In order to develop a tiered approach to identify relevant biomarkers and matrices for assessing pesticide exposure in residents living close to vineyards, five priority pesticides (boscalid, captan, folpel, mancozeb and tebuconazole) and their metabolites were analyzed in urine and hair samples from the biobank of a French national prevalence study conducted between 2014 and 2016. To do this, several analytical methods based on gas chromatography coupled with tandem mass spectrometry (GC/MS/MS) were developed by relying on the expertise of the laboratory and the scientific literature, in particular on a paper describing the use of gas chromatography-mass spectrometry for the determination in human urine samples of ethylene thiourea (ETU), a metabolite of mancozeb, after a supported liquid extraction followed by a derivatization step [1]. The main adaptations carried out as part of this study concerned:?the determination of ethylene urea (EU), another metabolite of mancozeb, at the same time as ETU in urine samples?the determination of all substances of interest including boscalid, EU and ETU, folpel and one of its metabolite (phthalimide), tebuconazole and one of its metabolite (hydroxytebuconazole), and tetrahydrophthalimide (metabolite of captan) in organic hair extracts by GC/MS/MS after a derivatization step.