ABSTRACT

The purpose of performing this instrument called UV/Visible Spectrophotometry is to

determine the wavelength at the maximum absorbance. Besides that, it is a study about visible
absorption spectrum and how to handle Ultraviolet Visible Spectrophotometer accurately also
practice students to use the Beer`s Law. In order to determine the maximum absorbance, Potassium
Manganate (KMnO4) is diluted originally from concentration of 1000 ppm to 100ppm at first and
later from the 100ppm it is diluted once again to 10, 20, 40 and 50 ppm. All samples are filled up
until reach 2/3 cell. Then, the sample is put into Spectrophotometer one by one. The maximum
absorbance got are 0.310 for 20 ppm and 0.769 for 50 ppm while the wavelength got from these
two maximum absorbance is the same, that is 525 nm. The Beer`s Law is used to calculate
absorptivity,A and transmittance,T. The absorptivities got are 15.6 for 20 ppm and 16.0

for 50 ppm. The transmittance got are 0.4875 for 20 ppm and 0.1578 for 50 ppm. The
concentration of both unknowns are determined which are X are 36.565 ppm and for unknown
solution Y is 23.482 ppm. From the experiment, all values for absorbance got are in positive value.
Apparently from the spectra plotted manually, it shows that the spectrum is band spectra.

OBJECTIVE

i. To obtained a visible absorption spectrum of KmnO4 plotted manually by recorder.

ii. To obtained the wavelength of maximum absorbance from the spectrum, and absorptivity
calculated from Beer's Law.
iii. The determination of unknown concentraion of a sample by standard curve method.
PRINCIPLE OF INSTRUMENT

Many chemical species absorb light in either the visible or ultra violet region of the
electromagnetic spectrum. This absorption is caused by the changing electron energy levels in the
molecule and is therefore characteristic of the molecular structure. The technique is particularly
useful for performing rapid quantitative concentration measurements of organic species in solution.
Many molecules absorb ultraviolet or visible light. The absorbance of a solution increases as
attenuation of the beam increases. Absorbance is directly proportional to the path length, b, and the
concentration, c, of the absorbing species. Beer's Law states that :
A = abc, where a is a constant of proportionality, called the absorbtivity.

A = Absorbance (no unit, since A = )

a = Absorptivity ( )
b = Path length of the sample (cm)
c = Concentration of the compound in solution ( )

OR
A = bc

A = Absorbance (no unit, since A = )

a = Absorptivity ( )
b = Path length of the sample (cm)
= Concentration of the compound in solution ( )

A solution whose solute concentration is 1 ppm contains 1 g of solute for each million (106)
grams of solution or equivalently, 1 mg of solute per kg of solution. Because the density of water is
1 g/ml, 1 kg of a dilute aqueous solution will have a volume very close to 1 l. Thus,

1 ppm = 1 mg of solute per liter of solution

Part per million (ppm) = 106

Different molecules absorb radiation of different wavelengths. An absorption spectrum will show a
number of absorption bands corresponding to structural groups within the molecule.
CHEMICAL & APPARATUS

• standard solution KmnO4

• DI water

• beaker
• volumetric flask
• dropper
• pipette
• tissue
• gloves

METHODOLOGY

A. Preparation of Potassium Manganate (KMnO4) with concentration of 100 ppm.

1. The volume of Potassium Manganate (KMnO4) needed is calculated before the dilution
process is started (The calculation is shown in calculation part).
2. 25 ml of Potassium Manganate (KMnO4) of 1000 ppm is pipetted and transferred into 250
ml volumetric flask.
3. The distilled water is added into volumetric flask until the red line level is achieved.
4. The volumetric flask is shaken to make sure that Potassium Manganate (KMnO4) is well
distributed.

B. Preparation of Potassium Manganate (KMnO4) with concentration of 10 ppm.

1. The volume of Potassium Manganate (KMnO4) needed is calculated before the dilution
process is started (The calculation is shown in calculation part).
2. 5 ml of Potassium Manganate (KMnO4) of 100 ppm is pipetted and transferred into 50 ml
volumetric flask.
3. The distilled water is added into volumetric flask until the red line level is achieved.
4. The volumetric flask is shaken to make sure that Potassium Manganate (KMnO4) is well
distributed.

C. Preparation of Potassium Manganate (KMnO4) with concentration of 20 ppm.

1. The volume of Potassium Manganate (KMnO4) needed is calculated before the dilution
 process is started (The calculation is shown in calculation part).
2. 10 ml of Potassium Manganate (KMnO4) of 100 ppm is pipetted and transferred into 50 ml
volumetric flask.
3. The distilled water is added into volumetric flask until the red line level is achieved.
4. The volumetric flask is shaken to make sure that Potassium Manganate (KMnO4) is well
distributed.

D. Preparation of Potassium Manganate (KMnO4) with concentration of 40 ppm.

1. The volume of Potassium Manganate (KMnO4) needed is calculated before the dilution
process is started (The calculation is shown in calculation part).
2. 20 ml of Potassium Manganate (KMnO4) of 100 ppm is pipetted and transferred into 50 ml
volumetric flask.
3. The distilled water is added into volumetric flask until the red line level is achieved.
4. The volumetric flask is shaken to make sure that Potassium Manganate (KMnO4) is well
distributed.

E. Preparation of Potassium Manganate (KMnO4) with concentration of 50 ppm.

1. The volume of Potassium Manganate (KMnO4) needed is calculated before the dilution
process is started (The calculation is shown in calculation part).
2. 25 ml of Potassium Manganate (KMnO4) of 100 ppm is pipetted and transferred into 50 ml
volumetric flask.
3. The distilled water is added into volumetric flask until the red line level is achieved.
4. The volumetric flask is shaken to make sure that Potassium Manganate (KMnO4) is well
distributed.

F. Determination of absorption of Potassium Manganate (KMnO4) using Ultraviolet Visible

Spectrometer.
1. The cuvettes are cleaned using soft tissue paper.
2. Two cuvettes are filled up with distilled water until reach of the cell.
3. Both cuvettes are placed into the Ultraviolet Visible Spectrophotometer and the absorption
value is measured.
4. The absorption value is recorded in the table 1.
5. One of the cuvette is taken out and rinsed with distilled water.
6. The distilled water that still attached at the cuvette is wiped out using soft tissue paper.
7. The cuvette is filled up with Potassium Manganate (KMnO4) of 10 ppm until reach of the
 cell.
8. The cuvette is then placed into the Ultraviolet Visible Spectrophotometer and the absorption
value is measured.
9. The absorption value is recorded in the table 1.
10. Step 5-9 are repeated for Potassium Manganate (KMnO4) of 20 ppm, 40 ppm, 50 ppm.
Note: All steps in handling the spectrophotometer is referred to the laboratory assistant.

CALCULATION
A. Calculation of dilution :
i) Dilution of stock solution, Potassium Manganate (KMnO4) from 1000 ppm to 100 ppm
=
(1000 ppm) = (100 ppm)(10 ml)
= 1 ml

ii) Dilution of Potassium Manganate (KMnO4) from 100 ppm to 10 ppm

=
(100 ppm) = (10 ppm)(10 ml)
= 1 ml

iii) Dilution of Potassium Manganate (KMnO4) from 100 ppm to 20 ppm

=
(100 ppm) = (20 ppm)(10 ml)
= 2 ml

iv) Dilution of Potassium Manganate (KMnO4) from 100 ppm to 40 ppm

=
(100 ppm) = (40 ppm)(10 ml)
= 4 ml

v) Dilution of Potassium Manganate (KMnO4) from 100 ppm to 50 ppm

=
(100 ppm) = (50 ppm)(10 ml)
= 5 ml
B. Calculation of absorptivity :
i. for 20ppm
1 ppm = 1 mg of solute per liter of solution

Thus,
20 ppm = 20 mg of solute per liter of solution

= 0.02

From Beer`s Law,

A = abc

a=

a = 15.6

ii. for 50ppm

50 ppm = 50 mg of solute per liter of solution

= 0.05

From Beer`s Law,

A = abc

a=

a = 16.0
C. Calculation of transmittance
i. for 20ppm
A=2-
=2–A
%T=

%T=

T=

T = 0.4875
ii. for 50ppm
A=2-
=2–A
%T=

%T=

T=

T = 0.1578
DATA & RESULT

Wavelength, (nm) Blank ^(20 ppm) ^(50 ppm)

700 0.000 0.008 0.015
650 0.000 0.026 0.049
600 0.000 0.032 0.074
550 0.000 0.275 0.692
540 0.000 0.271 0.683
530 0.000 0.290 0.724
525 0.000 0.310 0.769
520 0.000 0.272 0.683
500 0.000 0.191 0.475
450 0.000 0.030 0.065
400 0.000 0.037 0.061
350 0.000 0.187 0.412
Table 1 : measurement of A and for 20ppm & 50ppm.

Concentration (ppm) Absorbance

10 0.1502
20 0.3150
40 0.6251
50 0.7881
X 0.5740
Y 0.3660
Table 2 : fixed wavelength absorbance measurement.

DISCUSSION
CONCLUSION

From this experiment, the concentration of both unknowns are determined which are X
are 36.565ppm and for unknown solution Y is 23.482ppm. This experiment able the students on
handling the UV-Vis Spectrophotometry and also the functions. In this experiment, the Beer-
Lambert Law is useful for characterizing many compounds but does not hold as a universal
relationship for the concentration and absorption of all substances. By using this law the values of
transmittance can be calculated. Eventually, this experiment is successfully achieved its aim.

REFFERENCES

Modern Instrumental Analysis by Satinder Ahuja, Neil D. Jespersen, Volume 47 (2006) 864
pages.

Principle of Instrumental Analysis by Douglas A. Skoog, F. James Holler, Stanley R.

Crouch, Sixth Edition (2007) 1039 pages.

http://www2.chemistry.msu.edu:80/faculty/reusch/VirtTxtJml/Spectrpy/UV-Vis/uvspec.htm