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 degrees C. 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 microM 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 microM (+/-1.32).
Sim
Ah Siew
AS
Cardiovascular Genetics Laboratory, Department of Medicine, Prince of Wales Hospital, High Street, Randwick, Sydney, NSW Australia 2031.
Salonikas
Chris
C
Naidoo
Daya
D
Wilcken
David Emil Leon
DE
D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006834:Hydrazines; D006868:Hydrolysis; D007202:Indicators and Reagents; D015227:Lipid Peroxidation; D008315:Malondialdehyde / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry*; D013058:Mass Spectrometry; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis
D006834:Hydrazines; D007202:Indicators and Reagents; D017392:Thiobarbituric Acid Reactive Substances; C446799:dinitrophenylhydrazine; C033319:methylmalondialdehyde; D008315:Malondialdehyde