An isocratic high performance liquid chromatographic (HPLC) method with run times of less than 22 min has been developed for simultaneous analysis of carotenoids and tocopherols in corn. The reverse phase system utilizes a nonaqueous mobile phase in combination with a polymeric C18column coupled to a monomeric C18column to separate lutein, zeaxanthin, β-cryptoxanthin, α-carotene, β-carotene, and the α-, δ- and γ- forms of tocopherol. The detector was a multi-channel UV-VIS. One channel was used to monitor carotenoid absorption at 450 nm while another monitored tocopherols at 290 nm.