Analysis of DNA adducts bases by capillary electrophoresis with amperometric detection.

Abstract

We have developed a method for the detection of DNA adducts by combining capillary electrophoresis (CE) with the specificity of amperometric detection. Guanine is the most easily damaged base of the four normal DNA bases and many adducts of guanine have been found in DNA. These guanine adducts are often electrochemically active, while the normal bases with the exception of guanine are not. Therefore, CE with amperometric detection will be a promising method to study DNA damage. The four normal deoxynucleosides and two damaged deoxnucleosides N2-ethyldeoxyguanosine (N2-ethyl-dG) and 8-hydroxydeoxyguanosine (8-OH-dG), were completely separated by micellar electrokinetic chromatography (MEKC). Deoxyguanosine and the two damaged deoxynucleosides were identified using amperometric detection. The sensitivity of our system was comparable to that of UV detection. Analysis of DNA hydrolysis products was also performed briefly using this method.