Despite increasing debates about their potential side effects on human health, data concerning the risks and the impacts associated with pesticides remains scarce. Analytical tools allowing the measurement of most pesticides and/or their metabolites to which the population can be exposed are also of need. In the present study, the limits of detection (LODs) of 3 different screening procedures based on either Low-Resolution and High-Resolution Mass Spectrometry (LR-MS and HR-MS) for the determination of pesticides in serum (among which carbamates, dithiocarbamates, neonicotinoids, organochlorines, organophosphates and pyrethroids) were explored. For HR-MS, a quadrupole time-of-flight was used in positive and negative electrospray ionization modes and data were obtained from either a targeted scheduled MSMS acquisition (HR-MSMS) or a data-independent acquisition (HR-DIA). For LR-MS, a triple quadrupole was used and data were acquired with a classical multiple-reaction monitoring (MRM) mode. Depending on the approach, the LOD values varied from 0.05 to 10 ng/mL. For the lowest concentrations, the proportion of molecules detected was systematically greater for the LR-MS approach, while those of HR-MSMS were better than those of HR-DIA. These differences in the LOD values were confirmed in a sample of 174 serums in which LR-MRM detected 89 compounds while HR-MSMS and HR-DIA detected 79 and 75 compounds, respectively. Nevertheless, for environmental and occupational purposes, HRMS approach could probably be efficient to detect most of pesticides and their metabolites in human serum and could be suitable for human biomonitoring studies or fundamental research exploring the impact of exposure to pesticides on human health.