Quantification of Vitamins A, E, and K and Carotenoids in Submilliliter Volumes of Human Milk

Journal of AOAC International
Antoine LévèquesEsther Campos-Giménez

Abstract

Background: Human milk is the optimal nutrition for all newborns in the first 6 months of life. In order to assess the nutritional needs of the breastfed infant, human milk is often characterized for multiple nutrients. Objective: To ensure that we minimize the volume of milk dedicated for research and optimize the number of nutrients characterized, we developed analytical methodologies for the determination of vitamins A (retinol), E (alpha and gamma tocopherol), K (phylloquinone and menaquinone-4), and five carotenoids (β-carotene, lycopene, β-cryptoxanthin, lutein, and zeaxanthin) using <1 mL human milk. Method: Vitamins E and K and carotenoids are simultaneously isolated from 750 μL milk by liquid-liquid extraction (LLE). Tocopherols and carotenoids are determined by normal-phase LC with fluorescence and ultraviolet detection respectively. Vitamin K is analyzed on the same extracts after resuspension and clean-up by reversed phase liquid chromatography coupled to tandem MS. The analysis of vitamin A involves saponification of 200 μL milk followed by LLE and determination by normal-phase LC with UV detection. Results: Full single-laboratory validation at four different concentration levels is presented. Recovery rates were w...Continue Reading

Citations

Apr 2, 2021·Critical Reviews in Food Science and Nutrition·Jimena RíosSamanta Thomas-Valdés
Sep 3, 2021·Nutrition Reviews·Přemysl MladěnkaUNKNOWN OEMONOM researchers and collaborators

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