Determination of urinary 8-hydroxy-2′-deoxyguanosine in obese patients by HPLC with electrochemical detection

A simple amperometric detector with a platinum tubular electrode was tested for the determination of urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) by high-performance liquid chromatography. Analyte was extracted from human urine using a single-step solid-phase extraction with the average recovery of 71 +/- 7%. Analysis was carried out on a C-18 analytical column under isocratic conditions. The limit of detection of 8-OHdG of 3 nM (S/N = 3) was comparable with that achieved using the conventional amperometric detector. Procedure was used for the determination of 8-OHdG levels in 10 obese patients (body-mass index > 30 kg m(-2)) yielding the mean concentration 32 +/- 27.1 nM of 8-OHdG or 1.85 +/- 0.72 mumol 8-OHdG mol(-1) creatinine (4.63 +/- 1.8 mug 8-OHdG g(-1) creatinine), which is not much different from the levels found in healthy individuals. (C) 2004 Elsevier B.V. All rights reserved.