FACTORS AFFECTING FLUORESCENCE
Chandramouli R
INTRODUCTION
CONJUGATION SUBSTITUTION GROUPS
ELECTRON DONATING GROUPS ELECTRON WITHDRAWING GROUPS AROMATIC GROUPS ALIPHATIC AND ALICYCLIC CARBONYL GROUPS UNSUBSTITUTED AROMATIC HYDROCARBONS HETEROCYCLICS HALOGENS CARBONYL GROUP
RIGID STRUCTURE VISCOSITY TEMPERATURE POLARITY OF THE SOLVENT DISSOLVED OXYGEN PH
QUENCHING LIGHT ADSORPTION
CONJUGATION
The greater the absorption of a molecule, the more intense is its luminescence. Thus molecules having conjugate double bonds or bonds can be studied with the help of fluorescence. If there is no conjugation or bond, there will be no absorption.
SUBSTITUTION GROUPS
Electron donating groups: Like -NH2, -OH etc it enhance the fluorescence intensity. Electron withdrawing groups: Like -COOH,-NH2 etc reduce (or) even destroy the fluorescence. Groups like SO3H,-NH4+, Alkyl do not have much effect on both fluorescence and phosphorescence.
Aromatic groups: The most intense and most useful fluorescence has been observed in this groups with low energy -* transition level Aliphatic and Alicyclic carbonyl structures: It is are highly conjugated double bond structures have also been found to exhibit fluorescence.
Unsubstituted aromatic hydrocarbons:
These are fluoresce in solution. The quantum efficiency usually increase as the number of rings and their degree of Condensation increases
Heterocyclic:
Like pyridine, furan, thiophene and pyrrole dont exhibit fluorescence.
The lowest energy electronic transition involves n-* system, Which rapidly converts to the triplet state hence prevent the fluorescence. But, Heterocyclics with fused rings, exhibit fluorescent behaviour. Fusion with Benzene rings to a Heterocyclic nucleus it increase the molar absorptivity of absorption peaks The life time of an excited state is shorter in fused systems. So, the compounds Quinoline, Isoquinoline, indole have fluorescence.
Halogens:
The fluorescence has been found to decrease with increasing atomic number of the substituted halogen. Due to the heavy atom effect which increase the rate of intersystem crossing to the triplet state.
S.No SUBSTITUENT EFFECT ON 1 2 3 4 5 6 7 8 9 Alkyl COOH,CHO, COOR,COR OH,OMe,OEt CN NH2,NHR,NR2 NO2,NO SH SO3H F, Cl, Br, I No effect No effect No effect
EFFECT ON INTENSITY Slight or
No effect
CARBONYL GROUP In a aromatic ring substitution-COOH or C=0 is inhibit the fluorescence In this compounds the energy in the n, is less than in the -* system.
VISCOSITY
Increasing the viscosity increase the fluorescence. Because the frequency of collisions is decreased. Decreasing the viscosity decrease the fluorescence.
RIGID STRUCTURE
Fluorescence is favoured in molecules that possess rigid structures. FLUORENE BIPHENYL
These is due to the increased rigidity furnished by the methylene group to Fluorene. Fluorescing dyes enhance the frequency of emission when absorbed on a solid surface. Increase in fluorescence of some organic chelating agents when they complexed with a metal ions. It is greater in the zinc complex
8-HYDROXY QUINOLINE
TEMPERATURE
Increase in temperature leads to decrease in fluorescence. Because the frequency of collisions increase and quantum yield decreased Quantum yield= No of molecules that fluorescence/Total no of excited molecules.
POLARITY OF THE SOLVENT
Solvents containing heavy atoms or other solutes with such atoms in their structure decrease the fluorescence.
e.g.: carbon tetra bromide, ethyl iodide.
In polar solvents, The energy of the n-* transition is often increased. The energy of the -* transition is often decreased. In some cases, Shifts may be great enough to lower the energy of the - * process below that of the n-* transition and enhance fluorescence results
DISSOLVED OXYGEN
Oxygen decreases the fluorescence in two ways By photochemically induced oxidation of the fluorescent material. Quenching also occurs as a result of paramagnetic properties of molecular oxygen. It promote the intersystem crossing and conversion of excited molecules to the triplet state.
H P
Neutral or alkaline solution of aniline shows fluorescence, In visible region fluorescence disappears. Aniline shows blue fluorescence in the range of pH 5-13 when excited at 290nm. At lower pH, aniline exists as the aniline cation [pH<5] In highly alkaline media, At pH>13 as anion, neither anion over cation is fluorescent.
QUENCHING
Quenching is the decrease in flourecence intensity due to specific effects of constituents of the solution itself. These effects due to various factors like, Concentration pH Presence of specific chemical substance Temperature. Viscosity etc
TYPES OF QUENCHING:
Self Quenching Chemical Quenching Static Quenching Collisional Quenching
1. SELF QUENCHING: - At low concentration (g or ng/ml), linearity
is observed. - At high concentration (mg/ml) of the same substance, proportionate increase in flourescence intensity does not occur.
2.CHEMICAL QUENCHING:
In this type, quenching is due to various factors like, - Changing in PH - Oxygen - Halides and electron withdrawing groups - Heavy metals
3.Static Quenching:
This occurs because of complex formation. E.g: Caffine reducing the flourescent intensity of Riboflavin, by complex formation.
4.Collisional Quenching:
If collisions are increased, quenching takes place
ADSORPTION
-
Require very dilute solution only - 10-100 times weaker than the spectrometry - Adsorption of the fluorescent substance on the container walls
E.g: Quinine
LIGHT
Only Monochromatic light because the intensity will vary with wavelength
