Enantiomeric resolution by HPLC of axial chiral amides using amylose tris[(S)-1-phenylethylcarbamate]

The enantiomeric resolution of a series of N-arylamides was examined on amylose tris[(S)-1-phenylethylcarbamate] coated onto aminopropylated 7 μm silica with 500 Å diameter pores and on naked silica 5 μm particle size with 500 Å diameter pores. The enantiomeric resolution obtained for this series was excellent on both columns. The enantioselectivity of cellulose and amylose tris (3,5-dimethylphenylcarbamate) coated onto APS-Hypersil (120 Å pore size, 5 μm particle size) was also investigated for this series of compounds. Chirality 9:109–112, 1997. © 1997 Wiley-Liss, Inc.     

Chiral separation performance of micrometer‐sized monodispersed silica spheres with high protein loading

Micrometer‐sized monodispersed silica spheres (～360 μm) with high loading of bovine serum albumin (BSA) (～450 mg/g) were prepared as an adsorbent for large‐scale chiral separation. The new adsorbent was characterized with scanning electron microscopy (SEM), nitrogen adsorption‐desorption isotherms, the mercury intrusion method, infrared spectroscopic analysis, and elementary analysis. The extent of chiral separation was tested with rac‐tryptophan (rac‐Trp) and rac‐phenylalanine (rac‐Phe) as solutes. The results showed that the absorbent exhibited a high surface area, large pore volume, and bimodal macromesoporous structures, enabling fast mass transfer and high separation efficiency. A fast adsorption rate, reaching equilibrium in less than 6 min, and a high degree of chiral separation, with the enantiomer excess (e.e.) value reaching as high as 100% in the first 10 min, was observed in a small (5 cm in height, 0.46 cm in internal diameter) packed column that could be regenerated with a pH 5.0 HAc‐NaAc buffer. The results show that monodispersed silica spheres with a high BSA loading have great potential for applications in large‐scale chiral separation processes. Chirality, 2009. © 2008 Wiley‐Liss, Inc.     

Determination of carbamazepine in human urine and serum samples by high‐performance liquid chromatography with post‐column Ru(bipy)‐Ce(SO4)2 chemiluminescence detection

A novel, sensitive and rapid CL method coupled with high‐performance liquid chromatography separation for the determination of carbamazepine is described. The method was based on the fact that carbamazepine could significantly enhance the chemiluminescence of the reaction of cerium sulfate and tris(2,2‐bipyridyl) ruthenium(II) in the presence of acid. The chromatographic separation was performed on a Kromasil® (Sigma‐Aldrich) TM RP‐C18 column (id: 150 mm × 4.6 mm, particle size: 5 µm, pore size: 100 Å) with a mobile phase consisting of methanol–water‐glacial acetic acid (70:29:1, v/v/v) at a flowrate of 1.0 mL/min, the total analysis time was within 650 s. Under optimal conditions, CL intensity was linear for carbamazepine in the range 2.0 × 10^?8 ~ 4.0 × 10^?5 g/mL, with a detection limit of 6.0 × 10^?9 g/mL (S/N = 3) and the relative standard detection was 2.5% for 2.0 × 10^?6 g/mL (n = 11). This method was successfully applied to the analysis of carbamazepine in human urine and serum samples. The possible mechanism of the CL reaction is also discussed briefly. Copyright © 2012 John Wiley & Sons, Ltd.     

Variables in the high-performance anion-exchange chromatography of proteins

The effect of mobile phase velocity, separation time, support pore diameter, column length, and temperature on resolution and loading capacity of a new commercially available high-performance anion-exchange support, SynChropak AX-300, has been examined. This material is a macroporous spherical silica of 10 μm particle size with a bonded polymeric amine layer. It was found that the heterogeneity of ovalbumin samples, combined with bovine serum albumin, make them useful probes in evaluation of anion-exchange supports. In the columns of 4.1 mm i.d., the highest resolutions of proteins were achieved at a flow rate of 0.25 ml/min. Up to 10 mg of protein per injection could be applied on a 4.1 × 250 mm AX-300 column with good resolution. Columns of 50 mm length had one-tenth the protein load capacity of a 250-mm column, retaining approximately 75% of the resolution.     

腾格里沙漠草方格固沙林土壤颗粒组成、分形维数及其对土壤性质的影响

为了明确草方格人工固沙造林植被恢复过程中土壤颗粒组成、分形维数及对土壤理化性质的影响,以腾格里沙漠东南缘2016年(1 a)、2013年(4 a)和1987年(30 a)草方格固沙林为研究样地,以周围流动沙地为对照(CK),研究了草方格固沙造林后植被恢复过程中土壤颗粒组成、分形维数及与土壤理化性质的作用关系.结果表明: 100～250、250～500 μm土壤颗粒含量较高,分别为42.5%～80.1%、12.5%～42.2%;50～100 μm土壤颗粒含量居中,为0.2%～20.8%;<2和2～50 μm的土壤颗粒含量次之,分别在0～1.3%和0～22.7%;而500～1000 μm的土壤颗粒含量较低,在0.3%以下.<2和2～50 μm土壤颗粒仅在30 a固沙林有分布;50～100 μm土壤颗粒分布为30 a最高,4 a和1 a居中,而CK最低;100～250 μm土壤颗粒分布依次为4 a>1 a>CK>30 a;250～500 μm土壤颗粒分布为CK>1 a>4 a>30 a;但500～1000 μm土壤颗粒在各样地分布均较少,且不同样地之间无显著差异.研究区土壤颗粒分形维数为0.54～2.59,并且不同样地间存在显著差异,表现为30 a最高,4 a与1 a居中,而CK最低.土壤颗粒分形维数与土壤黏粒、粉粒、极细砂粒含量呈极显著正相关,而与土壤中砂粒呈极显著负相关.土壤颗粒分形维数与土壤电导率、有机碳、全氮和碳氮比均呈极显著正相关,而与土壤pH和含水量无相关性.土壤中<2、2～50、50～100 μm颗粒与土壤电导率、有机碳、全氮和碳氮比均呈极显著正相关,而250～500 μm土壤颗粒与上述4个土壤指标和土壤含水量呈显著负相关.500～1000 μm土壤颗粒与土壤含水量亦呈极显著负相关.在腾格里沙漠东南缘地区利用草方格进行人工固沙植被建设,可有效促进土壤颗粒细粒化,长期演变导致土壤黏粒和粉粒及土壤分形维数显著增加,促使土壤有机碳和全氮含量提高,有利于土壤理化性质改善和促进沙漠化治理.     

Development of a validated capillary electrophoresis method for enantiomeric purity control and quality control of levocetirizine in a pharmaceutical formulation

A chiral capillary electrophoresis method has been developed for the quantification of 0.1% of the enantiomeric impurity (dextrocetirizine) in levocetirizine and determination of both in pharmaceuticals using sulfated-β-cyclodextrins (CDs) as chiral selector. Several parameters affecting the separation were studied such as the type and concentration of chiral selectors, buffer composition and pH, organic modifier, mixtures of two CDs in a dual system, voltage, and temperature. The optimal separation conditions were obtained using a 50 mM tetraborate buffer (pH 8.2) containing 1% (w/v) sulfated-β-CDs on a fused-silica capillary. Under these conditions, the resolution of two enantiomers was higher than 3. To validate the method, the stability of the solutions, robustness (two level half fraction factorial design for 5 factors using 19 experiments [2(n-1)+3]), precision, linearity (dextrocetirizine 0.25-2.5 μg/ml, R(2) = 0.9994, y = 0.0375x + 0.0008; levocetirizine 15-100 μg/ml, R(2) = 0.9996, y = 0.0213x + 0.0339), limit of detection (0.075 μg/ml, 0.03% m/m), limit of quantification (0.25 μg/ml, 0.1% m/m), accuracy (dextrocetirizine 84-109%, levocetirizine 97.3-103.1%), filter effect, and different CD batches were examined. The validated method was further applied to bulk drug and tablets of levocetirizine.     

Bioanalytical chiral chromatographic technique and docking studies for enantioselective separation of meclizine hydrochloride: Application to pharmacokinetic study in rabbits

Enantiomeric resolution and molecular docking studies of meclizine hydrochloride on polysaccharide-based chiral stationary phase comprising cellulose tris(4-methylbenzoate) chiral selector (150 × 4.6 mm, 3.0 μm) were presented. The mobile phase used was acetonitrile:10mM ammonium bicarbonate (95:05, v/v). The developed technique was used to perform the enantioselective assay of meclizine hydrochloride in its marketed formulation. The elution order of meclizine hydrochloride enantiomers was determined by docking studies. Target compound was extracted from rabbit plasma using protein precipitation technique, followed by development of bioanalytical chiral separation method using the same matrix. Application of the method to determine pharmacokinetic parameters of meclizine hydrochloride enantiomers was performed using Phoenix WinNonlin 8.1 software. The results demonstrated stereoselective disposition of meclizine hydrochloride enantiomers in rabbits.     

Enantioselective potential of polysaccharide‐based chiral stationary phases in supercritical fluid chromatography

The enantioselective potential of two polysaccharide‐based chiral stationary phases for analysis of chiral structurally diverse biologically active compounds was evaluated in supercritical fluid chromatography using a set of 52 analytes. The chiral selectors immobilized on 2.5 μm silica particles were tris‐(3,5‐dimethylphenylcarmabate) derivatives of cellulose or amylose. The influence of the polysaccharide backbone, different organic modifiers, and different mobile phase additives on retention and enantioseparation was monitored. Conditions for fast baseline enantioseparation were found for the majority of the compounds. The success rate of baseline and partial enantioseparation with cellulose‐based chiral stationary phase was 51.9% and 15.4%, respectively. Using amylose‐based chiral stationary phase we obtained 76.9% of baseline enantioseparations and 9.6% of partial enantioseparations of the tested compounds. The best results on cellulose‐based chiral stationary phase were achieved particularly with propane‐2‐ol and a mixture of isopropylamine and trifluoroacetic acid as organic modifier and additive to CO₂, respectively. Methanol and basic additive isopropylamine were preferred on amylose‐based chiral stationary phase. The complementary enantioselectivity of the cellulose‐ and amylose‐based chiral stationary phases allows separation of the majority of the tested structurally different compounds. Separation systems were found to be directly applicable for analyses of biologically active compounds of interest.     

Intraparticle concentration gradients for substrate and acidic product in immobilized cephalosporin C amidase and their dependencies on carrier characteristics and reaction parameters

Cephalosporin C amidase was covalently attached using a protein loading of 7.0–200 mg protein/g dry carrier on four epoxy‐activated Sepabeads differing in particle size and pore diameter. Initial‐rate kinetic analysis showed that for Sepabeads with small pore diameters (30–40 nm), the apparent K_M of the amidase for hydrolysis of cephalosporin C at 37°C and pH 8.0 increased ～3‐fold in response to increased particle size (～120–400 µm) and increased amount of immobilized enzyme (7.0–70 mg protein/g dry carrier) while maximum specific activity (3.2 U/mg protein; 25% of free amidase) was affected only by particle size. In contrast, for Sepabeads with wide pores (150–250 nm), the K_M was independent of the enzyme loading. Internal effectiveness factors calculated from observable Thiele modulus reflected the dependence of K_M on geometrical parameters of the particles. A new method for determination of the overall intraparticle pH was developed based on luminescence lifetime measurements in the frequency domain. Sepabeads were doubly labeled using a lipophilic variant of the pH‐sensitive dye fluorescein, and Ru(II) tris(4,7‐diphenyl‐1,10‐phenantroline) whose phosphorescence properties are independent of pH. Luminescent lifetime measurements of doubly labeled particle suspensions showed superior signal‐to‐noise ratio compared to fluorescence intensity‐based measurements using singly labeled particles. The difference at apparent steady state (ΔpH) between bulk (external pH) and intraparticle pH (internal pH) was as large as ～0.6 units. The ΔpH was dependent on substrate concentration, particle size, and pore diameter. Therefore, these results characterize the role of carrier characteristics and reaction parameters in the formation of concentration gradients for substrate and acidic product during hydrolysis of cephalosporin C by immobilized amidase. The strong pH dependence of the immobilized amidase underscores the importance of considering intraparticle pH gradients in the design of an efficient carrier‐bound biocatalyst. Biotechnol. Bioeng. 2010;106: 528–540. © 2010 Wiley Periodicals, Inc.     

Change in the concentrations of iron in different size fractions during a phytoplankton bloom in controlled ecosystem enclosures

To observe micronutrient dynamics in the plankton ecosystem, controlled ecosystem enclosure (CEE) experiments were conducted in Saanich Inlet, B.C., Canada. Two CEEs (2.5 m in diameter, 16 m in length, one for Fe studies and the other for biological studies) were launched for the period 22 July to 5 August 1996 and enriched with 10 μM nitrate and 5.2 nM Fe (13% of total Fe) on day 1. Sampling from three integrated depths, intervals 0-4, 4-8 and 8-12 m, was performed on days 0, 1, 2, 3, 4, 5, 7, 9 and 11. Iron concentrations were measured for five size fractions: >25 μm particles, 2-25 μm particles, 0.2-2 μm particles, 0.2 μm-200 kDa small colloidal particles and <200 kDa soluble species. The sediment in the Fe enclosure was also collected on every sampling day after day 2 and its Fe was determined. Size-fractionated particulate organic carbon and total chlorophyll-a were also analyzed.The Fe in small colloidal particles (200 kDa-0.2 μm) comprised 78% of the traditionally defined dissolved phase (<0.2 μm) on day 1. Of all the size fractions of Fe, the small colloidal particulate fraction decreased most significantly during the phytoplankton bloom. In the dissolved fraction (<0.2 μm), the small colloidal particle fraction comprised 79% of the decrease. The decrease in concentration of Fe in small colloidal particles was larger than that of total Fe from day 1 to day 4. In contrast, the >25 μm Fe particles increased over the same period. These results suggest that Fe in small colloidal particles changed to >25 μm Fe particles during phytoplankton growth. A large amount of Fe was kept in the surface layer with the phytoplankton, and transported to the deep layer by phytoplankton sedimentation, at the end of the bloom. From these results, the small colloidal particulate Fe seems to be the most dynamic size fraction and a high percentage of Fe in small colloidal particles changed to large particles due to chemical/physical aggregation and/or physical adsorption to suspended particles such as phytoplankton cells.     

Theoretical analysis of chromatographic peak asymmetry and sharpness by the moment method using two peptides

The analyses of peak shapes in chromatography are useful in operating chromatographic system. The asymmetry and sharpness of a chromatographic peak are estimated by the reversed-phase adsorption of two standard peptides (angiotensin II bradykinin) on C₁₈. In this work, the average particle diameters of C₁₈ were 5 and 15 μm, while the pore sizes were 100 and 300 Å. The composition of the mobile phase was 50/50 vol. % of a binary mixture of acetonitrile and water with 0.1% TFA, and the particles were packed in a stainless column (4.6×150 mm). The third and the fourth central movement were calculated from the chromatographic elution curves by moment analysis. The peak asymmetry was determined by two theoretical calculations: the asymmetry factor by elution peak analysis and skewness with moment analysis. The sharpness was estimated by the fourth central moment. In this work, the most acceptable skewness was calculated when the pore size was 300 Å. The large excess was observed on small pore size.     

Methane recovery from water hyacinth through whole-cell immobilization technology

The concepts of feed pretreatment, phase separation, and whole-cell immobilization technology have been incorporated in this investigation for the development of rational and cost-effective two- and three-stage methane recovery systems from water hyacinth (WH)Analyses of laboratory data reveal that a three-stage system could be designed with an alkali pretreatment stage [3.6% Na(2)CO(3) + 2.5% Ca(OH)(2) W/W, 24 h HRT] followed by an open acid reactor (2.1 days HRT) and closed immobilized methane reactor (12 h HRT), providing steady-state COD conversion of 62-65%, TVA conversion of 91-95%, and gas productivity of 4.08-5.36 L/L reactor volume/day with 82% methane. A gas yield of 50 L/kg WH/day (dry wt basis) at 35-37 degrees C is possible with this system. Insulation bricks, with particle size distribution of 500-3000 mum, were used as support material in the reactors at organic loading rate of 20 kg COD/m(3) day. The reactors matured in 15-18 weeksSubstantial reduction in retention time for the conversion of volatile acids in immobilized methane reactors prompted further research on the combined immobilized reactor to make possible an additional reduction in the cost of a WH-based biogas system. Evaluation of laboratory data reveals that a two-stage system could be designed with an open alkali pretreatment stage and a combined immobilized reactor (12 h HRT), providing steady-state COD conversion of 53% and gas productivity of 3.1 L/L reactor volume/day with 86% methane. A gas yield of 44 L/kg WH/day (dry wt basis) at 35-37 degrees C could be obtained from this system. Insulation bricks, with 500-1000 mum particle size distribution, was used as support material at an organic loading rate of 15 kg COD/m(3) day. Notwithstanding the fact that the technology in this study has been developed with water hyacinth as substrate, the implicit principles could be extended to any other organic substrate.     

Enantioseparations with cellulose tris(3-chloro-4-methylphenylcarbamate) in nano-liquid chromatography and capillary electrochromatography

Cellulose tris(3-chloro-4-methylphenylcarbamate) was coated onto native and aminopropylsilanized silica in order to prepare chiral stationary phases (CSPs) for enantioseparations using nano-liquid chromatography (nano-LC) and capillary electrochromatography (CEC). The effect of the chiral selector loading onto silica, mobile phase composition and pH, as well as separation variables on separation of enantiomers was studied. It was found that CSPs based on cellulose tris(3-chloro-4-methylphenylcarbamate) can be used for preparation of very stable capillary columns useful for enantioseparations in nano-LC and CEC in combination with polar organic mobile phases.     

Liquid chromatographic separation and thermodynamic investigation of lorcaserin hydrochloride enantiomers on immobilized amylose–based chiral stationary phase

A novel liquid chromatographic method was developed for enantiomeric separation of lorcaserin hydrochloride on Chiralpak IA column containing chiral stationary phase immobilized with amylose tris (3.5‐dimethylphenylcarbamate) as chiral selector. Baseline separation with resolution greater than 4 was achieved using mobile phase containing mixture of n‐hexane/ethanol/methanol/diethylamine (95:2.5:2.5:0.1, v/v/v/v) at a flow rate of 1.2 mL/min. The limit of detection and limit of quantification of the S‐enantiomer were found to be 0.45 and 1.5 μg/mL, respectively; the developed method was validated as per ICH guideline. The influence of column oven temperatures studied in the range of 20°C to 50°C on separation was studied; from this, retention, separation, and resolution were investigated. The thermodynamic parameters ΔH°, ΔS°, and ΔG° were evaluated from van't Hoff plots,(Ink′ _versus 1/T) and used to explain the strength of interaction between enantiomers and immobilized amylose–based chiral stationary phase     

Cross-linked glucose isomerase crystals as a liquid chromatographic separation material

Cross-linked crystals of glucose isomerase (CLGI) were characterized as a liquid chromatographic separation material. The experiments were done with crystals having an average diameter of 83 μm. Porosity (epsilon(p)) and pore size distribution of the CLGI crystals were measured with size exclusion chromatography using D(2)O and polyethylene glycols as probes. CLGI material was capable of separating <1000 g/mol polyethylene glycols. Fifty two percent (epsilon(p) = 0.47) of the total crystal volume was in pores. Pore size measurement showed that CLGI crystals were microporous material, having an average apparent pore diameter of 29 +/- 0.08 ?. CLGI material separated n-alcohols C(1) to C(8) based on the hydrophobic interaction between the protein material and the carbon chain of the alcohols. Height equivalent to a theoretical plate (HETP, in millimeters) ranged from 1.6 to 0.89 for the C(1) to C(7) n-alcohol series. Despite the large crystal size, CLGI as a chirally active phase effectively separated D- and L-arabitol (R(s) = 0.58) and showed potential for chiral separation of amino acids.     

Formulation optimization of chelerythrine loaded O-carboxymethylchitosan microspheres using response surface methodology

The aims of this investigation were to develop a procedure to prepare chelerythrine (CHE) loaded O-carboxymethylchitosan (O-CMCS) microspheres by emulsion cross-linking method and optimize the process and formulation variables using response surface methodology (RSM) with a three-level, three-factor Box-Behnken design (BBD). The independent variables studied were O-CMCS/CHE ratio, O/W phase ratio, and O-CMCS concentration, dependent variables (responses) were drug loading content and encapsulation efficiency. Mathematical equations and response surface plots were used to relate the dependent and independent variables. The process and formulation variables were optimized to achieve maximum drug loading content and entrapment efficiency by the desirability function. The optimized microsphere formulation was characterized for particle size, shape, morphology and in vitro drug release. Results for mean particle size, drug loading content, entrapment efficiency, and in vitro drug release of CHE-loaded O-CMCS microspheres were found to be of 12.18 μm, 4.16 ± 3.36%, 57.40 ± 2.30%, and 54.5% at pH 7.4 after 70 h, respectively. The combination use of RSM, BBD and desirability function could provide a promising application for O-CMCS as controlled drug delivery carrier and help to develop procedures for a lab-scale microemulsion process.     

Modeling flux in tangential flow filtration using a reverse asymmetric membrane for Chinese hamster ovary cell clarification

Tangential flow filtration is advantageous for bioreactor clarification as the permeate stream could be introduced directly to the subsequent product capture step. However, membrane fouling coupled with high product rejection has limited its use. Here, the performance of a reverse asymmetric hollow fiber membrane where the more open pore structure faces the feed stream and the barrier layer faces the permeate stream has been investigated. The open surface contains pores up to 40 μm in diameter while the tighter barrier layer has an average pore size of 0.4 μm. Filtration of Chinese hamster ovary cell feed streams has been investigated under conditions that could be expected in fed batch operations. The performance of the reverse asymmetric membrane is compared to that of symmetric hollow fiber membranes with nominal pore sizes of 0.2 and 0.65 μm. Laser scanning confocal microscopy was used to observe the locations of particle entrapment. The throughput of the reverse asymmetric membrane is significantly greater than the symmetric membranes. The membrane stabilizes an internal high permeability cake that acts like a depth filter. This stabilized cake can remove particulate matter that would foul the barrier layer if it faced the feed stream. An empirical model has been developed to describe the variation of flux and transmembrane pressure drop during filtration using reverse asymmetric membranes. Our results suggest that using a reverse asymmetric membrane could avoid severe flux decline associated with fouling of the barrier layer during bioreactor clarification.     

Chromatographic enantioseparation by cycloalkylcarbamate derivatives of cellulose and amylose

Cyclopentyl and (+/-)-exo-2-norbornylcarbamates of cellulose and amylose were prepared and their chiral recognition abilities as chiral stationary phases for high-performance liquid chromatography (HPLC) were evaluated. Among these carbamates, cellulose tris(cyclopentylcarbamate) and amylose tris((+/-)-exo-2-norbornylcarbamate) showed particularly high chiral recognition, which is comparable to that of several well-known phenylcarbamate derivatives. The chiral recognition mechanism of cellulose tris(cyclohexylcarbamate), which was previously found to be an effective chiral stationary phase for HPLC, was investigated using NMR spectroscopy. The derivative dissolved in chloroform exhibited the chiral discrimination of several enantiomers in NMR as well as in HPLC. For example, the 1,1'-bi-2-naphthol enantiomers were distinctly discriminated in the (1)H, (13)C, and 2D-NOESY spectra.     

Forensic analysis of eleven cyclic antidepressants in human biological samples using a new reversed-phase chromatographic column of 2 μm porous microspherical silica gel

A high-performance liquid chromatographic method has been developed for the forensic analysis of eleven frequently used cyclic antidepressant drugs (ADSs) (amitriptyline, amoxapine, clomipramine, desipramine, dosulepine, doxepin, imipramine, maprotiline, melitracen, mianserine and nortriptyline) using a recently developed reversed-phase column with 2 μm particles for the analysis of biological samples. The separation was carried out using two different C₈ reversed-phase columns (column 1: 100 mm × 4.6 mm I.D., particle size 2 μm, TSK gel Super-Octyl; column 2: 100 mm × 4.6 mm I.D., particle size 5 μm, Hypersil MOS-C₈) for comparison. The mobile phase was composed of methanol-20 mM KH₂PO₄ (pH 7) (60:40, v/v) and the flow-rate was 0.6 ml/min for both columns. The absorbance of the eluent was monitored at 254 nm. When the eleven drugs were determined, the sensitivity with the 2 μm particles was about five times greater than with the 5 μm particles. Retention times on column 1 were shorter than those on column 2. These results show that the new ODS column packing with a particle size of 2 μm gives higher sensitivity and a shorter analysis time than the conventional ODS column packing when applied to the analysis of biological samples.     

Chiral separation of cathinone derivatives used as recreational drugs by HPLC-UV using a CHIRALPAK® AS-H column as stationary phase

Cathinone derivatives gained high popularity on the recreational drugs market during the past 10 years. All these compounds are chiral, and the pharmacological potency of the enantiomers of these stimulants is supposed to differ. The goal of this research was to develop a reliable and easy‐to‐perform high‐performance liquid chromatography ultraviolet method for the chiral separation of a set of 24 cathinone derivatives. A commercially available CHIRALPAK® AS‐H column consisting of amylose tris [(S)‐α‐methylbenzylcarbamate] coated on 5‐µm silica gel was found to be suitable to resolve a majority of the tested compounds. High‐performance liquid chromatography measurements were performed in normal phase mode under isocratic conditions with a mobile phase consisting of hexane, isopropanol, and triethylamine at a flowrate of 1 ml/min. The ratio between hexane and isopropanol was optimized by means of three model substances. Under final conditions with a mobile phase of hexane, isopropanol, and triethylamine (97:3:0.1), 19 out of 24 compounds were successfully resolved into their enantiomers and detected at a wavelength of 254 nm. A correlation between the substituents of the nitrogen atom and the separation results are shown. Furthermore, enantiomer separation results of four cathinone derivatives were compared with the results of their amphetamine analogs. Chirality 24:486–492, 2012. © 2012 Wiley Periodicals, Inc.