Abstract: In this study,a method for the simultaneous analysis and detection of lutein,zeaxanthin and β-cryptoxanthin in food was developed using high-performance liquid chromatography(HPLC). The optimal pretreatment conditions were adding with 30 mL 0.1% 2,6-di-tertbutyl-4-methyl phenol(BHT)ethanol solution and 10 mL 60% potassium hydroxide solution for samples. Then the samples were saponificated in a water bath for 15 min at a constant temperature of 50 ℃ by thermostatic waterbath,followed by extraction using 100 mL solvent mixture of n-hexane,ethyl ether and cyclohexane(2:2:1). The results showed that this method was superior to the national standard and ultrasonic extraction,because it could be applied rapidly and the mild extraction of lutein provided an enhanced extraction effect. A millet and rice sample was used for methodological verification and the results showed that the linear relations for lutein,zeaxanthin and β-cryptoxanthin were good in the range of 0.2342~9.3689,0.3004~12.0167 and 0.3734~14.9371 μg/mL,respectively. Six parallel experiments were conducted and the relative standard deviations of lutein and zeaxanthin were 1.41% and 4.98%,respectively,and their spiked recoveries were between 85.6% and 98.8%,thus indicating that this method had sound repeatability and reliability characteristics. The establishment of this method can make up for the current deficiency of relevant standards. Furthermore,it provides solutions to various problems in practical work,as well as technical assistant for product development,food evaluation and supervision in the food production industry.