High performance liquid chromatographic separation of hydroperoxy- phospholipids and their corresponding hydroxy-phospholipid derivatives

This paper describes high performance liquid chromatography (HPLC) conditions suitable for separating hydroperoxy-phospholipids from hydroxy- phospholipids. 1-stearoyl-2-(13-hydroperoxy-cis-9, trans-11-octadecadienoyl)- L-3-phosphatidylcholine, 1, 2 di (13-hydroperoxy-cis-9, trans-11- octadecadienoyl)-L-3-phosphatidylcholine, 1-palmitoyl-2-(13-hydroperoxy-cis- 9, trans-11-octadecadienoyl)-L-3-phosphatidylethanolamine and their corresponding hydroxy-phospholipid derivatives were resolved by HPLC on an Ultracarb 5 ODS (20) column at 30°C. Each pair of these phospholipids required different conditions for optimal separation but all used a mobile phase of acetonitrile-methanol-water mixture containing 10 mM choline chloride. The separation described allows accurate measurement of phospholipid hydroperoxide peroxidase activities.