Human plasma concentrations of malondialdehyde (MDA) and the F2-isoprostane 15(S)-8-iso-PGF(2alpha) may be markedly compromised by hemolysis: evidence by GC-MS/MS and potential analytical and biological ramifications.
OBJECTIVE
Malondialdehyde (MDA) and the F(2)-isoprostane 15(S)-8-iso-prostaglandin F(2alpha) (15(S)-8-iso-PGF(2alpha)) belong to the most frequently analyzed biomarkers of oxidative stress in basic and clinical research. The objective of the present study was to examine the effect of hemolysis on free MDA and total (free+esterified) 15(S)-8-iso-PGF(2alpha) concentrations in human plasma.


METHODS
MDA and 15(S)-8-iso-PGF(2alpha) were determined by GC-MS/MS in plasma samples from venous heparinized blood drawn under resting conditions (n=22) as well as under physical exercise (n=158) in 22 healthy young subjects. In vitro, we prepared plasma samples with hemolysis degrees up to 0.8% using artificially hemolyzed, freshly obtained heparinized blood.


RESULTS
In some plasma samples of the exercise study both under resting and exercise conditions, clinically significant hemolysis was macroscopically visible. Both in vivo (r=0.74) and in vitro (r=0.87), we found a significant positive correlation between hemolysis degree (0-0.2%) and MDA plasma concentrations (50-250 nmol/L). Unlike in vitro (r=0.84), in vivo, 15(S)-8-iso-PGF(2alpha) and MDA plasma concentrations correlated weakly (r=0.50).


CONCLUSIONS
We hypothesize that free hemoglobin catalyzes the formation of MDA and 15(S)-8-iso-PGF(2alpha) from free and esterified arachidonic acid. Plasma concentrations of MDA and total 15(S)-8-iso-PGF(2alpha) may be markedly compromised by hemolysis. Measurements of MDA and 15(S)-8-iso-PGF(2alpha) should be treated with caution regarding involvement of oxidative stress in disease as well as in health both under resting conditions and under exercise.
Dreissigacker
Ulrike
U
Institute of Sport Medicine, Hannover Medical School, Carl-Neuberg-Str 1, 30625 Hannover, Germany.
Suchy
Maria-Theresia
MT
Maassen
Norbert
N
Tsikas
Dimitrios
D
D000328:Adult; D016718:Arachidonic Acid / Q000378:metabolism; D015415:Biomarkers / Q000097:blood / Q000737:chemistry; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000097:blood; D028441:F2-Isoprostanes / Q000097:blood / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D006461:Hemolysis; D006801:Humans; D008297:Male; D008315:Malondialdehyde / Q000097:blood* / Q000737:chemistry; D015394:Molecular Structure; D018384:Oxidative Stress; D014662:Vasoconstrictor Agents / Q000097:blood* / Q000737:chemistry
D015415:Biomarkers; D028441:F2-Isoprostanes; D014662:Vasoconstrictor Agents; C075750:8-epi-prostaglandin F2alpha; D016718:Arachidonic Acid; D008315:Malondialdehyde; D015237:Dinoprost