The Beer-Lambert Law

At the end of this topic, students should be able to:

Use Beer-Lambert’s Law to calculate the concentration of a given species in solution

Absorbance isn't very good for making comparisons

The importance of concentration

The proportion of the light absorbed will depend on how many molecules it interacts with. Suppose you
have got a strongly colored organic dye. If it is in a reasonably concentrated solution, it will have a very
high absorbance because there are lots of molecules to interact with the light.

However, in an incredibly dilute solution, it may be very difficult to see that it is colored at all. The
absorbance is going to be very low.

Suppose then that you wanted to compare this dye with a different compound. Unless you took care to
make allowance for the concentration, you couldn't make any sensible comparisons about which one
absorbed the lightest.

The importance of the container shape

Suppose this time that you had a very dilute solution of the dye in a cube-shaped container so that the
light travelled 1 cm through it. The absorbance isn't likely to be very high. On the other hand, suppose
you passed the light through a tube 100 cm long containing the same solution. More light would be
absorbed because it interacts with more molecules.

Again, if you want to draw sensible comparisons between solutions, you have to allow for the length of
the solution the light is passing through.

Both concentration and solution length are allowed for in the Beer-Lambert Law.

The Beer-Lambert Law

What the Law looks like ?

You should recognize the expression on the left of this equation as what we have just defined as the
absorbance, A. You might also find the equation written in terms of A:

That's obviously easier to remember than the first one, but you would still have to learn the equation for
absorbance. It might be useful to learn it in the form:

The Greek letter epsilon in these equations is called the molar absorptivity - or sometimes the molar
absorption coefficient.

Molar absorptivity

If you rearrange the simplest of the equations above to give an expression for epsilon (the molar
absorptivity), you get:

Remember that the absorbance of a solution will vary as the concentration or the size of the container
varies. Molar absorptivity compensates for this by dividing by both the concentration and the length of
the solution that the light passes through. Essentially, it works out a value for what the absorbance
would be under a standard set of conditions - the light travelling 1 cm through a solution of 1 mol dm-3.

That means that you can then make comparisons between one compound and another without having
to worry about the concentration or solution length.

Values for molar absorptivity can vary hugely. For example, ethanal has two absorption peaks in its UV-
visible spectrum - both in the ultra-violet. One of these corresponds to an electron being promoted from
a lone pair on the oxygen into a pi anti-bonding orbital; the other from a pi bonding orbital into a pi anti-
bonding orbital.

The table gives values for the molar absorptivity of a solution of ethanal in hexane. Notice that there are
no units given for absorptivity. That's quite common. If you want them, and assuming the length is in cm
and the concentration is mol dm-3, the units are mol-1 dm3 cm-1.
 electron jump wavelength of maximum absorption (nm) molar absorptivity

lone pair to pi anti-bonding orbital 290 15

pi bonding to pi anti-bonding orbital 180 10000

The ethanal obviously absorbs much more strongly at 180 nm than it does at 290 nm. (Although, in fact,
the 180 nm absorption peak is outside the range of most spectrometers.)

USING UV-VISIBLE ABSORPTION SPECTRA

Using UV-absorption spectra to help identify organic compounds

You should remember the Beer-Lambert Law:

The expression on the left of the equation is known as the absorbance of the solution and is measured
by a spectrometer. The equation is sometimes written in terms of that absorbance.

The symbol epsilon is the molar absorptivity of the solution.

Finding concentration using the molar absorptivity

If you know the molar absorptivity of a solution at a particular wavelength, and you measure the
absorbance of the solution at that wavelength, it is easy to calculate the concentration. The only other
variable in the expression above is the length of the solution. That's easy to measure and, in fact, the cell
containing the solution may well have been manufactured with a known length of 1 cm.

For example, let's suppose you have a solution in a cell of length 1 cm. You measure the absorbance of
the solution at a particular wavelength using a spectrometer. The value is 1.92. You find a value for
molar absorptivity in a table of 19400 for that wavelength.

Substituting those values:

Notice what a very low concentration can be measured provided you are working with a substance with
a very high molar absorptivity.

t is much better to measure the concentration by plotting a calibration curve. It saves doing any
calculations for one thing!

Finding concentration by plotting a calibration curve

Doing it this way you don't have to rely on a value of molar absorptivity, the reliability of the Beer-
Lambert Law, or even know the dimensions of the cell containing the solution.

What you do is make up a number of solutions of the compound you are investigating - each of
accurately known concentration. Those concentrations should bracket the concentration you are trying
to find - some less concentrated; some more concentrated. With coloured solutions, this isn't a problem.
You would just make up solutions of accurately known concentrations some of which are a bit lighter
and some a bit darker in colour.

For each solution, you measure the absorbance at the wavelength of strongest absorption - using the
same container for each one. Then you plot a graph of that absorbance against concentration. This is a
calibration curve.

According to the Beer-Lambert Law, absorbance is proportional to concentration, and so you would
expect a straight line. That is true as long as the solutions are dilute, but the Law breaks down for
solutions of higher concentration, and so you might get a curve under these circumstances.

As long as you are working from values either side of the one you are trying to find, that isn't a problem.
Having drawn a best fit line, the calibration curve will probably look something like the next diagram.
(I've drawn it as a straight line because it is easier for me to draw than a curve(!), and it's what you will
probably get if you are working with really dilute solutions. But if it turns out to be a curve, so be it!)

Notice that no attempt has been made to force the line back through the origin. If the Beer-Lambert Law
worked perfectly, it would pass through the origin, but you can't guarantee that it is working properly at
the concentrations you are using.

Now all you have to do is to measure the absorbance of the solution with the unknown concentration at
the same wavelength. If, for example, it had an absorbance of 0.600, you can just read the
corresponding concentration from the graph as below.