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Light emitting diode excitation emission matrix fluorescence spectroscopy
1Naval Research Laboratory, Chemistry Division, Biological Chemistry, Code 6113, 4555 Overlook Ave, S.W, Washington, DC 20375, USA. shart@ccf.nrl.navy.mil
The Analyst
|January 23, 2003
Summary
A new excitation emission matrix (EEM) fluorescence instrument uses a low-cost LED array for multi-wavelength excitation. This system achieves high sensitivity, with detection limits in the parts-per-trillion range for some dyes.
Area of Science:
- Analytical Chemistry
- Spectroscopy
- Instrumentation
Background:
- Traditional EEM fluorescence instruments often rely on expensive high-power lamps and imaging spectrographs.
- Developing cost-effective alternatives for multi-wavelength excitation is crucial for broader accessibility.
Purpose of the Study:
- To develop and characterize a novel EEM fluorescence instrument utilizing a linear array of light-emitting diodes (LEDs).
- To evaluate the performance of the LED-EEM system for detecting and quantifying fluorescent dyes.
Main Methods:
- Construction of an EEM fluorescence instrument with an LED array covering 370-640 nm.
- Characterization using fluorescent dyes, including 9,10-bis(phenylethynyl)-anthracene and rhodamine B.
- Analysis of data using parallel factor analysis (PARAFAC) for component resolution.
Main Results:
- Achieved low limits of detection (LOD) for fluorescent dyes, in the mid parts-per-trillion range for some compounds.
- Demonstrated accurate identification and quantitation of six fluorescent dyes in mixtures with root mean squared errors of prediction (RMSEP) below 4.0 ppb.
- PARAFAC successfully resolved individual contributions of fluorescein tautomers.
Conclusions:
- The LED-EEM system offers a cost-effective and sensitive alternative to conventional EEM spectroscopy.
- The developed instrument is capable of accurate multi-component analysis of fluorescent dyes.
- This hybrid spectroscopic-sensor system shows promise for various analytical applications.