Fluorescent Probes and Mass Spectrometry-Based Methods to Quantify Thiols in Biological Systems

Area of Science:

• Redox biology
• Analytical chemistry
• Biochemistry

Background:

• Thiols are crucial sulfur-containing molecules in biological systems, impacting physiology and pathology.
• Quantifying thiol species like cysteine and glutathione is vital for understanding their biological roles.
• Fluorescent probes and mass spectrometry are key complementary methods for thiol analysis.

Purpose of the Study:

• To provide a guide for redox biologists on available methods for thiol quantification.
• To review fluorescent probes and mass spectrometry approaches for detecting thiols in biological systems.
• To highlight the complementary nature of fluorescent probes and mass spectrometry in thiol analysis.

Main Methods:

• Focus on widely used and commercially available fluorescent probes for live-cell thiol detection.
• Review general approaches for mass spectrometry-based thiol quantification.
• Emphasize cross-validation between fluorescent probes and mass spectrometry for reliable results.

Main Results:

• Fluorescent probes offer spatial and temporal resolution for thiol detection in living cells.
• Mass spectrometry serves as a gold standard for metabolite quantification.
• Reliable quantification using fluorescent probes remains a challenge, necessitating validation.

Conclusions:

• Combining fluorescent probes and mass spectrometry is recommended for accurate thiol quantification.
• Future efforts should focus on developing quantitative fluorescent probes for live cells.
• Accessibility of chemical probes to biologists and integration into metabolomics workflows are crucial.