The study on in vitro formation of DNA adduct 8-hydroxy-2-deoxyduanosine (8-OHdG) from benzo[a]pyrene and Fenton-like reaction

• Main Authors: Budiawan, B., Putri, D. O., Handayani, S., Dani, I. C., Bakri, R.
• Format: Conference Proceeding
• Language: English
• Subjects: Adducts; Biomarkers; Chromium; Deoxyribonucleic acid; DNA; Heavy metals; High performance liquid chromatography; Hydrogen peroxide; Reagents; Synergistic effect; Vitamin C
• ISSN: 0094-243X; 1551-7616
• DOI: 10.1063/1.5064107

Cigarette is one of the factors that increases the numbers of human deaths because it contains many harmful substances, such as benzo[a]pyren (B[a]P) and heavy metal Cr(VI). One of the consequences caused by both substances is an oxidative DNA damage and DNA adducts 8-hydroxy-2’-deoxyguanosine (8-OHdG) production as a biomarker of cancer risk. In this research, in vitro assay of benzo[a]pyrene with Fenton-like reagent was conducted in several pHs (7.4 and 8.4), temperatures (37 °C and 60 °C), and incubation times (5 and 7 hours). The 8-OHdG DNA adducts was analyzed using a reverse phase HPLC chromatography with UV/vis detector at the wavelength of 254 nm. The result shows that the 8-OHdG concentration due to exposure of B[a]P in the presence of Fenton-like reagent has exceeded the minimum concentration of detection that was set as LOD value of 5.188 ppb. Based on the data result, the increasing time and temperature incubation have a synergistic effect of the 8-OHdG-concentration increase. The highest concentration of 8-OHdG formation was conducted in the condition of pH 7.4, temperature 60°C, and incubation time 12 h in the presence of B[a]P, Cr(VI), H2O2 and vitamin C as much as 15.13 ppb.