Chapter title |
Quantification of 1,25-Dihydroxyvitamin D2 and D3 in Serum Using Liquid Chromatography-Tandem Mass Spectrometry
|
---|---|
Chapter number | 31 |
Book title |
Clinical Applications of Mass Spectrometry in Biomolecular Analysis
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3182-8_31 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3181-1, 978-1-4939-3182-8
|
Authors |
Jonathon Mahlow, Dustin R. Bunch, Sihe Wang |
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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