Free and bound malondialdehyde measured as thiobarbituric acid adduct by HPLC in serum and plasma

• Published in: Clinical chemistry (Baltimore, Md.) Vol. 37; no. 8; pp. 1423 - 1429
• Main Authors: Carbonneau, MA, Peuchant, E, Sess, D, Canioni, P, Clerc, M
• Format: Journal Article
• Language: English
• Published: England Am Assoc Clin Chem 01.08.1991
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Female; Humans; Lipid Peroxidation; Male; Malondialdehyde - blood; Middle Aged; Neoplasms - blood; Renal Dialysis; Thiobarbiturates - blood; Thiobarbiturates - chemistry
• DOI: 10.1093/clinchem/37.8.1423
• ISSN: 0009-9147

Assay of free and total malondialdehyde (MDA) in human serum and plasma from healthy subjects and from patients with high risk of lipoperoxidation was performed as follows: (a) acidic (HClO4, pH 1, at 20 degrees C) or basic (NaOH, pH 13, at 60 degrees C) treatments for 30 min; (b) reaction of the protein-free extract (obtained by acid precipitation) with thiobarbituric acid (TBA); (c) HPLC separation on C18 columns with an eluting solution of methanol/phosphate buffer, 10 mmol/L, pH 5.8 (40/60, by vol), at a flow rate of 1.5 mL/min. Free MDA averaged 0.042 (SEM 0.008) and 0.043 (SEM 0.007) mumol/L, respectively, in serum and plasma from healthy subjects. Free (+/- SEM) MDA increased significantly in the plasma from cancer patients (0.270 +/- 0.047 mumol/L) and from hemodialyzed patients (0.214 +/- 0.035 mumol/L). In serum of hemodialyzed patients, analyses for total MDA were unsuitable because of interfering peaks. MDA bound to NH2 groups constituted 83.2% and 83.5% of total MDA in serum and plasma of healthy subjects, respectively, and only 58% in plasma of hemodialyzed patients.