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Spectrophotometry Fundamentals
Spectrophotometry Fundamentals
Key Concepts
Lamberts Law of Absorption
Beers Law
Beer-Lambert Law
Absorption Cross-Sections
Photometric quantities
Spectrophotometer
The Cary 50 Spectrophotometer
I0
I I 0 e l
I
I ( x ) I 0 e x
I(x)
d I I d x
I I 0e
dI
I
dx
I
x
e
I0
Photo: http://www-history.mcs.st-andrews.ac.uk/history/PictDisplay/Lambert.html
I I 0e
I 010
kx
k ln 10
I
x
kx
e 10
I0
k is the path length over which the intensity is attenuated to 1/10.
I
10 k x
I0
I0
I
1
10 k l
I0
2
Then two slabs of the same absorbing material will then reduce the intensity of a beam
of light to one quarter.
I0
I
1
10 k 2 l
I0
2
1
4
And three slabs will reduce the intensity of a beam of light to one eight.
l
I0
I
1
k 3l
10
I0
2
Beers Law
Beer found that Lamberts linear decay constant k for a solution of an
absorbing substance is linearly related to its concentration c by a constant,
the absorptivity , a characteristic of the absorbing substance.
Restatement: The linear decay constant k is linear in concentration c with a
constant of proportionality .
(August Beer, 1825-1863)
k c
Typical units are: k cm1; c M (moles/liter); M1cm1
A colored absorber has an absorptivity that is dependent on wavelength of the light ().
The absorptivity is the fundamental property of a substance. This is the property that
contains the observable spectroscopic information that can be linked to quantum
mechanics (also see absorption cross section.)
Photometric Quantities
In photometry we measure the intensity of light and characterize its
change by and object or substance. This change is typically expresses as
percent transmittance or absorbance.
Transmittance (T)
Frequently when your primary
interest is the light beam
I
T
I0
I
log T
A log
I0
Beer-Lambert Law
Lamberts and Beers Laws are combined to describe the attenuation of light by a
solution. It is easy to see how the two standard photometric quantities can be written in
terms of this law.
I I 010
c x
Transmittance
Absorbance
I
T
I0
I
log T
A log
I0
A cx
T 10 c x
NC
k NC ln 10
N A C ln 10 10 3
liter
3
cm
Efficiency
The absorption efficiency Q of a particle is the ratio of its absorption cross
section C to its geometric cross section Cgeo.
Absorption efficiency is dimensionless.
C
Q
C geo
Note:
All of these quantities are in general wavelength dependent.
Our discussion has not included the mechanism (cause) of absorption and scattering.
There are many different mechanisms that cause of absorption and scattering.
Instrumentation
Spectrometer: measures I vs .
Simply measures the spectrum of the light (e.g. emission spectroscopy).
The Spectrophotometer
Measures absorbance as a function of wavelength
Components: light source, monochromator, sample cell, detector, optical
system.
sample cell
slit
diffraction grating
monochromator
light source
detector
monochromator
balance the forces:
detector
sample
light
source
www.varianinc.com
I
raw I zero
T
I 0 raw I 0 zero
A log T
Applications of
Spectrophotometry
Spectroscopy
Chemical Analysis: