Neonicotinoids (NEOs) are a class of broad-spectrum insecticides dominant in the global market. They were distributed extensively in the environment and occurred frequently in humans. Potential health effects of NEOs, such as neurological toxicity and diabetes to non-targeted mammals, have raised concerns. This review summarizes analytical methods of NEOs in human samples, their internal exposure levels and composition profiles in urine, blood, hair, breast milk, saliva and tooth samples with global comparisons, and daily NEOs exposure dose and relative health risks.Urinary NEOs levels in Asian populations were substantially higher than those in the U.S. and Europe, which may be due to different dietary patterns and insecticide applications across regions. N-desmethyl acetamiprid, 5-hydroxy-imidacloprid and olefin-imidacloprid were dominant among detected NEOs. NEO metabolites exhibited higher detection frequencies and levels than their parent compounds in humans, while investigations on NEO metabolites remain much limited. Current exposure assessments mainly focused on short-term urine analysis, while biomaterials for long-term monitoring, such as hair, nail and other alternatives, should also be considered. Large-scale epidemiological studies are critically needed to elucidate potential health outcomes associated with NEOs exposure.