Lecture 5

CHEM F313

UV-Vis Spectroscopy
30-10-21
CHROMOPHORE:
• Originally, term chromophore was applied to the system responsible for imparting color to
a compound.
• However, many groups absorb light but do not impart color
• Chromophore, is thus defined as any functional group that absorbs electromagnetic
radiation.

Chromophore Example Excitation λmax, nm ε Solvent

C=C Ethene π __> π* 171 15,000 hexane

C≡C 1-Hexyne π __> π* 180 10,000 hexane
n __> π* 290 15 hexane
C=O Acetone
π __> π* 180 10,000
n __> π* 275 17 ethanol
N=O Nitromethane
π __> π* 200 5,000

C-X, X=Br Methyl bromide; n __> σ* 205 200 hexane

X=I Methyl Iodide n __> σ* 255 360
AUXOCHROME:

• Originally defined as: A group that could enhance the color imparting properties of a
chromophore without being itself a chromophore. Example: -OR, -NH2, -NR2, etc.

• Auxochromes have a synergist effect that is coupled with their ability to extend the
conjugation of a chromophore.

• Thus, in true sense, auxochrome becomes a part of chromophore by extending

conjugation. In other words it adds a chromophore to a chromophore.

• Auxochrome can be a saturated or unsaturated group with non-bonding electrons that

helps in extending the conjugation of a chromophore.

• Auxochrome impacts both wavelength and intensity of absorption.

EXAMPLES:
 Example from Nature: Green Color of Chlorophylls
VIBGYOR
Violet: 400 - 420 nm
Indigo: 420 - 440 nm
Blue: 440 - 490 nm
Green: 490 - 570 nm
Yellow: 570 - 585 nm
Orange: 585 - 620 nm
Red: 620 - 780 nm
 UV-Visible Spectroscopy: Key Points
❖Most Quantitative Analytical Chemistry in organic/inorganic
chemistry/Biological chemistry is conducted using UV-visible studies.
❖Many organic molecules have chromophores that absorb UV.
❖Easy and relatively simple technique
❖Used in reactions where the chromophore changes. Useful because timescale
is so fast, and sensitivity so high. Kinetics, especially in biochemistry and
enzymology.
❖One wavelength may not be the best for all compound in a mixture.
❖Used in quantitative interpretation of HPLC peak heights.
The wavelength and amount of light that a compound absorbs depends on its
molecular structure and the concentration of the compound used.

The concentration dependence follows Beer’s Law.

A = εlc = log I0/I
Where A is absorbance
ε is the molar absorptivity with units of L mol-1 cm-1
l is the path length of the sample (typically in cm).
c is the concentration of the compound in solution, expressed in mol L-1
 Isoprene in hexane solution

A = 0.82 at 222 nm for 4x10 -5 molar solution when 1 cm path length cuvette
was used.  = ?
 UV spectrometers
Rotates, to achieve scan
Matched quartz cuvettes
Sample in solution at ca. 10-5 M.
D2 lamp-UV
Tungsten lamp-Vis
Double Beam makes it a different technique

Two photomultiplier inputs,

differential voltage drives amplifier.
 Chromophore λmax (nm) εmax
(L mol-1cm-1)
C=C 175 14,000
C≡C 175 10,000
195 2,000
223 150
C=O 160 18,000
185 5,000
280 15
R-NO2 200 5,000
274 15
C≡N 165 5
C=C─C=C 217 20,000
C=C─C=O 220 10,000
315 30
C=C─C≡C 220 7,500
230 7,500
Benzene 184 60,000
204 7,400
255 204
Types of electrons available for electronic
transitions:

σ, π, n electrons
 1. Sigma (s) – single bond electron

Low energy bonding orbital High energy anti-bonding orbital

- Single bonds are usually too high in excitation energy for most instruments (185 nm)

Vacuum UV: Most compounds of atmosphere absorb in this range, so difficult to work
with.
 2. Pi (p) – double bond electron

Low energy bonding orbital High energy anti-bonding orbital

3. Non-bonding electrons (n): don’t take part in

any bond, neutral energy level.
Example: Formaldehyde
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
s→s*
n (non-bonding)
n→s*

p→p*
p (bonding)
n→p*

s (bonding)

σ → σ* transition in vacuum UV
n → σ* saturated compounds with non-bonding electrons
λ  150-250 nm
  100-3000 M-1cm-1 ( not strong)
n → π*, π → π* requires unsaturated functional groups (eq. double bonds) most commonly used, energy good range for UV/Vis
λ  200 - 700 nm
n → π* :   10-100 M-1cm-1
π → π*:   1000 – 10,000 M-1cm-1
 n→s* Transitions
High in energy.  between 150 and 250 nm.
Not many molecules with n→s* transitions in UV/vis region
 max max
H2O 167 1480
CH3OH 184 150
CH3Cl 173 200
CH3I 258 365
(CH3)2S 229 140
(CH3)2O 184 2520
CH3NH2 215 600
(CH3)3N 227 900
 n→p* and p→p* Transitions
❖Most UV/vis spectra involve these transitions.
❖p→p* are generally more intense than n→p*.
 max  max type
C6H13CH=CH2 177 13000 p→p*
C5H11CC–CH3 178 10000 p→p*
O
CH3CCH3 186,275, 150 1000 n→s*, n→p*, p→p*
O
CH3COH 204 41 n→p*
CH3NO2 280 22 n→p*
CH3N=NCH3 339 5 n→p*
Transition Probability: It is not essential that a compound on exposure to ultraviolet or
visible light must always gives rise to an electronic transition. However, the probability of a
particular electronic transition has been found to depend on , the value of molar extinction
coefficient and certain other factors. Accordingly, transitions can be divided into two
categories.

(i) Allowed transitions – these are transitions having molar coefficient 104 or more.
These are generally designated as p→ p* transitions. For example in 1,3-butadiene
which exhibits absorption at 217 nm has max value 21000 represents an allowed
transition. These transition are mainly favored due to symmetry relationship.

(ii) Forbidden transitions – these are transitions for which max is generally less than
104 . for example transition of saturated aldehyde showing weak absorption near 290 nm
and having max 100 has been a forbidden transition. Ex: Carbonyl group absorbs at 300 nm
and a molar absorptivity of 10-100.
 ALKANES: σ → σ* Requires short λ 150 nm
s* s*
C C p* p*

hv = hv
=hc/
p p
s s
p p*
Example: Ethylene
Several transitions are available but lowest energy transition is
the most important. 170 to 190 nm in non conjugated alkene
Saturated Aliphatic ketones
s* s*
C O p* p*
n hv n

p p
s s
An electron of unshared electron pair on a hetero atom is excited to π* antibonding orbital
Lowest energy is n p* But “forbidden” due to Symmetry consideration low intensity even
though long wavelength
Other two transitions are n→σ* and π→ π *
The n to p* transition appears at even longer wavelengths but is not as strong as p to p* transitions. It is
said to be “forbidden” transition.
Example:
Acetone: n−s*  max = 188 nm ;  = 1860 M-1cm-1
n−p*  max = 279 nm ;  = 15 M-1cm-1
p−p*  max = 195 nm (more intense)
C C s − s* 135 nm

C C p − p* 165 nm

H
C O n − s* 183 nm weak

p − p* 150 nm
C O n − s* 188 nm
n − p* 279 nm weak

188 nm

C O A
279 nm


Absorption Characteristics of Some Common Chromophores
Chromophore Example Solvent  max (nm)  max Type of
transition
Alkene C6H13HC CH2 n-Heptane 177 13,000 p→p*
Alkyne n-Heptane 178 10,000 p→p*
C5H 11C C CH3 196 2,000 _
225 160 _
Carbonyl O n-Hexane 186 1,000 n→s*
280 16 n→p*
CH3CCH3
O n-Hexane 180 ~1,000 n→s*
CH3CH 293 12 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*