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Tuesday 2 July 2013

ATOMIC ABSORPTION SPECTROSCOPY


read this
http://www.nuigalway.ie/chemistry/level2/courses/CH205_atomic_absorption_spectroscopy.pdf, this will give full information

Atomic absorption spectroscopy (AAS) is a spectroanalytical procedure for the quantitative determination of chemical elements employing the absorption of optical radiation (light) by free atoms in the gaseous state.
In analytical chemistry the technique is used for determining the concentration of a particular element (the analyte) in a sample to be analyzed. AAS can be used to determine over 70 different elements in solution or directly in solid samples employed in pharmacologybiophysics and toxicology research.
Atomic absorption spectrometry was first used as an analytical technique, and the underlying principles were established in the second half of the 19th century by Robert Wilhelm Bunsen and Gustav Robert Kirchhoff, both professors at the University of Heidelberg, Germany.

Modern atomic absorption spectrometers
The modern form of AAS was largely developed during the 1950s by a team of Australian chemists. They were led by Sir Alan Walsh at the CSIRO (Commonwealth Scientific and Industrial Research Organization), Division of Chemical Physics, in MelbourneAustralia.
The technique makes use of absorption spectrometry to assess the concentration of an analyte in a sample. It requires standards with known analyte content to establish the relation between the measured absorbance and the analyte concentration and relies therefore on the Beer-Lambert Law. In short, the electrons of the atoms in the atomizer can be promoted to higher orbitals (excited state) for a short period of time (nanoseconds) by absorbing a defined quantity of energy (radiation of a given wavelength). This amount of energy, i.e., wavelength, is specific to a particular electron transition in a particular element. In general, each wavelength corresponds to only one element, and the width of an absorption line is only of the order of a few picometers (pm), which gives the technique its elemental selectivity. The radiation flux without a sample and with a sample in the atomizer is measured using a detector, and the ratio between the two values (the absorbance) is converted to analyte concentration or mass using the Beer-Lambert Law.

Atomic absorption spectrometer block diagram
In order to analyze a sample for its atomic constituents, it has to be atomized. The atomizers most commonly used nowadays are flames and electrothermal (graphite tube) atomizers. The atoms should then be irradiated by optical radiation, and the radiation source could be an element-specific line radiation source or a continuum radiation source. The radiation then passes through a monochromator in order to separate the element-specific radiation from any other radiation emitted by the radiation source, which is finally measured by a detector.



The atomizers most commonly used nowadays are (spectroscopic) flames and electrothermal (graphite tube) atomizers. Other atomizers, such as glow-discharge atomization, hydride atomization, or cold-vapor atomization might be used for special purposes.


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Flame atomizers

The oldest and most commonly used atomizers in AAS are flames, principally the air-acetylene flame with a temperature of about 2300 °C and the nitrous oxide (N2O)-acetylene flame with a temperature of about 2700 °C. The latter flame, in addition, offers a more reducing environment, being ideally suited for analytes with high affinity to oxygen.

A laboratory flame photometer that uses a propane operated flame atomizer
Liquid or dissolved samples are typically used with flame atomizers. The sample solution is aspirated by a pneumatic analytical nebulizer, transformed into an aerosol, which is introduced into a spray chamber, where it is mixed with the flame gases and conditioned in a way that only the finest aerosol droplets (< 10 μm) enter the flame. This conditioning process is responsible that only about 5% of the aspirated sample solution reaches the flame, but it also guarantees a relatively high freedom from interference.
On top of the spray chamber is a burner head that produces a flame that is laterally long (usually 5–10 cm) and only a few mm deep. The radiation beam passes through this flame at its longest axis, and the flame gas flow-rates may be adjusted to produce the highest concentration of free atoms. The burner height may also be adjusted, so that the radiation beam passes through the zone of highest atom cloud density in the flame, resulting in the highest sensitivity.
The processes in a flame include the following stages:
  • Desolvation (drying) – the solvent is evaporated and the dry sample nano-particles remain;
  • Vaporization (transfer to the gaseous phase) – the solid particles are converted into gaseous molecules;
  • Atomization – the molecules are dissociated into free atoms;
  • Ionization – depending on the ionization potential of the analyte atoms and the energy available in a particular flame, atoms might be in part converted to gaseous ions.
Each of these stages includes the risk of interference in case the degree of phase transfer is different for the analyte in the calibration standard and in the sample. Ionization is generally undesirable, as it reduces the number of atoms that is available for measurement, i.e., the sensitivity. In flame AAS a steady-state signal is generated during the time period when the sample is aspirated. This technique is typically used for determinations in the mg L-1 range, and may be extended down to a few μg L-1 for some elements.

Electrothermal atomizers

Electrothermal AAS (ET AAS) using graphite tube atomizers was pioneered by Boris V. L’vov at the Saint Petersburg Polytechnical Institute, Russia, since the late 1950s, and further investigated by Hans Massmann at the Institute of Spectrochemistry and Applied Spectroscopy (ISAS) in Dortmund, Germany.
Although a wide variety of graphite tube designs have been used over the years, the dimensions nowadays are typically 20–25 mm in length and 5–6 mm inner diameter. With this technique liquid/dissolved, solid and gaseous samples may be analyzed directly. A measured volume (typically 10–50 μL) or a weighed mass (typically around 1 mg) of a solid sample are introduced into the graphite tube and subject to a temperature program. This typically consists of stages, such as:
  • Drying – the solvent is evaporated
  • Pyrolysis – the majority of the matrix constituents is removed
  • Atomization – the analyte element is released to the gaseous phase
  • Cleaning – eventual residues in the graphite tube are removed at high temperature.
 
The graphite tubes are heated via their ohmic resistance using a low-voltage high-current power supply; the temperature in the individual stages can be controlled very closely, and temperature ramps between the individual stages facilitate separation of sample components. Tubes may be heated transversely or longitudinally, where the former ones have the advantage of a more homogeneous temperature distribution over their length. The so-called Stabilized Temperature Platform Furnace (STPF) concept, proposed by Walter Slavin, based on research of Boris L’vov, makes ET AAS essentially free from interference. The major components of this concept are:
  • Atomization of the sample from a graphite platform inserted into the graphite tube (L’vov platform) instead of from the tube wall in order to delay atomization until the gas phase in the atomizer has reached a stable temperature;
  • Use of a chemical modifier in order to stabilize the analyte to a pyrolysis temperature that is sufficient to remove the majority of the matrix components;
  • Integration of the absorbance over the time of the transient absorption signal instead of using peak height absorbance for quantification.
In ET AAS a transient signal is generated, the area of which is directly proportional to the mass of analyte (not its concentration) introduced into the graphite tube. This technique has the advantage that any kind of sample, solid, liquid or gaseous, can be analyzed directly. Its sensitivity is 2–3 orders of magnitude higher than that of flame AAS, so that determinations in the low μg L-1 range (for a typical sample volume of 20 µL) and ng g-1 range (for a typical sample mass of 1 mg) can be carried out. It shows a very high degree of freedom from interferences, so that ET AAS might be considered the most robust technique available nowadays for the determination of trace elements in complex matrices.

Specialized Atomization Techniques

While flame and electrothermal vaporizers are the most common atomization techniques, several other atomization methods are utilized for specialized use.
 
Glow-Discharge Atomization
A glow-discharge (GD) device serves as a versatile source, as it can simultaneously introduce and atomize the sample. The glow discharge occurs in a low-pressure argon gas atmosphere between 1 and 10 torr. In this atmosphere lies a pair of electrodes applying a DC voltage of 250 to 1000 V to break down the argon gas into positively charged ions and electrons. These ions, under the influence of the electric field, are accelerated into the cathode surface containing the sample, bombarding the sample and causing neutral sample atom ejection through the process known as sputtering. The atomic vapor produced by this discharge is composed of ions, ground state atoms, and fraction of excited atoms. When the excited atoms relax back into their ground state, a low-intensity glow is emitted, giving the technique its name.
The requirement for samples of glow discharge atomizers is that they are electrical conductors. Consequently, atomizers are most commonly used in the analysis of metals and other conducting samples. However, with proper modifications, it can be utilized to analyze liquid samples as well as nonconducting materials by mixing them with a conductor (e.g. graphite).
 
Hydride Atomization
Hydride generation techniques are specialized in solutions of specific elements. The technique provides a means of introducing samples containing arsenic, antimony, tin, selenium, bismuth, and lead into an atomizer in the gas phase. With these elements, hydride atomization enhances detection limits by a factor of 10 to 100 compared to alternative methods. Hydride generation occurs by adding an acidified aqueous solution of the sample to a 1% aqueous solution of sodium borohydride, all of which is contained in a glass vessel. The volatile hydride generated by the reaction that occurs is swept into the atomization chamber by an inert gas, where it undergoes decomposition. This process forms an atomized form of the analyte, which can then be measured by absorption or emission spectrometry.
Cold-Vapor Atomization
The cold-vapor technique an atomization method limited to only the determination of mercury, due to it being the only metallic element to have a large enough vapor pressure at ambient temperature. Because of this, it has an important use in determining organic mercury compounds in samples and their distribution in the environment. The method initiates by converting mercury into Hg2+ by oxidation from nitric and sulfuric acids, followed by a reduction of Hg2+ with tin(II) chloride. The mercury, is then swept into a long-pass absorption tube by bubbling a stream of inert gas through the reaction mixture. The concentration is determined by measuring the absorbance of this gas at 253.7 nm. Detection limits for this technique are in the parts-per-billion range making it an excellent mercury detection atomization method.



Applications of Atomic Absorption Spectroscopy
water analysis (e.g.Ca, Mg, Fe, Si, Al, Ba content)
food analysis
analysis of animal feedstuffs (e.g.Mn, Fe, Cu, Cr, Se,Zn)
analysis of additives in lubricating oils and greases (Ba,Ca, Na, Li, Zn, Mg)
analysis of soils
clinical analysis (blood samples: whole blood, plasma,
serum; Ca, Mg, Li, Na, K, Fe
A) Sample preparation
Depending on the information required, total recoverable metals, dissolved metals, suspended metals, and total metals could be obtained from a certain environmental matrix. Table 1 lists the EPA method number for sample processing in terms of the environmental matrices and information required. For more detail information, readers could refer to EPA document SW-846 "Test methods for evaluating solid wastes".

Table 1 EPA sample processing method for metallic element analysis

Analysis Target
Method Number
Environmental Matrice
total recoverable metals
3005
ground water/surface water
dissolved metals
3005
ground water/surface water
suspended metals
3005
ground water/surface water
total metals
3010
aqueous samples, wastes that contain suspended solids and mobility-procedure extracts
total metals
3015
aqueous samples, wastes that contain suspended solids and mobility-procedure extracts
total metals
3020
aqueous samples, wastes that contain suspended solids and mobility-procedure extracts
total metals
3050
sediments, sludges and soil samples
total metals
3051
sludges, sediment, soil and oil

Appropriate acid digestion is employed in these methods. Hydrochloric acid digestion is not suitable for samples, which will be analyzed by graphite furnace atomic absorption spectroscopy because it can cause interferences during furnace atomization.

B) Calibration and standard curves
As with other analytical techniques, atomic absorption spectrometry requires careful calibration. EPA QA/QC demands calibration through several steps including interference check sample, calibration verification, calibration standards, bland control, and linear dynamic range.
The idealized calibration or standard curve is stated by Beer's law that the absorbance of an absorbing analyte is proportional to its concentration.
Unfortunately, deviations from linearity usually occur, especially as the concentration of metallic analytes increases due to various reasons, such as unabsorbed radiation, stray light, or disproportionate decomposition of molecules at high concentrations. Figure 3 shows an idealized and deviation of response curve. The curvature could be minimized, although it is impossible to be avoided completely. It is desirable to work in the linearity response range. The rule of thumb is that a minimum of five standards and a blank should be prepared in order to have sufficient information to fit the standard curve appropriately. Manufacturers should be consulted if a manual curvature correction function is available for a specific instrument.
Figure 3. Idealized/deviation response curve

If the sample concentration is too high to permit accurate analysis in linearity response range, there are three alternatives that may help bring the absorbance into the optimum working range:
1) sample dilution
2) using an alternative wavelength having a lower absorptivity
3) reducing the path length by rotating the burner hand.

C) EPA method for metal analysis
Flame atomic absorption methods are referred to as direct aspiration determinations. They are normally completed as single element analyses and are relatively free of interelement spectral interferences. For some elements, the temperature or type of flame used is critical. If flame and analytical conditions are not properly used, chemical and ionization interferences can occur.
Graphite furnace atomic absorption spectrometry replaces the flame with an electrically heated graphite furnace. The major advantage of this technique is that the detection limit can be extremely low. It is applicable for relatively clean samples, however, interferences could be a real problem. It is important for the analyst to establish a set of analytical protocol which is appropriate for the sample to be analyzed and for the information required. Table 2 lists the available method for different metal analysis provided in EPA manual SW-846.

Table 2. EPA methods for determination of metals by direct aspiration

Analyte
Method number
Analyte
Method number
Analyte
Method number
aluminum
7020
antimony
7040
barium
7080A
beryllium
7090
cadmium
7130
calcium
7140
chromium
7190
cobalt
7200
copper
7210
iron
7380
lead
7420
lithium
7430
magnesium
7450
manganese
7460
molybdenum
7480
nickel
7520
osmium
7550
potassium
7610
silver
7760A
sodium
7770
strontium
7780
thallium
7840
tin
7870
vanadium
7910
zinc
7951





D) Interferences
Since the concentration of the analyte element is considered to be proportional to the ground state atom population in the flame, any factor that affects the ground state population of the analyte element can be classified as interference. Factors that may affect the ability of the instrument to read this parameter can also be classified as interference. The following are the most common interferences:
A) Spectral interferences are due to radiation overlapping that of the light source. The interference radiation may be an emission line of another element or compound, or general background radiation from the flame, solvent, or analytical sample. This usually occurs when using organic solvents, but can also happen when determining sodium with magnesium present, iron with copper or iron with nickel.
B) Formation of compounds that do not dissociate in the flame. The most common example is the formation of calcium and strontium phosphates.
C) Ionization of the analyte reduces the signal. This is commonly happens to barium, calcium, strontium, sodium and potassium.
D) Matrix interferences due to differences between surface tension and viscosity of test solutions and standards.
E) Broadening of a spectral line, which can occur due to a number of factors. The most common line width broadening effects are:
1. Doppler effect
This effect arises because atoms will have different components of velocity along the line of observation.
2. Lorentz effect
This effect occurs as a result of the concentration of foreign atoms present in the environment of the emitting or absorbing atoms. The magnitude of the broadening varies with the pressure of the foreign gases and their physical properties.
3. Quenching effect
In a low-pressure spectral source, quenching collision can occur in flames as the result of the presence of foreign gas molecules with vibration levels very close to the excited state of the resonance line.
4. Self absorption or self-reversal effect
The atoms of the same kind as that emitting radiation will absorb maximum radiation at the center of the line than at the wings, resulting in the change of shape of the line as well as its intensity. This effect becomes serious if the vapor, which is absorbing radiation is considerably cooler than that which is emitting radiation.




recap
Atomic Absorption Spectroscopy
Introduction
Atomic absorption absorption spectroscopy (AA or AAS) is one of the commonest instrumental methods for analyzing for metals and some metalloids.
AAS
Metalloids like antimony, arsenic, selenium, and tellurium are now routinely analyzed by hydride generation AAS (HGAAS; see www.shsu.edu/~chm_tgc/sounds/sound.html and www.shsu.edu/~chemistry/primers for animations and primers on that method). Inductively coupled plasma (ICP) is also a powerful analytical, instrumental method for these elements but at this point its much higher cost limits it widespread use as compared to AAS.
As the animation on AAS here shows, the main parts of the AAS system are a hollow cathode lamp, nebulizer, air/acetylene flame, and optical system. Alternate sample introduction systems such as graphite furnaces are also available but will not be discussed here. The job of each are detailed below:
 
Job of the hollow cathode lamp
Provide the analytical light line for the element of interest
Provide a constant yet intense beam of that analytical line
Job of the nebulizer
Suck up liquid sample at a controlled rate
Create a fine aerosol for introduction into the flame
Mix the aerosol and fuel and oxidant thoroughly for introduction into the flame
Job of the flame
Destroy any analyte ions and breakdown complexes
Create atoms (the elemental form) of the element of interest
Fe0, Cu0, Zn0, etc.
Job of the monochromator
Isolate analytical lines' photons passing through the flame
Remove scattered light of other wavelengths from the flame
In doing this, only a narrow spectral line impinges on the PMT.
Job of the photomultiplier tube (PMT)
As the detector the PMT determines the intensity of photons of the analytical line exiting the monochromator.

The Hollow Cathode Lamp
The hollow cathode lamp (HCL) uses a cathode made of the element of interest with a low internal pressure of an inert gas. A low electrical current (~ 10 mA) is imposed in such a way that the metal is excited and emits a few spectral lines characteristic of that element (for instance, Cu 324.7 nm and a couple of other lines; Se 196 nm and other lines, etc.). The light is emitted directionally through the lamp's window, a window made of a glass transparent in the UV and visible wavelengths.
 
Neublizer, Different Oxidants, and Burner Heads, and Waste
AAS Nebulizer and Flame
The nebulizer chamber thoroughly mixes acetylene (the fuel) and oxidant (air or nitrous oxide), and by doing so, creates a negative pressure at the end of the small diameter, plastic nebulizer tube (not shown in adjacent figure; see figure below). This negative pressure acts to suck ("uptake") liquid sample up the tube and into the nebulizer chamber, a process called aspiration. A small glass impact bead and/or a fixed impeller inside the chamber creates a heterogeneous mixture of gases (fuel + oxidant) and suspended aerosol (finely dispersed sample). This mixture flows immediately into the burner head where it burns as a smooth, laminar flame evenly distributed along a narrow slot in the well-machined metal burner head.
Liquid sample not flowing into the flame collects on the bottom of the nebulizer chamber and flows by gravity through a waste tube to a glass waste container (remember, this is still highly acidic).
For some elements that form refractory oxides (molecules hard to break down in the flame) nitrous oxide (N2O) needs to be used instead of air (78% N2 + 21% O2) for the oxidant. In that case, a slightly different burner head with a shorter burner slot length is used.
 
The Monochromator and PMT
Tuned to a specific wavelength and with a specified slit width chosen, the monochromator isolates the hollow cathode lamp's analytical line. Since the basis for the AAS process is atomic ABSORPTION, the monochromator seeks to only allow the light not absorbed by the analyte atoms in the flame to reach the PMT. That is, before an analyte is aspirated, a measured signal is generated by the PMT as light from the HCL passes through the flame. When analyte atoms are present in the flame--while the sample is aspirated--some of that light is absorbed by those atoms (remember it is not the ionic but elemental form that absorbs). This causes a decrease in PMT signal that is proportional to the amount of analyte. This last is true inside the linear range for that element using that slit and that analytical line. The signal is therefore a decrease in measure light: atomic absorption spectroscopy.
 
Acidic Content and Oxidation State of Samples and Standards
The samples and standards are often prepared with duplicate acid concentrations to replicate the analyte's chemical matrix as closely as possible. Acid contents of 1% to 10% are common.AAS procedure
In addition, high acid concentrations help keep all dissolved ions in solution.
The oxidation state of the analyte metal or metalloid is important in AAS. For instance, AAS analysis of selenium requires the Se(IV) oxidation state (selenite). Se(VI), the more highly oxidized state of the element (selenate), responds erratically and non reproducibly in the system. Therefore, all selenium in Se calibration standards and Se containing samples must be in the Se(IV) form for analysis. This can be accomplished by oxidizing all Se in the sample to selenate using a strong oxidizer such as nitric acid or hydrogen peroxide and then reducing the contained selenate to selenite with boiling HCl.
 
Double Beam Instruments
The light from the HCL is split into two paths using a rotating mirror: one pathway passes through the flame and another around. Both beams are recombined in space so they both hit the PMT but separated in time. The beams alternate quickly back and forth along the two paths: one instant the PMT beam is split by the rotating mirror and the sample beam passes through the flame and hits the PMT. The next instance, the HCL beam passes through a hole in the mirror and passes directly to the PMT without passing through the flame. The difference between these beams is the amount of light absorbed by atoms in the flame.
The purpose of a double beam instrument is to help compensate for drift of the output of the hollow cathode lamp or PMT. If the HCL output drifts slowly the subtraction process described immediately above will correct for this because both beams will drift equally on the time scale of the analysis. Likewise if the PMT response changes the double beam arrangement take this into account.
 
Ignition, Flame conditions, and Shut Down
The process of lighting the AAS flame involves turning on first the fuel then the oxidant and then lighting the flame with the instrument's auto ignition system (a small flame or red-hot glow plug). After only a few minutes the flame is stable. Deionized water or a dilute acid solution can be aspirated between samples. An aqueous solution with the correct amount of acid and no analyte is often used as the blank.
Careful control of the fuel/air mixture is important because each element's response depends on that mix in the burning flame. Remember that the flame must breakdown the analyte's matrix and reproducibly create the elemental form of the analyte atom. Optimization is accomplished by aspirating a solution containing the element (with analyte content about that of the middle of the linear response range) and then adjusting the fuel/oxidant mix until the maximum light absorbance is achieved. Also the position of the burned head and nebulizer uptake rate are similarly "tuned." Most computer controlled systems can save variable settings so that methods for different elements can be easily saved and reloaded.
Shut down involves aspirating deionized water for a short period and then closing the fuel off first. Most modern instruments control the ignition and shutdown procedures automatically.

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