Module 1:

Exercise
Calligraphers United 1

DETERMINATION OF THE
CONCENTRATION OF AN UNKNOWN
SOLUTION USING A COLORIMETER or
UV-VIS SPECTROMETER (BEER’S LAW)

Objectives
At the end of the exercise, the students are expected to:
Examine the relationship between the absorbance and concentration of a
copper (II) sulfate solution, and
Determine the concentration of an unknown copper (II) sulfate solution
using a colorimeter or UV/VIS Spectrophotometer.

Data

DELDIO, Veronica Lourds A. | BSP2E

Solution

DELDIO, Veronica Lourds A. | BSP2E

Results

We determined the concentration of the sample, which is

approximately 0.36 M. this was determined by using the
absorbance of the sample and going from the point of interception
on the slope.

Conclusion
In conclusion, the graph that was used to show its direct
relationship for the linear slope is created.

DELDIO, Veronica Lourds A. | BSP2E

 Module 1:
Exercise 1
Pre-Task
Define and state the principle of each instrument: Define and
state the principle of each instrument:

Question 1
A. Colorimetry
Definition: Colorimetry is the field of determining the concentration of a colored
compound in a solution. Calorimetry was also known as FILTER PHOTOMETER, is
an analytical machine that acts as the tool to quantify the concentration of a
solution by measuring the absorbance of a specific wavelength of light.

Principle: This working principle of the colorimeter is based on Beer-Lambert's

Law which states that the amount of light absorbed by a color solution is
directly proportional to the concentration of the solution and the length of a
light path through the solution.

A. Spectometry
Definition: Spectrophotometry is a method to measure how much a chemical
substance absorbs light by measuring the intensity of light as a beam of light
passes through the sample solution. The basic principle is that each compound
absorbs or transmits light over a certain range of wavelength.

Principle: The spectrophotometer technique is to measure light intensity as a

function of wavelength. It does this by diffracting the light beam into a spectrum
of wavelengths, detecting the intensities with a charge-coupled device, and
displaying the results as a graph on the detector and then on the display devic.

DELDIO, Veronica Lourds A. | BSP2E

Question 2
Differentiate Absorption methods from Colorimetric methods.

Absorption techniques are as often as possible utilized as distinguishing proof

tests for drug substances. Albeit every one of the three locales of the
electromagnetic range is utilized, the UV and IR areas are utilized most habitually.
While using the UV area with the end goal of distinguishing proof, the test
generally comprises of at least one of the accompanying three systems: (1)
assurance of absorbance esteems at two frequencies, trailed by computation of
the proportion and examination with the worth given in the authority abstracts;
(2) assurance of the absorptivity estimations of the authority substance
and a relating reference standard and correlation of the outcomes; or (3)
correlation of the reference standard range with an example range.

DELDIO, Veronica Lourds A. | BSP2E

 Module 1: Exercise 1
Check-in
Calligraphers Activity
United

Draw and label parts of

colorimeter and UV/VIS
Spectrophotometer

Parts of Colorimeter and UV/VIS Spectrophotometer:

1. LIGHT SOURCE- serves as the light source or the radiation source. it is usually a
hydrogen or deuterium lamp and tungsten for visible measurements.
2. COLLIMATOR – This is used to produce a parallel beam of rays or radiation.
3. MONOCHROMATOR (PRISM OR GRATING) – This is used to select a narrow band of
wavelength to pass through the sample cell, an important property of a
monochromator is bandpass or bandwidth.
4. WAVELENGTH SELECTOR (SLIT)- used to focus or collect the radiation of light
into a single beam or ray, thus, preventing unwanted light or stray light.
5. SAMPLE CELL (CUVETTE) – This is used to hold the working solution.
6. DETECTOR (PHOTOCELL) – This is used to change the radiation
transmitted from the UV-VIS spectrophotometer into a current or voltage
for the readout device to use.
7. DIGITAL DISPLAY / METER – This part uses the current or voltage
produced by the DELDIO,
detector andVeronica
amplifies it.Lourds A. device
The readout | BSP2Ethen
 Module 1: Exercise 1
Check-in
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United
7. DIGITAL DISPLAY / METER – This part uses the current or voltage produced by
the detector and amplifies it. The readout device then converts the current or
voltage into a form that the analyst can read as printout.

Module 1: Exercise 1
Post Task
Question 1
What is Beer’s Law?
The power of a transmitted radiant beam decreases exponentially as the
concentration of the solution containing the absorbing chemical species increases
arithmetically. The relationship can be expressed as A = εlc where A is absorbance,
ε is the molar extinction coefficient (which depends on the nature of the chemical
and the wavelength of the light used), l is the length of the path light must travel
in the solution in centimeters, and c is the concentration of a given solution.

Question 2
What are the characteristics of Beer’s Plot?
The plot of the data should be linear and should go through the origin If the plot
is not linear or if the y-intercept deviates substantially from the origin, it
indicates that the standards were improperly prepared, the samples deviate in
some way from Beer’s Law, or that there is an unknown interference in the sample

DELDIO, Veronica Lourds A. | BSP2E

that is complicating the measurements. Assuming a linear standard curve is
obtained, the equation that provides the best linear fit to the data is generated.
Calligraphers United
Beer's Law states that the concentration of a chemical solution is directly
proportional to its absorption of light. The premise is that a beam of light
becomes weaker as it passes through a chemical solution. The attenuation of light
occurs either because of distance through a solution or increasing concentration.
Under specific conditions, the Beer-Lambert law neglects to keep a straight
connection among weakening and centralization of the analyte. These deviations are
characterized into three classifications: (1) Genuine—essential deviations because
of the limits of the actual law. (2) Substance—deviations saw because of explicit
synthetic types of the example which are being broken down. (3) Instrument—
deviations that happen because of how the weakening estimations are made.

Question 3
What are the Applications of using UV/VIS Spectroscopy?
UV/VIS spectroscopy can be utilized in the assembling cycle to recognize impurities
inside a substance or measure the energy of a response. Distinguishing impurities,
many natural mixes will retain light in a specific locale of the UV range and foreign
substances can without much of a stretch be identified and evaluated with a
solitary estimation. The adjustment in absorbance at 284 nm is relative to the
measure of diazepam in an arrangement. Along these lines, basic and savvy
measures can be intended for the quantitative investigation of mixes.

DELDIO, Veronica Lourds A. | BSP2E

 Module 1: Exercise2
Pre-Task
Calligraphers United

Question 1
What is the goal of Infrared Spectroscopy?
Infrared Spectroscopy is the of infrared light interacting with a molecule. This can
be analyzed in three ways by measuring absorption, emission and reflection. The
main use of this technique is in organic and inorganic chemistry. It is used by
chemists to determine functional groups in molecules.

Question 2
What are the advantages of Fourier Transform Infrared Spectrophotometers?
An FT-IR instrument does not use a slit to limit the individual frequency reaching
the sample and detector as a dispersive instrument does. There are also fewer
mirror surfaces in an FT-IR spectrometer, so there are fewer reflection losses
than in a dispersive spectrometer.

DELDIO, Veronica Lourds A. | BSP2E

 Module 1: Exercise2
Check-In
Calligraphers Activity
United

Question 1
Draw and label the parts of IR Spectrophotometer?

Question 2
Discuss the function of the different parts of IR Spectrophotometer?
Radiation source - source of continuous infrared radiation and a sensitive
infrared transducer, or detector.
Sample cells and sampling of substances.
Monochromators - device used to disperse or separate a broad spectrum of
IR radiation into individual narrow IR frequencies
Detectors - reacts to infrared (IR) radiation. The two main types of
detectors are thermal and photonic (photodetectors)
Recorder - record IR spectrum

DELDIO, Veronica Lourds A. | BSP2E

 Module 1: Exercise2
Post Task
Calligraphers United

Question 1
Complete the table below:

Question 2
What are the solvent used to dissolve the sample in IR analysis?

Carbon disulfide
Carbon tetrachloride
Tetrachloroethylene
Chloroform
Dimethylformamide
Dioxane
Cyclohexane

DELDIO, Veronica Lourds A. | BSP2E

Question 3
Why does water not use as a solvent in IR Analysis?

Water cannot be used as a solvent for IR spectroscopy because

Water has two strong infrared absorption.
Water is a strongly polar solvent that dissolves the alkali halide
disks commonly used for IR.

DELDIO, Veronica Lourds A. | BSP2E