Radiation-induced peroxidation of small unilamellar vesicles of phosphatidylcholine generated by sonication.
The present study was aimed at determining the peroxidation of model membranes constituted of liposomes of 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (PLPC) submitted to hydroxyl free radicals (generated by gamma-radiolysis) attack. Liposomes of PLPC were prepared using the sonication technique, and dynamic light-scattering (DLS) measurements allowed characterization of the liposomal dispersions. Irradiation damages in sonication-generated liposomes were assessed by monitoring several oxidation products, such as conjugated dienes (by means of UV--visible spectrophotometry) and hydroperoxides (using reverse phase high-performance liquid chromatography (HPLC) associated with chemiluminescence detection). It has been shown that three different families of hydroperoxides are formed: the first one (at low radiation doses) results from HO. attack on the linoleyl chain of PLPC, giving phosphatidylcholine hydroperoxides possessing a conjugated dienic structure; the two others (at high radiation doses) are obtained by the secondary HO. attack on the primary hydroperoxide family. The quantification of these products associated with the comparison of their radiation-dose-dependent formation has provided valuable information concerning the mechanisms of their formation. Analysis by HPLC -- mass spectrometry has confirmed the presence of hydroperoxides and underlined various other products, like chain-shortened fragments and oxygenated derivatives of polyunsaturated sn-2 fatty acyl chain residues. Structural assignment proposals of some oxidation products have been proposed.
Vitrac
H
H
Laboratoire de Chimie Physique, UMR 8601 -- CNRS, Université Paris 5, 45 rue des Saints-Pères, 75270 Paris CEDEX 06, France. Heidi.Vitrac@univ-paris5.fr
Courrègelongue
M
M
Couturier
M
M
Collin
F
F
Thérond
P
P
Rémita
S
S
Peretti
P
P
Jore
D
D
Gardès-Albert
M
M
D002851:Chromatography, High Pressure Liquid; D015227:Lipid Peroxidation; D008081:Liposomes; D010316:Particle Size; D010713:Phosphatidylcholines / Q000528:radiation effects*; D013010:Sonication
D008081:Liposomes; D010713:Phosphatidylcholines; C055402:1-palmitoyl-2-linoleoylphosphatidylcholine