Spectrophotometry report

INTRODUCTION

A given chemical compound is made of atoms and bonds between those atoms. When

irradiated with electromagnetic radiation, the molecule and atoms can suffer electronic

transitions or other changes like bond vibrations due to given energy that has been absorbed. The

more substance there is, the more radiation is absorbed making the quantification of this

absorption a very useful property in several fields of chemistry. The technique based on the

quantification analysis of electromagnetic radiation is known as spectroscopy, and it bases its

principles in the Beer-Lambert Law which states that the relationship between the amount of

absorbed radiation (Absorbance) and the amount of substance (concentration) is linear. r in other

words, the more substance the more radiation is absorbed. This Law is expressed mathematically

as  A=ε ×c ×l

 Where  ε  is the molar absorption coefficient, c is the sample concentration and l the length of the

cell. In this lab session the concentration of an unknown sample of a CoCl2 solution, as well as a

coumarin solution, was determined. 

MATERIALS AND METHODS

Reagents Instruments
 Cobalt (II) chloride solutions
 Spectrophotometer
 Coumarin solutions
 Cuvettes
 Distilled water

1. Put a cuvette with distilled water in the spectrophotometer and set the Absorbance to 0.

Then remove the cuvette.

2. Place a 0.001 M solution of CoCl2 in the spectrophotometer, measure and record the

absorbance at different wavelengths from 350 nm to 700 nm.

 A
3. Calculate the value of molar absorption coefficient using the equation ε =
cl

4. Measure the absorbance of an unknown concentration CoCl2 solution at the maximum

recorded absorbance wavelength at step 2.

5. Determine the concentration of the unknown concentration solution

6. Repeat steps 2-5 for coumarin solutions.

DATA AND RESULTS

 CoCl2 solution.

A 0.1631 L
Absorbance Data for CoCl2 ε= = =163.1
Wavelength Absorbance cl mol mol ×cm
0.001 × 1 cm
(nm) (A) L
350 0.0348
400 0.049
450 0.1138 Calculations for unknown CoCl2 concentration:
500 0.1631
550 0.1088
A=6.4888 ( at 500 nm )
600 0.0259
650 0.0294
700 0.0351 A 6.4888 mol
c= = =0.040
εl L L
163.1 ×1 cm
Maximum wavelength = 500 nm mol × cm

c=0.001 M l=1 cm

 Coumarin solution.

Absorbance Data for Coumarin 550 0.0020

Wavelength Absorbance 600 0.0072
(nm) (A) 650 0.0153
350 -0.4405 700 -0.0023
400 0.2073 Maximum wavelength = 450 nm
450 0.2582
500 0.0024
 c=0.001 M l=1 cm A 12.0846 mol
c= = =0.048
A 0.2582 L εl L L
ε= = =253.8 253.8 ×1 cm
cl mol mol × cm mol × cm
0.001 × 1 cm
L
Unknown coumarin concentration:
A=12.0846 ( at 450 nm )

DISCUSSION

The wavelengths absorbed by compounds are different from each other because the chemical

bonds between the atoms that they are made of are different. This causes that the required energy

to produce electronic transitions are different and therefore the wavelength associated with the

radiation is also different. This is why the maximum absorbance wavelength needed to be

determine first besides from the fact that if another wavelength was chosen to determine the

concentration the linear correlation of the Beer-Lambert law could be lost, possibly leading to

incorrect concentration values of the unknowns.

For the CoCl2, the wavelength of maximum absorption was determined to be 500 nm, then the

molar extinction coefficient was determined (163.1 L/mol.cm) which is characteristic of the

compound. At this wavelgenht, the Beer-Lambert law is valid and therefore the concentration of

an unknown can be determined by measuring its absorbance at this particular wavelength. The

unknown concentration was determined to be 0.040 mol/L which was a lot higher than the

concentration of the known concentration solution.

Similarly the concentration of the coumarin solution with unknown concentration was

determined to be 0.048 M. The first difference between the two comes at the wavelength of

maximum absorption, since in the case of coumarin was 450 nm. This is due to the different kind
of compounds, the cobalt salt is an ionic compound while coumarin is an organic molecular

compound with different electronic configuration and therefore different absorption patterns.