Abstract:
An analytical method was developed for determining phospholipids in egg by high-performance liquid chromatography with an evaporative light scattering detector (HPLC-ELSD). Yolk phosphatidylcholine was successfully separated on a Intersil SIL-100A column(4.6 mm×100 mm, 5 μm), using methanol-acetonitrile-isopropanol as mobile phase at a flow rate of 0.8 mL/min and column temperature was set at 30 ℃. Air was used as atomizing gas and the gas flow rate was 1.7 L / min and the drift tube temperature was 45 ℃. The results showed that the linear range of phosphatidylcholine and phosphatidylethanolamine were 0.052~1.031 mg/mL and 0.041~0.823 mg/mL. The relative standard deviations(RSDs) of phosphatidylcholine and phosphatidylethanolamine in egg were 2.55% and 2.50%, the recovery rates were 93.0%~101.8% and 96.7%~106.7%, indicating high reproducibility and accuracy. The results of the determination by known content egg showed that the results by the evaporative light detector were more consistent with the true value than the ultraviolet detector. Therefore, this developed method can be applied to more accurate determine of phosphatidylcholine in egg and it is an ideal method.