Oxidatively modified plasma phospholipids containing reactive carbonyl functions measured by HPLC: evidence for phosphatidylcholine-bound aldehydes in plasma of burn patients.
A HPLC method has been developed to measure phosphatidylcholine (PC) containing reactive carbonyl functions in the sn-acyl residue in order to study processes in which such reactive carbonyls can be formed due to e.g. oxidative fragmentation. The method has been applied to determine PC-bound carbonyls as 2,4-dinitrophenylhydrazones (DNPH) in plasma of burn patients. Plasma from healthy volunteers served as controls. Additionally, in vitro oxidation experiments (A: plasma, buffer diluted; B: plasma + iron-EDTA complex and C: plasma + iron-EDTA complex + H2O2) have been performed to obtain and to identify 2,4-dinitrophenylhydrazine derivatizable carbonyl functions in plasma PC. Both, the PC-aldehydes and PC-aldehyde DNPH derivatives were cleavable with phospholipase C. Quantification was based on thin-layer chromatography purified soybean phosphatidylcholine, which was identically oxidized and derivatized as the plasma lipids in vitro.
Gasser
H
H
Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria.
Hallström
S
S
Redl
H
H
Schlag
G
G
D000328:Adult; D000447:Aldehydes / Q000097:blood*; D002056:Burns / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004492:Edetic Acid / Q000378:metabolism; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007502:Iron Chelating Agents / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D010084:Oxidation-Reduction; D010659:Phenylhydrazines / Q000378:metabolism; D010713:Phosphatidylcholines / Q000097:blood* / Q000378:metabolism; D013025:Glycine max; D013053:Spectrophotometry
D000447:Aldehydes; D007502:Iron Chelating Agents; D010659:Phenylhydrazines; D010713:Phosphatidylcholines; C004787:2,4-dinitrophenylhydrazine; D004492:Edetic Acid; D006861:Hydrogen Peroxide