Tomatoes are one of the most popular vegetables due of their wide use as food. Tomatoes are not only tasty fruit, but one of its benefits-high carotenoids content is well-known. Non-destructive analyses methods are used more and more in different industries. It is cheaper, faster and environmentally friendly way of analyse than traditional chemical methods. But these methods need references to the traditional ones. The aim of this study was to find the correlation between lycopene and β-carotene content in tomatoes determined with reflectance spectrometer and extraction of pigments. Content of two carotenoids (lycopene and β-carotene) was determined in 27 varieties of tomatoes. Red, pink, orange, yellow and brown fruits were included in experiment. Reflectance spectrums of tomatoes fruits were obtained with remote sensing portable spectroradiometer RS-3500 (Ltd.Spectral Evolution). Tetrahydrofuran was used for extraction of pigments. Absorption spectra of extract were obtained by spectrophotometer UV-Vis-1800 (Ltd. Shimadzu). Linear regression analyses were performed to correlate spectral data with lycopene and β-carotene concentrations measured by pigment extraction. The best reflectance region for lycopene spectral detection was 570 ± 5 nm, but for β-carotene 487 ± 5 nm. Reflectance indexes for both pigments were worked out. High linear correlation (R2 > 0.9) between spectral parameters and lycopene concentration was detected. Correlation between results obtained with methods used for β-carotene determination was lower and depended of colour of tomatoes fruits.