Vitamin D(2) (ergocalciferol) and sterols were analyzed in mushrooms sampled nationwide in the United States to update the USDA Nutrient Database for Standard Reference. Vitamin D(2) was assayed using HPLC with [(3)H]-vitamin D(3) internal standard and sterols by GC-FID mass spectrometric (MS) confirmation. Vitamin D(2) was low (0.1-0.3 μg/100 g) in Agaricus bisporus (white button, crimini, portabella) and enoki, moderate in shiitake and oyster (0.4-0.7 μg/100 g), and high in morel, chanterelle, maitake (5.2-28.1 μg/100 g) and UV-treated portabella (3.4-20.9 μg/100 g), with significant variability among composites for some types. Ergosterol (mg/100 g) was highest in maitake and shiitake (79.2, 84.9) and lowest in morel and enoki (26.3, 35.5); the range was <10 mg/100 g among white button composites but 12-50 mg/100 g among samples of other types. All mushrooms contained ergosta-5,7-dienol (22,23-dihydroergosterol) (3.53-18.0 mg/100 g) and (except morel) ergosta-7-enol. Only morel contained brassicasterol (28.6 mg/100 g) and campesterol (1.23-4.54 mg/100 g) and no ergosta-7,22-dienol. MS was critical in distinguishing campesterol from ergosta-7,22-dienol.
This study compared the compositional changes in mushrooms exposed to sunlight with those occurring after commercial ultraviolet (UV) light processing. Button mushrooms (75 kg) were processed in the presence or absence of UVB light; a third group was exposed to direct sunlight. Mushroom composition was evaluated using chemical analyses. Vitamin D concentrations were 5, 410, and 374 μg/100 g (dw) in control, UVB, and sunlight groups, respectively. On a dry weight basis, no significant changes in vitamin C, folate, vitamins B(6), vitamin B(5), riboflavin, niacin, amino acids, fatty acids, ergosterol, or agaritine were observed following UVB processing. Sunlight exposure resulted in a 26% loss of riboflavin, evidence of folate oxidation, and unexplained increases in ergosterol (9.5%). It was concluded that compositional effects of UVB light are limited to changes in vitamin D and show no detrimental changes relative to natural sunlight exposure and, therefore, provide important information relevant to the suitability and safety of UVB light technology for vitamin D enhanced mushrooms.
Bacillus subtilis CU1 is a recently described probiotic strain with beneficial effects on immune health in elderly subjects. The following work describes a series of studies supporting the safety of the strain for use as an ingredient in food and supplement preparations. Using a combination of 16S rDNA and gyrB nucleotide analyses, the species was identified as a member of the Bacillus subtilis complex (B. subtilis subsp. spizizenii). Further characterization of the organism at the strain level was achieved using random amplified polymorphic DNA polymerase chain reaction (RAPD PCR) and pulsed field gel electrophoresis (PFGE) analyses. B. subtilis CU1 did not demonstrate antibiotic resistance greater than existing regulatory cutoffs against clinically important antibiotics, did not induce hemolysis or produce surfactant factors, and was absent of toxigenic activity in vitro. Use of B. subtilis CU1 as a probiotic has recently been evaluated in a 16-week randomized, double-blind, placebo-controlled, parallel-arm study, in which 2 × 10 spores per day of B. subtilis CU1 were administered for a total 40 days to healthy elderly subjects (4 consumption periods of 10 days separated by 18-day washouts). This work describes safety related endpoints not previously reported. B. subtilis CU1 was safe and well-tolerated in the clinical subjects without undesirable physiological effects on markers of liver and kidney function, complete blood counts, hemodynamic parameters, and vital signs.
An unknown vitamin D compound was observed in the HPLC-UV chromatogram of edible mushrooms in the course of analyzing vitamin D2 as part of a food composition study and confirmed by liquid chromatography-mass spectrometry to be vitamin D4 (22-dihydroergocalciferol). Vitamin D4 was quantified by HPLC with UV detection, with vitamin [3H] itamin D3 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 D4 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 D4 in some but not all samples. In one composite of oyster mushrooms the vitamin D4 content was more than twice that of D2 (6.29 vs. 2.59 µg/100 g). Vitamin D4 exceeded 2 µg/100 g in the morel and chanterelle mushroom samples that contained D4, but was undetectable in two morel samples. The vitamin D4 precursor 22,23-dihydroergosterol was found in all composites (4.49–16.5 mg/100 g). Vitamin D4 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 D4 coeluted with D3 in the routine HPLC analysis of vitamin D2 and an alternate mobile phase was necessary for resolution, researchers analyzing vitamin D2 in mushrooms and using D3 as an internal standard should verify that the system will resolve vitamins D3 and D4.
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