SWISS GERMAN UNIVERSITY

INORGANIC AND ORGANIC CHEMISTRY

LABORATORY REPORT
Subject
Lecturer
Instructor
Faculty/Class
Date of Experiment
Date of Lab. Report
Semester
Time of Experiment

: Inorganic and Organic Chemistry Laboratory

: Dr. rer. nat. Filiana Santoso, Mr. Hery Susanto M.Si
: Mr. Tabligh Permana, Mr.Hery Sutanto M.Si, Ms. Sylvia Yusri, S.Si
: Life Science/LS 2A
: 25 March 2014
: 8 April 2014
: 2
: 14.00 17.00 p.m

Experiment:
Name:

Campus BSD City

Bumi Serpong Damai
Tangerang 15321 Indonesia

Principle of Spectroscopy

Kristania Hadhiwaluyo
Chita Sakina Putrianti
Elias Harmanto

Tel.
Fax.

+62 21 537 6221

+62 21 537 6201

sgu.info@sgu.ac.id
www.sgu.ac.id

I.

II.

Objectives

To understand the principle of spectroscopy.

To determine the concentration of an unknown sample of solution

Theoretical Background

Spectroscopy was originally the study of interaction between radiation and matter as a function of
wavelength. Spectroscopy referred to the use of visible light dispersed according to its wavelength.
Over a certain range of wavelength, every chemical compound absorbs, transmits, or reflects light,
spectrophotometry is a measurement of how much a chemical substance absorbs or transmits and
a spectrophotometer is an instrument that measures the amount of the intensity of light absorbed
after it passes through sample solution. With the spectrophotometer, the concentrations of a
substance (the amount of a known chemical substance) can also be determined by measuring the
intensity of light detected. It can be classified into three different types, depending on the range of
wavelength of light source:

UV-visible spectrophotometer: uses light over the ultraviolet range (185 - 400 nm).
UV spectroscopy is used for fluids, and even solids. Cuvettes, only made of quartz, are
used for placing the samples.

Visible Light (400-700 nm): Visible spectrophotometers, which are used in this
experiment, can use incandescent, halogen, LED, or a combination of these sources
and these spectrophotometers vary in accuracy. Plastic and glass cuvettes can be used
for visible light spectroscopy

IR spectrophotometer: uses light over the infrared range (700 - 15000 nm) of
electromagnetic radiation spectrum. IR spectroscopy helps to study different structures
of molecules and their vibrations. Different chemical structures vibrate in different ways
due to variation of energy associated with each wave length.

In experimental ways where it is used to identify substances through the spectrum emitted from
or absorbed by them, the basic function of a spectrometer is to take in light, break it into its
spectral components, digitize the signal as a function of wavelength, and read it out and display it
through a computer. The first step in this process is to direct light through a fiber optic cable into
the spectrometer through a narrow aperture known as an entrance slit. The slit vignettes the light
as it enters the spectrometer. In most spectrometers, the divergent light is then collimated by a
concave mirror and directed onto a grating. The grating then disperses the spectral components of
the light at slightly varying angles, which is then focused by a second concave mirror and imaged
onto the detector. Once the light is imaged onto the detector the photons are then converted into
electrons, which are digitized, and readout through a USB (or serial port) to a computer. The

software then interpolates the signal based on the number of pixels in the detector and the linear
dispersion of the diffraction grating to create a calibration that enables the data to be plotted as a
function of wavelength over the given spectral range.
Spectrophotometry works on a very basic
principle that every atom is capable of absorbing
light. Beer and Lamberts law, which is the basis
of spectrophotometry, stated the linear
relationship between absorbance and
concentration, meaning concentration is directly
proportional to absorbance of that certain atom
and inversely proportional to transmittance. It is
because the more concentrated that solute is in
the solution the more light should be absorbed by
that solution (absorbance increase) and the less
light will pass through that solution
(transmittance). The light itself is the result of
electrons moving between defined energy levels
in an atom. When something excites an atom an
electron may absorb the energy, boosting it up to
a higher-level shell. The boost is short-lived, and the electron immediately falls back down to the
lower level, emitting its extra energy in the form of an electromagnetic energy packet called a
photon. The wavelength of the photon depends on the distance of the electrons fall. Some
wavelengths, such as radio waves, are invisible to our
eyes however photons with wavelengths in the visible
spectrum form all the colors that we can see.
A substance will appear a particular color because it
absorbs visible light corresponding to its complimentary
color, which is the color that is directly opposite absorbed
light on the color wheel. However, the color of an object
that we see actually not contained within the object,
instead it is the light that is any visible light that strikes the object and becomes reflected or
transmitted to our eyes will contribute to the color appearance of that object. Finally the
relationship between absorbance and transmittance of Beer Lamberts law and the reason why
different concentration gives different intensity of colors will be explain further in the report.

III.

Equipment and Materials

IV.

Equipment:
-

Volumetric flask, 2, 100 cm3

Volumetric flask, 2, 25 cm3

Volumetric flask, 1, 10 cm3

Graduated pipette, 25 cm3

Graduated pipette, 10 cm3

Graduated pipette, 5 cm3

Erlenmeyer flask, 100 cm3

Bulb, 3

Test tube, 5

Rack of test tube

Pipette

Glass Beaker

Cuvette, 5

Spectrophotometer

Computer

Erlenmeyer Flask

Materials:
-

H2O(l), distilled water, 205 cm3

H2SO4(l), concentrated sulfuric acid, 2 cm3

K2CrO4(l), 0.1M potassium chromate, 98 cm3

Procedures

1. Step 1: Preparation of Solutions

1. The (grandparent) solution, 100 cm3 of 0.1 M K2CrO4(l) (potassium
chromate), was prepared by adding 50 cm3 of H2O(l) (distilled water), 2 cm3
of H2SO4(l) (sulfuric acid) and 48 cm3 of K2CrO4(l) (potassium chromate).
2. Potassium chromate solution from the 1st step was diluted into 100 cm3 of
0.01 M K2CrO4(l) and considered to be another parent solution by adding 10
cm3 of 0.1 M K2CrO4(l) and 90 cm3 of distilled water, H2O(l).
3. The solution produced in step 2 was then further diluted to create 4
different solutions with different molarity:
i. 7.5 cm3 of 0.1 M K2CrO4(l) was taken into 10 cm3 volumetric flask and
added with 2.5 cm3 of distilled water to make solution of 0.0075 M
K2CrO4(l).

ii. 12.5 cm3 of 0.1 M K2CrO4(l) was taken into 25 cm3 volumetric flask
and added with 12.5 cm3 of distilled water to make solution of 0.005
M K2CrO4(l).
iii. 25 cm3 of 0.1 M K2CrO4(l) was taken into 100 cm3 volumetric flask and
added with 75 cm3 of distilled water to make solution of 0.0025 M
K2CrO4(l).
iv. 2.5 cm3 of 0.1 M K2CrO4(l) was taken into 25 cm3 volumetric flask and
added with 22.5 cm3 of distilled water to make solution of 0.001 M
K2CrO4(l).

The
Grandparent
solution

The solutions that were used in this experiment including the unknown
sample

4. Solution of 0.001 M K2CrO4(l), 0.0025 M K2CrO4(l), 0.005 M K2CrO4(l), and

0.0075 M K2CrO4(l) was put into 4 different test tubes with the same height.
5. Unknown solution was also placed into a test tube with the same height with
other solutions.
6. Five different solutions were observed and sorted according to their color.

2. Measuring Solutions in Spectrometer

1. Each solution was put into 5 different cuvettes and was labeled with label of
different colors.

2. The cuvettes were put onto the spectrophotometer that was connected to
the computer shown below.

3. The concentration and the absorbance was recorded into the data table
shown below:
Standard Curve Data
No

Concentration [M/L]

Absorption [A]

Error [A]

Used

0.0075

0.026

0.001

Yes

0.0050

0.016

-0.001

Yes

0.0025

0.039

0.031

Yes

0.0010

0.004

0.001

Yes

Unknown Sample Data

Sample

Dilution Factor

Ordinate [A]

Concentration [M/L]

Sample

0.013

0.00383

V.

Observation (Data)
Standard Curve Data
No

Concentration [M/L]

Absorption [A]

Error [A]

Used

0.001

0.190

-0.027

Yes

0.0025

0.528

-0.015

Yes

0.005

1.099

0.013

Yes

0.0075

1.630

0.000

Yes

Unknown Sample Data

VI.

Sample

Dilution Factor

Ordinate [A]

Concentration [M/L]

Unknown Sample

0.401

0.00185

Discussion

Graph of Concentration versus Absorbance of the

Unknown Sample
= 440 nm 0.400 A

Concentration of The Sample

y
= 217.2881x
0.401 = 217.2881x
x
= 0.00185
From the value of the absorbance, %transmittance also can be determined:
Absorbance (A)
= -log (%transmittance/100)
Transmittance
=
Transmittance
=
Transmittance
=
So, the %transmittance at 440 nm is 39.7%

ANALYSIS
Why yellow?
Primarily light that every atom in a substance absorb the result of electrons moving
between defined energy levels in an atom, when something excites an atom an electron
may absorb the energy, boosting it up to a higher-level shell but since a high energy level
is considered unstable for the electrons, they immediately falls back down to their original
level while emitting its extra energy in the form of an electromagnetic energy packet called
a photon. The wavelength of the photon depends on the distance of the electrons fall,
some wavelengths, such as radio waves, are invisible to our eyes but wavelengths in the
visible spectrum (light with a range of 400-700 nm) form all the colors that we can see

The light that we see may exist in many different colors, however the color that we see is
the color that is not contained within the object, instead it is the light that is any visible
light that strikes the object and becomes reflected or
transmitted to our eyes. Molecules are able to absorb
energy of different wavelength from the visible light
spectrum and then emits the complementary color of
the color its absorbs. In the case of this experiment,
the molecules inside this solution has absorbed the
purple region of the visible light spectrum with the
wavelength of 440 nm thus allowing them to emit
yellow which is the complementary color of blue (seen in
the color wheel on the right).
However, each different solution absorbs light in different amount of wavelength
depending on its concentration. It is seen from the diagram below where the solution are

placed inside the cuvette in order of decreasing color intensity that although they all emit
the same yellow color but the more concentrated one has a higher color intensity than
those which are less concentrated. This is because in a more concentrated solution there
are more molecules that are able to absorb the purple region of the visible light spectrum
compared to those, which are less concentrated, and therefore has lesser amount of
molecules. Finally this qualitative observation is proven further from the strong positive
correlation (value that is close to 1) that is represented as continuous increasing trend of
the standard graph of absorbance versus concentration that is obtained from the
spectrophotometer (equipment that we used to analyze those samples). Hence the
continuous increasing trend of the standard graph eventually verified the Beer and
Lamberts law which stated the linear relationship between concentration and absorbance
of a substance.

Graph of Concentration versus Absorbance of the

Unknown Sample
= 440 nm 0.400 A

Predicting the concentration of the unknown sample

In addition, this qualitative observation also enables us to predict the concentration of the
unknown sample, which is labeled with a pink label. Qualitatively, by comparing its color
with other known sample, we can be sure that the concentration of the unknown sample
lies between 0.001 to 0.0025 M. It is due to its color intensity that is not as pale as 0.001
M but not too dark as the 0.0025 M solution. This is also proved from our sample-analysis
that is conducted with the spectrophotometer that the solution with concentration of 0.001
to 0.0025 M (the predicted range) has absorbance value that range between 0.190 to
0.528 whereas the unknown sample of solution has the concentration of 0.0185 M with
absorbance of 0.400. The concentration and absorbance value that this unknown sample
has is located within the predicted range, therefore qualitatively we see that the color of
this unknown sample is similar with the other solution with the concentration of 0.001 to
0.0025 M. Additionally, in terms of absorbance of the solution itself, the ones with
concentration of 0.001 to 0.0025 M should have the similar value of absorbance as the
unknown sample of solution (around 400nm) and absorb the similar region of purple region
of the visible spectrum with different intensity depending on the amount of molecules
inside the solution that absorbs the light (more concentrated / contains more molecule =
more absorbance).
Absorbance and transmittance
The percent transmittance is defined as the amount of light that passes through a sample.
In terms of its relationship with each other, absorbance is inversely proportional to
transmittance. It is illustrated in the diagram below that lower % transmittance
corresponds to the higher absorbance therefore if all the light passes through a solution
without any absorption, then absorbance is zero, and percent transmittance is 100%. If all
the light is absorbed, then percent transmittance is zero, and absorption is infinite.

In this experiment the percent transmittance of the unknown sample is calculated using
the formula: Absorbance = -log (percent transmittance/100) which finally give us to value
of 39.7%. Finally by looking back at the diagram above and the data gained from the

spectrophotometer we can conclude that if the unknown sample of solution has a percent
transmittance of 39.75%, this would mean that the unknown sample will absorb light
around 0.4-0.5 which is also the exact value of absorbance (0.400).
VII.

Conclusion
In our experiment, all the samples of the solution including the unknown have the similar
yellow color. It is because they absorb the similar value of wavelength in the purple region
of the visible spectrum and emits the color yellow, which is the complementary color of
purple. Secondly, from analyzing the entire samples we successfully verified the Beer and
Lamberts law about the linear relationship between concentration and absorbance of a
certain solution. It is because proven qualitative and quantitatively from our experiment
that as the concentration of our samples increase from 0.001, 0.0025, 0.005, to 0.075M it
contains higher amount of molecules that can absorb the purple region of the visible light
therefore increase the value of absorbance that it has. In addition our experiment also
proved the inversely relationship between absorbance and transmittance of a certain
solution, all the light passes through a solution without any absorption, then absorbance is
zero, and percent transmittance is 100% so as it absorb more light it transmit less amount
of light. More over, our experiment also concluded that spectrophotometry can help us to
identify the identity of an unknown sample quantitative and qualitatively. Qualitatively, we
can use our eyes in determining the range of concentration and wavelength where our
unknown sample belong by looking and comparing its color with the other samples. Finally
with the spectrophotometer we can calculate the exact value of both absorbance and
transmittance, the concentration and the exact value of wavelength of the unknown
solution itself and finally verify our prediction.

VIII.

References

Sutanto, Hery, and TablighPermana. Inorganic and Organic Chemistry 1 Laboratory

Manual.Tangerang: Swiss German University, 2013. Print.

"Definition of Percent Transmittance?." - Ask.com. N.p., n.d. Web. 6 Apr. 2014.
<http://www.ask.com/question/definition-of-percent-transmittance>.
"How Does a Spectrometer Work?." BW Tek. N.p., n.d. Web. 3 Apr. 2014.
<http://bwtek.com/spectrometer-introduction/>.
"How do atoms emit light?." How It Works Magazine. N.p., n.d. Web. 6 Apr. 2014.
<http://www.howitworksdaily.com/science/how-do-atoms-emit-light/>.
"Light Absorption, Reflection, and Transmission." Light Absorption, Reflection, and Transmission.
N.p., n.d. Web. 6 Apr. 2014. <http://www.physicsclassroom.com/class/light/u12l2c.cfm>.
"Montessori Muddle." Montessori Muddle RSS. N.p., n.d. Web. 6 Apr. 2014.
<http://montessorimuddle.org/2012/02/01/emission-spectra-how-atoms-emit-and-absorblight/>.
Reeves, Kimbrough. "Beers Law." University of North Carolina Wilmington. Version 2001. N.p.,
n.d. Web. 6 Apr. 2014.
<http://uncw.edu/chem/Courses/Reeves/OnLineLabs/beerslaw/beerslaw_intro.htm>.
Sheffield Hallam University.2000. Beers
Law.http://teaching.shu.ac.uk/hwb/chemistry/tutorials/molspec/beers1.htm
"SPECTROPHOTOMETRIC MEASUREMENTS TECHNIQUES FOR FERMENTATION PROCESS."

University of Oradea, Romania. N.p., n.d. Web. 4 June 2014.

<http://imtuoradea.ro/bioethanol/UV.VIS.theory.pdf>.
"Spectroscopy: Principles, Theory, Techniques and Applications." Metameso.org. N.p., n.d. Web. 4
June 2014. <http://metameso.org/nri/library/free-books/SpectroscopyOvrview.pdf>.