The carotenoid composition of Brazilian Valencia orange juice was determined by open column chromatography (OCC) and high-performance liquid chromatography. Carotenoid pigments were extracted using acetone and saponified using 10% methanolic potassium hydroxide. Sixteen pigments were isolated by OCC and identified as a-carotene, f-carotene, b-carotene, a-cryptoxanthin, b-cryptoxanthin, lutein-5,6-epoxide, violaxanthin, lutein, antheraxanthin, zeaxanthin, luteoxanthin A, luteoxanthin B, mutatoxanthin A, mutatoxanthin B, auroxanthin B and trollichrome B. Thirteen carotenoid pigments were separated using a ternary gradient (acetonitrile–methanol–ethyl acetate) elution on a C18 reversed-phase column. Among these, violaxanthin, lutein, zeaxanthin, b-cryptoxanthin, f-carotene, a-carotene, and b-carotene were quantified. The total carotenoid content was 12 ± 6.7 mg/l, and the major carotenoids were lutein (23%), b-cryptoxanthin (21%), and zeaxanthin (20%).