Simultaneous determination of eight biologically active thiol compounds using gradient elution-liquid chromatography...

Area of Science:

• Biochemistry
• Analytical Chemistry

Background:

• The thiol group is a crucial functional group in many biologically active molecules.
• Accurate quantification of thiols is essential for understanding various biological processes.

Purpose of the Study:

• To develop and optimize a flow injection analysis/high-performance liquid chromatography (FIA/HPLC) method with electrochemical detection for the simultaneous determination of multiple thiol compounds.
• To establish sensitive detection limits for various thiols in biological matrices.

Main Methods:

• Optimization of electrochemical detection potentials, mobile phase organic solvent content, and elution profiles (isocratic and gradient).
• Utilized HPLC with electrochemical detection for separation and quantification of cysteine, cystine, N-acetylcysteine, homocysteine, glutathione (reduced and oxidized), desglycinephytochelatin, and phytochelatins.
• Evaluated the impact of gradient elution profiles and mobile phase composition on thiol separation.

Main Results:

• Achieved well-separated detection of multiple thiol compounds using optimized HPLC-electrochemical detection conditions.
• Established high sensitivity with detection limits in the tens of femtomoles per injection for most thiols, except phytochelatin5 (2.1 pmol).
• Successfully applied the developed method for simultaneous thiol determination in biological samples, including maize tissue and human blood serum.

Conclusions:

• The developed FIA/HPLC-electrochemical detection method provides a sensitive and robust approach for simultaneous thiol analysis.
• This technique is suitable for quantifying biologically relevant thiols in complex biological samples.