title: Exogenous versus endogenous recovery of 25-hydroxyvitamins D2 and D3 in human samples using high-performance liquid chromatography and the DiaSorin LIAISON Total-D Assay.
abstract: Demand for circulating 25-hydroxyvitamin D [25(OH)D] measurements has exploded due to its relationship with many serious health problems. The present study was designed to investigate the validity of samples "spiked" with 25-hydroxyvitamin D2 [25(OH)D2] or 25-hydroxyvitamin D3 [25(OH)D3] to determine their analytical recovery by the DiaSorin LIAISON 25 OH Vitamin D Total Assay (DiaSorin Assay) and high-performance liquid chromatography (HPLC). 25(OH)D was measured in nine volunteers taking large daily doses of vitamin D2 for 2 weeks. Samples were obtained pre-supplementation and 1 week following vitamin D2. Pre-supplementation samples were used for exogenous recovery studies by adding 25(OH)D2 or 25(OH)D3. Endogenous 25(OH)D [25(OH)D2 plus 25(OH)D3] concentrations reported by the DiaSorin Assay or detected by HPLC were in excellent agreement. However, exogenously added 25(OH)D2 and 25(OH)D3 were under-recovered by the DiaSorin Assay. NIST vitamin D standards containing serum from another species (horse) or exogenous 25(OH)D2 were similarly affected when using the DiaSorin Assay. Exogenous 25(OH)D2, 25(OH)D3 or serum from other species added to human samples is inappropriate in determining the analytical recovery of vitamin D compounds when using the DiaSorin Assay. Only endogenous 25(OH)D2 and/or 25(OH)D3 contained in human blood samples should be utilized for this purpose.
authors.affiliation: Heartland Assays Inc., 2325 North Loop Dr., Suite 6300, Ames, IA 50010, United States. Ron.Horst@heartlandassays.com
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Horst
mesh_terms: D015652:25-Hydroxyvitamin D 2 / Q000097:blood* / Q000494:pharmacology; D000818:Animals; D001681:Biological Assay; D002112:Calcifediol / Q000097:blood* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards*; D019587:Dietary Supplements; D004305:Dose-Response Relationship, Drug; D006736:Horses; D006801:Humans; D015203:Reproducibility of Results; D013048:Specimen Handling; D013696:Temperature
chemical_list: D015652:25-Hydroxyvitamin D 2; D002112:Calcifediol