CHIU, Ina Cathrina R. SALACUP, Esther Eunice U.

SPECTROSCOPY
the science in which the interactions of radiation and matter are used for qualitative and quantitative analysis of matter

SPECTROPHOTOMETRY
a subset of spectroscopy which involves a procedure that uses light interaction with matter in order to measure the concentration of a substance SPECTROPHOTOMETER Apparatus that measure the amount of light absorbed at a given wavelength

TRANSMITTANCE
Attenuation
Particles of the solution absorb electromagnetic radiation. Energy is transferred into the molecules of the solution. Intensity of radiation transmitted out of the solution is decreased.

Absorbing solution of concentration c Po P

Because of the interaction between photons and the particles of the solution, the beam decreases from Po to P.

TRANSMITTANCE
the fraction of incident radiation transmitted by the solution Often expressed as percent transmittance

T = Transmittance Po = transmitted intensity of light P = incident transmission

ABSORBANCE
measure of how much radiant power is absorbed by the solution

Inversely related to transmittance absorbance = transmittance Directly related to concentration of absorbing species concentration = absorbance

BEERS LAW
(absorption law)

A = absorbance = molar absorptivity (M-1 cm-1) b = path length c = concentration of absorbing species in M

Beers Law shows that there is a linear relationship between absorbance and concentration of absorbing species.

Beers Law
For mixtures, the total absorbance is the sum of the individual absorbance

Atotal = 1bc1 + 2bc2 + + nbcn

Methodology
A. Determination of Concentration Range for Analysis of KMnO4 in solution

Prepare 100 mL of 0.001M KMnO4 and 0.001M K2Cr2O7 solutions.

In 20 mL test tubes, prepare (10) 10 mL solutions by diluting 0.001M KMnO4 solution with distilled water in the following ratio: 0:10, 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, 9:1

Warm up the spectrophotometer for 20 minutes. Adjust the wavelength to 545 nm.

Insert the cuvette with the reference blank (distilled water) and set the absorbance (A) and transmittance (T) to 100%.

Read the absorbance of the prepared solutions, starting from the least concentrated.

Measure the absorbance of the unknown solution.

B. Spectrophotometric Determination of Mn and Cr in a Mixture Prepare a 2:8 dilution of K2Cr2O7 (Procedure A) and read the A of the soln at 545 nm and 440 nm.

A 2:8 dilution of KMnO4 soln at 440 nm

Calculate the concentration of MnO4-1 and Cr2O7-2 in the mixture as ppm Mn and ppm Cr, respectively.

Measure the A of the unknown soln at 545 nm and 440 nm.

Results and Discussion

Determination of Concentration Range for Maximum Precision
Dilution 0:10 1:9 2:8 3:7 4:6 5:5 6:4 7:3 2:8 Concentration [MnO42-] 0 1 x 10-4 M 2 x 10-4 M 3 x 10-4 M 4 x 10-4 M 5 x 10-4 M 6 x 10-4 M 7 x 10-4 M 8 x 10-4 M ppm Mn 0 5.494 10.99 16.48 21.98 27.47 32.96 38.46 43.95 Absorbance 545 nm 0 0.165 0.442 0.668 0.914 1.175 1.425 1.651 1.877 440 nm 0 0.010 0.028 0.048 0.061 0.073 0.089 0.094 0.126 %Transmittance 545 nm 100% 68.4% 36.1% 21.5% 12.2% 6.68% 3.76% 2.23% 1.33% 440 nm 100% 97.7% 93.8% 89.5% 86.9% 84.5% 81.5% 80.5% 74.8%

1:9

9 x 10-4 M

49.95

2.130

0.140

0.741%

72.4%

Sample computations for 1:9 dilution of KMnO4

Concentration range for max precision Correlation Coefficient, r Absorbance of unknown Concentration of unknown (ppm Mn)

0.9996 0.537 13.14 ppm

Linear regression - plot A (545 nm) vs. ppm Mn m = b = 0.04382 b = -0.038875 X = concentration of Mn in ppm

= 13.14 ppm

Absorbance (545 nm) vs. ppm Mn

2.5 2

Absorbance

1.5
1 0.5 0 0 -0.5 10 20 30 40 50 60

ppm Mn

%Transmittance (545 nm) vs. ppm Mn

120% 100%

% Transmittance

80%
60% 40%

20%
0% 0 10 20 30 40 50 60

ppm Mn

Results and Discussion

Dilution 0:10 1:9 2:8 3:7 4:6 5:5 6:4 Concentration [Cr2O72-] 0 1 x 10-4 M 2 x 10-4 M 3 x 10-4 M 4 x 10-4 M 5 x 10-4 M 6 x 10-4 M ppm Cr 0 10.349 20.798 31.198 41.597 51.996 62.395 0.002 0.003 0.007 Absorbance 545 nm 0 440 nm 0 0.048 0.095 0.142 0.185 0.230 0.268 99.5% 99.3% 98.4% %Transmittance 545 nm 100% 440 nm 100% 89.5% 80.4% 72.1% 65.3% 58.9% 54.0%

7:3 2:8
1:9

7 x 10-4 M 8 x 10-4 M
9 x 10-4 M

72.794 83.194
93.593 0.013

0.311 0.354
0.392 97.1%

48.9% 44.3%
40.6%

Concentration range for max precision Correlation Coefficient, r

0.9995

Absorbance of unknown Concentration of unknown (ppm Mn)

0.193 44.52 ppm

Linear regression -plot A (440 nm) vs. ppm Cr m = b = 4.1804 x 10-3 b = 6.8731 x 10-3 X = concentration of Mn in ppm
= 44.52 ppm

Absorbance (440 nm) vs. ppm Cr

0.45 0.4

0.35
0.3

Absorbance

0.25

0.2
0.15 0.1 0.05 0 0 20 40 60 80 100

ppm Cr

%Transmittance (440 nm) vs. ppm Cr

120%

100%

% Transmittance

80% 60% 40% 20% 0%

20

40

60

80

100

ppm Mn

Spectrophotometric Determination of Mn and Cr in a Mixture

KMnO4 = Max = 545 nm K2Cr2O7 = Max = 440 nm
K2Cr2O7 KMnO4

Beers Law can be applied to solutions containing more than one kind of absorbing substance provided that there is no interaction among the various species.

A 1 = 1 x bcx + 1 y bcy (at 545 nm) A 2 = 2 x bcx + 2 y bcy (at 440 nm)
Pagkatapos ng nakakaiyak na pagiisolate...

LEGEND: 1 = A at 545 nm 2 = A at 440 nm x = Mn y = Cr

Unknown Mn (x) A at 545 nm (1) A at 440 nm (2) 0.537 0.193 Cr (y)

Known Mn + Cr 1.4335 0.146

Concentration of Mn (ppm) : 32.6630 ppm Concentration of Cr (ppm) : 12.3949 ppm

Calculating for (b)Mn,545 , (b)Cr,545 , (b)Mn,440, and (b)Cr,440 :

Linear regression (plot A at 545 vs ppm Mn) (b)Mn,545 = slope = 0.043827 ppm-1 cm-1 Linear regression (plot A at 440 vs ppm Mn) (b)Mn,440 = slope = 2.7986 x 10-3 ppm-1 cm-1 Linear regression (plot A at 545 vs ppm Cr) (b)Cr,545 = slope = 1.4466 x 10-4 ppm-1 cm-1

Linear regression (plot A at 440 vs ppm Cr) (b)Cr,440 = slope = 4.1803 x 10-3 ppm-1 cm-1

Concentration of Mn (ppm)

CMn = 32.663 ppm

Concentration of Cr (ppm)

CCr = 12.3949 ppm

Answers to Guide Questions

How does absorbance vary with concentration? Absorbance is directly proportional to concentration. If the plot of A against C is extrapolated, will it intersect the point of origin?
Yes, it will only intersect the point of origin when there is no

absorbing species present in the sample. This was 0:10 solution where in there was 0 ml of KMnO4 and 10 ml distilled water, which may be considered as a reference blank.

How does % Transmittance vary with concentration? Transmission refers to how much light remains after it passes through. As the absorber concentration goes up, the absorbance goes up, but the transmission and the percent transmittance go down. In short, they have an indirect relationship.

Answers to Guide Questions

How does curve of A vs. C compare with that obtained by plotting %T vs. C? Explain the difference.
2.5 2 1.5 A(Y) 1 40 0.5 0 0 10 20 30 C 40 50 60 20 0 0 10 20 30 C 40 50 60
%T
A

120 100 80 60 %T

Answers to Guide Questions

There is a linear relationship between absorbance and concentration in dilute solutions. A plot of absorbance vs. concentration gives a straight line at a particular wavelength and temperature. On the other hand, percent transmittance is indirectly proportional to the concentration, resulting in a highly non-linear plot, an exponential plot in fact.

Answers to Guide Questions

A 20.0 ppm solution of a colored compound gives a 70.0% transmittance in a 1.00cm cell. Find the absorptivity of the solution. A = abc *a for absorptivity, for molar absorptivity* A = -log T

Solution:

A = -log 0.7 = 0.1549 0.1549 = a (1.0 cm) (20.0 ppm) a = 7.745 x 10-3

Answers to Guide Questions

A solution of a colored species gives 25% transmittance. What will be the % transmittance of the solution whose concentration is three times that of the given? Assume linearity of response over the given concentration range. A = bc A = -log T %T = (T/To) x 100 Solution: %T = 25% T = 0.25 A = -log 0.25 A = 0.602
A = b (3c) -> 3A [bec. of linearity] 3A = 3(0.602) = 1.806 (A at 3x concentration) 10-1.806 = 0.0156 = T

%T = T x 100 = 1.56%

Final Answer!

25 % to 1.56% WOW! Anlaki ng pagbaba!

Recommendations
It is important to remember not to have fingerprints on the cuvette because fingerprints absorb and scatter light slightly, even though they might not be readily visible.
The samples containing the standard solutions could be photodecomposed, therefore, the vessels that contain the standard solution must be amber bottles.

Recommendations
When the solutions are placed in test tubes, as much as possible, the sample must be exposed to minimal or totally no light in order for the sample not to absorb the light which could contribute to the reflection and scattering losses, which are major error contributors to the experiment proper.