Analytical Chemistry

Lecture 6
Spectroscopic methods of analysis –
Part 2
Absorption of ultraviolet, visible and infrared
radiation
Instructor: Nguyen Thao Trang
Molecular absorption
• Three types of energy changes occur when molecules are
excited by ultraviolet, visible, and infrared radiation.
• Energy change due to electronic transition:
– Upon being excited, an electron of a molecule residing in a low-energy
molecular or atomic orbital is promoted to a higher-energy orbital.

• The change in energy levels is

Excited called a transition.
state
• When a transition occurs:
Energy hν of the absorbed photon
= Energy difference between the
two orbital energies.
Ground
state
• The transition of an electron
between different energy levels
is called an electronic transition.
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Molecular absorption
• Energy change due to vibrational transition:
– Upon being excited, a molecule can undergo vibrational transition.

– Vibrational transition occurs when a molecule has multiple quantized

energy levels (vibrational states) associated with the bonds that hold
a molecule together.

– Differences in energy among the vibrational states are significantly

smaller than among energy levels of the electronic states. 3
Molecular absorption
• Energy change due to rotational transition:
– Upon being excited, a molecule can undergo rotational transition.

– Rotational transition occurs when a molecule has multiple quantized

rotational energy levels associated with the rotational movements of
the molecule around its center.

– Rotational energy states are superimposed on each of the vibrational

states shown in the energy diagram.

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Molecular absorption
• Energy-level diagram for a molecule: being absorbed infrared,
visible and ultraviolet radiation is shown:

Vibrational states are

Energies E1 and E2 indicated by the
of the first two of lighter horizontal
the several lines labeled 1, 2, 3,4.
electronically
excited states . The lowest
vibrational levels are
labeled 0.

Energy IR radiation is
not sufficient to
Energy E0 of the cause an electronic
ground state. transition.
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Absorption of ultraviolet (UV) and visible
(Vis) radiation

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UV and Vis absorption
• Absorption of UV and visible light by a molecule causes
electronic transitions.

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Review: Covalent bonds
• Sigma bond (σ): A bond where the line of electron density is
concentrated symmetrically along the line connecting the two
atoms.
• “End to end” overlap .

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Review: Covalent bonds
• Pi bond (π): A bond where the overlapping regions exist above
and below the internuclear axis (with a nodal plane along the
internuclear axis).
• “Side by side” overlap.

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Review: Covalent bonds
• In a multiple bond, the first bond is a sigma (σ) bond and the
second and third bonds are pi (π) bonds.
π π
π

C C C C

σ σ
..
• Non-bonded pairs (n): O: n
σ π

H3C CH3 10
Review: Covalent bonds
• Bonding and antibonding:
– Sigma bonding:

Antibonding

Higher energy

Bonding
Lower energy

Increased electron density

in the internuclear region

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Review: Covalent bonds
• Bonding and antibonding:
– Pi bonding:

Antibonding

Higher energy

Bonding
Lower energy

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Electronic transitions
• Electronic energy levels for polyatomic molecule:

σ * (antibonding)

π * (antibonding)

Energy n (non-bonding)
n σ* n π*

π (bonding)
π π*
σ (bonding)
σ σ*
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Electronic transitions
σ σ∗ alkanes 150 nm

π π∗ carbonyls 180 nm

π π∗ unsaturated compounds 180 nm √ - if conjugated!

n σ∗ O, N, S, halogens 190 nm

n π∗ carbonyls 300 nm √

• Routine organic UV spectra are typically collected from 200-700 nm.

• Special equipment to study vacuum (< 100 nm) or far UV is

required.

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Absorption of some functional groups

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Instrumentation
• The construction of a traditional UV-VIS spectrometer
requires:
– Light source

– λ - selection

– Sample container

– Detector

– Signal processing

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Instrumentation
• Light source:

D2 lamp (UV: 160 – 375 nm) W lamp (Vis: 350 – 2500 nm)

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Instrumentation
• Light source:

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Instrumentation
• λ - selection:
– UV and visible regions are usually equipped with one or more devices
to restrict the radiation being measured to a narrow band à enhance
selectivity and the sensitivity of an instrument.

– Isolate the desired wavelength band by:

• Monochromator

• Filter

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Instrumentation
• λ - selection:
– Monochromator: isolate the desired wavelength band by
• Employ a diffraction grating to disperse the radiation into its
component wavelengths.
• Rotating the grating, different wavelengths can be made to pass
through an exit slit.
• Can be used for spectral scanning applications or at a fixed
wavelength.

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Instrumentation
• λ - selection:
– Filter:
• Interference filters used for absorption measurements.
• These filters transmit radiation over a bandwidth of 5 - 20 nm.
Radiation outside the transmitted bandpass is removed by
destructive interference.
• Only used for fixed wavelength.

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Instrumentation
• Sample container: cuvettes
– b = 1 cm typically

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Instrumentation
• Sample container: cuvettes
– b = 1 cm typically
– Glass (Vis) or fused silica (UV 350 nm)

Working range for different optical materials

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Instrumentation
• Detector :
– Used to detect the radiant power transmitted.
– Photon detector is based on the interaction of radiation with a
reactive surface to produce electrons or to promote electrons to
energy states in which that can conduct electricity.

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Instrumentation
• UV-Vis spectrometer:

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Absorption of infrared (IR) radiation

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IR absorption
• IR photons have low energy. The only transitions that have
comparable energy differences are molecular vibrations and
rotations.

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IR absorption
• In order for IR absorbance to occur two conditions must be
met:

– A change in the dipole moment of the molecule as a result of a

molecular vibration (or rotation).

– If the frequency of the radiation matches the natural frequency of the

vibration (or rotation), the IR photon is absorbed and the amplitude of
the vibration increases.

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IR absorption
• There are three types of molecular transitions that occur in IR:
1. Rotational transitions
- When an asymmetric molecule rotates about its center of mass, the
dipole moment seems to fluctuate.
- ΔE for these transitions correspond to 𝜈̅ < 100 cm-1
- Quite low energy, show up as sharp lines that subdivide vibrational
peaks in gas phase spectra.

2. Vibrational and rotational transitions

- Arise from changes in the molecular dipole moment due to the
combination of a bond vibration and molecular rotation.

3. Vibrational transitions
- The most important transitions observed in qualitative mid-IR
spectroscopy with 𝜈̅ = 13,000 – 675 cm-1 (0.78 – 15 µm).

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IR absorption
• Vibrational modes:
1. Stretching: the rhythmic movement along a bond axis with a
subsequent increase and decrease in bond length

2. Bending: a change in bond angle or movement of a group of atoms

with respect to the rest of the molecule.
In-plane bending Out of plane bending

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IR absorption
• Vibrational modes:
– Non-linear molecule : 3n – 6 vibrational modes and 3 possible
rotational modes.

– Linear molecule : 3n – 5 vibrational modes and 2 possible rotational

modes.
(n is the number of atoms in the molecule, and 3n cartesian
coordinates are called as degree of freedom).

– Example:
Linear CO2 : 3n – 5 = 3 ×3 – 5 = 4
Nonlinear H2O : 3n – 6 = 3 ×3 – 6 = 3
HCHO : 3n – 6 = 3 ×4 – 6 = 6

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IR absorption
• Vibrational modes of CO2:

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IR absorption
• Vibrational modes of H2O:

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IR absorption
• Vibrational modes of H2O:

~ 1596 cm-1
Bending

~ 3650 – 3750 cm-1

Symmetric and asymmetric stretch

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 Characteristics of IR absorption
Intensity: s = strong, m = medium, w = weak, v = variable

Group Freq. Range (cm-1) Intensity

Alkyl
C–H (stretching) 2853–2962 (m–s)

Alkenyl
C–H (stretching) 3010–3095 (m)
C=C (stretching) 1620–1680 (v)

Alkynyl
≡C–H (stretching) ~3300 (s)
C≡C (stretching) 2100–2260 (v)

Aromatic
Ar–H (stretching) 3000 - 3300 (v) sharp

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 Characteristics of IR absorption
Intensity: s = strong, m = medium, w = weak, v = variable

Group Freq. Range (cm-1) Intensity

Alcohols, Phenols & Carboxylic Acids

O–H (stretching)
- alcohols & phenols 3200–3550 (broad, s)
(hydrogen bonded)
- carboxylic acids 2500–3000 (broad, v)
(hydrogen bonded)

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Characteristics of IR absorption
Intensity: s = strong, m = medium, w = weak, v = variable

Group Freq. Range (cm-1) Intensity

Aldehydes, Ketones, Esters, Carboxylic Acids, Amides
C=O (stretching) 1630–1780 (s)
Aldehydes 1690–1740 (s)
Ketones 1680–1750 (s)
Esters 1735–1750 (s)
Carboxylic Acids 1710–1780 (s)
Amides 1630–1690 (s)

Amines
N–H 3300–3500 (m)

Nitriles
C≡N 2220–2260 (m) 37
General regions of a IR spectrum

Blue bands: stretching vibrations.

Green band: bending vibrations.
Fingerprint region: < 1400 cm-1 (C-C, C-O, C-N) arise from
complex deformations of the molecule.
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Interpretation of an IR spectrum
• Step 1: Look for the carbonyl C=O band at 1820-1660 cm-1.
– This band is usually the most intense absorption band , and has a
medium width. If you see the carbonyl band, look for other bands
associated with functional groups that contain the carbonyl in step 2.
– If no C=O present, go to step 3.

• Step 2: the carbonyl C=O band present, need to check if it is

part of an acid, aldehyde or ketone.
– Acid:
• Look for an O-H band at 3300-2500 cm-1 . This band overlaps the C-
H stretch.
• Also a C-O single bond band near 1100-1300 cm-1.
• Look for the carbonyl band near 1725-1700 cm-1.

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Interpretation of an IR spectrum
• Step 2: the carbonyl C=O band present, need to check if it is
part of an acid, aldehyde or ketone.
– Ester:
• Look for C-O absorption of medium intensity near 1300-1000 cm-1.
• There will be no O-H band.

– Aldehyde:
• Look for aldehyde type C-H absorption bands. These are two weak
absorptions to the right of the C-H stretch near 2850 cm-1 and
2750 cm-1 and are caused by the C-H bond that is part of the CHO
aldehyde functional group.
• Look for the carbonyl band around 1740-1720 cm-1.

– Ketone:
• The weak aldehyde C-H absorption bands will be absent.
• Look for the carbonyl CO band around 1725-1705 cm-1.
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Interpretation of an IR spectrum
• Step 3: If no carbonyl band, look for an alcohol O-H band.
– Alcohol:
• Look for the broad OH band near 3600-3300 cm-1.
• Look for a C-O absorption band near 1300-1000 cm-1.

• Step 4: If no carbonyl and O-H bands, look for C=C from an

aromatic or an alkene.
– Alkene:
• Look for weak absorption near 1650 cm-1 for a double bond.
• There will be a C-H stretch band near 3000 cm-1.

– Aromatic:
• Look for the benzene, C=C, double bonds which appear as medium
to strong absorptions in the region 1650-1450 cm-1.
• The C-H stretch band is much weaker than in alkenes.
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Interpretation of an IR spectrum
• Step 5: If none of the previous groups can be identified, you
may have an alkane.
– Alkane:
• The main absorption will be the C-H stretch near 3000 cm-1 . The
spectrum will be simple with another band near 1450 cm-1.

– C-H stretching region:

• Alkanes C-H stretch < 3000 cm-1
• Alkenes C-H stretch > 3000 cm-1
• Alkynes C-H stretch ~ 3300 cm-1

– C-H bending region:

• CH2 bending ~ 1460 cm-1
• CH3 bending (asym) appears near the same value
• CH3 bending (sym) ~ 1380 cm-1

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Interpretation of an IR spectrum
• Step 6: If the spectrum still cannot be assigned you may have
an alkyl halide.
– Alkyl bromide:
• Look for the C-H stretch
• Look for a relatively simple spectrum with an absorption to the
right of 667 cm-1.

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Interpretation of an IR spectrum
1-Hexene

Alkene C-H
C=C
stretch

Alkane C-H CH2 CH CH2 CH2 CH2 CH3

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Interpretation of an IR spectrum
Toluene

Alkane C-H

Alkene C-H
aromatic
C=C
CH3

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Interpretation of an IR spectrum
Exercise: Identify C-H and C=O functional groups

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Strength and limitations
• IR alone cannot determine a structure
• Some signals may be ambiguous
• The functional group is usually indicated
• The absence of a signal is definite proof that the functional
group is absent
• Correspondence with a known sample’s IR spectrum confirms
the identity of the compound

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Strength and limitations
• IR more difficult than UV-Vis because
– narrow bands (variation in ε)
– complex spectra
– weak incident beam
– low transducer sensitivity
– solvent absorption

• IR mostly used for rapid qualitative but not quantitative

analysis
– Long optical path-length required
– Beer’s law failure

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