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High resolution multichannel fluorescence detection for capillary electrophoresis. Application to multicomponent
K E Oldenburg1, X Xi, J V Sweedler
1Department of Chemistry, Beckman Institute, University of Illinois at Urbana-Champaign 61801, USA.
Journal of Chromatography. A
|January 7, 1998
Summary
A new wavelength-resolved fluorescence detector enhances capillary electrophoresis (CE) for sensitive biomarker analysis. This system achieves high spectral resolution, enabling precise identification of compounds like bilirubin in serum.
Area of Science:
- Analytical Chemistry
- Spectroscopy
- Biochemistry
Background:
- Capillary electrophoresis (CE) is a powerful separation technique.
- Laser-induced fluorescence (LIF) enhances detection sensitivity in CE.
- Accurate spectral information is crucial for compound identification in complex matrices.
Purpose of the Study:
- To develop and characterize a novel wavelength-resolved fluorescence detector for CE.
- To improve the sensitivity and specificity of post-column fluorescence detection.
- To demonstrate the system's utility in analyzing clinically relevant analytes.
Main Methods:
- Utilized a charge injection device (CID) array detector for wavelength-resolved detection.
- Implemented a sheath flow cell for post-column fluorescence detection.
- Developed custom methods for array readout, data processing, and electropherogram generation.
Main Results:
- Achieved a limit of detection of 4.8 x 10(-11) M for fluorescein (29,000 molecules).
- Obtained a spectral resolution of 0.56 nm/pixel, monitoring a 250 nm spectrum range (250-875 nm).
- Successfully applied the system to characterize bilirubins in human serum, matching peaks to standards via spectral data.
Conclusions:
- The developed detector provides sensitive and spectrally resolved LIF detection in CE.
- The system enables accurate identification and characterization of analytes based on spectral signatures.
- This technology offers a valuable tool for clinical diagnostics and biochemical analysis.