HPLC Analysis of Egg Yolk Phosphatidylcholine by Evaporative Light Scattering Detector

Abstract

Egg yolk phosphatidylcholine (EYPC) is being widely used in food and pharmaceutical industries nowadays owing to its surface activity, pharmaceutical usefulness, and so on. Common determination methods of phospholipids were based on the American Oil Chemists' Society (AOCS) Official Method Ja7b-91, in which n-hexane/2-propanol/acetate buffer was used as the mobile phase. In order to achieve desired results, gradient elution or buffer solution was used, which made the detection process more complicated. Moreover, water or buffer solution could affect the silica gel column both on its lifespan and the separation efficiency significantly. In this study, different mobile phase and detector were used to simplify EYPC analyzing process instead of using water within the mobile phase. The optimized HPLC operating conditions are as follows: pure methanol as a mobile phase, flow rate of 1.0 ml·min−1, silica gel column (250 mm×4.6 mm, 5 μm, Inertsil GLTM), column temperature 30°C and low temperature evaporative light scattering detector (40°C, 0.35 MPa) as used. Under this optimal condition, the linear relative coefficient of the standard curve is 0.998 and the recovery was in the range of 96.83%-101.58% with a relative standard deviation of 1.79% (n=6).