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scopy
• Spectroscopy: study EMV and matter
• Spectroscopy method for determination of organic molec
ular structure
• Mass spectroscopy
• Ultraviolet-visible spectroscopy
• Infrared spectroscopy
• Nuclear magnetic resonance spectroscopy
1
Interaction of electromagnetic radia
tion energy and matter
• When EMR is directed at a substance, the radiatio
n can be:
– Absorbed
– Transmitted
– Reflected
2
Interaction of electromagnetic radia
tion energy and matter
• When EMR is directed at a substance, the radiatio
n can be:
– Absorbed
– Transmitted
– Reflected
3
Electromagnetic Radiation
4
Mathematical Relationships
c = E = h E = hc /
6
Interaction of electromagnetic radia
tion energy and matter
• Molecules exist only in discrete states that
correspond to discrete energy content.
• The EMR energy that is absorbed is quanti
zed and brings about certain specific chan
ges in the molecule.
– electronic transitions (UV-vis)
– vibrations (IR)
– rotations (IR)
7
Interaction of electromagnetic radia
tion energy and matter
• Exact energies absorbed by a molecule ar
e highly characteristic of the structure and
are unique for each compound.
– spectroscopic “fingerprint”
12
Electronic molecular orbital energies
s s* transition in vacuum UV (<180 nm) not use in organic C-C 135 nm
n s* saturated compounds with non-bonding electrons (N, O, S, hal)
~ 150-250 nm
e ~ 100-3000 ( not strong)
n p*, p p* requires unsaturated functional groups (eq. double bonds)
most commonly used, energy good range for UV/Vis
~ 200 - 700 nm
n p* : e ~ 10-100
p p*: e ~ 1000 – 10,000
RELATION OF STRUCTURE AND
e
Name Structure max e
but-1-en-3-yne 219 7,600
vacuum UV
most compounds of atmosphere absorb in this range, so
difficult to work with.
usually concerned with functional groups with relatively low
excitation energies (190 850 nm).
i) s, p, n electrons
Example: Formaldehyde
UV-visible Spectroscopy
• The ground state of an organic molecule can contain valence ele
ctrons in three principal types of molecular orbitals:
s (sigma)
C:H
p (pi) C::C
n (non-bonding)
18
Difference transition of n to pi* and pi to pi*
The effect of transition to λ
Absorption Characteristics of Some Common Chromophores
Chromophore Example Solvent max (nm) emax Type of tra
nsition
Alkene C
H
61
3H
CC
H2 n-Heptane 177 13,000 pp*
Alkyne n-Heptane 178 10,000 pp*
C
H
5C
1
1 CC
H
3 196 2,000 _
225 160
_
Carbonyl O n-Hexane 186 1,000 ns*
280 16 np*
CH3CCH3
O n-Hexane 180 Large
293 12
ns*
CH 3 CH np*
Carboxyl O Ethanol 204 41 np*
CH3COH
Amido O Water 214 60 np*
CH3CNH2
Azo H
CNN
CH Ethanol 339 5 np*
3 3
Nitro CH3NO2 Isooctane 280 22 np*
Nitroso C4H9NO Ethyl ether 300 100 _
665 20 np*
Nitrate C2H5ONO2 Dioxane 270 12 np*
Other Examples of Some Com
mon Chromophores
UV-visible Spectroscopy
• Electrons in sigma bonds (single bonds) are too tightly bound
to be promoted to a higher energy level by UV-visible radiatio
n.
– alkanes, alcohols, alkyl halides, simple alkenes do not absorb in the UV
23
1). For Compounds with Multiple Chromophores:
If greater then one single bond apart
- e are additive
- constant
If conjugated
- shifts to higher ’s (red shift)
(c) Aqueous
3) Solvent can also absorb in UV-vis spectrum.
3) Can obtain some functional group information for certain types of compounds..
- weak band at 280-290 nm that is shifted to shorter ’s with an increase
in polarity (solvent) implies a carbonyl group.
acetone:
in hexane, max = 279 nm (e = 15)
in water, max = 264.5 nm
- solvent effects due to stabilization or destabilization of ground or excited
states, changing the energy gap.
since most transitions result in an excited state that is more
polar than the ground state
- 260 nm with some fine structure implies an aromatic ring.
Benzene in heptane
More complex ring systems shift to higher
’s (red shift) similar to conjugated alkenes
The effect of pH to λ
Borax buffer: 0.05M (pH 9.2 ~ 239 nm) and A 452
NaOH : pH 13 ~ 254 and A = 342
The effect of conjugation bond to absorption
PIGMENTS ON VISIBLE LIGHT
UV Spectral Nomenclature
Λ max SOME ORGANIC COMPOUNDS
The Applications of UV for determining of Λ max
of organic compounds
EMPIRICAL RULES FOR ABSORPTION WAVELENGTHS
OF CONJUGATED SYSTEMS
SOME EXAMPLES THAT ILLUSTRATE THESE RULES FOLLOW
SOME EXAMPLES TO PREDICT THE Λ max
SOME WORKED EXAMPLES TO PREDICT THE Λ max
2. Woodward-Fieser rules for calculating π to π* of conjugated
Carbonyl compounds
Similar for enones
Similar for acid and ester
C) Quantitative Analysis (Beer’s Law):
1) Widely used for Quantitative Analysis Characterization
- wide range of applications (organic & inorganic)
- limit of detection 10-4 to 10-5 M (10-6 to 10-7M; current)
- moderate to high selectivity
- typical accuracy of 1-3% ( can be ~0.1%)
- easy to perform, cheap
2) Strategies
a) absorbing species
- detect both organic and inorganic compounds containing any of
these species (all the previous examples)
b) non- absorbing species
- react with reagent that forms colored product
- can also use for absorbing species to lower limit of detection
- items to consider:
, pH, temperature, ionic strength
- prepare standard curve (match standards and samples as much as
possible)
Complex
(red)
- used for analytes in a complex matrix where interferences in the UV/Vis for the
analyte will occur: i.e. blood, sediment, human serum, etc..
- Method:
(1) Prepare several identical aliquots, Vx, of the unknown sample.
Note: This method assumes a linear relationship between instrument response and sample concen
tration.
(3) Dilute each solution to an equal volume, Vt.
m = y/ x
Instrument Response ( S )
kV
scs kVc
b = y-intercept S x
x
V V
S s
mVb t t
(V s ) 0
SV
1 c
x
C s s
Vs (S2S)
1V x
Where:
S = signal or instrument response
k = proportionality constant
Vs = volume of standard added
bcs cs = concentration of the standard
cx Vx = volume of the sample aliquot
mVx cx = concentration of the sample
Vt = total volume of diluted solutions
Note: assumes a linear relationship between instrument response and sample concentration.
c) Analysis of Mixtures
- use two different ’s with different e’s
Calculate the concentrations of A and B in solutions that yielded an absorbance of 0.439 at 475 nm a
nd 1.025 at 700 nm in a 2.50-cm cell.