UV Spectroscopy

Utility

• First organic spectral method; rarely used as a primary method for structure
determination

• Main contribution is that can readily identify the presence of conjugates π-systems
or unique chromophores

• Can sometimes be used to differentiate double bond isomers

• In combination with NMR and IR data can use to elucidate unique electronic
features not readily apparent from those methods

• Widely used in other applications

• Most common detector for HPLC

• Can be used to moniter reaction kinetics (chemistry, biology, medicine), etc.

 UV Spectroscopy
The Electromagnetic Spectrum

x-rays ultraviolet Infrared

(UV) visible microwaves radiowaves
(IR)

190 - 400 nm 400 - 800 nm

violet green orange

absorbed
color blue yellow red

observed
color
yellow red violet blue

short wavelength long wavelength

high frequency short frequency
high energy low energy
UV Spectroscopy
Colors of Different Wavelength Regions

Absorbance & Transmittance

wavelength absorbed transmitted color

absorbed (nm) color (compliment)

< 380 ultraviolet ---

380 - 435 violet yellowish green
435 - 480 blue yellow
480 - 490 greenish blue orange
490 - 500 bluish green red
500 - 560 green purple
560 - 580 yellowish green violet
580 - 590 yellow blue
590 - 625 orange greenish blue
625 - 780 red bluish green
> 780 near IR red
UV Spectroscopy
Origin of the Absorption

• The absorption of UV or visible radiation corresponds to the excitation of valence

electrons

• Valence electrons are typically found in:

- σ bonding orbitals (single bonds)
- π bonding prbitals (double or triple bonds)
- non-bonding orbitals (lone pair electrons)

LUMO

ΔE = [Eexcited – Eground] = hν

HOMO
 σ σ* alkanes
UV Spectroscopy
σ π* carbonyls
Electronic Transitions
π π* alkenes, carbonyls, alkynes, etc.

n σ* heteroatoms (O, N, S, X, etc.)

ΔE = [Eexcited – Eground] = hν

n π* carbonyls

orbitals possible electronic transitions

σ*

antibonding

π*

E E
atomic n

bonding
σ
UV Spectroscopy
Electronic Transitions

Relative Transition Energies

σ σ* > n σ* > π π* > n π* > σ π*

most useful

• not all transformations that are possible will be observed

• some electronic transitions “forbidden” by certain selection rules
• even forbidden transitions can be observed, but usually not very intense
for example, n  π*
UV Spectroscopy
The Spectrometer
UV Spectroscopy
Beer-Lambert Law

describes relationship between absorbance

and concentration

ℓ
A = log ( I0 / I ) = ε ℓ c

Where:
A is absorbance (no units)
I0 = intensity of incident light
I = intensity transmitted light
ε = molar absorbtivity or extinction coefficient
ℓ = path length; length of the sample cell (cm)
c = sample concentration (mol/L)
UV Spectroscopy
Spectrum Features

isoprene

• values of 104-106 are termed high intensity absorptions

ε=A/ℓc • values of 103-104 are termed low intensity absorptions
• values of 0 to 103 are the absorptions of forbidden transitions
UV Spectroscopy
Spectrum Features

reporting data
NH2

O
O
λmax = 206 nm
252
317
376
UV Spectroscopy
Spectrum Features

peak broadening
UV Spectroscopy
Choice of Solvent

Solvent should not absorb UV radiation in the same region as the sample
(measuring UV spectra below 200 nm is impractical)

solvent cutoff (nm) solvent cutoff (nm)

acetonitrile 190 ethanol 205
chloroform 240 hexane 201
cyclohexane 195 methanol 205
diethyl ether 210 isooctane 195
dioxane 215 water 190

sample cell: quartz glass cutoff 210 nm

UV Spectroscopy
Solvent Effect on Spectra

OH
UV Spectroscopy
Chromophores

Basic Information

• All molecules capable of absorbing ultraviolet radiation, though most do so at very

high energy (wavelengths < 200 nm).
• Unsaturated groups which give rise to absorptions involving π or π* orbitals in the
in the near-UV/visible region are called chromophores.
• Saturated groups with non-bonding electrons, which can give rise to transitions
involving non-bonding orbitals are called auxochromes.
• Most useful transitions for analysis are the intense π → π * transitions and the
weaker, but lower energy, n → π* transitions.
UV Spectroscopy
Chromophores

Absorptions of Organic Molecules

Alkanes:
• Saturated molecules that lack lone pairs
• Only transitions possible are σ  σ*
• high energy; absorb UV radiation at very short wavelengths
• not accessible using UV spectroscopy

Alcohols, Ethers, Amines, & Sulfur Compounds:

• Satuated molecules with lone pairs of electrons
• Important transitions are n  σ*
• high energy, most often at wavelengths shorter than 200 nm
• alcohols and amines: 175 – 200 nm
• thiols and sulfides: 200 – 220 nm
UV Spectroscopy
Chromophores

Absorptions of Organic Molecules

Alkenes & Alkynes:

• Important transitions are π  π*
• high energy, but impacted by substitution
• simple alkenes: 175 nm
• simple alkynes: 170 nm

Carbonyls:
• Important transitions are π  π* (188 nm)
• n  π* also possible (280 – 290 nm)
• sensitive to substitution
UV Spectroscopy
Chromophores
UV Spectroscopy
Chromophores

Terminology for Absorptive Shifts

Bathochromic shift: shift of absorption to a longer wavelength

Hypsochromic shift: shift of absorption to a shorter wavelength
Hyperchromic effect: an increase in absorption intensity
Hypochromic effect: a decrease in absorption intensity

Hyperchromic

ε

Hypsochromic Bathochromic
Hypochromic

200 nm 700 nm
UV Spectroscopy
Conjugation Effects
compound λmax (nm) ε

H2C CH2 175 15,000

217 21,000

258 35,000

465 125,000

O n π* 280 12
π π* 189 900

O
n π* 280 27
π π* 213 7,100
UV Spectroscopy
Conjugation Effects

• Greater the conjugation, the lower

the energy required to induce
electronic transitions (e.g. the
(longer the wavelength)

• lengthening conjugation also

increases band intensity
(greater molar absorbitivity)

• adding substitutents may have

same effect, but to a much
smaller degree
UV Spectroscopy
Conjugation Effects

Extended π Systems

H2C CH2

π*

Energy

π

ethylene

butadiene
hexatriene
octatetraene
UV Spectroscopy
Auxochromes & Conjugation

• atoms with lone pairs can extend conjugation by resonance

X X
R R

X = OH, OR, NH2, halogen, etc.

• alkyl substituents can influence wavelength by overlap of C-H bonding orbital

with the π system (e.g. by hyperconjugation)
UV Spectroscopy
Woodward Fieser Rules for Dienes

acyclic dienes cyclic dienes

s-trans s-cis homoannular heteroannular

(cisoid) (transoid)

• Woodward & Fieser derived a set of empirical rules for the estimation of wavelength for
the low energy π  π*electronic transition

• Based on empirical observation of known conjugated structures

• Can be used to reliably predict absorption wavelength in dienes, enone, and to a lesser
extent aromatic systems
UV Spectroscopy
Woodward Fieser Rules for Dienes

s-trans homoannular heteroannular

(cisoid) (transoid)

base values: 217 nm 253 nm 214 nm

Increments:

For each additional conjugated double bond + 30 nm

For each exocyclic double bond + 5 nm
For each alkyl group + 5 nm
For each of the following groups:
- OR + 6 nm
- O(C=O)R + 0 nm
- Cl + 5 nm
- Br + 5 nm
not affected by solvent
- SR + 30 nm
- NR2 + 60 nm
- Ph + 60 nm

Where both types of cyclic dienes are present, the base with the longer λmax is used.
UV Spectroscopy
Woodward Fieser Rules for Dienes

CAUTION!

R
This compound had three exocyclic
double bonds; the indicated bond is
exocyclic to two rings.
λmax calc = 284

this is not a heteroannular diene; must

use base value for acyclic diene.
λmax calc = 232

this is not a homoannular diene; must

use base value for acyclic diene
λmax calc = 237
UV Spectroscopy
Woodward Fieser Rules for Dienes

examples 217 nm
acyclic diene
3 alkyl subst 15 nm
calculated value 232 nm
observed 234 nm

acyclic diene 217 nm

2 alkyl subst 10 nm
1 exocyclic db 5 nm
calculated value 232 nm
observed 236 nm

cisoid diene 253 nm

4 alkyl subst 20 nm
1 exocyclic db 5 nm
calculated value 278 nm
observed 275 nm
UV Spectroscopy
Woodward Fieser Rules for Dienes

examples

transoid diene 214 nm

3 alkyl subst 15 nm
EtO 1 OR subst 6 nm
1 exocyclic db 5 nm
calculated value 240 nm
observed 241 nm

cisoid diene 253 nm

2 conj db 60 nm
5 alkyl subst 25 nm
O 1 acyl subst 0 nm
3 exocyclic db 15 nm
O
calculated value 353 nm
observed 355 nm
UV Spectroscopy
Woodward Fieser Rules for Dienes

double bond regioisomers

OH OH
O O

abietic acid levopimaric acid

λmax calc: 239 λmax calc: 278

λmax obs: 238 λmax obs: 275

transoid diene 214 nm cisoid diene 253 nm

4 alkyl subst 20 nm 4 alkyl subst 20 nm
1 exocyclic db 5 nm 1 exocyclic db 5 nm
calculated value 239 nm calculated value 278 nm
UV Spectroscopy
Fieser-Kuhn Rules for Extended Polyenes

• Woodward-Fieser Rules work well up to four conjugated double bonds

• For more extended conjugation, use the Fieser-Kuhn Rules

λmax = 114 + 5(# alkyl substituents) = n(48 - 1.7n) - 16.5(# endo) - 10(# exo)

where n = number of conjugated double bonds

lycopene

λmax = 114 + 5(8) = 11(48 - 1.7•11) - 16.5(0) - 10(0) = 476 nm

observed: 474 nm

β-carotene

λmax = 114 + 5(10) = 11(48 - 1.7•11) - 16.5(2) - 10(0) = 453 nm

observed: 452 nm
 Solvent Correction nm
H2O + 8
EtOH 0
UV Spectroscopy CHCl3
Dioxane
- 1
- 5
Et2O - 7
Woodward Fieser Rules for Enones Hydrocarbon - 11

O O
O O

R R

acyclic enone 6-membered ring 5-membered ring acyclic dienone

enone enone

base values: 215 nm 215 nm 202 nm 245 nm

Increments: α β γ δ and higher

For each additional conjugated double bond + 30 nm

For each exocyclic double bond + 5 nm
For each homodiene component + 39 nm

For each alkyl group + 10 nm + 12 nm + 18 nm + 18 nm

For each of the following groups: - OH + 35 nm + 30 nm + 50 nm
- OR + 35 nm + 30 nm + 17 nm + 31 nm
- O(C=O)R + 6 nm + 6 nm + 6 nm + 6 nm
- Cl + 15 nm + 12 nm
- Br + 25 nm + 30 nm
β δ β - SR + 30 nm + 85 nm
O O
- NR2 + 60 nm + 95 nm
β R R
α γ α
UV Spectroscopy
Woodward Fieser Rules for Enones

examples

O acyclic enone 215 nm

1 α alkyl 10
2 β alkyl 24 nm
calculated value 249 nm
observed 249 nm

5-membere enone 202 nm

1 α Br 25 nm
O 2 β alkyl 24 nm
1 exocyclic db 5 nm
Br
calculated value 256 nm
observed 251 nm
UV Spectroscopy
Woodward Fieser Rules for Enones

practice

Me Me Me
Br2 Br
or
O O O
H H H
Br

base base

Me Me
or
O O
H

Can you distinguish the two by UV?

UV Spectroscopy
Woodward Fieser Rules for Enones

practice

Me Me
vs
O O
H

6-membered enone 215 nm 215 nm

β alkyl 12 nm 24 nm
exocyclic db 5 nm
227 nm 244 nm
UV Spectroscopy
Woodward Fieser Rules for Enones

practice

absorbance in EtOH?
UV Spectroscopy
Woodward Fieser Rules for Other Conjugated Carbonyls

β β
O O

β H β OR

α α
(R = H or R')

aldehydes carboxylic acid or ester

Base Values

unsubstituted aldehyde or ester 208 nm 193 nm (not observed)

with α or β alkyl groups 220 nm 208 nm
with α,β or β,β alkyl groups 230 nm 217 nm
with α,β,β alkyl groups 242 nm 225 nm

for an exocyclic α,β double bond + 5 nm

for an endocyclic α,β double bond + 5 nm
in a 5- or 7-membered ring

as for enones, solvent correction is also relevant

UV Spectroscopy
Woodward Fieser Rules for Aromatic Compounds

• Substitution, auxochromic groups, conjugation and solvent effects can cause

shifts in wavelength and intensity of bands for aromatic systems similar to
dienes and enones

• Show multiple bands, often fine structure

• However, shifts are difficult to predict – the formulation of empirical rules is often
dificult (there are more exceptions than rules)

• Can make some useful predictions for benzoyl derivatives

We will not worry about predicting UV bands

in aromatic compounds
UV Spectroscopy
Woodward Fieser Rules for Benzoyl Derivatives

O O O

R H OR

aryl ketones benzaldehydes benzoic acids

(R = alkyl) and esters

base values: 246 nm 250 nm 230 nm

Increments: ortho meta para

For each alkyl group + 3 nm + 3 nm + 10 nm

For each OH or OR (R = alkyl) + 7 nm + 7 nm + 25 nm
For each O + 11 nm + 20 nm + 78 nm
For each of the following groups:
- Cl + 0 nm + 0 nm + 10 nm
- Br + 2 nm + 2 nm + 15 nm
- NH2 + 13 nm + 13 nm + 58 nm
- NH(C=O)CH3 + 20 nm + 20 nm + 45 nm
- NHCH3 + 73 nm
- N(CH3)2 + 20 nm + 20 nm + 85 nm