Solid-state UV laser-induced fluorescence detection in capillary electrophoresis.
Two solid-state UV lasers were applied to the laser-induced fluorescence (LIF) detection of various groups of compounds after separation by capillary electrophoresis. These lasers are thermoelectric-cooled, highly compact, and inexpensive. Such lasers provide few mW of quasi-continuous wave (CW) power which are sufficient and stable for LIF detection. Native fluorescence detection of tryptophan-containing proteins and peptides and related indoles was achieved at the nM level with the laser operating at 266 nm. Detection of fluorescamine-labeled amino acids and peptides was also possible at the nM level with the laser operating at 355 nm. Amino acids at a concentration as low as 10 ng/mL could be labeled with fluorescamine. Solid-state UV-LIF detection of the tryptic digest of cytochrome c after fluorescamine derivatization was demonstrated.
Chan
K C
KC
Analytical Chemistry Laboratory, SAIC Frederick, NCI-Frederick Cancer Research and Development Center, MD 21702, USA. chan@mail.ncifcrf.gov
Muschik
G M
GM
Issaq
H J
HJ
D000596:Amino Acids / Q000032:analysis* / Q000302:isolation & purification; D000804:Angiotensin II / Q000032:analysis / Q000302:isolation & purification; D003574:Cytochrome c Group / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D005450:Fluorescamine; D005456:Fluorescent Dyes; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification; D010455:Peptides / Q000032:analysis* / Q000302:isolation & purification; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods; D014357:Trypsin
D000596:Amino Acids; D003574:Cytochrome c Group; D005456:Fluorescent Dyes; D010446:Peptide Fragments; D010455:Peptides; D011506:Proteins; D000804:Angiotensin II; D005450:Fluorescamine; D014357:Trypsin