Le Centre de Compétence en Chimie et Toxicologie Analytiques. The Centre of Competence in Analytic Chemistry and Toxicology (CCCTA)

This article presents concisely activities, goals and members of the Centre of Competence in Analytical Chemistry and Toxicology which was created recently in the Lake Geneva region

What Type of Research Is Done in the Laboratory of Pharmaceutical Analytical Chemistry of the University of Geneva?

This article presents briefly the main research projects developed in the Laboratory of Pharmaceutical Analytical Chemistry at the University of Geneva. This laboratory is particularly concerned about enantiomeric separation of drugs and phytochemical analysis. In this context, techniques such as gas, liquid and supercritical fluid chromatography as well as capillary electrophoresis are studied. Furthermore, the development of new sample preparation techniques is investigated

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°C and 4 mL/min gas flow rate. The 7-µm polydimethylsiloxane (PDMS) coating is selected for combination with fast GC because the 100- µm 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 result

Comparative study of recent wide-pore materials of different stationary phase morphology, applied for the reversed-phase analysis of recombinant monoclonal antibodies

Various recent wide-pore reversed-phase stationary phases were studied for the analysis of intact monoclonal antibodies (mAbs) of 150kDa and their fragments possessing sizes between 25 and 50kDa. Different types of column technology were evaluated, namely, a prototype silica-based inorganic monolith containing mesopores of ∼250Å and macropores of ∼ 1.1μm, a column packed with 3.6μm wide-pore core-shell particles possessing a wide pore size distribution with an average around 200Å and a column packed with fully porous 1.7μm particles having pore size of ∼300Å. The performance of these wide-pore materials was compared with that of a poly(styrene-divinyl benzene) organic monolithic column, with a macropore size of approximately 1μm but without mesopores (stagnant pores). A systematic investigation was carried out using model IgG1 and IgG2 mAbs, namely rituximab, panitumumab, and bevacizumab. Firstly, the recoveries of intact and reduced mAbs were compared on the two monolithic phases, and it appeared that adsorption was less pronounced on the organic monolith, probably due to the difference in chemistry (C18 versus phenyl) and the absence of mesopores (stagnant zones). Secondly, the kinetic performance was investigated in gradient elution mode for all columns. For this purpose, peak capacities per meter as well as peak capacities per time unit and per pressure unit (PPT) were calculated at various flow rates, to compare performance of columns with different dimensions. In terms of peak capacity per meter, the core-shell 3.6μm and fully porous 1.7μm columns outperformed the two monolithic phases, at a temperature of 60°C. However, when considering the PPT values, the core-shell 3.6μm column remained the best phase while the prototype silica-based monoliths became very interesting, mostly due to a very high permeability compared with the organic monolith. Therefore, these core-shell and silica-based monolith provided the fastest achievable separation. Finally, at the maximal working temperature of each column, the core-shell 3.6μm column was far better than the other one, because it is the only one stable up to 90°C. Lastly, the loading capacity was also measured on these four different phases. It appeared that the organic monolith was the less interesting and rapidly overloaded, due to the absence of mesopores. On the other hand, the loading capacity of prototype silica-based monolith was indeed reasonabl

New trends in fast and high-resolution liquid chromatography: a critical comparison of existing approaches

Recent developments in chromatographic supports and instrumentation for liquid chromatography (LC) are enabling rapid and highly efficient separations. Various analytical strategies have been proposed, for example the use of silica-based monolithic supports, elevated mobile phase temperatures, and columns packed with sub-3μm superficially porous particles (fused core) or with sub-2μm porous particles for use in ultra-high-pressure LC (UHPLC). The purpose of this review is to describe and compare these approaches in terms of throughput and resolving power, using kinetic data gathered for compounds with molecular weights ranging between 200 and 1300g mol−1 in isocratic and gradient modes. This study demonstrates that the best analytical strategy should be selected on the basis of the analytical problem (e.g., isocratic vs. gradient, throughput vs. efficiency) and the properties of the analyte. UHPLC and fused-core technologies are quite promising for small-molecular-weight compounds, but increasing the mobile phase temperature is useful for larger molecules, for example peptides. Figure Recent progress in HPLC technology to increase throughput and resolving powe

High-Throughput Screening of Drugs of Abuse in Urine by Supported Liquid-Liquid Extraction and UHPLC Coupled to Tandem MS

A qualitative method, involving supported liquid-liquid extraction (SLE) and ultra high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS-MS), was developed for the rapid tentative identification of various drugs of abuse in urine. In this study, 28 drugs and metabolites were covered by the screening procedure. Before analysis, urine samples were extracted by SLE and good extraction recoveries were obtained for most investigated compounds. The UHPLC strategy was then selected for the rapid separation of amphetamines, cocaine, opiates and related compounds in urine. Using columns packed with sub-2µm particles, analysis time was reduced down to 2min, while maintaining acceptable performance. Finally, the detection was by tandem MS operating in the single reaction monitoring (SRM) mode. The most intense transition was selected for the different drugs and SRM dwell times set at 5ms, to maintain sufficient data points across the narrow UHPLC peaks. The tentative identification of the drugs of interest, including amphetamines, opiates and cocaine, was based on both, retention times and mass spectrometry information. With the proposed method, limits of detection were estimated at about 1ngmL−1 and the applicability was assessed by successfully analyzing several samples of drug abusers. Finally, this study demonstrates the potential of UHPLC coupled to tandem MS for the rapid screening of drugs of abuse in urin

Quantitative Capillary Supercritical Fluid Chromatography and Supercritical Fluid Extraction of Basic Drugs of Abuse

Capillary SFC of basic drugs of abuse have been studied using two different columns, and the results were compared with GC. These results have proved the ability of supercritical CO2 to solubilize and extract basic drugs of interest. A novel restrictor has been used to control precisely the flow rate through the extraction cell over a long period of time. Optimal conditions for quantitative recoveries of aqueous morphine adsorbated on C18 silica by SFE has been determined. Recoveries of 100% with a variation of 2% was obtained with an extraction time of 25 min. The influence of the pressure, solvent modifier, and flow rate of extraction fluid on the extraction efficiency was determined. These conditions were also found to be applicable for extracting other drugs of abuse

Quantitative Capillary Supercritical Fluid Chromatography and Supercritical Fluid Extraction of Basic Drugs of Abuse

Capillary SFC of basic drugs of abuse have been studied using two different columns, and the results were compared with GC. These results have proved the ability of supercritical CO2 to solubilize and extract basic drugs of interest. A novel restrictor has been used to control precisely the flow rate through the extraction cell over a long period of time. Optimal conditions for quantitative recoveries of aqueous morphine adsorbated on C18 silica by SFE has been determined. Recoveries of 100% with a variation of 2% was obtained with an extraction time of 25 min. The influence of the pressure, solvent modifier, and flow rate of extraction fluid on the extraction efficiency was determined. These conditions were also found to be applicable for extracting other drugs of abuse

Motifs tree: a new method for predicting post-translational modifications

Motivation: Post-translational modifications (PTMs) are important steps in the maturation of proteins. Several models exist to predict specific PTMs, from manually detected patterns to machine learning methods. On one hand, the manual detection of patterns does not provide the most efficient classifiers and requires an important workload, and on the other hand, models built by machine learning methods are hard to interpret and do not increase biological knowledge. Therefore, we developed a novel method based on patterns discovery and decision trees to predict PTMs. The proposed algorithm builds a decision tree, by coupling the C4.5 algorithm with genetic algorithms, producing high-performance white box classifiers. Our method was tested on the initiator methionine cleavage (IMC) and Nα-terminal acetylation (N-Ac), two of the most common PTMs. Results: The resulting classifiers perform well when compared with existing models. On a set of eukaryotic proteins, they display a cross-validated Matthews correlation coefficient of 0.83 (IMC) and 0.65 (N-Ac). When used to predict potential substrates of N-terminal acetyltransferaseB and N-terminal acetyltransferaseC, our classifiers display better performance than the state of the art. Moreover, we present an analysis of the model predicting IMC for Homo sapiens proteins and demonstrate that we are able to extract experimentally known facts without prior knowledge. Those results validate the fact that our method produces white box models. Availability and implementation: Predictors for IMC and N-Ac and all datasets are freely available at http://terminus.unige.ch/. Contact: [email protected] Supplementary information: Supplementary data are available at Bioinformatics onlin