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Time-resolved high-performance liquid chromatography fluorescence detector using a nanosecond pulsed light source for
T Iwata1, J Koshoubu, Y Kurosu
1Department of Mechanical Engineering, Faculty of Engineering, University of Tokushima, Japan. iwata@me.tokushima-u.ac.jp
Journal of Chromatography. A
|November 24, 1999
Summary
A new time-resolved fluorescence detection (TRFD) system enhances amino compound analysis with high-performance liquid chromatography (HPLC). This method uses lanthanide ion chelates for highly selective detection, minimizing background interference.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Amino compounds are crucial in biological systems.
- Existing analytical methods for amino compounds can suffer from low selectivity.
- High-performance liquid chromatography (HPLC) is a standard separation technique.
Purpose of the Study:
- To develop and characterize a novel time-resolved fluorescence detection (TRFD) system for amino compound analysis.
- To improve selectivity and reduce background fluorescence in HPLC-based amino compound detection.
- To demonstrate the system's utility through performance tests.
Main Methods:
- Construction of a TRFD system utilizing a nanosecond pulsed xenon-arc lamp.
- Derivatization of amino compounds with isothiocyanobenzyl-EDTA (IEDTA) and lanthanide chelation.
- Post-column addition of an enhancer solution before TRFD detection.
Main Results:
- The developed TRFD system successfully detected derivatized amino compounds.
- The long fluorescence lifetime of lanthanide chelates provided high selectivity against background fluorescence.
- Fundamental performance tests confirmed the system's analytical capabilities.
Conclusions:
- The novel TRFD system offers a highly selective method for amino compound analysis coupled with HPLC.
- This approach effectively overcomes limitations of background fluorescence in traditional detection methods.
- The system demonstrates significant potential for sensitive and selective quantification of amino compounds.