title: Free and bound malondialdehyde measured as thiobarbituric acid adduct by HPLC in serum and plasma.
abstract: 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.
authors.affiliation: Laboratoire de Biochimie Médicale A., Université de Bordeaux II, France.
authors.forename: M A
authors.initials: MA
authors.lastname: Carbonneau
authors.forename: E
authors.initials: E
authors.lastname: Peuchant
authors.forename: D
authors.initials: D
authors.lastname: Sess
authors.forename: P
authors.initials: P
authors.lastname: Canioni
authors.forename: M
authors.initials: M
authors.lastname: Clerc
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D005260:Female; D006801:Humans; D015227:Lipid Peroxidation; D008297:Male; D008315:Malondialdehyde / Q000097:blood*; D008875:Middle Aged; D009369:Neoplasms / Q000097:blood; D006435:Renal Dialysis; D013858:Thiobarbiturates / Q000097:blood* / Q000737:chemistry
chemical_list: D013858:Thiobarbiturates; D008315:Malondialdehyde; C029684:thiobarbituric acid