Neutrophil-mediated phospholipid peroxidation assessed by gas chromatography-mass spectroscopy.
Disease pathophysiology frequently involves manifestations of the systemic inflammatory response syndrome. Oxyradicals represent key inflammatory mediators, and neutrophils are one important source of oxyradicals. This investigation examined neutrophil-mediated peroxidation of dilinoleoyl phosphatidyl-choline (DLPC) liposomes by monitoring the appearance of monohydroxyl linoleic acid with the use of gas chromatography-mass spectroscopy (GC-MS), compared with traditional assessment of thiobarbituric acid-reactive species (TBARS) and phosphatidylcholine-specific conjugated dienes. DLPC was peroxidized in a system using activated neutrophils in balanced salt solution containing chelated iron. 9-Monohydroxyl linoleic acid and 13-monohydroxyl linoleic acid were readily identified in neutrophil-mediated peroxidized DLPC with the use of GC-MS. Neutrophil NADPH oxidoreductase specific activity correlated highly with total ion current or specific ion monitoring of integrated peak areas for peroxidized linoleic acid but correlated poorly with DLPC-derived TBARS or conjugated dienes. These results ascertain that activated neutrophils mediate phosphatidylcholine lipid peroxidation to specific products, which may be precisely monitored with the use of GC-MS. The extent of this peroxidation is highly correlated with the magnitude of the neutrophil respiratory burst.
Zimmerman
J J
JJ
Department of Pediatrics, University of Wisconsin Children's Hospital, Madison 53792-4108, USA.
Ciesielski
W
W
Lewandoski
J
J
D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D008027:Light; D009504:Neutrophils / Q000502:physiology*; D010084:Oxidation-Reduction; D010545:Peroxides / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D010545:Peroxides; D010713:Phosphatidylcholines; D010743:Phospholipids; D017392:Thiobarbituric Acid Reactive Substances; C002448:1,2-linoleoylphosphatidylcholine