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Quantification of 1,25-Dihydroxyvitamin D2 and D3 in Serum Using Liquid Chromatography-Tandem Mass Spectrometry

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Clinical Applications of Mass Spectrometry in Biomolecular Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1378))

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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Correspondence to Sihe Wang Ph.D. .

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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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