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Methodology for urinary 8-hydroxy-2'-deoxyguanosine analysis by HPLC with electrochemical detection
Bruno Spinosa De Martinis1, Maria de Lourdes Pires Bianchi
1Department of Pathology, Center of Legal Medicine, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Rua Tenente Catão Roxo 2418, Ribeirão Preto SP14051-140, Brazil. martinis@spider.usp.br
Abstract:
The compound 8-hydroxy-2'-deoxyguanosine (8-OHdG), one of the major oxidative DNA base products, is a commonly used biomarker of oxidative DNA damage. The routine measurement of 8-OHdG in biologic samples, and especially in urine, is inherently a challenging analytical problem due to the low levels of the analyte and the complexity of the sample. In this study, a simple and rapid procedure based on one step solid phase extraction (SPE) followed by high-performance liquid chromatography with electrochemical detection (HPLC-ECD) analysis is described. The methodology proposed resulted in a recovery of 74.5+/-12% for urinary 8-OHdG with a detection limit of 5.0 microg l(-1).