A simple liquid chromatographic method based on intramolecular excimer-forming derivatization and fluorescence detection for the determination of tyrosine and tyramine in urine

J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Jul 5;821(1):88-93. doi: 10.1016/j.jchromb.2005.04.004.

Abstract

A liquid chromatographic (LC) method for sensitive and selective fluorometric determination of p-hydroxyphenylethylamino group containing compounds is described. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(1-pyrene)butanoyl chloride, followed by reversed-phase LC. The analytes, containing an amino moiety and a phenolic hydroxyl moiety in a molecule, were converted to the corresponding dipyrene-labeled derivatives by one-step derivatization. The dipyrene-labeled derivatives afforded intramolecular excimer fluorescence (440-540 nm), which can clearly be discriminated from the normal fluorescence (360-420 nm) emitted from reagent blanks. The derivatives of tyrosine and tyramine could be separated by reversed-phase LC on ODS column under conditions of isocratic elution. The detection limits (signal-to-noise ratio = 3) for tyrosine and tyramine were 4.5 and 2.6 fmol per 20 microL injection, which corresponded to analyte concentrations of 0.9 and 0.5 nM, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Butanes / chemistry
  • Chromatography, Liquid / methods*
  • Humans
  • Pyrenes / chemistry
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence
  • Tyramine / urine*
  • Tyrosine / urine*

Substances

  • 4-(1-pyrene)butanoyl chloride
  • Butanes
  • Pyrenes
  • Tyrosine
  • Tyramine