UV-Spectra

UV and Visible Spectroscopy

Vacuum UV or soft X-rays
100 - 200 nm
Quartz, O and CO absorb strongly in this region
2 2
N purge good down to 180 nm
2
Quartz region
200 – 350 nm
Source is D lamp
2
Visible region
350 – 800 nm
Source is tungsten lamp
Cells

UV Spectrophotometer
Quartz (crystalline silica)
 Visible Spectrophotometer
Glass
 IR Spectrophotometer
NaCl
Open-topped rectangular standard cell (a)
and apertured cell (b) for limited sample volume
Types of organic transitions
(Chromophores)
* •Sat’d hydrocarbons
•Vacuum UV

n* •Sat’d hydrocarbons with heteroatoms

•Possibly quartz UV

* •Olefins
•UV

n* •Olefins with heteroatoms

•UV
Examples of n* and  * transitions
Solvent effects
Hydrocarbons  water
*
Weak bathochromic or red shift
n*
Hypsochromic or blue shift (strongly affected by
hydrogen bonding solvents)
Solvent effects due to stabilization or
destabilization of ground or excited states,
changing the energy gap
UV cut-offs for common
solvents
Solvent choices
Important features to consider are solubility of sample
and UV cutoff of solvent
Filtration to remove particulates is useful to reduce
scattered light
Solvent purity is very important
Beer’s Law
I0
log10  A   lc
I
Io = Intensity of incident light
I = Intensity of transmitted light
 = molar extinction coefficient
l = path length of cell
c = concentration of sample
BEER LAMBERT LAW

Light
I0 I

Glass cell filled with

concentration of solution (C)

As the cell thickness increases, the intensity of I

(transmitted intensity of light ) decreases.
 
R- Transmittance
I
R= I0 - original light intensity
I0
I- transmitted light intensity
 
I
% Transmittance = 100 x
I0
1
Absorbance (A) or optical density (OD) = Log T
I0
= Log = 2 - Log%T
I
I
Log is proportional to C (concentration of solution) and is
I0
also proportional to L (length of light path
through the solution).
Relating Absorbance and
Transmittance
Absorbance rises linearly with
concentration. Absorbance is measured in
units.
Transmittance decreases in a non-linear
fashion.
Transmittance is measured as a %.
Absorbance = log10
(100/% transmittance)
A  CL = KCL by definition and it is called the
Beer Lambert Law.
A = KCL
K = Specific Extinction Coefficient ---- 1 g of
solute per liter of solution
 
A = ECL
E = Molar Extinction Coefficient ---- Extinction
Coefficient of a solution containing 1g molecule
of solute per 1 liter of solution
 Absorbance x Liter
E =
Moles x cm

E differs from K (Specific extinction Coefficient) by a

factor of molecular weight.

UNITS
  A = ECL
A = No unit (numerical number only)
Liter
E =
Cm x Mole
 L = Cm
C = Moles/Liter
Liter Mole
A = ECL = ( )x x Cm
Cm x Mole Liter

A = KCL
A = No unit C = Gram/Liter L = Cm
Liter
K=
Cm  Gram

Liter Gram
A = KLC = ( )x x Cm
Cm x Gram Liter
Every instrument has a useful range for a
particular analyte.
Often, you must determine that range
experimentally.
This is done by making a dilution series of
the known solution.  
These dilutions are used to make a working
curve.
Make a dilution series of a known quantity of
analyte and measure the Absorbance. Plot
concentrations v. Absorbance.
What concentration do you think the unknown
sample is?
In this graph, values above A=1.0 are not linear. If we use
readings above A=1.0, graph isn’t accurate.
The best range of this spectrophotometer is A=0.1 to
A=1.0, because of lower errors. A=0.4 is best.
Selected References
Harris, D. C., Bertolucci, M. D., Symmetry and
Spectroscopy, Dover, 1978.
Pasto, D. J., Johnson, C. R., Organic Structure
Determination, Prentice-Hall, 1969.
Drago, R. S., Physical Methods for Chemists,
Surfside Publishing, 1992.
Nakanishi, K., Berova, N., Woody, R. W., Circular
Dichroism, VCH Publishers, 1994
Williams, D. H., Fleming, I., Spectroscopic
methods in organic chemistry, McGraw-Hill, 1987.