RECENT ADVANCES IN CHEMISTRY AND THE ENVIRONMENT Method for quantifying NSAIDs and clofibric acid in aqueous samples, lumpfish (Cyclopterus lumpus) roe, and zebrafish (Danio rerio) eleutheroembryos and evaluation of their bioconcentration in zebrafish eleutheroembryos N. Molina-Fernandez 1 & C. Perez-Conde 1 & S. Rainieri 2 & J. Sanz-Landaluze 1 Received: 15 January 2016 /Accepted: 10 April 2016 # Springer-Verlag Berlin Heidelberg 2016 Abstract Pharmaceuticals such as nonsteroidal anti- inflammatory drugs (NSAIDs) and lipid regulators are being repeatedly detected at low concentrations (pg · mL -1 –ng · mL -1 ) in the environment. A large fraction of these com- pounds are ionizable. Ionized compounds show different physico-chemical properties and environmental behavior in comparison to their neutral analogs; as a consequence, the quantification methods currently available, based on the neu- tral molecules, might not be suitable to detect the correspond- ing ionized compounds. To overcome this problem, we devel- oped a specific analytical method to quantify NSAIDs and lipid regulators (i.e., ibuprofen, diclofenac, naproxen, and clofibric acid) and their ionized compounds. This method is based on three steps: (1) the extraction of the organic com- pounds with an organic solvent assisted with an ultrasonic probe, (2) the cleaning of the extracts with a dispersive SPE with C 18 , and (3) the determination of the chemical com- pounds by GC-MS (prior derivatization of the analytes). We demonstrated that the proposed method can successfully quantify the pharmaceuticals and their ionized compounds in aqueous samples, lumpfish eggs, and zebrafish eleutheroembryos. Additionally, it allows the extraction and the cleanup of extracts from small samples (0.010 g of wet weight in pools of 20 larvae) and complex matrixes (due to high lipid content) and can be used as a basis for bioaccumu- lation assays performed with zebrafish eleutheroembryos in alternative to OECD test 305. Keywords Zebrafish eleutheroembryos . Bioconcentration . NSAIDs . Clofibric acid . Cleanup . GC-MS Introduction NSAIDs together with lipid regulators are among the most commonly applied analgesics and anti-inflammatory drugs worldwide (Macia et al. 2007). Within those, ibuprofen (IBU), diclofenac (DC), naproxen (NP), and clofibric acid (CA) are some of the most frequently used. As a result of their widespread use, these products and their metabolites and products of degradation can enter the water supply from do- mestic or industrial waste water discharges, commercial food treatment procedures, and the ground application of manure, and in recent years, they are being detected at low concentra- tions levels (range of pg- or ng · mL -1 ) in various environmen- tal media such as surface water and sediments (Azzouz et al. 2010; Azzouz et al. 2015; Gentili et al. 2012; Arroyo et al. 2011; Dubreil-Chéneau et al. 2011; Grueiro Noche et al. 2011; Zhang H et al. 2013; Sadat Hasheiminasab et al. 2013; Coimbra et al. 2015; Corcoran et al. 2015). There is evidence that NSAIDs and CA can induce ecotox- icological effects and bioaccumulation in aquatic organism (Corcoran et al. 2010). Coimbra et al. found a significant reduction in growth, a lower muscle triglyceride, impacts in male gonad development, and a decrease in the fecundity in adult zebrafish exposed at 10 mg · g -1 as well as abnormalities Responsible editor: Ester Heath Electronic supplementary material The online version of this article (doi:10.1007/s11356-016-6671-8) contains supplementary material, which is available to authorized users. * J. Sanz-Landaluze jsanzlan@ucm.es 1 Department of Analytical Chemistry, Faculty of Chemical Science, Complutense University of Madrid, Avenida Complutense s/n, 28040 Madrid, Spain 2 Food Research Division, AZTI, Parque Tecnológico de Bizkaia, Astondo Bidea 609, 48160 Derio, Spain Environ Sci Pollut Res DOI 10.1007/s11356-016-6671-8