http://uncw.edu/chem/Courses/Reeves/OnLineLabs/scienceMajors/BeersLaw_PH.

pdf, 05/12/2014

9s
Beers Law
OBJECTIVES

To observe the connection between the concentration of a chromophore (colored substance), the length
of the sample that the light passed through (path length), and the intensity of the solutions color.

To perform dilutions and relate concentrations obtained for diluted materials to the concentrations of
the undiluted products.

To determine the concentration of food dye in a commercial product using Beers Law.

INTRODUCTION
Light can be thought of as energy traveling in waves through space. These waves can be described in terms
of wavelength (, distance between peaks or troughs of a wave, c.f. Figure 9.1) or frequency (, number of
peaks or troughs passing a particular point in space per unit time).

Figure 9.1
Humans visually perceive a very narrow range of wavelengths, called visible light, between 750 and 400
nm (Remember: 1 nm = 10-9 m). This visible spectrum encompasses all of the colors that we can see, and,
as can be seen in Figure 9.2 below, each color has a particular wavelength region associated with it.

Figure 9.2
A substance will appear a particular color because it absorbs visible light corresponding to its
complimentary color, which is the color that is directly opposite absorbed light on the color wheel (Figure
9.3). This may seem a bit counterintuitive because what you are observing is what is not being absorbed by
the solution. Therefore a blue dye appears blue because it absorbs light of wavelengths that do not
correspond to blue light (i.e. orange etc.) and transmits blue light.

Figure 9.3
The absorption of light by a substance in a solution can be described mathematically by the Beer-Lambert
Law:
A = ebc
where:
A = absorption at a given wavelength of light,
e = molar absorptivity, unique to each molecule and varying with wavelength,
b = the path length through the solution that the light has to travel, and
c = the concentration of the solution in moles per liter (molarity).
Lets look at these terms individually. The more light that is absorbed by a solution, the more deeply
colored the solution is in the region of the spectrum that it is not absorbing. So if a solution is deeply
colored, we would expect it to have a large absorbance (A) in the wavelength region where it absorbs. That
the absorbance (A) would increase with increasing concentration (c) should make intuitive sense: a more
concentrated solution absorbs more light because it has more molecules, atoms or ions in it (for a given
volume of liquid) to absorb. One drop of food coloring in 5 mL of water will be less deeply colored, and
have a corresponding lower value of A, than 4 drops of food coloring in 5 mL of water. The absorbance (A)
also increases with increasing path length. This is also somewhat intuitive. If the light passes through 10 cm
of the solution, more light will be absorbed (larger A, more deeply colored) than if the light only passes
through 1 cm of solution. You can prove this using a dilute solution of food coloring in water. If you place
the food coloring in a large skinny container, like a salt shaker, the solution will appear more deeply

colored if you look up through the salt shaker than if you look at it through the side (see the Figure 9.4
below):

Figure 9.4
The molar absorptivity is unique to whatever the absorbing species is. The larger e is, the larger the
corresponding A will be. Some molecules have enormous molar absorptivities in certain regions of the
visible spectrum; these are often used as dyes because a small amount absorbs a great deal of light and thus
the substance will appear very deeply colored (in the wavelength region where the dye doesnt absorb).
Even if two substances absorb in the same wavelength region and have the same concentration, they may
appear very different in terms of the depth of color, due to differences in their molar absorptivities.
If the absorption of a particular chromophore is measured in a series of solutions in which its concentration
is varied systematically, a "Beer's Law Plot" (Figure 9.5) can be generated. This plot can act as a
"calibration curve" because it can be used to determine the concentration of the chromophore in an
unknown solution by measuring the solution's absorbance (A). Note that a proportional relationship exists
between the absorbance and the concentration of the chromophore.

Figure 9.5
Once the calibration curve is drawn, the concentration of an unknown sample may be determined by
measuring the absorption of the sample at the same wavelength and pathlength.

Colorimetry.
Colorimetry is based on a particular application of Beers Law, focusing upon the relationship between
absorption and path length. If you compare two solutions of the same light absorbing species, the molar
absorptivities (e) will be identical. Thus any combination of path length and concentration that give the
same product (b*c) will absorb the same amount of light. Then, if two salt shakers containing solutions
with different concentrations of a particular chromophore appear to have the same depth of color when
viewed from above, their absorption will be the same:
A 1 = A2
Since A = ebc, and e1 = e2 (since it is the same substance), we can write:
b1c1 = b2c2
This relationship is useful in determining an unknown concentration by comparison to a known
concentration. Mathematically:
bknowncknown = bunknowncunknown

In this experiment, you will use solutions of food coloring of known concentrations to determine the
concentrations of food coloring in one or two consumer products. You will combine two colorimetric
analysis techniques, the standard series method and the balancing method. In the standard series method,
the color intensity (which is the eyes way of determining absorption) of the unknown solution is compared
with the color intensities of a series of standard solutions (solutions of accurately known
concentration). The color comparison is made in nearly identical vessels (salt shakers), and solutions are
compared by viewing through the lengths of the columns (i.e. vertically). The unknown is found to have an
intensity somewhere between two of the successive standard solutions, thus establishing its concentration
to be within a certain range. The actual concentration of the unknown (cunknown) is then determined using the
balancing method, which matches the color intensities more closely. In the balancing method you remove
small amounts of the unknown solution until its intensity matches that of the known solution. When the two
solutions appear to have the same color intensity, we can assume
Aknown = Aunknown
bknowncknown = bunknown
Thus, by measuring the heights of the unknown and standard solutions (bunknown and bknown), one can
determine the concentration of the unknown:

cunknown =

bknown cknown
bunknown

EXPERIMENTAL

blue food coloring (Blue #1)

yellow food coloring (Yellow #6)
small cups or bottles (100 mL or
greater)
measuring spoons and cups
2 (minimum, 6 work best) salt
shakers or other long, narrow vessels.
commercial products that contain
only Blue #1 or Yellow #6
dye. (Examples: Fla-Vor-Ice pops,
various blue mouthwashs, Gatorade.)

Standard Solutions
Below are results for the determination of the concentration of commercially available food coloring that
you buy in the small dropper bottles at the supermarket.
Dye

Molar Absorptivity (M-1cm-1)

Concentration of Commercial Food Coloring (M)

Blue #1

1.3x10

0.026

Yellow # 5

3.6x104

0.112
Procedure

Preparation of standard dye solutions from commercial (grocery store) food coloring solutions.
(The commercial food coloring solutions are quite concentrated. You will need to dilute them to make a
series of standard solutions of known concentrations. )
For each of the food dyes:
1.

Measure out 1/8 teaspoon (tsp) of the commercial food coloring and dilute to 1 quart (qt). We will
call this Solution A.

2.

You will use Solution A to start a series of serial dilutions to prepare your standard solutions. Use
the charts below. (Note: Store the standard solutions in your small cups. Be sure to label the
cups.)
Blue # 1

Volume of
Solution Diluting Solution
Diluting Solution
A
concentrate (0.026 M) 1/8 tsp (0.62 ml)
B
A
1 Tsp (15 mL)
C
A
1 tsp (5 mL)
D
B
1 tsp (5 mL)
E
C
1 tsp (5 mL)

Volume of
Water (mL)
1 qt (946 mL)
1 Tsp (15 mL)
3 Tsp (45 mL)
3 Tsp (45 mL)
3 Tsp (45 mL)

Final
total Volume Concentration (M)
1 qt (947 mL)
1.7x10-5
2 Tsp (30 mL)
8.5x10-6
3.3 Tsp (50 mL)
1.7x10-6
3.3 Tsp (50 mL)
8.5x10-7
3.3 Tsp (50 mL)
1.7x10-7

Yellow #6
Volume of
Diluting Solution
Diluting Solution
concentrate (0.112 M) 1/8 tsp (0.62 ml)
A
1 Tsp (15 mL)
A
1 tsp (5 mL)
B
1 tsp (5 mL)
C
1 tsp (5 mL)

Volume of
Water (mL)
1 qt (946 mL)
1 Tsp (15 mL)
3 Tsp (45 mL)
3 Tsp (45 mL)
3 Tsp (45 mL)

Final
total Volume Concentration (M)
1 qt (947 mL)
7.3x10-5
2 Tsp (30 mL)
3.7x10-5
3.3 Tsp (50 mL)
7.3x10-6
3.3 Tsp (50 mL)
3.7x10-6
3.3 Tsp (50 mL)
7.3x10-7

Preparation of unknown samples

Dilute the commercial product that contains the food coloring by a factor of 1:10. To do this, take 2 tsp (10
ml) of the product and dilute with 6 TBS (90ml) of water. NOTE: You may ultimately need to dilute this
solution further if it doesnt match up properly with any of the standard solutions.
Determination of food dye concentration in unknown sample.
Colorimetric Procedure
(This procedure works best if you have six salt shakers, one for each standard solution and one for your
unknown. However you can do the procedure if you have only two salt shakers; it simply becomes a bit
more cumbersome. The instructions below assume only two salt shakers, but if you have six, you can simply
compare solution intensities without needing to do all the transferring. )
1.

Transfer the unknown solution to the marked clean salt shaker. Transfer standard solution E to its
corresponding clean, marked salt shaker. Using a sheet of white paper placed flat upon a surface such
as a table or kitchen counter as a background, hold the salt shaker containing unknown solution next to
the standard E salt shaker. Look directly down both to compare color intensities.

2.

Replace the solution in the standard salt shaker with each of the other standards, comparing color
intensities each time, until you find the one that matches most closely with the unknown solution. You
should find that the color intensity of your unknown solution lies between that of two successive
standards, thus establishing a concentration range for the unknown.

3.

If not, either dilute standard E (if E is more intense than the unknown sample) or the unknown sample
(if the sample is more intense than intense than standard A). Do this by performing similar dilutions to
those from the tables above. Be sure to keep track of the dilutions you make so you can determine the
final concentration of the standard and the dye.

4.

When you have found the two successive standard solutions whose intensities span that of the
unknown, select the standard of lower concentration to continue with the balancing method.
Balancing method.

5.

Using the standard solution selected above, place it side by side with your unknown solution against
the white background. If you have selected an appropriate standard, you will find that its color is
slightly less intense than that of the unknown solution.

6.

Carefully use a dropper to remove small volumes of the unknown solution from the salt shaker until
the color intensities of the unknown and standard in the salt shakers are matched very closely.

7.

Measure the heights of the columns of standard and unknown solutions, using a ruler, and record your
data in the appropriate space in your table.

Repeat this entire process for as many samples of commercial products containing dye that you were able
to obtain.

RESULTS & DISCUSSION

Complete the following:
Submit your completed Data Sheet to your instructor.
Write a laboratory report that details your observations about the experiment. Be sure show all calculations
associated with determining the concentraiton of dye in the commercial products. Submit your report along
with your completed Data Sheet to your instructor.

DATA SHEET
Name of
Product

Identity of matching
standard (A - E)

Mstd solution

Depth of standard
solution

Depth of
Unknown

Munknown

Morig product