Atomic Absorption Spectroscopy (AAS)

Definition
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.

Application
Determination of even small amounts of metals (lead, mercury, calcium, magnesium, etc) as follows: Environmental studies: drinking water, ocean water, soil; Food industry; Pharmaceutical industry; Biomaterials: blood, saliva, tissue; Forensics: gunpowder residue, hit and run accidents; Geology: rocks, fossils 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 pharmacology, biophysics and toxicology research.

The elements presents in the host pharmaceutical substances are determined quantitavely by atomic absorption spectroscopy, for example: Pd in carbencillin sodium Cu,pb and zn in activated charcoal Fe in ascorbic acid Ag in cisplatin Zn in copper sulphate, glucogen and insulin Ni in prazosin hydrochloride Cd and pb in zinc oxide

Comparison of Atomic and Molecular spectroscopy

MAS is usually reffered to by the type of wavelength it's adsorbing, eg infra-red adsorption spectroscopy, UV-AS, or just electromagnetic adsorption spectroscopy, so you might want to search for terms such as those... AAS measures the adsorption of the constituent atoms across a region of the EM spectrum, whilst MAS measures the adsorption of the molecule across a region of the EM spectrum.

Atomization
The process which the analyte element is released to the gaseous phase 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.

How to use
To determine the concentration of a certain metal ion in a sample, the following steps occur within an atomic absorption spectrometer:
A hollow cathode lamp, with the cathode made of the metal to be tested for, emits light of a certain frequency. The light produced by the lamp is passed through the sample to be tested vaporised in a flame. The degree of light absorption is proportional to the concentration of the metal in the sample. The intensity of the light that passes through the flame is measured by a photomultiplier tube. By comparing the intensity with that produced from a control sample containing none of the metal ions being tested for, the degree of absorption, or absorbance, can be determined. The absorbance is then compared to that of a series of diluted standard solutions in order to determine the concentration. This involves the use of a calibration graph. The standard solutions should produce a straight-line graph. The absorbance recorded for the sample being tested can be matched with a concentration using the graph.

General layout of a calibration graph

Radiation sources
We have to distinguish between line source AAS (LS AAS) and continuum source AAS (CS AAS). In classical LS AAS, as it has been proposed by Alan Walsh, the high spectral resolution required for AAS measurements is provided by the radiation source itself that emits the spectrum of the analyte in the form of lines that are narrower than the absorption lines. Continuum sources, such as deuterium lamps, are only used for background correction purposes. The advantage of this technique is that only a mediumresolution monochromator is necessary for measuring AAS; however, it has the disadvantage that usually a separate lamp is required for each element that has to be determined. In CS AAS, in contrast, a single lamp, emitting a continuum spectrum over the entire spectral range of interest is used for all elements. Obviously, a high-resolution monochromator is required for this technique, as will be discussed later.

AAS detector

 The photomultiplier tube is almost universally used as the detector type in AAS. Upon exiting the sample cell the beam of monochromatic light eventually strikes the spectrophotometers detector. The role of the detector is to convert a light signal into an electrical signal. The characteristics of an ideal detector are fast response times and a linear response over a wide range of wavelengths with low noise and high sensitivity. The type of detector found in AAS is the photomultiplier tube - the principle of operation is the emission of electrons upon exposure to radiation. The detector contains a photoemissive cathode and a series of dynodes. The number of electrons emitted from the cathode is directly proportional to the intensity of the light beam. Electrons emitted from the cathode are accelerated to the first dynode by a 90 volt potential where the electron impact dislodges several additional electrons which are accelerated to the next dynode by an additional 90V potential. After nine dynodes (each one at +90V with respect to the one before it), the number of electrons finally reaching the anode is in the order of ten million for each incident photon. The current measured at the anode collector is still proportional to the intensity of the light but it has been amplified over a million times.

Atomic absorption spectrometer block diagram

Instruments
Atomizers 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.

Flame atomizer
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. 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 m) 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 510 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 L1 range, and may be extended down to a few g L-1 for some elements.

Electrothermal atomizers
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 23 orders of magnitude higher than that of flame AAS, so that determinations in the low g L-1 range (for a typical sample volume of 20 L) 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 Hydride Atomization Cold-Vapor Atomization

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.

Schematic of an atomic-absorption experiment

Instrumentation
Light source The light source is usually a hollow-cathode lamp of the element that is being measured. Lasers are also used in research instruments. Since lasers are intense enough to excite atoms to higher energy levels, they allow AA and atomic fluorescence measurements in a single instrument. The disadvantage of these narrow-band light sources is that only one element is measurable at a time. Atomizer AA spectroscopy requires that the analyte atoms be in the gas phase. Ions or atoms in a sample must undergo desolvation and vaporization in a high-temperature source such as a flame or graphite furnace. Flame AA can only analyze solutions, while graphite furnace AA can accept solutions, slurries, or solid samples. Flame AA uses a slot type burner to increase the path length, and therefore to increase the total absorbance (see Beer-Lambert law). Sample solutions are usually aspirated with the gas flow into a nebulizing/mixing chamber to form small droplets before entering the flame. The graphite furnace has several advantages over a flame. It is a much more efficient atomizer than a flame and it can directly accept very small absolute quantities of sample. It also provides a reducing environment for easily oxidized elements. Samples are placed directly in the graphite furnace and the furnace is electrically heated in several steps to dry the sample, ash organic matter, and vaporize the analyte atoms.

Light separation and detection AA spectrometers use monochromators and detectors for uv and visible light. The main purpose of the monochromator is to isolate the absorption line from background light due to interferences. Simple dedicated AA instruments often replace the monochromator with a bandpass interference filter. Photomultiplier tubes are the most common detectors for AA spectroscopy.

Interferences in AAS
Ionization Interference Only occurs in Group I elements (Li+, Na+, K+, Ce+) with low ionization potential Ideal situation - all of your element goes to the ground state, where it absorbs maximally and consistently between standards and samples Problem situation - Ionization of ground state element in flame prevents it from going to the excited state: doesn't absorb light of appropriate wavelength. Different between standards and samples. Solution - Add an excess of an ionization suppressant to standards and sampless Must be another group I element which also ionizes e.g. CeCl @ 1,000 ppm Creates lake of free electrons e-s drive Na+ or K+ back to ground state where they absorb at right l analyte absorbs consistently at maximum extent