Quantitation of total vitamin D2 and D4 in UV-exposed mushrooms using HPLC with UV detection after novel two-step solid phase extraction

• Published in: Food chemistry 2024-05, Vol.439, p.138091-138091, Article 138091
• Main Authors: McGinty, Ryan C., Phillips, Katherine M.
• Format: Article
• Language: English
• ISSN: 0308-8146; 1873-7072
• DOI: 10.1016/j.foodchem.2023.138091

A robust method for quantitation of total vitamin D2 and D4 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 D2, D4 and D3 (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.