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Sensitive analysis of N-blocked amino acids using high-performance liquid chromatography with paired ion electrospray ionization mass spectrometry

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Abstract

In this study, a paired ion electrospray ionization (PIESI) mass spectrometry method was developed for sensitive detection of 9-fluorenylmethyl chloroformate (Fmoc)-derivatized amino acids. The structure-optimized ion-pairing reagent was introduced post column to form positively charged complexes which can be detected in the positive ion mode. These complexes are more surface-active than the original analytes, and meanwhile, the intensity of sodium adducts was significantly reduced. The limit of detection of the amino acids obtained with the optimal ion-pairing reagent was 0.5 to 20 pg which was 5–100 times lower than the negative mode. In addition, two mass spectrometry platforms—linear ion trap and triple quadrupole—were used to compare the PIESI improvements. Eventually, the method was applied to successfully detect the level of amino acids in human urine samples with high accuracy and the added benefit of minimizing matrix effects.

A HPLC-ESI-MS/MS method by using ion-pairing reagents for sensitive detection of Fmoc amino acids

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Acknowledgements

The authors would like to thank the Robert A. Welch Foundation (Y-0026) for its financial support.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX, 76019, USA

    Yadi Wang, Siqi Du & Daniel W. Armstrong

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  1. Yadi Wang
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  2. Siqi Du
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  3. Daniel W. Armstrong
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Corresponding author

Correspondence to Daniel W. Armstrong.

Ethics declarations

Urine samples were collected at the University of Texas at Arlington and obtained with informed consent. This study was approved by the Institutional Review Board at University of Texas at Arlington, and the experiments were performed in accordance with the ethical standards.

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The authors declare that they have no conflict of interest.

Additional information

Published in the topical collection Ionic Liquids as Tunable Materials in (Bio)Analytical Chemistry with guest editors Jared L. Anderson and Kevin D. Clark.

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Wang, Y., Du, S. & Armstrong, D.W. Sensitive analysis of N-blocked amino acids using high-performance liquid chromatography with paired ion electrospray ionization mass spectrometry. Anal Bioanal Chem 410, 4725–4735 (2018). https://doi.org/10.1007/s00216-018-0901-5

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  • DOI: https://doi.org/10.1007/s00216-018-0901-5

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