Comparison of 3 Spectrophotometric Methods for Carotenoid Determination in Frequently Consumed Fruits and Vegetables

Carotenoids are C-40 tetraterpenoid compounds with potential health beneficial effects. Major dietary sources include a variety of fruits and vegetables. Rapid screening methods are therefore desired, but their accuracy varies depending on the carotenoid profile and the matrix of the plant food. In the present study, 3 different methods were compared, all based on a rapid extraction protocol and spectrophotometric measurements to determine the total amount carotenoids present in fruits and vegetables (n = 28), either with or without chlorophyll. Published methods (a) Lichtenthaler and (b) Hornero-Méndez and Mínguez-Mosquera were compared with a newly developed method (method c) based on the average molar absorption coefficient (135310 Lcm⁻¹mol⁻¹) and wavelength (450 nm in acetone), for the 5 predominant carotenoid species (beta-carotene, zeaxanthin, lycopene, lutein, beta-cryptoxanthin) in the investigated foods. All results were compared to HPLC (method d). To avoid overestimating carotenoid concentrations due to chlorophyll A and B presence, the effect of saponification was studied for all methods. Overall, saponification led to significant carotenoid losses (12.6 ± 0.9%). Methods a, b, c, and d yielded 5.1 ± 0.4 mg/100 g, 4.6 ± 0.5 mg/100 g, 4.3 ± 0.5 mg/100 g, and 4.2 ± 0.5 mg/100 g total carotenoids, respectively, with method a leading to significant higher mean concentrations compared to all other methods (P < 0.001, Bonferroni) with methods b and c being not significantly different and highly correlated compared to HPLC (> r = 0.95). Similar results were found when stratifying for chlorophyll content and fruits compared with vegetables, however, accuracy varied for individual fruits, highlighting the limitation to use the same method for all plant foods. Practical Application: This study presents a comparison of various rapid spectrophotometric measurements to determine total carotenoid content in various fruits and vegetables and could aid in the selection of the appropriate method for individual plant foods with different carotenoid profile and matrices.