Biochemistry Department (0308), 304 Engel Hall, Virginia Tech, Blacksburg, VA 24061, United States.
Food Chem. 2024 May 1;439:138091. doi: 10.1016/j.foodchem.2023.138091. Epub 2023 Dec 16.
A robust method for quantitation of total vitamin D and D in mushrooms by high performance liquid chromatography with UV detection (HPLC-UV) was developed to analyze mushrooms exposed to UV light. A two-step solid phase extraction (SPE) (silica, carbon black) removed chromatographic interferences typically resolved only with mass spectrometric detection (LC-MS) and allowed quantitation of all vitamin D and pre-D analytes. The vitamin and pre-vitamin forms of D, D and D (internal standard), as well as other photoisomers and sterols were resolved. Results for six types of UV-exposed mushrooms were comparable to LC-MS. Screening of ten additional types of UV-exposed mushrooms without the IS confirmed lack of interference with the IS. The limit of quantification (µg/100 g fresh weight) was 0.4 for vitamin D and 0.9 for pre-vitamin D. Mushrooms do not have to be dried, and separatory funnels and large solvent volumes were also eliminated from sample preparation.
建立了一种采用高效液相色谱-紫外检测（HPLC-UV）检测蘑菇中总维生素 D 和 D 的稳健方法，用于分析暴露于紫外光下的蘑菇。两步固相萃取（SPE）（硅胶、炭黑）去除了通常仅通过质谱检测（LC-MS）才能解决的色谱干扰，并允许定量分析所有维生素 D 和前 D 分析物。维生素 D 和前维生素 D（内标）以及其他光异构体和甾醇的形式都得到了分离。六种经 UV 处理的蘑菇的结果与 LC-MS 相当。对十种未经 IS 筛选的额外类型的经 UV 处理的蘑菇进行测试，确认 IS 没有干扰。定量限（µg/100 g 鲜重）为维生素 D 为 0.4，前维生素 D 为 0.9。蘑菇无需干燥，且样品制备中也省去了分液漏斗和大量溶剂。
