Ultra-Performance Liquid Chromatographic Determination of Tocopherols and Retinol in Human Plasma

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...

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Bibliographic Details
Published in:Journal of chromatographic science 2014-10, Vol.52 (9), p.1065-1070
Main Authors: Bell, Edward C., John, Mathew, Hughes, Rodney J., Pham, Thu
Format: Article
Language:English
Subjects:
Acetates
Assaying
Chromatography
Chromatography, High Pressure Liquid - instrumentation
Chromatography, High Pressure Liquid - methods
Chromatography, High Pressure Liquid - standards
Chromatography, Reverse-Phase - methods
Human
Humans
Ingestion
Limit of Detection
Liquid Chromatography
Methyl alcohol
Quality Control
Tocopherol
Tocopherols - blood
Vitamin A - analogs & derivatives
Vitamin A - blood
Vitamin A - chemistry
Vitamins
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Summary: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.
ISSN:0021-9665
1945-239X
DOI:10.1093/chromsci/bmt161