Electronic Spectroscopy

Ultraviolet and visible

Where in the spectrum are these
transitions?
 Why should we learn this stuff?
After all, nobody solves structures
with UV any longer!

Many organic molecules have chromophores that absorb UV

UV absorbance is about 1000 x easier to detect per mole than NMR
Still used in following reactions where the chromophore changes. Useful
because timescale is so fast, and sensitivity so high. Kinetics, esp. in
biochemistry, enzymology.
Most quantitative Analytical chemistry in organic chemistry is conducted using
HPLC with UV detectors
One wavelength may not be the best for all compound in a mixture.
Affects quantitative interpretation of HPLC peak heights
 Uses for UV, continued
Knowing UV can help you know when to be skeptical of quant results. Need
to calibrate response factors
Assessing purity of a major peak in HPLC is improved by “diode array” data,
taking UV spectra at time points across a peak. Any differences could suggest
a unresolved component. “Peak Homogeneity” is key for purity analysis.
Sensitivity makes HPLC sensitive
e.g. validation of cleaning procedure for a production vessel
But you would need to know what compounds could and could not be detected
by UV detector! (Structure!!!)
One of the best ways for identifying the presence of acidic or basic groups, due
to big shifts in λ for a chromophore containing a phenol, carboxylic acid, etc.

“hypsochromic” shift “bathochromic” shift

λ
The UV Absorption process
•σ → σ * and σ → π * transitions: high-energy,
accessible in vacuum UV (λ max <150 nm). Not usually
observed in molecular UV-Vis.
•n → σ * and π → σ * transitions: non-bonding
electrons (lone pairs), wavelength (λ max ) in the 150-250
nm region.
•n → π * and π → π * transitions: most common
transitions observed in organic molecular UV-Vis,
observed in compounds with lone pairs and multiple
bonds with λ max = 200-600 nm.
•Any of these require that incoming photons match in
energy the gap corrresponding to a transition from
ground to excited state.
•Energies correspond to a 1-photon of 300 nm light are
 What are the π* π* π* Example
π* π* π* for a
π* π* π* simple
nature of these n n n
enone

absorptions?
π -π *; n-π *;
λ max =218 λ max =320
ε =11,000 ε =100
Example: π → π * transitions responsible for ethylene UV absorption at ~170 nm
calculated with ZINDO semi-empirical excited-states methods (Gaussian 03W):

hν 170nm photon

LUMO π antibonding molecular orbital

HOMO π u bonding molecular orbital g
 How Do UV spectrometers work?
Rotates, to
achieve scan
Two photomultiplier
inputs, differential
voltage drives amplifier.
Matched quartz cuvettes
Sample in solution at ca. 10-5 M.
System protects PM tube from
stray light
D2 lamp-UV
Tungsten lamp-Vis
Double Beam makes it a
difference technique
 Diode Array Detectors
Diode array
alternative puts
grating, array of
photosens.
Semiconductors after
the light goes through
the sample.
Advantage, speed,
sensitivity,
The Multiplex
advantage
Disadvantage,
Model from Agilent literature. Imagine resolution is 1 nm, vs
replacing “cell” with a microflow cell for 0.1 nm for normal
HPLC! UV
 Experimental details
What compounds show UV spectra?
Generally think of any unsaturated compounds as good
candidates. Conjugated double bonds are strong absorbers
Just heteroatoms are not enough but C=O are reliable
Most compounds have “end absorbance” at lower frequency. Unfortunately
solvent cutoffs preclude observation.
You will find molar absorbtivities ε in L•cm/mol, tabulated.
Transition metal complexes, inorganics
Solvent must be UV grade (great sensitivity to impurities with double
bonds)
The NIST databases have UV spectra for many compounds
An Electronic Spectrum Make solution of
concentration low enough
that A≤ 1
(Ensures Linear Beer’s
law behavior)
1.0
λ max with certain Even though a dual beam
goes through a solvent
extinction ε UV Visible blank, choose solvents
that are UV transparent.

Can extract the ε value

if conc. (M) and b (cm)
Absorbance

are known
UV bands are much
broader than the photonic
transition event. This is
because vibration levels
are superimposed on UV.

0.0
200 400 800
Wavelength, λ , generally in nanometers (nm)
 Solvents for UV (showing high
energy cutoffs)
Water 205 THF 220
CH3C≡ N 210 CH2Cl2 235

C6H12 210 CHCl3 245

Ether 210 CCl4 265

EtOH 210 benzene 280
Hexane 210 Acetone 300
MeOH 210 Various buffers for
HPLC, check before
Dioxane 220
using.
Organic compounds (many of
them) have UV spectra
One thing is clear
Uvs can be very non-specific
Its hard to interpret except at a
cursory level, and to say that the
spectrum is consistent with the
structure
Each band can be a
superposition of many
transitions
Generally we don’t assign the
particular transitions.
From Skoog and West et al. Ch 14
 An Example--Pulegone
Frequently
plotted as
log of
molar
extinction
ε
So at 240 nm,
pulegone has O

a molar
extinction of
7.24 x 103
Antilogof 3.86
 Can we calculate UVs?
MolarAbsorptivity (l/mol-cm) ElectronicSpectra
50243

40194

30146

20097

10049

nacindolA

0 Wavelength (nm)
220 230 240 250 260 270 280 290 300
M
olarAbsorptivity (l/m
ol-cm
) ElectronicSpectra
51972

41578

Semi-empirical (MOPAC) at 31183

AM1, then ZINDO for

config. interaction level 14 20789

10394
-1
Bandwidth set to 3200 cm
N
acetylindol

0 W
avelength (nm
220 230 240 250 260 270 280 290 300
 The orbitals involved

Molar Absorptivity (l/mol-cm) Electronic Spectra

55487

44390

Showing
33292
atoms whose
22195 MO’s
11097
contribute
Nacetylindolmost to the

0 Wavelength (nm) bands

200 210 220 230 240 250 260 270 280 290 300
 The Quantitative Picture
• Transmittance:
P0 P
T = P/P0 (power in) (power out)
• Absorbance:
A = -log10 T = log10 P0/P B(path through sample)

• The Beer-Lambert Law (a.k.a. Beer’s Law):

A = ε bc
Where the absorbance A has no units, since A = log10 P0 / P
ε is the molar absorbtivity with units of L mol-1 cm-1
b is the path length of the sample in cm
c is the concentration of the compound in solution, expressed in mol L-1 (or M, molarity)
 Beer-Lambert Law

Linear absorbance with increased concentration--directly

proportional
Makes UV useful for quantitative analysis and in HPLC
detectors
Above a certain concentration the linearity curves down,
loses direct proportionality--Due to molecular associations
at higher concentrations. Must demonstrate linearity in
validating response in an analytical procedure.
 Polyenes, and Unsaturated
Carbonyl groups;
an Empirical triumph
R.B. Woodward, L.F. Fieser and others
Predict λ max for π⇒π * in extended conjugation
systems to within ca. 2-3 nm.
Attached group increment, nm
Extend conjugation +30
Homoannular, base 253 nm
Addn exocyclic DB +5
Alkyl +5
Acyclic, base 217 nm O-Acyl 0
S-alkyl +30
O-alkyl +6

heteroannular, base 214 nm NR2 +60

Cl, Br +5
 Similar for Enones
β β O O

Ο
202 227 239
α 215
Base Values, add these increments…
X=H 207 α β γ δ ,+
x Extnd C=C +30
X=R 215
Add exocyclic C=C +5
X=OH 193
Homoannular diene +39
X=OR 193
alkyl +10 +12 +18 +18
With solvent correction OH +35 +30 +50
of…..
OAcyl +6 +6 +6 +6
Water +8
EtOH 0 O-alkyl +35 +30 +17 +31
CHCl3 -1 NR2
Dioxane -5 S-alkyl
Et2O -7 Cl/Br +15/+25 +12/+30
Hydrcrbn -11
Some Worked Examples
Base value 217
2 x alkyl subst. 10
exo DB 5
total 232
Obs. 237

Base value 214

3 x alkyl subst. 30
exo DB 5
total 234
Obs. 235

O Base value 215

2 ß alkyl subst. 24
total 239
Obs. 237
 Distinguish Isomers!
Base value 214
4 x alkyl subst. 20
exo DB 5
total 239
Obs. 238
HO2C

Base value 253

4 x alkyl subst. 20
total 273
Obs. 273

HO2C
Generally, extending conjugation
leads to red shift
“particle in a box” QM theory; bigger box
Substituents attached to a chromophore that cause a red shift
are called “auxochromes”
Strain has an effect…

λ max 253 239 256 248

 Interpretation of UV-Visible Spectra
• Transition metal
complexes; d, f
electrons.
• Lanthanide
complexes – sharp
lines caused by
“screening” of the f
electrons by other
orbitals
• One advantage of this
is the use of holmium
oxide filters (sharp
lines) for wavelength
calibration of UV
spectrometers. See Shriver et al. Inorganic Chemistry, 2
nd
Ed. Ch. 14
 Benzenoid Group K band (ε ) B band(ε ) R band

aromatics Alkyl
-OH
208(7800)
211(6200)
260(220)
270(1450)
--

-O- 236(9400) 287(2600)

UV of -OCH3 217(6400) 269(1500)

Benzene in NH2 230(8600) 280(1400)

heptane -F 204(6200) 254(900)

-Cl 210(7500) 257(170)
-Br 210(7500) 257(170)
-I 207(7000) 258/285(610/180)

-NH3+ 203(7500) 254(160)

-C=CH2 248(15000) 282(740)
-C≡ CH 248(17000) 278(6500
-C6H6 250(14000)
-C(=O)H 242(14000) 280(1400) 328(55)
-C(=O)R 238(13000) 276(800) 320(40)
-CO2H 226(9800) 272(850)
-CO2- 224(8700) 268(800)
From Crewes, Rodriguez, Jaspars, Organic Structure Analysis
-C≡ N 224(13000) 271(1000)
 Substituent effects don’t really add up
Can’t tell any thing about substitution geometry
Exception to this is when adjacent substituents
can interact, e.g hydrogen bonding.
E.g the secondary benzene band at 254 shifts to
303 in salicylic acid
In p-hydroxybenzoic acid, it is at the phenol or
benzoic acid frequency
 Heterocycles

Nitrogen heterocycles are pretty similar to the benzenoid

anaologs that are isoelectronic.
Can study protonation, complex formation (charge
transfer bands)
 Quantitative
analysis
Great for non-
aqueous titrations
Example here
gives detn of
endpoint for
Isosbestic points bromcresol green
Single clear point, can exclude
intermediate state, exclude light Binding studies
scattering and Beer’s law applies
Form I to form II

Binding of a lanthanide complex to

an oligonucleotide
 More Complex Electronic Processes
• Fluorescence: absorption of radiation
to an excited state, followed by
emission of radiation to a lower state
of the same multiplicity
• Phosphorescence: absorption of
radiation to an excited state, followed
by emission of radiation to a lower
state of different multiplicity
• Singlet state: spins are paired, no net
angular momentum (and no net
magnetic field)
• Triplet state: spins are unpaired, net
angular momentum (and net magnetic
field)