Novel Fluorescent/EPR Difunctional Probe for Detecting Hypochlorite†

doi:10.7503/cjcu20170193

Abstract

Abstract:

To detect hypochlorite effectively and multimodally in biological system, a novel fluorescence/EPR dual probe CNNOH was designed and synthesized based on coumarin and naphthalimide derivatives. The phy-sicochemical properties of CNNOH and its reaction with hypochlorite were studied in detail by fluorescence, electron paramagnetic resonance(EPR) and ultraviolet-visible spectroscopies. The results showed that hypochlorite can be detected using CNNOH by fluorescence and EPR spectroscopies with integrated advantages of both techniques. Owing to fluorescence resonance energy transfer(FRET) effect between the two fluorophores, the probe exhibits a large Stokes shift(135 nm) and thus effectively eliminates interference from of stray light. This dual probe has multiple advantages, such as low detection limit(0.214 μmol/L), short response time to hypochlorite(ca. 10 s), measurement of hypochlorite over a wide range of concentrations(0—5 mmol/L) as well as good selectivity and stability of CNNOH under physiological conditions. Therefore, the new probe shows great potentials in detecting or imaging hypochlorite in living cells and this molecular design provides new insights into the development of new probes for hypochlorite.

Key words: Hypochlorite, Fluorescent probe, Electron paramagnetic resonance(EPR), Fluorescence resonance energy transfer(FRET), Free radical

CLC Number:

O657

TrendMD:

LIU Huiqiang, PENG Chao, CHEN Ning, LIU Yangping. Novel Fluorescent/EPR Difunctional Probe for Detecting Hypochlorite^†[J]. Chem. J. Chinese Universities, 2017, 38(9): 1542.

Figures/Tables 6

Scheme 1 Synthetic route of CNNOH

Fig.1 Absorption spectra of 10 μmol/L of CNNOH(a), compound 4(b) or compound 5(c) in PBS buffer(A), fluorescence spectra of 1 μmol/L of CNNOH(a), compound 4(b) or compound 5(c)(B) and overlap of fluorescence spectra(a) of compound 5(donor, 10 μmol/L) and absorption spectra(b) of compound 4(acceptor, 1 μmol/L)(C)

Fig.2 Fluorescence intensity changes of CNNOH(1 μmol/L) in the presence of 10 μmol/L(a) or 50 μmol/L(b) ClO-

Fig.3 Spectra changes of CNNOH upon addition of ClO-(A) Absorption spectra of CNNOH(10 μmol/L) with the reaction of ClO-(0—1 mmol/L); (B) fluorescence spectra of CNNOH(1 μmol/L) with the reaction of ClO-(0—100 μmol/L); (C) EPR spectra of CNNOH(50 μmol/L) with the reaction of ClO-(0, 0.5, 2.5, 5 mmol/L); (D) fitting curve of EPR intensity with different concentrations of ClO-(0, 0.5, 1, 2, 3, 4, 5 mmol/L).

Fig.4 Fluorescence intensity of CNNOH after reaction with various ROS

Fig.5 Effects of pH value on stabilities of CNNOH(a) and its product with ClO-(b)

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