Sensitive fluorometric determination of glutathione using fluorescent polymer dots and the dopamine-melanin nanosystem

Abstract

A bioinspired fluorometric method has been developed for the detection of glutathione (GSH) in biological fluids. It is based on the use of near-infrared fluorescent semiconducting polymer dots (P-dots) and of the dopamine (DA)-melanin nanosystem. The P-dots were prepared from poly(styrene-co-maleic anhydride), the semiconducting polymer poly[(9,9′-dioctyl-2,7-divinylenefluorenylene)-alt-2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene] and the fluorescent dye tetraphenylporphyrin. They have excitation/emission maxima at 458/656 nm, and this enables measurement to be performed with low autofluorescence and scattering background. DA can self-polymerize on the surface of the P-dots to yield a poly-DA coating. This coating, at weak alkaline pH values, causes the quenching of the fluorescence of the P-dots. However, the polymerization of DA is inhibited by GSH. Hence, quenching of fluorescence is prevented. This effect was used to design a fluorometric assay for GSH that has good selectivity and sensitivity. Under optimal conditions, the method has a linear response in the 0.2 to 20 μM GSH concentration range and a 60 nM detection limit. It was successfully applied to the determination of GSH in HepG2 cells and in spiked human serum.

Schematic representation of using a NIR fluorescent P-dots and dopamine (DA)-melanin nanohybrid as a probe for glutathione (GSH) detection. The P-dots were prepared from poly(styrene-co-maleic anhydride) (PSMA), the semiconducting polymer poly[(9,9′-dioctyl-2,7-divinylenefluorenylene)-alt-2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene] (PEPV) and the fluorescent dye tetraphenylporphyrin (TPP).The GSH can inhibit the dopamine self-polymerization and prevented the formation of PDA and fluorescence quenching of P-dots.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 21605021 and 21705022), Joint Funds for the Innovation of Science and Technology of Fujian province (Grant No. 2016Y9060 and 2017Y9115), the Young and Middle-aged Talent Training Project of Fujian Provincial Health and Family Planning Commission (Grant No. 2018-ZQN-75), the China Postdoctoral Science Foundation, the Natural Science Foundation of Fujian Province of China (Grant No. 2016 J05206), the Medical Innovation grant of Fujian province (Grant No. 2018-CX-49) and the Startup Fund of Mengchao Hepatobiliary Hospital of Fujian Medical University (Grant No. QDZJ-2017-004).

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Wang, J., Zheng, C., Tan, X. et al. Sensitive fluorometric determination of glutathione using fluorescent polymer dots and the dopamine-melanin nanosystem. Microchim Acta 186, 568 (2019). https://doi.org/10.1007/s00604-019-3675-3

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Keywords

  • Polydopamine
  • Semiconducting polymer dots
  • Antiquenching
  • Antioxidant
  • PEPV
  • Biothiols
  • Tetraphenylporphyrin
  • Cell extracts
  • Serum
  • Nanoparticles