A high performance liquid chromatography-electrochemical array method for the measurement of oxidative/nitrative changes in human urine.

Gurusankar Saravanabhavan, Erica Blais, Renaud Vincent, Prem Kumarathasan, Gurusankar Saravanabhavan, Erica Blais, Renaud Vincent, Prem Kumarathasan

Index: J. Chromatogr. A. 1217 , 3269-3274, (2010)

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Abstract

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.Crown Copyright (c) 2010. Published by Elsevier B.V. All rights reserved.