Critical assessment of three high performance liquid chromatography analytical methods for food carotenoid quantification |
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Authors: | M Graça Dias Luísa Oliveira M Filomena GFC Camões Baltazar Nunes Pieter Versloot Paul JM Hulshof |
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Institution: | 1. Departamento de Alimentação e Nutrição, Instituto Nacional de Saúde Doutor Ricardo Jorge (INSA, I.P.), Av. Padre Cruz, 1649-016 Lisboa, Portugal;2. CCMM/DQB – Faculdade de Ciências da Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal;3. Departamento de Epidemiologia, Instituto Nacional de Saúde Doutor Ricardo Jorge (INSA, I.P.), Av. Padre Cruz, 1649-016 Lisboa, Portugal;4. Division of Human Nutrition, Wageningen University, P.O. Box 8129, 6700 EV, Wageningen, The Netherlands |
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Abstract: | Three sets of extraction/saponification/HPLC conditions for food carotenoid quantification were technically and economically compared. Samples were analysed for carotenoids α-carotene, β-carotene, β-cryptoxanthin, lutein, lycopene, and zeaxanthin. All methods demonstrated good performance in the analysis of a composite food standard reference material for the analytes they are applicable to. Methods using two serial connected C18 columns and a mobile phase based on acetonitrile, achieved a better carotenoid separation than the method using a mobile phase based on methanol and one C18-column. Carotenoids from leafy green vegetable matrices appeared to be better extracted with a mixture of methanol and tetrahydrofuran than with tetrahydrofuran alone. Costs of carotenoid determination in foods were lower for the method with mobile phase based on methanol. However for some food matrices and in the case of E–Z isomer separations, this was not technically satisfactory. Food extraction with methanol and tetrahydrofuran with direct evaporation of these solvents, and saponification (when needed) using pyrogallol as antioxidant, combined with a HPLC system using a slight gradient mobile phase based on acetonitrile and a stationary phase composed by two serial connected C18 columns was the most technically and economically favourable method. |
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Keywords: | Carotenoids Food HPLC Method development Solvent and time reduction |
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