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Solid-state UV laser-induced fluorescence detection in capillary electrophoresis
K C Chan1, G M Muschik, H J Issaq
1Analytical Chemistry Laboratory, SAIC Frederick, NCI-Frederick Cancer Research and Development Center, MD 21702, USA. chan@mail.ncifcrf.gov
Electrophoresis
|July 6, 2000
Summary
Compact solid-state UV lasers enable sensitive laser-induced fluorescence (LIF) detection for peptides and amino acids. This cost-effective method achieves nanomolar sensitivity for native tryptophan compounds and fluorescamine-labeled analytes.
Area of Science:
- Analytical Chemistry
- Spectroscopy
- Biochemistry
Background:
- Laser-induced fluorescence (LIF) is a sensitive detection technique.
- Capillary electrophoresis (CE) offers efficient separation of biomolecules.
- Existing LIF systems can be expensive and bulky.
Purpose of the Study:
- To evaluate the utility of compact, inexpensive solid-state UV lasers for LIF detection in CE.
- To demonstrate the sensitivity and applicability of this system for various compounds.
Main Methods:
- Utilized two solid-state UV lasers (266 nm and 355 nm) for LIF detection.
- Separated compounds using capillary electrophoresis.
- Analyzed native tryptophan-containing proteins/peptides and fluorescamine-labeled amino acids/peptides.
Main Results:
- Achieved nanomolar (nM) level detection for native tryptophan compounds and indoles at 266 nm.
- Enabled nM level detection for fluorescamine-labeled amino acids and peptides at 355 nm.
- Demonstrated detection of amino acids as low as 10 ng/mL after fluorescamine labeling.
Conclusions:
- Solid-state UV lasers provide a stable, sufficient power source for sensitive LIF detection.
- This approach offers a cost-effective and compact alternative for biomolecule analysis.
- The method is effective for both native fluorescence and derivatized analytes, including tryptic digests.