What question did this study set out to answer?

To assign accurate values for vitamin D and its metabolites in food-matrix standard reference materials (SRMs).

April 7, 2026Journal of Agricultural and Food Chemistry2 citationsOpen Access

Value Assignment of Vitamin D and 25-Hydroxyvitamin D in Food-Matrix Standard Reference Materials (SRMs) Using Isotope Dilution Liquid Chromatography–Tandem Mass Spectrometry (ID LC–MS/MS)

CBCarolyn Q BurdetteNational Institute of Standards and Technology JYJames H. YenNational Institute of Standards and Technology AKAdam J. KuszakNational Institutes of Health

Key Points

To assign accurate values for vitamin D and its metabolites in food-matrix standard reference materials (SRMs).
Used isotope dilution liquid chromatography-tandem mass spectrometry (ID LC-MS/MS) for analysis.
Value assignment conducted for vitamin D2, vitamin D3, and 25-hydroxyvitamin D3 in various SRMs.
Analyzed five specific SRMs: Meat Homogenate, Whole Milk Powder, Bovine Liver, Whole Egg Powder, and Soy Milk.
Mass fractions for vitamin D3 ranged from 0.498 μg/kg to 49.5 μg/kg.
Mass fractions for 25-hydroxyvitamin D3 ranged from 0.53 μg/kg to 12.1 μg/kg.
These SRMs facilitate improved validation of analytical methods and comparability of measurements across laboratories.

Abstract

Discrepancies exist between reported vitamin D dietary intake and measured status. Hydroxylated metabolites such as 25-hydroxyvitamin D 25(OH)D may be more bioavailable; however, information on 25(OH)D content in foods is limited. To address this gap and support analytical method validation, NIST value assigned vitamin D2, vitamin D3, and 25-hydroxyvitamin D3 25(OH)D3 in five food-matrix Standard Reference Materials® (SRMs). Using isotope dilution LC-MS/MS, values were established for SRMs 1546a (Meat Homogenate), 1549a (Whole Milk Powder), 1577c (Bovine Liver), 1845a (Whole Egg Powder), and 3235 (Soy Milk). Mass fractions ranged from 0.498 μg/kg to 49.5 μg/kg for vitamin D3 and 0.53 μg/kg to 12.1 μg/kg for 25(OH)D3. These SRMs enable validation of analytical methods and measurement comparability assessment across laboratories. By providing accurate reference values for both parent vitamins and metabolites, these materials support a better understanding of the relationship between dietary intake and overall nutritional status.

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