Phosphatidylcholine isolation from egg yolk phospholipids by high-performance liquid chromatography

Cell membrane components have been increasingly recognized as important biochemicals in the fields of biochemistry and pharmacy due to their relationship with metabolite transport in the cells. Among the components, phosphatidylcholine (PC) is considered a valuable biochemical, because it is difficult to commercialize. PC demand has been largely increased in the fields of the nutrient, cosmetic and pharmacy industries, and so the development of a preparative chromatography process is critical to supply a low-cost PC. In this study, we investigated the HPLC separation of phospholipid originated from egg yolk, which contains 80% (w/w) PC and 15% (w/w) phosphatidylethanolamine. Column temperature, mobile phase composition and its flow-rate and kinds of stationary phase were varied to understand the effectiveness of PC separation. For studying the relationship between recovery yield and sample loading amount in HPLC, we performed overloading experiments. In this way, we successfully separated PC with over 99% purity and with 98% yield with the following HPLC operating conditions; pure methanol as a mobile phase, 2.0 ml/min flow-rate and 1000 mg/ml feed concentration in a KR-100-10SIL column.