New reagents for determination of amino acids by liquid chromatography with pre-column fluorescence derivatization

Xinjun Fan, Jinmao You, Jingwu Kang, Qingyu Ou, Qingcun Zhu

Research output: Contribution to journalArticle

45 Scopus citations

Abstract

A liquid-chromatography method for the determination of amino acids with fluorescence detection has been developed. The reactions of three fluorescent tagging reagents, namely acridone-N-acetyl chloride (ARC-Cl), carbazole-9-acetyl chloride (CRA-Cl) and carbazole-9-propionyl chloride (CRP-Cl), with amino acids are reported. Emission maxima for the ARC, CRA and CRP derivatives of amino acids were at 430nm (λ(ex)=404nm), 360nm (λ(ex)=335nm) and 365nm (λ(ex)=340nm), respectively. In all cases, the derivatives exhibited strong fluorescence, whereas the reagents themselves also exhibited fluorescence. The labeled derivatives are very stable; <4% decomposition occurs after heating at 40°C for 24h. The fluorescence intensities of the amino-acid derivatives were higher in neutral and alkaline than in acidic solutions. This method, combining a multi-gradient program, offers baseline resolution of the common ARC, CRA and CRP amino-acid derivatives than a linear acetonitrile gradient. Separation of amino-acid derivatives was carried out on a reversed-phase C18 column. Derivatization and chromatographic conditions were optimized. The separation of amino-acid derivatives has good reproducibility and the relative standard deviations (n=6) for 5pmol of each amino acid are <4%. The detection limit is at the fmol level. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)81-91
Number of pages11
JournalAnalytica Chimica Acta
Volume367
Issue number1-3
DOIs
Publication statusPublished - Jul 3 1998
Externally publishedYes

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Keywords

  • Acridone-N-acetyl chloride
  • Amino acids
  • Carbazole-9-acetyl chloride
  • Carbazole-9-yl-propionyl chloride
  • Fluorescence derivatization
  • Liquid chromatography

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

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