Identification of ethoxyquin and its transformation products in salmon after controlled dietary exposure via fish feed

Ethoxyquin (EQ) is an additive present in fish feed and its fate in fish should be carefully characterized due to food safety concerns regarding this compound. Therefore, the objective of this work was to identify the transformation products (TPs) of EQ in Atlantic salmon. Salmon in independent tanks were given feed containing ethoxyquin concentrations of 0.5 mg/kg, 119 mg/kg or 1173 mg/kg for 90 days. After the feeding trial, salmon fillets were extracted in acetonitrile and analyzed by liquid chromatography with traveling-wave ion mobility spectrometry coupled to high resolution mass spectrometry (UHPLC-TWIMS-QTOFMS). EQ was transferred from the feed to salmon fillets and 23 TPs were characterized, resulting from dimerization, oxygenation, cleavage, cleavage combined with oxygenation, cleavage combined with conjugation, and other uncategorized alterations. Moreover, EQ and some TPs were also detected in commercial salmon randomly sampled from different Norwegian fish farms. This study confirmed that the dimer 1,8′-EQDM was the main TP of EQ and, together with previous research, brought the overall number of characterized TPs to a total of 47.publishedVersio

A data-independent acquisition workflow for qualitative screening of new psychoactive substances in biological samples

Identification of new psychoactive substances (NPS) is challenging. Developing targeted methods for their analysis can be difficult and costly due to their impermanence on the drug scene. Accurate-mass mass spectrometry (AMMS) using a quadrupole time-of-flight (QTOF) analyzer can be useful for wide-scope screening since it provides sensitive, full-spectrum MS data. Our article presents a qualitative screening workflow based on data-independent acquisition mode (all-ions MS/MS) on liquid chromatography (LC) coupled to QTOFMS for the detection and identification of NPS in biological matrices. The workflow combines and structures fundamentals of target and suspect screening data processing techniques in a structured algorithm. This allows the detection and tentative identification of NPS and their metabolites. We have applied the workflow to two actual case studies involving drug intoxications where we detected and confirmed the parent compounds ketamine, 25B-NBOMe, 25C-NBOMe, and several predicted phase I and II metabolites not previously reported in urine and serum samples. The screening workflow demonstrates the added value for the detection and identification of NPS in biological matrices

On-line solid phase extraction-liquid chromatography-tandem mass spectrometry for the determination of 17 cytostatics and metabolites in waste, surface and ground water samples

A fully automated on-line solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) method has been developed for the determination of 13 cytostatics and 4 metabolites in aqueous matrices, including groundwater, surface water, and raw and treated wastewater. On-line SPE is performed by loading 5mL of water sample at pH 2 through a PLRP-s cartridge. MS/MS is performed with an electrospray (ESI) interface operating in the positive ion mode and registering two selected reaction monitoring (SRM) transitions per compound. Quantification is carried out by the isotope dilution method using 15 different isotope-labelled compounds, specific for the target analytes, as internal standards (IS). The main advantages of the method are high sensitivity, with limits of determination in groundwater, surface water, and raw and treated wastewater below 5ngL-1 for all compounds except for gemcitabine (6.9-9.3ngL-1), temozolomide (26-50ngL-1), imatinib (80-180ngL-1) and etoposide (38-65ngL-1), repeatability, with relative standard deviations (RSDs) in most cases below 15%, and selectivity and reliability of results. The method is also fairly simple and fast, with an analysis time per sample (excluding the manual steps, i.e., sample filtration, pH adjustment, and addition of IS) of 40min. Application of the method to influent wastewater samples collected daily during eight consecutive days from a wastewater treatment plant (WWTP) from Catalonia showed the presence of methotrexate, ifosfamide, capecitabine, tamoxifen and 6(α)-hydroxypaclitaxel but at fairly low concentrations (up to 43ngL-1). © 2013 Elsevier B.V.This study was financially supported by the EU through the EU FP7 project CytoThreat (Fate and effects of cytostatic pharmaceuticals in the environment and the identification of biomarkers for and improved risk assessment on environmental exposure. Grant agreement No.: 265264), by the Spanish Ministry of Economy and Competitiveness through the projects SCARCE (Consolider-Ingenio 2010 CSD2009-00065) and CEMAGUA (CGL2007-64551/HID), and by the Generalitat de Catalunya (Consolidated Research Group: Water and Soil Quality Unit 2009-SGR-965). It reflects only the author's views. The Community is not liable for any use that may be made of the information contained therein. Merck and Biotage are acknowledged for the gift of LC columns and SPE cartridges, respectively. Damià Barceló acknowledges financial support from the Visiting Professor Programme of the King Saud University, Riyadh, Saudi Arabia. Pedro Aguiló, Joaquín Bosque and Begoña Martínez from EMSSA (Barcelona, Spain) are acknowledged for their collaboration in the sampling of wastewaters.Peer Reviewe

Multianalyte determination of 24 cytostatics and metabolites by liquid chromatography-electrospray-tandem mass spectrometry and study of their stability and optimum storage conditions in aqueous solution

A multianalyte liquid chromatography-electrospray-tandem mass spectrometry (LC-ESI-MS/MS) method for determination of 19 cytostatics and 5 metabolites, from 6 different therapeutic families, has been developed, and the structures of the main characteristic fragment ions have been proposed. Instrumental limits of detection and quantification are in the range 0.1-10.3 and 1.0-34.3 ng mL−1, respectively. Moreover, the stability of the compounds in aqueous solution was investigated in order to establish the best conditions for preparation and storage of both calibration standards and water samples. Dimethylsulfoxide (DMSO) was selected as solvent for preparation of the stock solutions. At room temperature (25 ºC), 11 of the 24 target compounds were shown to be unstable in water (percentage of organic solvent 4%), with concentration losses greater than 20% in less than 24 h. At 4 ºC (typical storage temperature for water samples) all compounds, except MTIC and chlorambucil, were stable for 24 h, but the number of stable compounds decreased to 10 after 9 days. Freezing of the aqueous solutions improved considerably the stability of various compounds: after 3 months of storage at -20 ºC, 10 compounds, namely, 5-fluorouracil, carboplatin, gemcitabine, temozolomide, vincristine, vinorelbine, ifosfamide, cyclophosphamide, etoposide, and capecitabine, remained stable (in contrast to only carboplatin and capecitabine at 4 ºC). The addition of acid improved the stability of methotrexate and its metabolite hydroxymethotrexate but not that of the rest of compounds. The addition of organic solvent (50% methanol or DMSO) prevented the degradation at 4 ºC of the otherwise unstable compounds oxaliplatin, methotrexate, erlotinib, doxorubicin, tamoxifen, and paclitaxel. To the authors' knowledge, five of the analytes investigated have never been searched for in the aquatic environment (imatinib, 6α-hydroxypaclitaxel, endoxifen, (Z)4- Hydroxytamoxifen, and temozolomide), and for many of them the stability data provided, and even the analytical LC-MS/MS conditions, are the first ever published.This study was financially supported by the EU through the EU FP7 project CytoThreat(Fate and effects of cytostatic pharmaceuticals in the environment and the identification of biomarkers for and improved risk assessment on environmental exposure. Grant agreement No.: 265264), by the Spanish Ministry of Economy and Competitiveness through the projects SCARCE (Consolider-Ingenio 2010 CSD2009-00065) and CEMAGUA (CGL2007-64551/HID), and by the Generalitat de Catalunya (Consolidated Research Group: Water and Soil Quality Unit 2009-SGR-965). Nicola Mastroianni acknowledges his CSIC-JAE predoctoral fellowship. It reflects only the author’s views. The Community is not liable for any use that may be made of the information contained therein. Merck is acknowledged for the gift of LC columns, respectively. Damià Barceló acknowledges financial support from the Visiting Professor Program of the King Saud University, Riyadh, Saudi Arabia.Peer reviewe

Aerobic activated sludge transformation of methotrexate: Identification of biotransformation products

This study describes the biotransformation of cytostatic and immunosuppressive pharmaceutical methotrexate. Its susceptibility to microbiological breakdown was studied in a batch biotransformation system, in presence or absence of carbon source and at two activated sludge concentrations. The primary focus of the present study are methotrexate biotransformation products, which were tentatively identified by the ultra-high performance liquid chromatography-quadrupole - Orbitrap-MS. Data-dependent experiments, combining full-scan MS data with product ion spectra were acquired, in order to identify the molecular ions of methotrexate transformation products, to propose the molecular formulae and to elucidate their chemical structures. Among the identified transformation products 2,4-diamino-N10-methyl-pteroic acid is most abundant and persistent. Other biotransformation reactions involve demethylation, oxidative cleavage of amine, cleavage of C-N bond, aldehyde to carboxylate transformation and hydroxylation. Finally, a breakdown pathway is proposed, which shows that most of methotrexate breakdown products retain the diaminopteridine structural segment. In total we propose nine transformation products, among them eight are described as methotrexate transformation products for the first time.a Jožef Stefan Institute, Department of Environmental Sciences, Jamova 39, Ljubljana, Slovenia b Water and Soil Quality Research Group, Department of Environmental Chemistry, Institute of Environmental Assessment and Water Research (IDAEA-CSIC), Jordi Girona 18-26, Barcelona, Spain c Catalan Institute for Water Research (ICRA), Scientific and Technological Park of the University of Girona, H2O Building Emili Grahit 101, Girona, SpainPeer reviewe

A novel workflow utilizing open-source software tools in the environmental fate studies: The example of imatinib biotransformation

The aim of this study is to utilize novel and powerful workflows with publicly available tools to efficiently process data and facilitate rapid acquisition of knowledge on environmental fate studies. Taking imatinib (IMA) as an example, we developed an efficient workflow to describe IMA biodegradation with activated sludge (AS) from wastewater treatment plants (WWTP). IMA is a cytostatic pharmaceutical; a selective tyrosine kinase inhibitor used to treat chronic myeloid leukemia. Its reported ecotoxic, endocrine and genotoxic effects imply high risk for aquatic wildlife and human health, however its fate in the environment is not yet well known. The study was conducted in a batch biotransformation setup, at two AS concentration levels and in presence and absence of carbon source. Degradation profiles and formation of IMA transformation products (TPs) were investigated using UHPLC-QqOrbitrap-MS/MS which showed that IMA is readily biodegradable. TPs were determined using multivariate statistical analysis. Eight TPs were determined and tentatively identified, six of them for first time. Hydrolysis of amide bond, oxidation, demethylation, deamination, acetylation and succinylation are proposed as major biodegradation pathways. TP235, the product of amide bond hydrolysis, was detected and quantified in actual wastewaters, at levels around 1 ng/L. This calls for more studies on the environmental fate of IMA in order to properly asses the environmental risk and hazard associated to IMA and its TPs.This work has received funding from the European Commission through the projects CytoThreat (265264) and SOLUTIONS (603437), from Slovenian Research Agency - ARRS program P1-0143: Cycling of substances in the environment, mass balances, modelling of environmental processes and risk assessment, ARRS projects J1-8140 (Endocrine disrupting and genotoxic potential of protein kinase inhibitors: relevance for environmental hazard and human health) and J1-6744 (Development of Molecularly Imprinted Polymers and their application in environmental and bio-analysis), the Spanish Ministry of Science and Innovation (Project CEX2018-000794-S), and the Generalitat de Catalunya (Consolidated Research Group 2017 SGR 01404-Water and Soil Quality Unit). Žiga Tkalec acknowledges the Slovenian Research Agency – ARRS for the Young researcher grant. The work reflects only the authors' views. The Community in not liable for any use that may be made of the information contained therein. The authors thank Vesna Mislej from Central Wastewater Treatment Plant in Ljubljana and Marjetka Levstek from Wastewater treatment plant Domžale-Kamnik for providing wastewater samples. Special thanks to Metka Filipič from the National institute of biology for providing the imatinib consumption data.Peer reviewe

UHPLC-HRMS data from non-targeted screening for biotransformation products of cytostatic drug imatinib

Imatinib is a selective tyrosine kinase inhibitor used to treat chronic myeloid leukemia. It enters the environment by excretion from the body through urine and feces and is transferred with wastewater to a wastewater treatment plant. There, it can be degraded by activated sludge, forming a number of biotransformation products. Presence of imatinib and its potential transformation products in the environment can impose a high risk to aquatic organisms and human health, therefore it is important to obtain knowledge of its environmental fate. The data presented here is a result of a simulated biodegradation of imatinib at two levels of activated sludge using a batch biotransformation setup, with and without carbon source. The data was acquired with UHPLC-HRMS/MS and processed by MzMine2.36 [1]. The dataset presents a table of [M+H]+ features with retention times and corresponding MS/MS data. With development of new data mining tools this data can be used to identify new transformation products of imatinib and with it fully understand its environmental fate and the risk associated with its presence in the environment.This work has received funding from the European Commission through the projects CytoThreat (265264) and SOLUTIONS (603437), the Slovenian Research Agency - ARRS program P1-0143: Cycling of substances in the environment, mass balances, modelling of environmental processes and risk assessment the Spanish Ministry of Science and Innovation (Project CEX2018-000794-S), and the Generalitat de Catalunya (Consolidated Research Group 2017 SGR 01404-Water and Soil Quality Unit). Žiga Tkalec acknowledges the Slovenian Research Agency – ARRS for the Young researcher grant. The work reflects only the authors' views. The Community in not liable for any use that may be made of the information contained therein. The authors thank Mrs. Vesna Mislej from Central Wastewater Treatment Plant in Ljubljana and Mrs. Marjetka Levstek from Wastewater treatment plant Domžale-Kamnik for providing wastewater samples.Peer reviewe