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AACC International Approved Methods -AACC Method 86-06.01. Analysis of Vitamins A and E by High-Performance Liquid Chromatography

AACC International Approved Methods


AACCI Method 86-06.01
Analysis of Vitamins A and E by High-Performance Liquid Chromatography


This primary method measures the presence and quantity of vitamin A as retinol whether it is present as retinol or as retinol esters (retinyl acetate and/or retinyl palmitate) and vitamin E as alpha-tocopherol (expressed as alpha-tocopherol acetate) whether it is present as alpha-tocopherol or as alpha-tocopherol esters (such as alpha-tocopherol acetate) in foods. This method is applicable to all foods. The detection/quantitation limits are 0.15 µg/g for retinol and 0.001 mg/g for alpha-tocopherol. This method covers the range of 0.15 µg/g to 100% vitamin A and 0.001 mg/g to 100% vitamin E. Standards and samples are saponified in basic ethanol-water solution, neutralized, and diluted, converting fats to fatty acids and retinol esters and tocopherol esters to retinol and tocopherol, respectively. Retinol and alpha-tocopherol are quantitated on separate high-performance liquid chromatography (HPLC) systems, using UV detection at 313 or 328 nm for retinol and fluorescence detection (excitation 290 nm, emission 330 nm) for alpha-tocopherol. Vitamin concentrations are calculated by comparison of the peak heights or peak areas of vitamins in samples with those of standards. Caution: see Note 1.