engleski

Development of an optimized sample preparation procedure for the determination of trace levels of pharmaceuticals in fish

In the last few decades, pharmaceuticals (PhACs) have become one of the most prominent groups of emerging environmental contaminants. Since significant levels of PhACs are frequently reported in the aquatic environment [1, 2], exposure of aquatic organisms to these highly biologically active substances might lead to bioaccumulation and adverse effects on non-target species [3]. Unfortunately, the current knowledge on the occurrence and fate of PhACs in aquatic biota is still rather scarce, primarily due to a lack of adequate analytical methods. The main challenges in the trace analysis of PhACs include the large number and physicochemical diversity of PhACs and the complexity of biotic matrices, which often require time-demanding and tedious sample preparation protocols [4]. In this work, a new multiresidue method for the determination of 44 PhACs and their metabolites in freshwater fish was developed. The emphasis was on the development of a robust and reliable sample preparation protocol, applicable to different lipid-rich fish tissue matrices. Various extraction solvents and cleanup techniques were examined in model experiments to optimize the key analytical parameters: analytical recovery, matrix effect, and reproducibility. The final method protocol involved ultrasound-assisted extraction of cryomilled muscle tissue with basified methanol in two cycles, followed by a three-step extract cleanup procedure, including cryoprecipitation, filtration, and solid-phase extraction. The analysis of final extracts was performed using an optimized multiresidue LC/MS-MS method which included separation of the target analytes on a Synergy Polar HPLC column and highly specific MRM detection. The quantitative determination was based on isotopically-labeled surrogates, which were added to the sample before the extraction. The developed method was applied to assess PhACs levels in feral freshwater fish from the Sava River.

biota ; farmaceutici ; LC-MS/MS ; priprema uzorka

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