Occurrence and risk assessment of pesticides and pharmaceuticals in viticulture impacted watersheds from Northwest Spain.

An automated analytical methodology was developed, validated and applied to monitor 73 organic pollutants (pesticides and pharmaceuticals) in surface and groundwater samples obtained in watersheds from an intensive viticulture, rural region, in the Northwest of Spain. Filtered samples were concentrated using a reusable solid-phase extraction sorbent, on-line combined with liquid chromatography tandem mass spectrometry (LC-MS/MS). The analytical procedure achieved limits of quantification between 1 ng L-1 and 10 ng L-1, with a throughput of 2 samples hour-1, providing accurate recoveries for more than 90% of the 73 selected compounds, using calibration solutions prepared in ultrapure water (in presence of methanol and formic acid) as neat solvent. The distribution and the concentrations of pesticides in small streams impacted by discharges of treated municipal wastewaters were different in rural and residential areas. On the other hand, pharmaceuticals showed a similar distribution in both streams. In surface waters from viticulture impacted watersheds, with a limited influence of municipal wastewaters, pulses of pesticides were noticed, with values above 100 ng L-1 for several fungicides. Cardiovascular pharmaceuticals, psychiatric drugs and/or their transformation products were also ubiquitous in these samples, with low, but relatively stable concentrations among sampling campaigns. Within the suite of investigated compounds, maximum pesticide residues remained below their predicted-non effect concentration (PNEC) in all samples. On the other hand, the environmental concentrations of the cardiovascular drug olmesartan stayed systematically above its PNEC in fresh water samples.

Approaches to liquid chromatography tandem mass spectrometry assessment of glyphosate residues in wine.

The performance of two different analytical methodologies to investigate the presence of glyphosate (GLY) and aminomethylphosphonic acid (AMPA) residues in wine samples was evaluated. Transformation of compounds in their fluorene-9-methyloxycarbonyl derivatives permitted their separation under reversed-phase liquid chromatography with tandem mass spectrometry (LC-MS/MS) determination. Although the wine matrix severely impaired the efficiency of GLY derivatization, this drawback was solved using a molecularly imprinted sorbent for the previous, selective extraction of GLY and AMPA from wine. Alternatively, the use of a strong anionic exchange, polyvinyl alcohol-based LC column, turned to be the most effective alternative for direct determination of both compounds in diluted wine samples. The chromatographic behavior of this column and the magnitude of matrix effects observed during analysis of diluted wine samples were significantly affected by the composition of the mobile phase. Under final working conditions, this column permitted the separation of AMPA and the fungicide fosetyl (which shows common transitions in tandem MS/MS methods), it improved significantly the sample throughput versus extraction-derivatization-purification method, and it allowed the use of solvent-based calibration standards. Both analytical procedures provided similar limits of quantification (LOQs) for GLY (0.5-1.0 ng mL-1), while the multistep method was 8 times more sensitive to AMPA than the direct procedure. GLY residues stayed above method LOQs in 70% of the processed wines; however, concentrations measured in 95% of positive samples remained 100 times below the maximum residue limit (MRL) set for GLY in vinification grapes.

Assessment of direct analysis in real time accurate mass spectrometry for the determination of triclosan in complex matrices.

In this work, the applicability of direct analysis in real time coupled to accurate mass spectrometry (DART-MS) to the quantitative determination of triclosan (TCS) in samples with increasing complexity, from personal care products to extracts from sewage, is investigated. In the first term, DART-MS spectra of TCS as free phenol and as derivatized species are characterized; thereafter, the effects of several instrumental variables in the detectability of TCS (i.e., temperature, solvent, and compound holder) are discussed. Under final selected conditions, TCS was determined from its [M-H]- ions, without need of derivatization, attaining an instrumental limit of quantification of 5 ng mL-1, with a linear response range up to 1000 ng mL-1. Complex matrices, such as solid-phase extracts obtained from environmental water samples, moderately inhibited the ionization efficiency of TCS, with signal attenuation percentages in the range of 6 to 57%, depending on the sample type and on the concentration factor provided by the SPE procedure. The accuracy of results obtained by DART-MS was evaluated using liquid chromatography (LC) with MS detection; in both cases, a time-of-flight (TOF) MS instrument was employed for the selective determination of the [M-H]- ions of TCS (m/z values 286.9439 and 288.9410) using a mass window of 20 ppm. DART-MS did not only provide enough sensitivity to detect the presence of TCS in environmental samples (raw and treated wastewater as well as freeze-dried sludge), but also measured concentrations matched those determined by LC-ESI-TOF-MS, with only slightly higher standard deviations. During analysis of personal care products, containing much higher concentrations of TCS in a less complex matrix, both techniques were equivalent in terms of accuracy and precision. Graphical abstract.

Assessment of UV combined with free chlorine for removal of valsartan acid from water samples.

Valsartan acid (VALA) is a persistent and mobile pollutant, ubiquitously distributed in the aquatic environment. Herein, we assessed the efficiency of UV/free chlorine for the removal of this pollutant. Degradation experiments were performed using different water samples, considering several pH values and concentrations of inorganic anions. Time-course of VALA was measured by injection of different reaction time aliquots in a liquid chromatography (LC) triple quadrupole (QqQ) mass spectrometry (MS) system, whilst the study of potential transformation products (TPs) was evaluated by LC combined with a hybrid quadrupole time-of-flight (QTOF) MS system. Formation of volatile disinfection by-products (DBPs) was investigated by gas chromatography (GC) with TOF-MS detection. Compared to free chlorine treatment and UV photolysis, the combination of both parameters significantly enhanced the degradability of VALA. At neutral pH, UV/free chlorine was also more effective than UV/H2O2 to remove VALA from spiked water solutions. Three TPs of VALA were tentatively identified by LC-QTOF-MS, although only one was stable in the UV/free chlorine media. As regards volatile DBPs, the formation of chloroform, dichloroacetonitrile, di- and trichloroacetic acid was noticed. The mass yield of DBPs formation from VALA varied from 0.3% (dichloroacetonitrile) to 1.1% (chloroform). The efficiency of UV/free chlorine was first investigated in spiked solutions with increasing complexities: ultrapure, river and treated wastewater. Thereafter, the feasibility of reducing VALA levels in polluted river water was demonstrated.

Robustness assessment in computer-assisted liquid chromatography procedures based on desirability functions.

A novel approach based on the use of desirability functions is presented for the robustness assessment of liquid chromatographic separations as derived from computer-assisted methods development processes. The approach is based on generally accepted hypothesis that a robust separation procedure will be inert to small random variations of the operational variables, typically encountered in the day-to-day routine analytical practice. This means that peak positions along the chromatograms must keep standstill or move insignificantly when operational variables are not intentionally changed. Thus, the degree of peak positions variation as evaluated from mathematical retention models can be used to assess the robustness of the developed procedures before testing the actual performance experimentally. In the approach proposed, this assessment is obtained by fixing a bilateral partial desirability window around each peak in the simulated chromatogram. The whole chromatogram robustness is characterized by an overall desirability value calculated as the geometric mean of the partial desirability windows evaluation. An added advantage of this approach is that the robustness value calculated is normalized between zero and one and thus, easy to interpret. Thus, when chromatograms are simulated and small random variations are introduced into the operational factors of the model, values for the overall desirability close to one means that the procedure performs robustly. On the contrary, low values for the overall desirability clearly indicated a serious lack of robustness. When used in conjunction with the Pareto optimality approach, as shown here, this robustness assessment strategy allows testing several Pareto front solutions before the final experimental testing which is always needed. In this way, a dramatical reduction of the experimental effort is obtained. Although the approach is theoretically applicable to any chromatographic separation, examples of reversed phase liquid chromatographic procedures are used to show the performance of the proposed methodology.

Assessment of gas chromatography time-of-flight mass spectrometry for the screening of semi-volatile compounds in indoor dust.

Indoor dust contains a complex mixture of anthropogenic and synthetic compounds closely related to dermal and respiratory diseases. Target methods have been developed for the quantification of distinct groups of substances in dust samples; however, the comprehensive characterization of the different species existing in this matrix remains a challenging issue. Herein, we assess the performance of gas chromatography (GC) time-of-flight mass spectrometry (TOF-MS), using electron ionization (EI), for the screening of compounds present in indoor dust. Samples are processed by pressurized-liquid extraction (PLE) before GC-EI-TOF-MS analysis. The study proposes a data mining workflow for the non-target identification of species contained in dust extracts, aided by preliminary comparison with nominal resolution EI-MS spectra in the NIST17 library. The possibilities, and the limitations, of the above approach are discussed and the identities of >75 compounds are confirmed by comparison with authentic standards in dust from indoor environments. Some species, such as indigo, phthalic anhydride, 2,4-toluene di-isocyanate, phthalimide, certain UV absorbers and octyl isothiazolinone, identified in this research, have not been previously considered in target methods dealing with dust analysis. The study also evaluates two different algorithms for the suspected-target screening of dust extracts using a customized library of accurate EI-MS spectra. Finally, a semi-quantitative estimation of the range of concentrations for a group of 44 pollutants in a set of 27 dust samples is provided.

Free chlorine reactions of angiotensin II receptor antagonists: Kinetics study, transformation products elucidation and in-silico ecotoxicity assessment.

Angiotensin II receptor antagonists (ARA II) are widely employed in the treatment of hypertension-related diseases. Because of their partial metabolization and limited biodegradability, these drugs have become ubiquitous pollutants in the aquatic environment, including surface water. This research evaluated the reactivity of the ARA II drugs: irbesartan (IRB), losartan (LOS) telmisartan (TEL) and valsartan (VAL) with free chlorine. Responses of parent compounds and their transformation products (TPs) were followed by liquid chromatography (LC) with quadrupole (Q) time-of-flight (TOF) mass spectrometry. Degradation experiments were carried out using ultrapure and river water samples, adjusted at different pHs and, in some cases, adding a small amount (ng mL-1 level) of bromide salts. Whilst TEL and VAL remained stable in presence of relatively high concentrations of free chlorine (10 mg L-1), IRB and LOS were removed according to a pseudo-first order kinetics model. Considering an initial chlorine concentration of 10 mg L-1, their half-lives varied between 6 and 734 min, depending mostly on the water pH. IRB reacted with free chlorine through hydroxylation processes, with and without molecular cleavage and re-arrangements in the imidazolone ring. Its TPs showed a lower in-silico predicted toxicity than the parent drug. In case of LOS, two major competitive degradation routes were identified. They involved replacement of the methanol group attached to the imidazole cycle by chlorine or bromine, and the cleavage of this cycle with removal of the chlorinated carbon and the nitrogen in alpha position. The TPs generated following the first route are predicted to be more toxic than LOS.

Assessment of dispersive liquid-liquid microextraction conditions for gas chromatography time-of-flight mass spectrometry identification of organic compounds in honey.

The suitability of the dispersive liquid-liquid microextraction (DLLME) technique for gas chromatography (GC) characterization of minor organic compounds in honey samples is evaluated. Under optimized conditions, samples were pre-treated by liquid-liquid extraction with acetonitrile followed by DLLME using carbon tetrachloride (CCl4, 0.075 mL) as extractant. The yielded settled phase was analyzed by GC using high resolution time-of-flight (TOF) mass spectrometry (MS). The whole sample preparation process is completed in approximately 10 min, with a total consumption of organic solvents below 4 mL, relative standard deviations lower than 12% and with more than 70 organic compounds, displaying linear retention index in the range from 990 to 2900, identified in the obtained extracts. In comparison with HS SPME extraction, higher peak intensities were attained for most volatile and semi-volatile compounds amenable to both extraction techniques. Furthermore, other species such as highly polar and water soluble benzene acids, long chain fatty acids, esters and flavonoids, which are difficult to concentrate by HS SPME, could be identified in DLLME extracts. Some of the compounds identified in DLLME extracts have been proposed as useful for samples classification and/or they are recognized as markers of honeys from certain geographic areas.

Assessment of gas chromatography time-of-flight accurate mass spectrometry for identification of volatile and semi-volatile compounds in honey.

The performance of gas chromatography (GC) combined with a hybrid quadrupole time-of-flight (QTOF) mass spectrometry (MS) system for the determination of volatile and semi-volatile compounds in honey samples is evaluated. After headspace (HS) solid-phase microextraction (SPME) of samples, the accurate mass capabilities of the above system were evaluated for compounds identification. Accurate scan electron impact (EI) MS spectra allowed discriminating compounds displaying the same nominal masses, but having different empirical formulae. Moreover, the use of a mass window with a width of 0.005 Da provided highly specific chromatograms for selected ions, avoiding the contribution of interferences to their peak areas. Additional information derived from positive chemical ionization (PCI) MS spectra and ion product scan MS/MS spectra permitted confirming the identity of novel compounds. The above possibilities are illustrated with examples of honey aroma compounds, belonging to different chemical classes and containing different elements in their molecules. Examples of compounds whose structures could not be described are also provided. Overall, 84 compounds, from a total of 89 species, could be identified in 19 honey samples from 3 different geographic areas in the world. The suitability of responses measured for selected ions, corresponding to above species, for authentication purposes is assessed through principal components analysis.

Assessment of silicone as support to investigate the transformation routes of organic chemicals under environmental conditions and UV exposure. Application to selected fungicides.

The suitability of bulk silicone as support to follow the degradation of chemical compounds under environmental conditions and UV radiation is illustrated selecting three fungicides (fenhexamid, FEN; triadimenol, TRI and difenoconazole, DIF) as model compounds. These precursor species were first absorbed in silicone supports (10 mm length × 2 mm i.d. and 0.5 mm thickness) and then kept outdoors for several days (up to 2 months) or exposed to UV radiation (254 nm), from a low pressure mercury lamp, in the laboratory. Degradation of precursor fungicides and by-products formation was followed by liquid chromatography (LC) quadrupole time-of-flight (QTOF) mass spectrometry (MS), after desorption of silicone supports using 0.5 mL of acetonitrile. Half-lives (t(1/2)) measured under UV exposure varied from 5 to 100 min. As regards environmental conditions, the most stable fungicide was DIF, degraded by just 15 % after 2 months; whereas, t(1/2) values of 30 and 83 h were calculated for FEN during summer and autumn, respectively. Supports contained by-products arising from precursor species through de-chlorination, cleavage, hydroxylation, intra-molecular cyclation and oligomerization reactions. Most of them have been previously identified in soil surface, vegetable leaves and water after application of fungicides in agriculture fields. The low cost of silicone tubes (ca. 0.4 Euros), added to their excellent chemical stability and capability to retain precursor species and their by-products, make them ideal supports to follow the transformation routes of organic compounds under environmental and simulated conditions, even for relatively stable species with t(1/2) in the range of weeks or months.

Assessment of benzophenone-4 reactivity with free chlorine by liquid chromatography quadrupole time-of-flight mass spectrometry.

The stability of the UV filter benzophenone-4 (BP-4) in free chlorine-containing water was investigated, for the first time, by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QqTOF-MS). High mass accuracy and resolution capabilities of this hybrid mass spectrometer were used for the reliable assignation of empirical formulae and chemical structures of BP-4 derivatives. Time-course profiles of the parent compound and its by-products were simultaneously recorded by direct injection of sample aliquots, after quenching the excess of chlorine, in the LC-QqTOF-MS system. At neutral pHs, in excess of chlorine, BP-4 showed a limited stability fitting a pseudo-first-order degradation kinetics. A noticeable reduction in the half-lives of BP-4 was observed when increasing the sample pH between 6 and 8 units and also in presence of bromide traces. The reaction pathway of this UV filter involved a first electrophilic substitution of hydrogen per chlorine (or bromide) in the phenolic ring, followed by oxidation of the carbonyl moiety to an ester group, which induced a further electrophilic substitution in the same aromatic ring. Above reactions were also noticed when mixing a BP-4 containing personal care product with chlorinated tap water and in chlorinated swimming pool and sewage water, previously spiked with a BP-4 standard.

Fast throughput, highly sensitive determination of allergenic disperse dyes in textile products by use of sample composition.

A simple, highly sensitive and fast procedure for the control of allergenic disperse dyes in textile products was optimized. The method is based on ultrasound assisted extraction of textile samples with 20 mL of methanol under controlled conditions (15 min, 70 degrees C) followed by separation and analysis by LC-MS-MS. The sample preparation process was optimized by means of a surface response experimental design and provided quantitative recoveries of dyes, much better than the poor recoveries provided by current standard procedures. The chromatographic separation was optimized by means of computer-assisted method development by use of a special chemometric tool developed specifically for LC-MS systems, as previously reported by the authors. The result is a rapid chromatographic procedure that enables accurate quantification, at very low concentrations, of all 23 allergenic and/or carcinogenic disperse dyes considered. Matrix effects in the LC-MS procedure were studied. Under the experimental conditions, both conventional and strategic sample composition are proposed as efficient procedures that reduce the costs and work involved in the control of allergenic dyes in finished textile products. The benefits of strategic sample composition are demonstrated by means of an example case study, and the pros and cons of preparing the composite samples from sample extracts or directly from textile products are discussed.

Dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometry for the rapid and sensitive determination of UV filters in environmental water samples.

The performance of the dispersive liquid-liquid microextraction (DLLME) technique for the determination of eight UV filters and a structurally related personal care species, benzyl salicylate (BzS), in environmental water samples is evaluated. After extraction, analytes were determined by gas chromatography combined with mass spectrometry detection (GC-MS). Parameters potentially affecting the performance of the sample preparation method (sample pH, ionic strength, type and volume of dispersant and extractant solvents) were systematically investigated using both multi- and univariant optimization strategies. Under final working conditions, analytes were extracted from 10 mL water samples by addition of 1 mL of acetone (dispersant) containing 60 µL of chlorobenzene (extractant), without modifying either the pH or the ionic strength of the sample. Limits of quantification (LOQs) between 2 and 14 ng L(-1), inter-day variability (evaluated with relative standard deviations, RSDs) from 9% to 14% and good linearity up to concentrations of 10,000 ng L(-1) were obtained. Moreover, the efficiency of the extraction was scarcely affected by the type of water sample. With the only exception of 2-ethylhexyl-p-dimethylaminobenzoate (EHPABA), compounds were found in environmental water samples at concentrations between 6 ± 1 ng L(-1) and 26 ± 2 ng mL(-1).

Microwave-assisted extraction and large-volume injection gas chromatography tandem mass spectrometry determination of multiresidue pesticides in edible seaweed.

A microwave-assisted extraction method followed by clean-up with solid-phase extraction (SPE) combined with large-volume injection gas chromatography-tandem mass spectrometry (LVI-GC-MS/MS) for the analysis of 17 pesticides in wild and aquaculture edible seaweeds has been developed. An experimental central composite design was employed to evaluate the effects of the main variables potentially affecting the extraction (temperature, time, and solvent volume) and to optimize the process. The most effective microwave extraction conditions were achieved at 125 °C and 12 min with 24 mL of hexane/ethyl acetate (80:20). SPE clean-up of the extracts with graphitized carbon and Florisil, optimized by means of the experimental design, proved to be efficient in the removal of matrix interferences. The analytical recoveries were close to 100% for all the analytes, with relative standard deviations lower than 13%. The limits of detection ranged from 0.3 to 23.1 pg g(-1) and the limits of quantification were between 2.3 and 76.9 pg g(-1), far below the maximum residue levels established by the European Union for pesticides in seaweed. The results obtained prove the suitability of the microwave-assisted extraction for the routine analysis of pesticides in aquaculture and wild seaweed samples.