Analisis Adduct DNA Setelah Pemberian Natrium Nitrit dan Dimetilamin Secara Berulang Pada Tikus

Nitrosodimethylamine is a carcinogenic compound which can be formed from the reaction of nitrite and dimethylamine that is found in food. Nitrosodimethylamineis activated in liver and alkylates the DNA base and producing a DNA adductssuch as O6-methylguanine and N7-methylguanine that have a role incarcinogenesis. In this research, DNA was isolated from rat’s blood which waspreviously given nitrosodimethylamine’s precursor, sodium nitrite anddimethylamine. DNA adducts can be obtained from hydrolysis in hydrochloricacid 0.1 N for 30 minutes at 7000C. Then the adducts were analyzed using High Performance Liquid Chromatography (HPLC), with a strong cation exchangecolumn (Supelcosil LC-SCX, 5 μm, 250 x 4.6 mm), mobile phase consisting ofammonium phosphate with a final concentration of 40 mM, pH 3.00, flow rate 1.5mL/minute, column temperature 30oC and detected at exitation wavelength 286 nm and emission wavelength 366 nm. This method gave an acceptable validation result according to accuracy and precicion test results that fulfill the requirementand linear calibration curve with a quantitation limit of 22,5403 ng/mL. Rats were divided into six groups that two groups were given nitrosodimethylamine aspositive control, three groups were given prekursor, and the other was normalcontrol. Blood samples were collected in 1,2 and 4 hour after last induced. Aftergiving sodium nitrite 110 mg/kg bw and dimethylamine (1:5) orally for a week,N7-methylguanine and O6-methylguanine had not been detected in rat’s blood.