title: Quantification of 1,25-Dihydroxyvitamin D2 and D3 in Serum Using Liquid Chromatography-Tandem Mass Spectrometry.
abstract: 1,25-Dihydroxyvitamin D is the active form of vitamin D and plays a critical role in the maintenance of calcium and phosphorous metabolism of the human body. Measurement of 1,25-dihydroxyvitamin D in serum can aid in clinical diagnosis and/or management of renal disease, sarcoidosis, and rare inherited diseases. We present here an effective and accurate method for measuring 1,25-dihydroxyvitamin D3 and 1,25-dihydroxyvitamin D2 by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) after immunoaffinity extraction. The MS/MS is operated in multiple reaction mode with positive electrospray. Quantification is based on peak area ratios of the analytes to respective deuterated internal standards. This method offered a linear range from 4.0 to 160.0 pg/mL with analytical recovery of 89.9-115.5 % for both 1,25-dihydroxyvitamin D3 and 1,25-dihydroxyvitamin D2.
authors.affiliation: Department of Laboratory Medicine, Cleveland Clinic, Cleveland, OH, USA.
authors.forename: Jonathon
authors.initials: J
authors.lastname: Mahlow
authors.affiliation: Department of Laboratory Medicine, Cleveland Clinic, Cleveland, OH, USA.|Department of Chemistry, Cleveland State University, Cleveland, OH, USA.
authors.forename: Dustin R
authors.initials: DR
authors.lastname: Bunch
authors.affiliation: Department of Laboratory Medicine, Cleveland Clinic, Cleveland, OH, USA. wangs2@ccf.org.
authors.forename: Sihe
authors.initials: S
authors.lastname: Wang
mesh_terms: D000085:Acetates / Q000737:chemistry; D001774:Blood Chemical Analysis / Q000379:methods*; D002117:Calcitriol / Q000097:blood* / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D004872:Ergocalciferols / Q000097:blood* / Q000737:chemistry; D006801:Humans; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000085:Acetates; D004872:Ergocalciferols; C003564:1,25-dihydroxyergocalciferol; D002117:Calcitriol; C488804:lithium acetate
keywords: 1,25-Dihydroxyvitamin D; Liquid chromatography; Mass spectrometry; Quantification; Serum