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To obtain the presence of environmental contaminants in honeybee and compare the toxicity of the detected pesticides to Apis mellifera ligustica Spin and Apis cerana cerana Fabricius. In this work, 214 honeybee samples were collected to simultaneous monitoring 66 pesticides between 2016 and 2017 in China. A modified QuEChERS extraction method coupled with multi-residue analytical methods by Ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and Gas chromatography-mass spectrum (GC–MS). Among, four pyrethroid pesticides were selected to test and compare the acute oral toxicities of two honeybees. And the survival risk of beta-cypermethrin was analyzed to them. Using this method, 21 compounds were detected, including 3 neonicotinoids, 5 pyrethroids, 5 organophosphorus and 8 others. Importantly, detected frequencies of pyrethroid pesticides were accounted for 53.3%. Among, acute toxicity values (LD50) of four pyrethroid pesticides to the A.m. ligustica were higher than of that the A.c. cerana. When they were exposed to the same concentration of beta-cypermethrin (0.2906 mg/L), the survival rate of the A.m. ligustica (40.0%) was higher than the A.c. cerana (18.9%). Our work is valuable to analyze multiple pesticide residues of honeybees and evaluate the survival risk of two honeybee species, which also provides a basis for the risk assessment.

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