Abstract
The role of amino acid analysis in bioanalysis has changed from a qualitative to a quantitative technique. With the discovery of both electrospray ionization and matrix-assisted laser desorption ionization in the early 1990s, the use of amino acid analysis for qualitative analysis of proteins and peptides has been replaced by mass spectrometry. Accurate measurement of the relative molecular masses of proteins and peptides, peptide mapping, and sequencing by tandem mass spectrometry provide significantly better qualitative information than can be achieved from amino acid analysis. At NIST, amino acid analysis is used to assign concentration values to protein and peptide standard reference materials (SRMs) which, subsequently, will be used in the calibration of a wide variety of protein and peptide assays, such as those used in clinical diagnostics. It is critical that the amino acid analysis method used at NIST for assigning concentration values in SRM deliver the highest accuracy and precision possible. Therefore, we have developed an amino acid analysis method that uses isotope dilution LC-MS/MS—the analytical technique routinely used at NIST to certify analyte concentrations in SRMs for a wide variety of analytes. We present here our most recent method for the quantification of amino acids using isotope dilution LC-MS/MS.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Brun V et al (2009) Isotope dilution strategies for absolute quantitative proteomics. J Proteome 72:740–749
NIST Policy on Metrological Traceability. http://www.nist.gov/traceability/index.cfm. Accessed 28 Aug 2018
Burkitt WI et al (2008) Toward Système International d’Unité-traceable protein quantification: from amino acids to proteins. Anal Biochem 376:242–251
Sargent M, Harte R, Harrington C (2002) Guidelines for achieving high accuracy in isotope dilution mass spectrometry (IDMS). The Royal Society for Chemistry, London
Henrion A (1994) Reduction of systematic errors in quantitative analysis by isotope dilution mass spectrometry (IDMS): an iterative method. Fresenius J Anal Chem 350:657–658
Mackay LG et al (2003) High accuracy analysis by isotope dilution mass spectrometry using an iterative exact matching technique. Accred Qual Assur 8:191–194
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Bunk, D.M., Lowenthal, M.S. (2019). Isotope Dilution Liquid Chromatography-Tandem Mass Spectrometry for Quantitative Amino Acid Analysis. In: Alterman, M. (eds) Amino Acid Analysis. Methods in Molecular Biology, vol 2030. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9639-1_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9639-1_12
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9638-4
Online ISBN: 978-1-4939-9639-1
eBook Packages: Springer Protocols