In this lecture

• Atomic absorption spectroscopy (AAS)

Atomic Absorption
– History
Spectroscopy (AAS) – General principle of AAS
– Instrumentation of AAS
Assoc. Prof. PHAM VAN HUNG – Practice problem
Department of Food Technology

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Introduction
Introduction
• Accurately measuring trace amounts of
mineral elements.
• AAS is an analytical method based on the
absorption of ultraviolet or visible radiation • Application in food analysis, nutrition,
by free atoms in the gaseous state. biochemistry, and toxicology.
• Application of plasma as excited sources
• AAS is commonly used for analysis mineral for Inductively coupled plasma - atomic
nutrients and toxicants in foods emission spectroscopy: ICP-AES
• Recently, ICP-MS are used.
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 Major minerals Trace minerals
• Inorganic atoms or molecules
• Macrominerals (7) • Microminerals or trace elements
• Needed in > 100 mg/day • < 100 mg/day needed
– Calcium – Iron
– Phosphorus – Zinc
– Magnesium – Iodine
– Sodium – selenium
– copper
– Chloride
– manganese
– Potassium – fluoride
– Sulfur – chromium
– molybdenum
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Atomic Absorption Spectroscopy:

Toxic mineral An Aussie Invention
• Developed by Alan Walsh (below) of the
• Aluminum CSIRO in early 1950s.
• Arsenic
• Cadmium
• Lead
• Mecury

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 Energy transitions in atoms Absorption and Emission
• Atomic absorption spectra are produced when
ground state atoms absorb energy from a Excited States
radiation source.
• Atomic emission spectra are produced when
excited neutral atoms emit energy on returning
to the ground state or the lower energy state.
Ground State
• The absorption of photon of radiation causes an
outer shell electron to jump to a higher energy Absorption Emission
level.
 When atoms absorb light, the incoming energy excites
• Atoms absorb or emit radiation of discrete an electron to a higher energy level.
wavelength because the allowed levels of  Electronic transitions are usually observed in the
electrons in atoms are fixed. visible or ultraviolet regions of the electromagnetic
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spectrum.

Atomization Atomization
• Atomic spectroscopy requires that atoms
• Atomization involves separating particles
of element of interest in the atomic state
into individual molecules (vaporization)
– Not combined with other elements in a and breaking molecules into atoms.
compound – Expose the analyte to high temperatures in a
– They are well separated in space flame or plasma.
– Vaporize and decompose the analyte to
• In foods, all elements are present as atoms that may absorb radiation or become
compounds or complexes. excited and subsequently emit radiation.
– Must be converted to neutral atoms

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 Atomisation Principles of AAS
The metal vapor absorbs energy from an
external light source, and electrons jump from
the ground to the excited states
MX M M+ The ratio of the transmitted to incident light
energy is directly proportional to the
Solution Solid Gas Atom Ion
concentration of metal atoms present
Excitation A calibration curve can thus be constructed
[Concentration (ppm) vs. Absorbance]
M* M+* 13 14

Atomic Absorption/Emission/
Atomic Absorption Spectroscopy Fluorescence Spectroscopy
• 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

Metal Zn Fe Cu Ca Na
λ (nm) 214 248 325 423 589
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 Methods for atomization Atomic Absorption Spectroscopy
• The analyte concentration is determined from
the amount of absorption.

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Overview of AA Sample
spectrometer. Compartment
Atomic Absorption Spectroscopy
• It is possible to measure the concentration of
an absorbing species in a sample by applying
the Beer law:

 I  Abs  cb Light Source Detector

Abs  log 
Io 

 = extinction coefficient 19 20

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Atomic Absorption Spectroscopy Radiation Sources
• Instrumentation • Hollow-Cathode Lamps (most common).
– can be used to detect one or several atomic
• Radiation Sources species simultaneously

• Atomizer
• Lasers (more specialised).
• Monochromator – more sensitive
– can detect only one element at a time
• Detector (disadvantage)

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Hollow-Cathode Lamps Hollow-Cathode Lamps

• When voltage is applied across the electrode, the

- A hollow tube filled with argon or neon. lamp emits radiation characteristic of the metal in
- An anode made of tungsten the cathode.
- a cathode made of the metallic form of the element being • HCLs for about 60 metallic elements are available.
measured 23 24

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 Atomisation Flame Atomisation
• Flame AAS can only
• Atomic Absorption Spectroscopy (AAS) requires analyze solutions.
that the analyte atoms be in the gas phase.
• A slot-type burner is used to
increase the absorption
• Vapourisation is usually performed by: path length (recall Beer
– Flames law).
– Furnaces
• Solutions are aspirated with
– Plasmas the gas flow into a
nebulising/mixing chamber
to form small droplets prior
to entering the flame.
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Flame atomization Flame Atomisation

• Three types of flames • Degree of atomisation is temperature dependent.
– Stoichiometric flame is produced from
stoichiometric amounts of oxidant and fuel so
• Vary flame temperature by fuel/oxidant mixture.
the fuel is completely burned and the oxidant
is completely consumed.
Fuel Oxidant Temperature (K)
– Oxidizing flame is produced from a fuel-lean
Acetylene Air 2,400 - 2,700
mixture. The hottest flame and a clear blue Acetylene Nitrous Oxide 2,900 - 3,100
appearance. Acetylene Oxygen 3,300 - 3,400
– Reducing flame is produced from fuel-rich Hydrogen Air 2,300 - 2,400
mixture. A cool flame and a yellow color. Hydrogen Oxygen 2,800 - 3,000
Cyanogen Oxygen 4,800
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 Furnaces Furnaces
• Improved sensitivity over flame sources.

• Less sample is required.

• Generally, the same temp range as flames.

• More difficult to use, but with operator skill at the

atomisation step, more precise measurements
can be obtained.

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Inductively Coupled Plasmas Analytical procedure

• Enables much higher temperatures to be achieved. • Sample preparation

Uses Argon gas to generate the plasma. • Standard preparation
• Temps ~ 6,000-10,000 K. • Absorbance measurement
• Used for emission spectroscopy rather than • Calculation
absorption spectroscopy due to the higher sensitivity
and elevated temperatures.
• Atoms are generated in excited states and
spontaneously emit light.

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 Wet Ashing Wet Ashing
 a procedure for oxidizing organic substances by 1. Evaporate moist samples (25–50 ml) in an appropriate
using acids and oxidizing agents or their dish at 100◦C overnight or in a microwave drying oven
combinations. until dry.
 Minerals are solubilized without volatilization. 2. Heat on a hot plate until smoking ceases.
 Its primary use is preparation for specific mineral 3. Ash in a 525◦C furnace for 3–8 h.
analysis and metallic poisons.
4. Remove dish from furnace and allow to cool. Ash
 Analytical testing laboratories use only wet ashing in
should be grayish white to white and free from carbon.
preparing samples for certain mineral analyses (e.g.,
Fe, Cu, Zn, P), because losses would occur by 5. Cool and wet with deionized distilled water plus 0.5–
volatilization during dry ashing. 3.0 ml of HNO3.

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Wet Ashing AAS - Calibration Curve

• The instrument is calibrated before use by testing the
6. Dry on a hot plate or steam bath and then return to a
absorbance with solutions of known concentration.
525◦C furnace for 1–2 h.
7. Repeat steps 5 and 6 if carbon persists. • Consider that you wanted to test the sodium content of
bottled water (A = 0.650?).
8. Dissolve the ash in 5 ml of 1M HNO3 by warming on a
hot plate for 2–3 min to aid solution. Transfer to an • The following data was collected using solutions of
appropriate size volumetric flask (i.e., 50 ml), then sodium chloride of known concentration
repeat with two additional portions of 1M HNO3.
9. The specific type of mineral being analyzed using AAS Concentration (ppm) 2 4 6 8
or ICP-MS.
Absorbance 0.18 0.38 0.52 0.76
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 Use of Calibration curve to determine sodium
Calibration Curve for Sodium concentration {sample absorbance = 0.65}

A A
b 1.0 b 1.0

s s
o 0.8
o 0.8

r r 0.6
0.6
b b
a 0.4 a 0.4

n n Concentration
c 0.2
c 0.2
Na+ = 7.3ppm
e e
2 4 6 8 2 4 6 8

Concentration (ppm) 37 Concentration (ppm) 38

Sample Problem 1 Sample problem 2

Determination of Nickel
• Calculate iron concentration in 10g of flour in 100 mL
• The nickel content in river water solution. The absorbance as follows.
was determined by AA analysis Content by AA
after 5.00 L was trapped by ion 120
exchange. Rinsing the column
with 25.0 mL of a salt solution y = 5.6x + 20
released all of the nickel and
Absorbance Units

the wash volume was adjusted 80

to 75.00 mL; 10.00 mL aliquots
of this solution were analyzed
by AA after adding a volume of 40
0.0700 g Ni/mL to each. A
plot of the results are shown
below. Determine the
concentration of the Ni in the 0
river water. 0 5 10 15
Volum e of Nickel Added(m L)

Answer: 0.375 g/L

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The end!

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