This paper provides scientific results from a European LIFE project carried out in the Valencian ... more This paper provides scientific results from a European LIFE project carried out in the Valencian region of Spain during the 2017 to 2018 time frame. In 2018, more than 60,000 tons of pesticides were commercialized in Spain, with approximately 15% destined for Valencian crops. In order to improve the air quality in the agricultural areas of this region, an innovative cropping system based on irrigation was developed and compared to conventional treatments based on hand-spray and turbo application. After applying conventional treatments to five types of crops (citrus, persimmon, nectarine, watermelon, and other stone fruits), a total of 13 active substances were detected in the air. The same active substances were applied to crops using the novel irrigation system, and no pesticide was detected in the air. Moreover, applicator and bystander populations in the region were assessed for their risk of inhalation exposure to pesticides, and no risk was found when either of the techniques, ...
The interest of ambient ionization mass spectrometry in clinical and forensic analysis has increa... more The interest of ambient ionization mass spectrometry in clinical and forensic analysis has increased in the last years as it allows the rapid and direct analysis of a wide variety of samples. Among the possible applications, the analysis in real-time of the exhaled breath has gained great attention, for the investigation of both endogenous compounds, as for example disease biomarkers, and exogenous compounds related to the consumption of certain products. Although commercial ionization sources are already available for breath analysis, they require an important economic investment and/or complex setups. In this article, we describe a new probe developed at our laboratory, fully compatible with the Waters Corp. atmospheric solids analysis probe (ASAP), and where manufacturing is neither expensive nor complex. This atmospheric breath analysis probe (ABAP) can be directly used in Waters mass spectrometry instruments without the need of modifying the ionization source, the instrument or the ASAP, being also compatible with the software provided by the manufacturer. The ionization of the compounds is based on atmospheric pressure chemical ionization. The prototype has been successfully used in a high-resolution mass spectrometry system, and applied to the analysis of compounds present in human breath, as well as to the tentative identification of different substances present in foods and reported in exhaled breath after food consumption. In addition, ABAP could also be compatible with other ionization techniques, such as photoionization.
Currently, the most powerful approach to monitor organic micropollutants (OMPs) in environmental ... more Currently, the most powerful approach to monitor organic micropollutants (OMPs) in environmental samples is the combination of target, suspect and non-target screening strategies using high resolution mass spectrometry (HRMS). However, the high complexity of sample matrices as well as the huge number of OMPs potentially present in samples at low concentrations pose an analytical challenge. Ion mobility separation (IMS) combined with HRMS instruments (IMS-HRMS) introduces an additional analytical dimension, providing extra information which facilitates the identification of OMPs. The collision cross section (CCS) value provided by IMS is unaffected by the matrix or chromatographic separation. Consequently, the creation of CCS databases and the inclusion of ion mobility within identification criteria are of high interest for an enhanced and robust screening strategy. In this work, a CCS library for IMS-HRMS, which is online and freely available, was developed for 556 OMPs in both positive and negative ionization modes using electrospray ionization. The inclusion of ion mobility data in widely adopted confidence levels for identification in environmental reporting is discussed. Illustrative examples of OMPs found in environmental samples are presented to highlight the potential of IMS-HRMS and to demonstrate the additional value of CCS data in various screening strategies.
The pesticide chlorpyrifos-methyl (CLP-m) has been identified in plant ingredients intended for a... more The pesticide chlorpyrifos-methyl (CLP-m) has been identified in plant ingredients intended for aquaculture feed production. To investigate systemic effects of CLP-m with emphasis on lipid metabolism, post-smolt Atlantic salmon were fed in duplicate (n=2) either diets with no CLP-m (Control) or CLP-m at different concentrations (0.1, 1.0 or 8.0 mg kg-1) for a total of 67 days (Low, Medium, High). Fish in all groups almost doubled their weight during the feeding trial from 262 ± 26g (mean ± SD) to 465 ± 64g (overall mean), with no significant effects on any growth parameters. There was a significant dosedependent inhibition of plasma cholinesterase activity (BuChE) after 67 days. The CLP-m biotransformation metabolite, TCP was detected in liver and bile, with low levels of the parent compound in the organs. Spleen somatic index decreased significantly with increasing dietary CLP-m intake. Hematocrit (%) decreased linearly with increasing dietary exposure to CLP-m after 30 days of exposure, but this decrease was less at 67 days of exposure. A significantly reduced content of arachidonic acid (ARA 20:4n-6), accompanied by a significantly increased content of the saturated fatty acid, palmitic acid (PA 16:0), was observed in liver phospholipids (PLs) with increasing dietary content of CLP-m. Major effects were seen on the PL classes in liver which showed a significantly decreased absolute content, possibly indicating inhibition of PL remodeling pathways or other membrane perturbation effects from CLP-m exposure. In conclusion, this study shows that the pesticide CLP-m is a relatively potent toxicant in Atlantic salmon, especially affecting liver PLs and ARA metabolism.
The potential of ultra-high-pressure liquid chromatography (UHPLC) coupled to hybrid quadrupole t... more The potential of ultra-high-pressure liquid chromatography (UHPLC) coupled to hybrid quadrupole timeof-flight mass spectrometry (QTOF-MS) for the screening and confirmation of antibiotics in water samples is illustrated in this paper. UHPLC presents several advantages over conventional liquid chromatography as it generates narrow peaks (increasing peak height and improving sensitivity) and reduces chromatographic runs. Regarding QTOF-MS, its increased mass resolution, high sensitivity in full-spectrum acquisition mode and mass accuracy, in both MS and MS/MS modes, make this technique ideal for the detection and reliable confirmation of organic contaminants in environmental samples. After a solidphase extraction using Oasis HLB cartridges, UHPLC-QTOF-MS has been applied in this work to several types of water samples (surface water and influent and effluent wastewaters). Several antibiotics were found in the samples, such as ofloxacin, ciprofloxacin, clarythromycin or erythromycin, among others. Moreover, the full spectrum data provided by TOF-MS acquisition has enabled searching for many other pharmaceuticals that could be present in the samples in a "post-target" way. This approach has allowed the detection and confirmation of paracetamol, omeprazole and codeine, among others. UPLC-QTOF-MS has been shown as an attractive and efficient hyphenated technique for the rapid detection and confirmation of pharmaceuticals in water with very little sample handling.
A fast and automated strategy has been developed for identifying unknown substances in the atmosp... more A fast and automated strategy has been developed for identifying unknown substances in the atmosphere (concretely, in the particulate matter, PM10) using LC-HRMS (MS3). A total of 15 samples were collected in three different areas (rural, urban and industrial). A sampling flow rate of 30 m3 h−1 was applied for 24 h, sampling a total volume of around 720 m3. A total of 49 compounds were tentatively identified using very restrictive criteria regarding exact mass, retention time, isotopic profile and both MS2 and MS3 spectra. Pesticides, pharmaceutical active compounds, drugs, plasticizers and metabolites were the most identified compounds. To verify whether the developed methodology was suitable, 11 substances were checked with their analytical standards and all of them were confirmed. Different profiles for industrial, rural and urban areas were examined. The Principal Component Analysis (PCA) model allowed us to separate the obtained data of the three assessed area. When the profile...
Synthetic cannabinoids receptor agonists (SCRAs) are often almost completely metabolised, and hen... more Synthetic cannabinoids receptor agonists (SCRAs) are often almost completely metabolised, and hence their pharmacokinetics should be carefully evaluated for determining the most adequate biomarker in toxicological analysis. Two structurally related SCRAs, AMB-FUBINACA and AMB-CHMICA, were selected to evaluate their in vivo metabolism and pharmacokinetics using male Sprague-Dawley rats. Brain, liver, kidney, blood (serum) and urine samples were collected at different times to assess the differences in metabolism, metabolic reactions, tissue distribution and excretion. Both compounds experimented O-demethyl reaction, which occurred more rapidly for AMB-FUBINACA. The parent compounds and O-demethyl metabolites were highly bioaccumulated in liver, and were still detected in this tissue 48 h after injection. The different indazole/indole N-functionalisation produced diverse metabolic reactions in this moiety and thus, different urinary metabolites were formed. Out of the two compounds, A...
The potential of high resolution mass spectrometry (HRMS) for the investigation of human in vivo ... more The potential of high resolution mass spectrometry (HRMS) for the investigation of human in vivo metabolism of 3,4-methylenedioxypyrovalerone (MDPV) using urine collected from a consumer (this is, in non-controlled experiments) has been investigated. As a control sample was not available, the common approach based on the comparison of a control/blank sample and samples collected after drug intake could not be used. Alternatively, an investigation based on common fragmentation pathways was applied, assuming that most metabolites share some fragments with the parent drug. An extension of this approach was also applied based on the fragmentation pathway of those metabolites identified in urine samples in the first step. The use of MS E experiments (sequential acquisition of mass spectra at low and high collision energy) has been crucial to this aim as it allowed promoting fragmentation in the collision cell without any previous precursor ion selection. MDPV belongs to the group of new psychoactive substances (NPS), being known as the "cannibal drug". This substance is being more and more abused and associated with dangerous side effects. The human metabolites (both phase I and phase II) were detected and tentatively identified by accurate-mass full-spectrum measurements using ultra-high performance liquid chromatography coupled to hybrid quadrupole time of flight mass spectrometry (UHPLC-QTOF MS). Following this strategy, up to 10 phase I metabolites, together with some glucuronides and sulphates, were detected and tentative structures were proposed. Several compounds identified in this work have not been previously reported in the literature.
Laboratory wastes containing Cd2+ and water polluted by pharmaceuticals represent an environmenta... more Laboratory wastes containing Cd2+ and water polluted by pharmaceuticals represent an environmental concern. In this work, a proof concept, consisting of the use of teaching-laboratory wastes to synthesize CdS and its subsequent use as a photocatalyst to degrade fluoroquinolone antibiotics, was developed. The CdS was prepared by extraction with thioacetamide and calcination (at 450 °C) and characterized using several techniques. The photocatalytic activity of the CdS, to degrade levofloxacin and norfloxacin, was tested, and the routes involved in the process and the primary transformations of the fluoroquinolones were established. Moreover, the ability of CdS-photocatalysis to eliminate levofloxacin in simulated matrices of fresh urine and hospital wastewater was evaluated. The characterization analyses indicated that the CdS semiconductor was synthesized successfully. Effectively, the CdS acted as a photocatalyst toward degradation of levofloxacin, involving the action of superoxide...
Journal of Environmental Chemical Engineering, 2021
Abstract Discharges from the wastewater treatment plants (WWTPs) are among the main sources of co... more Abstract Discharges from the wastewater treatment plants (WWTPs) are among the main sources of contamination to receiving surface water, therefore the quality of treated wastewater needs to be properly monitored. However, not only the effluents of larger WWTPs employing advanced treatment processes have been considered, but also those from more conventional WWTPs. In this study, the occurrence and behavior of pharmaceuticals have been investigated in a conventional WWTP which receives wastewater from an urban area and a near-by hospital. 24-h composite samples were collected during one week before (influent wastewater, IWW) and after (effluent wastewater, EWW) treatment along three monitoring campaigns distributed over one year. Moreover, seven daily IWW samples discharged from a hospital were also collected. A preliminary wide-scope screening using liquid chromatography (LC) coupled to high resolution mass spectrometry allowed to identify a wide number of pharmaceuticals in the samples. Based on the screening findings, a list of 40 compounds was established for subsequent target quantitative analyses by LC-tandem mass spectrometry. Up to 75% of the compounds investigated were present in all wastewater samples. Analyte concentrations in hospital discharge samples were significantly higher, evidencing an important contribution in terms of pharmaceuticals content. Antibiotics showed the highest concentrations during the winter season, which could be related to the increase in the prescription of these compounds to treat respiratory infections. Data from this work show that the biological treatment applied was able to eliminate nearly half of the compounds under study, although still 12 pharmaceuticals were not or poorly removed.
New psychoactive substances (NPS) have become a serious public health problem, as they are contin... more New psychoactive substances (NPS) have become a serious public health problem, as they are continuously changing their structures, modifying their potency and effects on humans, and therefore novel compounds are unceasingly appearing. One of the major challenges in forensic analysis, particularly related to the problematic of NPS, is the development of fast screening methodologies that allow the detection of a wide variety of compounds in a single analysis. In this study, a novel application of the atmospheric solids analysis probe (ASAP) using medical swabs has been developed. The swab-ASAP was coupled to a triple quadrupole mass analyzer working under data-dependent acquisition mode in order to perform a suspect screening of NPS in different types of samples as well as in surfaces. The compounds were automatically identified based on the observed fragmentation spectra using an in-house built MS/MS spectra library. The developed methodology was applied to the identification of psychoactive substances in research chemicals and herbal blends. The sensitivity of the method, as well as its applicability for surface analysis, was also assessed by identifying down to 1 µg of compound impregnated into a laboratory table. Another remarkable application was the identification of cathinones and synthetic cannabinoids in the fingers of potential consumers. Interestingly, our data showed that NPS could be identified in the fingers after being in contact with the product and even after cleaning their hands by shaking off with a cloth. The methodology proposed in this paper can be applied for routine analyses of NPS in different matrix samples without the need to establish a list of target compounds prior to analysis.
This paper describes a new method for the sensitive and selective determination of fosetyl-alumin... more This paper describes a new method for the sensitive and selective determination of fosetyl-aluminum (Al) residues in vegetable samples. The method involves extraction with water by using a high-speed blender and subsequent injection of the 5-fold diluted extract into the liquid chromatograph. Fosetyl-Al is determined by liquid chromatography with electrospray tandem mass spectrometry after the addition of tetrabutylammonium acetate as the ion-pairing re-agent. The method has been used to assay lettuce samples spiked at 2 and 0.2 mg/kg. Recoveries were satisfactory, with mean values of 98 and 106%, respectively, and relative standard deviations were <10%. The limit of quantitation was 0.2 mg/kg, and the limit of detection was as low as 0.05 mg/kg. Matrix-matched calibration was used for quantitation, and the addition of an internal standard improved repeatability. The developed method allows the accurate and rapid determination of low levels of fosetyl-Al residues in lettuce with ...
The composition of Atlantic salmon feed has changed considerably over the last two decades from b... more The composition of Atlantic salmon feed has changed considerably over the last two decades from being marine-based (fishmeal and fish oil) to mainly containing plant ingredients. Consequently concern related to traditional persistent contaminants typically associated with fish-based feed has been replaced by other potential contaminants not previously associated with salmon farming. This is the case for many pesticides, which are used worldwide to increase food production, and may be present in plant ingredients. Earlier studies have identified two organophosphorus pesticides, chlorpyrifos-methyl and pirimiphos-methyl, in plant ingredients used for aquafeed production. In the present study, we developed a reliable and sensitive analytical method, based on liquid chromatography coupled to tandem mass spectrometry, for the determination of these pesticides and their main metabolites in warm-water (zebrafish) and cold water (Atlantic salmon) species, where possible differences in metabolites could be expected. The method was tested in whole zebrafish and in different salmon tissues, such as muscle, bile, kidney, fat and liver. The final objective of this work was to assess kinetics of chlorpyrifos-methyl and pirimiphos-methyl and their main metabolites in fish tissue, in order to fill the knowledge gaps on these metabolites in fish tissues when fed over prolonged time.
This work compares the sonochemical degradation of losartan and valsartan (antihypertensives) in ... more This work compares the sonochemical degradation of losartan and valsartan (antihypertensives) in water. Initially, the suitable operational conditions of ultrasonic power density and frequency were established. Under such conditions, losartan was eliminated in a higher percentage than valsartan, which was associated to differences in their hydrophobicities. Additionally, degradations in presence of isopropanol and ferrous ions confirmed that losartan was closer to cavitation bubble than valsartan. Afterwards, the structures of primary products indicated that sonogenerated hydroxyl radical attacked biphenyl tetrazole moiety (common nucleus of both pharmaceuticals). Then, theoretical calculations were applied to the products to estimate the toxicity, degree of oxidation and probable routes of aerobic biodegradation suggesting a beneficial action of sonodegradation. Finally, the sonochemical degradation of the antihypertensives was carried out in two simulated complex matrices (i.e., seawater and hospital wastewater) and an actual wastewater. Interestingly, the losartan and valsartan eliminations in such waters was similar to the observed in distilled water. This fact indicates the high potentiality of ultrasound to degrade losartan or valsartan in waters containing other substances, even at higher concentrations than these pollutants.
Despite the fact that 33% of the new psychoactive substances seized in 2015 were synthetic cathin... more Despite the fact that 33% of the new psychoactive substances seized in 2015 were synthetic cathinones, the number of these derivatives has been decreasing in the last years, probably as a consequence of the unfavourable effects reported by users. Thus, the list of possible cathinone analogues is expected to get shorter, and it is likely that the same moiety changes applied for the preparation of synthetic cathinones will be applied in the near future to other stimulants in the search for favourable alternatives to controlled substances. This is evidenced by the increase in newly reported substances belonging to stimulant classes other than cathinones. One of the possible candidates for a new backbone from which to base new stimulants is aminorex, which is classified as a Schedule I substance by the Drug Enforcement Administration. Three derivatives have been reported until now: 4-methylaminorex or 4-MAR (also categorized as a Schedule I substance), para-methyl-4-methylaminorex (4,4′-DMAR) and 3′,4′-methylenedioxy-4-methylaminorex (MDMAR). Recently, the new halogenated 4-MAR derivative, para-fluoro-4-methylaminorex, characterised in this work (and abbreviated as pF-4-methylaminorex or 4′F-4-MAR) was detected by the Slovenian police. In the present work, 4′F-4-MAR has been characterized by high resolution mass spectrometry and nuclear magnetic resonance in a sample obtained from an anonymous consumer. This research shows that the same modifications applied for the preparation of synthetic cathinones are being used to prepare new stimulants based on the aminorex backbone.
New psychoactive substances have been rapidly growing in popularity in the drug market as non-ill... more New psychoactive substances have been rapidly growing in popularity in the drug market as non-illegal drugs. In the last few years, an increment has been reported on the use of synthetic alternatives to heroin, the synthetic opioids. Based on the information provided by the European Monitoring Centre for Drug and Drug Addiction, these synthetic opioids have been related to overdoses and deaths in Europe and North America. One of these opioids is the U-47700. A few months ago, U-47700 was scheduled in the U.S. and other countries, and other opioid derivatives have been appearing in order to replace it. One of these compounds is U-49900, an analog of U-47700. A white powder sample was obtained from an anonymous user in Spain. After an accurate characterization by gas chromatography-mass spectrometry, ultra-high performance liquid chromatography-high resolution mass spectrometry, nuclear magnetic resonance and single-crystal X-ray diffraction; and complemented by Fouriertransformed infrared spectroscopy, ultraviolet and circular dichroism spectrophotometry, the drug sample was unequivocally identified as U-49900. The information provided will be useful for the Early Warning System and forensic laboratories for future identifications of the U-49900, as well as in tentative identifications of other related opioids.
Chemical analysis of raw wastewater in order to assess the presence of biological markers enterin... more Chemical analysis of raw wastewater in order to assess the presence of biological markers entering a wastewater treatment plant can provide objective information about the health and lifestyle of the population connected to the sewer system. This work was performed in a tropical country of Central America, Costa Rica, with the aim of extending this knowledge to new world regions. This work is the first to report wastewater-based epidemiological data on the use of illicit drugs in this region of the world. Composite wastewater samples from the influents of two different wastewater treatment facilities and surface water samples from surrounding areas were collected applying the best practice protocol and analysed to investigate the occurrence and fate of selected illicit drugs of abuse and pharmaceuticals. Results showed the presence of chemical indicators of the classic drugs cocaine and cannabis at high concentration levels, besides the moderate presence of the opiates codeine and m...
Wastewater-based epidemiology (WBE) employs the analysis of wastewater to detect and quantify dru... more Wastewater-based epidemiology (WBE) employs the analysis of wastewater to detect and quantify drug use and discharge within a community. In this work, transformation products (TP) by microbes in the environment were identified after incubations in wastewater and an isolated microbial strain. The microbial strain was isolated from an enrichment culture of wastewater supplemented with 3,4-methylenedioxy-pyrovalerone, and identified by matrix assisted laser desorption - time of flight mass spectrometry as Pseudomonas putida (P. putida). Five pyrrolidinophenone-type psychoactive substances (PPPS) were then incubated in wastewater and in P. putida tryptic soy broth (TSB) growth cultures. TPs were identified using liquid chromatography coupled to mass spectrometry techniques. All TPs observed in P. putida TSB growth cultures were also identified in wastewater incubations. The main TP for all PPPSs in P. putida TSB growth cultures, and two PPPSs incubated in wastewater, were the N-desalkyl...
This paper provides scientific results from a European LIFE project carried out in the Valencian ... more This paper provides scientific results from a European LIFE project carried out in the Valencian region of Spain during the 2017 to 2018 time frame. In 2018, more than 60,000 tons of pesticides were commercialized in Spain, with approximately 15% destined for Valencian crops. In order to improve the air quality in the agricultural areas of this region, an innovative cropping system based on irrigation was developed and compared to conventional treatments based on hand-spray and turbo application. After applying conventional treatments to five types of crops (citrus, persimmon, nectarine, watermelon, and other stone fruits), a total of 13 active substances were detected in the air. The same active substances were applied to crops using the novel irrigation system, and no pesticide was detected in the air. Moreover, applicator and bystander populations in the region were assessed for their risk of inhalation exposure to pesticides, and no risk was found when either of the techniques, ...
The interest of ambient ionization mass spectrometry in clinical and forensic analysis has increa... more The interest of ambient ionization mass spectrometry in clinical and forensic analysis has increased in the last years as it allows the rapid and direct analysis of a wide variety of samples. Among the possible applications, the analysis in real-time of the exhaled breath has gained great attention, for the investigation of both endogenous compounds, as for example disease biomarkers, and exogenous compounds related to the consumption of certain products. Although commercial ionization sources are already available for breath analysis, they require an important economic investment and/or complex setups. In this article, we describe a new probe developed at our laboratory, fully compatible with the Waters Corp. atmospheric solids analysis probe (ASAP), and where manufacturing is neither expensive nor complex. This atmospheric breath analysis probe (ABAP) can be directly used in Waters mass spectrometry instruments without the need of modifying the ionization source, the instrument or the ASAP, being also compatible with the software provided by the manufacturer. The ionization of the compounds is based on atmospheric pressure chemical ionization. The prototype has been successfully used in a high-resolution mass spectrometry system, and applied to the analysis of compounds present in human breath, as well as to the tentative identification of different substances present in foods and reported in exhaled breath after food consumption. In addition, ABAP could also be compatible with other ionization techniques, such as photoionization.
Currently, the most powerful approach to monitor organic micropollutants (OMPs) in environmental ... more Currently, the most powerful approach to monitor organic micropollutants (OMPs) in environmental samples is the combination of target, suspect and non-target screening strategies using high resolution mass spectrometry (HRMS). However, the high complexity of sample matrices as well as the huge number of OMPs potentially present in samples at low concentrations pose an analytical challenge. Ion mobility separation (IMS) combined with HRMS instruments (IMS-HRMS) introduces an additional analytical dimension, providing extra information which facilitates the identification of OMPs. The collision cross section (CCS) value provided by IMS is unaffected by the matrix or chromatographic separation. Consequently, the creation of CCS databases and the inclusion of ion mobility within identification criteria are of high interest for an enhanced and robust screening strategy. In this work, a CCS library for IMS-HRMS, which is online and freely available, was developed for 556 OMPs in both positive and negative ionization modes using electrospray ionization. The inclusion of ion mobility data in widely adopted confidence levels for identification in environmental reporting is discussed. Illustrative examples of OMPs found in environmental samples are presented to highlight the potential of IMS-HRMS and to demonstrate the additional value of CCS data in various screening strategies.
The pesticide chlorpyrifos-methyl (CLP-m) has been identified in plant ingredients intended for a... more The pesticide chlorpyrifos-methyl (CLP-m) has been identified in plant ingredients intended for aquaculture feed production. To investigate systemic effects of CLP-m with emphasis on lipid metabolism, post-smolt Atlantic salmon were fed in duplicate (n=2) either diets with no CLP-m (Control) or CLP-m at different concentrations (0.1, 1.0 or 8.0 mg kg-1) for a total of 67 days (Low, Medium, High). Fish in all groups almost doubled their weight during the feeding trial from 262 ± 26g (mean ± SD) to 465 ± 64g (overall mean), with no significant effects on any growth parameters. There was a significant dosedependent inhibition of plasma cholinesterase activity (BuChE) after 67 days. The CLP-m biotransformation metabolite, TCP was detected in liver and bile, with low levels of the parent compound in the organs. Spleen somatic index decreased significantly with increasing dietary CLP-m intake. Hematocrit (%) decreased linearly with increasing dietary exposure to CLP-m after 30 days of exposure, but this decrease was less at 67 days of exposure. A significantly reduced content of arachidonic acid (ARA 20:4n-6), accompanied by a significantly increased content of the saturated fatty acid, palmitic acid (PA 16:0), was observed in liver phospholipids (PLs) with increasing dietary content of CLP-m. Major effects were seen on the PL classes in liver which showed a significantly decreased absolute content, possibly indicating inhibition of PL remodeling pathways or other membrane perturbation effects from CLP-m exposure. In conclusion, this study shows that the pesticide CLP-m is a relatively potent toxicant in Atlantic salmon, especially affecting liver PLs and ARA metabolism.
The potential of ultra-high-pressure liquid chromatography (UHPLC) coupled to hybrid quadrupole t... more The potential of ultra-high-pressure liquid chromatography (UHPLC) coupled to hybrid quadrupole timeof-flight mass spectrometry (QTOF-MS) for the screening and confirmation of antibiotics in water samples is illustrated in this paper. UHPLC presents several advantages over conventional liquid chromatography as it generates narrow peaks (increasing peak height and improving sensitivity) and reduces chromatographic runs. Regarding QTOF-MS, its increased mass resolution, high sensitivity in full-spectrum acquisition mode and mass accuracy, in both MS and MS/MS modes, make this technique ideal for the detection and reliable confirmation of organic contaminants in environmental samples. After a solidphase extraction using Oasis HLB cartridges, UHPLC-QTOF-MS has been applied in this work to several types of water samples (surface water and influent and effluent wastewaters). Several antibiotics were found in the samples, such as ofloxacin, ciprofloxacin, clarythromycin or erythromycin, among others. Moreover, the full spectrum data provided by TOF-MS acquisition has enabled searching for many other pharmaceuticals that could be present in the samples in a "post-target" way. This approach has allowed the detection and confirmation of paracetamol, omeprazole and codeine, among others. UPLC-QTOF-MS has been shown as an attractive and efficient hyphenated technique for the rapid detection and confirmation of pharmaceuticals in water with very little sample handling.
A fast and automated strategy has been developed for identifying unknown substances in the atmosp... more A fast and automated strategy has been developed for identifying unknown substances in the atmosphere (concretely, in the particulate matter, PM10) using LC-HRMS (MS3). A total of 15 samples were collected in three different areas (rural, urban and industrial). A sampling flow rate of 30 m3 h−1 was applied for 24 h, sampling a total volume of around 720 m3. A total of 49 compounds were tentatively identified using very restrictive criteria regarding exact mass, retention time, isotopic profile and both MS2 and MS3 spectra. Pesticides, pharmaceutical active compounds, drugs, plasticizers and metabolites were the most identified compounds. To verify whether the developed methodology was suitable, 11 substances were checked with their analytical standards and all of them were confirmed. Different profiles for industrial, rural and urban areas were examined. The Principal Component Analysis (PCA) model allowed us to separate the obtained data of the three assessed area. When the profile...
Synthetic cannabinoids receptor agonists (SCRAs) are often almost completely metabolised, and hen... more Synthetic cannabinoids receptor agonists (SCRAs) are often almost completely metabolised, and hence their pharmacokinetics should be carefully evaluated for determining the most adequate biomarker in toxicological analysis. Two structurally related SCRAs, AMB-FUBINACA and AMB-CHMICA, were selected to evaluate their in vivo metabolism and pharmacokinetics using male Sprague-Dawley rats. Brain, liver, kidney, blood (serum) and urine samples were collected at different times to assess the differences in metabolism, metabolic reactions, tissue distribution and excretion. Both compounds experimented O-demethyl reaction, which occurred more rapidly for AMB-FUBINACA. The parent compounds and O-demethyl metabolites were highly bioaccumulated in liver, and were still detected in this tissue 48 h after injection. The different indazole/indole N-functionalisation produced diverse metabolic reactions in this moiety and thus, different urinary metabolites were formed. Out of the two compounds, A...
The potential of high resolution mass spectrometry (HRMS) for the investigation of human in vivo ... more The potential of high resolution mass spectrometry (HRMS) for the investigation of human in vivo metabolism of 3,4-methylenedioxypyrovalerone (MDPV) using urine collected from a consumer (this is, in non-controlled experiments) has been investigated. As a control sample was not available, the common approach based on the comparison of a control/blank sample and samples collected after drug intake could not be used. Alternatively, an investigation based on common fragmentation pathways was applied, assuming that most metabolites share some fragments with the parent drug. An extension of this approach was also applied based on the fragmentation pathway of those metabolites identified in urine samples in the first step. The use of MS E experiments (sequential acquisition of mass spectra at low and high collision energy) has been crucial to this aim as it allowed promoting fragmentation in the collision cell without any previous precursor ion selection. MDPV belongs to the group of new psychoactive substances (NPS), being known as the "cannibal drug". This substance is being more and more abused and associated with dangerous side effects. The human metabolites (both phase I and phase II) were detected and tentatively identified by accurate-mass full-spectrum measurements using ultra-high performance liquid chromatography coupled to hybrid quadrupole time of flight mass spectrometry (UHPLC-QTOF MS). Following this strategy, up to 10 phase I metabolites, together with some glucuronides and sulphates, were detected and tentative structures were proposed. Several compounds identified in this work have not been previously reported in the literature.
Laboratory wastes containing Cd2+ and water polluted by pharmaceuticals represent an environmenta... more Laboratory wastes containing Cd2+ and water polluted by pharmaceuticals represent an environmental concern. In this work, a proof concept, consisting of the use of teaching-laboratory wastes to synthesize CdS and its subsequent use as a photocatalyst to degrade fluoroquinolone antibiotics, was developed. The CdS was prepared by extraction with thioacetamide and calcination (at 450 °C) and characterized using several techniques. The photocatalytic activity of the CdS, to degrade levofloxacin and norfloxacin, was tested, and the routes involved in the process and the primary transformations of the fluoroquinolones were established. Moreover, the ability of CdS-photocatalysis to eliminate levofloxacin in simulated matrices of fresh urine and hospital wastewater was evaluated. The characterization analyses indicated that the CdS semiconductor was synthesized successfully. Effectively, the CdS acted as a photocatalyst toward degradation of levofloxacin, involving the action of superoxide...
Journal of Environmental Chemical Engineering, 2021
Abstract Discharges from the wastewater treatment plants (WWTPs) are among the main sources of co... more Abstract Discharges from the wastewater treatment plants (WWTPs) are among the main sources of contamination to receiving surface water, therefore the quality of treated wastewater needs to be properly monitored. However, not only the effluents of larger WWTPs employing advanced treatment processes have been considered, but also those from more conventional WWTPs. In this study, the occurrence and behavior of pharmaceuticals have been investigated in a conventional WWTP which receives wastewater from an urban area and a near-by hospital. 24-h composite samples were collected during one week before (influent wastewater, IWW) and after (effluent wastewater, EWW) treatment along three monitoring campaigns distributed over one year. Moreover, seven daily IWW samples discharged from a hospital were also collected. A preliminary wide-scope screening using liquid chromatography (LC) coupled to high resolution mass spectrometry allowed to identify a wide number of pharmaceuticals in the samples. Based on the screening findings, a list of 40 compounds was established for subsequent target quantitative analyses by LC-tandem mass spectrometry. Up to 75% of the compounds investigated were present in all wastewater samples. Analyte concentrations in hospital discharge samples were significantly higher, evidencing an important contribution in terms of pharmaceuticals content. Antibiotics showed the highest concentrations during the winter season, which could be related to the increase in the prescription of these compounds to treat respiratory infections. Data from this work show that the biological treatment applied was able to eliminate nearly half of the compounds under study, although still 12 pharmaceuticals were not or poorly removed.
New psychoactive substances (NPS) have become a serious public health problem, as they are contin... more New psychoactive substances (NPS) have become a serious public health problem, as they are continuously changing their structures, modifying their potency and effects on humans, and therefore novel compounds are unceasingly appearing. One of the major challenges in forensic analysis, particularly related to the problematic of NPS, is the development of fast screening methodologies that allow the detection of a wide variety of compounds in a single analysis. In this study, a novel application of the atmospheric solids analysis probe (ASAP) using medical swabs has been developed. The swab-ASAP was coupled to a triple quadrupole mass analyzer working under data-dependent acquisition mode in order to perform a suspect screening of NPS in different types of samples as well as in surfaces. The compounds were automatically identified based on the observed fragmentation spectra using an in-house built MS/MS spectra library. The developed methodology was applied to the identification of psychoactive substances in research chemicals and herbal blends. The sensitivity of the method, as well as its applicability for surface analysis, was also assessed by identifying down to 1 µg of compound impregnated into a laboratory table. Another remarkable application was the identification of cathinones and synthetic cannabinoids in the fingers of potential consumers. Interestingly, our data showed that NPS could be identified in the fingers after being in contact with the product and even after cleaning their hands by shaking off with a cloth. The methodology proposed in this paper can be applied for routine analyses of NPS in different matrix samples without the need to establish a list of target compounds prior to analysis.
This paper describes a new method for the sensitive and selective determination of fosetyl-alumin... more This paper describes a new method for the sensitive and selective determination of fosetyl-aluminum (Al) residues in vegetable samples. The method involves extraction with water by using a high-speed blender and subsequent injection of the 5-fold diluted extract into the liquid chromatograph. Fosetyl-Al is determined by liquid chromatography with electrospray tandem mass spectrometry after the addition of tetrabutylammonium acetate as the ion-pairing re-agent. The method has been used to assay lettuce samples spiked at 2 and 0.2 mg/kg. Recoveries were satisfactory, with mean values of 98 and 106%, respectively, and relative standard deviations were <10%. The limit of quantitation was 0.2 mg/kg, and the limit of detection was as low as 0.05 mg/kg. Matrix-matched calibration was used for quantitation, and the addition of an internal standard improved repeatability. The developed method allows the accurate and rapid determination of low levels of fosetyl-Al residues in lettuce with ...
The composition of Atlantic salmon feed has changed considerably over the last two decades from b... more The composition of Atlantic salmon feed has changed considerably over the last two decades from being marine-based (fishmeal and fish oil) to mainly containing plant ingredients. Consequently concern related to traditional persistent contaminants typically associated with fish-based feed has been replaced by other potential contaminants not previously associated with salmon farming. This is the case for many pesticides, which are used worldwide to increase food production, and may be present in plant ingredients. Earlier studies have identified two organophosphorus pesticides, chlorpyrifos-methyl and pirimiphos-methyl, in plant ingredients used for aquafeed production. In the present study, we developed a reliable and sensitive analytical method, based on liquid chromatography coupled to tandem mass spectrometry, for the determination of these pesticides and their main metabolites in warm-water (zebrafish) and cold water (Atlantic salmon) species, where possible differences in metabolites could be expected. The method was tested in whole zebrafish and in different salmon tissues, such as muscle, bile, kidney, fat and liver. The final objective of this work was to assess kinetics of chlorpyrifos-methyl and pirimiphos-methyl and their main metabolites in fish tissue, in order to fill the knowledge gaps on these metabolites in fish tissues when fed over prolonged time.
This work compares the sonochemical degradation of losartan and valsartan (antihypertensives) in ... more This work compares the sonochemical degradation of losartan and valsartan (antihypertensives) in water. Initially, the suitable operational conditions of ultrasonic power density and frequency were established. Under such conditions, losartan was eliminated in a higher percentage than valsartan, which was associated to differences in their hydrophobicities. Additionally, degradations in presence of isopropanol and ferrous ions confirmed that losartan was closer to cavitation bubble than valsartan. Afterwards, the structures of primary products indicated that sonogenerated hydroxyl radical attacked biphenyl tetrazole moiety (common nucleus of both pharmaceuticals). Then, theoretical calculations were applied to the products to estimate the toxicity, degree of oxidation and probable routes of aerobic biodegradation suggesting a beneficial action of sonodegradation. Finally, the sonochemical degradation of the antihypertensives was carried out in two simulated complex matrices (i.e., seawater and hospital wastewater) and an actual wastewater. Interestingly, the losartan and valsartan eliminations in such waters was similar to the observed in distilled water. This fact indicates the high potentiality of ultrasound to degrade losartan or valsartan in waters containing other substances, even at higher concentrations than these pollutants.
Despite the fact that 33% of the new psychoactive substances seized in 2015 were synthetic cathin... more Despite the fact that 33% of the new psychoactive substances seized in 2015 were synthetic cathinones, the number of these derivatives has been decreasing in the last years, probably as a consequence of the unfavourable effects reported by users. Thus, the list of possible cathinone analogues is expected to get shorter, and it is likely that the same moiety changes applied for the preparation of synthetic cathinones will be applied in the near future to other stimulants in the search for favourable alternatives to controlled substances. This is evidenced by the increase in newly reported substances belonging to stimulant classes other than cathinones. One of the possible candidates for a new backbone from which to base new stimulants is aminorex, which is classified as a Schedule I substance by the Drug Enforcement Administration. Three derivatives have been reported until now: 4-methylaminorex or 4-MAR (also categorized as a Schedule I substance), para-methyl-4-methylaminorex (4,4′-DMAR) and 3′,4′-methylenedioxy-4-methylaminorex (MDMAR). Recently, the new halogenated 4-MAR derivative, para-fluoro-4-methylaminorex, characterised in this work (and abbreviated as pF-4-methylaminorex or 4′F-4-MAR) was detected by the Slovenian police. In the present work, 4′F-4-MAR has been characterized by high resolution mass spectrometry and nuclear magnetic resonance in a sample obtained from an anonymous consumer. This research shows that the same modifications applied for the preparation of synthetic cathinones are being used to prepare new stimulants based on the aminorex backbone.
New psychoactive substances have been rapidly growing in popularity in the drug market as non-ill... more New psychoactive substances have been rapidly growing in popularity in the drug market as non-illegal drugs. In the last few years, an increment has been reported on the use of synthetic alternatives to heroin, the synthetic opioids. Based on the information provided by the European Monitoring Centre for Drug and Drug Addiction, these synthetic opioids have been related to overdoses and deaths in Europe and North America. One of these opioids is the U-47700. A few months ago, U-47700 was scheduled in the U.S. and other countries, and other opioid derivatives have been appearing in order to replace it. One of these compounds is U-49900, an analog of U-47700. A white powder sample was obtained from an anonymous user in Spain. After an accurate characterization by gas chromatography-mass spectrometry, ultra-high performance liquid chromatography-high resolution mass spectrometry, nuclear magnetic resonance and single-crystal X-ray diffraction; and complemented by Fouriertransformed infrared spectroscopy, ultraviolet and circular dichroism spectrophotometry, the drug sample was unequivocally identified as U-49900. The information provided will be useful for the Early Warning System and forensic laboratories for future identifications of the U-49900, as well as in tentative identifications of other related opioids.
Chemical analysis of raw wastewater in order to assess the presence of biological markers enterin... more Chemical analysis of raw wastewater in order to assess the presence of biological markers entering a wastewater treatment plant can provide objective information about the health and lifestyle of the population connected to the sewer system. This work was performed in a tropical country of Central America, Costa Rica, with the aim of extending this knowledge to new world regions. This work is the first to report wastewater-based epidemiological data on the use of illicit drugs in this region of the world. Composite wastewater samples from the influents of two different wastewater treatment facilities and surface water samples from surrounding areas were collected applying the best practice protocol and analysed to investigate the occurrence and fate of selected illicit drugs of abuse and pharmaceuticals. Results showed the presence of chemical indicators of the classic drugs cocaine and cannabis at high concentration levels, besides the moderate presence of the opiates codeine and m...
Wastewater-based epidemiology (WBE) employs the analysis of wastewater to detect and quantify dru... more Wastewater-based epidemiology (WBE) employs the analysis of wastewater to detect and quantify drug use and discharge within a community. In this work, transformation products (TP) by microbes in the environment were identified after incubations in wastewater and an isolated microbial strain. The microbial strain was isolated from an enrichment culture of wastewater supplemented with 3,4-methylenedioxy-pyrovalerone, and identified by matrix assisted laser desorption - time of flight mass spectrometry as Pseudomonas putida (P. putida). Five pyrrolidinophenone-type psychoactive substances (PPPS) were then incubated in wastewater and in P. putida tryptic soy broth (TSB) growth cultures. TPs were identified using liquid chromatography coupled to mass spectrometry techniques. All TPs observed in P. putida TSB growth cultures were also identified in wastewater incubations. The main TP for all PPPSs in P. putida TSB growth cultures, and two PPPSs incubated in wastewater, were the N-desalkyl...
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Papers by María Ibáñez