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DOI: 10.1055/s-0035-1565786
Levels of markers for smoking and oxidative stress in the urine of Korean adults
Reactive oxygen species (ROS) can attack lipid, protein and nucleic acid simultaneously in the living cells. In nuclear and mitochondrial DNA, 8-hydroxydeoxyguanosine (8-OHdG) is the most frequently detected [1]. Oxidative stress is free radical-mediated damage caused by excess levels of ROS. Increased oxidative stress is associated with increased DNA damage, obesity, carcinogenicity, coronary heart disease and various chronic diseases. Smoking increases oxidative DNA damage by ˜50%. In this study, we performed biomonitoring for cotinine and 8-OHdG in 1000 Korean adults' urine by developed and validated method.
We developed simultaneous determination method of cotinine as marker of smoking and 8-OHdG as oxidative stress marker. 8-OHdG and cotinine in human urine were determined by LC-MS/MS (API 4000) techniques, after pretreatment by solid-phase extraction.
Linearity of calibration curve of target compounds were sufficient for analysis of these compounds at ppb level, with the coefficient of determination (r2) higher than 0.999. The accuracies was 93 to 105% and precision was lower than 10% RSD. Very strong positive correlation was found in the result of cross validation results of cotinine and 8-OHdG, performed by two laboratories. Cotinine and 8-OHdG were detected from all of the urine samples. The mean ± SD of 8-OHdG and cotinine were 4.46 ± 7.7, 292.8 ± 638.5 ng/mL, respectively. We found statistically significant positive correlation between urinary 8-OHdG and cotinine concentration (p < 0.001). The urinary concentrations of 8-OHdG were varied from general characteristics and life styles of subjects.
References:
[1] Chiou CC, Chang Py, Chan EC, Wu TL, Tsao KC, Wu JT. Urinary 8-hydroxydeoxyguanosine and its analogs as DNA marker of oxidative stress: development of an ELISA and measurement in both bladder and prostate cancers. Clin Chim Acta 2003; 334: 87 – 94