Amino Acid Analysis pp 13-28

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

Combination of an AccQ·Tag-Ultra Performance Liquid Chromatographic Method with Tandem Mass Spectrometry for the Analysis of Amino Acids

  • Carolina Salazar
  • Jenny M. Armenta
  • Diego F. Cortés
  • Vladimir Shulaev
Protocol

Abstract

Amino acid analysis is a powerful tool in life sciences. Current analytical methods used for the detection and quantitation of low abundance amino acids in complex samples face intrinsic challenges such as insufficient sensitivity, selectivity, and throughput. This chapter describes a protocol that makes use of AccQ∙Tag chemical derivatization combined with the exceptional chromatographic resolution of ultra performance liquid chromatography (UPLC), and the sensitivity and selectivity of tandem mass spectrometry (MS/MS). The method has been fully implemented and validated using different tandem quadrupole detectors, and thoroughly tested for a variety of samples such as Plasmodium falciparum, human red blood cells, and Arabidopsis thaliana extracts. Compared to currently available methods for amino acid analysis, the AccQ∙Tag UPLC-MS/MS method presented here provides enhanced sensitivity and reproducibility, and offers excellent performance within a short analysis time and a broad dynamic range of analyte concentration. The focus of this chapter is the application of this improved protocol for the compositional amino acid analysis in A. thaliana leaf extracts using the Xevo TQ for mass spectrometric detection.

Key words

6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate AccQ∙Tag Amino acid analysis UPLC-ESI-MS/MS 

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Carolina Salazar
    • 1
  • Jenny M. Armenta
    • 2
  • Diego F. Cortés
    • 3
  • Vladimir Shulaev
    • 1
  1. 1.Department of Biological SciencesUniversity of North Texas, College of Arts and SciencesDentonUSA
  2. 2.Waters CorporationBeverlyUSA
  3. 3.Virginia Bioinformatics InstituteVirginia Polytechnic Institute and State UniversityBlacksburgUSA

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