HPLC method for quantification of oxidative stress by salicilate hydroxylation in human plasma.
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.
Lazalde-Ramos
Blanca P
BP
Universidad Autónoma de Zacatecas, Doctorado en Farmacología Médica y Molecular, Unidad Académica de Medicina Humana, Zacatecas, Zacatecas, México.
Lares-Asseff
Ismael
I
Villanueva-Fierro
Ignacio
I
Sosa-Macías
Martha
M
Yahuaca-Mendoza
Patricia
P
Zamora-Perez
Ana
A
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
D015415:Biomarkers; D002396:Catechols; D005841:Gentisates; D062385:Hydroxybenzoates; D012459:Salicylates; C009091:protocatechuic acid; C009135:2,3-dihydroxybenzoic acid; C010925:2,5-dihydroxybenzoic acid