3‐di‐O‐acetyl‐6‐sulfo)‐β‐cyclodextrin had higher affinity to S‐acebutolol than R‐acebutolol in non‐aqueous capillary electrophoresis while it showed better binding to R‐acebutolol in aqueous capillary electrophoresis. However, the heptakis(2,3‐di‐O‐methyl‐6‐sulfo)‐β‐cyclodextrin bound better to R‐acebutolol in both aqueous and non‐aqueous capillary electrophoresis, implying that the binding mode played more important role in chiral separation of heptakis(2,3‐di‐O‐methyl‐6‐sulfo)‐β‐cyclodextrin while the solvent effect had prevailing impact on heptakis(2,3‐di‐O‐acetyl‐6‐sulfo)‐β‐cyclodextrin.     

卡替诺尔和氟西汀对映体的高效毛细管电泳分离

考察了以羧甲基－β－（环糊精－β－CD）、β－环糊清（β－CD）、羟丙基－β－环糊精（HP－β－CD）、二甲基－β－环糊精（DM－β－CD)为手性选择剂,在50mmol/L醋酸三乙胺缓冲溶液中分离卡替诺尔和氟西汀对映体。该文还通过考察手性选择剂的浓度、背景电解质的酸度、背景电解质的类型等因素对映体手性分离的影响,对分离条件进行了优化,初步探讨了手性识别机理。实验结果表明：用约4mmol/L的CM－β－CD分离氟西汀和卡替诺尔对映体,能使对映体达到良好分离,不仅节约了分析成本,也简化了分析过程。     

Development of a capillary electrophoresis method for the determination of the chiral purity of dextromethorphan by a dual selector system using quality by design methodology

Dextromethorphan is a centrally acting antitussive drug, while its enantiomer levomethorphan is an illicit drug with opioid analgesic effects. As capillary electrophoresis has been proven as an ideal technique for enantiomer analysis, the present study was conducted in order to develop a capillary electrophoresis‐based limit test for levomethorphan. The analytical target profile was defined as a method that should be able to determine levomethorphan with acceptable precision and accuracy at the 0.1 % level. From initial scouting experiments, a dual selector system consisting of sulfated β‐cyclodextrin and methyl‐α‐cyclodextrin was identified. The critical process parameters were evaluated in a fractional factorial resolution IV design followed by a central composite face‐centered design and Monte Carlo simulations for defining the design space of the method. The selected working conditions consisted of a 30/40.2 cm, 50 μm id fused‐silica capillary, 30 mM sodium phosphate buffer, pH 6.5, 16 mg/mL sulfated β‐cyclodextrin, and 14 mg/mL methyl‐α‐cyclodextrin at 20°C and 20 kV. The method was validated according to ICH guideline Q2(R1) and applied to the analysis of a capsule formulation. Furthermore, the apparent binding constants between the enantiomers and the cyclodextrins as well as complex mobilities were determined to understand the migration behavior of the analytes.     

Chiral separation of benzothiazole derivatives of amino acids using capillary zone electrophoresis

A method for the separation of enantiomers of leucine and phenylalanine benzothiazole derivatives as potential antimicrobial agents was developed using capillary zone electrophoresis with a dual cyclodextrin (CD) system. The best resolution of enantiomers was achieved in 100 mmol/L phosphate background electrolyte (pH 3.5) with the dual CD system consisting of 10 mmol/L of β‐CD with 10 mmol/L of 2‐hydroxypropyl‐β‐cyclodextrin for leucine derivative and 10 mmol/L of 2‐hydroxypropyl‐γ‐cyclodextrin for phenylalanine derivative, respectively. Under the optimal conditions, the highest enantioresolution of 1.25 was achieved in a noncoated‐fused silica capillary at 17°C and 24 kV applied voltage.     

Enantioseparation of binaphthol and its mono derivatives by cyclodextrin-modified capillary zone electrophoresis

Chiral binaphthols belong to the group of most effective ligands for asymmetrical catalysis. In this context, various binaphthols presenting original substituents have been synthesized. Their study through capillary electrophoresis is the object of this work. The literature dedicated to the separation of atropisomers by capillary electrophoresis, corresponding only to binaphthol, reveals that its enantioseparation is always delicate because of the influence of many factors and the resolutions obtained are weak. Therefore, for a structured optimization, we first successfully evaluated the acidity constants of different binaphthols by means of capillary electrophoresis. With these known physicochemical characteristics, we could successfully carry out enantiomeric separations of the different binaphthols at pH 11.5, practically in completely ionized form, in phosphate medium, and in the presence of cyclodextrin (CD), with analysis times lower than 8min. The nature of CDs (alpha-CD, beta-CD, gamma-CD, hydroxypropyl-alpha-cyclodextrin (HP-alpha-CD), HP-beta-CD, HP-gamma-CD and trimethyl-beta-CD (TM-beta-CD)) and other factors in relation to enantiomeric resolution (applied voltage, nature and concentration of the electrolyte, and concentration of cyclodextrin) were optimized. These studies allowed us to determine the optimal conditions of separation (concentration and nature of CD) for each of the studied binaphthols. It is necessary to mention that, for the 1,1'-binaphthyl-2,2'-diol (Binol) at pH 11.5, the S atropisomer always migrated first, regardless of the nature and concentration of the cyclodextrin used. Moreover, an inversion in elution order of the two atropisomers as a function of pH was observed with gamma-CD (pH range: 10-11.5). The R atropisomer migrated first at pH 10. At pH 10.8 the migration order of the two atropisomers of Binol was reversed as a function of gamma-CD concentration. Finally, the addition of chiral ionic liquids (R(-)-1-hydroxy-N,N,N-trimethylbutan-2-aminium bis(trifluoromethylsulfonyl)imide and S(+)-tetrabutylammonium camphorsulfonate) was conducted. In the case of S(+)-tetrabutylammonium camphorsulfonate, a weak antagonistic effect was observed with modeling the evolution of enantiomeric resolution by means of the experimental design, while in the case of R(-)-1-hydroxy-N,N,N-trimethylbutan-2-aminium bis(trifluoromethylsulfonyl)imide the effect was neutral.     

High Resolution of Racemic Mandelic Acid through a Method of Bubble Fractionation

A method of bubble fractionation was developed for the resolution of racemic mandelic acid (MA), using 2‐hydroxypropyl‐β‐cyclodextrin (HP‐β‐CD) as chiral collector. The influences of concentration of HP‐β‐CD, reflux equilibriation time, gas flow rate, packing height of column and pH of buffer on resolution performance were investigated, respectively. According to the similar physical behavior of bubble fractionation and chemical reaction processes, the equivalent chemical reaction constant was introduced. The resolution process was preliminarily analyzed by means of kinetics. The results show that the enantiomeric excess of 60.7% can be obtained under the optimal conditions. The process could be regarded as a first order chemical reaction, where the equivalent speed constant was k_l=0.00376. This method is helpful for realizing high resolution and linear amplification of device.     

Affinity capillary electrophoresis method for investigation of bile salts complexation with sulfobutyl ether‐β‐cyclodextrin

Sulfobutyl ether‐β‐cyclodextrin (SBEβCD) is utilized in preformulation and drug formulation as an excipient for solubilization of drugs with poor aqueous solubility. Approximately seven negative charges of SBEβCD play a role with respect to solubilization and complexation, but also have an influence on the ionic strength of the background electrolyte when the cyclodextrin is used in capillary electrophoresis. Mobility‐shift affinity capillary methods for investigation of the complexation of taurocholate and taurochenodeoxycholate with the negatively charged cyclodextrin derivative applying constant power and ionic strength conditions as well as constant voltage and varying ionic strength were investigated. A new approach for the correction of background electrolyte ionic strength was developed. Mobility‐shift affinity capillary electrophoresis experiments obtained at constant voltage and constant power settings were compared and found to provide binding parameters that were in good agreement upon correction. The complexation of taurochenodeoxycholate with SBEβCD was significantly stronger than the corresponding interaction involving taurocholate. The obtained stability constants for the bile salts were in the same range as those previously reported for the interaction with neutral β‐cyclodextrins derivatives, i.e. the positions of the negative charges on SBEβCD and the bile salts within the complex did not lead to significant electrostatic repulsion.     

Cyclodextrins in polymer synthesis: polymerization of methyl methacrylate under atom‐transfer conditions (ATRP) in aqueous solution

Host guest complexes of methyl methacrylate (MMA) and randomly methylated β‐cyclodextrin (m‐β‐CD, 1 a ) were polymerized in aqueous medium using atom‐transfer radical polymerization. Ethyl 2‐bromoisobutyrate (EBIB) was used as an initiator, copper(I) bromide as the catalyst, and bipyridine (bipy) or 4,4′‐di‐(5‐nonyl)‐2,2´‐bipyridine (dNbipy) as ligands. The unthreading of m‐β‐CD during the polymerization led to water‐insoluble poly(methyl methacrylate) (PMMA). It was found that using dNbipy resulted in higher monomer conversion than using bipy as the ligand under similar conditions. Furthermore, it is shown that the polymerization of MMA under these conditions has a living character. The polymers obtained have a much lower polydispersity than those obtained from conventional free‐radical polymerization. Also, the block copolymerization of PMMA bearing a bromoester end group with CD‐complexed styrene ( 2 a ) was carried out under ATRP conditions in aqueous medium.     

Chiral separation of deprenyl-N-oxide isomers by capillary electrophoresis using various cyclodextrin derivatives

Chiral separation of deprenyl-N-oxide isomers is presented using capillary electrophoresis in the presence of various cyclodextrin (CD) derivatives. This recently identified metabolite of R-(-)-deprenyl may possess desirable pharmacological activities. The effect of the cavity size and the substituents of the CD are examined on the enantiomer resolution of the compound having an asymmetric center on a heteroatom. The importance of hydrophilic or hydrogen bonding interaction, as well as the position of the interacting groups is demonstrated. Outstanding selectivity and resolution values are achieved using the chargeable carboxymethyl-beta-CD. 2-Hydroxypropyl-beta-CD is also suitable for the enantiomer separation of the analyte. Native beta-CD and carboxyethyl-beta-CD provide only poor enantioselectivity, whereas heptakis-(2,6-di-O-methyl)-beta-CD is capable of separating only the diastereomers. No chiral resolution can be observed in the presence of gamma-CD.     

Rapid development of the enantiomeric separation of beta-blockers by capillary electrophoresis using an experimental design approach.

A rapid method for determining the separation conditions for chiral resolution of eleven beta-blocking drug substances by capillary electrophoresis is described, using an experimental design approach. An acidic phosphate-triethanolamine buffer and an uncoated fused-silica capillary were used for all experiments. Several modified cyclodextrins were applied as chiral selectors: sulfobutyl ether beta-cyclodextrin (SBE-beta CD), dimethyl beta-cyclodextrin (DM-beta CD), carboxymethyl beta-cyclodextrin (CM-beta CD), and hydroxypropyl beta-cyclodextrin (HP-beta CD). Two different fractional factorial experimental designs were applied: (1) a design examining four factors at three levels (3(4-2)) and (2) one examining three factors at two levels (2(3-1)). The factors studied were: type of cyclodextrin, cyclodextrin concentration, pH of the background electrolyte and percentage of organic modifier. Enough resolution for the separation of the enantiomers and even for their quantification was reached. The same scheme is proposed when a fast chiral separation method needs to be developed for other drug families.     

Study of binary systems of β-cyclodextrin with a highly potential anti-mycobacterial drug

The present research paper is dedicated to the obtaining and physicochemical characterization of a highly potential anti-mycobacterial drug candidate with β-cyclodextrin (βCD). The active substance is a 1,3,4-oxadiazole derivative, 2-phenyl-5-{[(2-phenyl-1,3-dioxolan-2-yl)methyl]sulfanyl}-1,3,4-oxadiazole, further named DIOX. DIOX–βCD binary systems were obtained as a physical mixture and a lyophilized product with molar ratio between the main components equal to 1:1 and 1:2. The obtained systems were submitted to physicochemical characterization applying the following instrumental methods: infrared spectrometry, differential scanning calorimetry, and X-ray crystallographic analysis. Besides, a molecular modeling analysis has been performed. The research data suggested certain intermolecular interaction between DIOX and βCD, suggesting formation of a three-molecular inclusion complex DIOX:βCD, including one DIOX molecule and two molecules of βCD. The main parts of the DIOX molecule included in the hydrophobic cavity of the cyclodextrin molecules most probably are dioxolane cycle and two benzene rings.     

Chiral capillary electrophoresis with cationic pyrrolidinium‐β‐cyclodextrin derivatives as chiral selectors

New single‐isomer, cationic β‐cyclodextrins, including mono‐6‐deoxy‐6‐pyrrolidine‐β‐cyclodextrin chloride (pyCDCl), mono‐6‐deoxy‐6‐(N‐methyl‐pyrrolidine)‐β‐cyclodextrin chloride (N‐CH₃‐pyCDCl), mono‐6‐deoxy‐6‐(N‐(2‐hydroxyethyl)‐pyrrolidine)‐β‐cyclodextrin chloride (N‐EtOH‐pyCDCl), mono‐6‐deoxy‐6‐(2‐hydroxymethyl‐pyrrolidine)‐β‐cyclodextrin chloride (2‐MeOH‐pyCDCl) were synthesized and used as chiral selectors in capillary electrophoresis for the enantioseparation of carboxylic and hydroxycarboxylic acids and dansyl amino acids. The unsubstituted pyCDCl exhibited the greatest resolving ability. Most analytes were resolved over a wide range of pH from 6.0 to 9.0 with this chiral selector. In general, increasing pH led to a decrease in resolution. The effective mobilities of all the analytes were found to decrease with increasing CD concentration. The optimal concentration for most carboxylic acids and dansyl amino acid was in the range 5–7.5 mM and >15 mM for hydroxycarboxylic acids. ¹H NMR experiments provided direct evidence of inclusion in the CD cavity.     

Preparation and evaluation of monodispersed,submicron, non‐porous silica particles functionalized with β‐CD derivatives for chiral‐pressurized capillary electrochromatography

Submicron, non‐porous, chiral silica stationary phase has been prepared by the immobilization of functionalized β‐CD derivatives to isocyanate‐modified silica via chemical reaction and applied to the pressurized capillary electrochromatography (pCEC) enantio‐separation of various chiral compounds. The submicron, non‐porous, cyclodextrin‐based chiral stationary phases (sub_μm‐CSP2) exhibited excellent chiral recognition of a wide range of analytes including clenbuterol hydrochloride, mexiletine hydrochloride, chlorpheniramine maleate, esmolol hydrochloride, and metoprolol tartrate. The synthesized submicron particles were regularly spherical and uniformly non‐porous with an average diameter of around 800 nm and a mean pore size of less than 2 nm. The synthesized chiral stationary phase was packed into 10 cm × 100 μm id capillary columns. The sub_μm‐CSP2 column used in the pCEC system showed better separation of the racemates and at a higher rate compared to those used in the capillary liquid chromatography mode (cLC) system. The sub_μm‐CSP2 possessed high mechanical strength, high stereoselectivity, and long lifespan, demonstrating rapid enantio‐separation and good resolution of samples. The column provided an efficiency of up to 170 000 plates/m for n‐propylbenzene.     

Determination of quinocide as impurity in primaquine tablets by capillary zone electrophoresis

A capillary zone electrophoretic method has been developed and validated for the determination of the impurity quinocide (QC) in the antimalarial drug primaquine (PQ). Different buffer additives such as native cyclodextrins and crown ethers were evaluated. Promising results were obtained when either β‐cyclodextrin (β‐CD) or 18‐crown‐6 ether (18C6) were used. Their separation conditions such as type of buffer and its pH, buffer additive concentration, applied voltage capillary temperature and injection time were optimized. The use of 18C6 offers slight advantages over β‐CD such as faster elution times and improved resolution. Nevertheless, migration times of less than 5 min and resolution factors (R_s) in the range of 2–4 were obtained when both additives were used. The method was validated with respect to selectivity, linearity, limits of detection and quantitation, analytical precision (intra‐ and inter‐day variability) and repeatability. Concentrations of 2.12 and 2.71% (w/w) of QC were found in pharmaceutical preparations of PQ from two different manufacturers. A possible mechanism for the successful separation of the isomers is also discussed. Copyright © 2008 John Wiley & Sons, Ltd.     

环糊精与两种有机物的包合物的不同光解特性

The effectiveness of the solubilization and photodegradation of β-cyclodextrin （β-CD） and hydroxypropyl-β- cyclodextrin （HPCD） on two hydrophobic organic compounds （HOC） of methyl parathion and pentachlorophenol was investigated. The results indicate that the solubilization or photodegradation of two HOC were influenced by complexing with β-CD or HPCD. The solubility of pentachlorophenol （PCP） was increased linearly with β-CD concentration. The solubility of methyl parathion （MPT） was increased with the increase of β-CD concentration initially, however, as the β-CD concentration was enhanced above 3 g/L, the solubility was decreased with increase of β-CD concentration. The solubilities of two HOC were increased linearly with the increase of HPCD concentration. Although the photodegradation of MPT was improved, the photodegradation of PCP was restrained by complexation of HOC with β-CD or HPCD. In a word, the effectiveness of photodegradation or solubilization of HPCD was more significant than that of β-CD. One potential application of such a method was the in situ remediation of hydrophobic organic pollutants in contaminated soil and groundwater or industrial waste streams.     

Cyclodextrins in polymer synthesis: free radical polymerization of cyclodextrin host‐guest complexes of methyl methacrylate or styrene from homogenous aqueous solution

The polymerization of methylated β‐cyclodextrin (m‐β‐CD) 1 : 1 host‐guest compounds of methyl methacrylate (MMA) ( 1 ) or styrene ( 2 ) is described. The polymerization of complexes 1 a and 2 a was carried out in water with potassium peroxodisulfate (K₂S₂O₈)/sodium hydrogensulfite (NaHSO₃) as radical redox initiator at 60°C. Unthreading of m‐β‐CD during the polymerization led to water‐insoluble poly(methyl methacrylate) (PMMA) ( 3 ) and polystyrene ( 4 ). By comparison, analogously prepared polymers from uncomplexed monomers 1 and 2 in ethanol as organic solvent with 2,2′‐azoisobutyronitrile (AIBN) as radical initiator showed significantly lower molecular weights and were obtained in lower yields in all cases. Polymerization of m‐β‐CD complexed MMA in water, initiated with 2,2′‐azobis(N,N ′‐dimethyleneisobutyroamidine) dihydrochloride, occurred much faster than the polymerization of uncomplexed MMA in methanol under similar conditions. Furthermore, it was shown, that the precipitation polymerization of complexed MMA from homogeneous aqueous solution can be described by equations (P_n^–1 ∝ lsqb;Irsqb;^0.5) similar to those for classical polymerization in solution.     

甲基绿的电化学行为及其与环糊精形成超分子体系研究

应用“电流法”研究了甲基绿—环糊精超分子体系 ,测定了甲基绿和 4种环糊精的包结常数 .其包结能力为HP_β_CD >β_CD >SBE_β_CD >CM_β_CD ,包结比为 1 :2 (甲基绿 :环糊精 )但甲基绿与α_CD和γ_CD不能形成超分子体系 .此外 ,本文还探讨了甲基绿的电极反应机理 ,初步表明 ,甲基绿在电极上还原的电子转移数为 4 ,有 2个质子参与反应     

2，6-二-O-乙氧乙基-3-O-三氟乙酰基-β-环糊精气相色谱手性固定相的制备及应用

将β-环糊精的2,6-位引入乙氧乙基,3-位引入三氟乙酰基,合成了新的环糊精衍生物2,6-二-O-乙氧乙基-3-O-三氟乙酰基-β-环糊精,并采用静态法涂渍毛细管气相色谱柱,考察了毛细管柱的柱性能和分离性能。结果表明,该固定相对G rob试剂、苯的二取代位置异构体氯甲苯、硝基甲苯和溴甲苯以及10种手性化合物如α-取代丙酸酯化合物、1-(2′-硝基苯基)-乙醇、α-甲基-对氯苯乙腈和丙炔醇酮乙酸酯等具有良好的分离效果。其中,对α-甲磺酰基丙酸酯对映体的拆分效果最好;对α-取代丙酸的甲酯衍生物的分离效果优于乙酯衍生物;对α-羟基取代丙酸酯的分离效果优于α-卤代丙酸酯。     

Cyclodextrin‐mediated capillary electrophoresis enantioseparation of dansylated β‐amino acids with bicyclo[2.2.2]octane,bicyclo[3.1.1]heptane and cyclopenta[d][1,2]oxazole core structures

The present study investigated the separation of bicyclic β‐amino acids with bicyclo[2.2.2]octane, bicyclo[3.1.1]heptane and cyclopenta[d][1,2]oxazole core structures by capillary electrophoresis using native cyclodextrins as well as neutral and charged derivatives as chiral selectors. The amino acids were derivatized with dansyl chloride to provide a UV chromophore. Separations were carried out at 20°C in a 48.5/40 cm, 50 µm fused‐silica capillary at an applied voltage of 20 kV. Fifty millimolar sodium phosphate background electrolytes pH 2.5 and 7.2 containing either 5 or 30 mg/mL of the CDs were used. For the majority of the investigated CDs, enantioseparations could only be achieved at pH 2.5 when the analytes are positively charged. Successful enantioseparations as negatively charged analytes at pH 7.2 were only observed for few compounds. In the case of methyl‐γ‐cyclodextrin, opposite enantiomer migration order was observed in pH 2.5 or 7.2 background electrolytes. Dependence of the enantiomer migration order on the size of the cavity of the cyclodextrins was also found. Furthermore, the degree of methylation of β‐cyclodextrin derivatives affected the migration order of several analyte enantiomers.     

Cyclodextrin Derivatives in GC Separation of Racemic Mixtures of Volatiles. Part XIV: Some Applications of Thick-Film Wide-Bore Columns to Enantiomer GC Micropreparation

Representative examples of preparative GC isolation of pure enantiomers in amounts ranging from 0.5 to 2 mg, with thick-film wide-bore open tubular columns coated with cyclodextrin (CD) diluted in polysiloxanes, are described. Two 25 m×0.53 mm i.d. columns, coated with 3 μm of 30% 2,6-di-O-methyl-3-O-pentyl-β-CD/OV-1701 and 2 μm of 30% 2,3-di-O-acetyl-6-O-tert- butyldimethylsilyl-β-CD/PS-086 as stationary phases, were used. Methyl 3-hydroxyhexanoate, methyl 2-methylbutanoate, δ-hexalactone, δ-octalactone, and γ-decalactone (the latter from a nature-identical mango aroma), were submitted to automated micropreparative GC. Average yields of about 30% were achieved when amounts above 20 μg/μl were injected, and an enantiomeric purity above 90% was obtained.