Abstract Pesticides have emerged as a global threat to human health, especially in developing countries. In this study, we examined nine major types of pesticide residues in the food commodities (8 groups) and serum samples (n = 80) of people living in the region of Southern-Punjab, Pakistan. Pesticide residues quantification was performed using gas chromatograph coupled with electron capture detector (GC-ECD). The estimated daily intakes of the selected pesticide residues through consumption of food were also calculated. The concentration of analyzed pesticide residues in food was found below the EU maximum permissible limits except for α-HCH, and β-HCH. The highest risk of cancer was found to be associated with the consumption of HCHs contaminated cereals than other products. The incidence rate of pesticide residues in the serum samples of studied subjects was 96.25%. Pearson correlation test showed no significant correlation between thyroid hormone levels of the studied population group and the analyzed pesticide residues in the food commodities (p>0.05). Findings of this study indicate that people of Southern-Punjab, Pakistan are exposed to pesticides via food chain although non-significant health implications were recorded, the levels of a few pesticides need to be curtailed.