SSK 3213 Analytical Chemistry II

TUTORIAL 3 FLUORESCENCE AND PHOSPHORESCENCE

1. Define the following terms:

a. Singlet excited state and triplet excited state
b. Fluorescence
c. Phosphorescence
d. Internal conversion
e. External conversion
f. Intersystem crossing

2. Discuss briefly how measurement of fluorescence emission is carried out.

3. Draw a schematic diagram of fluorescence instrumentation.

4. Why is it imperative that the optical paths for the source and detector are
positioned at an angle of 90o?

5. Discus the advantages of using fluorescence spectrometric method of

analysis.

6. Explain why the wavelength of fluorescence emission must always be

greater than the excitation wavelength?

7. What are the principle differences between fluorescence and

phosphorescence?

8. A sample of tonic water was analyzed using fluorescence spectroscopy for

its quinine content at an excitation wavelength of 350 nm and emission
wavelength of 450 nm. 1 mL sample of tonic water was diluted to 100 mL
with 0.05 M H2SO4 and gave an emission intensity of 8.4. Determine the
concentration of quinine in the sample. The following solutions of quinine
were used as standard and the fluorescence emissions were as recorded.

Concentration, ppm Fluorescence Intensity

1.00 12.0
0.80 9.5
0.60 7.2
0.40 4.8
0.20 2.5

9. Thorium can be determined by fluorescence spectrometry using 1-amino-

4-hydroxyanthraquinone, by means of standard addition method. A certain
volume of ThO2+ 2.00 × 10-5M was added to a series of 0.250 g sample
 which was then diluted to 100 mL with distilled water. The fluorescence
intensities were as follows:

Added ThO2+ 0.0 1.0 3.0 5.0 7.0 10.0

Intensity 23.7 31.0 45.6 60.0 74.7 96.3

Calculate the concentration of Th in the sample.

10. Aspirin tablets were analyzed using fluorescence spectroscopy. The

salicylic acid content was analyzed at 450 nm. 100 mg tablet was diluted
to 100 mL with 5% acetic acid-chloroform solution and gave an emission
intensity of 218. The following are the intensities of the standard solutions.
Calculate the concentration of salicylic acid in the sample.

ppm 0.0 2.0 4.0 6.0 8.0 10.0

Intensity 4 69 134 199 265 329