Applications on the monitoring of oxidative modification of LDL by capillary electrophoresis: a comparison with spectrophotometer assay.
The aim of this work is the application of Stocks and Miller capillary electrophoresis (CE) method in order to evaluate the human LDL susceptibility to Cu(2+)-induced oxidation. Lipid peroxidation determines a change in the relative electrophoretic mobility (REM) of lipoprotein that can be monitored by capillary electrophoresis using uncoated fused silica capillaries and tricine-methylglucamine as electrophoretic buffer. We have evaluated the differences in the susceptibility to oxidation of LDL subjected to different preparations (dialysis or gel filtration, after ultracentrifugation, to remove EDTA), and different storage (4 degrees C for 1 week or lyophilization) by measuring REM and lipid hydroperoxides (ROOH) with a spectrophotometer assay. Our results indicate that gel filtration is a more convenient procedure than dialysis for the isolation of LDL and that lyophilised samples are less prone to oxidation than those stored at 4 degrees C. Moreover, REM appears to be a more sensitive and easier method than spectrophotometer assay both to monitor the oxidative modification of LDL and to evaluate oxidative state of native LDL.
Zinellu
Angelo
A
Chair of Clinical Biochemistry, University of Sassari, Sassari, Italy.
Sotgia
Salvatore
S
Galistu
Franca
F
Lumbau
Fiorenza
F
Pasciu
Valeria
V
Mario Pes
Giovanni
G
Tadolini
Bruna
B
Deiana
Luca
L
Carru
Ciriaco
C