What is Spectroscopy?

the study of the interaction between matter and

electromagnetic radiation.
the interaction of ultraviolet, visible, and infrared radiation with
matter. Because these techniques use optical materials to
disperse and focus the radiation, they often are identified as
optical spectroscopies.
Electromagnetic Radiation

is a form of energy whose behavior is described by the

properties of both waves and particles. Some properties of
electromagnetic radiation, such as its refraction when it passes
from one medium to another, are explained best by describing
light as a wave.
Wave properties of Electromagnetic Radiation

Electromagnetic radiation consists of oscillating electric and

magnetic fields that propagate through space along a linear
path and with a constant velocity.
The oscillations in the electric and magnetic fields are
perpendicular to each other, and to the direction of the waves
propagation.
 an example of plane-
polarized electromagnetic
radiation, consisting of a
single oscillating electric
field and a single oscillating
magnetic field.

Plane-polarized electromagnetic radiation showing the oscillating electric field in red

and the oscillating magnetic field in blue. The radiations amplitude, A, and its
wavelength, , are shown. Normally, electromagnetic radiation is unpolarized, with
oscillating electric and magnetic fields present in all possible planes perpendicular to the
direction of propagation.
INTERACTION OF ELECTROMAGNETIC RADIATION WITH
MATTER
The interaction of radiation with matter can cause redirection
of the radiation and/or transitions between the energy levels of
the atoms or molecules.
PARTICLE PROPERTIES OF ELECTROMAGNETIC
RADIATION
When matter absorbs electromagnetic radiation it undergoes a
change in energy. The interaction between matter and
electromagnetic radiation is easiest to understand if we assume
that radiation consists of a beam of energetic particles called
photons.
 Photons
is absorbed by a sample it is destroyed, and its energy
acquired by the sample.3 The energy of a photon, in joules,
is related to its frequency, wavelength, and wavenumber by
the following equalities :

E = hv = = hc

where h is Plancks constant, which has a value of 6.626 1034
J .s.
Common types of Spectroscopy
X-ray
X-rays of sufficient energy are used to excite the inner shell
electrons in the atoms of a sample. The electrons move to outer
orbitals then down into the vacated inner shells and the energy in
this de-excitation process is emitted as radiation.
The absorption or emission energies are characteristic of the
specific atom and small energy variations may occur that are
characteristic of particular chemical bonding. The X-ray
frequencies can be measured and X-ray absorption and emission
spectroscopy is used to determine elemental composition and
chemical bonding.
Common types of Spectroscopy
Flame
Usually the analyte is in solution form (or converted into one) that
is then converted to a free gaseous form in a multistage process
(atomization). This method is often used for metallic element
analytes present at very low concentration ranges.
Common types of Spectroscopy
Visible/Ultraviolet (UV)
This uses the fact that many atoms are able to emit or absorb
visible light. The atoms must be in a gaseous phase in order to
obtain a spectrum just as those obtained in flame spectroscopy. It
is common for visible absorption spectroscopy to be combined
with UV absorption spectroscopy in UV/Vis spectroscopy.
UV spectroscopy can also be used to analyze fluorescence from a
sample in a form of absorption spectroscopy.
Common types of Spectroscopy
Infrared (IR) and Near Infrared (NIR)
IR spectroscopy is used to show what types of bonds are present
in a sample by measuring different types of inter-atomic bond
vibrations at different frequencies. It relies on the fact that
molecules absorb specific frequencies which is dependent on
their chemical structure. This is determined by factors such as the
masses of the atoms.
NIR shows a greater penetration depth into a sample than mid-
infrared radiation. This indicates a low sensitivity but also that it
allows large samples to be measured in each scan by NIR
spectroscopy with little (if any) sample preparation. It has
numerous practical applications that include: medical diagnosis
pharmaceuticals, biotechnology, various analyses (genomics,
proteomic) and chemical imaging of intact organisms, textiles,
forensic lab application and various military applications.
Common types of Spectroscopy
Nuclear Magnetic Resonance
This is a prominent method for analyzing organic compounds
because it exploits the magnetic properties of certain atomic
nuclei to determine the properties (both chemical and physical) of
these atoms or the molecules containing them.
It can provide extensive information about the structure,
dynamics, and chemical environment of atoms. Additionally, even
different functional groups are distinguishable, and identical
functional groups in differing molecular environments still give
distinguishable signals.
Spectroscopic Analysis
Atomic Spectroscopy
Molecular Spectroscopy
Atomic Spectroscopy
the determination of elemental composition by its electromagnetic
or mass spectrum.
Electrons exist in energy levels within an atom. These levels have well
defined energies and electrons moving between them must absorb
or emit energy equal to the difference between them.
The science of atomic spectroscopy has yielded three techniques for
analytical use: Atomic Absorption, Atomic Emission, Atomic
Fluorescence. The process of excitation and decay to the ground
state is involved in all three fields of atomic spectroscopy. Either the
energy absorbed in the excitation process, or the energy emitted in
the decay process is measured and used for analytical purposes.
Atomic Spectroscopy
Atomic Absorption
measures the amount of light at the resonant wavelength which
is absorbed as it passes through a cloud of atoms. As the number
of atoms in the light path increases, the amount of light absorbed
increases in a predictable way. By measuring the amount of light
absorbed, a quantitative determination of the amount of analyte
element present can be made.
atomic absorption is the most widely applied of the three
techniques and usually offers several advantages over the other
two, particular benefits may be gained with either emission or
fluorescence in special analytical situations.
Atomic Spectroscopy
Atomic Emission
a sample is subjected to a high energy, thermal environment in
order to produce excited state atoms, capable of emitting light.
The energy source can be an electrical arc, a flame, or more
recently, a plasma.
The emission spectrum of an element exposed to such an energy
source consists of a collection of the allowable emission
wavelengths, commonly called emission lines, because of the
discrete nature of the emitted wavelengths.
Atomic Spectroscopy
Atomic Fluorescence
This technique incorporates aspects of both atomic absorption
and atomic emission.
The intensity of this "fluorescence" increases with increasing
atom concentration, providing the basis for quantitative
determination.
Molecular Spectroscopy
the study of absorption of light by molecules.
a given molecule will specifically absorb only those wavelengths
which have energies that correspond to the energy difference of the
transition that is occurring.
In the gas phase at low pressures, molecules exhibit absorption in
narrow lines which are very characteristic of the molecule as well as
the temperature and pressure of its environment. In the microwave
and long-wavelength infrared regions of the spectrum, these lines are
due to quantized rotational motion of the molecule. At shorter
wavelengths similar lines are due to quantized vibration and
electronic motion as well as rotational motion.
Instruments for measuring
Absorption

INSTRUMENTS USED FOR MOLECULAR UV/VIS ABSORPTION

INSTRUMENTS USED FOR INFRARED ABSORPTION
INSTRUMENTAL COMPONENTS OF UV/VIS
ABSORPTION
Sources of UV radiation
It is important that the power of the radiation source does not
change abruptly over it's wavelength range.
The electrical excitation of deuterium or hydrogen at low pressure
produces a continuous UV spectrum. The mechanism for this
involves formation of an excited molecular species, which breaks
up to give two atomic species and an ultraviolet photon.
INSTRUMENTAL COMPONENTS OF UV/VIS
ABSORPTION
Sources of visible radiation
The tungsten filament lamp is commonly employed as a source of
visible light. This type of lamp is used in the wavelength range of
350 - 2500 nm. The energy emitted by a tungsten filament lamp is
proportional to the fourth power of the operating voltage. This
means that for the energy output to be stable, the voltage to the
lamp must be very stable indeed. Electronic voltage regulators or
constant-voltage transformers are used to ensure this stability.
INSTRUMENTAL COMPONENTS OF UV/VIS
ABSORPTION
Wavelength selector (monochromator)
All monochromators contain the following component parts;
An entrance slit
A collimating lens
A dispersing device (usually a prism or a grating)
A focusing lens
An exit slit
Polychromatic radiation (radiation of more than one wavelength) enters
the monochromator through the entrance slit. The beam is collimated,
and then strikes the dispersing element at an angle. The beam is split into
its component wavelengths by the grating or prism. By moving the
dispersing element or the exit slit, radiation of only a particular
wavelength leaves the monochromator through the exit slit.
INSTRUMENTAL COMPONENTS OF UV/VIS
ABSORPTION
Cuvettes
The containers for the sample and reference solution must be
transparent to the radiation which will pass through them. Quartz
or fused silica cuvettes are required for spectroscopy in the UV
region. These cells are also transparent in the visible region.
Silicate glasses can be used for the manufacture of cuvettes for
use between 350 and 2000 nm.
INSTRUMENTAL COMPONENTS OF UV/VIS
ABSORPTION
Detectors
photomultiplier tube is a commonly used detector in UV-Vis
spectroscopy. It consists of a photoemissive cathode (a cathode
which emits electrons when struck by photons of radiation),
several dynodes (which emit several electrons for each electron
striking them) and an anode.
linear photodiode array is an example of a multichannel photon
detector. These detectors are capable of measuring all elements
of a beam of dispersed radiation simultaneously. A linear
photodiode array comprises many small silicon photodiodes
formed on a single silicon chip.
INSTRUMENTS USED FOR MOLECULAR UV/VIS
ABSORPTION
Filter Photometer
The simplest instrument for molecular UV/Vis absorption is a
filter photometer which uses an absorption or interference filter
to isolate a band of radiation. The filter is placed between the
source and the sample to prevent the sample from decomposing
when exposed to higher energy radiation. A filter photometer has
a single optical path between the source and detector, and is
called a single-beam instrument. The instrument is calibrated to
0% while using a shutter to block the source radiation from the
detector.
INSTRUMENTS USED FOR MOLECULAR UV/VIS
ABSORPTION
Single-beam Spetrophotometer
An instrument that uses a monochromator for wavelength selection
The simplest spectrophotometer is a single-beam instrument equipped
with a fixed-wavelength monochromator Single-beam
spectrophotometers are calibrated and used in the same manner as a
photometer.
The accuracy of a single-beam spectrophotometer is limited by the
stability of its source and detector over time.
INSTRUMENTS USED FOR MOLECULAR UV/VIS
ABSORPTION
Double-Beam Spectrophotometer
The limitations of fixed-wavelength, single-beam
spectrophotometers are minimized by using a double-beam
spectrophotometer.
A scanning monochromator allows for the automated recording
of spectra. Double-beam instruments are more versatile than
single-beam instruments, being useful for both quantitative and
qualitative analyses, but also are more expensive.
INSTRUMENTS USED FOR MOLECULAR UV/VIS
ABSORPTION
Diode Array Spectrometer
An instrument with a single detector can monitor only one
wavelength at a time. If we replace a single photomultiplier with
many photodiodes, we can use the resulting array of detectors to
record an entire spectrum simultaneously in as little as 0.1 s. In a
diode array spectrometer the source radiation passes through the
sample and is dispersed by a grating.
One advantage of a diode array spectrometer is the speed of data
acquisition, which allows to collect several spectra for a single
sample.
One disadvantage of a photodiode array is that the effective
bandwidth per diode is roughly an order of magnitude larger than
that for a high quality monochromator.
INSTRUMENTS USED FOR MOLECULAR UV/VIS
ABSORPTION
Sample Cells
The sample compartment provides a light-tight environment that
limits the addition of stray radiation. Samples are normally in the
liquid or solution state, and are placed in cells constructed with
UV/Vis transparent materials, such as quartz, glass, and plastic.
INSTRUMENTS USED FOR INFRARED
ABSORPTION
Filter Photometer
The simplest instrument for IR absorption spectroscopy is a filter
photometer similar for UV/Vis absorption. These instruments
have the advantage of portability, and typically are used as
dedicated analyzers for gases such as HCN and CO.
INSTRUMENTS USED FOR INFRARED
ABSORPTION
Double-beam spectrophotometer
Infrared instruments using a monochromator for wavelength
selection use double-beam optics. Double-beam optics are
preferred over single-beam optics because the sources and
detectors for infrared radiation are less stable than those for
UV/Vis radiation. In addition, it is easier to correct for the
absorption of infrared radiation by atmospheric CO2 and H2O
vapor when using double-beam optics
INSTRUMENTS USED FOR INFRARED
ABSORPTION
Fourier transform spectrometer
In a Fourier transform infrared spectrometer, or FTIR, the
monochromator is replaced with an interferometer. Because an
FT-IR includes only a single optical path, it is necessary to collect a
separate spectrum to compensate for the absorbance of
atmospheric CO2 and H2O vapor. This is done by collecting a
background spectrum without the sample and storing the result
in the instruments computer memory. The background spectrum
is removed from the samples spectrum by ratioing the two
signals. In comparison to other instrument designs, an FTIR
provides for rapid data acquisition, allowing an enhancement in
signal-to-noise ratio through signal-averaging.
INSTRUMENTS USED FOR INFRARED
ABSORPTION
Sample Cells
Infrared spectroscopy is routinely used to analyze gas, liquid, and
solid samples. Sample cells are made from materials, such as NaCl
and KBr, that are transparent to infrared radiation. Gases are
analyzed using a cell with a pathlength of approximately 10 cm.
Longer pathlengths are obtained by using mirrors to pass the
beam of radiation through the sample several times.
Volatile liquids must be placed in a sealed cell to prevent their
evaporation.