DIN EN 12823-1:2014-08

This European Standard specifies a method for the determination of vitamin A in foodstuffs by high performance liquid chromatography (HPLC). This determination has been validated in an interlaboratory study with samples of margarine and milk powder with all-E-retinol levels ranging from 653 µg/100 g to 729 µg/100 g and with 13-Z-retinol levels ranging from 30 µg/100 g to 39 µg/100 g. The determination of vitamin A content is carried out by the measurement of all-E-retinol, 13-Z-retinol and β-carotene. This part covers the measurement of all-E-retinol and 13-Z-retinol. The extract obtained in this method, after the saponification can be used for the determination of β-carotene, as described in EN 12823-2:2000 "Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 2: Measurements of β-carotene". In this case, the saponification temperature should preferably not exceed 80 °C in order to prevent isomerization and oxidation of β-carotene. In this standard, retinol is released by saponification by using methanolic or ethanolic potassium hydroxide solution; then retinol is extracted by an appropriate solvent and determined by high performance liquid chromatography (HPLC) with either fluorescence (F) or ultraviolet (UV) detection. The substances are identified on the basis of their retention times and determined by the external standard procedure using peak areas or heights. This document has been prepared by Technical Committee CEN/TC 275 "Food analysis - Horizontal methods", the secretariat of which is held by DIN (Germany). The responsible German body is Working Committee NA 057-01-13 AA "Vitamine und Carotinoide" ("Vitamins and carotinoids") at the Food and Agricultural Products Standards Committee (NAL) at DIN.