Title

Quantification of 25-Hydroxyvitamin D2 and D3 Using Liquid Chromatography-Tandem Mass Spectrometry.

Document Type

Article

Publication Date

2022

Identifier

DOI: 10.1007/978-1-0716-2565-1_48

Abstract

Vitamin D plays an important role not only in bone health but also in many other body functions. Vitamin D deficiency is very common in the general population. Measurement of blood 25-hydroxyvitamin D is a common practice to evaluate vitamin D deficiency. Immunoassays and liquid chromatography tandem mass spectrometry (LC-MS/MS) are the most commonly used methods for the measurement of 25-hydroxyvitamin D. Immunoassays suffer from specificity issues and do not distinguish between 25-hydroxyvitamin D2 and D3. Therefore, LC-MS/MS is a preferred method for quantification of 25-hydroxyvitamin. We describe an LC-MS/MS method, which involves protein precipitation and analysis of the extract using atmospheric pressure chemical ionization and multiple reaction monitoring. 25-hydroxyvitamin D3-d6 is used as an internal standard. The method is linear from 1-100 ng/mL for both 25-hydroxyvitamin D2 and D3 and has imprecision of <10%.

Journal Title

Methods in molecular biology (Clifton, N.J.)

Volume

2546

First Page

539

Last Page

544

MeSH Keywords

25-Hydroxyvitamin D 2; Calcifediol; Chromatography, Liquid; Humans; Tandem Mass Spectrometry; Vitamin D; Vitamin D Deficiency

Keywords

25-Hydroxyvitamin D2; 25-Hydroxyvitamin D3; Tandem Mass Spectrometry; Vitamin D

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