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The dataset represent the results of a cross-sectional study conducted in Telangana, India, to investigate the effects of pesticide exposure in agricultural regions. This study includes 341 pesticides exposed participants and 152 controls in three districts of Telangana including 15 farming villages. Blood and urine samples were analysed to determine various pesticide concentrations present in blood including organophosphates, carbamates and pyrethroids group of pesticides, and six dialkyl phosphate (DAP) metabolites, including dimethyl phosphate (DMP), diethyl phosphate (DEP), dimethyl thiophosphate (DMTP), dimethyldithiophosphate (DMDTP), diethylthiophosphate (DETP) and diethyldithiophosphate (DEDTP) were analysed in urine samples.In addition the enzyme acetylcholinesterase (AChE) activity were measured using advanced analytical methods. The data provide information on pesticide profiles, exposure biomarkers and the relationship between exposure duration and AChE activity. These study results emphasise the importance and addressing pesticide-related health problems in farmers for proactive measures to mitigate the harmful effects of pesticide exposure in agriculture., (© 2024 The Author(s).)