In-vitro STUDY OF ACRYLAMIDE AND TBHQ COMPOUNDS ON THE GENERATION OF DNA ADDUCT 8-HYDROXY-2’-DEOXYGUANOSINE AS BIOMARKER OF OXIDATIVE DNA DAMAGE

The research on DNA adduct 8-OHdG formation is performed by reacting 2'-deoxyguanosine and H2O2 with acrylamide (AA) and TBHQ compounds that were exposed to UVA light. The 8-OHdG forming test was conducted at various incubation times (5 h and 7 h) and pH variations (7.4 and 8.4). The 8-OHdG formation was analyzed using a reversed-phase HPLC instrument using a buffer solution of sodium phosphate pH 6.7: methanol (85:15) as eluent. The results showed that samples containing TBHQ and acrylamide (AA) produced 8-OHdG and shows synergistic and antagonistic effects. The 8-OHdG level of mixing acrylamide (AA) and TBHQ compounds increased significantly when exposed to UVA light. Samples incubated with UVA exposure had higher levels of 8-OHdG than samples without UVA exposure. The longer the incubation time, the greater the 8-OHdG content was produced. At pH 8.4, most of the samples resulted in greater 8-OHdG formation when compared to samples with the same mixture at pH 7.4. Samples with the greatest 8-OHdG formation results were found in samples containing 2'-deoxyguanosine, acrylamide (AA), TBHQ, and H2O2 at pH 8.4 with UVA light irradiation for 7h.