Enrico Capodicasa
Italy
Research Article
Free and Total Malondialdehyde Measured as 2,4-dinitrophenylhydrazine Adduct by HPLC-UV in Hemodialysis Patient Serum
Author(s): Roberto Biondi, Stefano Brancorsini, Maria Giulia Egidi, Giulia Poli, Enrico Capodicasa, Isabella Tritto, Gian Carlo Di Renzo, Florentina Duica, Dragos Cretoiu and Nicolae SuciuRoberto Biondi, Stefano Brancorsini, Maria Giulia Egidi, Giulia Poli, Enrico Capodicasa, Isabella Tritto, Gian Carlo Di Renzo, Florentina Duica, Dragos Cretoiu and Nicolae Suciu
Objective: Malondialdehyde (MDA) is an index of oxidative stress. In biological matrices MDA exists in both free (f-MDA) and bound (b-MDA) forms. In this report, a method to detect f-MDA and t-MDA was developed in human serum, based on MDA derivatization with 2,4-dinitrophenylhydrazine (DNPH) and HPLC separation. Methods: This method gave high sensitivity, with Limits of Detection (LOD) and Limits of Quantification (LOQ) of 3.5 pmol/ml and 10 pmol/ml, respectively; while recoveries of t-MDA from spiked matrices (R%) reached 98.1 ± 1.8 and 96.51 ± 1.8 for f-MDA. Considering the lower levels of f-MDA in biological fluids, this method improves derivatization conditions (DNPH medium solution, time and temperature) and yielded 47 ± 12 pmol/ml of f-MDA in serum of 17 controls. Our method succeeded to determine f-MDA and t-MDA in hemodialysed patients. Results: Results indicat.. View More»
