LAB REPORT
BEER LAMBERT’S
LAW EXPERIMENT
DETERMING THE CONCENTRATION OF UNKNOWN ASA
SOLUTIONS
Objectives
We will learn:
About the absorption of light by molecules
Basic components of a spectrophotometer
How to prepare standards diluting a stock solution
How to Prepare a Beer’s Law curve from standard solutions
To Use a Beer’s Law curve to calculate the concentration of an unknown substance
Gain experience pipetting
Gain experience weighing small samples
Safety Notes
Eye protection must be worn at all times
Sodium hydroxide is caustic and should not come in contact with your skin or clothing.
Wear gloves when handling this chemical. A lab coat or lab apron is recommended.
Theory
Sample ASA Acetylsalicylic acid
Acetylsalicylic acid, commonly known as aspirin, absorbs light in the UV region of the electromagnetic
spectrum. The Spectronic device 200 operates in the visible region. Therefore, we must perform a series
of chemical reactions to convert acetylsalicylic acid to a colored complex, as shown in Figure 5. In
reaction 1, a base (e.g., sodium hydroxide) hydrolyzes acetylsalicylic acid to yield salicylate dianion. In
reaction 2, acidification converts the dianion to a monoanion, which complexes with iron (III) in reaction
3 to produce a violet-colored complex.
Beer lambert’s law
The primary objective of this experiment is to determine the concentration of an unknown
Acetylsalicylic acid solution. The Acetylsalicylic acid solution used in this experiment has a blue color, so
Colorimeter users will be instructed to use the red LED. A higher concentration of the colored solution
absorbs more light (and transmits less) than a solution of lower concentration.
You will prepare five of known concentration (standard solutions). Each solution is transferred to a
small, rectangular cuvette that is placed into the Colorimeter or Spectrometer. The amount of light
penetrates the solution and strikes the photocell is used to compute the absorbance of each solution.
When you graph absorbance vs. concentration for the standard solutions, a direct relationship should
result. The direct relationship between absorbance and concentration for a solution is known as Beer’s
law.
We will determine the concentration of an unknown Acetylsalicylic acid solution by measuring its
absorbance. By locating the absorbance of the unknown on the vertical axis of the graph, the
corresponding concentration can be found on the horizontal axis. The concentration of the unknown can
a so be found using the slope of the Beer’s law curve.
Spectrophotometers
Spectrophotometers are used to measure the amount of light absorbed or transmitted by a sample. The
instrument disperses white light into its component wavelengths by passing light through either a prism
or a grating. The intensity of light at any wavelength can be measured with a detector, such as a
photocell, a photomultiplier tube, or a solid-state device called a charge-coupled device (CCD). The
optical path of the Spectronic 200 used in today’s experiment is shown below in Figure:
Procedure:
Part 1: Prepare the beer’s law plot:
1. To make the beer’s plot we will prepare series of standard solution of ASA and measure their
absorbance at 530 nm.
2. Weigh 0.1g of ASA and record mass on data sheet.
3. Transfer to 50 ml Erlenmeyer flask.
4. Add 5ml of 1 M NaOH.
5. Heat to the boiling on the hot plate.
6. Transfer it to the volumetric flask and dilute it to the neck mark.
7. Mix it well by putting the top on the flash and invert the flash. This is now called stock solutions.
8. Transfer 0.1 ml / 0.2 ml/ 0.3 ml / 0.4 ml / 0.5 ml of stock solution to the volumetric flask with
FeCl3.KCl.HCl solution to the mark and label them as standard solution A, B, C, D, E, F
respectively.
9. Measure %transmittance at 530 nm and record it on data sheet
Part 2: analyze on aspirin tablet
1. To make the beer’s plot we will prepare series of standard solution of ASA and measure their
absorbance at 530 nm.
2. Weigh 0.1g of ASA and record mass on data sheet.
3. Transfer to 50 ml Erlenmeyer flask.
4. Add 5ml of 1 M NaOH.
5. Transfer it to the 50ml volumetric flask and dilute it to the neck mark.
6. Mix it well by putting the top on the flash and invert the flash. This is now called stock solutions.
7. Transfer 0.50ml to 10ml flask and dilute with FeCl3.KCl.HCl
8. Measure the %transmittance and record on data collection.
Calculations and Observations:
For this example, we assume 0.400 g of acetylsalicylic acid (aspirin, C9H8O4) is treated as outlined in the
procedure (you should use the actual mass recorded on your report sheet). The concentration of
complex in the stock solution can be found as follows:
Molar mass of ASA = 180.2 g/mol
Concentration of ASA is = mass of ASA used / Molar mass of ASA
Molarity of complex standard solution= (Vol. of stock sol.) (molarity of stock sol.) / total volume
This stock solution is used to prepare standards for the Beer’s Law curve by diluting aliquots of the stock
solution into 10.00 mL volumetric flasks, and diluting to volume with iron (III) chloride solution. The
concentration of C9H8O4 in the standards is calculated with the relationship M1V1 = M2V2, where M1 is
the concentration of the stock solution, V1 is the volume of the stock solution transferred, V2 is the
volume of the diluted solution (10.00 mL for all standards in this experiment), and M2 is the new
concentration.
Part 1:
Agenda 1st Determination 2nd Determination
Mass of aspirin sample 0.101 0.11
% Transmittance at 530nm of 10ml solution 91.8 92.9
Absorbance at 530nm of 10ml solution 0.037 0.032
Concentration of ASA in 10ml solution 0.000565934 0.000587912
Concentration of ASA in 50ml solution 0.00282967 0.00293956
Moles of ASA in 50ml solution 0.005659341 0.005879121
Mass of ASA in 50ml solution 0.101 0.11
Mass % of ASA in sample 44.2 45.6
Average Mass % of ASA in sample 44.9 46.88
Total mass of aspirin containing 250mg of ASA 325mg 331mg
Part 2:
Standard Mass of ASA Molarity of % Absorbance
log(%T)
Solution Used (g) solution (M) Transmittance 2-LOG(%T)
A 0.108 0.00059 71.6 1.855 0.145
B 0.158 0.00087 47.6 1.678 0.322
C 0.208 0.00114 34.8 1.542 0.458
D 0.258 0.00142 21.4 1.330 0.680
E 0.308 0.00169 16.6 1.220 0.780
Concentration (M) vs. Absorbance (A)
0.900
0.780
0.800
f(x) = 588.52 x − 0.2
0.700 R² = 0.99 0.670
0.600
Absorbance (A)
0.500 0.458
0.400
0.322
0.300
0.200 0.145
0.100
0.000
0.00040 0.00060 0.00080 0.00100 0.00120 0.00140 0.00160 0.00180
Concentration (M)
Conclusion:
By plotting the curve for ASA solution of different concentrations, we can now find the concentration of
any unknown solution by measuring the %transmittance or absorbance via spectrophotometer and
comparing with the curve as plotted.
