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Capillary electrophoresis with laser-induced native fluorescence detection for profiling body fluids
D M Paquette1, R Sing, P R Banks
1Department of Chemistry and Biochemistry, Concordia University, Montréal, Québec, Canada.
Journal of Chromatography. B, Biomedical Sciences and Applications
|September 24, 1998
Summary
This study demonstrates laser-induced native fluorescence for analyzing biological fluids like urine, saliva, and serum using capillary electrophoresis (CE). The method achieves sensitive detection without sample derivatization, offering a direct analytical approach.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Spectroscopy
Background:
- Capillary electrophoresis (CE) is a powerful separation technique.
- Native fluorescence detection offers a label-free analytical approach.
- Derivatization is often required to enhance fluorescence, complicating analysis.
Purpose of the Study:
- To investigate laser-induced native fluorescence detection for CE analysis of biological samples.
- To evaluate the performance of a KrF excimer laser as an excitation source.
- To determine detection limits for various analytes in human urine, saliva, and serum.
Main Methods:
- Utilized a KrF excimer laser (248 nm) for native fluorescence detection in CE.
- Separations performed in phosphate/tetraborate buffers at alkaline pH using a 50-µm I.D. capillary.
- Micellar electrokinetic chromatography (MEKC) with sodium dodecyl sulfate employed for urine analysis.
- Laser operated at high pulse repetition rates (553–2009 Hz) to simulate continuous wave excitation.
Main Results:
- Achieved sensitive detection of tryptophan (4 nM), conalbumin (10 nM), and α-lactalbumin (30 nM) in free solution CE.
- Determined detection limits for indole compounds and catecholamine metabolites in urine via MEKC ranged from 7–170 nM.
- Detection limits correlated with average laser excitation power, influenced by pulse rate.
Conclusions:
- Laser-induced native fluorescence is a viable, sensitive detection method for CE of biological samples.
- The technique eliminates the need for sample derivatization, simplifying sample preparation.
- KrF excimer laser excitation provides effective native fluorescence detection for complex biological matrices.