Oxidatively modified plasma phospholipids containing reactive carbonyl functions measured by HPLC: evidence for phosphatidylcholine-bound aldehydes in plasma of burn patients

Free Radic Res. 1995 Apr;22(4):327-36. doi: 10.3109/10715769509145645.

Abstract

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.

MeSH terms

  • Adult
  • Aldehydes / blood*
  • Burns / metabolism*
  • Chromatography, High Pressure Liquid
  • Edetic Acid / metabolism
  • Female
  • Humans
  • Hydrogen Peroxide / metabolism
  • Iron Chelating Agents / metabolism
  • Male
  • Middle Aged
  • Oxidation-Reduction
  • Phenylhydrazines / metabolism
  • Phosphatidylcholines / blood*
  • Phosphatidylcholines / metabolism
  • Soybeans
  • Spectrophotometry

Substances

  • Aldehydes
  • Iron Chelating Agents
  • Phenylhydrazines
  • Phosphatidylcholines
  • 2,4-dinitrophenylhydrazine
  • Edetic Acid
  • Hydrogen Peroxide