title: Degradation studies of cholecalciferol (vitamin D3) using HPLC-DAD, UHPLC-MS/MS and chemical derivatization.
abstract: In any food fortification program, the stability of added micronutrients is an important factor. Cholecalciferol or vitamin D3 is known to isomerise under various conditions, thereby making its analysis challenging. In the current study, the effects of different parameters, such as temperature, iodine, acidic conditions, and oxidation, on the isomerisation of vitamin D3 were studied using HPLC-DAD and UHPLC-MS/MS. Vitamin D3 thermally and reversibly transforms to pre-vitamin D3 type isomers. In the presence of iodine, cis/trans isomerisation of both cholecalciferol and pre-vitamin D3 takes place to form trans-vitamin D3 and tachysterol, respectively. Another isomer, isotachysterol, was formed under acidic conditions. The different rates of reaction of these products with a dienophile through the Diels-Alder reaction confirmed the formation of vitamin D3 isomerisation products. The derivatization enhanced the ionisation efficiency of vitamin D3 and its isomers in UHPLC-MS/MS and improved the separation and fragmentation enabling sensitive detection.
authors.affiliation: School of Chemical Sciences, Food Science Programme, University of Auckland, New Zealand.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Mahmoodani
authors.affiliation: School of Chemical Sciences, Food Science Programme, University of Auckland, New Zealand. Electronic address: conradperera@gmail.com.
authors.forename: Conrad O
authors.initials: CO
authors.lastname: Perera
authors.affiliation: School of Chemical Sciences, Food Science Programme, University of Auckland, New Zealand.
authors.forename: Bruno
authors.initials: B
authors.lastname: Fedrizzi
authors.affiliation: Fonterra Cooperative Group Ltd, Palmerston North, New Zealand.
authors.forename: Grant
authors.initials: G
authors.lastname: Abernethy
authors.affiliation: Fonterra Cooperative Group Ltd, Palmerston North, New Zealand.
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
mesh_terms: D002762:Cholecalciferol / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D004875:Ergosterol / Q000031:analogs & derivatives / Q000032:analysis; D005527:Food, Fortified / Q000032:analysis*; D007536:Isomerism; D053719:Tandem Mass Spectrometry
chemical_list: D002762:Cholecalciferol; C002960:isotachysterol; D004875:Ergosterol
keywords: 4-Phenyl-1,2,4-triazoline-3,5-dione (PubChem CID: 77913); 5,6-trans-cholecalciferol (PubChem CID: 12303099); Cholecalciferol; Cholecalciferol (PubChem CID: 5280795); Degradation; Isomerization; Isotachysterol3 (PubChem CID: 5283714); Pre-cholecalciferol (PubChem CID: 5281058); Tachysterol3 (PubChem CID: 5283713); UHPLC-MS/MS; Vitamin D3