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Amino Acid Analysis by Hydrophilic Interaction Chromatography Coupled with Isotope Dilution Mass Spectrometry

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2030))

Abstract

Here, we describe an amino acid analysis that is based on the use of hydrophilic interaction liquid chromatography coupled with isotope dilution mass spectrometry for the accurate quantification of underivatized amino acids from hydrolyzed peptide/protein. Twelve underivatized amino acids were separated and detected during an 88-min runtime. The absolute limits of detection and limits of quantification (on column) of the four amino acids (isoleucine, phenylalanine, proline, and valine) were in the range of 6–80 and 20–200 fmol, respectively. As little as 25 pmol of peptide or protein hydrolysate is sufficient for determining absolute content.

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Correspondence to Megumi Kato .

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Kato, M., Takatsu, A. (2019). Amino Acid Analysis by Hydrophilic Interaction Chromatography Coupled with Isotope Dilution Mass Spectrometry. 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_9

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  • DOI: https://doi.org/10.1007/978-1-4939-9639-1_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9638-4

  • Online ISBN: 978-1-4939-9639-1

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