Introduction

Fluorescence is the emission of light from a substance that has absorbed electromagnetic
radiation or other light sources. In other word, it is a luminescent without a change in spin
multiplicity. By the action of molecules being excited to higher energy level, the fluorescence
is produced. It can be described energy being absorbed and excited to higher energy level,
followed by vibrational relaxation, internal crossing, vibrational relaxation which leads to
fluorescence Normally, most substance absorbs energy from uv-region and then emits light
within visible region. In order to detect the level of fluorescence, an electromagnetic
spectroscopy called as fluorescence spectroscopy is used. It uses a beam of light which is
passes through a sample that have a selected analyte, which is then excited by the
absorption of the energy, thus causing them to emit light. For this kind of spectroscopy, a
certain analyte must have the following characteristics; having benzene ring structure or
conjugated double-double bond as it is needed for the emission of fluorescence. Thus the
sample used in this experiment has the proper characteristic as latter and as such, quinine in
energy drinks are used as the analyte for easy to obtain and measured using FES device
due high quantum yield.

Purpose

To provide correlation of standards and to detect the concentration of quinine in a given

energy drink (Livita).
Procedure

Standard Preparation

About 10mg of quinine sulphate is obtained which is dissolved in 1 litre of distilled water in a
dark container to produce 10ppm solution. The 0.1M sulphuric acid solution is made from
stock solution of 18M in 250ml volumetric flask. The standards are prepared in the following
table in 25mL volumetric flask:

Standards (g/mL) Volume of 10ppm Volume of distilled Volume of 0.2N

quinine (mL) water (mL) sulphuric acid (mL)
0.2 0.5 12.0
0.4 1.0 11.5
0.6 1.5 11.0
0.8 2.0 10.5 12.5
1.0 2.5 10.0
Total volume of each standard=25.0mL

Sample Preparation

The sample is prepared from 0.5mL of Livita energy drink with the addition of 12.0mL of
distilled water and 12.5mL of 0.1M sulphuric acid in 25mL volumetric flask. It is then tested
with UV-vis.

Data
 Ex. Wavelength (nm) 238
Em. Wavelength (nm) 441
Ex. Slit (nm) 2.5
Em. Slit (nm) 2.5
Integration Time (s) 1.00
Correlation Coefficient 0.9992

Standard Conc*Fact Intensity

Std 1 0.200 218.453
Std 2 0.400 416.346
Std 3 0.600 611.669
Std 4 0.800 800.995
Std 5 1.000 999.999
Sample (Livita) 0.061 61.680

Discussion

The main principle of fluorescence is the emission of radiation after the absorption of light
under condition of the emission does not involves spin change. The experiment starts with
the preparation of standards in unit of microgram per millilitre; 0.2, 0.4, 0.6 0.8 and 1.0. The
stock is prepared by dissolving 10mg of quinine sulphate in 1 litre of distilled water in dark
volumetric flask which blocks out UV light from being absorbed by the analyte which can
cause run off during the reading of the fluorescence level. These standards are run to
provide a correlation between the standards and the sample in term of coefficient, given
0.9992 and the graph, which generated by the system. From these, the concentration of the
sample can be determined. The sample, is prepared by pipetting 0.5 mL of tonic water (Livita
energy drink), into a dark 50mL volumetric flask, again to block out any stay UV-light. It is the
added with 12.0mL distilled water and 12.5mL of 0.1M sulphuric acid. The prepared sample
is tested with FES machine to determine the concentration of quinine within the tonic water.
The given concentration is at 0.061mMol per litre. The reason why quinine is mixed with
sulphuric acid is the fluorescence of quinine given out is much stronger in the presence of
acid.

Conclusion:

The concentration of quinine presence in Livita tonic water is at 0.061mMol per litre.

References:

1. Picture, http://www.chemistry.adelaide.edu.au/external/soc-rel/content/images/fluo-
sch.png
2. Notes on Fluorescence, CHM 312
3. Explanation on Fluorescence, http://en.wikipedia.org/wiki/Fluorescence_spectroscopy