Quantification of emerging micropollutants in an amphipod crustacean by nanoliquid chromatography coupled to masss pectrometry using multiple reaction monitoring cubed mode

An innovative analytical method has been developed to quantify the bioaccumulation in an amphipodcrustacean (Gammarus fossarum) of three micropollutants regarded as anthropic-pollution markers: car-bamazepine, oxazepam, and testosterone. A liquid-liquid extraction assisted by salts, known as QuEChERS(Quick, Easy, Cheap, Effective, Rugged, and Safe) was miniaturised and optimised, so it could be adaptedto the low mass samples (approximatively 5 mg dry weight). For this same reason and in order to obtaingood sensitivity, ultra-trace analyses were carried out by means of nanoliquid chromatography. A pre-concentration system by on-column trapping was optimised to increase the injection volume. In order toimprove both sensitivity and selectivity, the multiple reaction monitoring cubed mode analyses (MRM3)were carried out, validated and compared to the classic MRM. To the best of our knowledge, this is thefirst time that MRM3is coupled to nanoliquid chromatography for the analysis and detection of organicmicropollutants <300 Da. The optimised extraction method exhibited recoveries superior to 80%. Thelimits of quantification of the target compounds were 0.3, 0.7 and 4.7 ng/g (wet weight) for oxazepam,carbamazepine and testosterone, respectively and the limits of detection were 0.1, 0.3 and 2.2 ng/g (wetweight), respectively. The intra- and inter-day precisions were inferior to 7.7% and 10.9%, respectively,for the three levels of concentration tested. The analytical strategy developed allowed to obtain lim-its of quantification lower than 1 ng/g (wet weight) and to establish the kinetic bioconcentration ofcontaminants within G. fossarum