Efficient HPLC Separation on a Core-C30 Column with MS2 Characterization of Isomers, Derivatives and Unusual Carotenoids from Tomato Products

Abstract

An efficient and rapid liquid chromatographic method was developed for the separation of carotenoids and their geometrical isomers in tomato products using a core C30 column of 2.6 μm particles with gradient elution of tert-butyl-methyl-ether in 2% water in methanol. Excellent separation of the major carotenoids such as lycopene, β-carotene and lutein as well as their geometrical isomers and oxygen-containing derivatives with resolution factors ranging between 0.78 and 4.0 and selectivity of 1.01-1.63 was achieved. Validation of the developed method met the acceptance criteria concerning linearity, recovery, precision and limit of detection and quantification. Calibrations were linear with correlation coefficient (R2) values between 0.9966 and 0.9999. The limit of detection and quantification values were found to be 0.008 and 0.017 and 0.029 and 0.056 μg/mL, respectively. Recovery of 94.3-99.9%, intraday precision of 1.81-4.45% and interday precision of 3.13-6.86% were obtained. The hyphenation of liquid chromatography with diode-array and mass spectrometry was helpful in the identification of the separated carotenoids particularly the unusual di-hydroxy cyclolycopene adduct and di-methoxy lycopene determined for the first time in tomato products. Commercially available kinds of tomato juice and ketchup were evaluated based on their carotenoid content.