Selective and sensitive liquid chromatographic determination method of 5-hydroxyindoles with fluorous and fluorogenic...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Pharmacology

Background:

• 5-hydroxyindoles (5-HIs) are crucial biomarkers.
• Accurate quantification of 5-HIs is essential for clinical diagnostics.
• Existing methods for 5-HI analysis often lack selectivity and sensitivity.

Purpose of the Study:

• To develop a highly selective and sensitive liquid chromatography (LC) method for simultaneous determination of 5-hydroxyindoles.
• To establish a novel analytical approach for 5-HI quantification in biological samples.

Main Methods:

• Precolumn derivatization of 5-HIs using 4-(3',3',4',4',5',5',6',6',7',7',8',8',9',9',10',10',10'-heptadecafluorodecyl)benzylamine (HFBA).
• Separation of HFBA-derivatized 5-HIs on a fluorous liquid chromatography (LC) column.
• Detection of derivatized 5-HIs utilizing their enhanced fluorescent properties.

Main Results:

• Stable benzoxazole derivatives of 5-HIs were formed via a straightforward derivatization process.
• Selective retention of HFBA-5-HI derivatives on the fluorous LC column was achieved through fluorous interactions.
• The method provided sensitive detection with limits of 1.2-14 fmol for 5-HIs.
• Analysis of 5-HIs in human plasma from healthy subjects was successfully performed.

Conclusions:

• The developed fluorous LC method offers improved selectivity and sensitivity for simultaneous 5-HI determination.
• This method is suitable for analyzing 5-hydroxyindoles in complex biological matrices like human plasma.
• The technique provides a valuable tool for research and clinical applications involving 5-HI metabolism.