UV/VIS & IR

Spectroscopy

Dr. M. Coley
 Spectroscopy
• the measurement of EM radiation
that is absorbed, scattered or
emitted by atoms, molecules or
other chemical species.
• Exception? …… mass spectroscopy
 Types of Spectroscopy
 UV/VIS:
• Absorption of radiation
causes electronic transitions.

 Infrared:
• Absorption of radiation
causes vibrational
transitions.
• Radiation absorbed causes
bonds to vibrate, rotate,
stretch, wag, bend, twist,
rock, etc.
• Different functional groups will
show characteristic behaviour
depending on the structure of
the molecule.
Infra-red Spectroscopy
 Interacts with
molecules to cause
stretching, bending,
rotation or vibration of
bonds.

 Characteristic
vibrations may be used
to identify various
functional groups.

 This type of
spectroscopy is very
useful for probing the
structure of organic
molecules.
 Vibrational Transitions

C – H symmetrical C – H Asymmetric C – H In-plane scissoring

stretching 2925 cm-1 stretching: 2850 cm-1 1480 - 1440 cm-1

Out-of-plane twisting In-plane rocking Out-of-plane wagging

 IR Data Correlation Table
Functional Group Wavenumber, cm-1
Alcohol O-H 3650-3200
Acid O-H 3300-2500*
Phenol O-H 3600-3000*
Acid C=O 1725-1700
Ketone C=O 1725-1705
Aldehyde C=O 1740-1720
Ester C=O 1750-1730
Amide C=O 1690-1680
Primary amine N-H (stretch) Two bands 3500 and 3300
N-H (deform) 1650-1580
Secondary amine N-H (stretch) 3450-3300
N-H (deform) 1650-1550
Aromatic C=C (stretch) 1600
Alkene C=C (stretch) 1680-1620**
Alkyne CC 2260-2100
Nitrile CN 2400-2200
Aromatic C-H 3150-3000
Aliphatic C-H 3000-2850
* O-H stretches are broad due to hydrogen bonding
** weak bands
 Gas Phase IR Spectrum
of Formaldehyde
symmetric

asymmetric

Out-of-plane
wagging
 UV/VIS Spectroscopy
 is the application of visible or ultraviolet
light in the identification or quantification
of analyte species

 applicable to a wide range of species:

– Metals – Fe, Mn, Cr
– Non-metals – PO43-, NO3-, SiO4 ,organics
– Any species that can absorb a measurable amount
of UV or visible radiation resulting in an atomic or
molecular electronic transition
 Electronic Transitions
Eo E1

Photon of
energy:
E = hv
E = hc/λ
 Properties of EM Radiation
 Energy of a photon may be represented as:
E = hv
Since: c = vλ
and v = c/λ

 Hence: E = hc/λ

 Also: E = E1 – Eo = hc/λ

E is the energy responsible for transition of an electron from a ground

state (Eo) to an excited state (E1). The intensity of the energy, E,
absorbed by species in a homogeneous solution is proportional to the
 Beer-Lambert’s Law
 “The amount of light absorbed by a
homogeneous solution of absorbing species that
is exposed to monochromatic light is directly
proportional to the number of absorbing species
present in the light path.” ie: A = Ecl

A = Ecl
compare:
y = mx

Higher sample concentration, greater absorbance produced.

 Calculating Absorbance &
Transmittance
A = ─Log T = Log 1/T
If transmittance, T = 0.25, If absorbance is 0.015,
(i.e. 25%) calculate the calculate transmittance.
absorbance. A = 0.015
T = 0.25 0.015 = -Log T
A = log 1/T Log T = -0.015

A = log 1/0.25 T = 0.966

T = 96.6 %
A = 0.602
 Determining Concentrations by
UV-Vis spectroscopy
Prepare a series of standards  For coloured species, simply dilute
of known concentrations. standards and samples.

Measure absorbances of CrO42- Cr2O72-

standards at λmax.
Measure absorbance of
unknown at same wavelength.
 For colourless species, add suitable
colouring agent, eg. KSCN.
 Intensity of coloured complex is then
directly proportional to analyte concn.
 Standards & samples should contain
same amount of colouring agent &
other additives (acids, etc.)
 Beer’s law requires pathlength in
units of cm & concentration: mol L-1.
Preparation of Standards

λmax

“Use the data to plot a graph.”

Using Beer’s law to determine
Sample Concentration
Spectrophotmetric
Determination of Iron
Fe(II) conc.
Solution %T Absorbance
(mg/L)

blank 0.000 100 0.000

1 0.102 94 0.056

2 0.511 77 0.119

3 1.02 60 0.194 Sample =

1.63 mg/ml
4 2.55 23 0.509

5 5.11 9 1.046

unknown ? 42 0.377
 Beer’s Law - Applications
 Example:
 A series of standard solutions containing Fe(III) concentrations as
shown in the table below, were made up in the presence of excess potassium
thiocyanate (KSCN) solution. A sample of unknown Fe(III) concentration
was developed similarly and when measured in a 1 cm cuvette, gave an
absorbance of 0.652.
 i. From a graph, determine the concentration of the original sample
solution assuming it was diluted by a factor of 10 prior to analysis.

 ii. Estimate the value of the molar absorptivity, E.

0.652 unknown
 Beer’s Law - Applications
Absorbance of dilute
1.5 Sample = 0.652
y = 0.7792x
Absorbance

1
R2 = 0.9995 [dilute Sample]
= 0.836 mol L-1
0.5

0
0 0.5 1 1.5
Concentration Mol/L

From graph: [Fe(II)] = 0.836 mol L-1 Concn of Original Sample

= 10/1 x 0.836 mol L-1
Recall: A = Ecl = 8.36 mol L-1
 0.652 = E x 0.836 mol L-1 x 1cm
 E = 0.652/(0.836 mol L-1 x 1cm)
 E = 0.779 mol-1 L cm-1
 Problem
 Following a 1:10 dilution, a solution was measured
at 820 nm to determine its PO43- concentration.
If the absorbance, A is 0.500, the path-length, l is 20mm
and the molar absorptivity, E is 100 M-1cm-1, what is the
concentration of the original solution?

 A = Ecl
 c = A/(E x l)

= 0.500/100 M-1cm-1 x 2 cm
= 0.0025M
Considering dilution factor:
Original concn: = 10 x 0.025M
= 0.025M
Instrumentation:
Single-beam spectrophotometer
Spectrophotometer–
Double Beam
 Questions
 Question 1 Recall: A = Ecl
Beer's Law states that:
a) absorbance is proportional to both the path length and the
concentration of the absorbing species;
b) absorbance is proportional to the log of the concentration
of the absorbing species
c) the analyte solution should be heterogeneous.
Question 2
IR spectroscopy is based on electronic transition(s) while UV-Vis
Spectroscopy requires vibrational transitions? TRUE? FALSE?
• Question 3
The detector is the dispersing component used in a single-beam
spectrophotometer. TRUE? FALSE?
• Question 4
UV/Vis spectroscopy requires monochromatic radiation while
IR spectroscopy requires polychromatic light. TRUE? FALSE?
• Question 5
Both UV/VIS & IR spectroscopy can be used for analyte identification.
EXERCISE YOUR
MIND