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Analysis
UV-Vis spectroscopy
Electronic spectroscopy
UV_VIS
Principle
Electromagnetic
radiation
Sample
Quanlitative
Quantitative
(Absorption or emission)
UV_VIS
Electromagnetic radiation
Wave Properties
Particle Properties
Phng truyen
Direction of propagation
UV_VIS
E = h = hc/ = h.c.
E (eV, kcal/mol)
h: Planck constant = 6,626.10 34
J.s = 6,626.10 27 erg.s = 6,59 eV.s
Electromagnetic spectrum
UV-VIS
Electromagnetic spectrum
IR
UV-VIS
(Gamma rays)
Violet
X rays
Blue
380 nm
E = h = hc/ = hc
UV_VIS
Visible
Ultraviolet
Green
Yellow
Colorimetry
Infrared
Orange
NMR
Radio waves
Red
760 nm
Visible lights
UV-VIS
UV_VIS
i cng quang
Principle
Electromagnetic
radiation
Sample
Quanlitative
Quantitative
(Absorption or emission)
UV_VIS
E1
Ee = E1 E0
E0
E = Ee + Ev + Er
Ee : (electron energy)
Ev: (vibration energy)
Er: (rotation energy)
UV-Vis spectroscopy
Electronic spectroscopy
UV_VIS
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Instrument
UV_VIS
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Instrument
UV_VIS
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Instrument
UV_VIS
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Intrument
UV_VIS
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UV/VIS instrument
i cng quang
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Instrument
UV_VIS
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Instrument
UV_VIS
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Instrument
UV_VIS
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Grating
monochromator
polychromatic
radiation
monochromatic
radiation
UV_VIS
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Source
UV_VIS
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Detectors
UV_VIS
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VIS spectrum
UV_VIS
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IR spectrum
UV_VIS
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UV_VIS
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* Unoccupied level
Atommic Orbital
Occupied level
Molecular orbitals
* Energy
*
n - *
- *
-*
n-*
UV_VIS
Molecular orbitals
Chromophores
Chromophores are groups of atoms within a molecule,
which absorb electromagnetic radiation.
The most important chromophores are:
Conjugated double bonds, such as:
O
UV_VIS
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Notation of transition
max(nm)
~150
n *
n *
~185
~195
n * ~195
~300
~190
-bonded electrons
lone pair electrons
-O-N<
-S>C=O
>C=O
n *
n *
-bonded electrons
>C=C< (isolated)
*
>C=O
*
UV_VIS
of these,
mainly the
>C=O
absorption can
be seen in a
normal UV
spectrum
~190
~190
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*
A
B
2
isolated
double bond
UV_VIS
1
two conjugated
double bonds
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UV_VIS
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UV_VIS
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Absorption
Chromophore
Auxochrome
Bathochromic shift, red shift hypsochromic effect, blue
shift
Hyperchromic effect hypochromic effect
UV_VIS
31
Solvent Choice
The choice of solvent in UV-Vis detection is
dependent on a few factors:
The solvent should not absorb light in the same
wavelength region as the substance that is being
analysed.
The solvent should be transparent at the wavelengths
that are being used in the analysis.
The solvent should not form a complex with the
analyte, subsequently disturbing the absorption
spectrum.
The solvent can be used to shift the absorption
wavelengths to either longer or shorter transition
wavelengths.
UV_VIS
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Solvent Choice
Solvent
Water
Ethanol
Hexane
Cyclohexane
Methanol
Diethyl ether
Acetonitrile
Tetrahydrofuran
Dichloromethane
Chloroform
Carbon tetrachloride
Benzene
UV_VIS
205
210
210
210
210
210
210
235
245
265
280
solvents of choice
- no significant
interference
220
33
Effect of Solvent
The solvent can influence the position (max) and
the molar absorptivity (max) of the absorbance
spectra, through changes in:
pH
Polarity
Electrolyte concentration
UV_VIS
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In Conclusion
Alkanes, alcohols and ethers cannot be observed
in UV-Vis, as the transitions involved are *
and n*
Ketones generally show weak n* transitions
and are visible in the UV region
Dienes and enons show strong * absorptions
and are also visible in the UV region.
UV_VIS
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IO = IR + I A + I T = IA + I T
I
T (transmittance)
T = IT/Io or
T% = 100 x IT/Io
UV_VIS
A (Absorbance)
A = log I0/IT = log 1/T = log
100/ T% = 2 log T%
36
A=bC
Io
IA
IT
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Example
A 5.00 104 M solution of an analyte is placed in a sample
cell that has a pathlength of 1.00 cm. When measured at a
wavelength of 490 nm, the absorbance of the solution is
found to be 0.338. What is the analytes molar
absorptivity at this wavelength?
UV_VIS
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Limitations to
Beers Law
Concentration
pH or dilution
Solvent
Temperature
Time
Ligand
UV_VIS
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Applications
Qualitative
Quantitative
One component
Multiple component
UV_VIS
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VIS spectrum
UV-VIS
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UV-VIS
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UV-VIS
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C0
C1
C2
C3
A0
A1
A2
A3
C4
A4
C5
A5
M0
M1
A(M0)
A(M1)
A5
A4
A3
A2
Cm
A1
UV_VIS
C1
C2 C3 C4
C5
C, mol/L
44
ppm Fe
0.00
1.00
2.00
3.00
4.00
unknown
UV_VIS
Absorbance
0.000
0.183
0.364
0.546
0.727
0.269
45
UV_VIS
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A1 A1 A1 1 bC 1 bC
I
II
II
II
A2 II1 A1 II2
I II I II
2
A2 1 A1 2
II
C
II
II
I II I II
2
UV_VIS
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UV_VIS
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UV_VIS
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