Exercise No.

2
ATOMIC ABSORPTION SPECTROMETRY: INSTRUMENTATION,
ABSORPTION/ EMISSION TECHNIQUES

Gubantes, Gerry Mark S.

CHEM 137.1 – 2L
2nd Semester AY 2017-2018

Date Performed: February 09, 2018

Date Submitted: February 19, 2018

Mr. Jethro Magsangkay

Laboratory Instructor
 Abstract
In this exercise, instrumentation of Atomic Absorption Spectroscopy
instrument was done. This includes the sample preparation needed
for the analysis. In addition to this, calibration curve for Ca and Cu
standards were constructed. Using standard addition technique
followed by AAS, the Cu content of three different samples namely
Cobra energy drink, drinking water, and tap water were analyzed.
Results showed that the concentration of Cu present in the
respective samples were 7.26 ppm, 4.11 ppm, and 3.45 ppm.

I. Introduction

Measurements based on light and other forms of electromagnetic radiation are widely
used in analytical chemistry. The science that deals with interactions of radiation and matter is
known as spectroscopy. Spectroscopy has played a major role in the development of the modern
atomic theory as well as has provided the tools for molecular structure elucidation and both
qualitative and quantitative determination of both inorganic and organic compounds (Harvey,
2000).

Atomic spectroscopy is a technique used for the determination of the elemental

composition of an analyte by its electromagnetic or mass spectrum. The commonly used atomic
spectroscopy techniques are Flame Atomic Absorption Spectroscopy (FAAS), Graphite Furnace
Atomic Absorption Spectroscopy (GFAAS), Inductively Coupled Plasma Optical Emission
Spectroscopy (ICP-OES), and Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) (Harris,
2006).

In FAAS, atomic absorption occurs when a ground state atom absorbs energy in the form
of light of a specific wavelength and is elevated to an excited state. In GFAAAS, the sample is
introduced into a graphite tube, which is then heated in a programmed series of steps to remove
the solvent and major matrix components and to atomize the remaining sample. On the other
hand, ICP-OES is the measurement of the light emitted by the elements in a sample introduced
into an OCP source. The measured emission intensities are then compared to the intensities of
standards of known concentration to obtain the elemental composition in the unknown sample.

Atomization is the first step in all atomic spectroscopic procedures since spectroscopic
determination of atomic species can only be performed on a gaseous medium in which the
individual atoms or elementary ions are well separated from one another. Atomization is the
process in which a sample is volatilized and decomposed in such a way to produce gas-phase
atoms and ions. Absorption of light in this analytical technique is accomplished for the "free
atoms". Atomization process, moreover, is consist of five successive stages namely mixing,
evaporation, fusion, vaporization and atomization.

There are two ways of introducing a standard in analytical process. In external standard,
reference standards of known concentration are produced together with a reference blank of zero
concentration. On the other hand, internal standard involves the addition of a known amounts of
the standard to the unknown sample wherein the change in response of the unknown sample is
recorded (Harvey, 2000). Furthermore, possible interferences could occur in analyzing sample
via AAS. These include spectral, chemical, and ionization interferences.
 This study aims to (1) study the different components of an AAS System: absorption vs
emission modes, (2) determine the effects of releasing agents and/or ionization suppressants,
and (3) familiarize the standard addition techniques for trace metal analysis.

II. MATERIALS AND METHODS

The exercise was divided into three parts: calcium analysis in Bear Brand Adult,
determination of sodium in Bear Brand Kids, and analysis of lead in a water sample. Each method
involved the use of an atomic absorption spectrometer in determining absorbances. Moreover, all
stock solutions were diluted using deionized water.
For calcium analysis, a 50-mL stock solution of 50-ppm concentration was initially
prepared from a given 1000-ppm solution. From the 50-ppm Ca stock solution, standard solutions
of 50-mL volumes with concentrations of 0, 2, 4, 6, 8, and 10 ppm were prepared. Before diluting
to volume using 1% HNO3, 1 mL of 20,000 ppm La3+ was added to each working standard. The
solutions were then dispensed in plastic containers. For the preparation of sample, the previously
ignited strawberry milk powder was first dissolved in HNO3 (1:1). The resulting mixture was
filtered, and the filtrate was diluted to 100 mL. A 1:25 dilution was then performed to obtain a
sample solution.
For sodium determination, similar procedures as that for calcium analysis were done
except that 5% Cs+ was added to each working standard instead of La3+ before diluting to 50 mL.
The solutions were also placed in plastic containers. The same method of preparation was also
done for the milk sample.
For lead analysis, two methods were carried out in determining the concentration of Pb in
a water sample provided. First, using external calibration, same procedures for the preparation of
stock and standard solutions were made except that no additional releasing agents were added.
The final solutions were stored in plastic containers. As for the standard addition technique, five
50-mL volumetric flasks were obtained in which 0, 5, 10, 15, and 20 mL of prepared 6.0 ppm
standard solution were displaced. Afterwards, 10 mL of the unknown water sample was placed in
each flask. The solutions were then diluted to volume using 1% HNO3 and were placed in plastic
containers.
` After the many preparations, the Ca, Na, and Pb solutions were subjected to SpectrAA
Atomic Absorption Spectrometer 55B for AAS analysis. Initially, the instrument was calibrated and
was allowed to stand for about 30 minutes. For the sodium solutions, the instrument was set to
emission mode, the solutions were aspirated into the equipment, and the signal for every solution
was recorded as absorbance. For the calcium and lead solutions, the instrument was set to
absorption mode, the solutions were aspirated into the equipment, and the absorbance readings
were recorded. The unknown samples were also subjected to reading in order to determine the
actual concentrations of the metals from the standard curve that would be generated from the
analysis.
III. RESULTS AND DISCUSSION

The experiment involves the instrumentation and absorption/emission techniques involved in

atomic absorption spectroscopy (AAS). The general construction of an atomic absorption
spectrometery shown schematically in Figure 2.1.

Figure 2.1. General schematic of an atomic absorption spectrometer.

(Diagram obtained from Cantle, J.E. 1982. Atomic Absorption Spectrometry.
New York: Elsevier Science Publishing Company Inc., p.16.)

The components of an atomic absorption spectrometer are the light source (A), which
emits the characteristic narrow-line spectrum of the element of interest; an absorption cell or
atom reservoir in which the atoms of the sample to be analyzed are formed by thermal molecular
dissociation, most commonly by a flame (B); a monochromator (C) for the spectral dispersion of
the light into its component wavelengths with an exit slit of variable width to permit selection and
isolation of the analytical wavelength; a photomultiplier detector (D) whose function is to convert
photons of light into an electrical signal which may be amplified (E) and eventually displayed to
the operator on the instruments readout (F) (Cantle, 1982).

The first component of the atomic absorption spectrometer is the atomizer. The atomizer
breaks sample into atoms to observe atomic spectra. The most commonly used atomizer is the
chemical flame, based upon the combination of a fuel gas (e.g., acetylene) with an oxidant (e.g.,
air or nitrous oxide). The sample solution is introduced into the flame using a nebulizer in which
the passage of the oxidant creates a partial vacuum by the venture effect and thus the sample
solution is drawn up through a capillary. Thus, an aerosol is produced having a wide variety of
droplet sizes. The parts of an atomizer are shown in Figure 2.7.
 Figure 2.7. Cross-section of premix chamber of a flame atomizer.
(Image obtained from Cantle, J.E. 1982. Atomic Absorption Spectrometry.
New York: Elsevier Science Publishing Company Inc., p. 17.)

There are several types of nebulizers and atomizers. Nebulizers may be concentric-tube
pneumatic effect nebulizers, cross-flow nebulizer, fritted disk nebulizer and Babington nebulizer.
Moreover, the following are the types of atomizers for atomic spectroscopy and their
corresponding typical atomization temperature.

Table 2.1. Types of atomizers used for atomic spectroscopy*.

Type of Atomizer Typical Atomization Temperature, °C
Flame 1700-3150
Electrothermal vaporization (ETV) 1200-3000
Inductively coupled argon plasma (ICP) 4000-6000
Direct current argon plasma (DCP) 4000-6000
Microwave-induced argon plasma (MIP) 2000-3000
Glow-discharge plasma (GD) Nonthermal
Electric arc 4000-5000
Electric spark 40,000 (?)
*Information in this table obtained from Skoog, D.A., West, D.M., James Holler, W.F., and Crouch, S.R. 2014.
Fundamentals of Analytical Chemistry, 9th ed. United States: Brooks/Cole.
 The atomization process, shown in Figure 2.8, starts with nebulization, wherein the
solution sample is transformed into fine droplets by spraying through thin nozzle or passing over
vibrating crystal. Afterwards, the droplets are heated to evaporate off solvent just leaving analyte
and other matrix compounds in a process called desolvation. Through volatilization, the solid
analyte and matrix particles are converted into the gas phase. The dissociation process then
breaks down the gaseous molecules into atoms, which are then ionized to become charged.
Finally, the charged particles from ionization are subjected to light, heat, etc. for spectra
measurement (Jeffery, et. al., 1989).

Figure 2.8. The atomization process in AAS.

(Diagram obtained from Skoog, D.A., West, D.M., James Holler, W.F., and Crouch, S.R. 2014.
Fundamentals of Analytical Chemistry, 9th ed. United States: Brooks/Cole, p. 777)

There are several factors are to be considered in choosing and optimizing flame atomizers
such as the proper alignment of the flame in the light path and the type of flame used. The ideal
flame for atomic absorption should generate the correct amount of thermal energy to dissociate
the atoms from their chemical bonds. The most commonly used flames are air-acetylene and
nitrous oxide-acetylene, which produce flame temperatures of about 2300°C and 2950°C,
respectively. These types of flames are used in the analysis of 30 elements for the air acetylene
and 66 elements for nitrous oxide-acetylene. In addition, the fuel and oxidant mixture must be
controlled to provide the proper flame conditions for the element being analyzed. This is
accomplished by a gas control system providing the precise and safe regulation which is important
if reproducible results are to be obtained (Cantle, 1982).

The most familiar type of light source is the continuum or white light source, which emits
strongly in the visible and UV part of the spectrum. However, since the absorption phenomenon
being measured is occurring over an extremely narrow part of the spectrum (0.01 nm) it would
require very high resolution to measure any significant absorption from a continuum lamp. Other
problems with using typical UV/Vis continuous light source include difficulty in seeing the decrease
in signal when atoms absorb in a small bandwidth, insignificant decrease in total signal area, and
resultant bad sensitivity with large amounts of elements. A line source differs from a continuum
source in that it emits only at discrete wavelengths. The spectral lines are narrower than the
absorption lines being measured, thus high resolution is not required (Cantle, 1982).
 In AAS, there are some essential requirements in choosing light sources. It is important
that the light source emit a steady uniform level of radiation to obtain an analytical signal of low
noise level allowing good precision of measurement and good detection limits. In many cases it
is also important that the source should show little long-term drift in emission intensity. The
spectral lamp used should also require the minimum of maintenance and adjustment to obtain
optimum performance. The characteristics of the spectra produced by sharp line sources are also
important. They should be clean, with the minimum interference to the analytical line due to the
presence of spectral lines of filler gas or impurities in the lamp (Kirkbright and Sargent, 1974).

The most widely used line source for the production of fine line spectra is the hollow-
cathode lamp (HCL), which was introduced in 1916 by Paschen. It is highly advantageous due to
several factors: (1) spectral lines may be produced over the whole optical region from the infrared
to the vacuum ultraviolet; (2) intense spectra may be excited using only low currents; (3)
extremely sharp lines are quite readily obtained; (4) the lamps and their associated power
supplies are relatively simple and inexpensive to construct; and (5) they give stable, noise-free
output over long periods of time (Kirkbright and Sargent, 1974).

In general terms the hollow-cathode lamp consists of a sealed tube containing an anode,
a cylindrical hollow cathode made of, or sometimes lined with, the metal whose spectrum is
required, and an inert gas at a pressure of about 1 to 5 torr (depending on the gas used) .

Figure 2.9. A modern design of hollow-cathode lamp.

(Image obtained from Kirkbright, G.F. and Sargent, M. 1974. Atomic Absorption and
Fluorescence Spectroscopy. London: Academic Press Inc. Ltd, p. 106)

When a potential of 500 to 1000 volts is applied between the electrodes a discharge of the carrier
gas strikes and appears as a glowing positive column. However, at the pressure used, the
discharge concentrates into the hollow cathode, giving higher current densities than those
normally attained in the discharge and hence intense illumination of the cathode space. As the
ions bombard the inner surface of the cathode, a strong sputtering action ensues as they displace
atoms form the surface. The sputtered metal atoms accumulate inside the cathode and are
excited to give an emission spectrum by second-order collisions with the high concentration of
excited inert gas atoms and ions. The sputtering mechanism of the HCL explains its ability to
produce intense metal spectra even when cooled to temperatures at which the vapor pressure of
the metal is negligibly small. However, use of hollow-cathode lamps can be expensive, since
different lamps are to be used for each element being tested (Kirkbright and Sargent, 1974).

Monochromators in AAS select transmitted light from the element of interest, and not from
the incident light. They separate, isolate, and control the intensity of radiant energy reaching the
detector. Specifically, it selects a specific, narrow region of the spectrum for transmission to the
detector and rejects all wavelengths outside this region. Ideally, the monochromator should be
capable of isolating the resonance line only and excluding all other wavelengths. Several methods
may be used to isolate the required part of the spectrum, but the diffraction grating
monochromator is now universally used in atomic absorption instruments. A schematic of a
diffraction grating monochromator is shown in Figure 2.10.

Figure 2.10. Schematic of a monochromator as used in an atomic absorption spectrometer.

(Diagram obtained from Cantle, J.E. 1982. Atomic Absorption Spectrometry.
New York: Elsevier Science Publishing Company Inc., p. 31)

The radiation from the hollow-cathode lamp enters the monochromator through the entrance slit
and is focused onto the grating. The grating disperses the radiation into individual wavelengths.
By rotating the grating, the analytical wavelength of interest will pass through the exit slit and be
focused on the detector (Cantle, 1982).

Detectors for AAS/AFS do not employ photographic technique, for such is inconvenient for
measuring fluorescence signals for analytical use and would be quite impracticable for routine
absorption analysis. AAS detectors are able to convert the light signal into an electrical signal for
direct reading. Such devices must be sufficiently compact for use in the optical systems of small,
modern spectrometers, give sufficient sensitivity to produce stable and noise-free signals, have a
reasonably uniform spectral response over the wavelength range of interest and be simple to use
and give reliable operation over long periods (Kirkbright and Sargent, 1974).

Solid state photocells are the simplest of the photoelectric radiation detectors and are
widely used for light-metering applications. The mechanism of their photoconductivity is
synonymous to that of semi-conductors – the electrons may be categorized according to the band
of energy values in which they lie. When energy is supplied to the material, some electrons in the
valence band are excited into the conduction band. As the electrons in the conduction band are
more mobile this process increases the conductivity of the material. On the other hand, photo-
emissive cells depend upon the fact that when many metals are irradiated by photons of sufficient
energy, electrons are emitted from the surface of the metal (Kirkbright and Sargent, 1974).

The most commonly used detector for AAS are photomultiplier tubes. These devices
represent a logical development of the photo-emissive cell. Photons from the radiation source
bombard a cathode containing a photo-emissive substance. This causes electrons to be dislodged
from the cathode which then travel to the anode. The photomultiplier consists of a photo-emissive
cathode and an anode to collect the displaced electrons. Between the cathode and anode are
additional photo-emissive plates called dynodes. Each dynode collects electrons from the cathode
or previous dynode. The bombarding electrons dislodge several electrons from the next dynode
producing an extremely high flow of electrons at the anode. The operator controls the voltage
between anode and cathode and thus sets the ‘gain’ of the detector. This voltage will vary from
about 200-1000 volts to produce a wide range of gain settings. Since random processes within
the tube will be expanded also, the lowest voltage that is practical should always be used to avoid
excessive noise (Cantle, 1982).
Figure 2.11. Schematic representation of a photomultiplier tube.
(Diagram obtained from Kirkbright, G.F. and Sargent, M. 1974. Atomic Absorption
and Fluorescence Spectroscopy. London: Academic Press Inc. Ltd, p. 394)

The readout system in AAS presents the results from the spectroscopic process digitally.

This avoids errors in scale readings on meters trough parallax, misinterpolation between scale
divisions etc. It also consists of a logarithmic amplifier, which converts the amplified output from
the photomultiplier into read in absorbance units, the logarithmic function of percent absorption.
The scale expansion control of the spectrometer can continuously expand very small signals. This
facilitates the reading of small absorbances but it should also be accounted that any fluctuations
in the signal will be scale-expanded as well. It is not always immediately grasped that readout
directly in concentration units is achieved via the scale expand function (Cantle, 1982).

Limits of detection for flame atomic absorption spectroscopy (FAAS) are typically in the
range 0.01 to 0.1 μg mL-1. The linear dynamic range i.e, the maximum range over which the
calibration curve is linear, is limited by the propensity for self-absorption in the flame, and is
generally more than three orders of magnitude. Errors in AAS are usually minimized by the use
of a line source and the ratio method (i.e., ratio of incident to transmitted light, I0/I). Thus, if the
wavelength setting is seriously incorrect, it is unlikely that any absorption will be observed. If the
wavelength is incorrectly tuned, the effects on the value of I will roughly equal those on the value
of I0, and the error may not be too serious. A stable uptake rate or aspiration rate is required to
be taken in AAS in order for the ratio method to take effect, for such method cancels out many
instrumental errors such as long-term source drift and small monochromator drifts. The said rate
declines as the viscosity of the solution sprayed is increased. Nebulizer uptake interferences can
be minimized if the dissolved salt content of samples and standards is approximately matched
(Ebdon, et. al., 1998).

Interference is defined as an effect causing a systematic deviation in the measurement of

the signal when a sample is nebulized, as compared with the measure that would be obtained for
a solution of equal analyte concentration in the same solvent, but in the absence of concomitants.
The interference may be a result of a single or of several concomitant/s. Interference only causes
an error if not adequately corrected for during analysis. Interferences in atomic spectroscopy can
be categorized as either additive (spectral) or multiplicative (physical or chemical). Spectral
interferences result from the overlap of analyte signal with signals due to other elements or
molecules in the sample or with signals due to the flame or furnace. This type of interference is
linked with a blank or additive interference, which produces an effect that is independent of the
analyte concentration. These effects could be remedied through the preparation and analysis of
a perfect blank under the same conditions. Spectral interferences can also be removed through
background correction methods such as beam chopping, continuum source signal deduction from
line source signal, and the Zeeman effect background correction system (Ebdon, et. al., 1998;
Skoog, et al., 2014).

Analyte or multiplicative interferences change the magnitude of the analyte signal itself.
They can either be physical or chemical in nature. Physical interferences can alter the aspiration,
nebulization, desolvation, or volatilization processes. Substances in the sample that change the
solution viscosity, for example, can alter the flow rate and the efficiency of the nebulization
process. Combustible constituents, such as organic solvents, can change the atomizer
temperature and thus affect the atomization efficiency indirectly (Skoog, et. al., 2014).

Chemical interferences are usually specific to particular analytes. They occur in the
conversion of the solid or molten particle after desolvation into free atoms or elementary ions.
One cause of such interference is the formation of compounds of low volatility. The best known
of this type of interference is that of phosphate (also sulfate and silicate) on calcium, in which
the signal produced by calcium through AAS is decreased due to the decrease in the amount of
analyte atomized. This interference can be avoided by increasing the temperature of the flame
to enhance atom production, adjust the nebulizer to produce a smaller particle size, and make
observations higher in the flame. Moreover, a releasing agent, or an element that will enter into
a law of mass action competition with the analyte to combine with the interferent, is used. If an
excess of the releasing agent is added, the analyte is released from the interfering anion. Another
solution is to use a protective chelating agent which preferentially complexes the analyte,
protecting it from the grasp of the interferent. For example, excess EDTA protects calcium from
phosphate interference by binding to Ca2+ (Ebdon, et. al., 1998).

The formation of analyte oxides and hydroxides may also cause chemical interference.
Such compounds are non-volatile and are capable of absorbing intense amount of light. This can
be avoided by increasing the flame temperature and using less concentration of the oxidant.
Another chemical interferences are ionization interferences, also called vapor-phase interferences
or cation enhancement. A demonstration of this type is the doubling of the intensity of rubidium
absorption by the addition of potassium, in the air-acetylene flame. This is caused by ionization
suppression, and if uncorrected will lead to substantial positive errors when the samples contain
easily ionized elements and the standards do not. The problem is easily overcome by adding an
ionization suppressant (or buffer) in large amount to all samples and standards. The suppressant
creates high concentration of electrons, which suppresses the metal cation or analyte by shifting
equilibrium (Ebdon, et. al., 1998).

After investigating the parts and components of an atomic absorbance spectrometer, as

well as the interferences in AAS, the experiment proper was carried out. Initially, the amount of
calcium in a strawberry milk sample was identified. Calcium is of major significance for the animal
and plant kingdoms. Compounds of calcium are major components of bones and shells, and are
required for physiological functions including cell division and muscle contraction. It was one of
the first elements to be determined by flame atomic absorption spectroscopy (FAAS). Some early
works by Willis and David between 1959 and 1961 dealt with the determination of Ca in serum
and urine, and in plants and soil, respectively (Welz and Sperling, 1999).