ISSN 0030-400X, Optics and Spectroscopy, 2008, Vol. 104, No. 5, pp. 717723. Pleiades Publishing, Ltd., 2008.

. Original Russian Text V.I. Tomin, 2008, published in Optika i Spektroskopiya, 2008, Vol. 104, No. 5, pp. 793800.

CONDENSED-MATTER SPECTROSCOPY

Dynamic Quenching of the Dual Fluorescence of Molecules

V. I. Tomin
Institute of Physics, Pomeranian University, S l upsk, 76-200 Poland e-mail: tomin@apsl.edu.pl
Received July 18, 2007

AbstractMathematical relations describing the properties of spontaneous steady-state dual uorescence under conditions of dynamic quenching of excited states by foreign impurities are derived. It is shown that, in the case of a kinetic character of the reaction, the initial form of the dye and its photoproduct are quenched, the intensity ratio of the uorescence bands of the initial form and the product linearly increasing with the quencher concentration. Analysis performed is applicable to a wide range of photoreactions accompanied by the dual uorescence (charge transfer, proton transfer, complexation, etc.). The properties of the uorescence, absorption, and dual uorescence excitation for 3-hydroxyavone in acetonitrile under conditions of dynamic quenching by the TEMPO spin quencher with a concentration below 1.25 102 M are studied. 3-Hydroxyavone is characterized by the excited-state intramolecular proton transfer and by the uorescence spectrum consisting of two well-spaced bands. The observed dependences of the intensity of both uorescence bands on the quencher concentration correspond to the theoretical conclusions. The SternVolmer constants calculated from the experimental data on the assumption of diffusion quenching of the excited states are 858 and 1141 M1 for the normal and tautomeric uorescence bands, respectively. The experimental results reveal the kinetic character of the excited-state proton transfer in 3-hydroxyavone in acetonitrile. PACS numbers: 33.50.Dq DOI: 10.1134/S0030400X08050123

INTRODUCTION In recent decades, various photoreactions in excited states of organic compounds and their complexes, such as the charge transfer (CT), the proton transfer (PT), the complexation, and some others, have been very intensively studied owing to the fact that they are very common among primary photoreactions. The understanding of such reactions is of fundamental importance [1, 2] because they play a key role in the main photosynthesis reactions of plants and in the functioning of various biological organisms; currently, they are also extensively used in molecular electronics and biotechnologies. The excited state photoreactions in individual molecules strongly change their physicochemical properties. Rather frequently, both the initial molecules and the reaction products exhibit pronounced dual uorescence with well-spaced spectral bands. Investigation of its properties provides rich information about accompanying processes, including photoreactions, and, hence, is very important. All this stimulates a permanent interest to the simplest photoreactions in polyatomic molecules and their active investigation. Along with investigation of the fundamental properties of these photoreactions, signicant efforts are focused on the creation of new methods for studying physicochemical and biological systems, as well as on the development of principles of investigation of such systems by molecular probes using the CT, PT, and some other reactions.

Currently, it is believed that some specially synthesized derivatives of the 3-hydroxyavone (3-HF) family completely satisfy the requirements imposed on multiparametric probes [3, 4]. To ensure a highly sensitive response, the probe molecule should have the possibility of existing in different ground and excited states depending on the conditions. Transitions between these states form absorption and emission bands, which can with different sensitivity respond to perturbations from the microenvironment. It is these properties that are inherent in molecules of some 3-HF derivatives. The dual uorescence of 3-HF was observed and described in [5, 6]. To date, the 3-HF molecule has been the most actively studied in order to understand the dynamics and mechanism of intramolecular PT. Upon excitation of 3-HF derivatives into the absorption band maximum [79], one usually easily observes two uorescence bands due to the excited-state internal proton transfer (ESIPT). Figure 1 shows the energy level diagram that describes the PT reaction and reects the existence of four energy levels: the ground N and excited N* levels of the normal form and the excited T* and ground T levels of the PT or tautomeric form. The structural formulas of the parent 3-HF molecule and of the tautomer are shown at the bottom of Fig. 1. According to the classical scheme proposed in [5, 6], after absorption of light, the 3-HF molecule passes from the ground state N to the excited level N*, which is responsible for the short-wavelength (blue) uores-

717

718 N* T*

TOMIN

ex

II

T N
O

OH

OH

Fig. 1. Energy level diagram of 3-HF (N and N*) and its excited-state PT tautomer (T and T *).

cence band (the N* N transition with the frequency I). From this state, a fast adiabatic transition occurs to the excited state T * of the PT form. The PT photoreaction occurs for about 50100 fs [10, 11]. Hence, the PT rate constant can be estimated to be of the order of 1013 s1. The second luminescence band corresponds to the transition from the excited PT state T * to the ground state T (Fig. 1). This is a green uorescence band with the frequency II, which is comparable in intensity with the short-wavelength band, but is considerably (by ~130 nm) shifted to longer wavelengths so that the bands overlap only slightly. A very important feature of 3-HF derivatives is the possibility of self-calibration of their luminescence signal because the effect of the environment causes a change in the intensity ratio of the two bands IN /IT. This ratio is determined by intramolecular processes and intermolecular interactions with the environment and does not depend on the instrumental factors; since these probes are based on the ratio IN /IT, they are sometimes called ratiometric. The possibility of self-calibration considerably simplies absolute measurements using these probes. The second important property of such probes is their ability to respond to various intermolecular interactions by variations in the main features of the two uorescence bands an in the mutual positions of the energy levels of the N and T forms. These interactions can change the relative positions of the levels and the energy gaps between them so strongly that the type of PT reaction radically changes. As is known, depending on these parameters, the PT reactions can be divided into two types. The rst type implies kinetic PT reactions, which are characterized by a high rate constant and a weak contribution of the N band to the steady-state spectra in comparison with the contribu-

tion of the T band. In this case, the two uorescence bands decay by different laws with one common fast component corresponding to the PT. The second type is the so-called thermodynamic reactions, which are characterized by a strong interaction between the excited states of the N and T forms and, hence, by identical decay laws. The two types are usually identied using uorescence measurements with a high time resolution in the pico- and femtosecond ranges. Rather often, the intra- and intermolecular interactions can considerably change the decay laws and decrease the quantum yields of luminescence of atoms and molecules, thus leading to the luminescence quenching. There are different types of quenching, such as temperature quenching, concentration quenching, and quenching by foreign impurities, in particular, by oxygen. As a rule, all the quenching types can be simultaneously observed in quantum systems. The knowledge of the quenching mechanisms allows one to use them in investigation of photoreactions in excited states of molecules. The most important for this purpose is the so-called dynamic quenching, when the quenching occurs due to the interaction of excited molecules with specially chosen impurities. In [12], the uorescence, absorption, and excitation spectra of 3-HF in acetonitrile with addition of a classical quencher, potassium iodide KJ, with concentrations of up to 4 102 M were studied in the UV and visible regions. The intensities and quantum yields of both bands showed rather complex dependences on the quencher concentration, the stronger quenching being observed for the long-wavelength uorescence band, which belongs to the tautomeric form. In this study, based on the balance equations, we derive expressions for the populations of excited states of a molecular probe with the excited-state PT. The 3HF uorescence quenching by the 2,2,6,6-tetramethyl1-piperidinylooxy (TEMPO) spin quencher is studied experimentally and the dependences of the quenching efciency on the quencher concentration are determined. It is found that the intensities and quantum yields of the two bands almost linearly decrease with the quencher concentration and that the long-wavelength uorescence band corresponding to the tautomeric form of 3-HF is quenched much more efciently. The behavior of quenching of the two forms allows us to conclude that the PT reaction in the 3-HF solution in acetonitrile has a kinetic character. THEORY We will consider a molecule in which an excitedstate photoreaction occurs according to the four-level scheme shown in Fig. 1. The conclusions made in this section will be applicable only to this reaction. However, for deniteness, we will assume that this reaction is the PT because, below, we will compare our theoretical results with the experimental data obtained for
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DYNAMIC QUENCHING OF THE DUAL FLUORESCENCE OF MOLECULES

719

solutions of 3-HF, which is a well-studied classical molecular object with the excited-state PT occurring as is shown in Fig. 1. The time dependences of the number of molecules in the excited N* and T * states can be obtained using the following kinetic equations taking into account the rate constants (or probabilities) of all the transitions for PT illustrated in Fig. 1: dN */ dt =
(N) [kR

hence, about the corresponding intensities of the N and T fluorescence bands. Consider the simple but rather frequent case when the direct PT rate is considerably higher than the rate of all the other processes of deactivation of the N* state, i.e., k + k R + k nR + k q Q .
(N) (N)

(9)

(N) k nR

+ k+ ] N *

(1) (2)

+ k T * + k a u ex N 0 , dT */ dt =
(T ) [kR

For this situation, Eqs. (7) and (8) can be presented in the form N * = ( k a u ex N 0 / k + ) { 1 + k / [ k R + k nR + k q Q ] } , (10) T * = k a u ex N 0 / [ k R + k nR + k q Q ] .
(T ) (T ) (T ) (T )

(T ) k nR

+ k ] T * + k + N *.

In Eq. (1), the product kauexN0 describes the number of absorption transitions to the excited state of the N form (ka is the absorption coefcient, uex is the volume density of excitation light, and N0 is the concentration of molecules in the ground state of the initial form), k R
N (T ) N (T )

(11)

and k nR are the rate constants of, respectively, radiative and nonradiative transitions from the excited states of the N and T forms, and k+ and k are the rate constants of the direct and reverse PT reactions. Let us introduce in our solution a quencher with a concentration Q and let this quencher cause dynamic quenching of both forms due to diffusion collisions. We will consider the steady-state uorescence of this solution. In this case, the populations of excited states N* and T * can be found using the balance equations { [ k R + k nR + k + ] + k Q Q } N * = k T * + k a u ex N 0 , (3) k + N * = { [ k R + k nR + k ] + k Q Q } T *.
(T ) (T ) (N) (N)

As is seen from these relations, the uorescence of the N and T bands may be quenched with increasing quencher concentration Q. These relations can be easier and simpler analyzed in limiting cases. Let us consider two characteristic cases. In the rst case, the reverse PT constant is close to the sum of the constants of the T * state deactivation k k R + k nR + k q Q .
(T ) (T )

(12)

Take also into account that, in ordinary experiments, the rates of the diffusion quenching [13, 14] and of the excited state deactivation are of the same order, k q Q k R + k nR .
(T ) (T )

(13)

For simplicity, we also assume that the deactivation rates for the N and T states do not differ strongly, i.e., k R + k nR k R + k nR . Taking into account the latter condition, Eq. (9), and assumptions (12) and (13), we can clearly see that the direct PT rate considerably exceeds the rate of the reverse reaction, k+ k. This equation corresponds to the kinetic model of PT. Then, taking into account the latter inequality and (12), we nd from (10) and (11) that the tautomeric form should be quenched more strongly than the normal form. Another interesting case is observed for a rather high constant of the reverse PT transition, k ~ k+. In this case, as follows from (10), (11), and assumption (9), the N* and T * populations identically respond to the presence of a quencher, i.e., the decay law is the same for both forms. Hence, the PT has a thermodynamic character. The most interesting characteristic parameter of PT probes is the relative intensity of the N and T uorescence forms. This parameter can be easily obtained
(T ) (T ) (N) (N)

(4)

Denote the terms in the braces in Eq. (3) as {}1 and the terms in the braces in Eq. (4) as {}2. Then, Eqs. (3) and (4) can be written in a more compact form { } 1 N * = k T * + k a u ex N 0 , k + N * = { } 2 T *. (5) (6)

The obtained system of linear algebraic equations with respect to unknown populations N* and T * has the solutions { }2 k a u ex N 0 - k u N , (7) N * = ------------------------------------ = ---------------------------------{ } 1 { } 2 k k + a ex 0 { }1 k k + / { }2 { }2 k+ - ----------------------------------k u N T * = -------{ } 2 { } 1 { } 2 k k + a ex 0 k + k a u ex N 0 -. = ---------------------------------{ }1 { }2 k k +

(8)

From these equations, taking into account the rates of all reactions and transitions between the levels of the scheme shown in Fig. 1, we can obtain all the necessary information about the N* and T * excited states and,
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720

TOMIN

H3C H3C

N O

CH3 CH3

Fig. 2. Structural formula of the TEMPO quencher.

they overlapped with the uorescence bands. The measurements were performed at room temperature (295 K). In all the cases, the cell with the sample was thermostatically controlled. The uorescence was recorded using a standard scheme at an angle of 90 to the excitation beam. All the results were reproduced in several experimental series. EXPERIMENTAL RESULTS The absorption spectra of 3-HF in acetonitrile with different content of the TEMPO quencher are shown in Fig. 3. As is seen, TEMPO has its own absorption with a wide band, characteristic of organic substances, at 460 nm with the absorption coefcient of 0.16 cm1 at a concentration of 2 102 M. The second absorption band arises near 320 nm and has the maximum at 240 nm with an intensity of an order of magnitude higher than the intensity of the long-wavelength band. The inset in Fig. 3a shows the dependences of the absorption coefcients at the wavelengths of 340 and 460 nm on the TEMPO concentration. It is seen that TEMPO almost does not change the dye absorption in the maximum of its main band near 340 nm. At the same time, the absorption in the maximum of the longwavelength band of TEMPO linearly increases with its concentration. We should also point out a noticeable but small enhancement of the quencher absorption in the region of the maximum of the main absorption band of 3-HF, which should slightly affect the total absorption spectrum of the solution. The uorescence spectra of 3-HF in acetonitrile at different quencher concentrations are shown in Fig. 4. As is seen, the standard excitation near 340 nm is accompanied by the appearance of the well-known spectrum with the blue and green bands near 395 and 525 nm, respectively. The rst peak belongs to the normal form of the 3-HF molecule and the second peak corresponds to the tautomeric form formed due to the PT. The second peak has a signicantly higher intensity than the rst peak. The quencher causes a strong quenching of both bands, especially of the green one. The dependences of the uorescence intensity in the maxima of these bands on the TEMPO concentration are presented in Fig. 5. Figure 5a shows a continuous decrease in the intensity of the N form with increasing concentration C[TEMPO], excluding the concentration range from 2 103 to 5 5 103 M; the most pronounced decrease in the uorescence intensity is observed in the concentration range 0103 M. The decrease in the intensity of the green band (Fig. 5b) is stronger in comparison with the violet band in the whole concentration range. Hence, the tautomeric form is more sensitive to the quencher concentration. The strongest effect of quencher is observed at the concentrations C[TEMPO] ranging from 0 to 103 M and 1021.25 102 M. It should be noted that the proOPTICS AND SPECTROSCOPY Vol. 104 No. 5 2008

from the population ratio N*/T *. At rst, write the population ratio N*/T * using relations (10) and (11), N */ T * = { [ k R + k nR + k ] + k q Q } / k + .
(T ) (T )

(14)

Taking this into account, we can write the intensity ratio of the N and T forms as I N / I T = ( N */ T * ) ( k R / k R ) ( I / II ) = ( k R I / k + k R II ) { [ k R + k nR + k ] + k q Q } .
(N) (T ) (T ) (T ) (N) (T )

(15)

The term at the braces can be taken to be constant for a specic solution. As is seen, the ratio IN /IT linearly changes with the quencher concentration Q. Obviously, the role of the quencher, which is determined by the product kqQ, will be noticeable if this product exceeds or is comparable with the sum of the three terms in the square brackets of expression (15). Naturally, this requirement is fullled only for rather low reverse reaction rates k, for example, under condition (12). From (15), it is also seen that, at rather high rates k (k ~ k+), the role of the quenching factor is negligibly small and the intensity ratio IN /IT does not depend on the quencher concentration, although each uorescence band is quenched. Such a behavior characterizes the thermodynamic type of the PT reaction in the solution studied. Relations (10), (11), and (15), obtained in this section, were experimentally tested in investigations of the quenching of the dual uorescence of 3-HF in acetonitrile when the excited state of this probe was decayed due to the TEMPO spin quencher (we measured the uorescence intensities, which are proportional to the corresponding populations determined by (10) and (11)). The structural formula of this organic quencher is shown in Fig. 2. EXPERIMENTAL We used solutions of 3-HF (Indole Chemical C., additionally puried by recrystallization) in acetonitrile (Scharlau, pure grade, for UVVIS spectroscopy) with a concentration of 105 M. As a quencher, we used TEMPO, analytical grade, Aldrich. The excitation and uorescence spectra were recorded using a Hitachi F-2500 spectrouorimeter and the absorption spectra were measured with a Hitachi U2810 spectrophotometer. The pure solvent had virtually no impurities luminescing under excitation in the entire wavelength region from 350 to 670 nm. The Raman spectra were subtracted from the total spectra when

DYNAMIC QUENCHING OF THE DUAL FLUORESCENCE OF MOLECULES ka, cm1 () ka, cm1 0.25 0.20 0.15 2 2 0.10 7 6 5 4 3 1 0.05 0 0.3 0.9 C [TEMPO] 102, 7 6 5 4 3 2 1 2 1

721

1 0 220 340 (b) 460 7

580

0.15

6 7 5 4 1 3

0.10

0.05 2 0 250 1 370 490 , nm

Fig. 3. (a) Absorption spectra (16) of 3-HF in acetonitrile with C[TEMPO] = (1) 0, (2) 103, (3) 5 103, (4) 7.5 103, (5) 102, and (6) 1.25 102 M and (7) of the quencher in acetonitrile. (b) The same on an enlarged scale for the wavelength region from 250 to 640 nm. The inset in Fig. 3a shows dependences of the solution absorption at the wavelengths (1) 340 and (2) 460 nm on C[TEMPO]. C[3-HF] = 105 M.

les and positions of the N and T uorescence bands do not noticeably change in the quenching process for any used concentrations of TEMPO. The dependence of the intensity ratio of the maxima of the N and T uorescence bands IN /IT on the TEMPO concentration is shown in Fig. 6. This dependence is obtained using the data presented in Fig. 5. As is seen, the ratio IN /IT increases with the quencher concentration according to an approximately linear law.
OPTICS AND SPECTROSCOPY Vol. 104 No. 5 2008

DISCUSSION It is well known that some impurities can attenuate spontaneous emission of solutions; this process is called the uorescence quenching. The quenching occurs as a result of intermolecular interactions and can be of the rst or second kind [2, 13, 14]. The quenching of the rst kind (or the static quenching) is a process occurring due to the effect of quencher on the molecules in the ground state. In fact, in this case, we deal with complexation involving the ground state of the

722 Ifl, arb. units 1000

Ifl, arb. units 1

TOMIN

1 2 3 4 5

800

10

600

0 350

400

, nm

400

200

0 350

450

550

650

, nm

Fig. 4. Fluorescence spectra of 3-HF in acetonitrile with the quencher concentrations C[TEMPO] = (1) 0, (2) 103, (3) 5 103, (4) 7.5 103, (5) 102, and (6) 1.25 102 M. The inset shows the short-wavelength uorescence band on an enlarged scale. The excitation wavelength is 340 nm. C[3-HF] = 105 M.

dye. The principal difference between the two kinds of quenching is that the quenching of the second kind (dynamic) occurs when the foreign material interacts with the excited states of the uorophore. In this case, the uorescence yield is decreased due to the nonradiative deactivation of excited molecules, which can occur via different channels, namely, by the energy transfer from the excited uorophore molecules either to unexIfl, arb. units 30 25 20 1000 900 800 700 0 0.3 0.5 0.8 0.11 0.14 C [TEMPO] 102, (b)

cited molecules or to quencher particles, where the energy transforms to thermal vibrations. In the case of the dynamic quenching, which is a pure physical process, the absorption and uorescence spectra do not change, while the uorescence quantum yield decreases in parallel with a shortening of the excited state lifetime [13, 14]. The absorption spectra shown in Fig. 2 consist of two intense electronic bands, which can be assigned to the S0 S1 and S0 S2 singlet bands of 3-HF. This is proved, for example, by the results of semi-empirical quantum-mechanical calculations of the 3-HF molecule [15]. The behavior of the absorption in the maximum of the normal band (curve 1 in the inset in Fig. 3) testies that the absorption at the excitation wavelength changes only insignicantly with the quencher concentration. Hence, the addition of TEMPO does not affect the absorption of 3-HF, in contrast to the ionic quencher KI [10], whose effect on the 3-HF absorption is unusualthe absorption spectrum prole remains the same while the absorption increases at all the wavelengths, although the quencher itself has no absorption in this region. Next, let us consider the behavior of the uorescence spectra in solutions with a quencher. Since the absorption almost does not change with addition of the quencher, the dependences in Fig. 5 reect the behavior of the quantum yield of the corresponding uorescence bands and, hence, the linear approximation of these curves in the assumption of the diffusion mechanism
OPTICS AND SPECTROSCOPY Vol. 104 No. 5 2008

()

Fig. 5. Dependences of the uorescence intensity I in the maxima of the (a) normal (395 nm) and (b) tautomeric (525 nm) forms of 3-HF in acetonitrile on the TEMPO concentration. The uorescence excitation wavelength is 340 nm. C[3-HF] = 105 M.

DYNAMIC QUENCHING OF THE DUAL FLUORESCENCE OF MOLECULES Ifl, arb. units 0.03

723

0.02 0

0.2

0.5

0.8 0.11 0.14 C [TEMPO] 102,

Fig. 6. Intensity ratio of the uorescence of the normal and tautomeric bands IN /IT as a function of the quencher concentration. The intensities were measured in the maxima of the corresponding bands. The dependence is plotted based on the data given in Fig. 5.

(2) The properties of the uorescence, absorption, and dual uorescence excitation for 3-hydroxyavone in acetonitrile under conditions of dynamic quenching by the TEMPO spin quencher are studied. The intensities of the two uorescence bands and their quantum yields depend on the quencher concentration in good agreement with the theoretical results. The Stern Volmer constants calculated based on the experimental data are 858 and 1141 1 for the normal and tautomeric uorescence bands, respectively. The experimental results allow us to conclude that the excited-state proton transfer in 3-HF molecules in acetonitrile has a kinetic character. ACKNOWLEDGMENTS I acknowledge nancial support from the Pomeranian University (S l upsk) (project no. BW 6/20/06). I thank G. Smolyarchik for spectral measurements. REFERENCES
1. Molecular Interactions, Ed. by H. Ratajczak and W. J. Ortwille-Thomas (Wiley, New York, 1983), Vol. 2. 2. J. Lakowicz, Principles of Fluorescent Spectroscopy (Plenum, New York, 1984). 3. A. P. Demchenko, Anal. Biochem. 343, 1 (2005). 4. A. P. Demchenko, A. S. Klymchenko, V. G. Pivovarenko, et al., J. Fluoresc. 13, 291 (2003). 5. P. K. Sengupta and M. Kasha, Chem. Phys. Lett. 68, 382 (1979). 6. M. Kasha, J. Chem. Soc., Faraday Trans. 282, 2379 (1986). 7. A. S. Klymchenko and A. Demchenko, Phys. Chem. Chem. Phys 5, 461 (2003). 8. A. S. Klymchenko and A. P. Demchenko, New J. Chem. 6, 687 (2004). 9. A. P. Demchenko, Fundamental Photoprocesses and Inhomogeneous Broadening of Electronic Spectra of Organic Molecules, Ed. by V. I. Tomin (Wyd. OAP, 2006), p. 79. 10. S. Ameer-Beg, S. M. Ormson, R. G. Brown, et al., J. Phys. Chem. A 105, 3709 (2001). 11. A. N. Bader, G. Ariese, and C. Gooijer, J. Phys. Chem. A 106, 2844 (2002). 12. V. I. Tomin and G. Smolyarchik, Opt. Spektrosk. 104 (6), (2007). 13. A. N. Terenin, Photonics of Dye Molecules and Related Organic Compounds (Nauka, Leningrad, 1967) [in Russian]. 14. B. I. Stepanov, Luminescence of Complex Molecules (Akad. Nauk BSSR, Minsk, 1956) [in Russian]. 15. N. Chattopadhyay, M. Barroso, C. Sepra, et al., Chem. Phys. Lett. 387, 258 (2004).

can be used to estimate the quenching constants kq by the SternVolmer formula B0 / B = 1 + k q Q , where B and B0 are the quantum yields of the uorophore with a quencher and without it and Q is the quencher concentration. The thus calculated Stern Volmer constants have typical values and are equal to 858 and 1140 1. These values agree with the data that are usually obtained on the assumption of the diffusion collision mechanism of quenching of excited states [2, 13]. Consider the dependence of the intensity ratio IN /IT on the quencher concentration (Fig. 6). It is seen that this ratio is higher than for the pure solution and monotonically increases with the TEMPO concentration. In the whole concentration range, this increase is not too signicant, from 0.024 to 0.03. Therefore, we can conclude that the uorescence intensity ratio depends on the presence of the quencher in the solution and is noticeably sensitive to a change in its concentration. Thus, our experimental results prove that the mechanism of PT in the solution of 3-HF in pure acetonitrile and with addition of the spin quencher corresponds to the kinetic model. This means that the reverse PT constant k is rather small in comparison with the direct transition probability k+, i.e., the energy level T * lies considerably lower than the T * level. CONCLUSIONS (1) Mathematical relations describing the properties of the steady-state spontaneous emission of systems with dual uorescence under the conditions of dynamic quenching of their excited states by impurities are derived. It is shown that the neutral form of molecules and their photoreaction product undergo quenching, whose intensity is higher for the reaction product. The intensity ratio of the uorescence bands of these forms is directly proportional to the quencher concentration.

Translated by M. Basieva

OPTICS AND SPECTROSCOPY

Vol. 104

No. 5

2008