Tutorial on Atomic Absorption

Spectroscopy and Beer Lambert Law

Prepared by
Lawrence Kok
http://lawrencekok.blogspot.
com

Continuous Spectrum Vs Line Spectrum

Continuous
Continuous Spectrum
Spectrum ::
Light
Light spectrum
spectrum with
with all
all
wavelength/frequency
wavelength/frequency
Emission
Emission Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/
wavelength/ frequency
frequency
Emitted
Emitted when
when excited
excited electrons
electrons drop
drop from
from higher
higher to
to lower
lower
energy
level
energy level
Absorption
Absorption Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/frequency
wavelength/frequency
Absorbed
Absorbed when
when ground
ground state
state electrons
electrons are
are excited
excited

http://www.astrophys-assist.com/educate/orion/orion02.htm

Continuous Spectrum Vs Line Spectrum

Continuous
Continuous Spectrum
Spectrum ::
Light
Light spectrum
spectrum with
with all
all
wavelength/frequency
wavelength/frequency
Emission
Emission Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/
wavelength/ frequency
frequency
Emitted
Emitted when
when excited
excited electrons
electrons drop
drop from
from higher
higher to
to lower
lower
energy
level
energy level
Absorption
Absorption Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/frequency
wavelength/frequency
Absorbed
Absorbed when
when ground
ground state
state electrons
electrons are
are excited
excited

Atomic Emission Spectroscopy

Electrons from excited state

Emit radiation
when drop to ground
state
Radiation emitted

Emission Spectrum

http://www.astrophys-assist.com/educate/orion/orion02.htm

Continuous Spectrum Vs Line Spectrum

Continuous
Continuous Spectrum
Spectrum ::
Light
Light spectrum
spectrum with
with all
all
wavelength/frequency
wavelength/frequency
Emission
Emission Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/
wavelength/ frequency
frequency
Emitted
Emitted when
when excited
excited electrons
electrons drop
drop from
from higher
higher to
to lower
lower
energy
level
energy level
Absorption
Absorption Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/frequency
wavelength/frequency
Absorbed
Absorbed when
when ground
ground state
state electrons
electrons are
are excited
excited

Atomic Emission Spectroscopy Vs Atomic Absorption Spectroscopy

Electrons from excited state

Emit radiation
when drop to ground
state

Electrons in excited state

Radiation absorbed
Radiation emitted

Emission Spectrum

Absorb radiation
to excited state

Electrons from ground state

http://www.astrophys-assist.com/educate/orion/orion02.htm

Continuous Spectrum Vs Line Spectrum

Continuous
Continuous Spectrum
Spectrum ::
Light
Light spectrum
spectrum with
with all
all
wavelength/frequency
wavelength/frequency
Emission
Emission Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/
wavelength/ frequency
frequency
Emitted
Emitted when
when excited
excited electrons
electrons drop
drop from
from higher
higher to
to lower
lower
energy
level
energy level
Absorption
Absorption Line
Line Spectrum
Spectrum ::
Spectrum
Spectrum with
with discrete
discrete wavelength/frequency
wavelength/frequency
Absorbed
Absorbed when
when ground
ground state
state electrons
electrons are
are excited
excited

Atomic Emission Spectroscopy Vs Atomic Absorption Spectroscopy

Electrons from excited state

Excited state

Electrons in excited state

Emit radiation
when drop to ground
state

Radiation absorbed
Radiation emitted
Absorb radiation
to excited state

Emission Spectrum

Ground state

http://www.astrophys-assist.com/educate/orion/orion02.htm

Electrons from ground state

Atomic Emission Spectroscopy

Line Emission Spectra for Hydrogen

Energy
Energy supplied
supplied to
to atoms
atoms
Electrons
are
excited
from
Electrons are excited from ground
ground to
to excited
excited states
states

UV
UV
region
region
Lyman
Lyman
Series
Series
n=
n=
n=
n=
1
1
Emission

Visible
Visible
region
region
Balmer
Balmer
Series
Series
n=
n=
n=
n=
2
2
Spectra

Electrons
Electrons exist
exist in
in fixed
fixed energy
energy level
level (quantum)
(quantum)
Electrons
drop
from
higher
to
lower,
Electrons drop from higher to lower,
emitting
emitting energy
energy of
of particular
particular wavelength/frequency
wavelength/frequency
Higher
Higher the
the energy
energy level,
level, smaller
smaller the
the difference
difference in
in energy
energy
between
successive
energy
level.
between successive energy level.
Spectrum
Spectrum will
will converge(get
converge(get closer)
closer) with
with increasing
increasing frequency
frequency
Lines
in
spectrum
convergeenergy
levels
Lines in spectrum converge- energy levels also
also converge
converge
Ionisation
Ionisation energy
energy can
can be
be determined
determined (Limit
(Limit of
of convergence)
convergence)

IR
IR region
region
Paschen
Paschen
Series
Series
n=
n=
n=
n= 3
3

omic
omic Emission Spectra
Energy
Energy supplied
supplied
Electrons
Electrons surround
surround nucleus
nucleus in
in allowed
allowed energy
energy states
states (quantum)
(quantum)
Excited
Excited electron
electron returning
returning to
to lower
lower energy
energy level,
level, photon
photon of
of light
light with
with
iscrete
energy/wavelength(colour)
will
be
given
out.
iscrete energy/wavelength(colour) will be given out.
Light
Light pass
pass through
through a
a spectroscope,
spectroscope, with
with a
a prism
prism to
to separate
separate
ut
different
colours
ut different colours
Line
Line emission
emission spectra
spectra is
is produced.
produced.

Line emission spectra for different elements can be found here

N= 6-2
410nm

N= 5-2
434nm

N= 4-2
486nm

N = 3-2,
656nm

Visible region- Balmer Series

Atomic Emission Spectroscopy

nciple
nciple of
of Atomic
Atomic Absorption
Absorption Spectroscopy:
Spectroscopy:
AAS
spectroscopy
uses
absorption
AAS spectroscopy uses absorption of
of light
light to
to measure
measure the
the concentration
concentration of
of gas-phase
gas-phase atoms.
atoms.
Sample
Sample in
in liquids/solids
liquids/solids must
must be
be vaporized
vaporized in
in a
a flame/graphite
flame/graphite furnace
furnace to
to gaseous
gaseous atomic
atomic vapour.
vapour.
Atoms
absorb
UV
/
visible
light
causing
electronic
transitions
to
higher
energy
levels.
Atoms absorb UV / visible light causing electronic transitions to higher energy levels.
Analyte
Analyte concentration
concentration determined
determined from
from the
the amount
amount of
of absorption.
absorption.
Amount
of
light
absorbed
=
(Ratio
of
light
intensity
transmitted/incident
Amount of light absorbed = (Ratio of light intensity transmitted/incident light)
light)
Amount
of
light
absorbed
=
directly
proportional
to
concentration
of
analytes/ions/atoms.
Amount of light absorbed = directly proportional to concentration of analytes/ions/atoms.

Electrons in excited state

Radiation absorbed

Absorb radiation
to excited state

Electrons from ground state

Uses of Atomic Absorption Spectroscopy (AAS)

1)
1)
2)
2)
)
)
)
)
)
)
)
)
)
)

Detection
Detection of
of metal
metal ions
ions in
in small
small quantities
quantities (ppm/ppb)
(ppm/ppb)
Detection
of
Al,
Ca,
Cr,
Fe
in
various
sample
Detection of Al, Ca, Cr, Fe in various sample
Al
Al -- Blood
Blood serum
serum
Ca

Blood
Ca Blood serum
serum
Cu
Cu Copper
Copper based
based alloys
alloys
Cr
Cr Sea
Sea water
water
Fe
Fe -- Plants
Plants
http://csep10.phys.utk.edu/astr162/lect/light/absorption.html

Parts of Atomic Absorption Spectroscopy

Hollow Cathode Lamp

Emits light of specific wavelength
to be absorbed by element

Monochromator
Select light with specific frequency/wavelength of interest
Provide the specific light which will be absorb by metal

Atomiser
Atomise sample to atomic
state
Atom vapour (gas phase)
produced

Fuel
Burning mixture of ethyne with O2
(combustion mix)
Sample dehydrated/solvent removed
http://health-hazard-mercury.blogspot.kr/

Detector

Measure the decrease in intensity of light absorb by analyte

Convert the decrease in light intensity into electrical signal using
photomultiplier

How AAS works to measure the Copper concentration in a sample ?

1
1

Hollow Cathode Copper Lamp

Emits light of specific wavelength
to be absorbed by copper element

http://health-hazard-mercury.blogspot.kr/

How AAS works to measure the Copper concentration in a sample ?

1
1

Hollow Cathode Copper Lamp

Emits light of specific wavelength
to be absorbed by copper element

Hollow cathode lamp (Cu)

Made of element (Cu) and use to determine Cu conc in sample
Light source emits light of specific wavelength/frequency which will be absorb by
the sample/element

Light source of 325nm used, Cu atoms in sample will absorb that particular
wavelength to be excited.

Light , = 325nm emitted

Atomic Cu vapour absorb light, = 325nm

http://health-hazard-mercury.blogspot.kr/

How AAS works to measure the Copper concentration in a sample ?

2
2
/ Atomizer

uel Air/acetylene flame, Temp of 2300C

esolvation (drying) Solvent evaporated and dry vapour sample produced
tomization Sample molecules dissociate into free atoms /ions and converted finally to atomic vapour
opper atoms vapour state produced .

Light , = 325nm emitted

Atomic Cu vapour absorb light, = 325nm

http://health-hazard-mercury.blogspot.kr/

How AAS works to measure the Copper concentration in a sample ?

3
3

Detector

Atoms absorb UV/visible light, exciting its electrons (electronic transition) to

higher energy levels.
Measure the decrease in light intensity (emitted by Cu lamp) after being
absorbed by copper atoms
Amount light absorbed is directly proportional to the concentration of the atoms.

http://health-hazard-mercury.blogspot.kr/

How AAS works to measure the Copper concentration in a sample ?

3
3

Detector

Atoms absorb UV/visible light, exciting its electrons (electronic transition) to

higher energy levels.
Measure the decrease in light intensity (emitted by Cu lamp) after being
absorbed by copper atoms
Amount light absorbed is directly proportional to the concentration of the atoms.

Initial
Light
Intensity,
Io

Light , = 325nm emitted

Atomic Cu vapour absorb light, = 325nm

http://health-hazard-mercury.blogspot.kr/

Transmitted
Light Intensity

Decease in Light Intensity due to Absorption

Beers- Lambert Law and Standard Calibration Curve

Beers Lambert Law
Apply for diluted solution

Absorbance, A Concentration
Absorbance, A = log10 (Io/I) = lc
Transmittance, T = Fraction of incident radiation (I) transmitted by the solution, T= I/I o

Beers- Lambert Law and Standard Calibration Curve

Beers Lambert Law
Apply for diluted solution

Absorbance, A Concentration
Absorbance, A = log10 (Io/I) = lc
Transmittance, T = Fraction of incident radiation (I) transmitted by the solution, T= I/I o

How Absorbance equation is derived ?

Abs =
lc

Beers- Lambert Law and Standard Calibration Curve

Beers Lambert Law
Apply for diluted solution

Absorbance, A Concentration
Absorbance, A = log10 (Io/I) = lc
Transmittance, T = Fraction of incident radiation (I) transmitted by the solution, T= I/I o

How Absorbance equation is derived ?

Abs =
lc
Amount light absorb depend
on

= Molar extinction of
compound
c= Concentration
l = path length of absorbing
solution

If and l = constant

Abs Conc analyte

Abs = Conc

Abs
Conc, c

Molar extinction of compound, :

Measure the strength of absorption of

compound

Higher = Higher Absorbance

Compound with high = effective at absorbing

light even when low conc is used.
Path Length, l:

Longer path length Higher Absorbance

Concentration, c:

Higher conc of analyte Higher Absorbance

Determine the unknown concentration of Pb2+ using Beer-Lambert Law

1
1
Preparing
a Standard Calibration Curve for Lead ions concentration

Determine the unknown concentration of Pb2+ using Beer-Lambert Law

1
1
Preparing
a Standard Calibration Curve for Lead ions concentration

Plot
2
2 a Standard calibration curve of Abs vs Conc
Abs

Conc

Determine the unknown concentration of Pb2+ using Beer-Lambert Law

1
1
Preparing
a Standard Calibration Curve for Lead ions concentration

Plot
2
2 a Standard calibration curve of Abs vs Conc
Abs

Conc

3
3 Measure Absorbance of unknown
(Pb2+) sample
Abs

Abs = 0.340

Conc = 0.310
Conc

Determine the unknown concentration of Pb2+ using Beer-Lambert Law

1
1
Preparing
a Standard Calibration Curve for Lead ions concentration

Plot
2
2 a Standard calibration curve of Abs vs Conc
Abs

Conc

3
3 Measure Absorbance of unknown
(Pb2+) sample

Determine
4
Conc of unknown (Pb2+) by interpolation
4

Abs

Abs = 0.340

Conc = 0.310
Conc

Amount of light absorb Conc of Pb vapour in flame

Absorbance Conc of Pb atoms
Absorbance, A = lc
= molar absorptivity
l = path length constant
c = conc of sample
Since and l are constant
A = c ( Absorbance directly proportional to Conc)

Determine the concentration of unknown using Beer-Lambert Law

Question, Q1

7.25 x 10-5M solution of X has a transmittance of 44.1% when measured in a 2.10cm cell at
wavelength of 525nm.
Calculate the
a) Absorbance, A
b) Molar absorptivity,

Question, Q2

100dm3 of contaminated water was reduced by boiling to 7.50dm3. The reduced volume was
tested, it had absorbance of 0.55. Calculate the conc of Pb 2+ ions (mgdm3) in original sample.

Determine the concentration of unknown using Beer-Lambert Law

Question, Q1

7.25 x 10-5M solution of X has a transmittance of 44.1% when measured in a 2.10cm cell at
wavelength of 525nm.
Calculate the
a) Absorbance, A
b) Molar absorptivity,
Answer to Q1

a) Absorbance = -log10 T or Absorbance = lc

b) Molar absorptivity,

Abs = - log10 T
= A/bc =
0.3554 / ((2.10 x 7.25 x 10-5)
= -Q2log10 0.441
= 2.33 x
Question,
-1
103 L mol
cm-1
3
3
100dm
of
contaminated
. The reduced volume was
= -(-0.3554)
= 0.355water was reduced by boiling to 7.50dm
2+
tested, it had absorbance of 0.55. Calculate the conc of Pb ions (mgdm3) in original sample.
Answer to Q2

Amount /moles before heating = Amount/moles after heating

Moles before = Moles after
MxV=MxV
M x 100 = 1.15 x 7.50
M = (1.15 x 7.50)/100
M = 8.63 x 10-2 mgdm-3

V= 100dm3
M= ?

V= 7.50dm3
M= 1.15mg/dm3

Conc of Pb2+ from graph 1.15mgdm-3

Acknowledgements
Thanks to source of pictures and video used
in this presentation
Thanks to Creative Commons for excellent
contribution on licenses
http://creativecommons.org/licenses/

Prepared by Lawrence Kok

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