Abstract
One of the aims of untargeted metabolomics is searching for selective biomarkers of different pathophysiological conditions. Modified amino acids originated from the posttranslational modification of proteins play a key role as potential biomarkers; however, they are very often still classified as unknown after metabolite annotation. We have developed an analytical workflow for the targeted screening of these compounds using CE-ESI-MS. The workflow is based on the in-source fragmentation of molecules that produces diagnostic ions that we have collected in an open-source library. In this chapter, we describe in detail the strategy for the targeted screening of modified amino acids (MAAs), using as an example l-proline and its modified derivatives. We illustrate the strategy with two case studies in human plasma.
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
Mamani-Huanca M, Gradillas A, Gil de la Fuente A, López-Gonzálvez A, Barbas C (2020) Unveiling the fragmentation mechanisms of modified amino acids as the key for their targeted identification. Anal Chem 92(7):4848–4857. https://doi.org/10.1021/acs.analchem.9b04313
Krane SM (2008) The importance of proline residues in the structure, stability, and susceptibility to proteolytic degradation of collagens. Amino Acids 35(4):703–710. https://doi.org/10.1007/s00726-008-0073-2
Hulmes DJS (2008) Collagen diversity, synthesis and assembly. In: Fratzl P (ed) Collagen. Springer, Boston, MA, pp 15–47. https://doi.org/10.1007/978-0-387-73906-9_2
Rebholz CM, Lichtenstein AH, Zheng Z, Appel LJ, Coresh J (2018) Serum untargeted metabolomic profile of the Dietary Approaches to Stop Hypertension (DASH) dietary pattern. Am J Clin Nutr 108(2):243–255. https://doi.org/10.1093/ajcn/nqy099
Hu BH, Martin M (1999) Capillary electrophoresis of peptides and proteins. In: Millner P (ed) High-resolution chromatography: a practical approach school of biochemistry and molecular biology. University of Leeds, Leeds, pp 77–118. ISBN 10: 0199636486
Domingo-Almenara X, Montenegro-Burke JR, Guijas C, Majumder ELW, Benton HP, Siuzdak G (2019) Autonomous METLIN-guided in-source fragment annotation for untargeted metabolomics. Anal Chem 91(5):3246–3253. https://doi.org/10.1021/acs.analchem.8b03126
Mamani-Huanca M, Gil de la Fuente A, Otero A, Gradillas A, Godzien J, Barbas C, López-Gonzálvez A (2021) Enhancing confidence of metabolite annotation in capillary electrophoresis-mass spectrometry untargeted metabolomics with relative migration time and in-source fragmentation. J Chromatogr A 1635:461758. https://doi.org/10.1016/j.chroma.2020.461758
De Vijlder T, Valkenborg D, Lemière F, Romijn EP, Laukens K, Cuyckens F (2018) A tutorial in small molecule identification via electrospray ionization-mass spectrometry: the practical art of structural elucidation. Mass Spectrom Rev 37(5):607–629. https://doi.org/10.1002/mas.21551
Parcher JF, Wang M, Chittiboyina AG, Khan IA (2018) In-source collision-induced dissociation (IS-CID): applications, issues, and structure elucidation with single-stage mass analyzers. Drug Test Anal 10(1):28–36. https://doi.org/10.1002/dta.2249
Zhang P, Chan W, Ang IL, Wei R, Lam MMT, Lei KMK et al (2019) Revisiting fragmentation reactions of protonated α-amino acids by high-resolution electrospray ionization tandem mass spectrometry with collision-induced dissociation. Sci Rep 9(1):6453. https://doi.org/10.1038/s41598-019-42777-8
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Mamani-Huanca, M., Gradillas, A., López-Gonzálvez, Á., Barbas, C. (2022). In-Source Fragmentation for the Identification of Compounds by CE-ESI-TOF in Human Plasma. l-Proline as Case Study. In: Neusüß, C., Jooß, K. (eds) Capillary Electrophoresis-Mass Spectrometry . Methods in Molecular Biology, vol 2531. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2493-7_12
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2493-7_12
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2492-0
Online ISBN: 978-1-0716-2493-7
eBook Packages: Springer Protocols