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Fluorescence spectra and absorption spectra of carvacrol, an active component of Chinese herbal medicines, have been studied. The ionization constant and fluorescence quantum yield of carvacrol were measured according to spectral data. Under the condition of pH < 2.0, fluorescence intensity of carvacrol increases with the increase in pH value. In the range of pH 2.0-8.0, carvacrol gives a strong and steady fluorescence with maximum excitation wavelength 278 nm and emission wavelength 306 nm. When pH > 8.0, the fluorescence intensity decreases with the increase in pH value. Ionization constant of carvacrol was measured to be pK(a) = 10.44 +/- 0.06 using a pH-absorbance method; and pK(a) = 10.40 +/- 0.04 using a pH-fluorescence method. Fluorescence intensity of carvacrol was remarkably enhanced when methanol was added into its aqueous solution. Using L-tryptophane as a reference, the fluorescence quantum yield of carvacrol aqueous solution was measured to be 0.121 at excitation wavelength 278 nm; while in a solution containing 80% methanol, the quantum yield was measured to be 0.324.

Three-dimensional fluorescence spectrum and ultraviolet absorption spectrum of Bletilla striata extraction drawn by boiling water were reported. Three fluorescence peaks, located at lambda(ex)/lambda(em) = 227/299 nm, 271/299 nm and 258/ 389 nm respectively, were observed in the three-dimensional fluorescence spectrum. Among them, the two former peaks having the same emission wavelengths were produced by one fluorescent component, while the latter peak was produced by another. Three-dimensional fluorescence spectrum is a characteristic mark of Bletilla striata, therefore it can be used in qualitative identification. The distance between emission wavelengths of the two components was 90 nm, so the fluorescence intensity of two components could be measured separately without pre-separation. For dilute solutions of Bletilla striata aqueous extraction, good linear relationships between fluorescence intensity and concentration of two components were obtained, thereby the quantitative determination of the two components may be carried out. In the range of pH 1.5 to pH 12.5, the fluorescence intensity of the two components changes with the increase in pH, indicating dissociable protons exist in the molecular structure of the two components.

Fluorescence spectra of chang shan (Dichroa febrifuga Lour) aqueous extraction were studied. In the three-dimensional fluorescence contour spectrum, three fluorescence peaks of quinazoline alkaloids, which are the active components of chang shan, were observed. The excitation wavelengths of the peaks were 235, 270 and 320 nm, respectively, and the emission wavelength of all the peaks was 430 nm. Three-dimensional fluorescence contour spectrum is the very image of fingerprint, suitable for qualitative identification of traditional Chinese medicine. In the range of pH 3 to pH 6, the fluorescence spectrum of chang shan aqueous extractions changes with the variation in pH value. The reason for this spectral change might be the protonation of N-1 in quinazolone ring of beta-dichroine (febrifugine) molecule. There is an excellent linear relationship between the fluorescence intensity and the concentration of chang shan under nearly neutral conditions, thereby a quantitative method for the determination of quinazoline alkaloids may be established.

Fluorescence spectra, ultraviolet absorption spectra, and protonation of Ciprofloxacin (CIP) at different pH values have been studied. Fluorescence quantum yield of CIP under neutral condition has been measured. In HCl medium with [H+] > 1 mol x L(-1), CIP molecules (simplified as HL) may accept three protons to exist as H4L3+ with very weak fluorescence, and its maximum fluorescence emission wavelength (lambdamax) is 456 nm. In acidic solution of pH 0 to pH 2, CIP mainly exists as H3L2+ form with lambdamax at 450 nm, and fluorescence intensity is relatively weak and increases with increasing of pH. In the range of pH 2 to pH 4, CIP mainly exists as H2L+ form with a strong fluorescence, and lambdamax is still at 450 nm. When pH>4, lambdamax gradually blue-shifts to 414 nm, fluorescence intensity slightly decreases as pH increases, and at the same time an evident change in ultraviolet absorption spectrum is observed, indicating that H2L+ has lost proton to exist as dipole ion form HL. When pH>8, the fluorescence intensity decreases until disappearance as pH increases, indicating that HL has lost proton to exist as non-fluorescence anion ion form L-. In the molecular form changing process, the maximum excitation wavelength of CIP is essentially constant at 275 nm, but the maximum emission wavelength changes obviously. In a buffer solution with pH 7.0, and using quinine bisulphate as a reference, the fluorescence quantum yield of CIP at maximum excitation wavelength 275 nm was measured to be 0.12.

Fluorescence spectra, ultraviolet spectra and protonation of Ofloxacin (OFL) at different pH values have been studied. In strong acidic solutions, OFL molecule might accept two protons to exist as ternary acid H3L2+ with a maximum emission wavelength (lambdamax) at 505 nm. Along with the increase in pH value, fluorescence spectrum of OFL changed, an iso-fluorescence point at 352 nm was found in the fluorescence excitation spectra, and at the same time, isosbestic points were found in its UV absorption spectra. This spectral feature reveals that H3L2+ gradually lost hydrogen ion combined on C-4 keto oxygen. In the range of pH 2.5 to pH 4, OFL exists as H2L+ with lambdamax at 499 nm. When pH>4, the fluorescence emission peak at 499 nm gradually blue-shifted to 455 nm as pH was increasing, and an iso-fluorescence emission point was formed at 484 nm, indicating the dissociation of hydrogen ion of carboxylic acid at C-3. At about pH 7.0, OFL exists in the dipole ion form HL with lambdamax at 455 nm, which is the strongest fluorescence form. When pH>8, lambdamax red-shift from 455 to 475 nm as pH increases, meanwhile, fluorescence intensity decreases, indicating that HL lost hydrogen ion combined on N-4 of piperazinyl group. When pH>10, OFL exists in anion ion form L-, and fluorescence intensity decreases with pH increasing, but lambdamax remains essentially constant, indicating that medium environment influences the fluorescence property of OFL.

The characteristic and mechanism of resonance light scattering (RLS) of Rhodamine B (RhB) were studied. In acidic solutions with pH from -0.38 to 4.10, the intensity of RLS increases with an increase in pH and reaches a maximum at nearly neutral pH. The change in RLS intensity with the wavelength was not accordant with Rayleigh scattering law. The fluorescence excitation and emission spectra of RhB overlap partly, and the RLS peak lies between the fluorescence excitation and emission peaks. In the three-dimensional fluorescence contour spectra of RhB, Rayleigh scattering line intersects fluorescence contour. In light polarization experiment, the polarization of RLS was measured to be P approximately = 0.1. All the above experimental facts reveal that the RLS of RhB is mainly resonance fluorescence. The mechanism of RLS enhancement by the increase in pH is that the formation of fluorescence species occurs in acid-base equilibrium. The RLS peak of RhB appears within the envelope of absorption, and light scattering is affected by light absorption, so, the relationship between RLS intensity and RhB concentration is not strictly linear.

Resonance light scattering (RLS), absorption and fluorescence spectra of aurintricarboxylic acid (ATA) were studied. In solutions with pH 3.7 to pH 11.0, the RLS signal was very weak, but increased sharply with a decrease in pH when pH < 3.7 and reached a maximum at pH 2.7. This enhancement effect of RLS occurred because the negative charged species of ATA were transformed into neutral molecules when pH decreased, and the neutral molecules assembled into supermolecular aggregates. Two peaks at 260 nm and 340 nm, respectively, and a valley at 300 nm, appeared in the RLS spectrum, whereas an absorption peak appeared at 300 nm in the absorption spectrum. This spectral feature reveals that the RLS spectrum was related to the absorption spectrum of ATA. The change in RLS intensity with the wavelength was not accordant with Rayleigh scattering law. The fluorescence excitation and emission spectra of ATA do not overlap, so resonance fluorescence was not involved in the RLS spectrum. Under a given set of experimental conditions, the RLS intensity increased with the increase in ATA concentration, which, however, was not a strictly linear relationship.