Latest Publication—4th January 2019

Significance of previtamin D chromatographic resolution in the accurate determination of vitamin D3 by HPLC‑UV

David C. Woollard, Harvey E. Indyk, Brendon D. Gill

Conventional methods using HPLC with UV detection have used vitamin D2 as an internal standard with the expectation that this fully compensates for the heat-dependent equilibrium of vitamin D3 with its previtamin. Previtamin D has a different spectral absorptivity from vitamin D and may be present in different proportions in samples and standards. Therefore, vitamin D2 and vitamin D3 and their previtamin forms must be chromatographically resolved to achieve accurate quantitation of total vitamin D. This study identified four chromatographic columns (ACE C18, ACE C18 AR, Vydac 201 TP C18 and Polaris C18-Ether) with adequate selectivity that should be applied for food testing and further confirmed that both parent vitamins isomerise at the same rate under thermal conditions.

Woollard, D.C., Indyk, H.E., Gill, B.D. (2019) J. Food Comp. Anal. 79, 1‑4
doi: 10.1016/j.jfca.2019.02.010

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