Journal of Fluorescence

, Volume 29, Issue 6, pp 1445–1455 | Cite as

Phenylalanine Photoinduced Fluorescence and Characterization of the Photoproducts by LC-MS

  • Elísabet Martín-TorneroEmail author
  • Francisco J. Sierra-Tadeo
  • Isabel Durán-Merás
  • Anunciación Espinosa-Mansilla


Phenylalanine (Phe) is a direct precursor of tyrosine and several neurotransmitters. The accumulation of Phe in the brain generates serious and not recoverable pathologies in children. Early detection in newborns is fundamental to apply the appropriate therapy and avoid irreversible health problems. Although fluorescence is a sensitive and selective technique for the determination of amino acids, the fluorescent analysis of Phe is limited since it exhibits a very low fluorescence quantum yield; however, the fluorescence of Phe increases drastically under UV irradiation when a peroxide medium is used. The aim of this research was to analyze the effect of the UV-radiation on Phe aqueous-peroxide solutions and to study the influence of the chemical environment on the photoinducted fluorescence process. The nature and characteristics of the fluorescent photoproducts generated under off-line UV irradiation in hydrogen peroxide medium were achieved by high performance liquid chromatography (HPLC) using a spectrophotometer detector (DAD) coupled in series with a mass spectrometer (MS) or with a fast scan spectrofluorimetric detector (FSFD). Environmental characteristics such as pH, initial concentration of Phe, hydrogen peroxide amount and irradiation time were studied in order to establish their influence on the formation of each one of the photoproducts. As the formation of several highly fluorescent photoproducts has been confirmed, the possibility of designing a chromatographic system with a post-column on-line photoreactor is open. The measure of the total fluorescence signal generated from Phe at the optimized irradiation time, could be used for the determination, with high sensitivity, of the initial amount of Phe in aqueous media, such as human serum or environmental samples. These aspects are being studied at present. 


Phenylalanine Tyrosine Photoinduced fluorescence High performance liquid chromatography Mass spectrometry 



Authors are grateful to the Ministerio de Economía, Industría y Competitividad of Spain (Project CTQ2017-82496-P) and to the Junta de Extremadura (GR18041-Research Group FQM003, and Project IB16058), both cofinanced by the Fondo Europeo de Desarrollo Regional, for financially supporting this work. Elisabet Martín-Tornero thanks to Ministerio de Economía y Competitividad of Spain and Fondo Social Europeo for a FPI grant with reference BES-2015-075407 (Order ECC/1402/2013, 22 of July).


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Elísabet Martín-Tornero
    • 1
    • 2
    Email author
  • Francisco J. Sierra-Tadeo
    • 1
  • Isabel Durán-Merás
    • 1
    • 2
  • Anunciación Espinosa-Mansilla
    • 1
    • 2
  1. 1.Department of Analytical ChemistryUniversity of ExtremaduraBadajozSpain
  2. 2.Research Institute on Water, Climate Change & Sustainability (IACYS)University of ExtremaduraBadajozSpain

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