Bisphenol A (BPA) is an industrial chemical used in food contact materials such as canned coatings. Meanwhile Ni (II) is one of the metal compounds that can be found naturally in food and water, and might be increased due to industrial waste. Both of BPA and Ni (II) are categorized as carcinogenic compounds that have possibility to trigger cancer in human. This study was conducted to analyze the effects of BPA and Ni (II) exposure on the formation of DNA adduct 8-hydroxy-2'-deoxyguanosine (8-OHdG) as biomarker of risk cancer through in vitro reaction using 2'-deoxyguanosine. Our previous in vitro studies were conducted in physiological condition, while in this study we traced the formation of 8-OHdG by reacting 2'-deoxyguanosine with various mixture of BPA and Ni (II) in the presence of H2O2 as oxidizing agent at pH 8.4, temperature 60 °C, and two incubation time 7 and 12 hours. Analysis of 8-OHdG formation was carried out by using a UHPLC reverse phase with UV-Vis detector at 254 nm and C18 as a stationary phase. The results showed that the highest concentration of 8-OHdG was on samples that contains BPA and H2O2 with 12 hours incubation time, as much as 85.313 ppb.