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UV-Visible Spectroscopy

A diagram of the components of a typical spectrometer are shown in the following diagram. The functioning
of this instrument is relatively straightforward. A beam of light from a visible and/or UV light source (colored
red) is separated into its component wavelengths by a prism or diffraction grating. Each monochromatic
(single wavelength) beam in turn is split into two equal intensity beams by a half-mirrored device. One
beam, the sample beam (colored magenta), passes through a small transparent container (cuvette)
containing a solution of the compound being studied in a transparent solvent. The other beam, the
reference (colored blue), passes through an identical cuvette containing only the solvent. The intensities of
these light beams are then measured by electronic detectors and compared. The intensity of the reference
beam, which should have suffered little or no light absorption, is defined as I0. The intensity of the sample
beam is defined as I. Over a short period of time, the spectrometer automatically scans all the component
wavelengths in the manner described. The ultraviolet (UV) region scanned is normally from 200 to 400 nm,
and the visible portion is from 400 to 800 nm.

If the sample compound does not absorb light of of a given
wavelength, I = I
0
. However, if the sample compound absorbs light
then I is less than I
0
, and this difference may be plotted on a graph versus
wavelength, as shown on the right. Absorption may be presented
as transmittance (T = I/I
0
) or absorbance (A= log I
0
/I). If no absorption
has occurred, T = 1.0 and A= 0. Most spectrometers display absorbance
on the vertical axis, and the commonly observed range is from 0 (100%
transmittance) to 2 (1% transmittance). The wavelength of maximum
absorbance is a characteristic value, designated as
max
.
Different compounds may have very different absorption maxima and
absorbances. Intensely absorbing compounds must be examined in dilute
solution, so that significant light energy is received by the detector, and
this requires the use of completely transparent (non-absorbing) solvents.
The most commonly used solvents are water, ethanol, hexane and
cyclohexane. Solvents having double or triple bonds, or heavy atoms (e.g.
S, Br & I) are generally avoided. Because the absorbance of a sample will
be proportional to its molar concentration in the sample cuvette, a
corrected absorption value known as the molar absorptivity is used when
comparing the spectra of different compounds. This is defined as:
Molar Absorptivity,
= A/ c l
( where A= absorbance, c = sample
concentration in moles/liter
& l = length of light path through the
cuvette in cm.)



For the spectrum on the right, a solution of 0.249 mg of the unsaturated aldehyde in 95% ethanol
(1.42 10
-5
M) was placed in a 1 cm cuvette for measurement. Using the above formula, = 36,600 for the
395 nm peak, and 14,000 for the 255 nm peak. Note that the absorption extends into the visible region of
the spectrum, so it is not surprising that this compound is orange colored.
Molar absorptivities may be very large for strongly absorbing compounds ( >10,000) and very small if
absorption is weak ( = 10 to 100).




End of this supplementary topic

Empirical Rules for Absorption Wavelengths of Conjugated Systems
Woodward-Fieser Rules for Calculating the
max
of Conjugated Dienes and Polyenes
Core Chromophore Substituent and Influence

Transoid Diene
215 nm
R- (Alkyl Group) .... +5 nm
RO- (Alkoxy Group) .. +6
X- (Cl- or Br-) ......... +10
RCO
2
- (Acyl Group) .... 0
RS- (Sulfide Group) .. +30
R
2
N- (Amino Group) .. +60
Further -Conjugation
C=C (Double Bond) ... +30
C
6
H
5
(Phenyl Group) ... +60

Cyclohexadiene*
260 nm
(i) Each exocyclic double bond adds 5 nm. In the example on the right, there are
two exo-double bond components: one to ring A and the other to ring B.
(ii) Solvent effects are minor.
* When a homoannular (same ring) cyclohexadiene chromophore is present, a base value of
260 nm should be chosen. This includes the ring substituents. Rings of other size have a lesser
influence.
max (calculated) = Base (215 or 260) + Substituent Contributions

Some examples that illustrate these rules follow.




Woodward-Fieser Rules for Calculating the
__
> *
max
of Conjugated Carbonyl Compounds
Core Chromophore Substituent and Influence
R = Alkyl 215 nm
R = H 210 nm
R = OR' 195 nm
- Substituent
R- (Alkyl Group) +10 nm
Cl- (Chloro Group) +15
Br- (Chloro Group) +25
HO- (Hydroxyl Group) +35
RO- (Alkoxyl Group) +35
RCO
2
- (Acyl Group) +6
- Substituent
R- (Alkyl Group) +12 nm
Cl- (Chloro Group) +12

Cyclopentenone
202 nm

Br- (Chloro Group) +30
HO- (Hydroxyl Group) +30
RO- (Alkoxyl Group) +30
RCO
2
- (Acyl Group) +6
RS- (Sulfide Group) +85
R
2
N- (Amino Group) +95
& - Substituents
R- (Alkyl Group) +18 nm (both & )
HO- (Hydroxyl Group) +50 nm ()
RO- (Alkoxyl Group) +30 nm ()
Further -Conjugation
C=C (Double Bond) ... +30
C
6
H
5
(Phenyl Group) ... +60
(i) Each exocyclic double bond adds 5 nm. In the example on the right, there are two exo-
double bond components: one to ring A and the other to ring B.
(ii) Homoannular cyclohexadiene component adds +35 nm (ring atoms must be counted separately as
substituents)
(iii) Solvent Correction: water = 8; methanol/ethanol = 0; ether = +7; hexane/cyclohexane = +11
max (calculated) = Base + Substituent Contributions and Corrections

Some examples that illustrate these rules follow.




End of this supplementary topic