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- Title
Ultra-Performance Liquid Chromatographic Determination of Tocopherols and Retinol in Human Plasma.
- Authors
Bell, Edward C.; John, Mathew; Hughes, Rodney J.; Pham, Thu
- Abstract
A rapid, selective and sensitive ultra-performance liquid chromatography method has been developed for the detection and quantification of tocopherols and retinol in human plasma. Alpha-tocopherol, gamma-tocopherol and retinol are assayed using fluorescence detection. Excitation/emission wavelengths are 295/330 nm and 325/470 nm for the analysis of both tocopherols and retinol, respectively. Retinol acetate is employed as the internal standard. The reversed-phase method incorporates gradient elution with a mobile phase consisting of methanol and acetonitrile. Separation of vitamin compounds is achieved using a bridged ethyl hybrid C18 column. The retention times for retinol, retinol acetate, gamma-tocopherol and alpha-tocopherol are 1.6, 1.8, 3.9 and 4.3 min, respectively. The limits of quantification for retinol, gamma-tocopherol and alpha-tocopherol were 0.02, 0.02 and 0.1 µg/mL, respectively. The assay method is suitable for the analysis of tocopherols and retinol in human plasma. The method may be applied following the ingestion of foods fortified with these fat-soluble vitamins.
- Subjects
LIQUID chromatography; VITAMIN E; VITAMIN A; WAVELENGTHS; RF values (Chromatography)
- Publication
Journal of Chromatographic Science, 2014, Vol 52, Issue 9, p1065
- ISSN
0021-9665
- Publication type
Article
- DOI
10.1093/chromsci/bmt161