Simultaneous determination of five oxidative DNA lesions in human urine.
A method, involving a HPLC prepurification followed by a GC/MS analysis, has been set up for the measurement of nucleic acid oxidation products in human urine. For this purpose, isotopically labeled internal standards have been prepared and used for isotope dilution mass spectrometric detection. Using this approach, four oxidized DNA bases, i.e., 5-hydroxyuracil, 5-(hydroxymethyl)uracil, 8-oxo-7,8-dihydroadenine, and 8-oxo-7,8-dihydroguanine, together with 8-oxo-7,8-dihydro-2'-deoxyguanosine have been simultaneously quantified in human urine samples. The levels of the oxidized nucleic acid constituents, as expressed in picomoles per milliliter, were determined to be, in decreasing order: 8-oxo-7,8-dihydroguanine (583 +/- 376) > 5-(hydroxymethyl)uracil (121 +/- 56) > 5-hydroxyuracil (58 +/- 23) > 8-oxo-7,8-dihydro-2'-deoxyguanosine (30 +/- 15) > 8-oxo-7,8-dihydroadenine (7 +/- 4). Attempts to determine the amount of 5,6-dihydroxy-5,6-dihydrothymine, 5-hydroxycytosine, and 2,6-diamino-4-hydroxy-5-formamidopyrimidine using the above HPLC-GC/MS method were unsuccessful.
Ravanat
J L
JL
Laboratoire "Lésions des Acides Nucléiques", Service de Chimie Inorganique et Biologique, Département de Recherche Fondamentale sur la Matière Condensée, CEA Grenoble, 17 Avenue des Martyrs, F-38054 Grenoble Cedex 9, France.
Guicherd
P
P
Tuce
Z
Z
Cadet
J
J
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D003839:Deoxyadenosines / Q000652:urine; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000652:urine; D008401:Gas Chromatography-Mass Spectrometry; D006147:Guanine / Q000031:analogs & derivatives / Q000652:urine; D006801:Humans; D007553:Isotope Labeling; D009705:Nucleosides / Q000737:chemistry / Q000652:urine*; D010084:Oxidation-Reduction; D010432:Pentoxyl / Q000031:analogs & derivatives / Q000652:urine; D015203:Reproducibility of Results
D003839:Deoxyadenosines; D009705:Nucleosides; C008055:5-hydroxymethyluracil; C024829:8-hydroxyguanine; D006147:Guanine; C078251:2'-deoxy-7,8-dihydro-8-oxoadenosine; D010432:Pentoxyl; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine