Abstract
Etheno-DNA adducts are generated from the metabolism of exogenous carcinogens and endogenous lipid peroxidation. We and others have previously reported that 1,N6-ethenodeoxyadenosine (εdA) and 3,N4-ethenodeoxycytidine (εdC) are present in human urine and can be utilized as biomarkers of oxidative stress. In this study, we report a new ultrasensitive UPLC-ESI-MS/MS method for the analysis of εdA and εdC in human urine, capable of detecting 0.5 fmol εdA and 0.3 fmol εdC in 1.0 mL of human urine, respectively. For validation of the method, 20 human urine samples were analyzed, and the results revealed that the mean levels of εdA and εdC (SD) fmol/µmol creatinine are 5.82 ± 2.11 (range 3.0–9.5) for εdA and 791.4 ± 328.8 (range 116.7–1264.9) for εdC in occupational benzene-exposed workers and 2.10 ± 1.32 (range 0.6–4.7) for εdA and 161.8 ± 200.9 (range 1.8–557.5) for εdC in non-benzene-exposed workers, respectively. The ultrasensitive detection method is thus suitable for applications in human biomonitoring and molecular epidemiology studies.
Highlights
Etheno-DNA adducts 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine are formed from exogenous carcinogens vinyl chloride and urethane [1,2,3,4], and peroxidation of arachidonic acid and liver microsomal membranes in the presence of lipid peroxidation (LPO)-inducing compounds [5–7]
Nair and others have established methods to study the formation and excretion of etheno-DNA base adducts; dA and dC were found to be present in human urine, and these could be biomarkers for DNA damage produced by persistent oxidative stress and lipid peroxidation [15,16,17]
For validation of the method, 20 human urine samples randomly selected from female workers who participated in a cross-sectional epidemiology study of occupational benzene exposure were assayed, and the results indicate that our ultrasensitive method appears to be superior to other methods reported in terms of: (1) high sensitivity and specificity; (2) low amounts of urine sample required; (3) capability to detect background levels of etheno-DNA adducts in human biological samples; and (4) reliable monitoring of the disease-related increase of these substances in patients
Summary
Etheno-DNA adducts 1,N6-ethenodeoxyadenosine ( dA) and 3,N4-ethenodeoxycytidine ( dC) are formed from exogenous carcinogens vinyl chloride and urethane [1,2,3,4], and peroxidation of arachidonic acid and liver microsomal membranes in the presence of LPO-inducing compounds [5–. Non-invasive detection methods, such as urinalysis, will expedite studies in humans aimed to elucidate etiopathological factors that cause oxidative DNA damage For these reasons, Nair and others have established methods to study the formation and excretion of etheno-DNA base adducts; dA and dC were found to be present in human urine, and these could be biomarkers for DNA damage produced by persistent oxidative stress and lipid peroxidation [15,16,17]. Participated in a cross-sectional epidemiology study of occupational benzene exposure were assayed, and the results indicate that our ultrasensitive method appears to be superior to other methods reported in terms of: (1) high sensitivity and specificity; (2) low amounts of urine sample required; (3) capability to detect background levels of etheno-DNA adducts in human biological samples; and (4) reliable monitoring of the disease-related increase of these substances in patients. We present the details of the method protocol and its application to quantify dA and dC in human urine samples
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