Aspirin 

(USAN), also known as acetylsalicylic acid (pronounced /əˌsɛtəlˌsælɨˈsɪlɨk/ ə-SET-əl-sal-i-SIL-ik,

abbreviated ASA), is a salicylate drug, often used as an analgesic to relieve minor aches and pains, as
an antipyretic to reduce fever, and as an anti-inflammatory medication.

Salicylic acid, the main metabolite of aspirin is integral part of human and animal metabolism. While much
of it is attributable to diet a substantial part is synthesized endogenously.[1]

Aspirin also has an antiplatelet effect by inhibiting the production of thromboxane, which under normal
circumstances binds platelet molecules together to create a patch over damaged walls of blood vessels.
Because the platelet patch can become too large and also block blood flow, locally and downstream,
aspirin is also used long-term, at low doses, to help prevent heart attacks, strokes, and blood
clot formation in people at high risk for developing blood clots.[2] It has also been established that low
doses of aspirin may be given immediately after a heart attack to reduce the risk of another heart attack
or of the death of cardiac tissue.[3][4]

The main undesirable side effects of aspirin are gastrointestinal ulcers, stomach bleeding, and tinnitus,

especially in higher doses. In children and adolescents, aspirin is no longer used to control flu-like
symptoms or the symptoms of chickenpox or other viral illnesses, because of the risk of Reye's
syndrome.[5]

Aspirin was the first discovered member of the class of drugs known as nonsteroidal anti-inflammatory
drugs (NSAIDs), not all of which are salicylates, although they all have similar effects and most have
inhibition of the enzyme cyclooxygenase as their mechanism of action. Today, aspirin is one of the most
widely used medications in the world, with an estimated 40,000 tonnes of it being consumed each year.
[6]
 In countries where Aspirin is a registeredtrademark owned by Bayer, the generic term is acetylsalicylic
acid (ASA).[7][8]

A spectrophotometer is employed to measure the amount of light that a

sample absorbs. The instrument operates by passing a beam of light through a
sample and measuring the intensity of light reaching a detector.

The beam of light consists of a stream of photons, represented by

the purple balls in the simulation shown below. When a photon encounters an
analyte molecule (the analyte is the molecule being studied), there is a chance
the analyte will absorb the photon. This absorption reduces the number of
photons in the beam of light, thereby reducing the intensity of the light beam.

You can visualize this process by running the simulation shown below. Click
on the Start button to start the simulation and the Stop button to stop the
simulation.

The light source is set to emit 10 photons per second. Watch the motion of the
photons and observe how some of the photons are absorbed (removed) as the
beam of light passes through the cell containing the sample solution. The
intensity of the light reaching the detector is less than the intensity emitted by
the light source.

The following simulation illustrates the procedures for making

spectrophotometric measurements.

 First, the intensity of light (I0) passing through a blank is measured.

The intensity is the number of photons per second. The blank is a
solution that is identical to the sample solution except that the blank
does not contain the solute that absorbs light. This measurement is
necessary, because the cell itself scatters some of the light.

 Second, the intensity of light (I) passing through the sample solution
is measured. (In practice, instruments measure the power rather than
the intensity of the light. The power is the energy per second, which is
the product of the intensity (photons per second) and the energy per
photon.)

 Third, the experimental data is used to calculate two quantities:

the transmittance (T) and the absorbance (A).

I
T =
I0
A = - log10 T

The transmittance is simply the fraction of light in the original beam that
passes through the sample and reaches the detector. The remainder of the light,
1 - T, is the fraction of the light absorbed by the sample. (Do not confuse the
transmittance with the temperature, which often is given the symbol T.)
 In most applications, one wishes to relate the amount of light absorbed to the
concentration of the absorbing molecule. It turns out that the absorbance rather
than the transmittance is most useful for this purpose. If no light is absorbed,
the absorbance is zero (100% transmittance). Each unit in absorbance
corresponds with an order of magnitude in the fraction of light transmitted.
For A = 1, 10% of the light is transmitted (T = 0.10) and 90% is absorbed by
the sample. For A = 2, 1% of the light is transmitted and 99% is absorbed.
For A = 3, 0.1% of the light is transmitted and 99.9% is absorbed.

Using the simulation below, perform the following steps:

 Measure the intensity of light passing through the blank.

 Measure the intensity of light passing through the sample.
 Calculate the transmittance.
 Calculate the absorbance.

Note: For each measurement, run the simulation long enough to detect at least
1000 photons. There is substantial random error in the intensity, and the more
photons that are counted, the lower the relative uncertainty in the results.

Iron(III) chloride, also called ferric chloride, is an industrial scale commodity chemical compound, with
the formula FeCl3. The colour of iron(III) chloride crystals depends on the viewing angle: by reflected light
the crystals appear dark green, but by transmitted light they appear purple-red. Anhydrous iron(III)
chloride is deliquescent, forming hydrated hydrogen chloride mists in moist air. It is rarely observed in its
natural form, mineralmolysite, known mainly from some fumaroles.

When dissolved in water, iron(III) chloride undergoes hydrolysis and gives off heat in

an exothermic reaction. The resulting brown, acidic, andcorrosive solution is used as
a coagulant in sewage treatment and drinking water production, and as an etchant for copper-based
metals inprinted circuit boards. Anhydrous iron(III) chloride is a fairly strong Lewis acid, and it is used as a
catalyst in organic synthesis.

THE BEER-LAMBERT LAW

This page takes a brief look at the Beer-Lambert Law and explains
the use of the terms absorbance and molar absorptivity relating to
UV-visible absorption spectrometry.

Absorbance

Measuring the absorbance of a solution

If you have read the page about how an absorption spectrometer

works, you will know that it passes a whole series of wavelengths
of light through a solution of a substance (the sample cell) and also
through an identical container (the reference cell) which only has
solvent in it.

Note:  It isn't essential to read about how

the spectrometerworks, but you could follow this link if you
are interested or if it is on your syllabus. Everything you
need from that page to understand the present topic is
repeated below.

For each wavelength of light passing through the spectrometer, the

intensity of the light passing through the reference cell is
measured. This is usually referred to as Io - that's I for Intensity.

The intensity of the light passing through the sample cell is also
measured for that wavelength - given the symbol, I.

If I is less than Io, then obviously the sample has absorbed some of
the light. A simple bit of maths is then done in the computer to
convert this into something called the absorbance of the sample -
given the symbol, A.

For reasons to do with the form of the Beer-Lambert Law (below),

the relationship between A (the absorbance) and the two intensities
is given by:

On most of the diagrams you will come across, the absorbance

ranges from 0 to 1, but it can go higher than that.

An absorbance of 0 at some wavelength means that no light of that

particular wavelength has been absorbed. The intensities of the
sample and reference beam are both the same, so the ratio Io/I is 1.
Log10 of 1 is zero.

An absorbance of 1 happens when 90% of the light at that

wavelength has been absorbed - which means that the intensity is
10% of what it would otherwise be.

In that case, Io/I is 100/I0 (=10) and log10 of 10 is 1.

Note:  If you don't feel comfortable with logarithms, don't

worry about it. Just accept that an absorbance scale often
runs from zero to 1, but could go higher than that in
extreme cases (in other words where more than 90% of a
wavelength of light is absorbed).

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
coloured 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 coloured 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 most light.

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 will find that various different symbols are given for some of
the terms in the equation - particularly for the concentration and the
solution length. I'm going to use the obvious form where the
concentration of the solution is "c" and the length is "l".

Note:  That's obviously "l" for length. The font I'm using
won't distinguish between "l" for length and a capital letter
"I" (for Intensity). That problem disappears in the equation
below - where it is obvious which is which.

You should recognise 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.
 Note:  These jumps are described in detail on the page
explaining the theory of UV-visible spectrometry.
Depending on your background knowledge, you may have
to read another page first, but there is a link to that on the
theory page.

Use the BACK button on your browser to return to this

page later if you choose to follow this link.

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.

wavelength of
molar
electron jump maximum absorption
absorptivity
(nm)
lone pair to pi anti-
290 15
bonding orbital
pi bonding to pi
180 10000
anti-bonding orbital

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.)

You may come across diagrams of absorption spectra plotting

absorptivity on the vertical axis rather than absorbance. However, if
you look at the figures above and the scales that are going to be
involved, you aren't really going to be able to spot the absorption at
290 nm. It will be a tiny little peak compared to the one at 180 nm.

To get around this, you may also come across diagrams in which
the vertical axis is plotted as log10(molar absorptivity).

If you take the logs of the two numbers in the table, 15 becomes
1.18, while 10000 becomes 4. That makes it possible to plot both
values easily, but produces strangely squashed-looking spectra!
Beer-Lambert Law, more commonly known as Beer's Law, states that the optical absorbance of a
chromophore in a transparent solvent varies linearly with both the sample cell pathlength and the
chromophore concentration. Beer's Law is the simple solution to the more general description of
Maxwell's far-field equations describing the interaction of light with matter. In practice, Beer's Law is
accurate enough for a range of chromophores, solvents and concentrations, and is a widely used
relationship in quantitative spectroscopy.

Absorbance is measured in a spectrophotometer by passing a collimated beam of light at wavelength λ

through a plane parallel slab of material that is normal to the beam. For liquids, the sample is held in an
optically flat, transparent container called a cuvette. Absorbance (Aλ) is calculated from the ratio of light
energy passing through the sample (I0) to the energy that is incident on the sample (I):

Aλ = -log (I/I0)

Beer's Law follows:

Aλ = ελbc

ελ = molar absorptivity or extinction coefficient of the chromophore at wavelength λ (the

optical density of a 1-cm thick sample of a 1 M solution). ελ is a property of the material and
the solvent.

b = sample pathlength in centimeters

c = concentration of the compound in the sample, in molarity (mol L-1)

In an absorbance experiment, light is attenuated not only by the chromophore, but also by reflections from
the interface between air and the sample, the sample and the cuvette, and absorbance by the solvent.
These factors can be quantified separately, but are often removed by defining I0 as the light passing
through a sample "blank" or "baseline" or reference sample (for example, a cuvette filled with solvent but
zero concentration of the chromophore is used as the blank).

Many factors can affect the validity of Beer's Law. It is usual to check for the linearity of Beer's Law for a
chromophore by measuring the absorbance of a series of standards. This "calibration" can also remove
errors in the experiment, the equipment, and the batch of reagents (such as cuvettes of unknown
pathlength).