CABRERA, ALLIAH ROME A.

BSCHEM 2

Chem 2122
Problem Set 10. Spectroflourometry

1.) Describe the principles of Phosphorescence. Why are phosphorescence measurements

frequently made at liquid nitrogen temperatures?
Ans. A type of photoluminescence called phosphorescence, which is related to fluorescence,
slowly releases the stored energy over time. Phosphorescent material is essentially "charged" by
being exposed to light. The energy is then held for a while before being gradually released. In
other area, the reason why phosphorescence measurements frequently made at liquid nitrogen
temperatures is to slow down the rate of radiationless transfer processes in order to increase
phosphorescence.

2.) Which compound in each of the pairs below would you expect to have a greater
fluorescence quantum yield? Explain.

Ans. Fluorescein has fused rings and a more rigid structure than phenolphthalein so
consequently Fluorescein has a higher quantum yield.

Ans. According to the figure, the double bond between the two nitrogen groups in compound
bis (o-hydroxyphenyl) hydrazine is absent, whereas the double bond between the two nitrogen
groups in compound o, o' - dihydroxyaxobenzene is in conjugation between the aromatic rings.
Resulting, the fluorescence quantum yield of o, o' - dihydroxyaxobenzene is higher.
3.) Figure 1 spectrum was obtained for a liquid with the empirical formula of C 3H6O. Interpret
the IR spectrum. What is the possible structure of the compound?

Ans. 2950- CH Alipatic stretch, 3350- OH stretch, 1680-C=C stretch

In order to guarantee the presence of C=C, an -OH group, and aliphatic -C-H stretching in the
molecule with the given empirical formula.
Possible structure of the compound: CH2=CH-CH2-OH

4.) Figure 2 spectrum is that of a high- boiling point liquid having the formula of C9H10O.
Identify the compound and interpret the IR spectrum.

Ans. The absorption peak at 3000 cm-1 indicates the presence of aromatic C-H stretching. The
absorption peak at 1700 cm-1 indicates the presence of ketone. The absorption peak at 1350
cm-1 indicates the presence of CH2. The C-H absorption at 2850 cm-1 to 3000 cm-1 indicates
the presence of CH3. The absorption peak at 745 cm-1 and 705 cm-1 indicates five adjacent
hydrogen atoms on an aromatic ring.
5.) A bottle of tonic water is to be analyzed for its quinine content by fluorescence spectrometry,
with excitation at 350 nm and emission intensity measured at 450 nm. One milliliter (1 mL) of
tonic water is diluted to 100 mL with 0.05 M H2SO4; its emission intensity is 8.44 (arbitrary units).
A series of quinine standards, in 0.05 M H2SO4, is prepared and the emission intensities measured
in parentheses): 100 ppm (293 units), 10.0 ppm (52.3), 1.00 ppm (12.0),0.100 ppm (1.26), 10
ppb (0.158), and 1.0 ppb (0.015). The emission intensity of 0.05 M H2SO4 is negligible. Plot the
calibration curve for quinine fluorescence and determine the quinine content of the original tonic
-water sample.

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