Development of a liquid chromatography/tandem mass spectrometry method to investigate the presence of biomarkers of DNA damage in urine related to red meat consumption and risk of colorectal cancer
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Date 2013-11-15
Status Peer reviewed
URI http://data.open.ac.uk/oro/document/195298
Volume 27
Issue 21
Abstract <b>RATIONALE</b> <br></br><br></br> The consumption of red meat is known to enhance the endogenous formation of N-nitroso compounds (NOCs), which are potent carcinogens. DNA damage related to NOCs, and hence, red meat has been detected in colorectal cells and in blood. We proposed to extend previous studies to a non-invasive approach for the detection of O<sup>6</sup>-carboxymethyl-guanine (O<sup>6</sup>CMG) and O<sup>6</sup>-carboxymethyl-2’-deoxiguanosine (O<sup>6</sup>CMdG) in urine in relation to red meat intake using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The presence of the adduct in urine samples either as free base or as 2’-deoxynucleoside could help in determining the repair mechanism involved when such lesions are produced. A non-invasive assessment of DNA adducts could also allow for large scale analyses in the population and cancer prevention dietary strategies. <br></br><br></br> <b>METHODS</b> <br></br><br></br> An LC-MS/MS method for quantitation of O<sup>6</sup>CMG and the corresponding nucleoside O<sup>6</sup>CMdG was developed. 24 hours urine samples, collected from healthy volunteers (N=12) on red meat and vegetarian diets, were analysed either by direct injection or after purification by solid phase extraction (SPE). A separate LC-MS/MS method for the quantitation of O<sup>6</sup>-methyl guanine (O<sup>6</sup>MeG) and O<sup>6</sup>-methyl-2’-deoxyguanosine (O<sup>6</sup>MedG), which are possible hydrolysis products forming during the samples pre-treatment, was also developed. <br></br><br></br> <b>RESULTS</b> <br></br><br></br> The developed LC-MS/MS method allowed the simultaneous measurement of O<sup>6</sup>CMG and O<sup>6</sup>CMdG. The limit of detection was 0.38 ng/mL for O<sup>6</sup>CMG and 0.18 ng/mL for O<sup>6</sup>CMdG. The direct injection analysis of the clinical samples showed low sensitivity due to high background signal that was improved by SPE purification. However, the concentrations of the adducts were still found to be below the LOD. <br></br><br></br> <b>CONCLUSIONS</b> <br></br><br></br> Two novel, reproducible, and accurate LC-MS/MS methods were developed for the determination of O<sup>6</sup>CMG and O<sup>6</sup>CMdG, and O<sup>6</sup>MeG and O<sup>6</sup>MedG in urine. Clinical samples from volunteers on different diets were analysed. Further studies are required to discover a link between the presence of these biomarkers in urine and red meat consumption.
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Label Da Pieve, Chiara ; Sahgal, Natasha ; Moore, Sharon A. and Velasco-Garcia, Maria N. (2013). Development of a liquid chromatography/tandem mass spectrometry method to investigate the presence of biomarkers of DNA damage in urine related to red meat consumption and risk of colorectal cancer. Rapid Communications in Mass Spectrometry, 27(21) pp. 2493–2503.
Title Development of a liquid chromatography/tandem mass spectrometry method to investigate the presence of biomarkers of DNA damage in urine related to red meat consumption and risk of colorectal cancer