A serially coupled stationary phase method for the determination of urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine by liquid chromatography ion trap tandem mass spectrometry

Oxidative attack to DNA is of particular interest since DNA modifications can lead to heritable mutations. The most studied product of DNA oxidation is 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG). While 8-oxodG determination in blood and tissue cells is prone to artifacts, its measurement in...

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Bibliographic Details
Published in:Redox biology 2013-01, Vol.1 (1), p.492-497
Main Authors: Rota, Cristina, Cristoni, Simone, Trenti, Tommaso, Cariani, Elisabetta
Format: Article
Language:English
Subjects:
8-oxo-7,8-dihydro-2′-deoxyguanosine
Adult
Cations
Chromatography, Liquid - methods
Creatinine - urine
Deoxyguanosine - analogs & derivatives
Deoxyguanosine - analysis
Deoxyguanosine - urine
Female
Humans
Ion trap
LC-MS/MS
Male
Middle Aged
Oxidative stress
Reactive oxygen species
Reproducibility of Results
Research Paper
Tandem Mass Spectrometry - methods
Urine
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Summary:Oxidative attack to DNA is of particular interest since DNA modifications can lead to heritable mutations. The most studied product of DNA oxidation is 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG). While 8-oxodG determination in blood and tissue cells is prone to artifacts, its measurement in urine employing liquid chromatography tandem mass spectrometry (LC-MS/MS) has gained more and more interest for increased reliability. LC-MS/MS can be affected by matrix effects and this is particularly true when ion trap is used as MS analyzer, due to ion accumulation in the trap and related space charge effect. In the present work, we have developed a LC-MS/MS method where the combination of cation exchange and reverse phase solid phases resulted in LC separation optimization. This together with the employment of an isotopically labeled internal standard, allowed the usage of ion trap LC-MS/MS, typically not employed for quantitative measurement in biological samples, for the measurement of 8-oxodG in urine samples from control populations. Four different urine matrices were employed for method validation. Limit of quantitation was set at least at 0.5 ng/ml. While analyzing urine samples from healthy volunteers, 8-oxodG levels reported as ng/ml were statistically different comparing males with females (p
ISSN:2213-2317
2213-2317
DOI:10.1016/j.redox.2013.10.001