The study investigates the excited state proton-transfer spectroscopy of 3-hydroxyflavone and quercetin, revealing that their yellow-green luminescence at room temperature is due to a tautomer produced by this transfer across a hydrogen-bonding barrier. The research highlights the significant viscosity dependence of this tautomerization process and its implications for fluorescence behavior. Additionally, deuteration effects were observed, indicating that external hydrogen bonding can inhibit tautomerization, further complicating the understanding of the luminescence mechanisms in these molecules.
The study investigates the excited state proton-transfer spectroscopy of 3-hydroxyflavone and quercetin, revealing that their yellow-green luminescence at room temperature is due to a tautomer produced by this transfer across a hydrogen-bonding barrier. The research highlights the significant viscosity dependence of this tautomerization process and its implications for fluorescence behavior. Additionally, deuteration effects were observed, indicating that external hydrogen bonding can inhibit tautomerization, further complicating the understanding of the luminescence mechanisms in these molecules.
The study investigates the excited state proton-transfer spectroscopy of 3-hydroxyflavone and quercetin, revealing that their yellow-green luminescence at room temperature is due to a tautomer produced by this transfer across a hydrogen-bonding barrier. The research highlights the significant viscosity dependence of this tautomerization process and its implications for fluorescence behavior. Additionally, deuteration effects were observed, indicating that external hydrogen bonding can inhibit tautomerization, further complicating the understanding of the luminescence mechanisms in these molecules.
The study investigates the excited state proton-transfer spectroscopy of 3-hydroxyflavone and quercetin, revealing that their yellow-green luminescence at room temperature is due to a tautomer produced by this transfer across a hydrogen-bonding barrier. The research highlights the significant viscosity dependence of this tautomerization process and its implications for fluorescence behavior. Additionally, deuteration effects were observed, indicating that external hydrogen bonding can inhibit tautomerization, further complicating the understanding of the luminescence mechanisms in these molecules.
Department of Chemistry a& Institute of Molecub BiopIt>sics. Flonila State Uniwrsitv, Talhhmee, FIonda 32306, USA
Rcceited 4 September 1979
The yellow-green luminescence of 3-hydroKyfkwone zmd quercerin ztrroom tempemture in solution arises from a tautomcr of the mol=cuies produced by excited state proton-tmnsfcr zwzross ;1barrier in the double-minimum hydrogen- bonding potenti& At 77 K in 2-methylbutane rigid mat& 3.normal W-violet fluorescence is observed in correspondence with the W absorption Ewzitition spectra and deutemtion effects confum the mechanism.
The molecules 3-hydroxyflavone and quercetin ex-
hibit a brilliant yellow to yel!ow-green apparent fluo- rescence in fhrid soiutions at rocm temperature, yet have their fii singIet--singIet absorption band peak at about 350 nm, \Ve have discovered that the yellow region Iuminescence in these molecules is from a tautomer of the parent molecule, produced by ex- quercetii 3-hydroxyflovone cited state proton transfer across a barrier in the dou- ble-minimum hydrogen-bonding potential_ Our curiosity was aroused by the standard diagnos- study is 3-hydroxyflavone, as the skeletal precursor tic test for the plant fkwone quercetin in thin-layer- of the flavonols- chromatography. We have been employing Eastman The W absorption spectrum for this molecule is Kodak TLC plates of cellulose in a project of flavone given in fig_ I, and shows a characteristic vibrationai en- separation and identification of flower petal methanol velope profile with a first peak at 352 nm, with the on- extracts from hybrids of genus fZemerocallis. The ob- set of singlet-singlet absorptionat 370 nm. (In querce- servation of yelIow to yellow-green Iuminescence tin the absorption peak is at 380 n-with onset at 420 spots is taken as the preliminary diagnosis for the nm_) The room temperature luminescence spectrum flavonols, e-g_, kaempferoi, quercetin, myricetin- This of 3-hydroxyflavone is given in fig_ 2b with a peak at yellow luminescence could not be the normal singlet- 520 nm in the green region. Following the observa- singIet fluorescence of thz corresponding initial mole- rions of Song et al_ [I] on the unusual viscosity de- c&s, since these are W absorbers and should have pcndence of anthocyanidin pigments, we sought the their normal fluorescence in the near W or the violet anaIogous behavior in 3-hydroxyflavone. \Ve pictured region_ (fig- 3) the imernally hydrogen-bonded compiex of 3- The molecule we chose for a detaiIed spectroscopic hydroxyffavone as the initial moIecular species, which could then tautomerize in the excited state, possibly going over to the benzopyrylium eiectromer of the * This work w supported by s contact between the Division of Biomedi& snd Enviromnenti Resezuch, Depment of tautomer- This could account for the large spectral Energy, U.S., and the Rori& Smte University_ shift, as the green to yellow luminescence approaches
382 Volume 68, number 2,3 CHEMICAL PHYSICS LETTERS 15 December 1979
300 -350 4 Fig. 2. Luminescence spectrum of 3-hydroxyflavone, 2.0 X ‘hWELENGTti. nm 10m5 hl in Zmethylbutane solution_ (a) Violet fluorescence Fis 1. Uitmviolet absorption spectrum of Shydro*yflavone of normal molecule at 77 K in rigid glass matri.. 1%) Green in 7-methylbutane solution (1 cm cell) at room temperature fluorescence of tautomer ar room temperature (297 K). (297 fi). (Cat-y 15 spectrophotometer.) (Varian SF 330 spectrofluorimeterr) (The hump at 170 nm in a is probably the phosphorescence of the normal molecule
Fig 3. Excited state proton transfer model for 3-hydroxyflavone (icreasin, 0 tautomerization barrier heights 1,2,3 corresponding to increasing viscosities [ 2])_ - 383 Vohame 65. number 2-3 CHEMICAL PHYSICS LElTERS 15 December 1979
the spectral region of the anthocyanidins which have
the benzopyrylium electronic structure_ We perceived that the viscous-barrier spectrosmpic- cage model of Dellinger and Kasha [2] could apply, and that at increasing viscosities the barrier height (1, 2-3 of fig_ 3) for tautomerization is increased_ Song et al_ [I] pictured the origin of the viscosity barrier to be the torsional motion of the phenyl group about its junction axis to the main heteroatom ring system- Accepting this phenyI torsion as being suftkient cause for a viscosity dependence, we picture that in the sin- glet excited state the phenyl ring must become co- phtnar with the -y-pyrone ring of 3hydroxyflavone in order that the fuii b&city of the carbons1 oxygen can develop, thereby permitting the proton transfer- If, however, torsion of the phenyl ring is prevented so that the out-of-plane conformation is frozen in before excitation, then the tautomerization should be inhib- Fig. 4. Deutemtion effects on luminescencespectraof 3- ited_ We then did observe that in Zmethykutane gbiss hydroxyflavone, 2 X IO-’ bl at room temperature (397 I(). at 7-7 K the normal (violet-blue) fluorescence of 3- (a) Methanol solvent, sI10whu~rautomerization competition hydroxyflavone could be obtained (fig- 2a), corre- xiith external H-bonding_ (b) Methanol-D sohent. showing tunneIing barrier to tautomerization in competition with ex- sponding exactly to the absorption spectrum (fig- 1) ternal D-bondins (Baird Atomic SFR-LOO spectrofluorimeter.) in the normal vibration envelope inversion relation- ship_ A very smaII residual green tautomer emission could still be seen, as well as evidence of phosphores- emission of 3-hydroxyffavone in methanol and meth- cence peaks at 475 nm_ Thus, shift from the normal anol-D as solvents at room temperature_ The hydroxy fluorescence to proton-transfer tauto_mer fluorescence group is expected to be fully deuterated. In these hy- is found to be extraordinarily viscosity dependent- drogen-bonding soIvents two emissions are ahvays ob- Froiov et al. [3] presented some related preliminary served, in agreement with the report of Frolov et al. observations on the luminescence behavior of 3- The interpretation now is more complex. Firstly, we hydroxyflavone and related mole&es at 77 K in see that external hydrogen-bonding to the solvent ethano1 glass_ They observed two Iuminescences of (fig. 4a) can prevenr tautometization; this point was equivalent intensity for 3-hydroxyflavone in ethanol missed by Frolov et al., and their ad hoc intemal- glass and believed them to be emissions from the in- external H-bond assumption provides no explanation ternally and externally hydrogen-bonded sohtte. of the large spectral shift_ Secondly, we see that deu- To further characterize the tautomerization we territionaIso inhiiits the excited state proton transfer studied the excitation spectra for the two Iumines- as expected Fora turaeling process (fig. 4b). We are cence spectral regions of 3-hydroxyflavone and deu- continuing these studies and are endeavoring to sepa- teration effects on the luminescence_ The excitation rate the effects described by studying 3deuteroxy- spectrum for the green region luminescence and the flavone in hydrocarbon solvent. violet region hrminescence, both for hydrocarbon The case of excited state proton-transfer in 3- @ass solution at 77 K (fig. 2a), were found to be iden- hydroxyflavone is in striking analogy to the similar tieai,indicating that they arise from the same primary phenomenon in 7-azaindole [4,5] dimer and the etha- molecular species_ In EPA glass and 2-methylbutane nol sohfate of %azaindole_ In 7-azaindoIe the cbge glass, well-resoIved excitation spectra are observed from a violet fluorescence of the normal molecule to corresponding exactly with the 3-hydroxyflavone ab- a green fluorescence of the tautomerized molecule was sorption curve. Deuteration effects have also proved interpreted [4] in terms of an analogous double- to be strongly evidenced_ Fig. 4 compares the relative minimum potential model- in the latter case, however,.
384 VoIume 68, number 2,3 CHEMICAL PHYSICS LETTERS 15 December 1979
the viscosity effect was observed on the barrier height, [2] B. DeUinger and hI. Kasha, Chem. Phys. Letters 36 but no internal motion was diagnosed which could (1975) 410; 38 (1976) 9. [3] Yu.L. FroIov, YuM. Sapozhnikov, S.S. Barer, N.N. offer an origin for such a variation of the tautomeriza- Pagodaeva and N_k Tyukavkina, Izv. Akad. Nauk USSR tion barrier_ In the present case of 3-hydroxyflavone Ser. Khirn. 10 (1974) 2364. the very great sensitivity to viscosity of excited state [4] CA Taylor, MI. Ashraf El-Bayoumi and hi. Kasha, Proc, proton-transfer suggests the possibility of a presszlre Natl. Acad. Sci US 63 (1969) 253. modulation of the violet to green luminescence con- [S] M. Ashwf El-Bayoumi, J. Phyr Chem. 80 (1976) 2259. version. [61 V.P. GeorgieM5i and AL Rybachenko, Zh. Pni Spektroskopiya 22 (1975) 761
References
[ 1] P.-S_ Song, T.A. Moore and hi. Sun. in: The chemistry of plant pigments, advances in food research, SuppL 3, ed. CO_ Chicbester (Academic Press, XeewYork. 1972) pp_ 33-74 (ccf,p. 68).
