A high performance liquid chromatography-electrochemical array method for the measurement of oxidative/nitrative changes in human urine.
Oxidative stress has been implicated in various pathologies as well as in environmental pollutant-induced negative health outcomes. In this work we have developed an analytical method to measure oxidative stress markers namely m-, o-tyrosine, 3-chlorotyrosine, 3-nitrotyrosine, and the DNA damage marker 8-hydroxy deoxyguanosine in human urine. The method involves the base hydrolysis of the urine sample followed by solid phase extraction of target analytes using a reverse phase polymeric sorbent column prior to the HPLC-EC analysis. The recovery studies indicated that the overall method recovery for all analytes were >60%. The limit of quantification of all target analytes was <10nM with a linear range from 2 nM to 150 microM. The applicability of this method is demonstrated by analyzing the above markers in healthy human urine (n=10) samples.
Mechanistic Studies Division, Environmental Health Science and Research Bureau, Health Canada, Ottawa K1A0K9, Canada.
Gurusankar
G
Saravanabhavan
Erica
E
Blais
Renaud
R
Vincent
Prem
P
Kumarathasan
D000080242:8-Hydroxy-2'-Deoxyguanosine; D015415:Biomarkers / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000652:urine; D055664:Electrochemical Techniques / Q000379:methods*; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016014:Linear Models; D008297:Male; D018384:Oxidative Stress / Q000502:physiology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000652:urine
D015415:Biomarkers; C002744:3-nitrotyrosine; D014443:Tyrosine; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine; C087259:3-chlorotyrosine