Absorption spectroscopy

Absorption spectroscopy refers to spectroscopic techniques that measure the absorption

of radiation, as a function of frequency or wavelength, due to its interaction with a
sample. The sample absorbs energy, i.e., photons, from the radiating field. The intensity
of the absorption varies as a function of frequency, and this variation is the absorption
spectrum. Absorption spectroscopy is performed across the electromagnetic spectrum.

Absorption spectroscopy is employed as an analytical chemistry tool to determine the

presence of a particular substance in a sample and, in many cases, to quantify the amount
of the substance present. Infrared and ultraviolet-visible spectroscopy are particularly
common in analytical applications. Absorption spectroscopy is also employed in studies
of molecular and atomic physics, astronomical spectroscopy and remote sensing.

There are a wide range of experimental approaches to measuring absorption spectra. The
most common arrangement is to direct a generated beam of radiation at a sample and
detect the intensity of the radiation that passes through it. The transmitted energy can be
used to calculate the absorption. The source, sample arrangement and detection technique
vary significantly depending on the frequency range and the purpose of the experiment.

Basic Theory
More technically,[1][2] absorption spectroscopy is based on the absorption of photons by
one or more substances present in a sample, which can be a solid, liquid, or gas, and
subsequent promotion of electron(s) from one energy level to another in that substance.
Note that the sample can be a pure, homogeneous substance or a complex mixture. The
frequency at which the incident photon is absorbed is determined by the difference in the
available energy levels of the different substances present in the sample; it is the
selectivity of absorbance spectroscopy - the ability to generate photon (light) sources that
are absorbed by only some of the components in a sample - that gives absorbance
spectroscopy much of its utility. Typically, X-rays are used to reveal chemical
composition, and near ultraviolet to near infrared wavelengths are used to distinguish the
configurations of various isomers in detail. In absorption spectroscopy the absorbed
photons are not re-emitted (as in fluorescence) rather, the energy that is transferred to the
chemical compound upon absorbance of a photon is lost by non-radiative means, such as
transfer of energy as heat to other molecules.

While the relative intensity of the absorption lines do not vary with concentration, at any
given frequency the measured absorbance ( − log(I / I0)) has been shown to be
proportional to the molar concentration of the absorbing species and the thickness of the
sample the light passes through. This is known as the Beer-Lambert law. The plot of
amount of radiation absorbed versus frequency for a particular compound is referred to as
the absorption spectrum. The normalized absorption spectrum is characteristic for a
particular compound, does not change with varying concentration and is like the chemical
"fingerprint" of the compound. At frequencies corresponding to the resonant energy
levels of the sample, some of the incident photons are absorbed, resulting in a drop in the
measured transmission intensity and a corresponding dip in the spectrum. The absorption
spectrum can be measured using a spectrometer and by knowing the shape of the
spectrum ,the optical path length and the amount of radiation absorbed, one can
determine the structure and concentration of the compound.

Relation To Emission Spectroscopy

Emission is a process by which a substance releases energy in the form of

electromagnetic radiation. Emission can occur at any frequency at which absorption can
occur, and this allows the absorption lines to be determined from an emission spectrum.
The emission spectrum will typically have a quite different intensity pattern from the
absorption spectrum, though, so the two are not equivalent. The absorption spectrum can
be calculated from the emission spectrum using appropriate theoretical models and
additional information about the quantum mechanical states of the substance.

Relation To Scattering and Reflection Spectroscopy

The scattering and reflection spectra of a material are influenced by both its index of
refraction and its absorption spectrum. In an optical context, the absorption spectrum is
typically quantified by the extinction coefficient, and the extinction and index
coefficients are quantitatively related through the Kramers-Kronig relation. Therefore,
the absorption spectrum can be derived from a scattering or reflection spectrum. This
typically requires simplifying assumptions or models, and so the derived absorption
spectrum is an approximation.

Applications
Analytical Chemistry

Absorption spectroscopy is useful in chemical analysis because of its specificity and its
quantitative nature. The specificity of absorption spectra allows compounds to be
distinguished from one another in a mixture. For example, absorption spectroscopy is
used to identify the presence of pollutants in the air, distinguishing the pollutant from the
nitrogen, oxygen, water and the other expected constituents.[3] The specificity also allows
unknown samples to be identified by comparing a measured spectrum with a library of
reference spectra. In many cases, it is possible to determine qualitative information about
a sample even if it is not in a library. Infrared spectra, for instance, have characteristics
absorption bands that indicate if carbon-hydrogen or carbon-oxygen bonds are present.

An absorption spectrum can be quantitatively related to the amount of material present

using the Beer-Lambert law. Determining the absolute concentration of a compound
requires knowledge of the compound's absorption coefficient. The absorption coefficient
for some compounds is available from reference sources, and it can also be determined
by measuring the spectrum of a calibration standard with a known concentration of the
target.
Circular dichroism
First pioneered by Jean-Baptiste Biot, Augustin Fresnel, and Aimé Cotton [1], circular
dichroism (CD) refers to the differential absorption of left and right circularly polarized
light.[2][3]. This phenomenon is exhibited in the absorption bands of optically active chiral
molecules. CD spectroscopy has a wide range of applications in many different fields.
Most notably, UV CD is used to investigate the secondary structure of proteins[4]. UV/Vis
CD is used to investigate charge-transfer transitions [5]. Near-infrared CD is used to
investigate geometric and electronic structure by probing metal d→d transitions[6].
Vibrational circular dichroism, which uses light from the infrared energy region, is used
for structural studies of small organic molecules, and most recently proteins and DNA[7].

Physical Principles
Circular polarization of light

Main article: Circular polarization

Electromagnetic radiation consists of an electric and magnetic field that oscillate

perpendicular to one another and to the propagating direction [8]. While linearly polarized
light occurs when the electric field vector oscillates only in one plane and changes in
magnitude, circularly polarized light occurs when the electric field vector rotates about its
propagation direction and retains constant magnitude. Hence, it forms a helix in space
while propagating. For left circularly polarized light (LCP) with propagation towards the
observer, the electric vector rotates counterclockwise[9]. For right circularly polarized
light (RCP), the electric vector rotates clockwise.
Optical rotatory dispersion (ORD)
Optical rotatory dispersion is the variation in the optical rotation of a substance with a
change in the wavelength of light. Optical rotatory dispersion can be used to find the
absolute configuration of metal complexes. For example, when plane-polarized white
light from an overhead projector is passed through a cylinder of sucrose solution, a spiral
rainbow is observed perpendicular to the cylinder.[citation needed]

When white light passes through a polarizer, the extent of rotation of light depends on its
wavelength. Short wavelengths are rotated more than longer wavelengths. Because the
wavelength of light determines its color, the variation of color with distance through the
tube is observed.[citation needed] This dependence of specific rotation on wavelength is called
optical rotatory dispersion.

Infrared spectroscopy
Infrared spectroscopy (IR spectroscopy) is the subset of spectroscopy that deals with
the infrared region of the electromagnetic spectrum. It covers a range of techniques, the
most common being a form of absorption spectroscopy. As with all spectroscopic
techniques, it can be used to identify compounds and investigate sample composition. A
common laboratory instrument that uses this technique is an infrared spectrophotometer.

The infrared portion of the electromagnetic spectrum is usually divided into three
regions; the near-, mid- and far- infrared, named for their relation to the visible spectrum.
The far-infrared, approximately 400–10 cm−1 (1000–30 μm), lying adjacent to the
microwave region, has low energy and may be used for rotational spectroscopy. The mid-
infrared, approximately 4000–400 cm−1 (30–2.5 μm) may be used to study the
fundamental vibrations and associated rotational-vibrational structure. The higher energy
near-IR, approximately 14000–4000 cm−1 (2.5–0.8 μm) can excite overtone or harmonic
vibrations. The names and classifications of these subregions are merely conventions.
They are neither strict divisions nor based on exact molecular or electromagnetic
properties.

Theory
Infrared spectroscopy exploits the fact that molecules absorb specific frequencies that are
characteristic of their structure. These absorptions are resonant frequencies, i.e. the
frequency of the absorbed radiation matches the frequency of the bond or group that
vibrates. The energies are determined by the shape of the molecular potential energy
surfaces, the masses of the atoms, and the associated vibronic coupling.

In particular, in the Born–Oppenheimer and harmonic approximations, i.e. when the

molecular Hamiltonian corresponding to the electronic ground state can be approximated
by a harmonic oscillator in the neighborhood of the equilibrium molecular geometry, the
resonant frequencies are determined by the normal modes corresponding to the molecular
electronic ground state potential energy surface. Nevertheless, the resonant frequencies
can be in a first approach related to the strength of the bond, and the mass of the atoms at
either end of it. Thus, the frequency of the vibrations can be associated with a particular
bond type.

[edit] Number of vibrational modes

In order for a vibrational mode in a molecule to be "IR active," it must be associated with
changes in the permanent dipole.

A molecule can vibrate in many ways, and each way is called a vibrational mode. Linear
molecules have 3N-5 degrees of vibrational modes whereas nonlinear molecules have
3N-6 degrees of vibrational modes (also called vibrational degrees of freedom). As an
example H2O, a non-linear molecule, will have 3*3-6 = 3 degrees of vibrational freedom,
or modes.

Simple diatomic molecules have only one bond and only one vibrational band. If the
molecule is symmetrical, e.g. N2, the band is not observed in the IR spectrum, but only in
the Raman spectrum. Unsymmetrical diatomic molecules, e.g. CO, absorb in the IR
spectrum. More complex molecules have many bonds, and their vibrational spectra are
correspondingly more complex, i.e. big molecules have many peaks in their IR spectra.