Agricultural intensification, and in particular the use of pesticides, leads over the years to a loss of biodiversity and a decline of ecosystem services in cultivated zones and agricultural landscapes. Among the animal communities involved in the functioning of agro-ecosystems, earthworms are ubiquitous and recognized as indicators of land uses and cultural practices. However, little data is available on the levels of pesticides in such organisms in natura, which would allow estimating their actual exposure and the potentially resulting impacts. Thus, the objective of this study was to develop a sensitive analytical methodology to detect and quantify 27 currently used pesticides in earthworms (Allolobophora chlorotica). A modified QuEChERS extraction was implemented on individual earthworms. This step was followed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The whole analytical method was validated on spiked earthworm blank samples, with regard to linearity (from 1 to 100 method limit of quantification, r2 > 0.95), intra-day precision (relative standard deviation (RSD) < 15%), inter-day precision (RSD < 20%), recoveries (mainly in the range 70–110%), and limits of detection and of quantification (inferior to 5 ng/g for most of the pesticides). The developed method was successfully applied to determine the concentrations of pesticides in nine individuals collected in natura. Up to five of the selected pesticides have been detected in one individual.
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The authors would like to thank the “Zone Atelier Plaine et Val de Sèvre” and especially Vincent Bretagnolle for the study site access and provision of site infrastructures. They thank J. Mathieu (University Pierre and Marie Curie, IEES, Paris) for the earthworm supply from its laboratory culture. They also thank all the people who took part in the earthworm sampling in Chizé. ZA PVS is a long-term research platform of the network Recotox (https://www.recotox.eu/). The authors are grateful to Recotox for supporting the study
This work has been financially supported by the French ONEMA within the national call “Pesticides” (APR ECOPHYTO 2014).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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