An improved method of cholesterol determination in egg yolk by HPLC

An improved method for cholesterol determination in egg yolk is reported in this paper. Egg yolk was first diluted. Cholesterol was then extracted with ether and petroleum ether, and quantified by reverse phase chromatography on a Zorbax ODS column (0.46 x 15cm, 5-6 mu m) using a mobile phase of acetonitrile and 2-propanol (4:1) with a flow rate of 0.6 mL/min. A linear correlation was observed between cholesterol concentration at 0.05-0.40 mg/mL and its peak heights with a correlation coefficient of 0.9993 (n = 5). The average recovery was 98.9%, and detection limit was 0.02 mg/mL. No differences in cholesterol concentration were observed between egg yolk samples with and without saponification. Rapid and reproducible quantification of cholesterol in egg yolk can be completed with this simplified method.