We reevaluate the absolute fluorescence and phosphorescence quantum yields of standard solutions by using a novel instrument developed for measuring the absolute emission quantum yields of solutions. The instrument consists of an integrating sphere equipped with a monochromatized Xe arc lamp as the light source and a multichannel spectrometer. By using a back-thinned CCD (BT-CCD) as the detector, the sensitivity for spectral detection in both the short and long wavelength regions is greatly improved compared with that of an optical detection system that uses a conventional photodetector. Using this instrument, we reevaluate the absolute fluorescence quantum yields (Phi(f)) of some commonly used fluorescence standard solutions by taking into account the effect of reabsorption/reemission. The value of Phi(f) for 5 x 10(-3) M quinine bisulfate in 1 N H(2)SO(4) is measured to be 0.52, which is in good agreement with the value (0.508) obtained by Melhuish by using a modified Vavilov method. In contrast, the value of Phi(f) for 1.0 x 10(-5) M quinine bisulfate in 1 N H(2)SO(4), which is one of the most commonly used standards in quantum yield measurements based on the relative method, is measured to be 0.60. This value is significantly larger than Melhuish's value (0.546), which was estimated by extrapolating the value of Phi(f) for 5 x 10(-3) M quinine bisulfate solution to infinite dilution using the self-quenching constant. The fluorescence quantum yield of 9,10-diphenylanthracene in cyclohexane is measured to be 0.97. This system can also be used to determine the phosphorescence quantum yields (Phi(p)) of metal complexes that emit phosphorescence in the near-infrared region: the values of Phi(p) for [Ru(bpy)(3)](2+) (bpy = 2,2'-bipyridine) are estimated to be 0.063 in water and 0.095 in acetonitrile under deaerated conditions at 298 K, while that in aerated water, which is frequently used as a luminescent reference in biological studies, is reevaluated to be 0.040.

Effects of solvent water on the photophysical properties of a series of meta- and para-substituted anilines have been investigated by means of time-resolved fluorescence, transient absorption, and photoacoustic measurements. Some aniline derivatives exhibit extremely short fluorescence lifetime (tau(f)) and small quantum yield (Phi(f)) in water (e.g., tau(f) = 45 ps and Phi(f) = 0.0019 for m-cyanoaniline (m-ANCN) in H(2)O), which is in marked contrast with their much larger values in nonaqueous solvents (tau(f) = 7.3 ns and Phi(f) = 0.14 for m-ANCN in acetonitrile). Photoacoustic and transient absorption measurements show that the remarkable fluorescence quenching of m-ANCN in water is attributed almost exclusively to fast internal conversion. The lifetime measurements of m-ANCN in H(2)O/acetonitrile binary solvent mixtures reveal that the quenching is related to variation of hydrogen-bonding interactions between the amino group and water molecules and the conformational change of the amino group upon electronic excitation. Similar fluorescence quenching due to solvent water is also found for N-alkylated m-ANCNs. The drastic differences in the fluorescence intensity and lifetime of m-ANCNs under hydrophobic and hydrophilic environments and also the large solvent polarity dependence of the fluorescence band position suggest the possibility that they can be utilized as fluorescent probes for investigating the microenvironment of biological systems. In suspensions of human serum albumin (HSA) in water, remarkable enhancement of the fluorescence intensity and lifetime is observed for m-ANCN and its N-alkylated derivatives, demonstrating that m-ANCNs can be a candidate for novel fluorescent probe with small molecular size.

The excited-state proton transfer (ESPT) to solvent from phenol (PhOH) and cyanophenols (CNOHs) in water was studied by means of time-resolved fluorescence and photoacoustic spectroscopy. A characteristic property of PhOH and CNOHs is that the fluorescence quantum yields of the deprotonated forms are remarkably small (< or = 10(-3)) and the lifetimes are extremely short (< or = 30 ps). Time-resolved fluorescence measurements for PhOH, CNOHs, and their methoxy analogues at 298 K indicate that o- and m-cyanophenols (o- and m-CNOH) undergo rapid ESPT to the solvent water with rate constants of 6.6 x 10(10) and 2.6 x 10(10) s(-1), respectively, whereas the fluorescence properties of PhOH and p-CNOH does not exhibit clear evidence of the ESPT reaction. Photoacoustic measurements show that photoexcitation of o- and m-CNOH in water results in negative volume changes, supporting the occurrence of ESPT to produce a geminate ion pair. In contrast, the volume contractions for the PhOH and p-CNOH solutions are negligibly small, which indicates that, in these compounds, the yields of solvent-separated ion pairs resulting from the ESPT are very small. The volume change per absorbed Einstein (DeltaV(r)) for o-CNOH is obtained to be -5.0 mL Einstein(-1), which is much smaller than the estimated volume contraction per photoconverted mole (DeltaV(R)). This suggests that the geminate recombination between the ejected proton and the cyanophenolate anion occurs after rapid deactivation of the excited ion pair. In the temperature range between 275 and 323 K, the proton dissociation rates of o- and m-CNOH in H(2)O and D(2)O are slower than the solvent relaxation rates evaluated from the Debye dielectric relaxation time, indicating that the overall rate constant is determined mainly by the proton motion along the reaction coordinate.

The proton-transfer reactions to solvent from electronically excited o-, m-, and p-(trifluoromethyl)phenols (TFOHs) in water have been investigated by picosecond time-resolved fluorescence measurements. The rate constants for the proton dissociation of o-, m-, and p-TFOH are obtained to be 2.2×109, 8.6×108, and 2.5×108 s−1, respectively. On the basis of the rate constants, the effects of substituent and deuterium isotope effects on the proton-transfer reactions are revealed.