Phillips, Katherine M.Horst, R. L.Koszewski, N. J.Simon, R. R.2017-01-162017-01-162012-08-031932-6203http://hdl.handle.net/10919/74350An unknown vitamin D compound was observed in the HPLC-UV chromatogram of edible mushrooms in the course of analyzing vitamin D<sub>2</sub> as part of a food composition study and confirmed by liquid chromatography-mass spectrometry to be vitamin D<sub>4</sub> (22-dihydroergocalciferol). Vitamin D<sub>4</sub> was quantified by HPLC with UV detection, with vitamin [<sub>3</sub>H] vitamin D<sub>3</sub> as an internal standard. White button, crimini, portabella, enoki, shiitake, maitake, oyster, morel, chanterelle, and UV-treated portabella mushrooms were analyzed, as four composites each of a total of 71 samples from U.S. retail suppliers and producers. Vitamin D<sub>4</sub> was present (>0.1 μg/100 g) in a total of 18 composites and in at least one composite of each mushroom type except white button. The level was highest in samples with known UV exposure: vitamin D enhanced portabella, and maitake mushrooms from one supplier (0.2–7.0 and 22.5–35.4 μg/100 g, respectively). Other mushrooms had detectable vitamin D<sub>4</sub> in some but not all samples. In one composite of oyster mushrooms the vitamin D<sub>4</sub> content was more than twice that of D<sub>2</sub> (6.29 vs. 2.59 μg/100 g). Vitamin D<sub>4</sub> exceeded 2 μg/100 g in the morel and chanterelle mushroom samples that contained D<sub>4</sub>, but was undetectable in two morel samples. The vitamin D<sub>4</sub> precursor 22,23- dihydroergosterol was found in all composites (4.49–16.5 mg/100 g). Vitamin D<sub>4</sub> should be expected to occur in mushrooms exposed to UV light, such as commercially produced vitamin D enhanced products, wild grown mushrooms or other mushrooms receiving incidental exposure. Because vitamin D<sub>4</sub> coeluted with D<sub>3</sub> in the routine HPLC analysis of vitamin D<sub>2</sub> and an alternate mobile phase was necessary for resolution, researchers analyzing vitamin D<sub>2</sub> in mushrooms and using D<sub>3</sub> as an internal standard should verify that the system will resolve vitamins D<sub>3</sub> and D<sub>4</sub>.10 pagesapplication/pdfenCreative Commons Attribution 4.0 Internationalagaricus-bisporusbutton mushroomsedible mushroomsultraviolet-irradiationcultivated mushroomsbiological-activitysterolsergosterolmammarywildVitamin D-4 in MushroomsArticle - RefereedPLOS ONEhttps://doi.org/10.1371/journal.pone.004070278