Fluorescent RET-Based Chemosensor Bearing 1,8-Naphthalimide and Styrylpyridine Chromophores for Ratiometric Detection of Hg(2+) and Its Bio-Application

Dyad compound NI-SP bearing 1,8-naphthalimide (NI) and styrylpyridine (SP) photoactive units, in which the N-phenylazadithia-15-crown-5 ether receptor is linked with the energy donor naphthalimide chromophore, has been evaluated as a ratiometric fluorescent chemosensor for mercury (II) ions in living cells. In an aqueous solution, NI-SP selectively responds to the presence of Hg(2+) via the enhancement in the emission intensity of NI due to the inhibition of the photoinduced electron transfer from the receptor to the NI fragment. At the same time, the long wavelength fluorescence band of SP, arising as a result of resonance energy transfer from the excited NI unit, appears to be virtually unchanged upon Hg(2+) binding. This allows self-calibration of the optical response. The observed spectral behavior is consistent with the formation of the (NI-SP)·Hg(2+) complex (dissociation constant 0.13 ± 0.04 µM). Bio-imaging studies showed that the ratio of fluorescence intensity in the 440–510 nm spectral region to that in the 590–650 nm region increases from 1.1 to 2.8 when cells are exposed to an increasing concentration of mercury (II) ions, thus enabling the detection of intracellular Hg(2+) ions and their quantitative analysis in the 0.04–1.65 μM concentration range.