Pesticides are a major problem not only in the food chain but also when considering the protection of our planet. The use of neonicotinoids has been banned in the EU due to their high toxicity to living organisms, in particular honeybees. The presence of neonicotinoids in natural waters poses a threat to pollinating insects and thus hampers organic production. Pesticide residues in the natural waters of agricultural land are monitored within the framework of promoting sustainable rural development to maintain the safety of human and animal health. Chromatographic analyses of selected neonicotinoid pesticides in water samples from agricultural sites in eastern Poland were performed using high performance liquid chromatography (HPLC), which followed a solid-phase extraction (SPE). The research revealed no evidence of neonicotinoids contamination. Water quality in this region can be a good factor in promoting sustainable development. The obtained results complement the existing knowledge on the impact of neonicotinoids on both the sustainable food chain and the environment. Based on the results obtained, it is possible to conclude that they are not being used in rural area under the study.