Amino Acid Analysis pp 1-11

Part of the Methods in Molecular Biology book series (MIMB, volume 828) | Cite as

Rapid LC–MS/MS Profiling of Protein Amino Acids and Metabolically Related Compounds for Large-Scale Assessment of Metabolic Phenotypes

Protocol

Abstract

Amino acids extracted from a biological matrix can be resolved and measured using a 6-min per sample method through high-performance liquid chromatography with a short C18 column and rapid gradient using the ion-pairing reagent perfluoroheptanoic acid. LC-tandem mass spectrometry with multiple reaction monitoring (MRM) transitions selective for each compound allows simultaneous quantification of the 20 proteinogenic amino acids and 5 metabolically related compounds. Distinct MRM transitions were also established for selective detection of the isomers leucine/isoleucine and threonine/homoserine.

Key words

High-performance liquid chromatography-tandem mass spectrometry Ion-pairing reagent Multiple reaction monitoring transition Underivatized amino acids Internal standard Amino acid quantification Amino acid analysis 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Biology and MicrobiologySouth Dakota State UniversityBrookingsUSA
  2. 2.Department of Biochemistry and Molecular Biology, and Department of ChemistryMichigan State UniversityEast LansingUSA
  3. 3.Departments of Biochemistry and Molecular Biology and Department of Plant BiologyMichigan State UniversityEast LansingUSA

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