Introduction light to follow the green path.

However, if
the light hits the black section, it is blocked
Ultraviolet and visible (UV/Vis) shortly to give allowance for any current
absorption spectroscopy measures the generated by the detector in the absence of
attenuation of a beam of light when it light.
passes through or interacts with an
absorbing species. Attenuation can be
brought about by absorption, scattering,
reflection, or interference. To produce
accurate quantitative measurements, the
attenuation measured should come only
from the absorption caused by the
absorbing species (Tissue, 2012).

Figure 2. Rotating disc components. Retrieved

from
https://www.chemguide.co.uk/analysis/uvvisible/
spectrometer.html

The sample and reference cells contain the

Figure 1. Basic components of a double beam
spectrometer. Retrieved from
https://www.chemguide.co.uk/analysis/uvvisible/
spectrometer.html
Illustrated in Figure 1 is a simple
double beam spectrometer. A UV/Vis
spectrometer works in a range from about
190 nm (near UV) to 900 nm (very near
infrared) (Tissue, 2012). The light source
used is a combination of two – a deuterium
lamp for the UV region and a
tungsten/halogen lamp for the visible region
– which is focused onto a diffraction grating
to scatter light of different wavelengths. The
slit then permits light of a very small range
of wavelengths to enter and hit the rotating
disc, which typically has three components,
namely, the transparent, mirrored, and black
sections (see Figure 2). If the light hits the
transparent section, it will follow the red
path shown in Figure 1. On the other hand,
hitting the mirrored section will cause the
solution with the investigated compound
and pure solvent, respectively. Conversion
of the received light into a current is done by
the detector and finally, the chart recorder
shows the output plot of absorbance against
wavelength (Clark, 2006).
For light to be absorbed in the
UV/Vis spectral range, the molecule must
possess chromophores which are groups,
such as pi bonds or lone pairs, that absorb
light. The wavelength of absorption, λmax ,
would depend on the presence of these
light-absorbing groups (Clark, 2007).
The absorbance of light by a sample
is directly related to the distance travelled
by the light through the sample and to the
concentration of the absorber according to
Beer-Lambert law, which is expressed as
A=ε ×b × c
where A is the absorbance measured at
wavelength λ , ε is the wavelength-
dependent molar absorption coefficient,
 b is the path length through the sample,
and c is the absorber concentration.
Relationships with absorbance are
observed to be linear, thus allowing
accurate concentration measurements of an
absorbing substance in a provided sample
(Laqua et al., 1988). However, the Beer-
Lambert law is not reliable over certain
concentration ranges – i.e. higher
concentration. Hence, it is more accurate to
measure the concentration via direct
calibration method, wherein solutions of
known concentration of the investigated
compound are prepared and measured at
the absorption maximum. Absorbance is
plotted against concentration and from the
graph, the unknown concentration of a
sample can be determined given its
absorbance (Clark, 2007).
particularly caffeine, and sodium benzoate
which will be determined as benzoic acid
(BA). Caffeine is a white crystalline xanthine
alkaloid which is added to soft drinks as a
flavor additive. On the other hand, BA is a
white crystalline solid and its sodium salt,
benzoate, is used as a food preservative. As
shown in Figures 3 and 4, the structures of
caffeine and benzoic acid, respectively, are
aromatic and hence they can absorb light in
the UV/Vis region (Gami & Sarasan, 2016;
McDevitt, Rodriguez, & Williams, 1998;
Patil, 2012).

UV/Vis analysis usually involves a

straightforward sample preparation. For
absorbing species that can be dissolved, Figure 3. Structure of caffeine. Retrieved from
measurements are done in solution using a http://www.chem.ucla.edu/
suitable solvent or pH buffer. To make sure
that the absorber is in the same form both in
standard and sample solutions, pH effects
and complexation should be monitored.
Non-absorbing substances can also be
quantified by simply adding a reagent with
which it can form an absorbing complex Figure 4. Structure of benzoic acid. Retrieved
(Dean, 1995). from http://www.chem.ucla.edu/

In quantitative measurements, it is Hence, the specific objective of this study

necessary that only a few absorbing species
in a sample are present because UV/Vis
spectra typically involve broad bands.
Samples with many absorbers are to
undergo separation first prior to quantitative
analysis. Likewise, any particles present in
the sample should be removed via
centrifugation or filtration to prevent light
scattering which interferes with absorbance
measurement (Tissue, 2012).
In this study, it was aimed to analyze
commercial soft drink components,
was to quantify the concentration of caffeine
and benzoic acid in 7UP via UV/Vis
absorption spectroscopy.
Methodology
A. Standard Solution Preparation
Stock solutions of BA and caffeine
having a concentration of 100 ppm were
made. In separate 50-mL beakers, 10.0 mg
of BA and 10.7 mg caffeine were weighed
and dissolved in a small amount of water.
The solutions were then transferred
quantitatively to separate 100-mL volumetric
flasks and diluted to mark with dH2O.
From the stock solutions, working
standard solutions were prepared in
duplicates. Five clean, 50-mL volumetric
flasks were secured for each stock solution.
By means of a pipettor, 1, 2, 3, 4, and 10
mL of the BA stock solution were transferred
separately to the flasks to make 2, 4, 6, 8,
and 20 ppm of BA solution respectively.
Similarly, 4, 8, 12, 16, and 20 ppm caffeine
solutions were made by using 2, 4, 6, 8, and
10 mL of the caffeine stock solution. To all
solutions, 10 mL of 0.10 M HCl was added
after which the flasks were filled to mark
with dH2O.
B. Sample Preparation
Approximately 20 mL of 7UP was
placed in a beaker and subsequently
warmed on a hot plate to expel CO2. The
warm sample was passed through a filter
paper to remove any present particles and
then cooled to room temperature. Using a
pipettor, 2 mL and 4 mL of the sample were
transferred to separate 100-mL volumetric
flasks. To both volumes, 10 mL of 0.10 M
HCl was added after which the flasks were
filled to mark with dH2O.
C. Absorbance Measurements
Absorbance measurements were
supposed to be done using a UV-Vis
spectrometer via wavelength scan of both
standard solutions from 350 to 210 nm to
obtain the λmax for both BA and caffeine,
followed by the measurement of
absorbance of each standard and of the two
sample solutions at both maximum
wavelengths with dH2O as the blank
sample. Instead, experimental data were
given by the instructor for analysis.
Plots of absorbance against
concentration of the standard solutions for
each of the two wavelengths were
constructed and using direct calibration
method, the concentration of BA and
caffeine in the original soft drink was
calculated.
References
Clark, J. (2006). A double beam UV-visible
absorption spectrometer. Retrieved from
https://www.chemguide.co.uk/analysis/uvvisi
ble/spectrometer.html
Clark, J. (2007). Using UV-visible
absorption spectra. Retrieved from
https://www.chemguide.co.uk/analysis/uvvisi
ble/analysis.html
Clark, J. (2007). UV-visible absorption
spectra. Retrieved from
https://www.chemguide.co.uk/analysis/uvvisi
ble/theory.html
Dean, J. A. (1995). Analytical chemistry
handbook. New York: McGraw-Hill.
Gami, R., & Sarasan, G. (2016). A
comparative study of concentration of
caffeine and benzoic acid in various soft
drink samples. International Journal of
Science and Research, 5(8), 1655-1659.
Laqua, K., Melhuish, W. H., & Zander, M.
(1988). Nomenclature, symbols, units, and
their usage in spectrochemical analysis –
VII. Molecular absorption spectroscopy,
ultraviolet and visible (UV/VIS). Pure
Applied Chemistry, 60, 1449-1460.
McDevitt, V. L., Rodriguez, A., & Williams,
K. R. (1998). Analysis of soft drinks: UV
Spectrophotometry, Liquid Chromatography,
and capillary electrophoresis. Journal of
Chemical Education, 75(5), 625.
doi:10.1021/ed075p625
Patil, P. N. (2012). Caffeine in various
samples and their analysis with HPLC - A
review. International Journal of
Pharmaceutical Sciences Review and
Research, 16(2), 76-83.

Tissue, B. M. (2012). Ultraviolet and visible

absorption spectroscopy. In E. N. Kaufmann
(Ed.), Characterization of materials. USA:
John Wiley & Sons, Inc.