Abstract
This study assesses the extraction of eleven pharmaceuticals, five pesticides, five perfluoroalkyl substances, and two illicit drugs in hemolymph from (Mytilus Galloprovincialis). Four extraction procedures using Phree™ Phospholipid Removal cartridges were tested using different volumes of methanol (400 and 600μL) and acetonitrile (300 and 450μL). The pollutants were determined by high-performance liquid chromatography-tandem mass spectrometry. The use of methanol gave several problems during the extraction procedure, such as longer times and sample loss. Three methods (acetonitrile 300 and 450μL; and methanol 600μL) were validated. Recoveries at three concentration levels (5, 50, and 100ng/mL) ranged 35.1-129.0 and 29.3-133.0% for acetonitrile 300 and 450μL, respectively, while recoveries for methanol 600μL ranged 52.2-166.0%. Limits of detection were<10ng/mL for most analytes using any of the methods. Methanol 600μL was the only method capable to extract the illicit drug 4-methoxyphencyclidine and provided a better peak shape and higher signal-noise ratio. When applied to non-spiked samples from local markets salicylic acid and diclofenac were detected at 33.50-97.79 and 28.30-30.31ng/mL respectively. To our knowledge, there are no methods to determine organic contaminants in hemolymph and this is the first application of Phree™ cartridges for mussel hemolymph extraction.
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