Abstract:
Quantification of malondialdehyde (MDA) as a
marker of lipid peroxidation is relevant for many research
fields. We describe a new sensitive and selective method to
measure free and total plasmatic MDA using derivatization
with 2,4-dinitrophenylhydrazine (DNPH) and ultra-HPLChigh-
resolution MS. Free and total MDA were extracted from
minute sample amounts (10 l) using acidic precipitation
and alkaline hydrolysis followed by acidic precipitation, respectively.
Derivatization was completed within 10 min at
room temperature, and the excess DNPH discarded by liquidliquid
extraction. Quantification was achieved by internal
standardization using dideuterated MDA as internal standard.
The method’s lowest limit of quantification was 100 nM
and linearity spanned greater than three orders of magnitude.
Intra- and inter-day precisions for total MDA were 2.9%
and 3.0%, respectively, and those for free MDA were 12.8%
and 24.9%, respectively. Accuracy was 101% and 107% at low
and high concentrations, respectively. In human plasma, free
MDA levels were 120 nM (SD 36.26) and total MDA levels
were 6.7 M (SD 0.46). In addition, we show the applicability
of this method to measure MDA plasma levels from a
variety of animal species, making it invaluable to scientists in
various fields. —Mendonça, R., O. Gning, C. Di Cesaré, L.
Lachat, N. C. Bennett, F. Helfenstein, and G. Glauser. Sensitive
and selective quantification of free and total malondialdehyde
in plasma using UHPLC-HRMS. J. Lipid Res. 2017.
58: 000–000.
