Simultaneous Quantitation of Multiple Biological Thiols Using Reactive Ionization and Derivatization with Charged...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Biomedical Science

Background:

• Biologically significant thiols are crucial for redox homeostasis.
• Clinical settings require rapid, accurate, simultaneous measurement of these thiols in complex biofluids at low concentrations.
• Existing methods may lack the speed, sensitivity, or simultaneous detection capabilities needed.

Purpose of the Study:

• To develop a novel derivatizing reagent and method for sensitive, simultaneous quantification of key biological thiols.
• To enable rapid analysis of thiols in various biological matrices.
• To establish a validated method for monitoring thiol redox homeostasis.

Main Methods:

• Development of a new derivatizing reagent combining thiol-selectivity and mass spectrometry sensitivity.
• Simultaneous derivatization and ionization ('reactive ionization') coupled with mass spectrometry.
• Application to diverse biofluid and tissue samples, including a cystine-cysteine redox assay.

Main Results:

• The method demonstrates high sensitivity across a wide concentration range (1 μM to 10 mM).
• Analysis is rapid, with minimal sample preparation (<4 min) and short run times (<1 min).
• High precision and accuracy (better than 8%) were validated against HPLC-MS, including redox measurements.

Conclusions:

• The developed reactive ionization mass spectrometry method offers a significant advancement for thiol analysis in clinical and research settings.
• This technique provides a rapid, accurate, and sensitive tool for assessing redox homeostasis.
• The method's applicability to various biological samples highlights its potential for broad clinical utility.