LECTURE NOTE ON SPECTROSCOPY

CHAPTER TEN  Speed of light can be expressed as � where λ” is the wavelength;

10. Introductions to Spectroscopy �” is the frequency, and c the speed of light in a medium.
10.1 Introduction  wavenumber, ṽ” another unit used to describe the wave properties of
 Spectroscopy is a branch of science which studies the interaction of electromagnetic radiation which is the reciprocal of wavelength ṽ ⁄
 Particle Nature of light. When matter absorbs electromagnetic radiation it
electromagnetic radiation with matter where the interaction of radiation
with chemical species is measured to obtain characteristics quality and
undergoes a change in energy. The interaction between matter and
quantity of the species.
 Spectroscopic techniques can be divided in to two
electromagnetic radiation is easiest to understand if we assume that

 Atomic spectroscopy (atomic absorption and atomic emission

radiation consists of a beam of energetic particles called photons. When a
photon is absorbed by a sample it is “destroyed, and its energy acquired
spectroscopy)
 Molecular spectroscopy (Uv-Vis, IR, NMR, MS)
by the sample. The energy of a photon, in joules, is related to its
frequency, wavelength, and wavenumber.
10.1.1 Electromagnetic Radiation ṽ , where h is Planck's constant = 6.63 x 10–34 J s.
�
Electromagnetic radiation is a form of energy that is transmitted through
. � ∝ ṽ and ν but E ∝ ⁄�
space at enormous velocities. Electromagnetic radiation, or light, is described
by the properties of both waves and particles nature.
 Wave nature of light: In dealing with phenomena such as reflection,
Example 10-1. The energy difference between the 3p and the 3s orbitals in
a sodium atom is 2.107 eV. Calculate the � (in nm) that would be
refraction, interference, and diffraction, but electromagnetic radiation
absorbed in exciting the 3s electron to the 3p state (l eV = 1.60 X 10–19 J).
is conveniently modeled as waves.
An electromagnetic wave is characterized by several fundamental properties, Solution �
including its frequency, velocity, amplitude, phase angle, polarization, and
10.2 Electromagnetic radiation and its interactions with matter
direction of propagation.
 Frequency �”: the number of oscillations of the field that occurs per The interactions of radiation and matter are the subject of spectroscopic

second. studies. The most interesting types of interactions in spectroscopy are

 Wavelength, λ”: the linear distance between successive maxima or

absorption and emission of radiation by molecular or atomic species of
interest which involve transitions between different energy levels of the
minima of a wave. Mostly measured in nm = 10-9m
chemical species.

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Absorption and emission. The frequency and wavelength of electromagnetic radiation vary over many

 Absorption: When radiation passes through a transparent layer of a

orders of magnitude. For convenience, we divide electromagnetic radiation
into different regions. The electromagnetic spectrum-based on the type of
solid, liquid, or gas, certain frequencies may be selectively removed by atomic or molecular transition that gives rise to the absorption or emission of
the process of absorption. Here, electromagnetic energy is transferred to photons (Figure 10.2). The boundaries between the regions of the
the atoms or molecules constituting the sample; as a result, these electromagnetic spectrum are not rigid, and overlap between spectral regions
particles are promoted from a lower energy state to higher-energy is possible.
states, or excited states (Figure 10.1). Note that at room temperature,
most substances are in their lowest energy or ground state.
 Emission: when an atom or molecule in an excited state returns to a
lower energy state, the excess energy often is released as a photon, a
process we call emission (Figure 10.1).

Figure 10.1 Simplified energy diagram showing the absorption and

emission of a photon by an atom or a molecule.
.W ν , Figure 10.2 The electromagnetic spectrum showing the boundaries between
, Δ , different regions.
.
’ ’ ’ ’ ’ ’ ’ ’ ’ ’ ’
. , ν, the sample leads to a change in one or more of these characteristic properties.
Δ
The electromagnetic Spectrum .
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That means
 X-ray photons excite inner-shell electrons;
 ultra-violet and visible-light photons excite outer-shell (valence)
electrons;
 infrared photons are less energetic, and induce bond vibrations; and
 Microwaves are less energetic still, and induce molecular rotation.

10.3 Absorption Laws (Quantitative Analysis) Figure 10.3 (a) Schematic diagram showing the attenuation of radiation
passing through a sample; P0 is the radiant power from the source and PT is
As light traverses a medium containing an absorbing analyte, decreases in
the radiant power transmitted by the sample. (b) Schematic diagram showing
intensity occur as the analyte becomes excited.
 For an analyte solution of a given concentration, the longer the length
how we redefine P0 as the radiant power transmitted by the blank. Redefining
P0 in this way corrects the transmittance in (a) for the loss of radiation due
of the medium through which the light passes (path length of light),
’ ,’ ,’ ’ ’ ’ ’ ’ ’ ’
the more absorbers are in the path, and the greater the attenuation.
’ ’ ’
 Also, for a given path length of the light, the higher the concentration
matrix.

of absorbers, the stronger the attenuation.  An alternative method for expressing the attenuation of
 This attenuation of radiation is described quantitatively by two separate, electromagnetic radiation is absorbance, A, is defined as
but related terms: transmittance and absorbance. As shown in Figure
10.3 a, transmittance ’ ’ ’ ’ ’ ’ ’ ’ ing
the sample, PT, to that incident on the sample, P0. Absorbance is the more common unit for expressing the attenuation of radiation.
’ ’’ ’ ’ ’ ’ ’ .’

 Multiplying the transmittance by 100 gives the percent transmittance

Example 10.2 A sample has a percent transmittance of 50%. What is its
absorbance?
(%T), which varies between 100% (no absorption) and 0% (complete
Solution: A percent transmittance of 50.0% is the same as a transmittance of
absorption). All methods of detection, whether the human eye or a
modern photoelectric transducer, measure the transmittance of 0.500, and

electromagnetic radiation.

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. .’ ’ ’ :’ ’ W’ ’ ’ ’ ’ ’ ’ ’ ’ ’ ’
 The absorption law, also known as the Beer-Lambert law or just Beer's components. If there are no interactions between the components, the
law, tells us quantitatively how the amount of attenuation depends on individual absorbance, Ai, are additive. That is the total absorbance for a
the concentration of the absorbing molecules and the path length over multicomponent system at a single wavelength is the sum of the individual
which absorption occurs. absorbance.
 According to Beer's law, absorbance is directly proportional to the
concentration of the absorbing species c ’ ’ ’ ’ ’ b
of the absorbing medium, as expressed by following equation.
∝
 Here a is a proportionality constant called the absorptivity. Because
absorbance is a unit less quantity, the absorptivity must have units  ’ ’ ’
−1
that cancel the units of band c. If, for example, c has the units of gL ’ ’ ’ ’ ’ ’ ’ ’ ’ ’ ’
−1 −1
and b has the units of cm, absorptivity has the units of L g cm . If ’ .’ ’ ’
we express the concentration using molarity, then we replace a with 1. Instrumental deviation
the molar absorptivity ε”, which has units of cm–1 M–1 or L mol–1 cm–1.  Stray radiation (imperfection in λ selector)
 Polychromatic radiation (applied only in monochromatic radiation)
 ’ ’ ’ ’ ’ ’ ’ ’ ’ ’ 2. Chemical deviation
analyses.  Concentration deviation (applied only at low concentration)
–4
Example 10.3 A 5.00 × 10 M solution of an analyte is placed in a sample cell
with a path length of 1.00 cm. When measured at a wavelength of 490 nm, 10.4. Instruments for Optical Spectroscopy
’ ’ ’ ’ . 8.’W ’ ’ ’ ’ ’ ’ ’ The basic components of instruments for absorption spectroscopy, as well as

this wavelength? for emission and fluorescence spectroscopy; are remarkably alike in function

Solution and in general performance requirements whether the instruments are

designed for ultraviolet, visible, or infrared radiation. Because of the
Solving for ε and making appropriate substitutions gives
similarities, such instruments are frequently referred to as optical instruments
even though the eye is sensitive only to the visible region.

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 Generally, most spectroscopic instruments are made up of five

components while their configuration or arrangement may varies:





 , Figure 10.4 The arrangement of instruments components; (a) for absorption, (b) for
emission spectroscopy
 In absorption measurements (Figure 10.3a), source radiation of the 1. Sources of energy (radiation)
selected wavelength is sent through the sample, and the transmitted All forms of spectroscopy require a source of energy. In absorption
radiation is measured by the detector/signal processing/readout unit. spectroscopy this energy is supplied by photons. Emission spectroscopy uses

 In emission spectroscopy (Figure 10.3b), the sample itself is the emitter

thermal, radiant (photon), or chemical energy to promote the analyte to a less
stable, higher energy state.
and no external radiation source is needed. In these methods, the sample
is usually fed into plasma or a flame, which provides enough thermal A source of electromagnetic radiation must provide an output that is both
energy to cause the analyte to emit characteristic radiation. Here, a intense and stable in the desired region of the electromagnetic spectrum.
source of thermal energy, such as a flame or plasma, produces an analyte Sources of electromagnetic radiation are classified as either continuum
vapor that emits radiation isolated by the wavelength selector and or line sources.
converted to an electrical signal by the detector.  A continuum source emits radiation over a wide range of wavelengths,
with a relatively smooth variation in intensity as a function of wavelength.
 Line sources, on the other hand, emit radiation at a few selected, narrow
wavelength ranges.

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Table 10.1 Common Sources of Electromagnetic Radiation for  One limitation of an absorption or interference filter is that they do not
Spectroscopy allow for a continuous selection of wavelength. A further limitation is
that filters are available for only selected nominal ranges of wavelengths.
 Monochromator: provides a continuous variation of wavelength.
 As monochromator converts a polychromatic source of radiation at
the entrance slit to a monochromatic source of finite effective
bandwidth at the exit slit.
 Interferometer: a type of monochromator which allows sources
or radiation of all λ to reach the detector. (All λ are
measured simultaneously). It have two advantages
 Jacquinot advantage: results from the higher throughput of source
2. Wavelength selector
 We usually try to select a single wavelength where the analyte is the only
radiation. Since an interferometer does not use slits and has fewer
optical components from which radiation can be scattered and lost, the
absorbing species.
 Unfortunately, we cannot isolate a single wavelength of radiation from a
throughput of radiation reaching the detector is 80–200 times greater
than that achieved with a normal monochromator.
 Fellget advantage: reflects a savings in the time needed to obtain a
continuum source. Instead, we use a wavelength selector which passes a
narrow band of radiation.
 A wavelength selector passes a narrow band of radiation. Wavelength
spectrum. Since all frequencies are monitored simultaneously, an entire
spectrum can be recorded in approximately 1 s, as compared to 10–15
selector can be two types. Filter and monochromatic.
 Filter: The simplest method for isolating a narrow band of radiation. It
min with a scanning monochromator.

can be absorption or interference filter. 3. Sample containers

 Absorption filters work by selectively absorbing radiation from a  Sample containers are required for all spectroscopic studies except
narrow region of the electromagnetic spectrum. emission spectroscopic studies.
 Interference filters use constructive and destructive interference to  Sample containers, are usually called sample cells or cuvettes, and must
isolate a narrow range of wavelengths. have windows that are transparent in the spectral region of interest.

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 For example, quartz or fused silica is required for the UV region 5. Signal Processor and readout device
(wavelengths less than 350 nm) and may be used in the visible region  ’ ’ ’ ’ ’ ’ ’ ’ ’ ’ ’ ’ ’
’ ’ ’ ’ ’ ’ ’μ ’ ’ ’ ’ .’ displayed in a form that is more convenient for the analyst. Examples of
 Silicate glass is ordinarily used for the 375 to 2000-nm region because signal processors include analog or digital meters, recorders, and
of its low cost compared with quartz. Plastic cells are also used in the computers equipped with digital acquisition boards.
visible region.  ’ ’ ’ ’ ’ ’ ’ ’ ’ ’ ,’ ’
’ ’ ’ ,’ ’ ’ ’ ’ ,’ ’ ’
mathematically transform the signal.
 Finally, the processed signal displays on readout devices in a more
4. Detectors convenient form for the analyst.
In modern absorption and emission spectrometers, radiation
transducers are invariably used to convert light intensity into electrical 10.5 Atomic absorption and emission spectroscopy
signals that can be subsequently amplified, manipulated, and finally Atomic spectroscopy is based upon the absorption or emission of
converted into numbers proportional to the magnitude of light. Two electromagnetic radiation by atomic particles. Spectroscopic determination of
general types of radiation transducers are employed; one responds to atomic species can only be performed on a gaseous medium in which the
photons (photon transducers), and the other to heat (thermal individual atoms or elemental ions. This method is widely applied to a wide
transducers), several examples of which are listed in Table 10.4. range of metals and nonmetals.

Table 10.2 Common Detectors for Absorption Spectroscopy  The first step in all atomic spectroscopic procedures is atomization, a
process in which a sample is volatilized and decomposed to produce
gas-phase atoms and ions.
 Atomization is a critical step in all atomic spectroscopy. Several
methods are used to atomize samples for atomic spectroscopic studies.
E.g. inductively coupled plasmas, flames, and electrothermal atomizers;
Flames and electrothermal atomizers are widely used in atomic
absorption spectrometry, while the inductively coupled plasma is
employed in optical emission and in atomic mass spectrometry.

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 Atomic Absorption Spectroscopy (AAS):

 Is a type of elemental analysis based on excitation of electron of atom.

 There are two fundaments proses during application of

atomic absorption spectroscopy.

 Figure 10.5 Block diagram of the components of an AAS.
 ,
 The light source (hollow cathode lamp or electrodeless discharge
.
lamp) emits a spectrum specific to the element of which it is
.
analyzed, which is focused through the sample cell into the
monochromator. The anode and cathode are sealed in a glass cylinder
 Many samples like solid, animal tissue, plant leaves, minerals etc. are normally filled with either neon or argon at low pressure. At the end
not directly used as a sample, and rather they should be prepared as of the glass cylinder is a window transparent to the emitted radiation.
solution by extensive preliminary treatment.
 After the sample is prepared as a form of clear solution, it is introduced
to the instrument for atomization process. In the atomization proses
the sample is nebulized (conversion of samples to mist, i.e. small
droplets of solution) by a flow of gaseous oxidant (e.g. air, O2, N2)
Figure 10.6 hollow cathode lamp
mixed with gaseous fuel (e.g. natural gas, H2, ethylene (C2H2)).
Instrumentation When an electrical potential is applied between the anode and
 In the components of an atomic absorption or flame absorption cathode, some of the fill gas atoms are ionized. The positively charged
apparatus, the flame can be considered to be a dilute gaseous solution fill gas ions accelerate through the electrical field to collide with the
of the atomized sample held in place by the aspirator-burner. negatively charged cathode and dislodge individual metal atoms in a
Radiation from a suitable source is passed through the atomized ’ ’ .’ ’ ’ ’ ’then excited to
sample and into the slit of a photometer or spectrophotometer. an emission state through a kinetic energy transfer by impact with fill
gas ions.
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
suitable for the sample being analyzed. I.e. the expected sample
- concentration should be within the range established by the standard.
  The standards and the sample are then aspirated into the flame and the
- + -
: + → +

+
: + → + absorbance read from the instrument and related with concentration of


+ unknown sample using A= ac + b (where a and b are slop intercept of
: + → * +

the calibration curve respectively, c is concentration of unknown
: * → + �
 W
sample, and A is absorbance of Unknown sample read from the
*

instrument but a and b are determine using standard solution)
.

Application
AAS is an analytical technique used for the qualitative and quantitative
determination of the elements present in different samples like food, water
and wastewater sample, nanomaterial, biomaterials, forensics (blood
sample), and industrial wastes.
Figure 10.7 absorbance versus concentration calibration curve
1. Qualitative analysis: is based on color of flame test.
When alkali metals like calcium, strontium or barium salt is heated strongly In atomic emission spectroscopy (AES), in addition to liquid samples of
in the Bunsen flame, a characteristic flame color is observed: e.g. metal, nonmetallic solids can be analyzed.
Na yellow Comparison of AAS and AES
Li crimson Activities AAS AES
Ca brick red Process Absorption (light Emission (light emitted by
Sr crimson measured absorbed by exited atoms)
unexcited atoms)
Ba green
Use of flame Atomization Atomization and excitation
instrumentation Uses light source Do not use light source
2. Quantitative analysis:’ ’ ’ ’ .’ ’ ’ ’ ’ (independent of light source)
preparation of series of standard solution over a concentration range of ’ Applicable (A ∝ C) Not applicable (I = KC)
Data obtained A Vs C I Vs C

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10.6. Ultraviolet and Visible (UV-Vis) Spectroscopy  n → * and → * transitions: most common transitions observed in
 UV-Vis Spectrometry is based upon absorption of electromagnetic organic molecular UV-Vis, observed in compounds with lone pairs and
radiation in the visible and ultraviolet regions of the spectrum multiple bonds with λmax = 200-600 nm.
resulting in changes in the electronic structure of ions and molecules.  Of these transitions, the most important are the n → * and → *,
The wavelength of UV and visible light are substantially shorter than because they involve functional groups, or double/triple bonds that are
the wavelength of infrared radiation. The UV–Vis spectrum ranges characteristic of the analyte and wavelengths (Uv/vis) that are easily
from 200 to 700 nm. When a molecule or ion absorbs ultraviolet or accessible.
visible radiation it undergoes a change in its valence electron  Absorption of Uv-Vis radiation in organic molecules is restricted to certain
transition. functional groups (known as chromophores which are unsaturated organic
functional groups). Electrons involved in double and triple bonds of
organic molecules are not as strongly held and are therefore more easily
excited by Uv-Vis radiation. Uv-Vis radiation is not absorbed by saturated
organic molecule. This is because Uv-Vis radiation is less energetic to
excite electron from single bond.
Instrumentation
 Polychromatic light from the source is focused on the entrance slit of a
monochromator, which selectively transmits a narrow band of light. This
Figure 10.8 Relative energies of orbitals and possible transitions between them. light then passes through the sample area to the detector.
M + hv → M* (when the molecule exposed to radiation hv, valance  The absorbance of a sample is determined by measuring the intensity of
electron of the molecule is exited) light reaching the detector without the sample (the blank) and
The wavelength of light absorbed is that having the energy required to move comparing it with the intensity of light reaching the detector after
an electron from a lower energy level to a higher energy level. For example, passing through the sample. As discussed above, most


spectrophotometers contain two source lamps, a deuterium lamp and a
→ * transitions: high-energy, accessible in vacuum UV (λmax < 150
tungsten lamp, and use either photomultiplier tubes or, more recently,
nm). Not usually observed in molecular UV-Vis.
 n → * transitions: non-bonding electrons (lone pairs), wavelength
photodiodes as detectors.

(λmax) in the 150-250 nm region.

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10.7 Infrared spectroscopy (IR) basic principles

 The energy associated with IR radiation is sufficient to cause molecules to
rotate and to vibrate.
 There are two types of molecular vibrations or modes of vibration:
stretching and bending.
 In stretching vibration results in increasing or decreasing of bond length.
 In bending vibration causes a change in bond angle. E.g. Twisting,
Figure 10.9 Schematic of a conventional spectrophotometer Rocking, wagging, scissoring.
Application  All molecules do not absorb IR radiation. i.e., the molecule must have a
1. Qualitative analysis: change in dipole moment during vibration in order to absorb IR
 Have limited application to identify the functional group or particular radiation.
molecule as a result of absorption spectra.  The dipole moment μ is the product of the charge (Q) and the distance
 But in some extent it used to identify the unknown compound by between the charges (r). i.e. μ = Q x r
comparing the absorption spectra with known compound spectra.  E.g. on vibration of the HF, HCl, CO, the dipole moment changes
 E.g. a weak absorption band around 280-290nm indicates the presence because the distance between the charges changes. It is IR active.
O
of carbonyl ( C ) group, absorption band around 257nm
indicated existence of an aromatic ring.  Symmetrical diatomic molecules do not absorb IR radiation and called
2. Quantitative: as IR inactive. Example: H2, N 2 , O2, Cl2
 It is widely used for quantitative analysis. Because of its  Molecules like Carbon dioxide has two equal C=O bond dipoles but because
 Widely applicability to both organic and inorganic compounds the molecule is linear the bond dipoles cancel and the molecule has no net
 Typically sensitive (i.e. 10-4 to 10-5 M) and up to 10-7 with some dipole moment. Hence do not absorb IR radiation.
modification  Water molecule has two equal H-O bond dipoles. The water molecule has
 High sensitivity a permanent net dipole moment. It is IR active.
 Good accuracy
The quantitative figure is determined as ’ ’ ’ .

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10.8 Nuclear Magnetic Resonance Spectroscopy (NMRs) according to their mass-to-charge ratio, m/z, and measuring the
 NMR spectroscopy is based on the measurement of absorption of abundance of the ions formed.
electromagnetic radiation in the radio-frequency region of roughly 4 to  Mass spectrometry is a technique used for measuring the molecular
900 MHz, which has long wavelengths, and low energy and frequency. weight and determining the molecular formula of an organic
 When low-energy radio waves interact with a molecule, they can compound.
change the nuclear spins of some elements.  In a mass spectrometer, a molecule is vaporized and ionized by
 Nuclei of atoms rather than outer electrons are involved in the bombardment with a beam of high-energy electrons.
absorption process.  Mass spectrometer performs the following essential functions.
 It is a powerful analytical technique used to elucidating (characterize)  Firstly, it subjects molecules to bombardment by a stream of high-
organic molecules by identifying carbon-hydrogen frameworks within energy electrons, converting some of the molecules to ions, (ion source)
molecules.  Secondly, the mass Analyzer resolves (separates) the ions into their
 Two common types of NMR spectroscopy are used to characterize characteristics mass components according to their mass-to-charge ratio
organic structure: 1H NMR is used to determine the type and number in a magnetic or electric field.
’ ’ ’ ’ ’ ;’ C NMR is used to determine the type of  Thirdly, the abundance of the resolved ions that have a particular mass-
13

carbon atoms in the molecule. to-charge ratio are measure with a detector.
 When an external energy source (h) that matches the energy Finally, Process the signals from the detector that are transmitted to
difference (E) between these two states is applied, energy is absorbed, the computer and control the instrument using feedback

’ ’ ’ ’ ’ ’ ’ ’ ’ ’ .

10.9 Mass Spectrometry

 Mass spectrometry (MS) is a technique for creating gas phase ions
from the molecules or atoms in a sample, separating the ions

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