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.
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References
Wang S (2009) Epidemiology of vitamin D in health and disease. Nutr Res Rev 22:188–203
DeLuca HF (2008) Evolution of our understanding of vitamin D. Nutr Rev 66:S73–S87
El-Khoury JM, Reineks EZ, Wang S (2011) Progress of liquid chromatography-mass spectrometry in measurement of vitamin D metabolites and analogues. Clin Biochem 44:66–76
Burtis CA, Ashwood ER, Burns DE (2006) Tietz textbook of clinical chemistry and molecular diagnostics, 4th edn. W.B. Saunders, Philadelphia
Carter GD (2009) 25-Hydroxyvitamin D assays: the quest for accuracy. Clin Chem 55:1300–1302
Aronov PA, Hall LM, Dettmer K, Stephensen CB, Hammock BD (2008) Metabolic profiling of major vitamin D metabolites using Diels-Alder derivatization and ultra-performance liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 391:1917–1930
Duan X, Weinstock-Guttman B, Wang H, Bang E, Li J, Ramanathan M, Qu J (2010) Ultrasensitive quantification of serum vitamin D metabolites using selective solid-phase extraction coupled to microflow liquid chromatography and isotope-dilution mass spectrometry. Anal Chem 82:2488–2497
Kissmeyer AM, Sonne K (2001) Sensitive analysis of 1alpha,25-dihydroxyvitamin D3 in biological fluids by liquid chromatography-tandem mass spectrometry. J Chromatogr A 935:93–103
Casetta B, Jans I, Billen J, Vanderschueren D, Bouillon R (2010) Development of a method for the quantification of 1alpha,25(OH)2-vitamin D3 in serum by liquid chromatography tandem mass spectrometry without derivatization. Eur J Mass Spectrom (Chichester, Eng) 16:81–9
Yuan C, Kosewick J, He X, Kozak M, Wang S (2011) Sensitive measurement of serum 1alpha,25-dihydroxyvitamin D by liquid chromatography/tandem mass spectrometry after removing interference with immunoaffinity extraction. Rapid Commun Mass Spectrom 25:1241–1249
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Mahlow, J., Bunch, D.R., Wang, S. (2016). Quantification of 1,25-Dihydroxyvitamin D2 and D3 in Serum Using Liquid Chromatography-Tandem Mass Spectrometry. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 1378. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3182-8_31
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DOI: https://doi.org/10.1007/978-1-4939-3182-8_31
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3181-1
Online ISBN: 978-1-4939-3182-8
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