Micro QuEChERS-based method for the simultaneous biomonitoring in whole blood of 360 toxicologically relevant pollutants for wildlife.

This work presents the optimization, validation, and verification of a miniaturized method for the determination of 360 environmental pollutants that are of toxicological concern for wildlife. The method implies a one-step QuEChERS-based extraction of 250 μl whole blood using acidified acetonitrile, followed by two complementary analyses by LC-MS/MS and GC-MS/MS. The optimized conditions allow the simultaneous determination of the major persistent organic pollutants, a wide range of plant protection products, rodenticides, pharmaceuticals, and a suite of metabolites that can be used as biomarkers of exposure. The method is very sensitive, and 95% of the pollutants can be detected at concentrations below 1.5 ng/ml. The method was applied to a series of 148 samples of nocturnal and diurnal wild raptors collected during field ecological studies in 2018 and 2019. Fifty-one different contaminants were found in these samples, with a median value of 7 contaminants per sample. As expected, five of the six contaminants that were detected in >50% of the samples were persistent or semi-persistent organic pollutants. However, it is striking the high frequency of detection of some non-persistent pollutants, such as 2-phenylphenol, benalaxyl, metaflumizone, diphenylamine, brodifacoum or levamisole, indicating the penetration of these chemicals into the food chains. The toxicological significance of all these findings should be studied in depth in future research. However, the results clearly demonstrated that the approach developed provides reliable, simple, and rapid determination of a wide range of pollutants in wildlife and makes it very useful to obtain valuable data in biomonitoring studies with only small amounts of sample.