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UV/Vis-NIR absorption

spectroscopy
The Electromagnetic Spectrum
Wavewlength 103 102 101 100 10-1 10-2 10-3 10-4 10-5 10-6 10-7 10-8 10-9 10-10 1011 10-12
(metres)

Bacteria
Animal cells

Virus

Proteins
Longer H2O Shorter
Tennis ball
wavelength Building wavelength

Generic name
Radio waves Infrared ultraviolet Hard x-rays

Gamma rays
Microwave oven
Microwaves Soft x-rays

X-ray machine

radioactivity
Light bulb
Body heat
FM radio
AM radio

radar
Sources

Frequency (Hz) 106 107 108 109 1010 1011 1012 1013 1014 1015 1016 1017 1018 1019 1020

Energy of
a photon 10-9 10-8 10-7 10-6 10-5 10-4 10-3 10-2 10-1 100 101 102 103 5 105 106
(ev)
UV/Vis Spectroscopy

IR Spectroscopy
UV/Vis-NIR spectroscopy

What region of the EM spectrum are we interested in?

UVC 210-280 nm
UVB 280-320 nm
UVA 320-400 nm
Visible 400-700 nm

Far Red 700-1100 nm


Near-Infra red 1100-2500 nm (9000 cm-1- 4000 cm-1)
(increasingly important for materials science)
Mid Infrared 2500-6000 nm (4000 - 60 cm-1)
(unusual for electronic absorptions)
UV/Vis spectrum
1.4

•Plot of the absorbance (unitless dimension)


1.2 or molar absorptivity (M-1 cm-1) against
wavelength
1.0 •Expressed in nanometres (nm, 10-9 m)
•In older books and papers Angstroms (Å,
0.8
10-10 m) are used
•In the near infrared region (from 900-2500
Abs

nm) microns or micrometres (µm) are used


0.6

0.4

0.2

0.0
300 400 500 600 700 800
Wavelength /nm
UV region visible region far red region
Wavelength vs wavenumber 1.4

1.2
1.4
1.0

1.2
0.8

Abs
1.0
0.6

0.8 0.4
Abs

0.2
0.6

0.0
0.4 40000 35000 30000 25000 20000 15000
-1
wavenumber /cm
0.2

0.0
300 400 500 600 700 800
Wavelength /nm
Electronic Absorption
EM radiation has a magentic and an E = hν
electronic vector

If electrons move from one orbital to another


then there is a change in the charge
distribution in a molecule and hence a
change in the molecules electric dipole. λ
Selection rules
-Resonance condition
-Change in principle quantum number
-Change in orbital angular momentum
-No change in spin: (2S+1) must not change singlet singlet
-Laporte’s rule: transitions between orbitals
of the same parity are forbidden

singlet triplet
Electronic Absorption
• Total internal energy
Sum of the electronic,
vibrational and
rotational energy
• Etotal = Eelec + Evib + Erot
• For an electronic
transition:
∆Etotal= ∆Eelec+∆Evib+∆Erot
Molar Absorptivity
• Probability that a photon of light will be
absorbed upon passing through an
optically dilute solution
• Proportional to the square of the transition
moment (change in electron density)
• Forbidden transitions
Chromophores & Conjugation
Chromophore – brings colour
The absorption of light is quantised
Mixing of electronic, vibrational and
rotational energy levels leads to broad
spectra instead of lines

C=O C=C
Pigments & Dyes, I
• Retinal – cis/trans isomerisation is the first
step in sensing light
• Chlorophylls absorb light for
photosynthesis
• Types of transitions
n – π* (auxochromes have free electron pairs e.g C=ö: )
n – σ*
σ –σ*
π – π*
Vibrational progression
01
1.0 00 Frank-Condon state
Normaillised absorption and emission

0.8

02
0.6

9,10-di-phenyl-anthracene
0.4

0.2

0.0
28000 26000 24000 22000 20000
-1
wavenumber /cm
The Bougier-Beer-Lambert
Law
An Empirical law – i.e. based on
observation not theoretical
derivation
Transmittance
• The light passing through a sample
• Solvents and optical material are
transparent (100% transmittance) only in
certain regions of the EM spectrum
• Solvent cut-off (see table 7.6, Hesse):
– Water – 190
– CH3CN – 190
– Acetone -350
Transmittance
T = I1/I0
T = 10-A

ε
The Beer-Lambert-Bouguer Law
(empirical law)
• The intensity of light passing through a sample
decreases exponentially and the absorbance of light is
proportional to the concentration of the chromophore
• The absorbance of light is proportional to the pathlength
through which the light travels

A = εcl
A is absorbance, c is concentration (mol L-1), l is pathlength
(in cm) and ε is the molar absorptivity (L mol-1 cm-1)

N.B. Assumption that all particles behave independently


(no shadow effect)
Justification of BLB law
dI = - κ[J]I dl
where [J] is the molar concentration of the absorbing species, I is the incident intensity,
dI is the reduction in intensity of I, κ is the proportionality coefficient and dl is the
pathlength or thickness of the layer.

Rearranged the equation is


dI = - κ[J] dl
I
And for a series of pathlengths dl and integral is used:
I dI = - κ l [J] dl
∫I0 I ∫0
ln (I/I0) = - κ[J] l and since ln (I/I0) = ln 10 * log (I/I0)
Then - log (I/I0) = (1/2.303)κ[J] l = ε c l
This is describes the absorption at a single wavelength however sometimes the
absorption of an entire band is being treated and in this case the integrated
~ ~
absorption coefficient is used which corresponds to the area uinder the absorption
band Α =∫ε (v) dv
Bicomponent systems
• Two species present in solution
• Species behave independently
A = A1 + A2 = ε1c1l + ε2c2l = (ε1c1 + ε2c2)l
pH dependence of fluorescein

Isosbestic point
No change in abosrbance
Instrumentation

How do we record spectra?


Single beam diode array
spectrometer
Halogen lamp for visible
& deuterium lamp for UV

grating

Diode array detector


Single beam scanning
spectrometer
From Halogen lamp for visible
& deuterium lamp for UV Czerny-Turner monochromator

grating

To sample and then PMT Detector


Dual beam scanning
spectrometer
From Halogen lamp for visible
& deuterium lamp for UV

Two PMT (190-1100 nm)


or PbS (1100-2500 nm)
Detectors

From: http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/UV-Vis/spectrum.htm
Wavelength vs wavenumber
1.4 1.4

1.2

1.2
In converting from wavelength to
1.0
wavenumber you have to correct for
1.0
0.8 band pass. The slit width is constant in
nm over the whole spectrum but varies
Abs

0.6

in cm-1.
0.8 0.4
Abs

0.2

0.6
0.0
40000 35000 30000 25000 20000 15000
-1
wavenumber /cm
0.4

0.2

0.0
300 400 500 600 700 800
Wavelength /nm

5 nm, 1112 cm-1 5 nm, 156 cm-1


Sample Handling
• Spectroscopic grade solvents
• Concentration should be known
• Cuvette – generally quartz glass (QS), also OS or plastic
cuvettes can be used
• Typically 1 cm pathlength cuvettes (cells) are used
• Cleaning - hot nitric acid
• Cleaning – do not use of strong hydroxide bases or
sources of fluoride as these etch the glass surface
• UV/Vis light used to measure can also result in
photochemistry – do not expose the sample to light any
more than is necessary.
Measuring spectra
Single Beam vs. Dual beam spectrophotometers
Single beam (diode arrays) are fast but often less stable
First measure a solution not containing the compound of interest
(Spectrum 1)
Second measure a solution containing the compound of interest
(spectrum 2)
Subtract the reference spectrum from the spectrum with the sample
Dual beam (PMT) are slower but very stable
First measure a baseline using two solutions that do not contain the
compound of interest (baseline/background)
Second measure a solution containing the compound of interest
together with a blank solution
Subtract the reference spectrum from the spectrum with the sample
Measuring Molar Absorptivity
Measure absorbance over a range of
concentrations
Use different pathlength cells 1 mm to 10 cm
to increase dynamic range
Overlap data points
i.e. 1 mM with a 1 mm pathlength cell and
0.1 mM with a 1 cm cell
Deviations from linearity
• Assumption of independently absorbing species
– Inner filter effect
• Aggregation of samples at higher concentration
– Scattering
– Intermolecular interactions (H and J aggregates in perylenes)
• Photochemistry
• Change in gas content in solution
• Reaction with solvent
• Scatter – the wall!
Micellaneous topics
Photochromism
S

R
0.25
R
0.20
S

0.15 ∆

hv
Abs

0.10 Syn-folded
Anti-folded
S
0.05
R
0.00
R
250 300 350 400
S
Wavelength /nm
Pigments and Dyes II

1.0

0.8
S
S S

0.6
HH hv
H
-H2
H
Abs

0.4

0.2
H H
H H
hv -H2

250 300 350 400 450 500


Wavelength /nm
Solvatochroism
Interaction of a part of the chromophoric
group, e.g. a ketone, or amine, result in a
change in the energy of the frontier
orbitals and hence a shift in the absorption
spectrum
pH dependence of fluorescein

Isosbestic point
No change in abosrbance
Computational methods
N 2+
ZINDO/S 1 N
ZINDO/S H21 N
Ru
N _N N N
Me N N_ Me
N N N N
Ru
300 400 500 600 700 800 900
N N
N
TD-DFT 1
TD-DFT H21
= 2,2'-bipyridine
N N

300 400 500 600 700 800 900

MeCN 1
Predicted UV.Vis electronic
MeCN H21
spectra is done on the basis of
an isolated molecule (in
300 400 500 600 700 800 900
vacuo)
H2O 1
H2O H21
However solvent is not
innocent in determining the
absorptivity of chromophores
300 400 500 600 700 800 900
Band Gaps
Semiconductor materials: TiO2 (titanium dioxide –
white paint, some suncreams, solar cells), ITO
(Indium doped SnO2), NiO (nickel oxide)

CB (NiO)

Self cleaning windows generate active


hν oxygen and charges

VB (NiO)
Diffuse Reflectance

Uses an integrating sphere – used to look at rough surfaces


Also used for opaque solutions samples can be prepared as dilute
‘solutions’ in KBr
Specular reflectance

Brewster’s Angle
Polarisation angle (angle at which light
is fully polarised)
Applications: Following
Reactions
• Isosbestic points
• Multiple reaction steps S

R
0.25
R
0.20
S

0.15 ∆

hv
Abs

0.10 Syn-folded
Anti-folded
S
0.05
R
0.00
R
250 300 350 400
S
Wavelength /nm
Applications: Analysis
• Structural assignments
• Quantitative analysis
– HPLC
– Following reactions

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