Artículos de revistas
HPLC Method for Quantification of Oxidative Stress by Salicilate Hydroxylation in Human Plasma
Fecha
2010-09Autor
Lazalde Ramos, Blanca Patricia
Lares Asseff, Ismael
Villanueva Fierro, Ignacio
Sosa Macías, Martha
Yahuaca Mendoza, Patricia
Zamora Perez, Ana
Institución
Resumen
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 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. Inter-and intra-analysis variation coefficient was below 10%. Good recoveries of more than 99% were obtained for both metabolites using this method.