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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

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.

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DOI (Digital Object Identifier) Link: https://doi.org/10.1002/rcm.6709
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Abstract

RATIONALE

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 O6-carboxymethyl-guanine (O6CMG) and O6-carboxymethyl-2’-deoxiguanosine (O6CMdG) 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.

METHODS

An LC-MS/MS method for quantitation of O6CMG and the corresponding nucleoside O6CMdG 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 O6-methyl guanine (O6MeG) and O6-methyl-2’-deoxyguanosine (O6MedG), which are possible hydrolysis products forming during the samples pre-treatment, was also developed.

RESULTS

The developed LC-MS/MS method allowed the simultaneous measurement of O6CMG and O6CMdG. The limit of detection was 0.38 ng/mL for O6CMG and 0.18 ng/mL for O6CMdG. 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.

CONCLUSIONS

Two novel, reproducible, and accurate LC-MS/MS methods were developed for the determination of O6CMG and O6CMdG, and O6MeG and O6MedG 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.

Item Type: Journal Item
Copyright Holders: 2013 John Wiley & Sons, Ltd.
ISSN: 0951-4198
Project Funding Details:
Funded Project NameProject IDFunding Body
Non-invasive biomarkers of DNA damage related to red meat intake and risk of colorectal cancerGrant 2007/52-R09World Cancer Research Fund
Keywords: DNA damage; red meat; colorectal cancer; biomarkers
Academic Unit/School: Faculty of Science, Technology, Engineering and Mathematics (STEM) > Life, Health and Chemical Sciences
Faculty of Science, Technology, Engineering and Mathematics (STEM)
Item ID: 38390
Depositing User: Maria Velasco
Date Deposited: 11 Oct 2013 09:11
Last Modified: 08 May 2019 13:31
URI: http://oro.open.ac.uk/id/eprint/38390
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