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Improved method for plasma malondialdehyde measurement by high-performance liquid chromatography using methyl malondialdehyde as an internal standard
Measurement of malondialdehyde (MDA) is an important
contribution to the assessment of oxidative stress. We report a
sensitive and reproducible high-performance liquid chromatography
(HPLC) method for measurement of plasma MDA in the assessment of lipid
peroxidation. Using methyl malondialdehyde (Me-MDA) as an internal
standard with reversed-phase HPLC and UV detection and derivatisation
with 2,4 dinitrophenylhydrazine (DNPH), we obtained maximum MDA values
with 60-min incubation of 10% plasma with 1 M NaOH at 60 degreesC. The
dilution of the plasma and a longer incubation time in the alkaline
hydrolysis step greatly improved recovery of MDA from its bound form.
Ratios of peak height of MDA/Me-MDA were linear over a range of 0-100
muM with correlation coefficients >0.99. The recovery was 88.5%.
Within and between run variations were <4 and <7%, respectively.
The mean MDA value measured in 20 healthy volunteers was 13.8 muM
(+/-1.32). (C) 2002 Elsevier Science B.V. All rights reserved
