Abstract: A HPLC method for simultaneous determination of astaxanthin, lutein, cantharidin, zeaxanthin and β-carotene in eggs and chicken was developed. The results showed a good linear relationship in the range of 0.5~100 μg/mL for five carotenes, and the coefficients of determination were all above 0.996. The limits of detection of the method were between 0.10 and 0.20 mg/kg, with the limits of quantitation were between 0.33 and 0.66 mg/kg. At the spiked levels of 1, 10 and 20 mg/kg, the recoveries of eggs and chicken samples were 71.6%~112.9% and 88.6%~119.7%, respectively, with RSD≤7.2%. The method is rapid, simple, accurate and reproducible. It is suitable for the detection of five kinds of carotenoids in complex matrix samples such as eggs and chicken. It is of great significance for the market supervision of eggs and chicken.