Abstract
Liquid-chromatography tandem mass spectrometry (LC-MS/MS) has been shown to be an effective approach in the clinical analysis of 25-OH-vitamin D in patient serum. Test volumes vary among laboratories and different levels of throughput are required for different settings. LC-MS/MS assays with multiple LC channels can be beneficial for labs with the demand of large sample volume (e.g., 300 or more samples) to control costs and fulfill a reasonable turnaround time. We hereby present an assay that employs 4 LC channels (4-plex), which are coupled to the TSQ Endura triple-quadrupole (QqQ) MS instrument, for a high-throughput solution. Briefly, the pre- and postelution segments of the LC gradient are diverted to waste via solenoid valve controls, reserving the data acquisition for only the elution segment per injection per channel at a time. The multiplexing affords a manifold increase in throughput and the optimization of the duty cycle, without compromise in assay performance and precision.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Park H, Brannon PM, West AA, Yan J, Jiang X, Perry CA, Malysheva OV, Mehta S, Caudill MA (2016) Vitamin D metabolism varies among women in different reproductive states consuming the same intakes of vitamin D and related nutrients. J Nutr 146(8):1537–1545. https://doi.org/10.3945/jn.116.229971
Hollis BW (2005) Circulating 25-hydroxyvitamin D levels indicative of vitamin D sufficiency: implications for establishing a new effective dietary intake recommendation for vitamin D. J Nutr 135(2):317–322. https://doi.org/10.1093/jn/135.2.317
Farrell CJ, Martin S, McWhinney B, Straub I, Williams P, Herrmann M (2012) State-of-the-art vitamin D assays: a comparison of automated immunoassays with liquid chromatography-tandem mass spectrometry methods. Clin Chem 58(3):531–542. https://doi.org/10.1373/clinchem.2011.172155
Bedner M, Lippa KA, Tai SS (2013) An assessment of 25-hydroxyvitamin D measurements in comparability studies conducted by the vitamin D metabolites quality assurance program. Clin Chim Acta 426:6–11. https://doi.org/10.1016/j.cca.2013.08.012
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(5):1917–1930. https://doi.org/10.1007/s00216-008-2095-8
Bedner M, Phinney KW (2012) Development and comparison of three liquid chromatography-atmospheric pressure chemical ionization/mass spectrometry methods for determining vitamin D metabolites in human serum. J Chromatogr A 1240:132–139. https://doi.org/10.1016/j.chroma.2012.03.091
Shah I, James R, Barker J, Petroczi A, Naughton DP (2011) Misleading measures in vitamin D analysis: a novel LC-MS/MS assay to account for epimers and isobars. Nutr J 10:46. https://doi.org/10.1186/1475-2891-10-46
Gervasoni J, Cocci A, Zuppi C, Persichilli S (2013) Total 25-hydroxyvitamin D determination by an entry level triple quadrupole instrument: comparison between two commercial kits. Biomed Res Int 2013:270426. https://doi.org/10.1155/2013/270426
Stone J (2016) High-throughput serum 25-Hydroxy vitamin D testing with automated sample preparation. Methods Mol Biol 1378:301–320. https://doi.org/10.1007/978-1-4939-3182-8_32
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Gqamana, P.P., Zhang, Y.V. (2022). High-Throughput Analysis of 25-OH-Vitamin D2 and D3 Using Multichannel Liquid Chromatography-Tandem Mass Spectrometry. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 2546. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2565-1_47
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2565-1_47
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2564-4
Online ISBN: 978-1-0716-2565-1
eBook Packages: Springer Protocols