HPLC method for quantification of oxidative stress by salicilate hydroxylation in human plasma

Blanca P. Lazalde-Ramos, Ismael Lares-Asseff, Ignacio Villanueva-Fierro, Martha Sosa-Macías, Patricia Yahuaca-Mendoza, Ana Zamora-Perez

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4 Scopus citations

Abstract

The aim of the present study was to modify and validate a highperformance liquid chromatographic (HPLC) method for determining 2,3 and 2,5 di hydroxy benzoic acid (2,3-DHBA and 2,5-DHBA) from salicylic acid in human plasma. The mobile phase was a mixture of sodium acetate/citrate (pH 2.5) 30 mM-methanol (93:7, v/v). The injection volume was 10 μL. Retention time for 2,5-DHBA, and 2,3-DHBA was 4.5 ± 0.10 and 5.8 ± 0.15 min, respectively. The detection and quantification limits were 10 and 40 nM for 2,3-DHBA and 8 and 20 nM for 2,5-DHBA. Linearity was evaluated in the range of 40-1600 nM for both metabolites. Interand intra-analysis variation coefficient was below 10%. Good recoveries of more than 99% were obtained for both metabolites using this method.

Original languageEnglish
Pages (from-to)675-679
Number of pages5
JournalJournal of chromatographic science
Volume48
Issue number8
DOIs
StatePublished - Sep 2010

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