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Optimal Conditions for the Direct RP-HPLC Determination of Underivatized Amino Acids with Online Multiple Detection

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Amino Acid Analysis

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

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Abstract

The combined use of a dual-UV detector as well as a fluorimetric and a multielectrode electrochemical detector (equipped with a dual electrode, consisting of a conventional size 3 mm diameter glassy carbon electrode (GCE) and of a pair of 30 μm thick carbon microfibers) is proposed for the detection of the following 15 underivatized amino acids: l-histidine (His), l-cysteine (Cys), creatine (Crn), S-methyl-l-cysteine (Me-Cys), dl-homocysteine (Hcy), l-methionine (Met), beta-(3,4-dihydroxyphenyl)-l-alanine (DOPA), l-tyrosine (Tyr), dl-m-tyrosine (m-Tyr), l-a-methyl-DOPA (Me-DOPA), l-phenylalanine (Phe), dl-alpha-methyltyrosine (Me-Tyr), 5-hydroxy-tryptophan (5-HTP), 3-nitro-l-tyrosine (NO2Tyr), and l-tryptophan (Trp), as well as of 2 dipeptides l-cystathionine (Cysta) and l-carnosine (Car), and of creatinine (Cre). A multilinear solvent (acetonitrile) gradient elution program, determined by a simple optimization algorithm, is required for the efficient reversed-phase separation of the above mixture of 18 solutes within 27 min at a flow rate of 1.0 mL/min and at 25 °C.

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Acknowledgment

This work was supported by European Union/European Social Fund (PhD scholarship, Hrakleitos I).

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Authors and Affiliations

  1. Department of Chemistry, Laboratory of Physical Chemistry, Aristotle University of Thessaloniki, Thessaloniki, Greece

    A. Pappa-Louisi, P. Agrafiotou & S. Sotiropoulos

Authors
  1. A. Pappa-Louisi
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  2. P. Agrafiotou
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  3. S. Sotiropoulos
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Corresponding author

Correspondence to A. Pappa-Louisi .

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Editors and Affiliations

  1. Office of Policy for Pharmaceutical Quality Office of Pharmaceutical Quality, Center for Drug Evaluation and Research US FDA, Silver Spring, MD, USA

    Michail A. Alterman

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Pappa-Louisi, A., Agrafiotou, P., Sotiropoulos, S. (2019). Optimal Conditions for the Direct RP-HPLC Determination of Underivatized Amino Acids with Online Multiple Detection. 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_31

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  • DOI: https://doi.org/10.1007/978-1-4939-9639-1_31

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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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