Comprehensive Determination of Amino Acids for Diagnosis of Inborn Errors of Metabolism

Stephen M. Roper, Annette L. Weindel, Dennis J. Dietzen

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

Analysis of clinically relevant amino acids using ion exchange chromatography coupled to photometric/fluorescent detection has been an indispensable component in the detection of inborn errors of metabolism for six decades. Detection of amino acids using mass spectrometry offers advantages in speed and analytic specificity. Employing methanol extraction and controlled butylation, C8 reversed-phase chromatography, and MS/MS detection, 32 amino acids are quantified in 20 min with clinically appropriate imprecision in plasma, urine, and cerebrospinal fluid (CSF). Quantitation is linear to 2500 μM, and limits of detection are at least 1.0 μM. Important isobaric amino acids are distinguished by chromatography or by unique patterns of fragmentation following collision-induced dissociation (CID). The technique employs commercially available reagents and may be expanded and customized for specific clinical or research settings.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages55-64
Number of pages10
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2546
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Amino acid
  • Butylation
  • Isobar
  • Liquid chromatography
  • Mass spectrometry
  • Multiple reaction monitoring
  • Newborn screening

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