Neonicotinoids (NNDs) constitute a major class of insecticides with a broad and versatile spectrum of applications in agriculture. Hence, their residues are found in several environmental compartments and can be transferred via several pathways to numerous organisms. Despite their profound impact on honeybees and wild bees (impairment of memory, impact on immune system), their presence in humans is far less reported, possibly due to the low to moderate toxicological effects that they elicit. The aim of the present review is to emphasize on developments in the biomonitoring of NNDs. It focuses mainly on chromatographic analysis of NNDs and their metabolites in human biological fluids, discussing key features, such as sample preparation and analytical method validation. Nonetheless, case reports regarding intoxication incidents are presented, highlighting the significance of such cases especially in the developing world.