This European Standard specifies a method for the determination of vitamin D3 (cholecalciferol) or vitamin D2 (ergocalciferol) in foodstuffs by high performance liquid chromatography (HPLC). Vitamin D3 is primary in foodstuffs of animal origin, while vitamin D2 is primary in wild mushrooms. Both vitamin D3 and vitamin D2 can be present in fortified foodstuffs. This European Standard is not applicable for samples with a content of vitamin D3 and vitamin D2. Apart from the vitamin D activity from the parent forms, vitamin D3 and vitamin D2, the corresponding metabolites 25-hydroxy vitamin D and 1,25-dihydroxy vitamin D also contribute to the vitamin D activity. This European Standard does only include measurement of vitamin D3 or vitamin D2. This European Standard provides the base for the analytical methods. It is intended to serve as a frame in which the analyst can define his own analytical work in accordance to the standard procedure. This method has been validated in inter-laboratory tests on fortified and non-fortified samples such as margarine, milk, milk powder, liquid infant formula, infant formula, cooking oil, and fish oil at levels from 0,4 µg/100 g to 14 µg/100 g. Further information on the validation data is given in Annex D.
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