Development Validation and Application of HPLC Method for Simultaneous Analysis of Vitamin D2 D3 and Precursors in Natural Sources

Abstract

Vitamin D is an essential compound necessary for overall health and newlinephysiological function. It promotes calcium and phosphorus metabolism by improving their newlineintestinal absorption and maintaining normal blood levels. Besides its function in bone newlinehealth, vitamin D regulates cell proliferation and differentiation and alters immune responses newlineby suppressing pro-inflammatory cytokines and enhancing anti-inflammatory ones. newlineVitamin D exists in two principal forms: Vitamin D2 (ergocalciferol) and newlineVitamin D3 (cholecalciferol). Vitamin D2 is obtained from dietary sources, such as fortified newlinefoods and supplements, while vitamin D3 is produced in the skin through UVB radiation newlineexposure. Ergosterol and 7-dehydrocholesterol serve as precursors for these variants of newlinevitamin D. Ergosterol is abundant in mushrooms due to their fungal origin but is present in newlinecomparatively low concentrations in vegetable oils. In contrast, 7-dehydrocholesterol is newlinepredominantly present in animal tissues, especially the skin, and is found in negligible newlineamounts in specific vegetable oils. These precursors collectively offer potential pathways to newlineenhance vitamin D availability from diverse sources. newlineAssessing vitamin D in various sources is challenging because of the low newlineconcentrations and the presence of many active vitamin D molecules, particularly in plant newlinematerials with intricate constituents. Historically, bioassay methods using animal-based newlineprotocols were utilized to evaluate vitamin D levels. These methods are arduous, requiring newlineat least five weeks, and lack precision as they evaluate overall Vitamin D activity without newlinedistinguishing specific metabolites newline