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

Csilla Mi�?�?�?�?�?�?�?�?�?�?�?�?�?�?�?¾anov�?�?�?�?�?�?�?¡, Anton Kebis

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.