This in vitro study was conducted to analyze the formation of biomarker for DNA damage 8-hydroxy-2'-deoxyguanosine (8-OHdG) due to Bisphenol A and metal ion Cu (I) exposure in the presence of H2O2 as oxidizing agent on 2-deoxiguanosine via Fenton-Like reaction in non-physiological conditions (temperature 60°C and pH 8.4). Samples were incubated in numerous incubation time (7 hours and 12 hours) and further analyzed by using UHPLC reverse phase with mobile phase sodium phosphate buffer and methanol (85:15) with UV-Vis detector at wavelength 254 nm. The results showed that 8-OHdG can be formed by reacting Bisphenol A, Cu(I) and H2O2. Mostly, samples with longer incubation time (12 hours) have higher 8-OHdG concentration than shorter incubation time (7 hours). 8-OHdG concentrations in all samples were above LOD value and mostly were above LOQ value. Only one mixture that have 8-OHdG levels below LOQ value, i.e. dG, Cu(I), and H2O2. The highest 8-OHdG concentrations found on the complete mixture of reagents (dG, Bisphenol A, Cu(I), and H2O2) with longer incubation time (12 hours) with concentration equal to 92.438 ppb.