The rapid synthesis of a fluorescent probe core for hydrolytic enzymes
Date Issued
2007
Date
2007
Author(s)
Liu, Tzu-Chen
Abstract
A ratiometric measurement, i.e. simultaneously recording of the fluorescence intensities at two wavelengths and calculation of their ratio, is a technique that can provide precise data and even quantitative detection. To carry out this idea, the probe must exhibit a large shift in its emission or excitation spectrum after it reacts with the target molecule. A strategy that is based on fluorescence resonance energy transfer (FRET) by using a two-fluorophore cassette comprised of a coumarin donor, a cyclohexane linker and a fluorescein acceptor has been utilized in many fluorescent sensors. However, with this structural similarity, all of the previous works were done by synthesizing from the first step to the last step, which is wasteful and tedious. Therefore, in order to establish a more efficient model, in this study, our work is separated into two parts. First, we focus on developing a fast procedure of synthesizing fluorescent probe core. Second, we utilize the two hydroxyl groups on the phenols to link with different functional groups. The conclusion shows the results to be practical as the hydroxyl group is chemically reactive and is able to link with fatty acid, sulfate and the protecting group of sulfate. Further fluorescence spectra also confirm that the emission wavelength of the fluorescent probe core differs by around 40 nm before/after the conjugation with functional groups. The value of this work is the establishment of a platform suitable for detecting various hydrolytic enzymes, and during the process of this study, we discovered a method to separate the regioisomers of carboxyfluorescien via fractional recrystallization in a double-digit gram scale.
Subjects
core
fluorescent probes
fluorescence resonance energy transfer
ratiometric measurement
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