Experiment 1: Spectrophotometry

Determination of Concentration by Light

Absorbance

Course: Chemistry 103.1 Spring 2016, Tuesday 9:40AM-12:30PM

Name: Elizabeth Joa
Partners Name: Roderick Pena
Date Experiment was Performed: 2/16/16
Date Report is Due: 2/23/16

Purpose
In this lab experiment, an unknown of potassium dichromate solution was given to find
out what the concentration of the solution was. To do this, we analyze the absorbance
measurements while using a standard curve and also Beer-Lambert law.
Introduction
The human eye sees color because substances absorb visible light and also reflect light.
The colors reflected are the colors we see with our eye. Light is a form of energy in the
form of waves. A spectrophotometer is an instrument used to measure light absorbance.
The absorbance of 0 indicates that that particular light was not absorbed. The
concentration of the solution determines the amount of light that is absorbed or goes
through the solution. The solution tested in this instrument absorbs specific wavelength
of light. From the data we collect using a spectrophotometer, we can use the BeerLambert law to determine our concentration. In this experiment, we will also create a
standard curve to help us determine the unknown concentration of the solution, in
addition to calculating with Beer-Lambert law. When we plot the absorbance of the
known solution concentrations, we can determine the concentration of the unknown with
the standard curve created.

Figure 1: A sample in a spectrophotometer measuring the light absorbance. Some light

was absorbed and some that went through the solution.
Beer-Lambert Law:
A=cl
A: absorbance
: extinction coefficient
c: concentration
l: path length of the sample holder

Method
The first step of this experiment was to prepare the samples of different concentrations of
potassium dichromate in 6 different test tubes. Once the samples were made, with each
sample, we added some of the sample in the colorimeter vials. But before measuring their
absorbance, we calibrated the spectrophotometer to set to zero absorbance. Once we
added the vials into the spectrophotometer, we looked at the colorimeter display to
obtain the values of the Blue (468 nm) absorbance. We recorded the absorbance of all the
6 samples.
1
Water (mL) 10.0
potassium
0
dichromate
, 2.0 mM
(mL)
Unknown
potassium
dichromate
(mL)

2
8.0
2.0

3
6.0
4.0

4
4.0
6.0

5
2.0
8.0

6
0
-

10.0

Table 1: Potassium dichromate solution and water amounts used to prepare the samples.
In order to be able to find the standard curve, we need to determine the concentration of
potassium dichromate of each sample, using MAVA=MBVB
After receiving all the calculations of the concentration and collecting the data for the
absorbance, a standard curve was created and we were able to determine the
concentration of the unknown potassium dichromate sample from the absorbance
measurement.
Results
Results #1
Volume, mL
0
2.0
4.0
6.0
8.0
10.0

Concentration, mM
0
0.40
0.80
1.2
1.6
Unknown= 1.25

Absorbance
0
0.033
0.284
0.421
0.604
0.456

Discussion
In conclusion, the spectrophotometer is an accurate instrument used to measure the
concentration of an unknown solution. The spectrophotometer is also accurate in
measuring the absorbance of the solutions. After creating the linear regression analysis,
the values plotted on the standard curve gave a line of best fit that gave a reading of the
unknown potassium dichromate concentration as 1.25 M. The line of best fit is very close
to the plotted concentration/absorbance values of test tubes 1-5. When calculating the
concentration of the unknown with the Beer-Lambert law, the answer came out to 1.56
M. The answer from the calculation with the Beer-Lambert law and also from the linear
regression analysis are very close in value with only a .31 difference. Therefore, my
results for finding the value of the concentration of the unknown potassium dichromate
are accurate and exceptionally successful.
Post- Lab Discussion Questions for Experiment #1
1. Beers Law states that as the concentration of a substance in a solution becomes
greater, its absorbance will increase.
2. The purpose of using a reagent blank in the experiment is to calibrate the
instrument. It sets the absorbance to zero and blanks the colorimeter.
3. The standard line must be a best fit that passes through the origin because it is
more accurate that the data began at zero absorbance and zero concentration.
4. A) If a solution appears to be purple, yellow color of light is absorbed by the
solution.
B) Violet/Purple
5. We construct a standard curve that relates concentration to absorbance because
with this standard line, we can find any concentration or absorbance of any
unknown solution. We also want to measure the accuracy of the instrument we
used and if its calibrated.

Reference
Chemistry 103.1 Lab Manual (Spring 2016)