Development of an HPLC method with electrochemical detection of femtomoles of 8-oxo-7,8-dihydroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine in the presence of uric acid

TALANTA

卷:63

Article

Rebelo, IA ; Piedade, JAP ; Oliveira-Brett, AM

关键词: DNA damage;    oxidative stress;    8-oxoGua;    8-oxodGuo;    8-oxo-7,8-dihydroguanine;    8-oxo-7,8-dihydro-2'-deoxyguanosine;    biomarker;    uric acid;    HPLC-ECD;    SPE;    urine;   

DOI  :  10.1016/j.talanta.2003.10.051

来源: Elsevier

【 摘 要 】

A selective method based on high performance liquid chromatography with electrochemical detection (HPLC-ECD) was developed to enable simultaneous detection of 8-oxo-7,8-dihydroguanine (8-oxoGua) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo), products of DNA oxidative damage, in the presence of uric acid (UA), a strong interferent in their electrochemical detection. The method developed consists of HPLC isocratic elution with amperometric detection oil a glassy carbon electrode, enabling a detection limit for 8-oxoGua and 8-oxodGuo lower than I nM in standard mixtures. Detection of low concentrations Lip to 25 nM of 8-oxoGua and 8-oxodGuo in the presence of UA in a 10(4)-fold higher concentration was achieved after one-step solid phase extraction (SPE). The method was tested with urine samples and it was possible to detect and quantify the presence of 8-oxoGua, and to confirm that UA was eliminated after uricase degradation and SPE. The LOD found in urine samples was about 80 nM, a Value higher than in standard mixtures, due to the increase of background current in the urine matrix. The results presented here contribute to the development of a methodological approach to simultaneous determination of 8-oxoGua and 8-oxodGuo ill urine samples. (C) 2003 Elsevier B.V. All rights reserved.

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