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 microM. This study provides a simple CZE method to differentiate in vitro oxidized from nonoxidized PC molecular species.
Ho
Yu-Ling
YL
Department of Chemistry, National Changhua University of Education, Changhua 50058, Taiwan.
Chiu
Jing-Huei
JH
Wu
Chung-Yu
CY
Liu
Mine-Yine
MY
D000975:Antioxidants / Q000737:chemistry; D001205:Ascorbic Acid / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D010084:Oxidation-Reduction; D010713:Phosphatidylcholines / Q000302:isolation & purification*; D010743:Phospholipids / Q000032:analysis / Q000302:isolation & purification*
D000975:Antioxidants; D010713:Phosphatidylcholines; D010743:Phospholipids; D001205:Ascorbic Acid