Abstract

Urinary pterins have been found as potential biomarkers in many pathophysiological conditions including inflammation, viral infections, and cancer. However, pterins determination in biological samples is difficult due to their degradation under exposure to air, light, and heat. Besides, they occur at shallow concentration levels, and thus, standard UV detectors cannot be used without additional sample preconcentration. On the other hand, ultra-sensitive laser-induced fluorescence (LIF) detection can be used since pterins exhibit native fluorescence. The main factor that limits an everyday use of LIF detectors is its high price. Here, an alternative detector, i.e., light-emitted diode induced fluorescence (LEDIF) detector, was evaluated for the determination of pterins in urine samples after capillary electrophoresis (CE) separation. An optimized method was validated in terms of linearity range, limit of detection (LOD), limit of quantification (LOQ), intra- and interday precision and accuracy, sample stability in the autosampler, and sample stability during the freezing/thawing cycle. The obtained LOD (0.1 µM) and LOQ (0.3 µM) values were three-order of magnitude lower compared to UV detector, and two orders of magnitude higher compared to previously reported house-built LIF detector. The applicability of the validated method was demonstrated in the analysis of urine samples from healthy individuals and cancer patients.

Highlights

  • IntroductionPterin and its derivatives are heterocyclic compounds based on the pteridine ring system [1,2,3,4]

  • Pterin and its derivatives are heterocyclic compounds based on the pteridine ring system [1,2,3,4].They occur naturally in almost all organisms in which they play many important roles such as coenzymes [5], inhibitors [6], sensitizers [7,8], sensors [9], pigments [10,11,12,13,14], and toxins [15]

  • capillary electrophoresis (CE)-LEDIF system was presented for the first time

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Summary

Introduction

Pterin and its derivatives are heterocyclic compounds based on the pteridine ring system [1,2,3,4]. They occur naturally in almost all organisms in which they play many important roles such as coenzymes [5], inhibitors [6], sensitizers [7,8], sensors [9], pigments [10,11,12,13,14], and toxins [15]. A solubility of pterins in water, as well as in various organic solvents, is minimal

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