Raman Spectroscopy

During a voyage to Europe in 1921, Raman noticed

the blue color of sky was more brighter when
passing through the Mediterranean sea. He was
motivated to discover the reason for the blue color.

Why sky is blue ?

The “rods” are our low-light night-vision black/white photo-receptors. They contain
“rhodopsin” photochemical which is sensitive to very low light levels and so the rods are all
fully bleached-out (non-functioning) in full daylight. It takes about 25mins for all of the
rhodopsin to recombine after lights out… why it takes us that long for our eyes to “get used
to the dark” (the cones restore their photochemicals quicker, in about 5minutes)
 K. S. Krishnan
Raman carried out experiments regarding the scattering of light by water and transparent
blocks of ice which explained the phenomenon.

In his first report to Nature, titled "A New Type of Secondary Radiation," Raman indicated
that approximately 60 different liquids had been studied, and all showed the same result
— some scattered light had a different color than the incident light. "It is thus," Raman
said, "a phenomenon whose universal nature has to be recognized.“
The Raman Effect is a very weak effect; only one in a million of the scattered light
particles, or photons, actually exhibits the change in wavelength. This explains, in part,
why the effect was not discovered earlier.

Raman employed monochromatic light from a mercury arc lamp which penetrated
transparent material and was allowed to fall on a spectrograph to record its spectrum. He
detected lines in the spectrum, which were later called Raman lines.
He presented his theory at a meeting of scientists in Bangalore on 16 March 1928, and
won the Nobel Prize in Physics in 1930.

In Munich, some physicists were initially unable to reproduce Raman's results, leading to
skepticism. However, Peter Pringsheim was the first German to reproduce Raman's results
successfully and sent his spectra to Arnold Sommerfeld. Pringsheim was the first to coin
the term "Raman effect" and "Raman lines."
The first commercial Raman instrument using dispersive
gratings and laser as an excitation source, was introduced
as early as 1966, according to the literature, following the
advent of the laser in the 1960s.
In addition to photons of EM radiation being absorbed and emitted by atoms and molecules,
photons also get scattered by them, although the probability is 1 in 107 Molecule/atoms.

1. Elastic scattering: Photons interacts with molecules and leaves the molecule in the same
state. The energy of photons remain same ( Rayleigh scattering)

2. In-elastic scattering: Photons interacts with molecules and leaves the molecule in the
altered state. The energy of photons may increase or decrease (Raman scattering)

Classical Theory of Raman Spectrum

where EE denotes the strength of the electric field and α is the polarizability of the
atom or molecule with units

This induced dipole moment is related to the polarizability of the molecule or atom and
the strength of the electric field by the following equation
A polarizable molecule in an electric field will have an induced dipole that oscillates:
m = a [E0 sin (2pnt)]
When a molecule is in an electric field E, the electron cloud and nuclei become polarized
resulting in an induced dipole moment P. The size of the dipole moment induced by a field of
magnitude E is given by the polarizability α of the molecule:

(α has units of volume). Light consists of oscillating electric and magnetic fields. For light of
frequency ν0, the magnitude of the electric field may be written:

Thus, the induced dipole moment oscillates in phase with the applied field:

The polarizability will depend upon the geometry of the molecule; as the molecule vibrates,
the polarizability will change. If we write the polarizability as a Taylor series expansion in a
nuclear coordinate r about its equilibrium position (r0)

Alternatively, we can expand α in terms of qi, the vibrational displacement coordinate of the
ith normal mode. The classical normal mode vibration is
 The intensity of the incident light I0 is proportional to E0 2 , while the scattered power is
proportional to |P|2 . The first term in above equation represents the Rayleigh scattering.
The last two terms oscillate at the sum and difference frequencies, and represent the anti-
Stokes and Stokes scattering, respectively.
The intensity of the Raman scattering is
seen to be proportional to the derivative of
the polarizability (squared):

The intensity of the Raman scattered radiation

IR is given by:

where Io is the incident laser intensity, N is the number of scattering molecules in a given state, n is the
frequency of the exciting laser, a is the polarizability of the molecules, and Q is the vibrational amplitude
The molecular vibration (or rotation) must cause some change in the polarizabiity of
different values along different the molecule.

𝑃𝑥 𝛼𝑥𝑥 𝛼𝑥𝑦 𝛼𝑥𝑧 𝐸𝑥

𝑃𝑦 = 𝛼𝑦𝑥 𝛼𝑦𝑦 𝛼𝑦𝑧 𝐸𝑦
𝑃𝑧 𝛼𝑧𝑥 𝛼𝑧𝑦 𝛼𝑧𝑧 𝐸𝑧

αxy, for example, is the dipole moment along the x-direction due to a unit electric field
polarized along the y-direction. The polarizability tensor, α is defined by these nine
coefficients.

However, because αxy = αyx, αzx = αxz, αyz = αzy there are only six independent
coefficients. The six coefficients may be used to define an ellipsoid:

The polarizability of a non-isotropic molecule is divided

into three orthogonal components, and the values of
these components determine the dimensions of the
polarizability ellipsoid.
 During each of the three modes of vibrations of H 2O, symmetric stretching, mode (v1),
bending mode (v2) and antisymmetric stretching mode (v3), the polarizability changes and
hence all the three are Raman active

In the case of C02 molecule, it is seen that during symmetric stretching (v1), polarizabiity
changes because of the change in ellipsoid size and this mode is Raman active. For the
bending (v2) and antisymmetric stretching modes (v3), there is no overall change in
polarizability ellipsoid. Hence, the bending and antisymmetric stretching modes of C02 are
Raman inactive
Polarizability
Neutral nonpolar species have spherically symmetric arrangements
of electrons in their electron clouds. When in the presence of an
electric field, their electron clouds can be distorted. The ease of
this distortion is defined as the polarizability of the atom or
molecule. The created distortion of the electron cloud causes the
originally nonpolar molecule or atom to acquire a dipole moment.
This induced dipole moment is related to the polarizability of the
molecule or atom and the strength of the electric field by the
following equation

NOTE: Except for the case of spherical symmetry, the

magnitude of the induced dipole moment depends
on the direction of the electric field.

For example, in case of a diatomic molecule, the

induce dipole moment will be higher in magnitude
when the electric field direction is along the inter-
nuclear axis than that of the perpendicular to the
inter-nuclear axis.
Factors that Influence Polarizability

The relationship between polarizability and the factors of electron density, atomic radii, and
molecular orientation is as follows:
1.The greater the number of electrons, the less control the nuclear charge has on charge
distribution, and thus the increased polarizability of the atom.

2.The greater the distance of electrons from nuclear charge, the less control the nuclear
charge has on the charge distribution, and thus the increased polarizability of the atom.

3.Molecular orientation with respect to an electric field can affect polarizibility (labeled
Orientation-dependent), except for molecules that are: tetrahedral, octahedral or
icosahedral (labeled Orientation-independent). This factor is more important for
unsaturated molecules that contain areas of electron-dense regions, such as 2,4-hexadiene.
Greatest polarizability in these molecules is achieved when the electric field is applied
parallel to the molecule rather than perpendicular to the molecule.
4. The extent of polarizability of a molecule in an electric
field depends upon the direction of the electric field a)
parallel b) perpendicular relative to the molecule, if the
same polarizabilty, then isotropic molecule or else
anisotropic
Polarisability Change during Rotation

In case of a diatomic and linear molecule, the induce dipole moment will be higher in
magnitude when the electric field direction is along the inter-nuclear axis than that of the
perpendicular to the inter-nuclear axis.
Quantum Theory of Raman Spectrum
Pure Rotational Raman Spectra

The rotational energy levels for a linear molecule are given

where J = 0,1,2, etc, B is the rotational constant for the molecule and εJ is the
rotational energy expressed in wavenumbers.
The selection rule for rotational Raman spectra is

The transition ΔJ = 0 means that the scattered radiation has the same frequency as the
incident radiation and it appears as Rayleigh scattering only.

It is worth recollecting that the selection rule for microwave spectra is ΔJ = ±1.
Rotational Raman spectra, the selection rule is ΔJ = 0, ±2. We know that the rotational
changer in the Raman spectrum depend on the polarizability of the molecule. The
polarizability in turn is associated with two dipole transitions - one for the incoming,
and one for the outgoing photon. Hence, there will be two quantum rotational jumps
as given by ΔJ = 0, ±2
When Δ J = +2, we can write for a transition J→ J + 2
where J = 0,1,2,3, ....... and it is the rotational quantum number in the lower state

for rotational spectra

, is the Raman shift for the rotational Raman spectra or the rotational Raman shift

We can understand that +ΔVrot stands for anti-Stokes lines while -ΔVrot stands for Stokes
lines
There is a general rule for labelling any series of lines in the spectrum which states that
when Δ J = +2, + 1, 0, -1, - 2, the corresponding group of frequencies are called S, R, Q, P
and 0 branches of the spectrum, respectively.

Thus in the above case, when ΔJ = +2, we shall observe S-branch of the spectrum. Notice
that when J = 0 is substituted , the separation of the first line from the exciting line is
found to be 6B. Once the value of rotational constant B is known, we can evaluate the
moment of inertia of the molecule since,
Rotational Raman spectra for molecules like H2, 02, HCN, CO, CO2 etc. have lines would be 48.
yielded useful information about their molecular structure. This information is especially
useful for homonuclear diatomic molecules which give no infrared or microwave spectra.

It should be mentioned that if the molecule has a centre of symmetry (as in the case of H2,
N2, 02, CO2 the nuclear spin exerts its effect in the Raman and IR spectra. The intensity
pattern can be explained using nuclear spin effect. We shall not go into the theoretical
background of this aspect but state a few examples. The significant conclusion is that the
intensity pattern of rotational Raman spectra has a vital role to play while providing data
regarding molecular structure.

1) The rotational Raman spectra of C02(g) showed a series of absorption peaks separated by
3.16 cm-1 in the S branch. What is the value of the rotational constant (B) for C02 molecule ?
In the case of C02, +ΔVrot , = 8B

2) For hydrogen molecule, the spacing between the S-branch lines in the Raman spectrum is
243.2 cm-1 . Calculate the bond length of hydrogen. Mass of hydrogen atom = 1.673 x 10-27
kg. For hydrogen molecule, the spacing between the lines (ΔV) = 4B
For vibrational Raman spectroscopy, the gross selection rule is that the polarizability of the
molecule should change as it vibrates. The specific selection rule for vibrational Raman
spectroscopy is ∆v = ±1, where the ∆v = 1 corresponds to Stokes lines and the ∆v = −1
corresponds to Anti-Stokes lines.
Vibrational spectroscopy spectrum rules

Gross selection rule in IR spectroscopy: vibration must lead to an oscillating dipole

Gross selection rule in Raman spectroscopy: vibration must lead to a change in
polarizability

• Vibrations that have a symmetric polarizability change do not appear in Raman.

• Bending vibrations are not strong Raman modes.
• Symmetric stretches tend to be strong Raman modes.

Combined use of IR and Raman to work out

structure
In general, vibrations can be in both IR and
Raman but those
which are strong in IR tend to be weak in
Raman, and vice
versa
 Advantages of Raman over IR
1. Very suitable for biological samples in
native state (because water can be used
as solvent).
2. Water can be used as solvent.
3. Although Raman spectra result from
molecular vibrations at IR frequencies,
spectrum is obtained using visible light
or NIR radiation. =>Glass and quartz
lenses, cells, and optical fibers can be
used. Standard detectors can be used.
4. Few intense overtones and combination
bands => few spectral overlaps.
5. Totally symmetric vibrations are
observable.
6. Raman intensities a to concentration and
laser power.
Advantages of IR over Raman
1. Simpler and cheaper
instrumentation.
2. Less instrument dependent than
Raman spectra because IR
spectra are based on
measurement of intensity ratio.
3. Lower detection limit than
(normal) Raman.
4. Background fluorescence can
overwhelm Raman.
5. More suitable for vibrations of
bonds with very low polarizability
(e.g. C–F).
Choose Raman Spectroscopy when:
•Investigating carbon bonds in aliphatic and aromatic rings are of primary interest
•Bonds that are difficult to see in FTIR (i.e., 0-0, S-H, C=S, N=N, C=C etc.)
•Examination of particles in solution is important, e.g. polymorphism
•Lower frequency modes are important (e.g. Inorganic-Oxides)
•Reactions in aqueous media are investigated
•Reactions in which observation through a reaction window is easier and safer (e.g. high
pressure catalytic reactions, polymerizations)
•Investigating lower frequency lattice modes is of interest
•Investigation of reaction initiation, endpoint, and product stability of biphasic and colloidal
reactions

Choose FTIR Spectroscopy when:

•Studying liquid-phase reactions
•Reactions in which reactants, reagents, solvents and reaction species fluoresce
•Bonds with strong dipole changes are important (e.g. C=O, O-H, N=O)
•Reactions in which reagents and reactants are at low concentration
•Reactions in which solvent bands are strong in Raman and can swamp key species signal
•Reactions in which intermediates that form are IR active
 Raman Spectroscopy Instrumentation

Optical filters – these optical

components are placed in
the Raman beam path, and are
used to selectively block the laser
line (Rayleigh scatter) whilst
allowing the Raman scattered https://core.ac.uk/download/pdf/80838463.pdf
light through to
the spectrometer and detector.
 Mass Spectrometry

Mass spectrometry is a powerful analytical technique used to quantify known materials, to identify
unknown compounds within a sample, and to elucidate the structure and chemical properties of
different molecules. The complete process involves the conversion of the sample into gaseous ions, with
or without fragmentation, which are then characterized by their mass to charge ratios (m/z) and relative
abundances.

Basic Principle

A mass spectrometer generates multiple ions from the sample under investigation, it then separates
them according to their specific mass-to-charge ratio (m/z), and then records the relative abundance of
each ion type.

• The first step in the mass spectrometric analysis of compounds is the production of gas phase ions of
the compound, basically by electron ionization. This molecular ion undergoes fragmentation. Each
primary product ion derived from the molecular ion, in turn, undergoes fragmentation, and so on.
• The ions are separated in the mass spectrometer according to their mass-to-charge ratio, and are
detected in proportion to their abundance.
• A mass spectrum of the molecule is thus produced. It displays the result in the form of a plot of ion
abundance versus mass-to-charge ratio. Ions provide information concerning the nature and the
structure of their precursor molecule.
• In the spectrum of a pure compound, the molecular ion, if present, appears at the highest value of
m/z (followed by ions containing heavier isotopes) and gives the molecular mass of the compound.
The instrument consists of three major components:
1.Ion Source: For producing gaseous ions from the substance being studied.
2.Analyzer: For resolving the ions into their characteristics mass components according to their mass-to-
charge ratio.
3.Detector System: For detecting the ions and recording the relative abundance of each of the resolved
ionic species.

Ions are very reactive and short-lived, their

formation and manipulation must be conducted
in a vacuum. The pressure under which ions
may be handled is roughly 10-5 to 10-8 torr (less
than a billionth of an atmosphere). Atmospheric
pressure is around 760 torr (mm of mercury).

Cations formed by the electron bombardment

(red dots) are pushed away by a charged
repeller plate (anions are attracted to it), and
accelerated toward other electrodes, having
slits through which the ions pass as a beam.
Some of these ions fragment into smaller
cations and neutral fragments. Ionization is
effected by a high energy beam of electrons,
and ion separation is achieved by accelerating
and focusing the ions in a beam, which is then
bent by an external magnetic field.
When a high energy electron collides with a molecule it often ionizes it by knocking away one of the
molecular electrons (either bonding or non-bonding). This leaves behind a molecular ion (colored red in
the following diagram). Residual energy from the collision may cause the molecular ion to fragment into
neutral pieces (colored green) and smaller fragment ions (colored pink and orange). The molecular ion is
a radical cation, but the fragment ions may either be radical cations (pink) or carbocations (orange),
depending on the nature of the neutral fragment.

A perpendicular magnetic field deflects the ion beam in an arc whose radius is proportional to the mass
of each ion. Lighter ions are deflected more than heavier ions. By varying the strength of the magnetic
field, ions of different mass can be focused progressively on a detector fixed at the end of a curved tube
(also under a high vacuum).

The ions are then detected electronically and the resulting information is stored and analyzed in a
computer.

A mass spectrum will usually be presented as a vertical bar graph, in which each bar represents an ion
having a specific mass-to-charge ratio (m/z) and the length of the bar indicates the relative abundance of
the ion. The most intense ion is assigned an abundance of 100, and it is referred to as the base peak.
Most of the ions formed in a mass spectrometer have a single charge, so the m/z value is equivalent to
mass itself. The highest-mass ion in a spectrum is normally considered to be the molecular ion, and
lower-mass ions are fragments from the molecular ion, assuming the sample is a single pure compound.
The mass spectra of three simple gaseous compounds, carbon dioxide, propane and cyclopropane. The
molecules of these compounds are similar in size, CO and C H both have a nominal mass of 44 amu, and
2 3 8

C H has a mass of 42 amu. The molecular ion is the strongest ion in the spectra of CO and C H , and it is
3 6 2 3 6

moderately strong in propane. The unit mass resolution is readily apparent in these spectra (note the
separation of ions having m/z=39, 40, 41 and 42 in the cyclopropane spectrum). Even though these
compounds are very similar in size, it is a simple matter to identify them from their individual mass
spectra. Even with simple compounds like these, it should be noted that it is rarely possible to explain
the origin of all the fragment ions in a spectrum. Also, the structure of most fragment ions is seldom
known with certainty.
Fragmentation Patterns

The nature of the fragments often provides a clue to the molecular structure, but if the molecular ion has
a lifetime of less than a few microseconds it will not survive long enough to be observed. Without a
molecular ion peak as a reference, the difficulty of interpreting a mass spectrum increases markedly.
Fortunately, most organic compounds give mass spectra that include a molecular ion, and those that do
not often respond successfully to the use of milder ionization conditions. Among simple organic
compounds, the most stable molecular ions are those from aromatic rings, other conjugated pi-electron
systems and cycloalkanes. Alcohols, ethers and highly branched alkanes generally show the greatest
tendency toward fragmentation.

The presence of a functional group, particularly one having a

heteroatom Y with non-bonding valence electrons (Y = N, O, S,
X etc.), can dramatically alter the fragmentation pattern of a
compound. This influence is thought to occur because of a
"localization" of the radical cation component of the molecular
ion on the heteroatom. After all, it is easier to remove (ionize) a
non-bonding electron than one that is part of a covalent bond.
By localizing the reactive moiety, certain fragmentation
processes will be favored. These are summarized in the
following diagram, where the green shaded box at the top
displays examples of such "localized" molecular ions. The first
two fragmentation paths lead to even-electron ions, and the
elimination (path #3) gives an odd-electron ion. Note the use of
different curved arrows to show single electron shifts
compared with electron pair shifts.
The complexity of fragmentation patterns has led to mass spectra being used as "fingerprints" for
identifying compounds. Environmental pollutants, pesticide residues on food, and controlled substance
identification are but a few examples of this application.

The following mass spectrum of cocaine demonstrates how a forensic laboratory might determine the
nature of an unknown street drug.

High Resolution Mass Spectrometry

By designing mass spectrometers that can determine
m/z values accurately to four decimal places, it is
possible to distinguish different formulas having the
same nominal mass.
 Nuclear magnetic resonance spectroscopy
Nuclear magnetic resonance spectroscopy, commonly referred to as nmr, has become the prominent
technique for determining the structure of organic compounds.

Typical Applications of NMR:

1.) Structural (chemical) elucidation
‚ Natural product chemistry
‚ Synthetic organic chemistry
- analytical tool of choice of synthetic chemists
- used in conjunction with MS and IR
2.) Study of dynamic processes
‚ reaction kinetics
‚ study of equilibrium (chemical or structural)
3.) Structural (three-dimensional) studies
‚ Proteins, Protein-ligand complexes
‚ DNA, RNA, Protein/DNA complexes
‚ Polysaccharides
4.) Metabolomics
5.) Drug Design
‚ Structure Activity Relationships by
NMR Structural Biology
6.) Medicine -MRI
A moving perpendicular An electric current in a closed loop will create
external magnetic field a perpendicular magnetic field
will induce an electric
current in a closed loop

Magnetic moment
The spinning nuclei possess angular
momentum, P, and charge, and so an
associated magnetic moment, .

m = p

The gyromagnetic ratio (also sometimes

Similar to magnetic
known as the magnetogyric ratio in other
field created by
disciplines) of a particle or system is
electric current flowing
the ratio of its magnetic moment to
in a coil
its angular momentum
The spin states of the nucleus
are quantified: I, (I - 1), (I - 2), NMR “active” Nuclear Spin (I)
… , -I = ½:
1H, 13C, 15N, 19F, 31P

I= 1/2 (e.g. 1H) biological and chemical

relevance
Odd atomic mass
I = +½ & -½

NMR “inactive” Nuclear Spin

(I) = 0:
12C, 16O

Even atomic mass &

number
In the ground state all nuclear spins are
disordered, and there is no energy difference
between them. They are degenerate.
Magnet
α – spin state
Since they have a magnetic moment, when we
apply a strong external magnetic field (Bo),
they orient either against or with it: There is
always a small excess of nuclei (population
excess) aligned with the field than pointing
against it
Magnet
β – spin state
 The applied magnetic field causes an energy difference between the aligned (a) and unaligned
(b) nuclei

NMR signal results from the transition of spins from the a to b state

Strength of the signal depends on the population difference between the a and b spin states

The population (N) difference can be determined from the Boltzmann distribution and the
energy separation between the a and b spin states:

Na / Nb = e DE / kT
Any free system with a constant gyromagnetic ratio, such as a rigid system of charges,
a nucleus, or an electron, when placed in an external magnetic field B (measured in teslas)
that is not aligned with its magnetic moment, will precess at a frequency f (measured
in hertz), that is proportional to the external field:
Since: Na / Nb =
DE = hn 1.000060
and
n =  Bo / 2p
then: Very Small !
~60 excess
The DE for 1H at 400 MHz (Bo = 9.39 T) is 6 x 10-5 Kcal / mol spins per
million in lower
Na / Nb = e DE / kT Na /Nb = e(hBo/2pkT) state
NMR signal (s) depends on:

1) Number of Nuclei (N) (limited to field homogeneity and filling factor)

2) Gyromagnetic ratio (in practice  3)

3) Inversely to temperature (T)

4) External magnetic field (Bo2/3, in practice, homogeneity)

5) B12 exciting field strength (RF pulse)

Na / Nb = e DE / kT

DE =  h Bo / 2p

Increase energy gap ->

Increase population
difference -> Increase
NMR signal

~$800,000 ~$2,00,000 ~$4,500,000

 Magnet Beffective = Bapplied - Blocal

α – spin state of a nuclei if it is only proton with

no electron cloud,

Hence, Blocal = 0
α–
spin
Beffective is having high value,
state
so DE would be large enough to bring 180 o
spin

Magnet Beffective = Bapplied - Blocal

α – spin state of a nuclei if it is proton

e- e- surrounded by high electron cloud,
e-
e- Hence, Blocal = +ve value
intermedi
ate – spin e- e-
state e- Beffective is having low value,
e-
e- e- so DE would be small enough to bring 180 o spin
 Magnet

α – spin state
Magnet
a

β – spin state
Magnet

e- e-
intermedi e- b
ate – spin e-
state e- e-
e- e-
e- e-
ClCH2CH3

Unknown chemical
Mass spectroscopy: C3H7Cl
IR spectroscopy = peak at CH vibration
Degree of unsaturation = 0 ( through analysis)
Thus, the probable structure is

1) 1-cloropropane (ClCH2CH2CH3)
2) 2-cloropropane (CH2CHClCH2)

If NMR is performed
• Electronegative atoms pulls the
electron cloud towards itself
• As the distance increases, the
effect decreases
 Solving NMR spectra
Nuclear Magnetic Resonance (NMR) interpretation plays a pivotal role in molecular identifications.
As interpreting NMR spectra, the structure of an unknown compound, as well as known
structures, can be assigned by several factors such as chemical shift, spin multiplicity, coupling
constants, and integration.

Strategy for Solving Structure

Here is the general strategy for solving structure with NMR:
1.Molecular formula is determined by chemical analysis such as elementary analysis
2.Double-bond equivalent (also known as Degree of Unsaturation) is calculated by a simple
equation to estimate the number of the multiple bonds and rings. It assumes that oxygen (O)
and sulfur (S) are ignored and halogen (Cl, Br) and nitrogen is replaced by CH. The resulting
empirical formula is C H
a b

3.Structure fragmentation is determined by chemical shift, spin multiplicity, integral

(peak area), and coupling constants (1J1J, 2J2J)
4.Molecular skeleton is built up using 2-dimensional NMR spectroscopy.
5.Relative configuration is predicted by coupling constant ( J). 3
 Chemical Shift
Chemical shift is associated with the Larmor frequency of a nuclear spin to its chemical environment.
Tetramethylsilane (TMS, (CH3)4Si(CH3)4Si) is generally used as an internal standard to determine
chemical shift of compounds: δTMS=0 ppm. In other words, frequencies for chemicals are measured for
a 1H or 13C nucleus of a sample from the 1H or 13C resonance of TMS. It is important to understand trend
of chemical shift in terms of NMR interpretation. The proton NMR chemical shift is affect by nearness to
electronegative atoms (O, N, halogen.) and unsaturated groups (C=C,C=O, aromatic). Electronegative
groups move to the down field (left; increase in ppm). Unsaturated groups shift to downfield (left) when
affecting nucleus is in the plane of the unsaturation, but reverse shift takes place in the regions above and
below this plane. 1 H chemical shift play a role in identifying many functional groups. Figure 11. indicates
important example to figure out the functional groups.

Chemical equivalence
Protons with Chemical equivalence has
the same chemical shift due to
symmetry within molecule
(CH3COCH2CH2COCH3) or fast
rotation around single bond (-CH3;
methyl groups).
Spin-Spin Splitting

Spin-Spin splitting means that an absorbing peak is split by more than one “neighbor” proton.
Splitting signals are separated to J Hz, where is called the coupling constant. The spitting is a
very essential part to obtain exact information about the number of the neighboring protons.
The maximum of distance for splitting is three bonds. Chemical equivalent protons do not result
in spin-spin splitting. When a proton splits, the proton’s chemical shift is determined in the
center of the splitting lines.
Spin Multiplicity (Splitting pattern)
Spin Multiplicity plays a role in determining the number of neighboring protons. Here is a
multiplicity rules: In case of AmBnAmBn system, the multiplicity rule is that Nuclei of BB element
produce a splitting the AA signal into nB+1nB+1 lines. The general formula which applies to all nuclei
is 2nI+12nI+1, where II is the spin quantum number of the coupled element. The relative intensities of
the each lines are given by the coefficients of the Pascal’s triangle (Figure 22).
the spectrum for 1,1,2-trichloroethane
ethyl acetate:
Problems
Draw the 1H NMR spectrum for 2-Hydroxypropane in CDCl3. Assume sufficient resolution to
provide a first-order spectrum and ignore vicinal proton-proton coupling(3JHH)

Solution
1) the structure of 2-hydoroxyporpane is drawn

Figure out which protons are chemically equivalent, i.e., two methyl (-CH3) groups are
chemical equivalent.

Figure1): chemical shift of methyl groups (H a) : 1-2 ppm (?Ha=1.1 ppm); chemical shift of -CH-
groups (Hb) moves to downfield due to effect on aldehyde groups:2-3ppm ( ?Hb=2.4 ppm);
chemical shift of aldehyde groups (Hc):9-10 ppm (?Hc=9.6 ppm)
4) Splitting pattern is determined by (N+1) rule: Ha is split into two peaks by Hb(#of
proton=1). Hb has the septet pattern by Ha (#of proton=6). Hc has one peak.(Note
that Hc has doublet pattern by Hb due to vicinal proton-proton coupling.)
1.Sample holder: Glass tube with 8.5 cm long, 0.3 cm in diameter.
2.Permanent magnet: It provides homogeneous magnetic field at 60-100 MHZ
3.Magnetic coils: These coils induce magnetic field when current flows through them
4.Sweep generator: To produce the equal amount of magnetic field pass through the sample
5.Radio frequency transmitter: A radio transmitter coil transmitter that produces a short
powerful pulse of radio waves
6.Radio frequency receiver: A radio receiver coil that detects radio frequencies emitted as
nuclei relax to a lower energy level
7.Read out systems: A computer that analyses and record the data.

https ://microbenotes.com/nuclear-magnetic-resonance-nmr-spectroscopy/
Limitations of 1D NMR spectroscopy

• It is not possible to correlate two different chemical shifts by 1D NMR

• It is difficult to analyze a 1D NMR spectrum in case of large molecules due
to overlap of peaks

CH3-CH2----------------CH2CH3
 What signals do you expect to see in the 1H NMR spectrum of 1,1-dichloroethane
CH3CHCl2?

How many signals do you expect to see in the 1H NMR spectra of 2-bromopropane (CH3)2CHBr and 1-
bromopropane CH3CH2CH2Br?

A compound A analyses as containing 69.52 % C and 10.21 % H. Among other absorptions,

the IR spectrum of A contains a sharp band at 2250 cm–1. The 1H NMR spectrum (recorded
in CDCl3), two triplets (δ 2.34, 1.08 ppm) and a sextet (δ 1.70 ppm) are observed.
Compound A is:

In the IR spectrum of a compound X, there is a strong absorption at 1718 cm–1 in addition

to bands at 2978 and 2940 cm–1 and bands below 1500 cm–1. The 1H NMR spectrum
contains two signals: a quartet and a triplet with relative integrals of 2 : 3. Of the following
compounds, which is X most likely to be?
Laser Induced Breakdown Spectroscopy (LIBS) is a rapid chemical analysis technology that uses a short
laser pulse to create a micro-plasma on the sample surface. This analytical technique offers many
compelling advantages compared to other elemental analysis techniques. These include:
• A sample preparation-free measurement experience
• Extremely fast measurement time, usually a few seconds, for a single spot analysis
• Broad elemental coverage, including lighter elements, such as H, Be, Li, C, N, O, Na, and Mg
• Versatile sampling protocols that include fast raster of the sample surface and depth profiling
• Thin-sample analysis without the worry of the substrate interference

A typical detection limit of LIBS for heavy metallic elements is in the low-PPM range. LIBS is applicable
to a wide range of sample matrices that include metals, semiconductors, glasses, biological tissues,
insulators, plastics, soils, plants, soils, thin-paint coating, and electronic materials.

Initiation of a high-temperature (>15,000K) plasma

above the sample surface and a rapid expansion of the
plasma into the ambient medium.
Emission of continuum light during early stage (< Emission of discrete atomic lines at later times (>
200 ~ 300 nsec) of plasma cooling process. 1 µsec).

So how does LIBS work?

The main physical process that forms the essence of LIBS technology is the formation of high-
temperature plasma, induced by a short laser pulse.

When the short-pulse laser beam is focused onto the sample surface, a small volume of the
sample mass is ablated (i.e. removed via both thermal and non-thermal mechanisms) — in a
process known as Laser Ablation.
This ablated mass further interacts with a trailing portion of the laser pulse to form a highly
energetic plasma that contains free electronics, excited atoms and ions. Many fundamental
research projects have shown that the plasma temperature can exceed 30,000K in its early
life time phase

When the laser pulse terminates, the plasma starts to cool. During the plasma cooling
process, the electrons of the atoms and ions at the excited electronic states fall down into
natural ground states, causing the plasma to emit light with discrete spectral peaks.

The emitted light from the plasma is collected and coupled with an ICCD/spectrograph
detector module for LIBS spectral analysis. Each element in the periodic table is associated
with unique LIBS spectral peaks. By identifying different peaks for the analyzed samples, its
chemical composition can be rapidly determined. Often, information on LIBS peak intensities
can be used to quantify the concentration of trace and major elements in the sample.

With the advancement of powerful chemometric software for LIBS data analysis, and with
steady progress in understanding laser ablation fundamentals, today’s analytical researchers
are applying LIBS effectively for both quantitative and material discriminatory analysis for a
wide range of sample matrices.
 Display of LIBS spectra and their subsequent
Emitted light collection by a set of optical lens
analysis by the instrument software for both
and optical fiber
qualitative and quantitative elemental analysis