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Spectrophotometric Methods
2007
By the end of this part of the course, you should be able to:
If you have difficulty to understand above topics, find extra reading materials!
Or discuss with me after the lecture.
c/n=
Therefore:
Energy is inversely proportional to wavelength
but proportional to wavenumber
Frequency Scanning Techniques: a few definitions
Emission method: source of light is sample
Absorption method: intensities of a source with and without the sample in place are compared
In quantitative analysis:
common to work at 1 wavelength
running a spectrum is an important initial step (to select best conditions)
Regions of Electromagnetic Spectrum-the colour of light
Fig. 18-2
Electronic structures of simple molecule
Energy
Excited state
Singlet
S1
T1 Excited state
Vibration states
Triplet
D
Dissociated states
S0 Bond length
Ground state
Interaction between photon and molecule
S0 S1 transition
S1
T1
S1
T1
UV-vis
D
A F
P
S0
IR
S0
Key concept from energy diagram
Electronic structures
Internal conversion
Intersystem crossing
Luminescence-fluorescence/phosphorescence
Type of optical spectroscopy
UV-vis absorption spectroscopy (UV-Vis)
FT-IR absorption/transmission spectroscopy (FTIR)
Atomic absorption spectroscopy (AAS)
Atomic fluorescence spectroscopy (AFS)
X-ray fluorescence spectroscopy (XFS)
Monochromator design
Instrument principle
Quantitative methods
Optical spectrophotometer components
Excitation sources
(a)
(c)
(b)
Fig. 13-12, pg. 315 "Instrument designs for photometers and spectrophotometers
Light sources Brightness
Line width
What is the important properties of a source? Background
Black-body radiation for vis and IR but not UV Stability
- a tungsten lamp is an excellent source of black-body radiation Lifetime
- operates at 3000 K
- produces from 320 to 2500 nm
( How much in cm-1, J, Hz and eV?)
For UV:
- a common lamp is a deuterium arc lamp
- electric discharge causes D2 to dissociate and emit UV radiation (160 325 nm)
- other good sources are:
Xe (250 1000 nm)
Hg (280 1400 nm)
Lasers:
- high power
- very good for studying reactions
- narrow line width
- coherence
- can fine-tune the desired wavelength (but choice of wavelength is limited)
- expensive
Sample a source containers:
for UV: quartz (wont block out the light)
for vis: glass [ 800nm (red) to 400 nm (violet)]
for IR: NaCl (to or 15384 nm or 650 cm-1)
KBr (to 22222 nm or 450 cm-1)
CsI (to 50000 nm or 200 cm-1)
Criteria
High transmission
Chemically inert
Mechanically strong
Monochromators
Early spectrophotometers used prisms
- quartz for UV
- glass for vis and IR Why?
http://www.mrfiber.com/images/
cddiffract.jpg
10mx10m
http://www.veeco.com/library/nanotheater_detail.php?
type=application&id=331&app_id=34
Monochromators: contd
What is the purpose of concave mirrors?
Polychromatic radiation enters
The light is collimated the first concave mirror
Reflection grating diffracts different
wavelengths at different angles
http://oco.jpl.nasa.gov/images/grating_spec-br.jpg
Interference in diffraction
d sin()+d sin()=n
d
Bragg condition
Phase relationship
>0
<0 n=1, 2, 3 In-phase
n=1/2, 3/2, 5/2 out-phase
Monochromators: reflection grating
Monochromators: reflection grating
Each wavelength is diffracted off the grating at a different angle
Groove dimensions and spacings are on the order of the wavelength in question
In order for the emerging light to be of any use, the emerging light beams must be in phase
with each other
Angular resolution:
As: d sin()+d sin()=n
So: n =d cos()
Therefore: =n/[d cos()]
What does this mean?
Monochromators: slit
Bottom line:
- it is usually possible to arrange slits and mirrors
so that the first order (n = 1) reflection is separated
- a waveband of ca. 0.2 nm is obtainable
However, the slit width determines the resolution and signal to noise ratio
Large slit width: more energy reaching the detector higher signal:noise
Small slit width: less energy reaching the detector BUT better resolution!
Detectors : Radiation-----charger converter
Choice of detector depends upon what wavelength you are studying
Want the best response for the wavelength (or wavelength range) that you are studying
In a single-beam spectrophotometer, the 100% transmittance control must be adjusted each time the
wavelength is changed
In a double-beam spectrophotometer, this is done for you!
Photomultiplier-single channel, but very high sensitivity
A diode is a pn junction:
under forward bias, current flows from n-Si to p-Si
under reverse bias, no current flows
boundary is called a depletion layer or region
Photodiode Array
- Electrons excited by light partially discharge the condenser
- The grating polychromator disperses the light into the component wavelengths
- Resolution depends upon the distance between the diodes and amount of dispersion
No moving parts!
Simple mechanical and optical design, very compact.
Photodiode Array Spectrophotometers
vs Dispersive Spectrophotometers
Photodiode Array
Spectrophotometer:
- no moving parts rugged
- faster spectral acquisition (ca.
1 sec)
- not dramatically affect by room
light
Fluorescence vs phosphorescence
Absorption vs emission
Why?
Fluorescence spectroscopy
Fluorescence spectroscopy
Light source Beam
splitter Q: why the emission is
measured at 90 relative to
Excitation
monochromator
sample the excitation?
ght
li
of Emission
8% Monochromator
Reference
diode
Emission spectrum: hold the excitation wavelength steady and measure the emission at
various wavelengths
Excitation spectrum: vary the excitation wavelength and vary the wavelength measured for
the emitted light
Fluorescence spectroscopy: well defined molecules
Summary of spectrophotometric techniques
Describe the main components of a spectrophotometer and distinguish between single
double beam instruments
Describe suitable sources for ultraviolet (UV)/visible (vis), infra red (IR) and atomic
absorption (AA) instruments
Describe and assess advantages and disadvantages of various monochromators e.g.
Prism, diffraction gratings
Explain how to asses the quality of grating
Explain how photomultipliers and diode detectors work
Explain the advantage of multiplex detecting
Describe the luminescence spectroscopy and energy transfer process
Compare the emission and absorption spectrum