Fluorescence polarization immunoassay for the determination of diclofenac in wastewater
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RESEARCH PAPER
Fluorescence polarization immunoassay for the determination of diclofenac in wastewater Anna Raysyan 1,2 & Robin Moerer 2 & Bianca Coesfeld 1 & Sergei A. Eremin 3 & Rudolf J. Schneider 1,4 Received: 5 February 2020 / Revised: 8 November 2020 / Accepted: 10 November 2020 # The Author(s) 2020
Abstract Pharmacologically active compounds are often detected in wastewater and surface waters. The nonsteroidal anti-inflammatory drug diclofenac (DCF) was included in the European watch list of substances that requires its environmental monitoring in the member states. DCF may harmfully influence the ecosystem already at concentrations ≤ 1 μg L−1. The fast and easy quantification of DCF is becoming a subject of global importance. Fluorescence polarization immunoassay (FPIA) is a homogeneous mix-and-read method which does not require the immobilization of reagents. FPIA can be performed in one phase within 20– 30 min, making it possible to analyse wastewater without any complicated pre-treatment. In this study, new tracer molecules with different structures, linking fluorophores to derivatives of the analyte, were synthesized, three homologous tracers based on DCF, two including a C6 spacer, and one heterologous tracer derived from 5-hydroxy-DCF. The tracer molecules were thoroughly assessed for performance. Regarding sensitivity of the FPIA, the lowest limit of detection reached was 2.0 μg L−1 with a working range up to 870 μg L−1. The method was validated for real wastewater samples against LC-MS/MS as reference method with good agreement of both methods. Keywords Diclofenac . FPIA . Water . Wastewater . NSAID
Introduction Diclofenac (DCF) is a nonsteroidal anti-inflammatory drug (NSAID) with analgesic, anti-inflammatory, and antipyretic properties. The mechanism of action of diclofenac, like that of other NSAIDs, involves inhibition of cyclooxygenase (COX-1 and COX-2). Another pharmacological effect is the inhibition of prostaglandin synthesis in vitro [1]. The disposal of DCF via treated wastewater may harmfully influence the ecosystem already at concentrations ≤ 1 μg L−1 [2]. Therefore, DCF was included in the first EU Water Framework Directive * Rudolf J. Schneider [email protected] 1
Bundesanstalt für Materialforschung und -prüfung (BAM), 12200 Berlin, Germany
2
Department of Chemistry, Humboldt-Universität zu Berlin, 10099 Berlin, Germany
3
Chemical Faculty, M.V. Lomonosov Moscow State University, Moscow, Russian Federation 119991
4
Technische Universität Berlin, Straße des 17. Juni 135, 10623 Berlin, Germany
Watch List for emerging water pollutants to be monitored in surface waters of the EU member states [2, 3]. Various instrumental methods have been applied for detection of DCF, including ultra-performance liquid chromatography (UPLC) coupled to high-resolution mass spectrometers [4, 5]. However, these techniques are expensive and require extensive sample preparation [6]. Immunoassays have been studied and used for the detection of DCF, and many research reports deal with polyclonal a
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