Enantiomeric separation of FMOC-amino acids by nano-LC and CEC using a new chiral stationary phase, cellulose tris(3-chloro-4-methylphenylcarbamate).

A novel polysaccharide-based chiral stationary phase (CSP), cellulose tris(3-chloro-4-methylphenylcarbamate), also known as Sepapak-2 or Lux Cellulose-2, has been evaluated for the enantiomeric separation of FMOC derivatives of amino acids. After mobile-phase optimization in nano liquid chromatography (nano-LC) the column enabled the enantiomeric separation of 19 out of 23 amino acids tested, indicating the high chiral recognition power of this new CSP. Subsequently, a comparison of the driving force employed (pressure or voltage) was carried out comparing nano-LC and CEC under the same conditions. Better peak efficiencies and resolution were observed by using CEC experiments, which enabled the chiral discrimination of 20 out of 23 amino acids tested. Finally, in order to show the potential of this new CSP, the determination of the content and the enantiomeric purity of the non-protein amino acid citrulline in food supplements was performed. For that purpose, the method was optimized, evaluated and applied to different commercial samples.

Separation of enantiomers of ephedrine by capillary electrophoresis using cyclodextrins as chiral selectors: comparative CE, NMR and high resolution MS studies.

The enantiomer migration order (EMO) of ephedrine was investigated in the presence of various CDs in CE. The molecular mechanisms of chiral recognition were followed for the ephedrine complexes with native α- and ß-CD and heptakis(2,3-di-O-acetyl-6-O-sulfo)-ß-CD (HDAS-ß-CD) by CE, NMR spectroscopy and high-resolution MS. Minor structural differences were observed between the complexes of ephedrine with α- and ß-CD although the migration order of enantiomers was opposite when these two CDs were applied as chiral selectors in CE. The EMO was also opposite between ß-CD and HDAS-ß-CD. Significant structural differences were observed between ephedrine complexes with the native CDs and HDAS-ß-CD. The latter CD was advantageous as chiral CE selector not only due to its opposite electrophoretic mobility compared with that of the cationic chiral analyte, but also primarily due to its enhanced chiral recognition ability towards the enantiomers of ephedrine.

Recent advances in the analysis of antibiotics by CE and CEC.

This article reviews the most recent developments concerning the determination of antibiotics by CE and CEC. The most employed CE separation modes were CZE and MEKC although microemulsion electrokinetic capillary chromatoghraphy was also employed. For the first time, CE was coupled to MS that was applied as a specific and confirmatory detection technique for the analysis of antibiotics. The analytical characteristics of the developed methodologies as well as the different applications reported in the literature on this subject from June 2005 until May 2007 are included in this article. To give the most relevant information on this topic, the experimental conditions employed to achieve the analysis of antibiotics by CE and CEC are provided together with the main applications performed in the pharmaceutical, agrochemical, biological, food, and environmental fields, emphacizing sample preparation requirements needed in each case.

Characterization of protein fractions from Bt-transgenic and non-transgenic maize varieties using perfusion and monolithic RP-HPLC. Maize differentiation by multivariate analysis.

Protein fractions from transgenic Bt and non-transgenic maize varieties, extracted by the Osborne solvent fraction procedure, were characterized for the first time by perfusion and monolithic RP-HPLC in very short analysis times. Albumins and globulins from different transgenic Bt maizes as well as from their non-transgenic isogenic varieties were eluted in four peaks using perfusion RP-HPLC, whereas prolamins and glutelins were separated in seven peaks. Monolithic RP-HPLC enabled the separation of maize proteins in a large number of peaks showing 6 and 10 main peaks for albumins and globulins, respectively. Prolamins migrated at retention times higher than 5 min as seven peaks, whereas glutelins were separated in three main peaks appearing at retention times higher than 6.0 min. Moreover, chromatograms of the whole protein extracts showed 8 and 11 components for perfusion and monolithic RP-HPLC, respectively. A comparison of the chromatograms of the whole protein extracts relative to transgenic and non-transgenic varieties evidenced quantitative differences on the percentages of area, mainly for peaks 2 and 3 by perfusion RP-HPLC and for peaks 3 and 7 by monolithic RP-HPLC. A discriminant analysis based on these proteic profiles was carried out to classify and predict transgenic Bt maize lines, achieving 100% correct classification using perfusion RP-HPLC.

Rapid determination of salbutamol in pharmaceutical preparations by chiral capillary electrophoresis.

A fast and simple method of chiral capillary electrophoresis (CE) has been applied to the analysis of salbutamol in different pharmaceutical preparations. Using of a 25 mM acetate buffer (pH 5.0), containing 13.1 mg/mL carboxymethyl-beta-cyclodextrin (CM-beta-CD), an applied voltage of 20 kV and a temperature of 25 degrees C, the enantiomers of salbutamol could be separated in about 2 min. Three different pharmaceutical preparations (two syrups, one oral solution, and two kind of tablets) containing a racemate of salbutamol were injected directly in the CE system, following dilution in dimethyl sulfoxide (DMSO). Appreciable differences in the retention times were observed for salbutamol enantiomers in the different formulations studied, which were attributed to the effect of the matrix components on the electrophoretic mobility. The standard addition method was used for the calibration due to the existence of matrix interferences. Finally, the stability of the enantiomers of salbutamol in the oral solution was studied calculating the enantiomeric ratio values when the solution was injected immediately after being opened in the first case and after being opened and stored in the fridge for two months in the second case.

Retention modeling and resolution optimization for a group of N-phenylpyrazole derivatives in micellar electrokinetic chromatography using empirical and physicochemical models.

The optimization of the separation resolution for a group of N-phenylpyrazole derivatives in micellar electrokinetic chromatography (MEKC) as a function of the separation buffer composition (surfactant and organic modifier concentration) has been performed. In order to achieve our purpose, the first step has been the prediction of the migration times of the electroosmotic flow (t(0)) and micelles (t(m)), and the retention factors of solutes (k), as a function of surfactant (sodium dodecyl sulfate) and alcohol (n-propanol or n-butanol) concentrations, by means of empirical equations. Also, some physicochemical models have been applied to relate the retention factors to the surfactant and the organic modifier concentrations in order to optimize the separation resolution and to increase our knowledge of the separation process. Finally, a comparison of the resolution optimization through the use of the physicochemical and empirical models selected has been made in order to obtain the optimum separation buffer composition for the separation of a group of 17 N-phenylpyrazole derivatives as test solutes.