This study developed a method for simultaneously determining 10 carotenoids of lutein, zeaxanthin, α-cryptoxanthin, β-cryptoxanthin, ε-carotene, α-carotene, β-carotene, （6R）-δ-carotene, γ-carotene, and lycopene by reverse-phase C30 high performance liquid chromatography. Carotenoid extracts for each sample were separated by a reverse-phase C30 column（YMC carotenoid C30, 250 mm×4.5 mm, 5 μm）with its column temperature of（25±1）℃, and eluted using the mobile phase A（A1∶B1=9∶1, V/V）and B（A1∶B1=1∶9, V/V）with the flow rate of 1 mL/min. The mobile phase A1 consisted of methanol：water（97∶3, V/V）with 0.05 mol/L ammonium acetate and 0.1%（W/V）2, 6-di-tert-butyl-4-methylphenol（BHT）, and the mobile phase B1 consisted of 100% methyl tert-butyl ether with 0.1%（W/V）BHT. The effluents were then monitored at 450 nm using a photo diode array detector. The calibration curves of these ten carotenoids showed a good linearity with the concentration range of 0.5-20 μg/mL and linear relative coefficients（R2）were > 0.995；limit of detection（S/N=3）was 0.01-1.6 μg/mL, limit of quantitation（S/N=10）was 0.2-4.0 μg/mL, average recovery was 96.29%-104.47%, relative standard deviation was of 0.03%-1.24%, with 40 min of whole process. The method has been applied for measuring carotenoids content in banana fruit and maize kernel samples, i.e., verifying the stability, accuracy and reliability of this method in real agricultural biological samples.
