Chemistry 182

Erin M
Thursdays 7:30
Experiment 1: Visible Spectroscopy, Atomic and Molecular

Objective: Beers law, which correlates absorbance with concentration of a solution, and Lambert's law, which
correlates absorption with path length of a solution, allows for the calculating the molar absorptivity if the path
length is known. By measuring the different absorptions for different concentrations of solution, and graphing
that data, well be able to calculate the concentration of two different unknowns. This is a observation method
used in many types of experiments that allows for the calculation of concentration based off of the absorbance.
Methods: The concentration of the five test tubes is determined through the relation of molarity times volume
of stock solution will equal the molarity times the volume of the diluted solution. Using a Spectrophotometer we
are able to shine light through the solution and determine the amount of light absorbed by a solution which
allows us to quantized the color of a solution and calculate the concentration. The amount of light that is
absorbed is based off the chemical structure and electrons in a molecule or compound, the wavelength of the
light shined on the solution and the concentration of the solution. The spectrophotometer will give us a reading
of the %T which is the transmittance and a measurement of how strongly the solution absorbs the light shone
on it. With the %T we can calculate the absorbance units of the solution at that wavelength. This unit is
proportional to a few other constants which is where Beers Law will be used. Beers Law states that
absorbance is equal to absorptivity ( a constant unique to each compound or molecule ) times the path length,
times the concentration. Plotting the measured absorbance rates of different concentrations against the
concentration values in a graph will give us an equation for Beers Law, that can be used to calculate the
concentrations of unknowns solutions.
Data:
Table 1: Preparation of the Working Standard
Concentration of Stock Alphazurine (M)

1.454E-04

Volume of Stock used (mL)

10.00

Volume of Working Sol'n Made (mL)

100.0

Concentration of Working Sol'n (M)

1.500E-05

Table 2: Preparation of Calibration Curve Standards

Lrg.TT#

Vol. Reading for Working

(mL)

Vol. Reading for Water

(mL)

Vol. of Working

Vol. of Water

Initial

Final

Initial

Final

Used (mL)

Used (mL)

2.17

3.19

0.10

7.01

1.02

6.91

1.29

3.31

0.02

6.10

2.02

6.08

0.11

3.22

0.00

5.29

3.11

5.29

0.11

4.10

0.10

4.28

3.99

4.18

0.00

4.99

0.01

3.10

4.99

3.09

Table 3: Spectral Analysis of Alphazurine Samples at 628 nm using Spec 20 #24

Sm.TT #

%T (run 1)

%T (run 2)

%T (run 3)

%T (avg)

Conc. (M)

Blank

100.0

100.0

99.8

99.9

0.00E+00

0.000

73.4

73.0

73.2

73.2

1.93E-06

0.135

56.8

56.8

57.2

56.9

3.74E-06

0.245

45.8

45.8

45.8

45.8

5.55E-06

0.339

36.6

36.8

36.8

36.7

7.33E-06

0.435

28.2

28.0

27.8

28.0

9.26E-06

0.553

Working

14.4

14.6

14.6

14.5

1.50E-05

0.838

Stock

2.4

2.2

2.2

2.3

1.45E-04

1.645

Unk # 45

40.2

38.6

40.2

See Table 5

Unk # 26

58.0

58.4

58.0

See Table 5

Table 4: Beer's Law Plot for Analysis of Unknowns

Slope of Beer's Law Plot (Blank, 1-5)

5.84E+04

Intercept of Beer's Law Plot (Blnk, 1-5)

1.38E-02

Table 5: Report Table for Alphazurine Unknowns

Unk #

Run

Absorbance

Conc (M)

Accepted

% Error

High/Low

45

0.396

6.54E-06

6.3678E-06

2.71

high

0.413

6.84E-06

7.46

high

0.396

6.54E-06

2.71

high

Avg

0.402

6.64E-06

4.30

high

S.D.

0.010

0.237

3.81E-06

1.40

low

0.234

3.76E-06

2.72

low

0.237

3.81E-06

1.40

low

Avg

0.236

3.80E-06

1.84

low

S.D.

0.002

26

3.869E-06

Calculations:
1. Concentration of working solution
((stock solution molarity )*(stock solution volume))/(Volume of working solution)
((1.454E-04)*(.0100 L )/(.10000 L )
=1.500E-05 M
2. Volume of working standard per test tube
( Final volume ) - ( initial volume )
Excel =(C12-B12)
=1.02 mL
3. Concentration of each test tube
(molarity of working solution * vol of working solution )/( vol of working soln + vol of water )
Excel = ($D$7*F12)/(F12+G12)
=1.93E-06 M
4. Average % T
Excel = AVERAGE(B20:D20)
=%73.2
5. Absorbance
Excel = LOG10(100/E21)
=0.135
6. Slope and Intercept
Excel Chart
Slope 5.84E05 M-1 cm-1
Intercept 1.38E-02 M
7. Unknown Concentration
(absorbance - intercept)/(slope)
Excel =(C42-$D$33)/$D$32
= 6.64E-06 M
8. Percent Error
((accepted value- expt value)/(expt value))*100
=((D42-$E$39)/$E$39)*100
=4.30 % high

Report Table:
Experimental Value

Accepted Value

Percent Error

high/low

Unknown
Solution #45

6.64E-06 M

6.3678E-06 M

4.30%

high

Unknown
Solution #26

3.80E-06 M

3.8689E-06 M

1.84%

low

Discussion Questions:

1. Using the linear plot of absorbance vs concentration from the first graph, the molar absorptivity (e) for the
colored solution can e calculated using 10.0 mm as the path length. That would equal to the slope = the molar
absorptivity times the path length. So 10*e=5.840E4 equals out to the molar absorptivity equaling 5840.
2. It matters if the large test tubes are dry but not the volumetric flask because the volumetric flask is used to
make the diluted solution so if there will need to be a little bit of water in it anyways, you may want to make
sure there isnt too much water so that it wont go over the line when you add the stock solution though. The test
tubes have to be dry because you dont want to add any more water into the solution which would dilute it
slightly and could effect your calculations.
3. If you diluted the stock solution in a 100 mL volumetric flask and under-filled the flask this would cause your
determination of the concentration of the unknown be calculated as lower then it actually is. When you made
the graph the concentration number would be calculated as lower then it actually is, which would result in the
%T being higher then it would be at the calculated concentration. That wouldn't change the slope of the line but
when you would look at the absorbance of the unknown on the line, it would put you at a lower concentration
for that absorbance.
4. The first graph was used to calculate the concentrations of the unknown instead of the second graph because
the linear relationship between the absorbance and concentrations starts to plateau at high concentrations so its
more of a curve with a straight line towards higher values of concentration. Putting a linear equation onto these
values will give you an inaccurate slope and intercept which will make your calculations of the concentration
not be accurate. As long as you are calculation a concentration that is within the linear portion of the relation of
absorbance and concentration, then you can use the slope of that line to calculate it.
5. If you were given an Unknown Solution that had an Absorbance above the linear range of your Beers Law
plot, to best determine the concentration you could start the plot at a point a little bit before and slightly after
your absorbance amount and use the linear equation from that data to calculate the concentration. If your
absorbance is really high you will calculate a really high concentration. The accuracy will not be as close but its
a good general estimation.