High performance liquid chromatography coupled to an optical fiber detector coated with laccase for screening catecholamines in plasma and urine

Filipe D.P. Ferreira, Lurdes I.B. Silva*, A. C. Freitas, Teresa A. P. Rocha-Santos, A. C. Duarte

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

43 Citations (Scopus)

Abstract

An analytical method based on separation by high performance liquid chromatography (HPLC) and detection by optical fiber (OF) coated with an enzyme (laccase), has been developed for separation and quantification of catecholamines, namely epinephrine, dopamine and norepinephrine. The application of OF as a detector in this analytical system relies on the variation of the reflected optical power detected when the catecholamines eluted from the HPLC column act as the substrate of the laccase immobilized on a tip of a single-mode OF. The developed method shows a high linearity in a range between 5 and 125 pg/mL and detection limits of 3.5, 2.9 and 3.3 pg/mL for epinephrine, dopamine and norepinephrine, respectively. The analytical performance of the proposed method was compared with a classical analytical method, namely high performance liquid chromatography-electrochemical detector (HPLC-ED) regarding catecholamines detection, showing great analytical advantages such as low cost of equipment. Additionally, the proposed method was applied to catecholamines determination in actual samples of plasma and human urine.
Original languageEnglish
Pages (from-to)7049-7054
Number of pages6
JournalJournal of Chromatography A
Volume1216
Issue number42
DOIs
Publication statusPublished - 16 Oct 2009
Externally publishedYes

Keywords

  • Catecholamines
  • HPLC
  • Laccase
  • Optical fiber detector
  • Plasma
  • Urine

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