Measurement of oxidized nucleobases and nucleosides in human urine by using a GC/MS assay in the selective ion monitoring mode View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

1995

AUTHORS

M.-F. Incardona , F. Bianchini , A. E. Favier , J. Cadet

ABSTRACT

Urinary excretion of oxidized bases and nucleosides has been suggested as a non-invasive biomarker of oxidative damage and repair. Oxidative adducts, resulting from a variety of reactive oxygen species. are in fact excreted in urine following repair. A method for the analysis of 5-hydroxyuracil (5-OHUra), 5-hydroxy-2’-deoxyuridine (5-OHdUrd). 5-hy- droxymethyluracil (5-HMUra). 5-hydroxymethyl-2’-deoayuridine (5-HMdUrd) in urine has been developed, based on GC/MS quantification. Stable isotopically labelled (M + 4) analogues, synthesized in our laboratory, are used as internal standards, assuring appropriate quantification in complex biological fluids. Urines are first purified by HPLC on a reverse phase column and fractions corresponding to the compounds of interest derivatized for GC/MS analysis. Two different derivatives for GC/MS determination, the trimethylsylyl (TMS) and the tert-butyldimethylsylyl (t-RDMS) derivatives. have been tested. Detection limit in the selective ion monitoring (SIM) mode is generally two-fold lomer with t-BDMS. Preliminary results on four healthy subjects show that the oxidized bases 5-OHUra and 5-HMUra are excreted in urine (approximately 0.7 nmol/kg/day), while the corresponding nucleosides are not detectable. The developed method could have important applications in the analysis of oxidative lesions in DNA exposed to oxidative stress or in DNA from biological samples. More... »

PAGES

249-260

References to SciGraph publications

Book

TITLE

Analysis of Free Radicals in Biological Systems

ISBN

978-3-0348-9895-9
978-3-0348-9074-8

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-3-0348-9074-8_18

DOI

http://dx.doi.org/10.1007/978-3-0348-9074-8_18

DIMENSIONS

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