Atomic Absorption

Spectroscopy

Prof Drs Mudasir, M.Eng.,

Ph.D
Chemistry Department, UGM
1

Atomic Absorption
Spectroscopy
AAS is commonly used for metal analysis
A solution of a metal compound is
sprayed into a flame and vaporises
The metal atoms absorb light of a
specific frequency, and the amount of
light absorbed is a direct measure of the
number of atoms of the metal in the
solution
2

Atomic Absorption Spectroscopy:

Developed by Alan Walsh (below) of the
CSIRO in early 1950s.

Atomic Absorption
Spectroscopy
Uses absorption of light to measure the
concentration of gas-phase atoms.
Since samples are usually liquids or solids,
the analyte atoms must be vapourised in a
flame (or graphite furnace).

Absorption and Emission

Excited States

Ground State
Absorption Emission

Multiple
Transitions
5

Atomic Absorption
Spectroscopy
The analyte concentration is
determined from the amount of
absorption.

Atomic Absorption
Spectroscopy
The analyte concentration is determined
from the amount of absorption.

Atomic Absorption
Spectroscopy
It is possible to measure the
concentration of an absorbing species in
a sample by applying the Beer-Lambert
Law:

I
Abs log
Io

= extinction coefficient

Abs cb

Atomic Absorption
Spectroscopy
But what if is unknown?
Concentration measurements can be
made from a working curve after
calibrating the instrument with
standards of known concentration.

AAS - Calibration Curve

The instrument is calibrated before use by
testing the absorbance with solutions of known
concentration.
Consider that you wanted to test the sodium
content of bottled water.
The following data was collected using solutions
of sodium chloride of known concentration
Concentration
(ppm)

Absorbance

0.1

0.3

0.5

0.710

Calibration Curve for

Sodium
A
b
s
o
r
b
a
n
c
e

1.0

0.8

0.6

0.4

0.2

Concentration (ppm)

11

Use of Calibration curve to determine

sodium concentration {sample
absorbance = 0.65}
A
b
s
o
r
b
a
n
c
e

1.0

0.8

0.6

0.4

Concentration
Na+ = 7.3ppm

0.2

Concentration (ppm)

12

Hollow-Cathode Lamps

The electric discharge ionises rare gas

(Ne or Ar usually) atoms, which in turn,
are accelerated into the cathode and
sputter metal atoms into the gas phase.
13

Hollow-Cathode Lamps

14

Hollow-Cathode Spectrum

Harris Fig.
21-3:
Steel hollowcathode

15

Atomisation
Atomic Absorption Spectroscopy (AAS)
requires that the analyte atoms be in the
gas phase.
Vapourisation is usually performed by:
Flames
Furnaces
Plasmas
16

Flame Atomisation

Harris
Fig 214(a)

17

Flame Atomisation
Degree of atomisation is temperature
dependent.
Vary flame temperature by fuel/oxidant
mixture.
Fuel
Acetylene
Acetylene
Acetylene
Hydrogen
Hydrogen
Cyanogen

Oxidant
Temperature (K)
Air
2,400 - 2,700
Nitrous Oxide
2,900 - 3,100
Oxygen
3,300 - 3,400
Air
2,300 - 2,400
Oxygen
2,800 - 3,000
Oxygen
4,800

18

Furnaces

19

Furnaces

20

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Cold Vapor and Hydride generation Atomic Absorption

FAAS and GFAAS can measure most elements

Cannot measure: mercury, selenium and arsenic

All too volatile to be measured by flame or furnace techniques

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Cold Vapor and Hydride generation
Atomic Absorption
Cold Vapor Atomic Absorption
(CVAA) Spectroscopy for Hg
Free mercury atoms exist at room
temperature, no requirement for
heating
Sample may contain Hg0, Hg22+ or
Hg2+
In CVAA Hg is chemically reduced
to the atomic state by reaction with
a strong reducing agent (e.g.
SnCl2 or NaBH4) in a reaction flask
Hg is then carried via gas stream
to the absorption cell

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Cold Vapor and Hydride generation Atomic Absorption
Hydride Generation Atomic Absorption (HGAA) Spectroscopy for As and Se
AsH3 and SeH3 generated by reaction samples containing As and Se with NaBH4
Uses same setup as FAAS except it switches nebulizer for the hydride generation
module

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Cold Vapor and Hydride generation Atomic Absorption
Hydride Generation Atomic Absorption (HGAA) Spectroscopy for As and Se
Sample is reacted in the external hydride generator with reducing agent (NaBH4)
Hydride generated is then carried via inert gas to the sample cell in the light path of
the FAAS
Unlike CVAA product is not free atoms but AsH3 / SeH3 which are not measurable
Sample cell must be heated to dissociate the hydride into free atoms (As0) and Se0)
Higher sampling efficiency leads to
lower detection limits:
ppb vs ppm for regular FAAS and
GFAAS

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Inductively Coupled Plasma Atomic Emission (ICP-OES)

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Inductively Coupled Plasma Atomic Emission (ICP-OES)

Sample is nebulized and entrained in the flow of plasma support gas (Ar)

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Inductively Coupled Plasma Atomic Emission (ICP-OES)

Plasma torch inner tube contains the sample aerosol and Ar support gas
Radio frequency generator produces a magnetic field which sets up
an oscillating current in the ions and electrons of the support gas (Ar)
Produces high temperatures (up to 10,000 K)

Atomic Spectroscopy for Metal Analysis

Instruments for Atomic Spectroscopy
Inductively Coupled Plasma Atomic Emission (ICP-OES)

Atomizes the sample and promotes atomic and ionic transitions which are observable
at UV and visible wavelengths
Excited atoms and ions emit their characteristic radiation, which are collected by a
device that sorts the radiation by wavelength
Intensity of the emission is detected and turned into a signal that is output as
concentration

Atomic Spectroscopy for Metal Analysis

Selection of the Proper Atomic Spectroscopic Techniques
Comparison of Detection Limits and Working Range

Low detection limit is essential for trace analysis

Without low level capability sample pre-concentration is required
FAAS > ICP-OES > HGAAS > GFAAS > ICP-MS

Atomic Spectroscopy for Metal Analysis

Selection of the Proper Atomic Spectroscopic Techniques
Comparison of Detection Limits and Working Range

Atomic Spectroscopy for Metal Analysis

Selection of the Proper Atomic Spectroscopic Techniques
Comparison of Interferences and Other Considerations
Interference
Spectral: in spectroscopy, interference occurs when another emission line (e.g. from
other elements in the sample) is close to the emitted line of the test element and is
not resolved by the monochromator

Chemical: formation of undesired species during atomization

Physical: variation of instrument parameters such as uptake in the burner and

atomization efficiency (gas flow rate, sample viscosity etc.)

Atomic Spectroscopy for Metal Analysis

Selection of the Proper Atomic Spectroscopic Techniques
Comparison of Interferences and Other Considerations
Other Considerations
Cost:
FAAS < GFAAS > ICP-OES << ICP-MS

Atomic Spectroscopy for Metal Analysis

Practical Tips to Sampling
Erroneous Data and Methods of Compensation

Erroneous results arise due to one or more of the sources of interference described
above
Compensation methods: background correction, higher temperatures, release agent,
alternative wavelength, internal standard, matrix spike, etc.
e.g. Chemical interferences
Refractory salts e.g. PO43-, SO42- and silicate ion
e.g. Ca2+ forms refractory insoluble Ca3(PO4)2

Add release agent (10% lanthanum solution or EDTA)

Complex solutions (matrix) require method of standard additions
Add small volumes higher concentration standards (change in volume is negligable)
Graph of concentration vs. absorbance
Concentration of sample is x-intercept
Overcomes problem of matrix effects

Atomic Spectroscopy for Metal Analysis

Practical Tips to Sampling
Erroneous Data and Methods of Compensation

Simple solutions (e.g. water) use standard curve technique to find unknown
concentration

Complex solutions (matrix) require method of standard additions

Add small volumes higher concentration standards (change in volume is
considered negligible)
Graph of concentration vs. absorbance
Concentration of sample is x-intercept
Overcomes problem of matrix effects

Teknik-Teknik Analisis

Ada tiga teknik yang biasa dipakai dalam analisis secara spektometri. Ketiga
teknik tersebut adalah :
Metoda Standard Tunggal
Metoda ini sangat praktis karena hanya menggunakan satu larutan standard
yang telah diketahui konsentrasinya (Cstd). Selanjutnya absorbsi larutan standard
(Astd) dan absorbsi larutsn sampel (Asmp) diukur dengan Spektrofotometri. Dari
hukum Beer diperoleh :
Astd = .b.Cstd Asmp = .b.Csmp
.b = Astd/ Cstd .b = Asmp/ Csmp
sehingga,
Astd/Cstd = Asmp/Csmp
Csmp = (Asmp/Astd) x Cstd
Dengan mengukur Absorbansi larutan sampel dan standar, konsentrasi larutan
sampel dapat dihitung.
Metode Kurva Kalibrasi
Dalam metode ini dibuat suatu seri larutan standar dengan berbagai konsentrasi
dan absorbansi dari larutan tersebut diukur dengan AAS. Langkah selanjutnya
adalah membuat grafik antara konsentrasi (C) dengan Absorbansi (A) yang akan
merupakan garis lurus melewati titik nol dengan slope = .b atau slope = a.b.
Konsentrasi larutan sampel dapat dicari setelah absorbansi larutan sampel
diukur dan diintrapolasi ke dalam kurva kalibrasi atau dimasukkan ke dalam
pers. garis lurus yang diperoleh dengan menggunakan program regresi linear
pada kurva kalibrasi

Teknik-Teknik Analisis

Metoda Adisi Standard

Metoda ini dipakai secara luas karena mampu meminimalkan kesalahan yang
disebabkan oleh perbedaan kondisi lingkungan (matriks) sampel dan standard. Dalam
metoda ini dua atau lebih sejumlah volume tertentu dari sampel dipindahkan ke dalam
labu takar. Satu larutan diencerkan sampai volume tertentu, kemudian diukur
absorbansinya tanpa ditambah dengan zat standard, sedangkan larutan yang lain
sebelum diukur absorbansinya ditambah terlebih dulu dengan sejumlah tertentu
larutan standard dan diencerkan seperti pada larutan yang pertama. Menurut hukum
Beer akan berlaku hal-hal berikut :
Ax = k.Cx;
AT = k(Cs + Cx)
dimana,
Cx = konsentrasi zat sampel
Cs = konsentrasi zat standar yang ditambahkan ke larutan sampel
Ax = Absorbansi zat sampel (tanpa penambahan zat standar)
AT = Absoebansi zat sampel + zat standar
Jika kedua persamaan diatas digabung, akan diperoleh:
Cx = Cs x {Ax/(AT-Ax)}
Konsentrasi zat dalam sampel (C x) dapat dihitung dengan mengukur Ax dan AT
dengan spektrofotometer. Jika dibuat suatu seri penambahan zat standar dapat pula
dibuat suatu grafik antara AT lawan Cs, garis lurus yang diperoleh diekstrapolasi ke AT
= 0, sehingga diperoleh:
Cx = Cs x {Ax/(0-Ax)} ; Cx = Cs x (Ax/-Ax)
Cx = Cs x (-1) atau Cx = -Cs

Kurva Kalibrasi dan Adisi Standar

Abs
Ax

AT
A=a.b.C

Cx ppm standar

-Cx

Cs-1 Cs-2 ppm tambahan

Gambar III.9 Kurva kalibrasi (kiri) dan kurva adisi standar (kanan) dalam analisis
secara spektrometri.

BEBERAPA CONTOH APLIKASI DARI AAS

Timah hitam dalam darah

Timah hitam(Pb) dalam darah dapat ditentukan dengan AAS. Mula-mula
darah ditreatment dengan Trichlorocetic Acid (TCA) untuk
mengendapkan protein, dan kemudian di centrifuge (diputar). pH dari
filtrat dibuat menjadi tiga, kemudian ditambahkan 1 ml APDC dan Pb
diekstrak dengan MIBK sebagai chelate Pb(APDC)2. Hasil ekstraksi
kemudian diukur absorbansi Pbnya dengan AAS menggunakan nyala
udara asetilen . Batas deteksi prosedur ini adalah 0,1 ppm Pb dalam
darah sedangkan kandungan normal Pbdalam darah adalah 0,3 0,4
ppm. APDC : Amonium pirolidin dithiokarbamat; MIBK : Metil Isobutil
Keton.
Seng (Zn) dalam Tumbuhan
Zn adalah unsur runut esensial dalam tumbuhan. Beberapa gram
sampel tumbuhan yang telah dikeringkan dan dan digerus diabukan
dalam krus silika selama semalam pada suhu 500 oC. Abu yang
diperoleh dilarutkan dengan 6 M HCl dan dikeringkan perlahan-lahan
dengan steambath. Endapan yang diperoleh dilarutakan ke dalam HCl
encer kemudian disaring. Selanjutnya absorbansi Zn dalam larutan ini
diukur dengan AAS.

Aplikasi AAS

Tembaga dalam air laut

Beberapa elemen dalam air laut berada dalam tingkat ppb atau lebih rendah.
Oleh karenanya harus dipekatkan terlebih dahulu sebelum dianalisis misalnya
Cu dalam air laut (1-25 ppb). Cu dalam air laut dapat ditentukan dengan AAS
setelah diekstrak sebagai kompleks APDC pada pH larutan kira-kira 3. Pelarut
ekstraksi yang dipakai adalah metil-n-amil keton yang kelarutannya dalam air
cukup rendah.
Berilium dalam partikel udara
Partikel dikumpulkan dengan kertas cellulose asetat membrane (misal e.g.,
Milipore). Filter ini kemudian diabukan pada suhu rendah. Abu yang diperoleh
dilarutkan dalam Hcl encer kemudian diukur dengan AAS. Hasil dilaporkan
dalam mg/meterkubik udara.
Na, K, Mg, dan Ca dalam semen
Sampel didekomposisi dengan 4 M HCl dan diuapkan sampai kering, endapan
yang diperoleh dilarutkan kembali dengan 4 M HCl dan diencerkan. Larutan
yang diperoleh diukur absorbansinya untuk masing unsur di atas. Gangguan
dari phosphat, silikon, dan aluminium pada pengukuran absorbansi Ca dapat
dihilangkan dengan menambahkaaaan larutan strontium konsentrasi tinggi.

Question
A series of solutions is made up by adding 0.1, 0.2, 0.3, 0.4 and 0.5 mL of a
10 mg L-1 lead standard to 100 mL aliquots of the unknown solution. The following
results were obtained:

Volume std. (mL)

Abs

0
0.27

0.1
0.37

0.2
0.53

0.3
0.65

0.4
0.75

0.5
0.88

Plot a calibration graph and determine the concentration of the unknown

Assuming constant volume of 100 mL, the concentration increase in the
5 solutions are 10, 20, 30, 40, and 50 g L-1.
(e.g. 0.5 mL of 10 mg/L = 5 x 10-3 mg in 100 mL = 5 g/0.1 L = 50 g L-1)
Absorbance = (0.01235 x conc) + 0.2694
Unknown = 21.8 g L-1 lead

Quantification