A multianalyte ELISA for immunochemical screening of sulfonamide,fluoroquinolone and ß-lactam antibiotics in milk samples using class-selective bioreceptors

A multianalyte ELISA has been developed for the simultaneous determination of the most frequently used antibiotic families in the veterinary field following the typical planar microarray configuration, where the identity of the target analyte is encoded by its location in the detection platform (Master et al. in Drug Discovery Today 11:1007-1011, 2006). To accomplish this aim, two individual enzyme-linked immunosorbent assays for sulfonamide and fluoroquinolone antibiotics and an enzyme-linked receptor assay for ss-lactam antibiotics have been combined. The strategy uses microplates coated with the corresponding haptenized proteins in specific sections of the microplate. The samples are mixed with a cocktail containing the bioreagents, and distributed in the wells of the microplate. Identification of the antibiotic present in a particular sample is consequently accomplished by detecting a positive response on the corresponding microplate section. Since the bioreceptors used show a wide recognition of the congeners of each antibiotic family, the multianalyte method is able to detect more than 25 different antibiotics from the three most important antibiotic families. The detectability reached in full-fat milk samples is below the European maximum residue limits. The accuracy and reliability of this multiplexed bioanalytical method have been demonstrated by analyzing blind spiked samples.     

Preparation of second generation ionic liquids by efficient solvent-free alkylation of N-heterocycles with chloroalkanes

Non-conventional techniques, such as microwave (MW) and power ultrasound (US) as well as combined MW/US irradiation, have been used to promote one-pot synthesis of second-generation ionic liquids (ILs), cutting down reaction times and improving yields. However, the use of chloroalkanes in the alkylation of N-heterocycles requires more drastic conditions if results are to match those obtained with more reactive alkyl halides. The present paper describes a series of MW- or MW/US-promoted IL preparations starting from chloroalkanes and classic heterocycles (1-methylimidazole, pyridine and 1-methylpyrrolidine). When reactions were carried out under conventional heating in an oil bath they required longer reaction times and gave poorer yields. (1)H-NMR analysis and ion-exchange chromatography showed that the present solventless procedure afforded ILs of satisfactory purity. The observed high yields (usually 70-98% isolated), and short reaction times showed that a straightforward access to ILs can be also achieved with the use of alkyl chlorides, resulting in a considerable reduction of costs.     

Incoherent quasielastic neutron scattering study of molecular dynamics of 4-n-octyl-4'-cyanobiphenyl

We report incoherent quasielastic neutron scattering experiments on the thermotropic liquid crystal 4-n-octyl-4'-cyanobiphenyl. The combination of time-of-flight and backscattering data allows analysis of the intermediate scattering function over about three decades of relaxation times. Translational diffusion and uniaxial molecular rotations are clearly identified as the major relaxation processes in, respectively, the nanosecond and picosecond time scales.     

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

Electrokinetic supercharging has been integrated in CZE for the development of a highly sensitive methodology for protein tryptic digest analysis. A careful choice of the experimental conditions led to sensitivity enhancement factors between 1000 and 10,000 whilst maintaining a satisfactory resolution. Peptides in the low nanomolar concentration range have been detected despite the use of the poorly sensitive UV absorbance detection mode. The buffer system used in this study is fully suitable for coupling CE to MS.     

Inner membrane lipids of Escherichia coli form domains

In prokaryotic cells, the hypothesis of the existence of lipid domains was considered. In order to test this hypothesis and study the organization of lipids in the inner membrane of Escherichia coli, we elaborated Langmuir films mimicking the inner leaflet of this membrane by considering lipids extracted from the inner membrane of E coli by Folch protocol.

Lipid monolayers were elaborated by using these extracts (Langmuir technique); the organization of the resulting films was studied at the air–water interface by Brewster angle microscopy and after transfer onto muscovite by atomic force microscopy. The existence of domains was demonstrated for different interfacial pressures of biological interest, and their stability was studied.     

Extraction and purification procedures for simultaneous quantification of phenolic xenoestrogens and steroid estrogens in river sediment by gas chromatography/ion trap mass spectrometry

A sensitive and simple method based on ultrasonication extraction with a hexane/acetone (2:1, v/v) mixture, followed by clean up of the extract by solid-phase extraction (SPE) and gas chromatography/mass spectrometry (GC/MS) detection, has been developed and validated for the analysis of 20 estrogenic endocrine-disrupting chemicals (EEDCs) including phenolic xenoestrogens, synthetic and natural estrogens in river sediment. After extraction and purification, analytes are derivatised with a BSTFA/TMCS/pyridine (49:1:50, v/v/v) mixture and quantified by GC/MS. The GC/MS method involves switching between electron ionisation (EI) and chemical ionisation (CI); it also switches between selected ion storage and tandem mass spectrometry detection. The applicability of the method has been demonstrated by analysing extracts of French river sediments for which bioanalytical tests (in vitro) had already shown that they were impacted by estrogenic endocrine disrupters. The biological contribution of all the products detected in each sediment extract was compared to the estrogenic activity measured by bioassays.     

Elucidation of nitrate reduction pathways in anaerobic bioreactors using a stable isotope approach

Leachate recirculation allows an increase of moisture content and the enhancement of the anaerobic digestion of wastes in landfill. Since there is no ammonia elimination process in landfill when leachate is recirculated, NH(4) (+) may accumulate. One strategy for NH(4) (+) removal is to treat aerobically the leachate outside the landfill to convert NH(4) (+) into NO(3) (-). When nitrified leachate is recirculated, denitrification should occur in the waste. We have previously shown that wastes have a large capacity to convert nitrate into N(2). Nevertheless, in some cases we observed nitrate reduction without gaseous nitrogen production. Using a stepwise multiple regression models, H(2)S concentration was the unique parameter found to have a negative effect on N(2) production. We then suspected that dissimilatory nitrate reduction to ammonium (DNRA) occurred in the presence of H(2)S. In order to verify this hypothesis, (15)N nitrate injections were performed into microcosms containing different H(2)S concentrations. The ammonium (15)N enrichment was measured using an elemental analyser coupled to an isotope ratio mass spectrometer. In the two microcosms containing the highest H(2)S concentrations, the ammonium was (15)N enriched and at the end of the experiment all the added nitrate was converted into ammonium. For the two microcosms containing the lowest H(2)S concentrations, no (15)N enrichment of ammonium was observed. This isotopic approach has allowed us to demonstrate that, in the presence of significant concentrations of H(2)S, denitrification is replaced by DNRA.