CDs-Peroxyfluor Conjugation for Ratiometric Fluorescence Detection of Glucose and Shortening Its Detection Time from Reaction Dynamic Perspective

A ratiometric fluorescence probe based on the conjugation of peroxyfluor-NHS (PF) and carbon dots (CDs) was designed for selective and rapid detection of glucose. When glucose was catalytically oxidized by glucose oxidase (GOx), the product H2O2would react with colorless and non-fluorescent peroxyfluor moiety to give the colored and fluorescent fluorescein moiety which would absorb the energy of CDs emission at 450 nm due to the Förster Resonance Energy Transfer (FRET) and generate a new emission peak at 517 nm. The reaction between PF and H2O2was slow with a rate constant of about 2.7 × 10−4s−1under pseudo-first-order conditions (1 uM PF, 1 mM H2O2), which was unconducive to rapid detection. Given this, a short time detection method was proposed by studying the kinetics of the reaction between PF and H2O2. In this method, the detection time was fixed at three minutes. The linear detection of glucose could be well realized even if the reaction was partially done. As glucose concentration increased from 0.05 mM to 5 mM, the fluorescence intensity ratio (I517/I450) after 3 minutes’ reaction of CDs-PF and glucose oxidation products changed linearly from 0.269 to 1.127 with the limit of detection (LOD) of 17.19 μM. In addition, the applicability of the probe in blood glucose detection was verified.Keywords:carbon dots;ratiometric fluorescence;glucose;rapid detection;fluorescein;kinetics