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UNIT 4 Summary

• Regions of the electromagnetic spectrum that are used in spectroscopy:
• The sample solution absorbs electromagnetic radiation from an appropriate
source, and the amount absorbed is related to the [] of the analyte in the
solution.Example, the solution of copper ions is blue because it absorbs
complementary yellow colour.
• Spectroscopy is based on the absorption of photons by analyte.
• Spectrometry: measurement of radiation intensity as a function of wavelength
using electric transducer
• Photometry: measurement of (usually visible) radiation using a photoelectric
transducer (at a fixed wavelength)
• Spectrophotometry: photometry in which the measurement is performed as a
function of wavelength
• Spectrum: plots of intensity vs energy
• Electromagnetic spectrum: the wave described either in terms of its wavelength,
distance of one complete cycle, or in terms if frequency, the number of cycles
passing a fixed point per unit time. The shorter the wavelength or the greater the
frequency, the greater the energy.
• Most analytically used region is between 10-380 nm UV region
• IR is from 780nm to about 300000nm (or 600 to 4000 cm-1)
• How does matter absorb radiation?
• The colour of an object we see is due to the wavelength transmitted or reflected,
the other wavelengths are absorbed.
• When a polychromatic light (white light) is passed through an object, the object
will absorb certain of the wavelengths, leaving the unabsorbed wavelengths to be
transmitted. Therefore colour will be seen complementary to the absorbed one.
• There are three basic processes by which a molecule can absorb radiation: all
involve raising the molecule to a higher internal energy level, the increase in
energy being equal to the energy of the absorbed radiation, quantized(they exist at
discrete level).
• 1. rotational transition:. Fist the molecule rotates about various axis, the energy of
rotation being at definite energy level, thus the molecule may absorb radiation and
be raised to a higher rotational energy level.
• 2. vibrational transition: the atoms within the molecule vibrate relative to each
other, and the energy of this vibration occurs at definite quantize level.
• 3. electronic transition: the electrons of a molecule may be raised to a higher
energy
• The absorbing group in a molecule is called chromophores. An auxochrome does
notitself absorb radiation, but , if present in a molecule, it can inhance the
absorption by a chromophore or shift the wavelength of absorbtion when attached
to the chromophore.
• Beer’s Law:
• The absorption coefficient is directly proportional to the concentration of
absorbing species in solution for a fixed pathlength
• The amount of monochromatic radiation absorbed by a sample is described the
this law
• T=P/P0 transmittance
• Beers law describes the dependence of T on both the pathlength and the
concentration: T= P/P0= 10-ebc where e is a combined constant (molar absorptivity)
• Absorbance A=-log T =log 1/T=log P0/P= ebc
• Deviation from Beer law:
• It cannot always be assumed that beers law will apply that is a linear plot of
absorbance vs concentration
• Deviations occur as a result of chemical and instrumental factors
• True deviations will occur when the [] is so high that the index of refraction of the
solution is changed from that of the blank. A similar situation would apply for
mixtures of organic solvents with water. The solvent may also have an effect on
the absorptivity of the analyte.
• Apparent deviations may also occur when the substance can exist as a dimer as
well as a monomer. Equilibrium depends on the concentration. The best way to
minimize chemical deviations is by adequate buffering of pH, adding large excees
of complexing agent, ionic strength adjustment, etc. Preparation of the calibration
curve over the measurement range will correct for most deviations.
• If both species of a chemical equilibrium absorb, and if there is some overlap of
their absoption curves, the wavelength at which this occurs is called isosbestic
point, and the molar absorptivity is the same.
• The effect of pH could be eliminated by making measurements at the isosbestic
point, but the sensitivity is decreased.
• Instrumental deviations: page 505
• Other instrumental factors that contribute to the deviation of Beer Law include
stray radiation entering the monochromator and being detected
• Stray Light (any detected light that is not absorbed by the sample or is outside the
bandwidth of the selected wavelength) becomes especially limiting at high
absorbances and eventually causes deviation from linearity.
• Noise resulting from stray light also becomes a major contributor to the
spectrometric error or imprecision at high absorbances.
• So stray light : internal reflection in monochrometer , scaterring from sample
(optical curve), light leakage into instrument.
• Other chemical and instrumental sources on nonlinearity: hydrogen bonding,
interaction with the solvent, nonlinear detector, nonlinear electronics, etc.,
nonuniform cell thickness, air bubbles can affect path length and stray light.
• Appearance of spectra for UV:
• Effect of cell material and solvent on UV cut-off
• Bands are broader than the atomic spectra can be up to 100nm for aqueous
solution
• Instead of sharp individual bands, observe broad bands
• UV instrument:
• All spectrometers require: 1) a source of continuous radiation over the
wavelengths of interests, 2) a monochromator for selecting a narrow band of
wavelengths form the source spectrum, 3) a sample cell, 4) a detector, or
transducer, for converting radiant energy into electrical energy, 5) a device to read
out the response of the detector. Each of these will vary depending on the
wavelength region except readout device.

• Sources: should have a readily detectable output of radiation over the
wavelength
• for visible region: lamp is used, can be used for near-UV and near-infrared
regions
• for ultraviolet: a low pressure hydrogen or deuterium discharge tube is usually
used as a source , it must have a quartz window, because glass is not transparent
to ultraviolet radiation
• for infrared: so hot wires and light bulbs or glowing ceramics are used as sources,
nernst glower

• Monochromators (Selectors)
• Consists of lenses or mirrors to focus the radiation, entrance and exit slits to
restrict unwanted radiation
• Prisms – break the polychromatic lights into separate wavelengths , glass-visible,
quartz-UV, rocksalt-IR
• Diffraction grating- these consist of a large number of parallel lines ruled on a
highly polished surface such as aluminum. Dispersion by gratings is independent
of wavelength, but the intensity varies with wavelength
• The lines act as scattering centers for rays imprinting on the grating, the result is
equal dispersion of all wavelength of a given order (linear dispersion).
Particularly great for infrared spectrum
• Optical Filters- various types may be used to separate (isolate) certain
wavelength of light. Usually made of glass and dye o absorb all the colours except
for one to pass through

• Sample Cells:
• Must be transparent to the wavelength region being measured
• For visible and ultraviolet spectrometers are usually cuvets 1 cm thick , for
infrared plavtes such as NaCl are used

• Detectors:
• UV- a phototube (photocell) is used , as well as in visible, can be very specific for
different regions of the spectrum. Also, photomultiplier tube – more sensitive than
phototube. Third type id CCD charge-coupled device
• IR- poses property of heat, and heat detectors that transduce heat into an electrical
signal can be used. Thermocouples and bolometers are used. For rapid
measurements with FTIR instruments, and for high sensitivity measurements,
photon detectors are used.

• IR:
• Absorbing (vibrating) groups in the infrared region absorbs withing a certain
wavelength region, and exact wavelength will be influenced by the neighbouring
groups. Absorbtion peaks are so much sharper than UV, each molecule has a
fingerprint region (specific for each molecule).
• Most important use of IR is identification and structure analysis, it could also be
used for quantitative analysis of complex mixtures of similar compounds (with
intensities proportional to the concentration of absorbing species).
• Can have rotational/vibrational –(see notes above) transitions (far-infrared),
• For Vibrational modes:
• Quantitative analysis:
• Beer-Lambert Law generally applies:
• Determine peak baseline
• Measure height
• Convert to absorbance units
• A=-log T=-log (%T/100)
• Quantitation relies on pathlength
• -use liquid cell
• -use internal standard (resolve IR peaks)
• -plot Astd/AIS vs. Cstd.
• -add same amount of IS to sample
• -record Asample/AIS
• Two forms of IR :
• 1.) Dispersive
• -uses a grating monochromator
• -takes time to scan wavenumber range
• -need to calibrate wavenumber position using a standard

• 2.)Fourier Transform Infrared Spectroscopy (FTIR)
• -uses a interferometer
• -advantages of FTIR – greater throughput, increased signal –to-noise ration,
simultaneous measurement of all wavelengths.