Quantification of 25-Hydroxyvitamin D2 and D3 Using Liquid Chromatography-Tandem Mass Spectrometry.
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%.
Methods in molecular biology (Clifton, N.J.)
25-Hydroxyvitamin D 2; Calcifediol; Chromatography, Liquid; Humans; Tandem Mass Spectrometry; Vitamin D; Vitamin D Deficiency
25-Hydroxyvitamin D2; 25-Hydroxyvitamin D3; Tandem Mass Spectrometry; Vitamin D
Munar A, Clinton Frazee C 3rd, Garg U. Quantification of 25-Hydroxyvitamin D2 and D3 Using Liquid Chromatography-Tandem Mass Spectrometry. Methods Mol Biol. 2022;2546:539-544. doi:10.1007/978-1-0716-2565-1_48