Anthony F.

Desimone

U85516508

CHM2045L

Section 907

09/14/21

Post Lab #2

1) What is the relationship between absorbance and concentration?

Absorbance is directly proportional to concentration, therefore as concentration increases absorbance

increases and vice versa.

2) Which dilution technique is a more suitable choice for the purpose of creating a calibration curve?
How can you support your answer?

Parallel dilution is a more suitable choice as the calibration plot has a better linear fit than the
calibration plot formed through serial dilution. The R^2 value reflects how well the data fits the linear
regression line. The R^2 value for serial dilution was 0.8558 while the R^2 value for parallel dilution was
0.9999. A R^2 value closer to one means the data fits the linear regression line more. This is important in
creating a calibration curve as the linear regression line of the curve reflects beer’s law equation A=ebc.
The equation of the regression line has the form y=mx+b and in the context of a calibration curve y is
absorbance, x is concentration, m is the path length multiplied by the molar absorptivity coefficient (e x
b). Since the information provided from the linear regression line is needed for this investigation the
parallel dilution technique is the more suitable choice for this lab as it has the most linear fit.

3) If a solution is blue-colored, what wavelength of light (in nm) is being absorbed?

The wavelength of light in nm being absorbed of a blue-colored solution would be 600 nm as the
absorbed color would be orange and orange has an absorbed wavelength of 600 nm.

4) What is being measured in “absorption spectra”?

In an absorption spectra a sample’s absorbance at different wavelengths of UV-visible light is being

measured through a spectrometer. With the x-axis corresponding to the wavelength of light and the y-
axis corresponding to the absorbance. This spectrum provides the wavelength of maximum
absorbance(lmax).

5) Why is the wavelength of light set at maximum absorbance when making a Beer’s law plot?

Because at the wavelength of light of maximum absorbance the spectrometer will be more sensitive to
changes in absorbance which is important in creating a beer’s law plot as further apart points on the
plot allow for a more accurate linear regression line.
Table 1. Absorbance Values from serial dilution at 450 nm

Concentration Sample Volume of Volume of Concentration Absorbance

of Stock Stock Solution Water Added (M)
Solution (M) (mL) (mL)
A 1 9 2.5 x 10^-5 0.351
2.5 x 10^-4 B 1 9 2.5 x 10^-6 0.047
C 1 9 2.5 x 10^-7 0.02

Concentration Calculations:

Sample A:

M1V1=M2V2

(2.5 x 10^-4 M)(1 mL)=M2(10 mL)

M2=2.5 x 10^-5 M

Sample B:

M1V1=M2V2

(2.5 x 10^-5 M)(1 mL)=M2(10 mL)

M2=2.5 x 10^-6 M

Sample C:

M1V1=M2V2

(2.5 x 10^-6 M)(1 mL)=M2(10 mL)

M2=2.5 x 10^-7 M
Table 2. Absorbance Values from parallel dilution at 450 nm

Concentration Sample Volume of Volume of Concentration Absorbance

of Stock Stock Solution Water Added (M)
Solution (M) (mL) (mL)
A 1 9 2.50 x 10^-5 0.351
B 2 8 5.00 x 10^-5 0.665
2.5 x 10^-4 C 3 7 7.50 x 10^-5 0.903
D 4 6 1.00 x 10^-4 0.970
E 5 5 1.25x 10^-4 0.989
Concentration Calculations:

Sample A:

M1V1=M2V2

(2.5 x 10^-4 M)(1 mL)=M2(10 mL)

M2=2.5 x 10^-5 M

Sample B:

M1V1=M2V2

(2.5 x 10^-4 M)(2 mL)=M2(10 mL)

M2=5.00 x 10^-5 M
Sample C:

M1V1=M2V2

(2.5 x 10^-4 M)(3 mL)=M2(10 mL)

M2=7.5 x 10^-5 M

Sample D:

M1V1=M2V2

(2.5 x 10^-4 M)(4 mL)=M2(10 mL)

M2=1.0 x 10^-4 M

Sample E:

M1V1=M2V2

(2.5 x 10^-4 M)(5 mL)=M2(10 mL)

M2=1.25 x 10^-4 M
Concentration of Unknown Yellow dye Solution:

Table 3. Absorbance of Unknown Yellow dye Solution

Unknown Solution
Absorbance λmax (nm)
1.047 418.5

A=Absorbance

e=Molar Absorptivity Coefficient (L/mol-cm)

b=Path Length (cm)

C=Concentration (mol/L)

A=ebc ---> y=mx+b

m from the regression line collected from the parallel dilution plot is equal to (e x b), so

m= eb

13429= e(1 cm)

e=13429 (L/mol-cm)

Plugging e into beer’s law equation with the gathered data above in table 3 gives the concentration,

1.047=(13429 L/mol-cm)(1 cm)(c)

c= 7.8 x 10^-5 mol/L

So the concentration of the unknown yellow dye solution using beers law and the parallel dilution
calibration plot is 7.8 x 10^-5 mol/L.