Mass Spectrometry Assays of Vitamin D Metabolites

Martin Kaufmann, Lusia Sepiashvili, Ravinder Jit Singh

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations


Liquid chromatography tandem mass spectrometry (LC-MS/MS) has emerged as the gold standard technique with the capability for reliable, accurate, and high-throughput quantification of circulating vitamin D metabolites. The complexity of vitamin D metabolism has driven the continuous development of the basic steps involved in LC-MS/MS methodology including sample extraction, derivatization, separation, and mass spectrometry, used to interrogate the role of vitamin D nutrition and metabolism in human health and disease states. From the perspective of both the high-throughput clinical chemistry laboratory and basic research applications, this chapter describes how LC-MS/MS can be optimized to provide sensitive and specific measurements of 25-hydroxyvitamin D, along with other low-abundance metabolites including 24-25-dihydroxyvitamin D and 1,25-dihydroxyvitamin D, simultaneously. Furthermore, we describe the advantages of using LC-MS/MS in studying mouse models of vitamin D metabolism and how this work has contributed to an improved understanding of the pathophysiology of certain vitamin D-related disease states.

Original languageEnglish (US)
Title of host publicationBiochemistry, Physiology and Diagnostics
PublisherElsevier Inc.
Number of pages15
ISBN (Electronic)9780128099667
ISBN (Print)9780128099650
StatePublished - Dec 18 2017


  • Chemical derivatization
  • Hypercalcemia
  • Knockout mouse
  • Liquid chromatography tandem mass spectrometry
  • Vitamin D metabolite

ASJC Scopus subject areas

  • General Agricultural and Biological Sciences
  • General Biochemistry, Genetics and Molecular Biology
  • General Medicine


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