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Biomarker responses and metabolites identification in Lumbricus terrestris exposed to drugs of environmental concern, an in vivo and in vitro approach

AuthorsSolé, Montserrat CSIC ORCID ; Montemurro, Nicola; Pérez Solsona, Sandra CSIC ORCID
Issue Date16-Oct-2020
PublisherConsejo Superior de Investigaciones Científicas (España)
Citation16th Annual Workshop On Emerging High-Resolution Mass Spectrometry (Hrms) And Lc-Ms/Ms Applications In Environmental Analysis And Food Safety : Abstract Book: 50 (2020)
AbstractThe earthworm Lumbricus terrestris, is a common anecic key species living in natural soils used also as bioindicator in soil pollution assessment. Previous research has demonstrated that earthworms are exposed to a variety of anthropogenic organic pollutants and they are able to metabolize organic soil contaminants such as polycyclic aromatic hydrocarbons, pesticides and pharmaceutical products. Specimens of L. terrestris were exposed in Petri dishes to filter paper embedded with 1 mg/mL to different model compounds for 48 h. The chemicals were: Lamotrigine, Cocaine, Fipronil (as RegentÔ800WG) and the organophosphorus pesticide bis-4-nitrophenyl phosphate (BNPP) used as positive control. At the end of this period, earthworms were immediately frozen in liquid N2 and their tissues, homogenized by cryogrinding, targeted to determine chemical exposure and metabolite formation as well as to biomarker determinations. The biomarkers analyzed were acetylcholinesterase (AChE), Glutathione-S-transferase (GST) and Carboxylesterase (CE) activities using different substrates. To study the accumulation and metabolism of tested compounds in earthworm tissues, analytes of interest were extracted using a modified QuEChERS apprach consisting of one single extraction step and analyzed using a SCIEX ExionLCTM AD system coupled to a hybrid SCIEX X500R QTOF system (Montemurro et al., 2020). High-resolution data were acquired in both positive and negative ESI combing a non-targeted screening workflow using SWATH acquisition which generates comprehensive and high-quality MS/MS spectra comparable to "MRM-like" fragments that can be used to confirm unequivocally the detection of specific compounds after comparing the SWATH data with pre-assembled MS/MS spectral libraries. The results obtained revealed the substrates 4-nitrophenyl butyrate (4-NPB) and 1-naphthyl butyrate (1-NB) in CE measures as the most responsive to cocaine and BNPP exposures. These results for BNPP corroborate the agreement between in vivo and in vitro responses. Chemical analysis and metabolite formation confirmed the extent of the exposure and identified the main metabolites formed for further in vivo exposures under more realistic conditions
Description16th Annual Workshop On Emerging High-Resolution Mass Spectrometry (Hrms) And Lc-Ms/Ms Applications In Environmental Analysis And Food Safety, 15-16 October 2020.-- 1 page
Appears in Collections:(IDAEA) Comunicaciones congresos
(ICM) Comunicaciones congresos
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