title: Detection and quantification of 8-hydroxy-2'-deoxyguanosine in Alzheimer's transgenic mouse urine using capillary electrophoresis.
abstract: 8-Hydroxy-2'-deoxyguanosine (8-OHdG) is one of the major forms of oxidative DNA damage, and is commonly analyzed as an excellent marker of DNA lesions. The purpose of this study was to develop a sensitive method to accurately and rapidly quantify the 8-OHdG by using CE-LIF detection. The method involved the use of specific antibody to detect the DNA lesion (8-OHdG) and consecutive fluorescence labeling. Next, urinary 8-OHdG fluorescently labeled along with other constituents were resolved by capillary electrophoretic system and the lesion of interest was detected using a fluorescence detector. The limit of detection was 0.18 fmol, which proved sufficient sensitivity for detection and quantification of 8-OHdG in untreated urine samples. The relative standard deviation was found to be 11.32% for migration time and 5.52% for peak area. To demonstrate the utility of this method, the urinary concentration of 8-OHdG in an Alzheimer's transgenic mouse model was determined. Collectively, our results indicate that this methodology offers great advantages, such as high separation efficiency, good selectivity, low limit of detection, simplicity and low cost of analysis.
authors.affiliation: Department of Neurosciences, UCSD School of Medicine, La Jolla, CA 92093, USA. chz002@ucsd.edu
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.forename: Gergana
authors.initials: G
authors.lastname: Nestorova
authors.forename: Robert A
authors.initials: RA
authors.lastname: Rissman
authors.forename: June
authors.initials: J
authors.lastname: Feng
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000544:Alzheimer Disease / Q000235:genetics / Q000652:urine*; D000818:Animals; D015415:Biomarkers / Q000652:urine; D001881:Borates; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000652:urine; D004195:Disease Models, Animal; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D018384:Oxidative Stress / Q000502:physiology; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; D001881:Borates; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine
keywords: 8-Hydroxy-2′-deoxyguanosine; Alzheimer's disease; Capillary electrophoresis laser-induced fluorescence detection; Oxidative DNA damage; Urine