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General contribution of nonspecific interactions to fluorescence intensity

AuthorsGálvez Buerba, Eva Mª ; Matt, Muriel; Cebolla, Vicente L. ; Fernandes, Francisco; Membrado, Luis; Cossío, Fernando P.; Garriga, Rosa; Vela, Jesús; Guermouche, M. Hassan
KeywordsFluorescence detection
Intensity changes
Issue Date28-Apr-2006
PublisherAmerican Chemical Society
CitationAnalytical Chemistry - Columbus 78(11): 3699-3705 (2006)
AbstractMany chemical compounds, including nonfluorescent ones, induce changes in the fluorescence spectra of certain probes, such as berberine cation and Reichardt's betaine, both in the absence and the presence of solvent, that affect almost exclusively emission intensity. In this work, the application of fluorescence detection by intensity changes (FDIC) to HPLC and TLC chromatographic systems with fluorescence detectors has been studied. FDIC detection is of special interest in detecting nonfluorescent analytes, either in HPLC or in TLC mode. It does not involve covalent interactions, and the dielectric permittivity (ε) of the medium plays an important role. The balance between nonspecific and specific interactions produces either an increase or a decrease in fluorescence intensity. Therefore, the influence of chromatographic conditions and chemical structure of analytes on the sign and magnitude of fluorescence peaks for sample detection in HPLC and TLC systems has been discussed. In general, probe nature and concentration determine response and detection sensitivity for a given sample in TLC and HPLC. As solubility and fluorescence properties in solvents determine the operating conditions for a FDIC probe in HPLC mode, nature and flows of mobile phase and solvent are important for chromatographic response and detection sensitivity.
Description6 Figuras.
Publisher version (URL)http://dx.doi.org/10.1021/ac058045b
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