UV-VIS CHM 3402

Spectroscopy Dr. Mohd Rashidi Abdull Manap

 1-Molecular Electronic Transition
Learning in Electronic Spectroscopy
Outcomes
Students 2-Instrumentation/Uv-Vis
should be able Spectrometer
to understand:
3-Application and Interpretation
of a UV-Vis spectrum
1-Molecular Electronic Transition in
Electronic Spectroscopy

[Link]
/1505050/Organic/Arenes/Chapter%205/sec5-14/[Link]
 When red photons are
absorbed from white
lights, your eyes see green
color

[Link]
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[Link]
oPhotometer_for_authenticity_testing_of_spirits.pdf
[Link]
net/figure/UV-VIS-spectra-
of-diverse-carotenoids-imp
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fitness/refined-oil-vs-filtered-oil-
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•Experimental UV–Visible
absorption peaks arising due to
electronic transitions
•This is due to the presence of
chemical species in the sample
•The electronic transitions can be
understand by the means of
molecular orbitals
[Link]
tion-spectrophotometry
The valence electrons are the only ones whose energies permit them to be
excited by near UV/visible radiation.
s* (anti-bonding)

p* (anti-bonding)
Four types of transitions
*
n (non-bonding) *

n*
p (bonding)
n*

s (bonding)
s  s* transition in vacuum UV
n  s* saturated compounds with non-bonding electrons
n ~ 150-250 nm
e ~ 100-3000 ( not strong)
n  p*, p  p* requires unsaturated functional groups (eq. double bonds)
most commonly used, energy good range for UV/Vis
n ~ 200 - 700 nm
n  p* : e ~ 10-100
p  p*: e ~ 1000 – 10,000
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Chromophore Excitation lmax, nm Solvent

C=C p→p* 171 hexane

C=O
n→p* 290 hexane
p→p* 180 hexane

N=O
n→p* 275 ethanol
p→p* 200 ethanol

C-X   n→s* 205 hexane

X=Br, I n→s* 255 hexane
UV Spectrum of Isoprene

=>
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 s* s*
C C p* p*
hv
= hv
=hc/
p p
s s
 
Example: ethylene absorbs at max = 165 nm = 10,000

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C O
s* s*
p* p*
n hv
n

p p
s s
n 
The n to p* transition is at even longer wavelengths but is not as strong as p to p*
transitions. It is said to be “forbidden.”
Example:
Acetone: n max = 188 nm ; = 1860
n max = 279 nm ; = 15

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 Conjugated systems:

C C

LUMO

HOMO

• Preferred transition is between Highest Occupied Molecular Orbital

(HOMO) and Lowest Unoccupied Molecular Orbital (LUMO).
• Additional conjugation (double bonds) lowers the
HOMO-LUMO energy gap. 21
Similar structures have similar UV spectra:

O
O

max = 238, 305 nm max = 240, 311 nm max = 173, 192 nm

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Absorption Characteristics of Some Common Chromophores
Chromophore Example Solvent lmax (nm) emax Type of
transition
Alkene C6H13HC CH2 n-Heptane 177 13,000 pp*
Alkyne n-Heptane 178 10,000 pp*
C5H11C C CH3 196 2,000
_
225 160
_
Carbonyl O n-Hexane 186 1,000 ns*
280 16
np*
CH3CCH3
n-Hexane 180 Large
O ns*
293 12
CH3CH np*
Carboxyl O Ethanol 204 41 np*
CH3COH
Amido O Water 214 60 np*
CH3CNH2
Azo H3CN NCH3 Ethanol 339 5 np*
Nitro CH3NO2 Isooctane 280 22 np*
Nitroso C4H9NO Ethyl ether 300 100 _
665 20 np*
Nitrate C2H5ONO2 Dioxane 270 12 np* 23
Structure elucidation of organic
compounds
UV-Vis spectroscopy is useful in the
structure elucidation
• The presence or absence of unsaturation
• The presence of hetero atoms

Refer the wavelength position or

combination of peaks
What UV-Vis can observe?

Conjugated systems (alternating single

and double bond)

Metal-Ligand interaction in transition metal

complexex
UV-Vis Spectrometer

It measures absorbance of UV or Light

by a sample (solid, liquid, gas)

Measurement can take place in

single/multiple wavelength or in range
(spectral region)
 2-Instrumentation/Uv-Vis
Spectrometer
Coherence Grating Cuvettes (aq or (Qualitative and
UV- System organic solution) Quantitative)
Deuterium
Lamps and
Vis-Tungten
Lamp
sources-

Beer’s Law Absorption,

Transmission,
Reflectivity of Materials
UV-Vis
Spectrometer-
liquid sample
UV-Vis Spectrometer-solid sample
(crystals, foods etc)
Portable UV-Vis Spectrometer

[Link]
000#/840-300000
UV-Vis Spectroscopic Samples
Electronic Spectra

[Link]
3-Application and Interpretation of a
UV-Vis spectrum
• How does UV-Vis method fit into this process?

UV-Vis
Wavelength(nm) Functional Groups

Shift in Molecular
wavelength environment
 UV-Vis Spectra (300 to 900 nm) 𝝀𝒎𝒂𝒙 =? 𝒏𝒎
Beer’s Law in
UV-Vis
Spectroscopy

e.g Effect of path

lengths

[Link]
Quantitative Analysis with UV-Vis
Beer’s Law
A=ebc
e: the molar absorptivity (L mol-1 cm-1)
b: the path length of the sample
c :the concentration of the compound in
solution, expressed in mol L-1
 Path
length / cm 0 0.2 0.4 0.6 0.8 1.0

%T 100 50 25 12.5 6.25 3.125

Absorbance
External Standard and the Calibration Curve
Standard Addition Method
• Standard addition must be used whenever the matrix of
a sample changes the analytical sensitivity of the
method. In other words, the slope of the working curve
for standards made with distilled water is different
from the same working curve.
 Prepare the Standards

The concentration and volume of the stock

solution
added should be chosen to increase the
concentration of the unknown by about 30% in
each succeeding flask.
A  bC
bVx C x bVs Cs
A   kVx C x  kVs Cs
Vt Vt
y  b  ax (a  kVs , x  C s , b  kVx C x )
Vs Cs
A  0  kVx C x  kVs C s  C x  
Vx

Cx: unknown concentration

Molecular structure determination with
UV-Vis
C11H7ClN2O2S in EtOH

376, 334, 262 and 246 nm

no absorption peak above 400 nm

376 nm n→π* transition azomethine 

332 nm π→π* transition of thiophene and benzene

[Link]
Molecular structure determination with
UV-Vis

Blue spectrum
360 nm - π−π*  transition
530 nm - n−π* transition
[Link]
Molecular structure determination with
UV-Vis

Caffeine in DCM
Below 243 nm -solvent
274.7 nm n→π* transition of lone pair in caffeine C=O
[Link]
[Link]
[Link]
Factors affecting absorption
• The solvent

• Degree of conjugation

• Acid-base indicators (pH)

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 UV Spectral Nomenclature

Blue Shift (Hypsochromic) – Red Shift (Bathochromic) –

Peaks shift to shorter wavelength. Peaks shift to longer wavelength.

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Red line shows the absorption peak intensities of caffeine are
reduced and blue shifted (360 nm) and a new (287 nm) peak is
formed. 
[Link]
 - For Compounds with Multiple Chromophores:

If isolated (more than one single bond apart)

- e are additive
- l constant

CH3CH2CH2CH=CH2 lmax= 184 emax = ~10,000

CH2=CHCH2CH2CH=CH2 lmax=185 emax = ~20,000

If conjugated - shifts to higher l (red shift)

1,3 butadiene: lmax= 217 nm ; emax= 21,000

1,3,5-hexatriene lmax= 258 nm ; emax= 35,000

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- For Compounds with Multiple Chromophores:

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 Solvent Effects - Intensity
Solvents can induce significant changes in the intensity of peaks.

Hyperchromic – Increase in absorption intensity.

Hypochromic – Decrease in absorption intensity.

Absorption characteristics of 2-methylpyridine

Solvent max max

Hexane 260 2000
Chloroform 263 4500
Ethanol 260 4000
Water 260 4000
Ethanol - HCl (1:1) 262 5200 55
 Solvent effects
• The excited states of most π→π* transitions are more
polar than their ground states because a greater charge
separation is observed in the excited state.

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• If a polar solvent is used the dipole–dipole interaction reduces the energy
of the excited state more than the ground state, hence the absorption in a
polar solvent such as ethanol will be at a longer wavelength (lower energy,
hence lower frequency) than in a non-polar solvent such as hexane.

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 Solvent effects
• For n -> π* transition, the n state is much more easily
stabilized by polar solvent effects (H-bonds and
association), so in going from nonpolar to polar solvent
there is a blue shift or hypsochromic shift (decrease in
λmax, increase in ΔE).

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 O

n* Hypsochromic shift

methanol heptane

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Solvent Effects

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Thank you