HPLC analysis of carotenoids from tomatoes using cross-linked C18 column and MS detection

This study was conducted to analyze carotenoids from tomatoes by high-performance liquid chromatography using reversed-phase C18 silica having cross-linked end-capping with diode array and mass spectrometric detection. An efficient gradient elution system was developed to achieve good and reliable separation of both major and minor carotenoids as well as their isomers. Resolution of lycopene, β-carotene and their isomers was 0.91-3.97 and 1.02-2.86 with cross-linked and conventional C18 column, respectively. The % recovery for zeaxanthin, lycopene and β-carotene was found to be in the range of 89-97%. Limits of detection and quantification of 19.44 and 64.79 ng/mL for zeaxanthin, 15.6 and 52.4 ng/mL for lycopene and 8.28 and 27.61 ng/mL for β-carotene were determined. More carotenoid compounds could be separated and detected with the new method as compared with conventional C18 column. Hyphenation of HPLC with photodiode array and mass spectrometry detectors assisted in detection of tetra-dehydrocarotenoid and fatty acid diesters of xanthophylls in tomato products. Content of all-trans-lycopene, β-carotene and total carotenoid in different industrial tomatoes tested was found to range between 41.87 and 84.65, 0.89 and 1.50 and 53.22 and 112.60 µg/g fresh weight, respectively.