Biomedical subjects

Jean-Luc Veuthey

Publications and source records attributed to Jean-Luc Veuthey.

At least 19 recordsLinked to original sources

PubMed2007

Nonaqueous capillary electrophoresis in pharmaceutical analysis.

This review presents different solvents and electrolytes commonly used as BGEs in NACE for the analysis of pharmaceutical compounds. Most NACE applications carried out since 1998 for the analysis of compounds of pharmaceutical interest are presented in four tables: (i) analysis of drugs and related substances, (ii) analysis of chiral substances, (iii) analysis of phytochemical extracts and (iv) analysis of drugs in biological fluids. These selected examples are used to illustrate the interest in NACE versus conventional aqueous CE.

Chemistry, Pharmaceutical↗

PubMed2006

Recent developments in liquid chromatography--impact on qualitative and quantitative performance.

In order to reduce the analysis time and maintain good efficiency in liquid chromatography (LC), several solutions are currently being investigated. The focus of this study was to compare, both qualitatively and quantitatively, the chromatographic performance of a conventional LC with selected approaches, namely monolithic supports, high temperature LC (up to 90 degrees C), and sub-2 microm particles combined with high pressure (up to 1000 bar). This comparison was achieved from a qualitative point of view with a special attention paid to the analysis of time reduction, efficiency improvement, and pressure constraint. For this purpose, the different approaches were discussed using Knox curves and other kinetic plots. It appeared that columns packed with sub-2 microm particles under high-pressure conditions (UPLC) were well adapted and this option represents an attractive alternative to conventional LC; however, the other alternative approaches should not be neglected. The quantitative evaluation of these techniques was performed on the basis of the validation of results of a pharmaceutical formulation (Rapidocaïne), following SFSTP 2003 guidelines. Fast-LC approaches demonstrated equivalent performance to conventional LC in terms of trueness, precision, and accuracy profile, with a significant time reduction (up to 8x) according to the selected methodology.

Chromatography, Liquid↗

PubMed2006

Evaluation of the influence of protein precipitation prior to on-line SPE-LC-API/MS procedures using multivariate data analysis.

Matrix effects on mass spectrometry (MS) response were investigated with three atmospheric pressure ionization (API) sources after on-line solid-phase extraction (SPE) of human plasma. On-line SPE was evaluated with one restricted access material (RAM), two large particle supports (LPS) and one monolith. A sample protein precipitation (PP) with acetonitrile (2:1) and a direct injection were tested. Principal component analysis (PCA) was performed to simplify data presentation and interpretation. Protein precipitation was found to be mandatory for reducing signal modification. Regarding sensitivity towards matrix effects after PP, atmospheric pressure photoionization (APPI) was globally the least sensitive ionization mode while electrospray ionization ESI was the most sensitive.

Alanine↗

PubMed2006

Analytical discrimination of poisonous and nonpoisonous chemotypes of giant fennel (Ferula communis L.) through their biologically active and volatile fractions.

Giant fennel (Ferula communis L.) from Sardinia is characterized by two chemotypes with different biological activities. One chemotype is poisonous, due to prenylcoumarins, and responsible for ferulosis, which mainly affects sheep and goats, cattle, and horses; the other chemotype is nonpoisonous and contains daucane esters. The two chemotypes cannot be distinguished botanically. High-performance liquid chromatography-diode array-ultraviolet detection-mass spectrometry (HPLC-DAD-UV-MS) analysis of the composition of the fractions containing the biologically active metabolites and of the volatile fractions, by gas chromatography-mass spectrometry (GC-MS), of both essential oil and headspace sampled by headspace solid-phase microextraction (HS-SPME) are here shown to be effective in discriminating the poisonous and nonpoisonous chemotypes. HS-SPME with CAR/PDMS/DVB in combination with GC-MS has also been found to be a successful, fully automated one-step method for rapid and unequivocal discrimination of the two chemotypes, using aristolene and allohedycaryol as markers of the poisonous and nonpoisonous chemotypes, respectively.

Chromatography, High Pressure Liquid↗

PubMed2006

Characterization and comparison of the chromatographic performance of different types of reversed-phase stationary phases.

The chromatographic performance of several base-deactivated stationary phases was evaluated with a specific chromatographic test. Seven basic test compounds, possessing different physico-chemical properties were injected on different supports with two mobile phases: one at pH 7.0 (acetonitrile-phosphate buffer, 40:60, v/v), and the other at pH 3.0 (acetonitrile-phosphate buffer, 15:85, v/v). Chromatographic parameters obtained under these conditions were treated by principal component analysis (PCA) to separate base deactivated supports according to their silanol activity (pH 7.0 mobile phase) and hydrophobic properties (pH 3.0 mobile phase). The information given by the specific test column evaluation was improved with complementary chemometric tools such as hierarchical cluster analysis. The same base deactivated supports were also tested following a general test procedure issued from the literature and obtained fundamental properties (in particular silanol activity and hydrophobicity) were compared with column evaluation obtained with the specific test: results were in good agreement, although the use of the specific test offered a better differentiation between numerous base-deactivated supports.

Chemical Phenomena↗

PubMed2006

Chromatographic behaviour and comparison of column packed with sub-2 microm stationary phases in liquid chromatography.

In order to reduce the analysis time and maintain good efficiency in liquid chromatography, it is advisable to simultaneously decrease the column length and the particle size of the chromatographic support. Therefore, several manufacturers have developed and commercialized short columns filled with particles that have a diameter smaller than 2 microm. The focus of this work was to check the chromatographic performance of such columns and compare possibilities offered by sub-2 microm supports with conventional columns in terms of analysis time reduction and efficiency improvements. For this purpose, different parameters were discussed namely: separation impedance (E), Knox curves (h,v), and number of plates by time unit (N/t0). Kinetic plots were also drawn. It appeared that sub-2 microm supports were well adapted to improve chromatographic performance and to reduce the analysis time. Furthermore, it was also demonstrated that the best chromatographic performances were reached with high pressure systems (up to 1000 bar).

Chromatography, Liquid↗

PubMed2006

Practical constraints in the kinetic plot representation of chromatographic performance data: theory and application to experimental data.

It is demonstrated that the kinetic plot representation of experimental plate height data can also account for practical constraints on the column length, the peak width, the viscous heating, and the mobile-phase velocity without needing any iterative solution routine. This implies that the best possible kinetic performance to be expected from a given tested support under any possible set of practical optimization constraints can always be found using a directly responding calculation spreadsheet template. To show how the resulting constrained kinetic plots can be used as a powerful design and selection tool, the method has been applied to a series of plate height measurements performed on a number of different commercial columns for the same component (butyl-parabene) and mobile-phase composition. The method, for example, allows one to account for the fact that the advantageous solutions displayed by the silica monolith and 5 microm particle columns in the large plate number range of the free kinetic plot are no longer accessible if applying a maximal column length constraint of Lmax = 30 cm. In the plate number range that remains accessible, the investigated sub-2 mum particle columns in any case perform (at least for the presently considered parabene separation) better than the 3.5 mum particle columns or silica monolith, especially if considering the use of system pressures exceeding 400 bar. The constrained kinetic plot method can also be used to select the best-suited column length from an available product gamma to perform a separation with a preset number of plates. One of the optimization results that is obtained in this case is that sometimes a significant gain in analysis time can be obtained by selecting a longer column, yielding the desired plate number at a larger velocity than that for a shorter column.

Algorithms↗

PubMed2006

Enhanced method performances for conventional and chiral CE-ESI/MS analyses in plasma.

Due to its high efficiency, selectivity, and sensitivity, CE-ESI/MS has evolved as an efficient technique for the drugs and metabolites analysis in biological matrices. However, a sample preparation is mandatory prior to CE-ESI/MS analysis. To achieve fast and simplified sample preparation of plasma samples, protein precipitation (PP) and liquid-liquid extraction (LLE) were used with two injection techniques: hydrodynamic (HD) and electrokinetic (EK) injection. CE-ESI/MS analyses of pharmaceutical compounds and amphetamine derivatives were developed. Detection limits of 1 ppm were reached with PP and HD injection whereas 1 ppb was detected when samples were prepared with LLE and injected by EK. Same experiments were performed for stereoselective determinations in partial-filling mode and detection limits achieved were equivalent to conventional analysis (0.5 ppb per enantiomer). When complex matrices are analyzed, MS signal suppression or enhancement effects are generally not reproducible and could compromise results with ESI. Therefore, matrix effect was investigated in CE-ESI/MS with a commercially available coaxial sheath-liquid ESI interface used as postcapillary infusion system to determine MS signal alterations. Matrix effects were differentially evidenced according to the selected sample preparation. With PP, signal suppression was observed out of the analyses migration window, while for LLE no relevant matrix effect occurred in all experiments.

Amphetamines↗

PubMed2006

Interlaboratory study of a NACE method for the determination of R-timolol content in S-timolol maleate: assessment of uncertainty.

Analyses of statistical variance were applied to evaluate the precision and practicality of a CD-based NACE assay for R-timolol after enantiomeric separation of R- and S-timolol. Data were collected in an interlaboratory study by 11 participating laboratories located in Europe and North America. General qualitative method performance was examined using suitability descriptors (i.e. resolution, selectivity, migration times and S/N), while precision was determined by quantification of variances in the determination of R-timolol at four different impurity levels in S-timolol maleate samples. The interlaboratory trials were designed in accordance with the ISO guideline 5725-2. This allowed estimating for each sample, the different variances, i.e. between-laboratory (s2(Laboratories)), between-day (s2(Days)) and between-replicate (s2(Replicates)). The variances of repeatability (s2r) and reproducibility (s2R) were then calculated. The estimated uncertainty, derived from the precision estimates, seems to be concentration-dependent above a given threshold. This example of R-timolol illustrates how a laboratory can evaluate uncertainty in general.

Adrenergic beta-Antagonists↗

PubMed2006

Analysis of isomeric tropane alkaloids from Schizanthus grahamii by very fast gas chromatography.

This study presents a very fast GC analysis applied for the baseline separation of isomeric tropane alkaloids extracted from the stem-bark of Schizanthus grahamii (Solanaceae). The work provided a challenging application where isothermal analysis in conjunction with very short narrow bore columns (3 m x 100 microm ID and 1.5 m x 50 microm ID) was particularly suited for the speeding up. Experimental parameters were used in the optimisation steps, including selection of stationary phase, temperature, internal column diameter and optimal practicable gas velocity. Some considerations about sample injection in fast isothermal analysis are also briefly presented. Finally, the investigated approach allowed a very fast baseline separation of four positional and configurational isomers in less than 9 s.

Chromatography, Gas↗

PubMed2006

Fast analysis in liquid chromatography using small particle size and high pressure.

In order to enhance chromatographic performances in terms of efficiency and rapidity, LC has recently evolved in the development of short columns packed with small particles (sub-2 microm) working at high pressures (> 400 bar). This approach has been described 30 years ago according to the fundamental chromatographic equations. However, systems and columns compatible with such high pressures have been introduced in the market in 2004 only. Advantages of small particles working at high pressure will be discussed in terms of sensitivity, efficiency, resolution, and analysis time. Potential problems encountered with high pressure in terms of frictional heating and solvent compressibility will also be discussed even if systems working at a maximum pressure of 1000 bar are not influenced by these parameters and give reliable and reproducible results. Several applications will highlight the potential and interest of this new technology.

Chromatography, Liquid↗

PubMed2006

Evaluation of solid-phase microextraction desorption parameters for fast GC analysis of cocaine in coca leaves.

By its simplicity and rapidity, solid-phase microextraction (SPME) appears as an interesting alternative for sample introduction in fast gas chromatography (fast GC). This combination depends on numerous parameters affecting the desorption step (i.e., the release of compounds from the SPME fiber coating to the GC column). In this study, different liner diameters, injection temperatures, and gas flow rates are evaluated to accelerate the thermal desorption process in the injection port. This process is followed with real-time direct coupling a split/splitless injector to a mass spectrometer by means of a short capillary. It is shown that an effective, quantitative, and rapid transfer of cocaine (COC) and cocaethylene (CE) is performed with a 0.75-mm i.d. liner, at 280 degrees C and 4 mL/min gas flow rate. The 7-microm polydimethylsiloxane (PDMS) coating is selected for combination with fast GC because the 100-microm PDMS fiber presents some limitations caused by fiber bleeding. Finally, the developed SPME-fast GC method is applied to perform in less than 5 min, the quantitation of COC extracted from coca leaves by focused microwave-assisted extraction. An amount of 7.6 +/- 0.5 mg of COC per gram of dry mass is found, which is in good agreement with previously published results.

Chromatography, Gas↗

PubMed2005

Focused microwave-assisted extraction combined with solid-phase microextraction and gas chromatography-mass spectrometry for the selective analysis of cocaine from coca leaves.

An effective combination of focused microwave-assisted extraction (FMAE) with solid-phase microextraction (SPME) prior to gas chromatography (GC) is described for the selective extraction and quantitative analysis of cocaine from coca leaves (Erythroxylum coca). This approach required switching from an organic extraction solvent to an aqueous medium more compatible with SPME liquid sampling. SPME was performed in the direct immersion mode with a universal 100 microm polydimethylsiloxane (PDMS) coated fibre. Parameters influencing this extraction step, such as solution pH, sampling time and temperature are discussed. Furthermore, the overall extraction process takes into account the stability of cocaine in alkaline aqueous solutions at different temperatures. Cocaine degradation rate was determined by capillary electrophoresis using the short end injection procedure. In the selected extraction conditions, less than 5% of cocaine was degraded after 60 min. From a qualitative point of view, a significant gain in selectivity was obtained with the incorporation of SPME in the extraction procedure. As a consequence of SPME clean-up, shorter columns could be used and analysis time was reduced to 6 min compared to 35 min with conventional GC. Quantitative results led to a cocaine content of 0.70 +/- 0.04% in dry leaves (RSD <5%) which agreed with previous investigations.

Chemical Fractionation↗

PubMed2005

Cocaine distribution in wild Erythroxylum species.

Cocaine distribution was studied in leaves of wild Erythroxylum species originating from Bolivia, Brazil, Ecuador, Paraguay, Peru, Mexico, USA, Venezuela and Mauritius. Among 51 species, 28 had never been phytochemically investigated before. Cocaine was efficiently and rapidly extracted with methanol, using focused microwaves at atmospheric pressure, and analysed without any further purification by capillary gas chromatography coupled to mass spectrometry. Cocaine was reported for the first time in 14 species. Erythroxylum laetevirens was the wild species with the highest cocaine content. Its qualitative chromatographic profile also revealed other characteristic tropane alkaloids. Finally, its cocaine content was compared to those of two cultivated coca plants as well as with a coca tea bag sample.

Coca↗

PubMed2005

Strategies for rapid chiral analysis by capillary electrophoresis.

The aim of this study was to investigate four strategies to decrease chiral CE analysis time: (1) short-end injection technique, (2) high electric field through a capillary length reduction, (3) external pressure application and (4) capillary dynamically coated to generate an important electroosmotic flow. These approaches were applied for a simultaneous enantiomeric separation of amphetamine and four related compounds using a neutral derivatised cyclodextrin (hydroxypropyl-beta-cyclodextrin) as chiral selector. Analysis time and CE performances, in terms of peak efficiency and resolution, were examined. Among the investigated strategies, the dynamic coating procedure appeared to be the most suitable approach to decrease analysis time (inferior to 7 min) and improve sensitivity. Furthermore, it exhibited very good migration time repeatability (0.1%). This benefit is of utmost interest in chiral analysis for an unambiguous peak identification, especially for a complex mixture such as reported in this study.

Amphetamines↗

PubMed2005

A study of interlaboratory influence on column evaluation.

A liquid chromatography method for the characterization of base deactivated columns was investigated in a collaborative study involving six laboratories. This work was carried out on two chromatographic supports (Xterra RP 18 and Symmetry Shield). Different cooling systems, namely water bath and air oven, were tested and it was shown that column thermoregulation did not significantly influence chromatographic data. In order to control the mobile phase composition, the latter was prepared by weight rather than volume. Thanks to the injection of a set of selected neutral compounds, extra-column effects were evaluated in each of the participating laboratories. The results showed that chromatographic supports tested in different laboratories and following the same test protocol could be effectively compared.

Chromatography, High Pressure Liquid↗

PubMed2005

Use of porous graphitic carbon column for the separation of natural isomeric tropane alkaloids by capillary LC and mass spectrometry.

This study presents the outstanding chromatographic selectivity of a porous graphitic carbon support for the separation of four isomeric tropane alkaloids from the stem-bark of Schizanthus grahamii (Solanaceae). Capillary liquid chromatography coupled to mass spectrometry was studied after the appropriate selection of mobile phase composition, temperature, nature and concentration of the acidic modifier. Fragmentation behaviour by in-source collision-induced dissociation (CID) on a single quadrupole mass spectrometer, or MSn with an ion-trap, was investigated for structural identification. The ability to differentiate the isomers by in-source CID was demonstrated and a fragmentation pathway, based on MSn sequences together with accurate mass experiments, was proposed.

Alkaloids↗

PubMed2005

Determination of electroosmotic flow in nonaqueous capillary electrophoresis.

Mobility of the electroosmotic flow (mu(EOF)) in fused-silica capillaries strongly depends on the nature of the background electrolyte. In this study, 27 solvent systems were investigated, namely water, methanol, ethanol, 2-propanol, 1-butanol, acetonitrile (MeCN), formamide, N-methylformamide (NMF), N,N-dimethylformamide and dimethyl sulfoxyde, as well as 8 hydroorganic and 9 organic mixtures. For each system, six mu(EOF) were determined at a different ionic strength in basic conditions, and an absolute electroosmotic flow mobility (mu(EOF,0)) was extrapolated according to the Debye-Huckel Onsager model. The obtained mu(EOF,0) values were correlated with the solvent's relative permittivity (epsilon) and viscosity (eta). A good correlation (r2=0.867) between mu(EOF,0) and the solvent's epsilon/eta ratio was demonstrated, except for two solvents (MeCN and NMF). Furthermore, the donor number (DN) of a solvent took into account the possible zeta potential modification in the electric double layer near the capillary wall. Consequently, the relationship between mu(EOF,0) and epsilon/(eta x DN) was superior, with a r2 of 0.943 for 10 pure solvents.

Electrophoresis, Capillary↗