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Neonicotinoid insecticide use has been suggested as a cause of honey bee colony decline; however, detection rates and concentrations of neonicotinoid insecticide residues in field-collected honey bees have been low. We collected honey bee and beebread samples from apiaries in agricultural, developed, and undeveloped areas during 2 years in Virginia to assess whether landscape type or county pesticide use was predictive of honey bee colony exposure to neonicotinoid insecticides. Trace concentrations of the neonicotinoid imidacloprid were detected in honey bees (3 of 84 samples, 2.02-3.97 ng/g), whereas higher concentrations were detected in beebread (5 of 84 samples, 4.68-11.5 ng/g) and pollen (three of five pollen trap samples, 7.86-12.6 ng/g). Imidacloprid was only detected in samples collected during July and August and was not detected in honey bees from hives where neonicotinoids were detected in pollen or beebread. The number of hives sampled at a site, county pesticide use, and landscape characteristics were not predictive of neonicotinoid detections in honey bees or beebread (all P > 0.05). Field surveys may underestimate honey bee exposure to field-realistic levels of pesticides or the risk of exposure in different landscapes because of low detection rates. Undetectably low levels of exposure or high levels of exposure that go undetected raise questions with regard to potential threats to honey bees and other pollinators.

Three sample cleanup methods (silica SPE, NH2‐silica SPE, and Z‐Sep SPE) based on solid‐phase extraction were investigated for determination of neonicotinoid insecticides and selected metabolites in honey bee and pollen samples by liquid chromatography with tandem mass spectrometry. Samples were extracted by acidified hexane and ethyl acetate and then cleaned up with a solid‐phase extraction cartridge packed with silica gel, which showed a better cleanup efficiency compared to the aminopropyl silica solid‐phase extraction and zirconium‐based sorbents method. Matrix effects of the three cleanup methods were evaluated and compared. Silica gel showed the highest analyte recoveries and method detection limit for this method were 2.0 to 9.1 μg/kg for honey bees and 2.4 to 4.7 μg/kg for pollen. Recovery studies were performed at three spiking levels (10, 60, and 120 μg/kg) and ranged from 78 to 140% with relative standard deviations between 3 to 18% in honey bees and 83 to 124% with relative standard deviations between 3 to 17% in pollen. The silica gel solid‐phase extraction cleanup method was then applied using honey bee and pollen samples that were collected from different apiaries.