title: HPLC method for quantification of oxidative stress by salicilate hydroxylation in human plasma.
abstract: The aim of the present study was to modify and validate a high-performance liquid chromatographic (HPLC) method for determining 2,3 and 2,5 dihydroxybenzoic 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 muL. 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. Inter- and intra-analysis variation coefficient was below 10%. Good recoveries of more than 99% were obtained for both metabolites using this method.
authors.affiliation: Universidad Autónoma de Zacatecas, Doctorado en Farmacología Médica y Molecular, Unidad Académica de Medicina Humana, Zacatecas, Zacatecas, México.
authors.forename: Blanca P
authors.initials: BP
authors.lastname: Lazalde-Ramos
authors.forename: Ismael
authors.initials: I
authors.lastname: Lares-Asseff
authors.forename: Ignacio
authors.initials: I
authors.lastname: Villanueva-Fierro
authors.forename: Martha
authors.initials: M
authors.lastname: Sosa-Macías
authors.forename: Patricia
authors.initials: P
authors.lastname: Yahuaca-Mendoza
authors.forename: Ana
authors.initials: A
authors.lastname: Zamora-Perez
mesh_terms: D015415:Biomarkers / Q000097:blood; D002396:Catechols / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005841:Gentisates / Q000097:blood*; D006801:Humans; D062385:Hydroxybenzoates / Q000032:analysis; D006900:Hydroxylation; D016014:Linear Models; D018384:Oxidative Stress; D015203:Reproducibility of Results; D012459:Salicylates / Q000737:chemistry / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: D015415:Biomarkers; D002396:Catechols; D005841:Gentisates; D062385:Hydroxybenzoates; D012459:Salicylates; C009091:protocatechuic acid; C009135:2,3-dihydroxybenzoic acid; C010925:2,5-dihydroxybenzoic acid