Measurement of level of oxidative DNA damage in liver perfusate after pretreatment with copper solution using HPLC with electrochemical detection

Csilla MišÃÃ

Abstract

The aim of the present study was to develop a RP-HPLC method for the determination of a background level of 8-oxodG (a biomarker of oxidative stress) in rat liver after addition of copper to perfusate. The reversed phase analytical column PurospherSTARC18e (150x4.6mm, I.D., 5 m,Merck) with PurospherSTAR RP-18e (4x4mm, I.D., 5 m,Merck) as a precolumn were applied for the analysis. The mobile phase consisted of 8% (v/v) methanol in 50mmol/Lphosphate buffer, pH5.5.Oxidative damage to nuclear DNAwas determined by the simultaneousmeasuring of 2‘- deoxyguanosine (dG) with UV detection folowed by electrochemical detection of 8-oxodG. The validation of the HPLC method according to linearity, accuracy and precision was investigated. A detailed investigation of experiments have been discussed in this paper.

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