There is growing concern over the environmental and human health impacts of chemical contaminants in agricultural systems. The environmental persistence of veterinary antibiotics applied to agricultural fields during manure fertilization could lead to increased antibiotic resistance. New generation, neonicotinoid pesticides pose a threat to aquatic ecosystem health due high water mobility and increased potential for non-target exposure. The objectives of this research were to develop a sensitive, analytical method for quantification of pirlimycin (PLY) in soils to be used in field research and determine the ability of second-generation neonicotinoids to move through soil when applied as a seed coating using a greenhouse study. Liquid-solid extraction of PLY from soil using (1:6, v/v) ammonium hydroxide/methylene chloride produced good PLY recovery (67-140%). Liquid-chromatography coupled with tandem mass-spectrometry for instrumental analysis provided good sensitivity with minimal matrix interferences. The mass balance distribution of neonicotinoid treatment coated onto corn seeds was determined in plant and soil samples for a single pot after 3 weeks of growth. A large percent (83-87%) of initial pesticide coating applied to seed was un-detected in plant in soil. Of the detected portion of neonicotinoid treatment, between 96-98% was observed to move out into the soil. This suggests the potential for long-range transport of seed-coated neonicotinoids.