HPLC Study of Product Formed in the Reaction of NBD-Derived Fluorescent Probe with Hydrogen Sulfide, Cysteine, N-acetylcysteine, and Glutathione

Molecules. 2022 Nov 28;27(23):8305. doi: 10.3390/molecules27238305.

Abstract

Hydrogen sulfide (H2S) and its bioderivatives analogs, such as L-cysteine (L-Cys) and glutathione (GSH), are ubiquitous biological thiols in the physiological and pathological processes of living systems. Their aberrant concentration levels are associated with many diseases. Although several NBD-based fluorescence probes have been developed to detect biological thiols, the HPLC-detection of H2S, GSH, L-Cys, and N-acetylcysteine-specific products has not been described. Herein, a novel NBD-derived pro-coumarin probe has been synthesized and used to develop a new strategy for the triple mode detection of H2S and such thiols as GSH, L-Cys, and NAC. Hydrogen sulfide and those biothiols at physiological pH release fluorescent coumarin from the probe and cause a significant fluorescence enhancement at 473 nm. The appropriate NBD-derived product for H2S, L-Cys, GSH, and NAC has a different color and retention time that allows distinguishing these biological thiols meaning the probe has a great possibility in the biological application. Fluorescent imaging combined with colorimetric and HPLC detection of H2S/biothiol-specific product(s) brings a potential tool for confirming the presence of biological thiols and determining concentrations in various aqueous biological samples.

Keywords: biothiols detections; colorimetric probe; coumarin-derived probe; fluorescent probe; hydrogen sulfide; markers.

MeSH terms

  • Acetylcysteine
  • Cysteine*
  • Fluorescent Dyes
  • Glutathione
  • HeLa Cells
  • Homocysteine
  • Humans
  • Hydrogen Sulfide*
  • Optical Imaging
  • Sulfhydryl Compounds

Substances

  • Cysteine
  • Fluorescent Dyes
  • Hydrogen Sulfide
  • Acetylcysteine
  • Glutathione
  • Sulfhydryl Compounds
  • Homocysteine