Multimodal open-tubular capillary electrochromatographic analysis of amines and peptides

This study describes a comparison of different modes of open-tubular electrochromatography (OTCEC) in bare and etched capillaries. To carry out the investigation, the separation of impurities of two synthetic peptides and the separation of a mixture of five heterocyclic aromatic amines were studied. Three different types of stationary phase were evaluated: (i) fluorosurfactants (anionic and zwitterionic) adsorbed in the inner wall of the capillary (electrochromatography with dynamically modified stationary phases (DMS)CEC); (ii) physically adsorbed polymers (DMA-SO(3-) and DMA-N(+)(CH(3))(3)) and (iii) chemically modified capillaries (C(18), cholesteryl 10-undecanoate and diol). The results confirm that electrochromatography can be a viable alternative to capillary electrophoresis (CE) and liquid chromatography, more established separation techniques. It is possible to differentiate some minor species for the synthetic peptides that cannot be resolved by CE or high-performance liquid chromatography (HPLC). Moreover the separation of the amine mixture depends strongly on the stationary phase used.     

Trace determination of cannabinoids and opiates in wastewater and surface waters by ultra-performance liquid chromatography-tandem mass spectrometry

A fast and reliable method using solid-phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) has been developed for the simultaneous detection, identification and quantification of several central nervous system depressor drugs of abuse such as cannabinoids (Delta9-tetrahydrocannabinol, THC) and opiates (morphine, codeine, heroin, methadone, fentanyl) and their metabolites in water samples. Compounds were extracted from water by using Oasis HLB cartridges. After SPE enrichment, the selected depressor drugs, under UPLC optimized conditions, were separated in less than 8 min. Electrospray (ESI) tandem MS in positive ion mode and selected reaction monitoring was used for quantification. ESI-MS/MS conditions such as capillary and cone voltages, source and desolvation temperatures and cone and desolvation gas flow rates have been optimized and MS and MS/MS spectra of the studied compounds were obtained. At the working conditions four identification points were obtained as required by European Union guidelines for analysis by LC-MS/MS. Quality parameters (intra-day and inter-day precisions) for each analyte have been established in three different matrixes (purified, surface and waste waters). Recoveries were generally higher than 70% and instrumental quantification limits and limits of quantification were in the low pg and ng/l range, respectively. Finally, the method has been applied to the analysis of influent and effluents wastewaters and natural water samples from Catalonia (NE Spain) where the presence of several opiates such as morphine, codeine, norcodeine 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine and methadone and cannnabinoids such as THC and 11-nor-carboxy-Delta9-tetrahydrocannabinol has been demonstrated.     

Gas chromatography/ion trap tandem mass spectrometry for the analysis of halobenzenes in soils by solid-phase microextraction

Headspace solid-phase microextraction combined with gas chromatography/ion trap tandem mass spectrometry (HS-SPME/GC/ITMS/MS) was used for the analysis of 12 halobenzenes from soil samples. For MS/MS optimisation, the experiments were performed by precursor ion selection and software controlled operations. Collision-induced dissociation (CID) can be achieved by two different approaches, resonant and non-resonant excitation modes. Different results were obtained using the two approaches, and the resonant excitation mode was chosen as the best for all halobenzenes. Parameters such as the CID excitation amplitude, excitation RF storage level and CID bandwidth frequency were optimised to maximise the formation of halobenzene product ions. A 100-microm polydimethylsiloxane fibre was used for the isolation and preconcentration of the analytes. The HS-SPME/GC/ITMS/MS method was applied to the analysis of halobenzenes in an agricultural soil sample. The halobenzenes were quantified by standard addition, which led to good reproducibility (RSD between 4.7 and 9.2%) and detection limits in the low pg/g range. The method was validated by comparing the results with those obtained in a European inter-laboratory exercise.     

Simultaneous pressurized liquid extraction and clean-up for the analysis of polybrominated biphenyls by gas chromatography-tandem mass spectrometry

This paper describes a fast and simple pressurized liquid extraction (PLE) method combined with gas chromatography coupled to ion trap tandem mass spectrometry (GC-ITMS-MS) for the determination of polybrominated biphenyls (PBBs) in fish samples. The method is based on a simultaneous extraction/clean-up step to reduce analysis time and solvent consumption. The effect of several PLE operating conditions, such as solvent type, extraction temperature and time, number of cycles, and lipid retainer, was optimized to obtain maximum recovery of the analytes with the minimum presence of matrix-interfering compounds. The best conditions were obtained at 100 °C with n-hexane using 15 g of silica modified with sulphuric acid (44%, w/w) as sorbent for lipid removal. Quality parameters of the GC-ITMS-MS method were established, achieving good linearity (r > 0.998), between 1 and 500 ng ml⁻¹, and low instrumental limits of detection (0.14-0.76 pg injected). For the whole method, limits of detection ranging from 0.03 to 0.16 ng g⁻¹ wet weight and good precision (RSD < 16%) were obtained.     

AGNES at vibrated gold microwire electrode for the direct quantification of free copper concentrations

The free metal ion concentration and the dynamic features of the metal species are recognized as key to predict metal bioavailability and toxicity to aquatic organisms. Quantification of the former is, however, still challenging. In this paper, it is shown for the first time that the concentration of free copper (Cu²⁺) can be quantified by applying AGNES (Absence of Gradients and Nernstian equilibrium stripping) at a solid gold electrode. It was found that: i) the amount of deposited Cu follows a Nernstian relationship with the applied deposition potential, and ii) the stripping signal is linearly related with the free metal ion concentration. The performance of AGNES at the vibrating gold microwire electrode (VGME) was assessed for two labile systems: Cu-malonic acid and Cu-iminodiacetic acid at ionic strength 0.01 M and a range of pH values from 4.0 to 6.0. The free Cu concentrations and conditional stability constants obtained by AGNES were in good agreement with stripping scanned voltammetry and thermodynamic theoretical predictions obtained by Visual MinteQ. This work highlights the suitability of gold electrodes for the quantification of free metal ion concentrations by AGNES. It also strongly suggests that other solid electrodes may be well appropriate for such task. This new application of AGNES is a first step towards a range of applications for a number of metals in speciation, toxicological and environmental studies for the direct determination of the key parameter that is the free metal ion concentration.     

Analysis of polychlorinated n-alkanes in environmental samples

Polychlorinated n-alkanes (PCAs), also known as chlorinated paraffins (CPs), are highly complex technical mixtures that contain a huge number of structural isomers, theoretically more than 10,000 diastereomers and enantiomers. As a consequence of their persistence, tendency to bioaccumulation, and widespread and unrestricted use, PCAs have been found in aquatic and terrestrial food webs, even in rural and remote areas. Recently, these compounds have been included in regulatory programs of several international organizations, including the US Environmental Protection Agency and the European Union. Consequently, there is a growing demand for reliable methods with which to analyze PCAs in environmental samples. Here, we review current trends and recent developments in the analysis of PCAs in environmental samples such as air, water, sediment, and biota. Practical aspects of sample preparation, chromatographic separation, and detection are covered, with special emphasis placed on analysis of PCAs using gas chromatography–mass spectrometry. The advantages and limitations of these techniques as well as recent improvements in quantification procedures are discussed.     

Determination of ebrotidine and its metabolites by capillary electrophoresis with UV and mass spectrometry detection

This study describes the application of capillary electrophoresis (CE) to the analysis of ebrotidine and its metabolites as an alternative analytical technique to liquid chromatography. Comparison between UV-diode array spectroscopy and mass spectrometry (MS) using an ion-trap system with electrospray ionization as detection systems has been performed. The quality parameters of the UV detection method were established, obtaining linear calibration curves over the range studied (8-200 mg ml(-1)), limits of detection between 3.4 and 4.3 microg ml(-1), and run-to-run and day-to-day precision lower than 14%. For these compounds the protonated species [M+H]+ and, in some cases, sodium adducts were observed in the MS spectra. Using MS coupled to CE, limits of detection were between 0.5 and 2.6 microg ml(-1).     

Multivariate calibration methods for quantification in strongly overlapping capillary electrophoretic peaks

Capillary zone electrophoresis with diode-array detection was applied to the separation of ebrotidine and its metabolites. However, three of these, which are neutral in the conditions studied, co-migrated with the electroosmotic flow signal. Therefore, strongly overlapping peaks were observed. The main aim of this study was to show the potentiality of capillary electrophoresis in combination with chemometrics. Multivariate calibration methods were applied to quantify these analytes in synthetic mixtures. The results obtained using partial least squares (PLS) were in agreement with actual values, with an overall prediction error of 9.7%.     

Analysis of glycosaminoglycan monosaccharides by capillary electrophoresis using indirect laser-induced fluorescence detection

Two methods for monosaccharide analysis by capillary electrophoresis (CE) using counterelectroosmotic and coelectroosmotic modes with indirect laser-induced fluorescence detection were optimised and compared. A mixture of seven glycosaminoglycan-derived hexoses was separated in alkaline fluorescein-based electrolytes and detected in both counterelectroosmotic and coelectroosmotic conditions. The fluorescein concentration and pH of the background electrolyte, and the influence of the reversal of electroosmotic flow by addition of hexadimethrine bromide on the separation were studied. Coelectroosmotic CE conditions provided better resolution and limits of detection. A 10(-6) M fluorescein solution at pH 12.25 containing 0.0005% (w/v) hexadimethrine bromide was used as background electrolyte. Quality parameters such as run-to-run, day-to-day precision and limits of detection were calculated, and better figures of merit were obtained for the coelectrooosmotic conditions than for the counterelectroosmotic mode. The coelectroosmotic method was applied to the quantitation of the hexosamine contents in glycosaminoglycans after acid hydrolysis. The method proved to be suitable for the determination of dermatan sulfate in heparin down to 2% (w/w).     

Proton nuclear magnetic resonance characterisation of glycosaminoglycans using chemometric techniques

Various types of glycosaminoglycans (GAGs) including heparins, chondroitin sulfates, dermatan sulfate and hyaluronic acid were studied from their proton nuclear magnetic resonance (1H NMR) spectra using chemometric techniques. Despite the complexity of the 1H NMR signals, data analysis using principal component analysis enabled the different GAG classes to be distinguished and permitted their classification according to their chemical structure. The analysis of the composition of the major disaccharide unit and other relevant chemical structures in the heparin samples was performed using partial least squares regression.