RESUMO
A laser-induced fluorescence detector for liquid chromatography was developed. This detector was assessed by utilizing it in conjunction with gel filtration chromatography. Using the 488 nm line of an argon ion laser for excitation and monitoring the emitted fluorescence centering at 535 nm, the limit of detection of fluorescein was 580 fM. Bovine serum albumin labeled with fluorescein was detected at a concentration of 500 fM.
Assuntos
Cromatografia Líquida/métodos , Lasers , Proteínas/análise , Espectrometria de Fluorescência , Eletroforese Capilar , Fluoresceína/análise , Sensibilidade e Especificidade , Soroalbumina Bovina/análiseRESUMO
beta-Galactosidase was incubated for 60 min with the fluorogenic substrate resorufin-beta-D-galactopyranoside, which is converted by the action of the enzyme into resorufin and galactose. A 160 pL aliquot of reaction mixture was analyzed by capillary electrophoresis utilizing laser-induced fluorescence detection. Based on the detection of the resorufin formed, the limit of detection of beta-galactosidase was 1.5 x 10(-15) M or 900 molecules of enzyme in a 1 microL sample.
