Analytical and Bioanalytical Chemistry

, Volume 397, Issue 1, pp 223–231 | Cite as

Determination of heterocyclic amines in urine samples by capillary liquid chromatography with evaporated light-scattering detection

  • Fernando De Andrés
  • Mohammed Zougagh
  • Gregorio Castañeda
  • Angel Ríos
Original Paper


A rapid and simple method for separation and detection of six heterocyclic aromatic amines (2-amino-1-methyl-6-phenylimidazo [4,5-b]-pyridine, 2-amino-1-methyl-imidazo [4,5-f]-quinoline, 2-amino-3,8-dimethyl-imidazo [4,5-f]-quinoxaline, 2-amino-3,7,8-trimethyl-imidazo [4,5-f]-quinoxaline, 2-amino-3,4,8-trimethyl-imidazo [4,5-f]-quinoxaline, and 2-amino-3,4-dimethyl-imidazo [4,5-f]-quinoline) in human urine samples is proposed to reflect daily intake and recent HAAs exposure. This method comprises previous clean-up and preconcentration of the analytes on Strata-X reversed phase extraction cartridges followed by capillary liquid chromatography (CLC) and evaporative light-scattering detection (ELSD). A mobile phase of acetonitrile and ammonium acetate 35 mM at pH 5.15 through a gradient of composition and a flow rate of 15 μL min−1 resulted in good separations of the analytes. Temperature and gas pressure were optimized for detection. The CLC-ELSD allows the separation and quantification of HAAs with good resolution, precision, and sensitivity. The usefulness of the proposed method was demonstrated by the analysis of synthetic and natural human urine samples spiked with different concentration levels of heterocyclic amines.


Capillary liquid chromatography–evaporative light-scattering detection Strata-X cartridges Heterocyclic aromatic amines Human urine samples 



The Spanish Ministry of Science and Innovation (MICINN) and JJCC Castilla-La Mancha are gratefully acknowledged for funding this work with Grants CTQ2007-61830 and PCC08-0015-0722, respectively. The support given through a “INCRECYT” research contract to M. Zougagh is also acknowledged.


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Fernando De Andrés
    • 1
  • Mohammed Zougagh
    • 1
    • 2
  • Gregorio Castañeda
    • 1
  • Angel Ríos
    • 1
    • 3
  1. 1.Department of Analytical Chemistry and Food Technology, Faculty of ChemistryUniversity of Castilla-La ManchaCiudad RealSpain
  2. 2.Albacete Science and Technology ParkAlbaceteSpain
  3. 3.Regional Institute for Applied Chemistry ResearchIRICACiudad RealSpain

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