Separation and determination of in vitro oxidized phospholipids by capillary zone electrophoresis

A simple capillary zone electrophoresis (CZE) method was used to determine in vitro oxidized phosphatidyl choline (ox-PC). To optimize the capillary electrophoresis (CE) conditions, organic buffer additives, buffer ionic strength, buffer pH, and applied voltage were examined. The optimal CE separation buffer chosen was an aqueous–organic solvent system containing 10% sodium phosphate buffer (5 mM, pH 7.40), 80% methanol, and 10% acetonitrile. One major peak with a small shoulder was found for phosphatidyl choline (PC), whereas one major peak and a complex region containing several lower-mobility peaks were found for ox-PC. The lower-mobility species of ox-PC has high levels of conjugated dienes characterized by strong absorbance at 234 nm. The electropherograms of PC and ox-PC were significantly different and highly reproducible. The intensities of lower-mobility species decreased significantly when the antioxidant vitamin C concentration was increased from 6 to 600 μM. This study provides a simple CZE method to differentiate in vitro oxidized from nonoxidized PC molecular species.