A highly selective copper(II) ion fluorescent sensor has been designed based on the UV-visible absorption of a spiropyran derivative coupled with the use of a metal porphyrin operative on the fluorescence inner filter effect. Spiropyrans, which combine the characteristics of metal binding and signal transduction, have been widely utilized in cationic ion recognition by UV-visible spectroscopy. In the present work, the viability of converting the absorption signal of the spiropyran molecule into a fluorescence signal was explored. On account of overlap of the absorption band of the spiropyran (lambda(abs) = 547 nm) in the presence of copper ion with the Q-band of an added fluorophore, zinc meso-tetraphenylporphyrin (lambda(abs) = 556 nm), the effective light absorbed by the porphyrin and concomitantly the emitted light intensity vary as a result of varying absorption of the spiropyran via fluorescence inner filter effect. The metal binding characteristic of the spiropyran presents an excellent selectivity for copper ion in comparison with several other heavy or transition metal ions. Since the changes in the absorbance of the absorber translate into exponential changes in fluorescence of the fluorophore, the novelty of the present device is that the analytical signal is more sensitive over that of the absorptiometry or that of the fluorometry using one single dye. To realize a practical fluorescent sensor, both the absorber and fluorophore were immobilized in a plasticized poly(vinyl chloride) membrane, and the sensing characteristics of the membrane for copper ion were investigated. The sensor is useful for measuring Cu2+ at concentrations ranging from 7.5 x 10(-7) to 3.6 x 10(-5) M with a detection limit of 1.5 x 10(-7) M. The sensor is chemically reversible, the fluorescence was switched off by immersing the membrane in copper ion solution and switched on by washing it with EDTA solution.

The resonance light-scattering technique, using a spectrofluorometer, was first developed as a sensitive instrumental analysis method. At pH 7.48 and ionic strength 0.004, the extent of light-scattering of alpha, beta, gamma, delta-tetrakis[4-(trimethylammoniumyl)phenyl]porphine (TAPP) is enhanced by nucleic acids near 432 nm. There are linear relationships between the enhanced extents of light-scattering and the concentrations of nucleic acids in the range of 1.8 x 10(-7)-10.8 x 10(-7) M for calf thymus and fish sperm DNA and in the range of 1.8 x 10(-7)-1.8 x 10(-6) M for yeast RNA. The limit of determination (3 sigma) is 4.1 x 10(-8) M for calf thymus DNA, 4.6 x 10(-8) M for fish sperm DNA, and 6.7 x 10(-8) M for yeast RNA. Mechanism study indicates that nucleic acids react with the title porphyrin in two modes, depending on the concentrations of nucleic acids. When the molar ratio of nucleic acids to TAPP is smaller than 4:1, the hypochromicity and fluorescence quenching of TAPP by nucleic acids appear, and the enhancement of resonance light-scattering can be observed. When the molar ratio of nucleic acids to TAPP is larger than 4:1, a new fluorescent complex is formed.

Using a common spectrofluorometer to measure the intensity of resonance light-scattering, a method for determination of nucleic acids in the nanogram range has been developed. In the pH range 11.5-12.0, the resonance light-scattering of the binary complex of cobalt(II)/ 4-[(5-chloro-2-pyridyl)azo]-1,3-diaminobenzene (5-Cl-PADAB) is greatly enhanced by nucleic acids, with the maximum scattering peak located at 547.0 nm. The enhanced intensity of resonance light-scattering is in proportion to the concentration of calf thymus DNA in the range 0-400 ng/mL and to that of fish sperm DNA and yeast RNA in the range 0-300 ng/mL. The limits of detection are 1.4 ng/mL for calf thymus DNA, 0.8 ng/ mL for fish sperm DNA, and 1.3 ng/mL for yeast RNA. Precision at 200 ng/mL for the three nucleic acids is 1.9%, 2.0%, and 0.8%, respectively. Six synthetic samples were determined satisfactorily. Mechanism studies showed that the nature of the reaction is that the binary complex of Co(II)/5-Cl -PADAB reacts with single-stranded nucleic acid, and the enhancement effect of nucleic acids on the resonance light scattering of the binary complex is due to the stacking of the binary complex on nucleic acids, which act as a template.

The protonation of a simple meso-tetraphenylporphyrin in an organic-aqueous system was found to be induced by the counteranions. During the process of protonation, the counteranion of the proton sources binds with the porphyrin core and thus promotes the complexation of the porphyrin and protons. The interaction of porphyrin and anion was characterized by fluorescence, UV-visible, cyclic voltammetry, (1)H NMR, and IR. Moreover, it could be exploited in selective fluorescent sensing of Cl(-). The sensing mechanism was based on extraction of protons from the aqueous phase into the organic phase by free base porphyrin and simultaneous coextraction of Cl(-), which promoted porphyrin protonation, and hence resulted in significant changes of the porphyrin fluorescence spectra. Selectivity trends turned out to be dependent upon the lipophilicity of anion and the binding affinity and structure complementarity between the protonated porphyrin and anions. The fluorescence enhancement of the porphyrin band at 684 nm showed modest selectivity for Cl(-) and NO(3)(-).