Exploration of environmental contaminants in honeybees using GC-TOF-MS and GC-Orbitrap-MS.

This study reports an analytical approach by gas chromatography and high-resolution mass spectrometry (HRMS) intended to be used for investigation of non-targeted environmental contaminants in honeybees. The approach involves a generic extraction and analysis with two GC-HRMS systems: time-of-flight and Orbitrap analyzers, GC-TOF-MS, and GC-Orbitrap-MS operated in electron-impact ionization (EI) mode. The workflow for screening of non-targeted contaminants consisted of initial peak detection by deconvolution and matching the first-stage mass spectra EI-MS with a nominal mass spectral library. To gain further confidence in the structural characterization of the contaminants under investigation, molecular formula of representative ions (molecular and fragment ions) was provided for those with an accurate mass scoring (error < 5 ppm). This methology was applied for screening environmental contaminants in 75 samples of adult honeybee. This approach has provided the tentative identification of environmental contaminants belonging to different chemical groups, among them, PAHs, phthalates and synthetic musks. Residues of veterinary treatments used in apiculture were also detected in the honeybee samples.

Identification and measurement of veterinary drug residues in beehive products.

There is an increasing concern about the negative impacts of veterinary drugs in beehive compartments. This study evaluates the presence and distribution of chemical residues in beeswax, bee bread and honey and determinates in what extension honeybees are exposed to them. Samples were analyzed by LC-MS/MS and GC-MS/MS with a wide scope of 322 chemical residues. Samples were collected from apiaries located in rural and forest areas, showing no difference in contamination of phytosanitary applications. Residues of acaricides used for sanitary treatments, coumaphos and two transformation products of amitraz (DMF and DMPF), were quantified at higher levels in wax and bee bread than in honey in most cases. Coumaphos, DMF and DMPF were detected in honey in the range 6-36 µg.kg-1; 45-541 µg.kg-1; 15-107 µg.kg-1, respectively. All, except one sample, were below the EU MRLs, 396/2005 Regulation. Other pesticide residues were detected in beeswax and bee bread at various levels.

Non-target evaluation of contaminants in honey bees and pollen samples by gas chromatography time-of-flight mass spectrometry.

This work presents a non-targeted screening approach for the detection and quantitation of contaminants in bees and pollen, collected from the same hive, by GC-EI-ToF-MS. It consists of a spectral library datasets search using a compound database followed by a manual investigation and analytical standard confirmation together with semi-quantitation purposes. Over 20% of the compounds found automatically by the library search could not be confirmed manually. This number of false positive detections was mainly a consequence of an inadequate ion ratio criterion (±30%), not considered in the automatic searching procedure. Eight compounds were detected in bees and pollen. They include insecticides/acaricides (chlorpyrifos, coumaphos, fluvalinate-tau, chlorfenvinphos, pyridaben, and propyl cresol) at a concentration range of 1-1207 µg kg-1, herbicides (oxyfluorfen) at a concentration range of 212-1773 µg kg-1 and a growth regulator hormone (methoprene). Some compounds were detected only in pollen; such as herbicides (clomazone), insecticides/acaricides and fungicides used to control Varroa mites as benzylbenzoate, bufencarb, allethrin, permethrin, eugenol and cyprodinil. Additional compounds were detected only in bees: flamprop-methyl, 2-methylphenol (2-49 µg kg-1) and naphthalene (1-23 µg kg-1). The proposed method presents important advantages as it can avoid the use of an unachievable number of analytical standards considered target compounds "a priori" but not present in the analyzed samples.

Identification of non-intentionally added substances in food packaging nano films by gas and liquid chromatography coupled to orbitrap mass spectrometry.

The control of chemical migration from new functionalized food contact materials (FCMs) is a challenge for meeting food safety requirements. The non-intentionally added substances (NIAS) constitute a group of chemicals that are not applied, but may be introduced or formed during the production process of FCMs. This study describes a multi-analytical approach for the evaluation of unknown substances that migrate from FCMs. A case study is presented using a developed polymer consisting of a monolayer film with polylactic acid (PLA), polylimonene (PL) and zinc oxide nanoparticles (ZnO NPs). This approach incorporates the platforms of ICP-MS (inductively coupled plasma mass spectrometry), to determine whether there is transference of ZnO NPs used as antimicrobial agent and, the systems GC-MS and LC-MS (gas / liquid chromatography coupled to a quadrupole orbitrap mass spectrometer) for the characterization of the chemical structure of NIAS using the molecular mass and specific features of mass fragmentation. The screening of unknown compounds comprised retrospective analysis and data processing using both, a mass spectral library and databases, for GC and LC data, respectively. This approach has provided the tentative identification and quantification of seven NIAS, 3 by GC (Tripropylene glycol diacrylate, 10-Heneicosene and α-Tocopherol acetate) and 4 by LC (N,N-Diethyldodecanamide, N-[(9Z)-9-Octadecen-1-yl]acetamide, 1-Palmitoylglycerol and Glycerol stearate). This migration study was carried out according to the standard protocols established by EU regulation for FCMs.

Determination of selected environmental contaminants in foraging honeybees.

Colony losses of honeybees have been of great concern in the last years. To explain these losses, several studies have been reported, and various factors, such as pathogens and pesticides, have been considered as possible causes. Nevertheless, organic contaminants, rather than pesticides, are continuously released to the environment, and can be intercepted by honeybees during foraging with the possible consequent damage. Azoles and organophosphorus esters have been selected in this work as environmental contaminants to be monitored in honeybees. A fast and robust method has been developed to determine these organic pollutants in honeybees. It is based on matrix solid phase dispersion (MSPD), which performs sample dispersion with extraction and clean up in the same step, followed by LC-ESI-MS/MS determination. Recoveries of the method varied between 73% and 119% and MQLs ranged from 0.8 to 4 ng g(-1). Honeybee samples from ten apiaries located in different regions were analyzed applying the developed method. Azole compounds were found at low levels, but not in all samples, while organophosphorus esters were found in most samples whatever location. Tris-(2-chloroisopropyl) phosphate, TCPP, and tributyl phosphate, TBP, were detected in all honeybees samples at levels higher than the rest of organophosphates analyzed.

Screening of environmental contaminants in honey bee wax comb using gas chromatography-high-resolution time-of-flight mass spectrometry.

This study reports an analytical approach intended to be used for investigation of non-targeted environmental contaminants and to characterize the organic pollution pattern of bee wax comb samples. The method comprises a generic extraction followed by detection with gas chromatography coupled to high-resolution time-of-flight mass spectrometry (GC-TOF-MS), operated in electron impact ionization (EI) mode. The screening approach for the investigation of non-targeted contaminants consisted of initial peak detection by deconvolution and matching the first-stage mass spectra EI-MS(1) with a nominal mass spectral library. To gain further confidence in the structural characterization of the contaminants under investigation, the molecular formula of representative ions (molecular ion when present in the EI spectrum) and, for at least other two fragment ions, was provided for those with an accurate mass scoring (mass error < 5 ppm). This methodology was applied for screening environmental contaminants in 50 samples of bee wax comb. This approach has allowed the tentative identification of some GC-amenable contaminants belonging to different chemical groups, among them, phthalates and polycyclic aromatic hydrocarbons (PAHs), along with residues of veterinary treatments used in apiculture.

Simultaneous screening of targeted and non-targeted contaminants using an LC-QTOF-MS system and automated MS/MS library searching.

Simultaneous high-resolution full-scan and tandem mass spectrometry (MS/MS) analysis using time of flight mass spectrometry brings an answer for increasing demand of retrospective and non-targeted data analysis. Such analysis combined with spectral library searching is a promising tool for targeted and untargeted screening of small molecules. Despite considerable extension of the panel of compounds of tandem mass spectral libraries, the heterogeneity of spectral data poses a major challenge against the effective usage of spectral libraries. Performance evaluation of available LC-MS/MS libraries will significantly increase credibility in the search results. The present work was aimed to evaluate fluctuation of MS/MS pattern, in the peak intensities distribution together with mass accuracy measurements, and in consequence, performance compliant with ion ratio and mass error criteria as principles in identification processes for targeted and untargeted contaminants at trace levels. Matrix effect and ultra-trace levels of concentration (from 50 ng l(-1) to 1000 ng l(-1) were evaluated as potential source of inaccuracy in the performance of spectral matching. Matrix-matched samples and real samples were screened for proof of applicability. By manual review of data and application of ion ratio and ppm error criteria, false negatives were obtained; this number diminished when in-house library was used, while with on-line MS/MS databases 100% of positive samples were found. In our experience, intensity of peaks across spectra was highly correlated to the concentration effect and matrix complexity. In turn, analysis of spectra acquired at trace concentrations and in different matrices results in better performance in providing correct and reliable identification.

Post-acquisition data processing for the screening of transformation products of different organic contaminants. Two-year monitoring of river water using LC-ESI-QTOF-MS and GCxGC-EI-TOF-MS.

This study describes a comprehensive strategy for detecting and elucidating the chemical structures of expected and unexpected transformation products (TPs) from chemicals found in river water and effluent wastewater samples, using liquid chromatography coupled to electrospray ionization quadrupole-time-of-flight mass spectrometer (LC-ESI-QTOF-MS), with post-acquisition data processing and an automated search using an in-house database. The efficacy of the mass defect filtering (MDF) approach to screen metabolites from common biotransformation pathways was tested, and it was shown to be sufficiently sensitive and applicable for detecting metabolites in environmental samples. Four omeprazole metabolites and two venlafaxine metabolites were identified in river water samples. This paper reports the analytical results obtained during 2 years of monitoring, carried out at eight sampling points along the Henares River (Spain). Multiresidue monitoring, for targeted analysis, includes a group of 122 chemicals, amongst which are pharmaceuticals, personal care products, pesticides and PAHs. For this purpose, two analytical methods were used based on direct injection with a LC-ESI-QTOF-MS system and stir bar sorptive extraction (SBSE) with bi-dimensional gas chromatography coupled with a time-of-flight spectrometer (GCxGC-EI-TOF-MS).

Investigation of Galaxolide degradation products generated under oxidative and irradiation processes by liquid chromatography/hybrid quadrupole time-of-flight mass spectrometry and comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry.

RATIONALE: Polycyclic musks have become a concern due to their bioaccumulation potential and ecotoxicological effects. The HHCB transformation product (TP) (1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethyl-cyclopenta[γ]-2-benzopyran; HHCB-lactone) is the most stable intermediate generated and it is frequently detected in river waters. The aim of this work was the identification of relevant TPs generated from UV irradiation and ozone treatments. METHODS: Identification of HHCB TPs was carried out by liquid chromatography/hybrid quadrupole time-of-flight mass spectrometry (LC/ESI-QTOF-MS) and two-dimensional gas chromatography/electron impact time-of-flight mass spectrometry (GC×GC-EI-TOF-MS). With LC/ESI-QTOF-MS, TPs were characterized by means of mass accuracy in both full-scan and MS/MS modes through information-dependent acquisition (IDA) and direct injection on-column. With stir bar sorptive extraction (SBSE)-GC×GC-EI-TOF-MS, identification was based on the enhanced separation capacity and screening of unknowns through the acquisition of full-range mass spectra. RESULTS: The effectiveness of these complementary techniques allowed a detailed evaluation of the main TPs. Eighteen TPs were elucidated based on mass accuracy, in both full-scan and MS/MS modes using LC/ESI-QTOF-MS with mass errors below 5 ppm and 10 ppm (mostly), respectively. Most of the TPs had not been analytically identified in previous studies. Separation of the enantiomeric species (R) and (S) of HHCB-lactone, and the identification of other relevant TPs, was performed using SBSE-GC×GC-EI-TOF-MS. CONCLUSIONS: LC/ESI-QTOF-MS and GC×GC-EI-TOF-MS analysis provides the best alternative for TP identification of chemicals of concern, which have a wide range of polarities and isobaric compounds. A prediction of PBT (persistence, bioaccumulation and toxicity) using the PBT Profiler program suggested a classification of 'very persistent' and 'very toxic' for most of the TPs identified.

Qualitative and quantitative analysis of poly(amidoamine) dendrimers in an aqueous matrix by liquid chromatography-electrospray ionization-hybrid quadrupole/time-of-flight mass spectrometry (LC-ESI-QTOF-MS).

This work introduces a liquid chromatography-electrospray ionization-hybrid quadrupole/time-of-flight mass spectrometry (LC-ESI-QTOF-MS)-based method for qualitative and quantitative analysis of poly(amidoamine) (PAMAM) dendrimers of generations 0 to 3 in an aqueous matrix. The multiple charging of PAMAM dendrimers generated by means of ESI has provided key advantages in dendrimer identification by assignation of charge state through high resolution of isotopic clusters. Isotopic distribution in function of abundance of isotopes (12)C and (13)C yielded valuable and complementarity data for confident characterization. A mass accuracy below 3.8 ppm for the most abundant isotopes (diagnostic ions) provided unambiguous identification of PAMAM dendrimers. Validation of the LC-ESI-QTOF-MS method and matrix effect evaluation enabled reliable and reproducible quantification. The validation parameters, limits of quantification in the range of 0.012 to 1.73 µM, depending on the generation, good linear range (R > 0.996), repeatability (RSD < 13.4%), and reproducibility (RSD < 10.9%) demonstrated the suitability of the method for the quantification of dendrimers in aqueous matrices (water and wastewater). The added selectivity, achieved by multicharge phenomena, represents a clear advantage in screening aqueous mixtures due to the fact that the matrix had no significant effect on ionization, with what is evidenced by an absence of sensitivity loss in most generations of PAMAM dendrimers. Fig Liquid chromatography-electrospray ionization-hybrid quadrupole/time of flight mass spectrometry (LC-ESI-QTOF-MS) based method for qualitative and quantitative analysis of PAMAM dendrimers in aqueous matrix.

Simultaneous measurement in mass and mass/mass mode for accurate qualitative and quantitative screening analysis of pharmaceuticals in river water.

A new approach for the analysis of pharmaceuticals (target and non-target) in water by LC-QTOF-MS is described in this work. The study has been designed to assess the performance of the simultaneous quantitative screening of target compounds, and the qualitative analysis of non-target analytes, in just one run. The features of accurate mass full scan mass spectrometry together with high MS/MS spectral acquisition rates - by means of information dependent acquisition (IDA) - have demonstrated their potential application in this work. Applying this analytical strategy, an identification procedure is presented based on library searching for compounds which were not included a priori in the analytical method as target compounds, thus allowing their characterization by data processing of accurate mass measurements in MS and MS/MS mode. The non-target compounds identified in river water samples were ketorolac, trazodone, fluconazole, metformin and venlafaxine. Simultaneously, this strategy allowed for the identification of other compounds which were not included in the library by screening the highest intensity peaks detected in the samples and by analysis of the full scan TOF-MS, isotope pattern and MS/MS spectra - the example of loratadine (histaminergic) is described. The group of drugs of abuse selected as target compounds for evaluation included analgesics, opioids and psychostimulants. Satisfactory results regarding sensitivity and linearity of the developed method were obtained. Limits of detection for the selected target compounds were from 0.003 to 0.01 µg/L and 0.01 to 0.5 µg/L, in MS and MS/MS mode, respectively - by direct sample injection of 100 µL.

In vitro dose-response effects of poly(amidoamine) dendrimers [amino-terminated and surface-modified with N-(2-hydroxydodecyl) groups] and quantitative determination by a liquid chromatography-hybrid quadrupole/time-of-flight mass spectrometry based method.

This article presents a dose-response study of the effects of two types of third-generation (G3) and fourth-generation poly(amidoamine) (PAMAM) dendrimers on two cell lines (RTG-2 and H4IIE) by in vitro cytotoxicity assays with 3-(4,5-dimethylthizol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), neutral red uptake (NRU), and lactate dehydrogenase (LDH) assays. We particularly investigated the potential cytotoxic effect of positive surface charge, which a cationic amino-terminated PAMAM dendrimer can display, on the marked ability of PAMAM dendrimers to cross the cell membrane compared with PAMAM dendrimers functionalized with chains of N-(2-hydroxydodecyl). Quantification of dose-response effects was performed by use of mass spectrometry analysis. The analytical method using liquid chromatography-hybrid quadrupole/time-of-flight mass spectrometry that we developed allowed characterization of defective dendrimers instead of "ideal structures." Identification was based on accurate mass measurement, assignment of elemental composition, and the fully resolved (13)C/(12)C isotopic clusters of the multiply charged ions of PAMAM dendrimers. Validation of the liquid chromatography-mass spectrometry method made possible reliable and reproducible quantification of the extracellular and intracellular concentration of dendrimers at a micromolar level (limits of detection from 0.14 to 1.34 µM and from 0.43 to 1.82 µM in standard and culture medium, respectively). A higher cytotoxicity was found with the H4IIE cell line for surface-modified PAMAM dendrimers. The LDH assay was significantly more sensitive than the MTT and NRU assays, with half-maximal inhibitory concentrations (IC(50)) of 12.96 and 38.31 µg mL(-1) for surface-modified G3 and G4 dendrimers, respectively. No cytotoxic effects, in terms of IC(50), of amino-terminated PAMAM dendrimers were observed on both H4IIE and RTG-2 cells when the concentration was below 500 µg mL(-1) for G3 and G4 dendrimers.

Occurrence and persistence of organic emerging contaminants and priority pollutants in five sewage treatment plants of Spain: two years pilot survey monitoring.

This work summarized all results obtained during almost two-years of a monitoring programme carried out in five municipal sewage treatment plants (STPs) located in the north, centre and south-east of Spain. The study evaluated the occurrence and persistence of a group of 100 organic compounds belonging to several chemical groups (pharmaceuticals, personal care products, pesticides and metabolites). The average removal efficiencies of the STPs studied varied from 20% (erythromycin) to 99% (acetaminophen). In analysed samples, we identified a large number of compounds at mean range concentrations between 7-59,495 ng/L and 5-32,720 ng/L for influent and effluent samples, respectively. This study also identified 20 of the mostly detected and persistent compounds in wastewater effluent, of which hydrochlorothiazide, atenolol, gemfibrozil, galaxolide and three metabolites (fenofibric acid, 4-AAA and 4-FAA), presented the highest average contribution percentages, in relation to the total load of contaminants for the different STPs effluent studied.

Development of a solvent-free method for the simultaneous identification/quantification of drugs of abuse and their metabolites in environmental water by LC-MS/MS.

This work details a rapid analytical method using direct sample injection for the simultaneous identification/quantification of 22 drugs of abuse, including some of their major metabolites, in environmental samples. This has been developed using a hybrid triple quadrupole-linear ion trap-mass spectrometer (QqLIT). With the increasing sensitivity of today's tandem mass spectrometers, direct injection analysis of water samples has become an attractive alternative to traditional analytical protocols, which often include a preliminary pre-concentration step. What's more, this kind of analysis is in accordance with many of the main objectives of so-called green analytical chemistry, or environmentally friendly practice. The analytical performance of the LC-MS/MS method was evaluated in three different water matrices (surface water, influent and effluent wastewater). Data acquisition was carried out in selected reaction monitoring (SRM) mode under time-scheduled conditions, monitoring two SRM transitions for simultaneous identification/quantification of all target compounds in the samples. Additionally, an experiment was performed using the information-dependent acquisition (IDA) scan to carry out the identification of those analytes for which the second transition was present at a low intensity. Finally, the two methodologies developed were applied to real samples for evaluation.

Evaluation of selected ubiquitous contaminants in the aquatic environment and their transformation products. A pilot study of their removal from a sewage treatment plant.

A simple method using direct sample injection combined with liquid chromatography tandem mass spectrometry has been developed for the simultaneous analysis of six alkaloid compounds in environmental samples. The target list includes two psychostimulants (nicotine and caffeine), three metabolites (cotinine, nicotinic acid and paraxanthine) and a coffee chemical (trigonelline). The analytical method was evaluated in three different matrices (surface water, influent and effluent wastewater). The method developed showed an adequate sensitivity, below 0.6 µg L(-1) for wastewater and 0.1 µg L(-1) for river matrices, without any prior treatment of the samples. Finally, the methodology was applied to real samples for evaluation of their removal from a sewage treatment plant and their persistence/fate in the aquatic environment. All compounds studied in this work were detected at all sampling points collected along the Henares River. However, nicotinic acid was only detected three times in treated sewage samples at levels above its detection limit.

Removal of pharmaceuticals and kinetics of mineralization by O(3)/H(2)O(2) in a biotreated municipal wastewater.

The ozonation of an effluent from the secondary clarifier of two Municipal Wastewater Treatment Plants was performed by using alkaline ozone and a combination of ozone and hydrogen peroxide. Alkaline ozonation achieved only a moderate degree of mineralization, essentially concentrated during the first few minutes; but the addition of hydrogen peroxide eventually led to a complete mineralization. The evolution of total organic carbon (TOC) as a measure of the extent of mineralization and the concentration of dissolved ozone were analyzed and linked in a kinetic model whose parameter represented the product of the exposure to hydroxyl radicals and the kinetic constant of indirect ozonation. This rate parameter yielded the highest values during the first part of O(3)/H(2)O(2) runs. The kinetic constant for the decomposition of ozone at the end of the run was also measured and computed for the non-oxidizable water matrix and yielded essentially the same values regardless of whether or not hydrogen peroxide was used. A group of 33 organic compounds, mainly pharmaceuticals and some relevant metabolites present in the wastewater effluents, were evaluated before and after the ozonation process using a liquid chromatography-hybrid triple-quadrupole linear ion trap system (LC-QqLIT-MS). The results demonstrate that the ozonation degrades these compounds with efficiencies of over 99% in most cases, even under low mineralization conditions in alkaline ozonation.

Identification of photocatalytic degradation products of bezafibrate in TiO(2) aqueous suspensions by liquid and gas chromatography.

In the present study the photocatalytic degradation of bezafibrate (BZF), a lipid regulator agent, has been investigated using TiO(2) suspensions and simulated solar light. The study focus on the identification of degradation products (DPs) using powerful analytical techniques such as liquid chromatography time of flight mass spectrometry (LC-TOF-MS), gas chromatography mass spectrometry (GC-MS), and high-performance liquid chromatography with diode-array detection (HPLC-DAD). Each technique provided complementary information that enabled the identification of 21 DPs. Accurate mass measurements obtained by LC-TOF-MS provided the elucidation of 17 DPs. Mass errors lower than 2mDa, allowed the assignment of empirical formula for the mayor DPs to be determined confidently. Three DPs were identified by GC-MS through the structural information provided by full scan mass spectra obtained by electron impact (EI) ionization and two more by HPLC-DAD by comparing the retention times (t(R)) and the UV spectra of the unknown DPs with those of commercial standards. Based on this by-product identification a possible multi-step degradation scheme was proposed. The pathways include single or multiple hydroxylation of BZF with subsequent phenoxy ring opening and the cleavage of the amide and ether bonds.

Evaluation of ozone-based treatment processes for wastewater containing microcontaminants using LC-QTRAP-MS and LC-TOF/MS.

This article describes the development of an enhanced liquid chromatography-mass spectrometry (LC-MS) method for the analysis of a selected group of 57 organic contaminants in wastewater. This group comprises 39 pharmaceuticals belonging to different therapeutical classes and 10 of their most frequent metabolites. Six pesticides and two disinfectants were also included. The LC-MS method was developed using a hybrid quadrupole/linear ion trap (Q TRAP) analyzer operating in selected reaction monitoring (SRM) mode (in both positive and negative electrospray ionization) in combination with a time-of flight (TOF) mass analyser. The application of both techniques provided very good results in terms of accurate quantification and unequivocal identification. Quantification was based on the use of a linearly accelerating (LINAC) high-pressure collision cell, which enable the analysis of a high number of compounds with enough acquisition data points for an optimal peak definition in SRM. Unequivocal identification was provided by the acquisition of at least two SRM transitions and by obtaining accurate mass measurements of the identified compounds with errors lower than 2 ppm. As an alternative for compounds where a second transition cannot be detected by Q-Trap-MS, the application of survey scans in enhanced product ion (EPI) was evaluated. The analytical performance of the method was evaluated in effluent wastewater samples. Linearity of response over three orders of magnitude was demonstrated (R2>0.99 for most compounds). Matrix effects resulting in suppression of the response were frequently observed, between 2-50% for most of compounds, except 4-DAA and 4-AA, which exhibit higher values (68%). Signal enhancement was also detected in 16 compounds. Method limits of detection (LOD) were between 0.1-50 ng L(-1). Finally, the methodology was successfully applied to the evaluation of the efficiency of two ozone-based treatments applied to the effluent from the secondary clarifier of a municipal wastewater treatment facility. Preliminary results are presented demonstrating that ozonation of wastewaters degrade pharmaceuticals with a high efficiency. Removals higher than 90% were reached for most of target analytes.

Photodegradation of malachite green under natural sunlight irradiation: kinetic and toxicity of the transformation products.

This article describes the photolytic degradation of malachite green (MG), a cationic triphenylmethane dye used worldwide as a fungicide and antiseptic in the aquaculture industry. Photolysis experiments were performed by direct exposure of a solution of MG in water to natural sunlight. The main transformation products (TPs) generated during the process were identified by liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS) and gas chromatography mass spectrometry (GC-MS). The 28 TPs identified with this strategy indicate that MG undergoes three main reactions, N-demethylation, hydroxylation and cleavage of the conjugated structure forming benzophenone derivatives. These processes involve hydroxyl radical attack on the phenyl ring, the N,N-dimethylamine group and the central carbon atom. The Vibrio fischeri acute toxicity test showed that the solution remains toxic after MG has completely disappeared. This toxicity could be assigned, at least in part, to the formation of 4-(dimethylamine)benzophenone, which has an EC(50,30 min) of 0.061 mg l(-1), and is considered "very toxic to aquatic organisms" by current EU legislation.

Application of high-performance liquid chromatography-tandem mass spectrometry with a quadrupole/linear ion trap instrument for the analysis of pesticide residues in olive oil.

This article describes the development of an enhanced liquid chromatography-mass spectrometry (LC-MS) method for the analysis of pesticides in olive oil. One hundred pesticides belonging to different classes and that are currently used in agriculture have been included in this method. The LC-MS method was developed using a hybrid quadrupole/linear ion trap (QqQ(LIT)) analyzer. Key features of this technique are the rapid scan acquisition times, high specificity and high sensitivity it enables when the multiple reaction monitoring (MRM) mode or the linear ion-trap operational mode is employed. The application of 5 ms dwell times using a linearly accelerating (LINAC) high-pressure collision cell enabled the analysis of a high number of pesticides, with enough data points acquired for optimal peak definition in MRM operation mode and for satisfactory quantitative determinations to be made. The method quantifies over a linear dynamic range of LOQs (0.03-10 microg kg(-1)) up to 500 microg kg(-1). Matrix effects were evaluated by comparing the slopes of matrix-matched and solvent-based calibration curves. Weak suppression or enhancement of signals was observed (<15% for most-80-of the pesticides). A study to assess the identification criteria based on the MRM ratio was carried out by comparing the variations observed in standard vs matrix (in terms of coefficient of variation, CV%) and within the linear range of concentrations studied. The CV was lower than 15% when the response observed in solvent was compared to that in olive oil. The limit of detection was < or =10 microg kg(-1) for five of the selected pesticides, < or =5 microg kg(-1) for 14, and < or =1 microg kg(-1) for 81 pesticides. For pesticides where additional structural information was necessary for confirmatory purposes-in particular at low concentrations, since the second transition could not be detected-survey scans for enhanced product ion (EPI) and MS3 were developed.