Académique Documents
Professionnel Documents
Culture Documents
FT-NMR
X-Ray Spectroscopy X-ray Crystallography
Radio waves
X-rays X-rays
Atomic Absorption
FT-IR Raman FT-NMR
UV-vis region
IR/Microwave IR/UV Radio waves
X-Ray Spectroscopy
X-ray Crystallography
X-rays
X-rays
Elemental Analysis
3-D structure Anaylysis
Different Spectroscopies
UV-vis electronic states of valence e/dorbital transitions for solvated transition metals Fluorescence emission of UV/vis by certain molecules FT-IR vibrational transitions of molecules FT-NMR nuclear spin transitions X-Ray Spectroscopy electronic transitions of core electrons
Quantitative Spectroscopy
Beers Law Al1 = el1bc e is molar absorptivity (unique for a given compound at l1) b is path length c concentration
Beers Law
slit source detector cuvette
A = -logT = log(P0/P) = ebc T = Psolution/Psolvent = P/P0 Works for monochromatic light Compound x has a unique e at different wavelengths
Standard Practice
Prepare standards of known concentration Measure absorbance at lmax Plot A vs. concentration Obtain slope Use slope (and intercept) to determine the concentration of the analyte in the unknown
y = 0.02x
20.0
40.0
60.0
concentration (uM)
UV-Vis Spectroscopy
UV- organic molecules
Outer electron bonding transitions conjugation
Instrumentation
Molecules have quantized energy levels: ex. electronic energy levels. hv energy energy
} = hv
Q: Where do these quantized energy levels come from? A: The electronic configurations associated with bonding.
Each electronic energy level (configuration) has associated with it the many vibrational energy levels we examined with IR.
Broad spectra
Overlapping vibrational and rotational peaks Solvent effects
s* p*
2p
p s
2p
s* 2s
2s s
Ethane
C C
s*
hv
s* s
s
H C C H H H H H
s*
Absorptions having lmax < 200 nm are difficult to observe because everything (including quartz glass and air) absorbs in this spectral region.
C C
s* p*
hv
s* p*
= hv =hc/l
p s
p s
p*
C O
s* p*
n
hv
s* p*
n
p s
n
p s
p*
The n to pi* transition is at even lower wavelengths but is not as strong as pi to pi* transitions. It is said to be forbidden. Example: Acetone: ns* lmax = 188 nm ; = 1860 np* lmax = 279 nm ; = 15
C C
ss*
pp*
H
135 nm
165 nm 183 nm 150 nm 188 nm 279 nm
180 nm
C C
C O
C O
weak
weak
C O
A
279 nm
Conjugated systems:
C C
LUMO
HOMO
Preferred transition is between Highest Occupied Molecular Orbital (HOMO) and Lowest Unoccupied Molecular Orbital (LUMO).
Note: Additional conjugation (double bonds) lowers the HOMOLUMO energy gap: Example: 1,3 butadiene: lmax = 217 nm ; = 21,000 1,3,5-hexatriene lmax = 258 nm ; = 35,000
Lycopene:
lmax(Actual) = 474.
Octahedral Geometry
H2O H2O
H2O
Co2+
H2O H2O
H2O
Instrumentation
Fixed wavelength instruments Scanning instruments Diode Array Instruments
Scanning Instrument
Scanning Instrument monochromator
slit
slit
sources
Tungten lamp (350-2500 nm) Deuterium (200-400 nm) Xenon Arc lamps (200-1000 nm)
Monochromator
Braggs law, nl = d(sin i + sin r) Angular dispersion, dr/dl = n / d(cos r)
Resolution, R = l/l=nN, resolution is extended by concave mirrors to refocus the divergent beam at the exit slit
Sample holder
Visible; can be plastic or glass UV; you must use quartz
slit
monochromator
Advantages/disadvantages
Scanning instrument
High spectral resolution (63000), l/l Long data acquisition time (several minutes) Low throughput
Diode array
Fast acquisition time (a couple of seconds), compatible with on-line separations High throughput (no slits) Low resolution (2 nm)
HPLC-UV
HPLC Pump
Mobile phase
Sample loop
6-port valve