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P Jandera

Publications and source records attributed to P Jandera.

At least 19 recordsLinked to original sources

Monolithic continuous beds as a new generation of stationary phase for chromatographic and electro-driven separations.

Monolithic supports are a new generation of stationary phases which are typically prepared using a simple molding process carried out within the confines of the capillary. They provide high rates of mass transfer at lower pressure drops, enable much faster separations and the nature of the pores allows easy permeability for large molecules. This review summarizes the current achievements and application of organic polymer-based monolithic columns, silica-based monolithic columns and their application in bioaffinity processing, modern biotechnology, determination of microorganisms and chiral separations. Special attention is paid to microfabricated devices with monolithic supports because their fabrication of particles directly in the channel eliminates the need for a frit and also creates a unique homogeneity of packing.

Chromatography, Liquid↗

Effects of ion-pairing reagents on the electrospray signal suppression of sulphonated dyes and intermediates.

High-performance liquid chromatography/mass spectrometry (HPLC/MS) analysis of anionic species such as sulphonic acid dyes and intermediates requires volatile ion-pairing mobile phase additives. Six di- and trialkylammonium acetates were compared with tetraalkylammonium salts and ammonium acetate in the concentration range 0-20 mmol l(-1) as mobile phase additives for HPLC/MS of polysulphonated compounds. The effects of the structure and concentration of the ion-pairing reagents on the electrospray response of mono-, di- and tetrasulphonic aromatic acids and acid dyes were studied in detail. Further, five different mass analysers and instrument geometries were compared. A higher signal decrease is observed with linear geometry instruments in comparison to orthogonal or even Z-spray geometry mass spectrometers. The concentration of mobile phase additives has a significant influence on the abundance ratios of multiply charged ions in the mass spectra of polysulphonated compounds. The competing ions of sulphonic acids may also cause significant signal suppression.

Journal Article↗

Effects of substituted cyclodextrins on the separation of aromatic sulphonic acids by capillary zone electrophoresis.

Effects of the addition of various additives in the working electrolyte on the selectivity of capillary electrophoretic separation of naphthalenesulphonic acids used as intermediates in the production of synthetic dyes were investigated. Cyclodextrins form inclusion complexes with various compounds and are not only excellent chiral selectors, but have been also successfully applied for separations of positional isomers. In this work, methyl-beta-cyclodextrin, heptakis(2,6-di-O-methyl)-beta-cyclodextrin, heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin, (2-hydroxypropyl)-beta-cyclodextrin and (2-hydroxypropyl)-gamma-cyclodextrin were studied as isomeric selector additives and compared with unsubstituted beta-cyclodextrin. In addition to the size of the cyclodextrin cavity, the number and type of the substituents in the cyclodextrin molecules strongly affect the separation of isomeric naphthalenesulphonic acids, but the effect of the substituted cyclodextrins on the separation selectivity is different for various types of sulphonic acids. Best separations of non-substituted naphthalenesulphonic acids were achieved in a borate buffer with methyl-beta-cyclodextrin, whereas the running buffer with non-substituted beta-cyclodextrin provides superior separation of amino and hydroxynaphthalenesulphonic acids.

2-Hydroxypropyl-beta-cyclodextrin↗

High performance liquid chromatography-mass spectrometric analysis of sulphonated dyes and intermediates.

This paper reports our results in the analysis of polysulphonated anionic dyes and their intermediates using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Negative-ion electrospray ionization is the most suitable ionization technique for the molecular mass determination of polysulphonated dyes or other dyes carrying a negative charge. From the series of [M-xH]x- ions and their sodiated adducts [M-(x + y)H+yNa]x-, the molecular mass and the number of sulphonic and carboxylic groups can be determined. The mobile phase should be compatible with the mass spectrometric detection, which rules out non-volatile tetraalkylammonium salts usually used as ion-pair mobile phase additives for the HPLC of sulphonated compounds. Some mono- and disulphonated dyes and intermediates can be separated with aqueous-organic mobile phases containing 5 mM ammonium acetate, which is the most suitable additive as far as compatibility with MS detection is concerned. However, the retention of compounds with two or more sulphonic groups is too low for a successful separation both with this mobile phase additive and with ion-pair additives with short alkyl chains. The dihexylammonium acetate ion-pairing reagent offers a reasonable compromise in terms of sufficient volatility and adequate retention and separation selectivity for the HPLC-MS analysis of polysulphonated dyes.

Chromatography, High Pressure Liquid↗

Fitting adsorption isotherms to the distribution data determined using packed micro-columns for high-performance liquid chromatography.

Knowing the adsorption isotherms of the components of a mixture on the chromatographic system used to separate them is necessary for a better understanding of the separation process and for the optimization of the production rate and costs in preparative high-performance liquid chromatography (HPLC). Currently, adsorption isotherms are usually measured by frontal analysis, using conventional analytical columns. Unfortunately, this approach requires relatively large quantities of pure compounds, and hence is expensive, especially in the case of pure enantiomers. In this work, we investigated the possible use of packed micro-bore and capillary HPLC columns for the determination of adsorption isotherms of benzophenone, o-cresol and phenol in reversed-phase systems and of the enantiomers of mandelic acid on a Teicoplanin chiral stationary phase. We found a reasonable agreement between the isotherm coefficients of the model compounds determined on micro-columns and on conventional analytical columns packed with the same material. Both frontal analysis and perturbation techniques could be used for this determination. The consumption of pure compounds needed to determine the isotherms decreases proportionally to the second power of the decrease in the column inner diameter, i.e. 10 times for a micro-bore column (1 mm I.D.) and 100 times for capillary columns (0.32 mm I.D.) with respect to 3.3 mm I.D. conventional columns.

Adsorption↗

Effect of the mobile phase on the retention behaviour of optical isomers of carboxylic acids and amino acids in liquid chromatography on bonded Teicoplanin columns.

Conditions for separation of enantiomers of underivatized amino acids phenyl glycine and tryptophan and of mandelic acid as test compounds were studied on a Chirobiotic T column packed with amphoteric glycopeptide Teicoplanin covalently bonded to the surface of silica gel. The effects of the mobile phase composition on the retention and selectivity under analytical conditions, on the profile of the adsorption isotherms of the enantiomers and on the overloaded separation were investigated. The concentration of ethanol or of methanol in aqueous-organic mobile phases and the pH of the mobile phase affect not only the retention and selectivity, the saturation capacity and the isotherm profile, but also the solubility of the acids, which should be taken into account in development of preparative separations. A compromise between the separation selectivity and the solubility should be made in selecting the mobile phase suitable to accomplish preparative separations at acceptable production rate and throughput of the operation.

Adsorption↗

Characterisation of retention in micellar high-performance liquid chromatography, in micellar electrokinetic chromatography and in micellar electrokinetic chromatography with reduced flow.

The retention (migration) behaviour of various barbiturates, phenylurea and triazine herbicides in micellar electrokinetic chromatography (MEKC) with uncoated fused-silica capillaries was compared with the behaviour in micellar electrokinetic chromatography with reduced electroosmotic flow (RF-MEKC) using capillaries modified with linear polyacrylamide. The error in the values of the retention factors caused by the neglection of the contribution of the electroosmotic flow in RF-MEKC was investigated and a method for correcting this error was suggested. The retention was characterised using the lipophilic and polar indices to characterise and to predict the retention as a function of the concentration of the surfactant (sodium dodecylsulphate) in the running buffer in MEKC and in RF-MEKC. Homologous series of n-alkylbenzenes and of n-alkan-2-ones were compared as the standard sets for the calibration of the retention (migration) index scale. The values of the lipophilic indices of a given solute measured in reversed-phase HPLC, MEKC and RF-MEKC are close to each other. Under ideal MEKC conditions, the values of the polarity indices are close to one for various sample solutes. However, for partially ionised compounds such as weakly acidic barbiturates, where the contribution of the electrophoretic migration is significant, the values of the polarity indices are significantly lower than one. Optimum conditions for separations of mixtures of triazine and phenylurea herbicides and of barbiturates using various techniques tested were compared.

Chromatography, High Pressure Liquid↗

Description of adsorption equilibria in liquid chromatography systems with binary mobile phases.

Adsorption of some simple compounds from pure and mixed solvents and of some solvents from mixed binary solvent mixtures on columns used in normal-phase, aqueous-organic and non-aqueous liquid chromatography was investigated. The distribution of the compounds between the liquid and the stationary phase is affected by the composition of the solvent mixture. Although preferential sorption of stronger solvents can often be described by the Langmuir isotherm, significant deviations are observed in some systems. A model was suggested accounting for the deviations from Langmuir isotherm by association on already adsorbed molecules. In most systems studied in this work, a simple competitive Langmuir isotherm did not provide a good fit to the experimental distribution data from mixed solvents when competition between the solute and the strong solvent for the adsorption sites was considered. A competitive isotherm taking into account possible association of solute on its own already adsorbed molecules and on the adsorbed molecules of strong solvent improves the fit to the experimental distribution data and enables description of the distribution data in dependence on the concentration of the strong solvent in mixed solvents in normal-phase, aqueous-organic and non-aqueous reversed-phase systems. Even though we need more data to prove the validity of the present model, we observe a good qualitative agreement of the experimental isotherm coefficients with the relative strength of the interactions expected in various chromatographic systems employing mixed solvents.

Adsorption↗

Study of the adsorption behavior of the enantiomers of 1-phenyl-1-propanol on a cellulose-based chiral stationary phase.

Using single-step frontal analysis, we measured single-component and competitive adsorption isotherm data for the two enantiomers of 1-phenyl-1-propanol (PP). These experimental data were fitted to several competitive bi-Langmuir models (with 8, 6, 5 and 4 parameters) and to the competitive Langmuir model. The latter model accounted well for the behavior of both PP enantiomers on Chiracel OB (cellulose tribenzoate coated on silica gel). The parameters obtained were used in numerical calculations to predict the band profiles of the two single components and of their mixtures under overloaded conditions. The equilibrium-dispersive model provides satisfactory results, with minor differences between the calculated and the experimental profiles. These differences became negligible when a more complex kinetic model was used, with a concentration-dependent rate coefficient.

Adsorption↗

Separation of isomeric naphthalenesulphonic acids by micro high-performance liquid chromatography with mobile phases containing cyclodextrin.

Aromatic sulphonic acids are important dye intermediates and the determination of the individual isomers after their preparation by sulphonation of the parent aromatic hydrocarbon is important for the monitoring of the dye production process. For this purpose, either reversed-phase chromatography with mobile phases containing strong electrolytes as additives or capillary zone electrophoresis with working electrolytes containing cyclodextrins can be used to separate and determine not only individual sulphonation products with various numbers of sulphonic groups, but also various isomeric di- and trisulphonic acids. However, the separation of some isomers using either of the two techniques is not fully satisfactory. In the present work, HPLC with mobile phases containing cyclodextrins was employed to improve previously achieved separations of aromatic sulphonic acids. Because of the high cost of cyclodextrin, microcolumn HPLC with diode-array detection on the columns prepared in laboratory by supercritical fluid packing technique was employed for this purpose. Capillary columns packed with various octadecyl silica gel materials were compared and their stability and efficiency were found suitable for the separation of the compounds tested. The selectivity of separation of some isomers improved significantly with respect to the previous methods. Procedures were designed for separation and analytical control of technological processes producing dye intermediates.

Chromatography, High Pressure Liquid↗

Retention mechanism, isocratic and gradient-elution separation and characterization of (co)polymers in normal-phase and reversed-phase high-performance liquid chromatography.

Synthetic (co)polymers or (co)oligomers with two (or more) repeating groups show not only molar mass distribution, but also composition and sequence distribution of the individual repeat units. To characterize such two- (or more-) dimensional distribution, liquid chromatography under "critical conditions" has been suggested, where the separation according to one type of repeating units is suppressed by balancing the adsorption and the size-exclusion effects. In present work it is shown that by combination of adequately selected separation conditions in normal-phase and in reversed-phase systems, the two-dimensional distribution mode can be adjusted to result in the separation following the distribution of any of the two repeat units in ethylene oxide-propylene oxide block (co)oligomers. Based on the retention mechanism suggested, prediction and optimization of the conditions for isocratic and gradient-elution separations of (co)oligomers is possible. HPLC-MS with atmospheric-pressure chemical ionization is a valuable tool for unambiguous identification of the individual (co)oligomers and their tracking in course of method development. Gradient elution can be used for the separation and characterization of block (co)oligomers of ethylene oxide (EO) and propylene oxide (PO) according to the number of the units in one block, while the separation according to the distribution of the units in the other block is suppressed. The effects of the arrangement of the individual EO and PO blocks in the block (co)oligomers (the sequence distribution) affects significantly the retention behavior and the selection of the optimum separation conditions.

Chromatography, High Pressure Liquid↗

Analysis of metal complex azo dyes by high-performance liquid chromatography/electrospray ionization mass spectrometry and multistage mass spectrometry

Five metal complex azo compounds were analyzed using negative-ion electrospray ionization mass spectrometry (ESI-MS). Mass spectra of all compounds yield intense peaks corresponding to [M - H](-) ions without any fragmentation, where M denotes the neutral compound with a proton as the counterion. Under collision induced dissociation (CID) conditions, structurally important fragment ions were studied using the ion trap analyzer with a multistage mass spectrometry (MS(n) facility. Synthesized compounds with (15)N atoms in the azo group facilitated the fragmentation pattern recognition. A reversed-phase high-performance liquid chromatography (HPLC) method using 5 mM ammonium acetate in 70% aqueous acetonitrile as mobile phase was developed making possible the separation of all complex compounds tested. The lower detection limits of the ESI-MS method are in the range 10-20 ng of each compound. The HPLC/ESI-MS method makes possible the monitoring of ligand exchange in aqueous solutions of metal complex azo dyes, and also investigation of the stabilities of the complexes in solution. Copyright 2000 John Wiley & Sons, Ltd.

Journal Article↗

Analytical monitoring of the production of biodiesel by high-performance liquid chromatography with various detection methods.

Gradient elution reversed-phase high-performance liquid chromatography (RP-HPLC) was used for the determination of compounds occurring during the production of biodiesel from rapeseed oil. Individual triacylglycerols (TGs), diacylglycerols, monoacylglycerols and methyl esters of oleic, linoleic and linolenic acids and free fatty acids were separated in 25 min using a combined linear gradient with aqueous-organic and non-aqueous mobile phase steps: 70% acetonitrile+30% water in 0 min, 100% acetonitrile in 10 min, 50% acetonitrile+50% 2-propanol-hexane (5:4, v/v) in 20 min and 5 min final hold-up. Another method with a non-aqueous linear mobile phase gradient [from 100% methanol to 50% methanol+50% 2-propanol-hexane (5:4, v/v) in 15 min] was used for fast monitoring of conversion of rapeseed oil triacylglycerols to fatty acid methyl esters and for quantitation of residual TGs in the final biodiesel product. Sensitivity and linearity of various detection modes (UV detection at 205 nm, evaporative light scattering detection and mass spectrometric detection) were compared. The individual sample compounds were identified using coupled HPLC-atmospheric pressure chemical ionization mass spectrometry in the positive-ion mode.

Chromatography, High Pressure Liquid↗

Interpretation of electrospray and atmospheric pressure chemical ionization mass spectra of 10-formyl-7,8-dihydrofolic acid and 5-formyl-5,6,7,8-tetrahydropteroic acid.

Interpretation of positive- and negative-ion electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) mass spectra of 10-formyl-7,8-dihydrofolic acid (10-FDHFA) and 5-formyl-5, 6,7,8-tetrahydropteroic acid (5-FTHPA) is discussed. ESI mass spectra enable unambiguous molecular weight (MW) determination. In addition to the determination of MW, APCI mass spectra also make possible structure elucidation of 10-FDHFA and 5-FTHPA. ESI and APCI are complementary ionization techniques and appear to be useful alternatives to conventional electron ionization (EI) for the structure elucidation of non-volatile carboxylic acids. Prior to mass spectral analysis, the acids investigated were separated by reverse-phase high-performance liquid chromatography (HPLC).

Atmospheric Pressure↗

Optimization of the recovery yield and of the production rate in overloaded gradient-elution reversed-phase chromatography.

To investigate the effects of various parameters on the production rate and on the recovery yield in overloaded reversed-phase gradient-elution chromatography, band profiles of binary mixtures of phenol and o-cresol were calculated using the experimental parameters of the distribution isotherms determined by binary frontal analysis. The effects of the feed volume and of the steepness of a continuous gradient (gradient time) of methanol in aqueous-organic mobile phases on the separation were studied. If the sample feed in a solvent with weak elution strength is used, combined effects of on-column enrichment, frontal chromatography and sharpening of the later eluted bands may enhance the production rate and the recovery yield. Steep continuous gradients offer better results than isocratic elution if the feed is dissolved in water and injected into the mobile phase with a higher elution strength, because larger volumes of dilute feed can be separated with high recovery yields.

Chromatography, Liquid↗

Fitting competitive adsorption isotherms to the experimental distribution data in reversed-phase systems.

Distribution isotherms of phenol and resorcinol and o-cresol were determined between aqueous solutions with different concentrations of methanol and an octadecyl silica adsorbent. Single-component distribution data can be well described by either the Langmuir or the Jovanovic adsorption isotherms. The competitive Langmuir isotherm with single-component isotherm coefficients can describe moderately successfully the distribution in mixtures of phenol and o-cresol in 20-40% methanol, but it does not fit the experimental distribution data of the mixtures of phenol and resorcinol, which are better described by the competitive Jovanovic and the quadratic isotherms with single-component coefficients. Generally, the quality of the fit of the isotherms improves as the concentration of methanol in the solutions increases. The dependencies of the logarithms of the Langmuir isotherm coefficients and of the coefficients of quadratic and Jovanovic isotherms on the composition of the mobile phase can be described by second-degree polynomial equations over the range 0-40% methanol in water.

Adsorption↗

Description and prediction of retention in normal-phase high-performance liquid chromatography with binary and ternary mobile phase.

The suitabilities of several earlier reported models for description and prediction of retention in normal-phase systems with mobile phases comprised of two organic solvents-a polar and a non-polar one-were tested on the chromatographic behaviour of phenylurea herbicides and alkyl-, aryl- and nitrophenols as sample solutes with a silica-gel column and 2-propanol, n-heptane and dioxane as mobile phase components. With few exceptions, the data obtained from the best-fit three-parameter retention equation differ from the experimental capacity factors at less than 0.1, or 2%, and for most compounds the fit is better than 1%. Of the two two-parameter equations derived from theoretical models, one failed to describe the retention behaviour. The fit of the other two-parameter equation to the retention data of phenols is slightly inferior than with the three-parameter equation. The differences between the fitted data and the experimental capacity factors of substituted phenylureas were < or = 0.2, or 5%. Three-parameter equations were suggested to describe the retention behaviour in three-component mobile phases either at a constant sum or at a constant ratio of the two stronger solvents in ternary mobile phases. To fit the three-component data at any combination of concentrations of the three solvents with the error comparable to that in binary mobile phases, a nine-parameter equation is necessary. Suitability of these equations to describe the experimental behaviour of substituted phenylureas and phenols was verified in ternary mobile phases with various concentrations of 2-propanol, dioxane and n-heptane. Finally, a method was suggested for the prediction of retention in ternary mobile phases with varying concentration ratios of the two polar solvents from the parameters of best-fit equations in binary mobile phases and a single experimental capacity factor in a ternary mobile phase. For most capacity factors, the error of prediction was lower than 0.2, or 5%. Dried solvents were used to improve the reproducibility of the results and the temperature was controlled to +/-0.1 degree C in all experiments. With these precautions, differences between the original retention volumes and the data from repeated experiments measured after ten months of use of the column in the system tested were less than 0.2 ml for 85% of the values compared.

1-Propanol↗