Calculation of the HPLC chromatographic retention times of different carotenoids supported our improved chromatographic separation of beta-carotene and lutein in four tomatoes and two tomatillo varieties in fresh form and after three different cooking procedures: pot boiling, cooking in a pressure cooker and microwaving. A good separation was achieved experimentally using an Ultrasphere ODS column and gradient elution with an acetonitrile-tetrahydrofuran-water mobile phase. It was shown that diverse tomato species contained different amounts of beta-carotene (6-400 microg/100 mg) and lutein (2-30 microg/100 mg). The concentration in fresh samples was higher than in cooked tomatoes. The beta-carotene content in fresh tomatillo varied between 2 and 20 microg/100 mg. Microwaving caused partial destruction of the beta-carotene and lutein in tomatillos.