Fluorescence and Phosphorescence Spectra

Generally speaking, luminescence spectra (fluorescence and phosphorescence) provide more information about excited states than do absorption spectra. This is because luminescence measurements are much more sensitive than absorption measurements, and the two types of emission can be studied separately due to their widely differing lifetimes. 

FIGURE 3. Fully corrected spectra (fluorescence and phosphorescence) of benzene ip methylcyclo-hexane + 10% chloroform at 77°K, first-... 

Another important stage in systematizing the organic molecules were investigations into the fine structure of their absorption spectra, fluorescence and phosphorescence at 77 and 4.2K44,89 92 tJ3 9[Pg.70]

Luminescence spectra (fluorescence and phosphorescence) measurements provide information about the conflguration of excited states (5i or T ) involved in the reactions. Moreover, the lifetimes of the excited states provide us the information about their origin—(ii Jt ) or (n Ji ). The typical radiative lifetimes of singlet... 

Selectivity The selectivity of molecular fluorescence and phosphorescence is superior to that of absorption spectrophotometry for two reasons first, not every compound that absorbs radiation is fluorescent or phosphorescent, and, second, selectivity between an analyte and an interferant is possible if there is a difference in either their excitation or emission spectra. In molecular luminescence the total emission intensity is a linear sum of that from each fluorescent or phosphorescent species. The analysis of a sample containing n components, therefore, can be accomplished by measuring the total emission intensity at n wavelengths. 

Fluorometry and Phosphorimetry. Modem spectrofluorometers can record both fluorescence and excitation spectra. Excitation is furnished by a broad-band xenon arc lamp foUowed by a grating monochromator. The selected excitation frequency, is focused on the sample the emission is coUected at usuaUy 90° from the probe beam and passed through a second monochromator to a photomultiplier detector. Scan control of both monochromators yields either the fluorescence spectmm, ie, emission intensity as a function of wavelength X for a fixed X, or the excitation spectmm, ie, emission intensity at a fixed X as a function of X. Fluorescence and phosphorescence can be distinguished from the temporal decay of the emission. 

Fluorescence and phosphorescence spectra of poly(propynoic acid)(FPA), polyphenylene (PP), and DPAcN show that the difference of energies between the lower excited singlet and triplet states, as observed in the case of PP (583 nm) and DPAcN (528 nm), is considerably greater than that of poly(propynoic acid) (270—300 nm) which besides transitions may undergo rr - transitions. PCSs showing only... 

UV spectra are, however, very useful for the determination of acid-base and ion pair formation equilibria, and for photochemical investigations (e. g., determination of quantum yield in photolytic dediazoniation, Tsunoda and Yamaoka, 1966 fluorescence and phosphorescence at low temperature, Sukigahara and Kikuchi, 1967a). 

The use of emission (fluorescence and phosphorescence) as welt as absorption spectroscopy. From these spectra the presence of as well as the energy and lifetime of singlet and triplet excited states can often be calculated. 

Burrell and Hurtubise (.32) investigated calibration curves extended well beyond the normal linear range for RTF and RTF of benzoCf)quino-line adsorbed on a silica gel chromatoplate under neutral and acidic conditions. As the benzoCf)quinoline concentration increased, the RTF curves leveled off, whereas the RTF curves passed through a maximum and then decreased. The extended calibration curves along with fluorescence and phosphorescence spectra and phosphorescence lifetimes for benzoCf)quinoline revealed differences in the RTF and RTF phenomena. For example, it was determined that RTF could arise from molecules adsorbed on the surface and in multilayers of molecules, whereas phosphorescence was only generated from molecules adsorbed on the surface of the chromatoplate and not in the multilayers. ... 

Fluorescence and phosphorescence spectra corrected for the instrumental sensitivity were measured with a spectrometer described previously (()). Corrected excitation spectra were obtained with constant excitation intensity controlled by a rhodamine B quantum counter. For phosphorescence polarization measurements the apparatus was set up in an "In Line" arrangement (j ) and equipped with a Glan-Thomson polarizer and a sheet polarizer (analyser) (10). 

The 0-0 transition bands exhibited in the fluorescence and phosphorescence spectra of 4,4 -SD gave the following electronic state energies Sj 33,000 cm-1, Sg 42,000, and ... 

Analyses I.r. spectra were measured as smears on sodium chloride plates or as a solution in carbon tetrachloride using a Perkin-Elmer 567 grating spectrophotometer, while u.v. spectra were measured as a solution in hexane (spectroscopic grade) using a Unicam SP 1700 instrument. Fluorescence and phosphorescence spectra were recorded as described elsewhere (5, 6). 

The fractions from elution chromatography were studied by a number of spectroscopic methods, n.m.r., i.r., u.v., fluorescence and phosphorescence spectroscopy. Equivalent fractions from chromatographic separation of the various oils showed no significant differences in their spectra and it appears that the composition of the fractions was independent of the catalyst concentration used to produce the oil. Though, as previously mentioned the amounts of the various fractions especially the polar fractions differ with the catalyst concentration. G.1.C. analysis of the saturate fractions also indicated no changes with different catalyst concentrations. 

Dependencies of luminescence bands (both fluorescence and phosphorescence), anisotropy of emission, and its lifetime on a frequency of excitation, when fluorescence is excited at the red edge of absorption spectrum. Panel a of Fig. 5 shows the fluorescence spectra at different excitations for the solutes with the 0-0 transitions close to vI vn, and vra frequencies. Spectral location of all shown fluorescence bands is different and stable in time of experiment and during lifetime of fluorescence (panel b)... 

TGA, iodometric, mid-IR, luminescence (fluorescence and phosphorescence) and colour formation (yellowness index according to standard method ASTM 1925) were all employed in a study of aspects of the thermal degradation of EVA copolymers [67], Figure 23 compares a set of spectra from the luminescence analysis reported in this work. In the initial spectra (Figure 23(a)) of the EVA copolymer, two excitation maxima at 237 and 283 nm are observed, which both give rise to one emission spectrum with a maximum at 366 nm weak shoulders... 

The excitation spectrum of a molecule is similar to its absorption spectrum, while the fluorescence and phosphorescence emission occur at longer wavelengths than the absorbed light. The intensity of the emitted light allows quantitative measurement since, for dilute solutions, the emitted intensity is proportional to concentration. The excitation and emission spectra are characteristic of the molecule and allow qualitative measurements to be made. The inherent advantages of the techniques, particularly fluorescence, are ... 

Describe the essential features in the measurement of fluorescence and phosphorescence spectra, including any precautions necessary. 

Fig. 3.1. Perrin-Jablonski diagram and illustration of the relative positions of absorption, fluorescence and phosphorescence spectra.

The aromatic amino acids each have two major absorption bands in the wavelength region between 200 and 300 nm (see reviews by Beaven and Holiday(13) and Wetlaufer(14). The lower energy band occurs near 280 nm for tryptophan, 277 nm for tyrosine, and 258 nm for phenylalanine, and the extinction coefficients at these wavelengths are in the ratio 27 7 l.(14) As a result of the spectral distributions and relative extinction coefficients of the aromatic amino acids, tryptophan generally dominates the absorption, fluorescence, and phosphorescence spectra of proteins that also contain either of the other two aromatic amino acids. 

Figure 1.3. Tyrosine and tyrosinate fluorescence and phosphorescence emission spectra with 280-nm excitation. Curve 1 tyrosine fluorescence in water, pH 7. Curve 2 tyrosinate fluorescence in 0.01 M NaOH. 25 Curve 3 tyrosine phosphorescence in 50% (v/v) ethylene glycol/water/24 Curve 4 tyrosinate phosphorescence in 50% (v/v) ethylene glycol/0.01 A/NAOH/24. ...

Figure 3.2 shows the fluorescence and phosphorescence emission spectrum from tobacco mosaic virus coat protein. These spectra are fairly typical of the tryptophan emission spectra observed from proteins at room temperature. 

Differences between the spectra of fluorescence and phosphorescence are immediately obvious. For all tryptophans in proteins the phosphorescence spectrum, even at room temperature, is structured, while the fluorescence emission is not. (Even at low temperatures the fluorescence emission spectrum is usually not structured. The notable exceptions include a-amylase and aldolase, 26 protease, azurin 27,28 and ribonuclease 7, staphylococcal endonuclease, elastase, tobacco mosaic virus coat protein, and Drosophila alcohol dehydrogenase 12. )... 

THF tetrahydrofuran, NaNap sodium naphthalenide, Na2St disodium stilbene dianion, BP benzoyl peroxide, DME 1,2-dimethoxyethane, TMBD tetramethylbenzidine dication diperchlorate, WBP Wurster s blue perchlorate, LP lauroyl peroxide, FLSPEC fluorescence spectrum obtained, FPSPEC fluorescence and phosphorescence spectrum obtained, FXSPEC fluorescence and eximer spectra obtained, DPAC12 9,10-dichlorO 9,10-dihydro-9,10-diphenylanthracene. 

Fluorescence and Phosphorescence. Essentially the same effects that influence absorption of radiation are found to perturb the emission processes. Fluorescence spectra are red- or blue-shifted along with the corresponding band of the absorption spectrum. 

The fluorescence and phosphorescence spectra of a complex molecule are generally discussed by reference to an energy level diagram such as that shown in Figure 1. Absorption of light raises the molecule from the ground state to one of the upper electronically excited singlet states. At... 

The influence of the presence of nitrogen atoms on the fluorescence and phosphorescence spectral maxima of aromatic molecules has been discussed, including comparison of the spectra of phenanthrene and 1,10-phenanthroline, the spectral maxima of which are similar.77... 

In the vapor phase, the emission from excited acetone has been shown to be a mixture of fluorescence and phosphorescence [254]. The estimated lifetime of the excited singlet state is 10 ns, a figure commonly accepted as a reasonable approximation to the excited singlet to triplet (St - TJ transition time in aliphatic ketones. The overlap of the fluorescence and phosphorescence spectra reflects the fact that the energetic separation between the lowest n, n single and triplet states is small, at least in comparison with the S2 — Tt splitting between lowest excited n, n singlet and triplet states. 

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