High-performance liquid chromatographic enantioseparation of amino compounds on newly developed cyclofructan-based chiral stationary phases

Direct separation of the enantiomers of amino acid amines, amino alcohols, and diamines was performed on recently developed chiral stationary phases containing isopropyl carbamate-cyclofructan 6 (IP-CF6), (R)-naphthylethylcarbamate cyclofructan 6 (RN-CF6), or dimethylphenylcarbamate cyclofructan 7 (DMP-CF7) as chiral selectors, using n-hexane/alcohol/TFA as mobile phase. The effects of the mobile phase composition, the nature and concentrations of the alcoholic and acidic modifiers, and the structures of the analytes on the retention and resolution were investigated. In some cases, separations were carried out at constant mobile phase composition in the temperature range 5-40 °C. Thermodynamic parameters and T(iso) values were calculated from plots of lnk versus 1/T. It was found that the enantioseparations were enthalpy driven. The sequences of elution of the stereoisomers were determined but no general rule could be established.     

High-performance liquid chromatographic enantioseparation of aminonaphthol analogs on polysaccharide-based chiral stationary phases

High-performance liquid chromatographic methods were developed for the separation of the enantiomers of five new aminonaphthol analogs possessing two chiral centers. The direct separations were performed on chiral stationary phases containing either amylose-tris-3,5-dimethylphenyl carbamate (Kromasil^® AmyCoat™ column) or cellulose-tris-3,5-dimethylphenyl carbamate (Kromasil^® CelluCoat™ column) as chiral selector. The experimental data are utilized to discuss the effects of the mobile phase composition, the nature of the alcoholic modifier and the specific structural features of the analytes on the retention and separation. The elution sequence was determined in all cases; no general regularities could be established.     

Retention mechanism of high-performance liquid chromatographic enantioseparation on macrocyclic glycopeptide-based chiral stationary phases

The development of methods for the separation of enantiomers has attracted great interest in the past 20 years, since it became evident that the potential biological or pharmacological applications are mostly restricted to one of the enantiomers. In the past decade, macrocyclic antibiotics have proved to be an exceptionally useful class of chiral selectors for the separation of enantiomers of biological and pharmacological importance by means of high-performance liquid chromatography (HPLC), thin-layer chromatography and electrophoresis. The glycopeptides avoparcin, teicoplanin, ristocetin A and vancomycin have been extensively used as chiral selectors in the form of chiral bonded phases in HPLC, and HPLC stationary phases based on these glycopeptides have been commercialized. In fact, the macrocyclic glycopeptides are to some extent complementary to one another: where partial enantioresolution is obtained with one glycopeptide, there is a high probability that baseline or better separation can be obtained with another. This review sets out to characterize the physicochemical properties of these macrocyclic glycopeptide antibiotics and, through their application, endeavors to demonstrate the mechanism of separation on macrocyclic glycopeptides. The sequence of elution of the stereoisomers and the relation to the absolute configuration are also discussed.     

High-performance liquid chromatographic enantioseparation of drugs containing multiple chiral centers on polysaccharide-type chiral stationary phases

Enantioseparation of drugs with multiple chiral centers is challenging. This article describes resolution of some drugs with multiple chiral centers using polysaccharide-type chiral stationary phases. Also, the use of the column-switching technique is demonstrated to achieve the resolution of this type of compounds.     

High-performance liquid chromatographic enantioseparation of monoterpene-based 2-amino carboxylic acids on macrocyclic glycopeptide-based phases

The enantiomers of five monoterpene-based 2-amino carboxylic acids were directly separated on chiral stationary phases containing macrocyclic glycopeptide antibiotics such as teicoplanin (Astec Chirobiotic T and T2) and teicoplanin aglycone (Chirobiotic TAG) as chiral selectors. The effects of pH, the mobile phase composition, the structure of the analyte and temperature on the separations were investigated. Experiments were performed at constant mobile phase compositions in the temperature range 10–40 °C to study the effects of temperature and thermodynamic parameters on separations. Apparent thermodynamic parameters and T_iso values were calculated from plots of ln k or ln α versus 1/T. Some mechanistic aspects of the chiral recognition process are discussed with respect to the structures of the analytes. It was found that the enantioseparations were in most cases enthalpy driven. The sequence of elution of the enantiomers was determined in all cases.     

Non-aqueous capillary electrophoretic enantioseparation of N-derivatized amino acids using cinchona alkaloids and derivatives as chiral counter-ions

A non-aqueous capillary electrophoretic method developed with quinine and tert.-butyl carbamoylated quinine as chiral selectors for the enantioseparation of N-protected amino acids was applied to the investigation of other quinine derivatives as chiral additives. The optimum composition of the background electrolyte was found to be 12.5 mM ammonia, 100 mM octanoic acid and 10 mM chiral selector in an ethanol-methanol (60:40, v/v) mixture. Under these conditions, a series of chiral acids, as various benzoyl, 3,5-dinitrobenzoyl and 3,5-dinitrobenzyloxycarbonyl amino acid derivatives were investigated with regards to selectand-selector relationships and enantioselectivity employing quinine, quinidine, cinchonine, cinchonidine, tert.-butyl carbamoylated quinine, tert.-butyl carbamoylated quinidine, dinitrophenyl carbamoylated quinine and cyclohexyl carbamoylated quinine as chiral selector.     

Strategies for enantioseparations of catecholamines and structurally related compounds by capillary zone electrophoresis using sulfated beta-cyclodextrins as chiral selectors

Strategies for simultaneous enantioseparations of three catecholamines (DL-norepinephrine, DL-epinephrine, and DL-isoproterenol) and three structurally related compounds (DL-octopamine, DL-synephrine, and DL-norephedrine) by CZE using sulfated beta-CDs as chiral selectors were investigated. Four different separation modes were attempted: (I) using randomly sulfate-substituted beta-CD (MI-S-beta-CD) at relatively low concentrations in a high-concentration phosphate buffer at low pH in the normal polarity mode, (II) using MI-S-beta-CD at high concentrations at low pH in the reversed polarity mode, (III) using MI-S-beta-CD at moderately high concentrations in a phosphate buffer at neutral pH in the normal polarity mode, and (IV) using the single isomer heptakis(2,3-dihydroxy-6-O-sulfo)-beta-CD (SI-S-beta-CD) at low to moderately high concentrations in a high-concentration BGE at low pH in the normal polarity mode. Among them, enantioseparation of these cationic solutes was best achieved under the conditions of mode (II). In mode (II) and mode (III), temperature is an important factor affecting the enantioresolution of norepinephrine. In mode (I) and mode (IV), the use of a high-concentration BGE (150-200 mM) is crucial for effective enantioseparation of these cationic solutes with sulfated beta-CDs. Comparative studies of enantioseparations of these cationic solutes with MI-S-beta-CD and SI-S-beta-CD reveal that the sulfate substituents of MI-S-beta-CD located at the C(2)- position interact strongly with the diol moiety of catecholamines.     

High-performance liquid chromatographic enantioseparation of 2-aminomono- and dihydroxycyclopentanecarboxylic and 2-aminodihydroxycyclohexanecarboxylic acids on macrocyclic glycopeptide-based phases

The direct separation of the enantiomers of four 2-aminomono- or dihydroxycyclopentanecarboxylic acids and four 2-aminodihydroxycyclohexanecarboxylic acids was performed on chiral stationary phases containing macrocyclic glycopeptide antibiotics such as teicoplanin (Astec Chirobiotic T and T2), teicoplanin aglycone (Chirobiotic TAG) or ristocetin A (Chirobiotic R) as chiral selectors. The effects of the nature of organic modifiers, the pH, the mobile phase composition and the structures of the analytes on the separation were investigated. Chirobiotic TAG, and in some cases Chirobiotic T, proved to be the most useful of these columns. The elution sequence was determined in most cases.     

High-performance liquid chromatographic,capillary electrophoretic and capillary electrophoretic-electrospray ionisation mass spectrometric analysis of selected alkaloid groups

Systems for efficient separation of selected alkaloid groups by high performance liquid chromatography (HPLC), capillary electrophoresis (CE) and capillary electrophoresis coupled with electrospray ionisation mass spectrometry (CE-ESI-MS) are described. The optimized HPLC system was applied for the separation of 23 standard indole alkaloids as well as for qualitative and quantitative analyses of crude alkaloid extracts of Rauvolfia serpentina X Rhazya stricta hybrid cell cultures. The developed conditions for CE analysis proved to be efficient for separation of mixtures of standard indole and beta-carboline alkaloids. The described buffer system is also applicable in the combination of CE with electrospray ionisation mass spectrometry. This analytical technique allowed the separation and identification of components of standard indole alkaloid mixture as well as crude extracts of R. serpentina roots, R. serpentina cell suspension cultures and cortex of Aspidosperma quebracho-blanco. The influence of buffer composition and analyte structures on separation is discussed.     

New derivatives of cyclodextrins as chiral selectors for the capillary electrophoretic separation of dichlorprop enantiomers

-, β- and γ-cyclodextrins (CDs), as well as some of their chemical derivatives, have been tested as chiral resolving agents for the capillary zone electrophoretic resolution of the racemic herbicide dichlorprop, (±)-2-(2,4-dichlorophenoxy)propionic acid, of which only the (+)-isomer is herbicidally active. The complexation constants of the herbicide enantiomers with the cyclodextrin host molecules have been calculated from the electrophoretic migration time data at variable cyclodextrin concentration. The experimental results showed that several of the investigated CDs allowed dichlorprop enantiomer resolution. In particular, a newly synthesised ethylcarbonate derivative of β-CD showed the best enantiomer resolution properties among the tested compounds, while the remaining ones showed inferior or no performances at all. The calculated inclusion constants allowed identification of the best conditions for enantioresolution, and an explanation of the different complexation properties of the investigated compounds has been proposed on the basis of molecular modeling.     

Validated high-performance liquid chromatographic enantioseparation of selenomethionine using isothiocyanate based chiral derivatizing reagents

A high-performance liquid chromatographic (HPLC) method for enantioseparation of selenomethionine (SeMet) was developed using two isothiocyanate-based chiral derivatizing reagents [(R)-methyl benzyl isothiocyanate (MBIC) and (S)-1-(1-naphthyl) ethyl isothiocyanate (NEIC)] and UV detection. Diastereomers of selenomethionine were synthesized either via stirring (using MBIC) or by microwave irradiation (using NEIC). Derivatization conditions were optimized and the synthesized diastereomers were successfully resolved using triethyl ammonium phosphate buffer and acetonitrile on a reversed-phase column. The method was validated for accuracy, precision and limit of detection. The mechanism of separation is also discussed.     

高效液相色谱手性流动相添加法拆分阿卓乳酸对映体

采用C₁₈反相色谱柱,以磺丁基醚-β-环糊精（SBE-β-CD）作为手性流动相添加剂,建立了阿卓乳酸对映体的高效液相色谱拆分方法。考察了环糊精衍生物类型、手性添加剂浓度、流动相pH、流速和柱温对手性分离的影响,同时探讨了高效液相色谱采用磺丁基醚-β-环糊精分离阿卓乳酸对映体的分离机制及包结常数,确定了色谱条件为YMC-Pack ODS-A C₁₈色谱柱（250 mm×4.6 mm,5 μm）,流动相为含25 mmol/LSBE-β-CD的0.1 mol/L磷酸盐缓冲液（pH 2.68）-乙腈（90：10,v/v）,流速为0.6 mL/min,柱温为30 ℃,紫外检测波长为220 nm。对映体的保留时间分别为26.65和28.28 min,分离度为1.68。本方法分离度好,简便易行,且比使用手性固定相分离更加经济。     

Optimization of capillary electrophoretic enantioseparation for basic drugs with native beta-CD as a chiral selector

This study presents the advantages of the 20 microm inner diameter (id) capillary for the enantioseparation of ten basic drugs with native beta-CD as the chiral selector. The apparent binding constants of each enantiomeric pair were determined to calculate the optimum beta-CD concentration ([beta-CD]opt) and the optimization was subsequently carried out. Comparison of the 20 microm id with 50 microm id were made in terms of the results obtained in the optimization and detection limits. Applying the optimum conditions for each compound, reproducible results (RSD from 0-3; n>5) were obtained for the 20 microm id capillary. Although the sensitivity is lower in the 20 microm id capillary, the LOD determined using this capillary is still found to be acceptable for the ten basic drugs studied. Enhanced resolution and faster analysis times were the main advantages observed with the use of this capillary in enantioseparation.     

A (4R)-hydroxy-L-proline-derived chiral scaffold and its oligomers as chiral selectors in liquid chromatography chiral stationary phases for enantioseparation

The chromatographic behaviour of a poly-L-proline-derived chiral stationary phase (CSP) is compared to the corresponding single proline-derived CSP. Structurally diverse racemic test compounds and mobile phases, including normal- and RP conditions, were used. Although the application domain of the poly-L-proline-derived CSP (CSP-3) was considerably restricted, this CSP showed a higher retention and a slightly broader application domain than the monomeric analogue (CSP-1) when heptane/2-PrOH was used as mobile phase. The presence of an alcohol in the mobile phase was essential for enantioseparation in the poly-L-proline-derived CSP when normal-phase conditions were applied.     

Enantiomer separation of drugs by capillary electrophoresis using proteins as chiral selectors

The separation of drug enantiomers using proteins as the chiral selectors in capillary electrophoresis (CE) is considered in this review. The proteins used include albumins such as bovine serum albumin, human serum albumin and serum albumins from other species, glycoproteins such as alpha1-acid glycoprotein, crude ovomucoid, ovoglycoprotein, avidin and riboflavin binding protein, enzymes such as fungal cellulase, cellobiohydrolase I, pepsin and lysozyme and other proteins such as casein, human serum transferrin and ovotransferrin. Protein-based CE is carried out in two modes: in one proteins are immobilized or adsorbed within the capillary, or protein-immobilized silica gels are packed into the capillary (affinity capillary electrochromatography mode), and in the other proteins are dissolved in the running buffer (affinity CE mode). Furthermore, the advantages and limitations of the two modes and the factors affecting the chiral separations of various drugs by protein-based CE are discussed.     

Evaluation of cyclodextrin- and cyclofructan-based chiral selectors for the enantioseparation of psychoactive substances in capillary electrophoresis

During this study, a simple and easy-to-prepare electrophoretic method was developed for the enantioseparation of amphetamine and cathinone derivatives. Different types of β-cyclodextrin and cyclofructan-based chiral selectors (CSs), both native and derivatized, were utilized, and the most effective ones, in terms of resolution and analysis time, were identified. In addition, several electrophoretic parameters, such as background electrolyte concentration and pH, and CS concentration, were examined to optimize the separation conditions. Under the optimal electrophoretic conditions, 10 psychoactive substances were enantiomerically separated using 1 mM sulfated cyclofructan-6 (SCF-6) for the amphetamine derivatives and 1 mM sulfated cyclofructan-7 (SCF-7) for the cathinone derivatives dissolved in an aqueous solution of 20-mM monobasic sodium phosphate at pH 2.5, a temperature of 25°C, and an applied voltage of 25 kV. In addition, the method was validated by estimating the intra- and interday precision.     

Comparison of Chirasil-DEX CB as gas chromatographic and ULMO as liquid chromatographic chiral stationary phase for enantioseparation of aryl- and heteroarylcarbinols

For a broad spectrum of simple chiral alcohols, incorporating a (substituted) (het)aryl building block, enantiomer separation characteristics are reported for both gas chromatography on a Chirasil-DEX phase, and liquid chromatography on an (S,S)-ULMO phase. On this chiral Pirkle-type phase, homochiral enantiomers (mostly R) are eluted first without exception. The elution order R before S appears conserved as a rule also for gas chromatographic separations on Chirasil-DEX, though with some remarkable exceptions indicating a change in the dominant discriminative mechanism. This was shown in the homologous series 1-phenylethanol to 1-phenylhexanol having the point of reversal at C4, while the o-methoxy analogues elute from C1 to C4 already in the reversed order.     

High-performance liquid chromatographic enantioseparation of alpha-substituted glycine analogs on a quinine-based anion-exchanger chiral stationary phase under variable temperature conditions

The retention of enantiomers of chiral analytes, i.e. alpha-substituted glycine analogs, on a quinine-based anion-exchanger chiral stationary phase was studied in the temperature range of 5-70 degrees C and at different mobile phase compositions, using isocratic elution in the reversed-phase mode. By variation of both mobile phase composition and temperature, baseline separations could be achieved for these enantiomers. Separation could be optimized more quickly by adjusting the column temperature rather than the mobile phase composition. The dependence of the natural logarithms of retention and selectivity factors (lnk' and lnalpha) on the inverse of temperature, 1/T (van't Hoff plots) was used to determine thermodynamic data on the enantiomers. Calculated thermodynamic constants (Delta(DeltaH degrees ), Delta(DeltaS degrees ) and Delta(DeltaG degrees )) were applied to promote an understanding of the thermodynamic driving forces for retention in this chromatographic system. The elution sequence of the enantiomers in most cases was determined.     

Enantioseparation of novel psychoactive chiral amines and their mixture by capillary electrophoresis using cyclodextrins as chiral selectors

The prevalence of new psychoactive substances (NPS) has been increasing during the last decade as well as their constant growth of availability across the whole world. Regardless of the potential health hazard, NPS (often racemic compounds) are frequently sought after and abused for their psychoactive effects that may differ for individual enantiomers. In this work, capillary electrophoresis was used for the chiral separation of a mixture of eleven psychoactive chiral amines using β-cyclodextrin and carboxymethyl-β-cyclodextrin as chiral selectors at various concentrations. Chiral separation was successful for all the analytes studied. A mixture of these analytes was subsequently analyzed under optimal conditions, i.e., when using 20 mmol/L carboxymethyl-β-cyclodextrin in 50 mmol/L sodium phosphate buffer, pH 2.5. In this case, chiral separation occurred in nine out of eleven analytes. To our best knowledge, we achieved enantioseparations of seven analyzed compounds by CE for the first time.