Abstract: Objective: To determine the content of diethylstilbestrol and testosterone in HY-LINE Variety Brown chick embryo and egg. Methods: RP-HPLC method was used to determine the content of diethylstilbestrol and testosterone in chick embryo. The analysis was carried out on Waters XTerra C18 column (150 mm×4.6 mm, 5μm) with the mobile phase methanol-acetonitrile-water(2:2:1, V/V/V), flow rate 1.0 ml/min, temperature 40 ℃and wavelength 240 nm. Results and conclusion: The method is simple and has a good linear relationship in the range of 0.05～3.500 μg/ml. The linear equation of diethylstilbestrol is y = 21734x + 963.18 with correlation coefficient 0.9991, and the linear equation of testosterone is y = 28967x-475.62, with correlation coefficient 0.9995. The limit of detection of diethylstilbestrol is 0.25 ng, and that of testosterone is 0.33 ng. The average recovery ranges from 78.7% to 98.4%. The relative standard deviation is less than 2.1%.

Key words: diethylstilbestrol, testosterone proprionate, RP-HPLC, chick embryo