Objective: To study the benzo (a) pyrene (BaP) induced DNA adduct in rat and establish a method to measure the DNA adduct in blood by high performance liquid chromatography (HPLC).
Methods: The SD rats were treated with 100 mg/kg of BaP-DMSO (cosolvent: 1% sodium carboxymethyl cellulose) once by i.p., and the blood of femoral vein was collected 5 hours later. The blood DNA was extracted by kit and confirmed by agarose gel electrophoresis. After extraction, the DNA adducts were hydrolyzed in 0.1 nmol/L HCl at 90 degrees C for 4 hours. The acid-hydrolysis products (BP-tetrols) of DNA adducts were extracted by ethyl acetate and measured by HPLC, and finally confirmed by HPLC-MS.
Results: In chromatogram there were new peaks to occur for rats treated by BaP, with compared to control. By HPLC-MS, one of the new peaks was confirmed to be BP-tetrol.
Conclusion: The rats ingesting BaP in the way described in this experiment can form DNA adducts, and the adducts in blood can be detected by HPLC.
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