title: Rapid measurement of 8-oxo-7,8-dihydro-2'-deoxyguanosine in human biological matrices using ultra-high-performance liquid chromatography-tandem mass spectrometry.
abstract: Interaction of reactive oxygen species with DNA results in a variety of modifications, including 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), which has been extensively studied as a biomarker of oxidative stress. Oxidative stress is implicated in a number of pathophysiological processes relevant to obstetrics and gynecology; however, there is a lack of understanding as to the precise role of oxidative stress in these processes. We aimed to develop a rapid, validated assay for the accurate quantification of 8-oxodG in human urine using solid-phase extraction and ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and then investigate the levels of 8-oxodG in several fluids of interest to obstetrics and gynecology. Using UHPLC-MS/MS, 8-oxodG eluted after 3.94 min with an RSD for 15 injections of 0.07%. The method was linear between 0.95 and 95 nmol/L with LOD and LOQ of 5 and 25 fmol on-column, respectively. Accuracy and precision were 98.7-101.0 and <10%, respectively, over three concentrations of 8-oxodG. Recovery from urine was 88% with intra- and interday variations of 4.0 and 10.2%, respectively. LOQ from urine was 0.9 pmol/ml. Rank order from the greatest to lowest 8-oxodG concentration was urine>seminal plasma>amniotic fluid>plasma>serum>peritoneal fluid, and it was not detected in saliva. Urine concentrations normalized to creatinine (n=15) ranged between 0.55 and 1.95 pmol/μmol creatinine. We describe, for the first time, 8-oxodG concentrations in human seminal plasma, peritoneal fluid, amniotic fluid, and breast milk, as well as in urine, plasma, and serum, using a rapid UHPLC-MS/MS method that will further facilitate biomonitoring of oxidative stress.
authors.affiliation: Oxidative Stress Group, Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK.
authors.forename: Patricia M W
authors.initials: PM
authors.lastname: Lam
authors.forename: Vilas
authors.initials: V
authors.lastname: Mistry
authors.forename: Timothy H
authors.initials: TH
authors.lastname: Marczylo
authors.forename: Justin C
authors.initials: JC
authors.lastname: Konje
authors.forename: Mark D
authors.initials: MD
authors.lastname: Evans
authors.forename: Marcus S
authors.initials: MS
authors.lastname: Cooke
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000328:Adult; D000653:Amniotic Fluid / Q000737:chemistry; D001202:Ascitic Fluid / Q000737:chemistry; D015415:Biomarkers / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000652:urine; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008895:Milk, Human / Q000737:chemistry; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D012463:Saliva / Q000737:chemistry; D012661:Semen / Q000737:chemistry; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D015415:Biomarkers; D017382:Reactive Oxygen Species; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine