Neonicotinoid insecticides (NEOs) have been extensively approved for application in agricultural production both in China and globally. Previous studies indicated that human exposure to NEOs would have adverse health effects. However, studies on occurrences of NEOs in kidney injury patients and their association with nephrotoxicity are limited. This study investigates the potential correlation between exposure to NEOs and kidney injury within a subset (n = 224) of the population residing in South China. Levels of 8 NEOs and 5 metabolites were quantified in blood samples obtained from both the 110 healthy and 114 kidney injury cohorts in South China. We found that target analytes were frequently detected in samples from the healthy and kidney injury cohorts (61%–100 %). Dinotefuran is the predominant NEO in the healthy (42.3 %) and kidney injury cohort (49.6 %). Significantly positive (p