Determination of vitamin D3 and 25-hydroxyvitamin D3 in foodstuffs by HPLC UV-DAD and LC-MS/MS

In order to generate new data for vitamin D content for the Canadian Nutrient File, a method for the quantification of vitamin D-3 and 25(OH)D-3 in foodstuffs has been modified and improved. Vitamin D-3 was quantified using reverse phase liquid chromatography (LC) with UV-diode array detector (UV-DAD), while 25(OH)D-3 was measured by triple quadrupole mass spectrometry (APCI MS/MS). Quantification was by internal standards (IS) using vitamin D-2 and 25(OH)D-2. A Certified Reference Material (CRM-421 containing vitamin D-3) and a control sample (internally generated reference material of ground pork containing both vitamin D-3 and 25(OH)D-3) were used as validation and quality control tools. Limit of detection for both compounds was 0.04 mu g/100 g. Accuracy for vitamin D in the CRM-421 was 99% (0.142 mg/kg for a target of 0.143, n = 10). Recovery of vitamin D-3 in ground pork was 97% (88% absolute recovery). For 25(OH)D-3, a recovery of 94% (73% absolute recovery) was obtained. Using this method, data for vitamin D-3 and 25(OH)D-3 content in a variety of foods (pork, beef, eggs, poultry, fish, and dinners) have been generated. Crown Copyright (C) 2010 Published by Elsevier Inc. All rights reserved.