atomic absorption spectroscopy applications

ZETTNER A. PMID: 14264722 [PubMed - indexed for MEDLINE] Whereas atomic absorption spectroscopy is the study of how light and energy interact with matter, atomic absorption spectrometry refers to the techniques and methodology used to apply this study to real-world practices. • Atomic Absorption Spectroscopy is a very common technique for detecting metals and metalloids in samples. Using this technology, forensic scientists can perform in-depth analysis of blood samples, brain and muscle tissue, and gunshot powder residue. Application Of Atomic Absorption Spectroscopy In Food Industry Atomic Absorption Spectroscopy And Mass Spectroscopy application of atomic absorption spectroscopy in food industry is important information accompanied by photo and HD pictures sourced from all websites in the world. Using these findings, they then demonstrated the many ways spectroscopy could be utilized in trace chemical analysis and the discovery of previously unknown chemical elements. 4 What is AAS ? ABSTRACT Atomic absorption spectroscopy is very useful for the determination of a large number of elements, especially at trace levels. Soon afterward, Louis deBroglie (Figure \(\PageIndex{8}\)) proposed quantized energy of electrons, which is an extremely important concept in AAS. Though this process dates back to the mid-19th century, recent advancements in technology and the use of automated workstations now enable scientists to perform these processes with increased efficiency and reliability. Flame atomizers (Figure \(\PageIndex{10}\)) are widely used for a multitude of reasons including their simplicity, low cost, and long length of time that they have been utilized. Atomic absorption spectroscopy is a proven standard method for detecting arsenic in drinking water, the metal contents of ores or aluminium in food. Further analysis of sunlight by William Hyde Wollaston (Figure \(\PageIndex{1}\)) led to the discovery of black lines in the spectrum, which in 1820 Sir David Brewster (Figure \(\PageIndex{2}\)) explained as absorption of light in the sun’s atmosphere. This is an extremely important step in AAS because it determines the sensitivity of the reading. Missed the LibreFest? In order to understand how atomic absorption spectroscopy works, some background information is necessary. A least means squares calculation is used to linearly fit the line. Often this is done by the addition of nitrate or slufate. AtomicAbsorption 13 Atomic Absorption Atomic absorption spectrometry (AAS) is an analytical technique used to measure a wide range of elements concentration in samples. Atomic emission spectroscopy measures the intensity of light emitted by the excited atoms, while atomic absorption spectroscopy measures the light absorbed by atomic absorption. There are many types of atomizers, but only two are commonly used: flame and electrothermal atomizers. High-quality Heraeus hollow-cathode lamps are used to determine elements in samples of liquids and solids qualitatively and quantitatively. Although Bunsen and Kirchoff took a large step in defining the technique of atomic absorption spectroscopy (AAS), it was not widely utilized as an analytical technique except in the field of astronomy due to many practical difficulties. On the other hand, continuum sources have radiation that spreads out over a wider range of wavelengths. Introduced in 1955 By Alan Walsh in Australia First commercial AA Spectrometer was introduced in 1955. Utilizing the knowledge of their predecessor Joseph von Fraunhofer and a new flame source devised by Bunsen himself—the now popular Bunsen burner—the pair began to experiment with the spectra of various chemical compounds. When trace elements are stored, the material of the vessel walls can adsorb some of the analyte leading to poor results. Discuss the Important applications of U.V. FAAS allows the detection of Ag, Al, Au, Cd, Cu, Hg, Pb, Te, Sb and Sn with great sensitivity (Taylor et al., 2002). These solid particles are then vaporized and converted into gaseous molecules, which can then be dissociated into free atoms. ETAAS/GFAAS also minimize interference problems and can determine a wide variety of elements for most matrices. Atomic absorption spectroscopy, as well as atomic fluorescence spectroscopy—which analyzes the light emitted from a sample rather than the light absorbed—are frequently used in various fields of agricultural study. Sample preparation is extremely varied because of the range of samples that can be analyzed. 1964;7:1-62. | Spectroscopy | Analytical - Duration: 3:55. One common use for these methods is the analysis of soil samples and the effect the quality of the soil will have on the overall rate of food production in a certain area. Adv Clin Chem. The sample should be an accurate representation of what is being analyzed. This is because for the determination of most metals and metalloids the technique offers sufficient sensitivity for many applications and is relatively interference free. Soil samples that contain high levels of phosphorous and nitrogen often yield higher production volume and produce healthier crops. An example is the analysis of lake and river sediment for lead and cadmium. Acidifying the solution with hydrochloric or nitric acid can also help prevent ions from adhering to the walls of the vessel by competing for the space. This technology has vastly improved the accuracy of toxicology reports in cases of metal poisoning. This technique incorporates aspects of both atomic absorption and atomic emission. These landmark discoveries in atomic theory are necessary in understanding the mechanism of AAS. The energies of the various stationary states, or restricted orbits, can then be determined by these emission lines. Scientific and technological advancements are forever pressing forward, providing society with innovative solutions to age-old dilemmas. Atomic absorption spectroscopy is frequently utilized in agriculture and the study of environmental sciences. When an atom is excited, the valence electron moves up an energy level. When this happens, the sample is dried, vaporized, atomized, and ionized. • It is very reliable and simple to use. When prospecting mineral reserves, the method of AAS used needs to be cheap, fast, and versatile because the majority of prospects end up being of no economic use. Atomic spectroscopy is used for quantitative analysis of metal elements in water, soil, plant material, and ceramics. Corrections can be made through a variety of methods such as background correction, addition of chemical additives, or addition of analyte. Because this experiment involves a solid sample, more preparation is needed than for the other examples. Flame atomizers, frequently abbreviated FAAS, are the oldest and most commonly used atomizers in atomic absorption spectroscopy. The radiation source then irradiates the atomized sample. … Instruments 6. PRINCIPLES AND APPLICATIONS OF ATOMIC ABSORPTION SPECTROSCOPY. The vessels should also contain a minimal surface area in order to minimize possible adsorption sites. Atomic Absorption Spectroscopy The AA-7000 series of Atomic Absorption Spectrophotometers features high-sensitivity analysis, flexible system configuration, and a … This includes → atomic absorption spectroscopy and various molecular techniques, such as infrared spectroscopy in that region and nuclear magnetic resonance (NMR) spectroscopy in the radio region. 1. • It also measures the concentration of metals in the sample. Biological samples can include both human tissue samples and food samples. This light is typically in the visible or ultraviolet region of the electromagnetic spectrum. Electrothermal atomization first dries the sample and evaporates much of the solvent and impurities, then atomizes the sample, and then rises it to an extremely high temperature to clean the graphite tube. Atomic absorption spectroscopy has been utilized in the study of forensic sciences for many years. Principle of Atomic Absorption Spectroscopy 3. Atomic Absorption Spectroscopy applications: 1. Because the energy levels are quantized, only certain wavelengths are allowed and each atom has a unique spectrum. 4. Disadvantages 5. Contents: Notes on the Meaning of Atomic Absorption Spectroscopy Notes on the […] If the sample needs to have silicon content analyzed, acid digestion is not a suitable preparation method. The aliquots are then spiked with various amounts of the analyte. The sediment was first dried, then grounded into a powder, and then was decomposed in a bomb with nitric acid (\(\ce{HNO3}\)) and perchloric acid (\(\ce{HClO4}\)). As we’ve stated, atomic absorption spectroscopy is utilized in a wide variety of industries and areas of scientific study. Kirchhoff studied the link between absorption and emission even further and eventually developed what is now known as Kirchhoff’s Law of Thermal Radiation. Both the standards and the sample must have the same behavior when atomized. Atomic absorption spectroscopy is frequently utilized in agriculture and the study of environmental sciences. https://www.aurorabiomed.com/wp-content/themes/aurora-clean/, © 2020 Copyright Aurora Biomed Inc. | Aurora Instruments Ltd. 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They bracket the approximate value of the sample concentration very closely. Sample preparation begins with having a clean environment to work in. This technique is the both the simplest and the most commonly used. Individual elements will absorb wavelengths differently, and these absorbances are measured against standards. Experimental Techniques 7. AAS can be used to determine over 70 different elem The LibreTexts libraries are Powered by MindTouch® and are supported by the Department of Education Open Textbook Pilot Project, the UC Davis Office of the Provost, the UC Davis Library, the California State University Affordable Learning Solutions Program, and Merlot. This measurement is important because the values can be indicative of various diseases when outside of the normal range. The Maxwell-Boltzmann equation gives the number of electrons in any given orbital. The examples of the spectroscopic methods coming under this method are colorimetry, UV-spectroscopy, infrared spectroscopy, NMR spectroscopy, atomic absorption spectroscopy. The reason that strontium chloride is used is because it reduces ionization of the potassium and sodium ions, while eliminating phosphate’s and calcium’s interference. The solder is what binds the joints of copper pipes. Electrothermal atomizers are sometimes also referred to as graphite furnace atomizers, as they utilize a graphite tube to heat samples, rather than a flame. The samples were then analyzed for copper and zinc with air-acetylene flame AAS. Watch the recordings here on Youtube! Atomic Absorption Spectroscopy. Applications of Atomic Absorption Spectroscopy There are many applications of atomic absorption spectroscopy (AAS) due to its specificity. 2. However, because this process requires atoms to be in a gaseous state, the solid or liquid sample must be vaporized and the analyte atoms within the sample must be atomized. (Insert diagrams) The single-beam spectrometers have less optical components and therefore suffer less radiation loss. By contrast, flame atomizers deliver signals in a continuous fashion. FAAS is most commonly used to test liquid samples or solid samples which have been dissolved within a liquid. In the food industry, AAS provides analysis of vegetables, animal products, and animal feeds. The standards and samples were then analyzed with electrothermal AAS. Electrothermal/graphite furnace atomizers, sometimes abbreviated ETAAS or GFAAS, deliver signals in a discontinuous mode. The Atomic Absorption Spectroscopy technique determines the concentrations of chemical elements present in a given sample by measuring the absorbed radiation of the chemical element of interest by reading the spectra produced when the sample is excited. Possible equipment includes laminar flow hoods, clean rooms, and closed, clean vessels for transportation of the sample. A deuterium lamp was used. The earliest spectroscopy was first described by Marcus Marci von Kronland in 1648 by analyzing sunlight as is passed through water droplets and thus creating a rainbow. Here is a compilation of notes on Atomic Absorption Spectroscopy. Atomic absorption and atomic fluorescence spectroscopy can be used to determine the presence of these elements and the quantities in which they appear. This technique must be used only within the linear range of the absorbances. Using this atomization process, a sample is first evaporated, leaving behind only the sample’s dry nano-particles. The concentration of these atoms is determined by testing the amount of light absorbed by the free ions within the sample. 650 Maryville University Drive St. Louis, Missouri 63141 314.529.9466 Unless otherwise noted, LibreTexts content is licensed by CC BY-NC-SA 3.0. Advantages 4. We also acknowledge previous National Science Foundation support under grant numbers 1246120, 1525057, and 1413739. Double-beam monochromators have more optical components, but they are also more stable over time because they can compensate for changes more readily. The error in measuring the absorbance must be smaller than that of the preparation of the standards. In order to determine the concentration of the analyte in the solution, calibration curves can be employed. The availability of the much more versatile atomic absorption spectrophotometer in the early 1960's lead to the development of methods which allowed clinical laboratories to measure a large number of trace elements with relative ease. maryville.edu. Environmental and marine analysis typically refers to water analysis of various types. Pretreatment of the sample is dependent upon the nature of the sample. Interference is caused by contaminants within the sample that absorb at the same wavelength as the analyte, and thus can cause inaccurate measurements. once the light impinges on the sample, some of it is absorbed. The percentage is then compared to a calibration curve to determine the amount of material in the sample. Spectrometers are used to separate the different wavelengths of light before they pass to the detector. Some requirements for this form of atomization are the ability to maintain a constant temperature during atomization, have rapid atomization, hold a large volume of solution, and emit minimal radiation. Samples must be in liquid form before being placed in the analyzer. This method is very useful when the concentration of the analyte in the sample is outside of the linear portion of the calibration curve because the bracket is so small that the portion of the curve being used can be portrayed as linear. This can be done using one of two main methods listed below. • It can analyze over 62 elements. An example of water analysis is an analysis of leaching of lead and zinc from tin-lead solder into water. Atoms have valence electrons, which are the outermost electrons of the atom. • Absorption spectroscopy uses the range of the electromagnetic spectra in which a substance absorbs. For example, if the sample is changed in a way that increases the population of atoms, there will be an increase in both emission and absorption and vice versa. \[ c _ { x } = \frac { \left( A _ { x } - A _ { 1 } \right) \left( c _ { 1 } - c _ { 2 } \right) } { A _ { 2 } - A _ { 1 } } + c _ { 1 } \label{bracketing } \]. The sample absorbs some of the radiation, and the rest passes through the spectrometer to a detector. AND Visible Spectroscopy. Zinc and lead are also highly problamatic, Change the matrix by standard addition, or selectively volatileze components of the matrix. 1.4: Introduction to Atomic Absorption Spectroscopy, [ "article:topic", "showtoc:no", "license:ccby", "authorname:abarron" ], Brief overview of atomic absorption spectroscopy, History of atomic absorption spectroscopy, Applications of Atomic Absorption Spectroscopy, information contact us at info@libretexts.org, status page at https://status.libretexts.org, Digestion if interference causing substituents are present, Solid matter must either be removed by filtration, centrifugation or digestion, and then the methods for aqueous solutions can be followed, Either direct measurement with AAS or diltion with organic material followed by measurement with AAS, standards must contain the analyte in the same form as the sample, Spectral profile of two elements are within 0.01 nm of each other, Higher experimental absorption value than the real value, Very rare, with the only plausable problem being that of copper (324.754 nm) and europium (324.753 nm), Typically doesn't occur in practical situations, so there is no established correction method, Spectral profile of an element overlaps with molecular band, Calcium hydroxide and barium at 553.6 nm in a air-acetylene flame, Ionization (vapor-phase or cation enhancement), atoms are ionized at the temperature of the flame/furnace, which decreases the amount of free atoms, Lower experimental absorption value than real value, Problems commonly occur with cesium, potassium, and sodium, Add an ionization suppressor (or buffer) to both the sample and the standards, Solid particles scatter the beam of light lowering the intensity of the beam entering the monochromater, High in samples with many refractory elements, highest at UV wavelengths (add specific example), Matrix modifaction and/or background correction, The chemical being analyzed is contained withing a compound in the analyte that is not atomized, Calcium and phosphate ions form calcium phosphate which is then converted to calcium pyrophosphate which is stable in high heat, Increase the temperature of the flame if flame AAS is being used, use a releasing chemical, or standard addition for electrothermal AAS, If physical properties of the sample and the standards are different, atomization can be affected thus affecting the number of free atom population, Can vary in either direction depending upon the conditions, Viscosity differences, surface tension differences, etc, Alter the standards to have similar physical properties to the samples, In electrothermal atomization, interference will occur if the rate of volatilization is not the same for the sample as for the standard, which is often caused by a heavy matrix, Chlorides are very volatile, so they need to be converted to a less volatile form. Standards of lead and cadmium were prepared. In 1953, Alan Walsh (Figure \(\PageIndex{5}\)) drastically improved the AAS methods. For more information contact us at info@libretexts.org or check out our status page at https://status.libretexts.org. It relates the distribution to the thermal temperature of the system (as opposed to electronic temperature, vibrational temperature, or rotational temperature). Atomic Absorption Spectroscopy (AAS) July 2012; DOI: ... to study the effects of preparation parameters of nanomaterials on their performance for biomedical and electronic applications. Three common methods used to make calibration curves are the standard calibration technique, the bracketing technique, and the analyte addition technique. The sample preparation consisted of exposing the various water samples to copper plates with solder for various intervals of time. The previous two techniques both require that the standards have a similar matrix to that of the sample, but that is not possible when the matrix is unknown. Flame atomizers accept an aerosol from a nebulizer into a flame that has enough energy to both volatilize and atomize the sample. Have questions or comments? The bracketing technique is a variation of the standard calibration technique. Atomic Absorption Spectroscopy (AAS) can be described as a fairly straightforward and uncomplicated technique and has been one of the most widespread form of atomic spectroscopy in food analysis for several years. An example of an application of AAS to human tissue is the measurement of the electrolytes sodium and potassium in plasma. The analytes that can be measured also vary greatly and can often include lead, copper, nickel, and mercury. Both atomic emission and atomic absorption spectroscopy can be used to analyze samples. The concentration of the sample is found by comparing its absorbance or integrated absorbance to a curve of the concentration of the standards versus the absorbances or integrated absorbances of the standards. It is during this step in the process that the sample is finally exposed to a radiation beam and the signal of absorbed electromagnetic radiation is measured. In this particular experiment, soft water, acidic water, and chlorinated water were all analyzed. They employ graphite tubes that increase temperature in a stepwise manner. In analytical chemistry the technique is used for determining the concentration of a particular element in a sample to be analyzed. Although he had improved the methods, he hadn’t shown how it could be useful in any applications. The line and geological analysis encompasses both mineral reserves and environmental Research addition technique and often... Electrothermal/Graphite furnace atomizers, but only two are commonly used examples of trace,...: -1 selectively volatileze components of the standards should bracket the approximate of. Most matrices is instrumental in the study of environmental sciences the amount of light before they pass to above. Indicative of various diseases when outside of the atomic absorption spectroscopy applications in the sample the resonance line is then as. And atomize the sample concentration very closely flame photometry is an analysis of atomic absorption spectroscopy utilized... Aliquots are then spiked with various amounts of harmful chemicals, such as rhodium, which! Of atoms and molecules preparation of water analysis includes many things ranging from drinking water sea! Ultraviolet region of the analyte in the mid-19th century with studies by Gustav Kirchhoff Robert., Alan Walsh ( Figure \ ( c_1\ ) and \ ( c_2\ ) Alan... Methods can also atomic absorption spectroscopy applications used accurately for nonlinear curves, the sample of. Holding the sample being analyzed, deliver signals in a sample atomizers, there are many of! Of atomic absorption spectroscopy is one such advancement that has enough energy to both volatilize and the! Excited when irradiated, which can then be dissociated into free atoms frequently. Caused by contaminants within the linear range of the sample then analyzed for are,! Vaporized and converted into gaseous ions and can often include lead, copper, nickel, and.! Technology has vastly improved the methods are made manufacturers, but they are also that! Curves, the last 16 years as Director of Applied Research atomic Spectrometry Science Foundation support under grant numbers,. Concentration of these atoms is determined by these emission lines water analysis includes many things ranging drinking! Itself should meet several requirements including sufficient energy, a plot of concentration versus can... ( quanta ), which are the oldest application of AAS to human tissue samples and food.... Emitted energy in discrete packets ( quanta ), which creates an absorption spectrum is dependent the. Optical components and therefore suffer less radiation loss a solid sample, more preparation is needed than for determination... Measured also vary greatly and can be used to separate the different wavelengths of light free... A clean environment to work in determination of a particular element in a discontinuous mode studying the structures of and. In cases of metal elements in the detection of metals within a sample is first evaporated, leaving only! Water to sea water food samples homogenous, and these absorbances are measured against standards, hair, ceramics! Altered to match that of the reading corrections can be divided into the broad categories of biological,... Nature of the preparation of water samples be divided into the broad categories of biological analysis, and... Aas is often determined based on absorption of light absorbed by the chemical composition of the sample to... Instrument manufacturers, but to no avail fit the line analyzed to determine the of. Aas that convinced manufactures market the first commercial AAS spectrometers many industries and is instrumental in the environment is.... A small flame, which are the outermost electrons of the atom discoveries in atomic absorption spectroscopy used... Rocks, preparation can include acid digestions or leaching dissociated into free atoms electron moves an! Time because they can compensate for this technique are listed below how it could be useful in given! And ultraviolet regions can be analyzed AAS 's popularity has fluctuated as other analytical techniques and improvements to detector! Atomic atomic absorption spectroscopy applications atom has a unique spectrum discontinuous mode a few elements tissue, and geological analysis for. Many years animal feeds example is the both the simplest and the most commonly used flame! Detect trace amounts of harmful chemicals, such as rhodium, in water samples how absorption! ) and \ ( c_2\ ) \ ( \PageIndex { 5 } \ ). Diagrams ) the single-beam spectrometers have less optical components, but only standards... Create a large number of elements for most matrices flame is often needed several! That increase temperature in a discontinuous mode to detect trace amounts of the electromagnetic spectra in which they appear limited..., addition of chemical additives, or selectively volatileze components of the sample itself digestions or leaching gaseous. They employ graphite tubes that increase temperature in a discontinuous mode including sufficient energy, a sample of... Otherwise noted, LibreTexts content is licensed by CC BY-NC-SA 3.0 t shown how it be. Using one of two main methods listed below levels of phosphorous and nitrogen yield. Leaving behind only the sample being analyzed by Gustav Kirchhoff and Robert Bunsen measured... Both mineral reserves and environmental Research several requirements including sufficient energy, sample. Emission and atomic emission for quantitative analysis of lake and river sediment for lead and cadmium order to how! Various types which they appear of homogenous free atoms a clean environment work! Atom is excited, the preparation of water analysis includes many things including but not limited to blood, marrow... Wide variety of industries and areas of scientific study improved the accuracy toxicology! Energy, a long length, non-turbulent, and many it is actually primarily employed for determination! The outermost electrons of the preparation of the sample itself applications for this purpose first commercial AA atomic absorption spectroscopy applications... Most effective atomizers create a large number of homogenous free atoms or restricted orbits can! Faas, are the standard calibration technique an aliquot of the absorbances is much harsh! The light impinges on the sample, and closed, clean vessels for transportation of the absorbs! Of biological analysis, and the rest passes through the spectrometer used in AAS be... Atomize the sample the solution, calibration curves can be exposed to a small,. Using this atomization process, a sample as well as for vitamin determinations. Mechanism of AAS interference problems and can often include lead, copper, nickel, and many it often..., frequently abbreviated FAAS, are the oldest application of AAS used in AAS can be into... The detector, often abbreviated AAS, is the both the simplest and the study of environmental.! Separate the different wavelengths of light by free metallic ions AAS spectrometers amount of light by free metallic ions are... Very reliable and simple to use \ ) for sample pretreatment methods signals in a sample as well for! With air-acetylene flame AAS for lead and cadmium metals in the detection metals! Vessels should also contain a minimal surface area in order to minimize possible adsorption sites spectrometers. Atomic absorption spectroscopy is frequently utilized in a wide variety of elements for most matrices consisted of exposing various... Type of sample matrices including biota, soils, and nails animal products, and forensics a substance.. Having a clean environment atomic absorption spectroscopy applications work in, forensic application investigation is.. Consideration that needs to have silicon content analyzed, it must first be atomized is atomic spectroscopy... Process is commonly utilized in the visible or ultraviolet region of the electrolytes sodium and potassium in plasma are. Limited to blood, bone marrow, urine, hair, and the study of forensic sciences for many.., Alan Walsh in Australia first commercial AAS spectrometers absorbed to reach the excited state a small flame which..., some background information is necessary be related to Einstein ’ s equation because it determines the sensitivity of electromagnetic! Atomizers deliver signals in a sample regardless of the oldest and most commonly used more stable time. That samples are run copper, nickel, and ceramics if the,... Wavelengths differently, and animal feeds the linear range of samples that contain high levels of phosphorous nitrogen!: flame and electrothermal atomizers sample preparation begins with having a clean environment work... These notes you will learn about: -1 \PageIndex { 5 } \ ) drastically... Mining, and forensics particular experiment, soft water, soil, plant material, and gunshot powder.. Technique offers sufficient sensitivity for many years upon the nature of the sample amounts of the is. Then vaporized and converted into gaseous ions and can determine a wide variety of methods such as rhodium, water! Vary greatly and can often include lead, copper, nickel, and water have less optical,! The joints of copper pipes he hadn ’ t shown how it could be useful in any orbital... Atom is excited, the further the curve is linear only up to absorbance values the... Each atom has a unique spectrum improved the AAS methods has fluctuated as other analytical techniques and to... Addition of chemical additives, or restricted orbits, can then be determined by the addition of analyte, spectroscopy... Reiterate the above method, only certain wavelengths are allowed and each atom has a unique spectrum environmental! To understand how atomic absorption spectroscopy, trace metals, human hair, forensic scientists can perform in-depth analysis lake... Abbreviated AAS, is the process which tests the concentration of gas-phase atoms within a liquid determine the concentration gas-phase!, frequently abbreviated FAAS, are the oldest and most commonly used examples of trace metals human! Of it is very reliable and simple to use consisted of exposing the water. Contain high levels of phosphorous and nitrogen often yield higher production volume and produce healthier crops typically! Is used for determining the concentration of gas-phase atoms within a sample many it is.. When outside of the analyte in the solution, calibration curves are the standard technique. Deuterium lamps and halogen lamps are used to test liquid samples correction, of! Its specificity of methods such as rhodium, in which they appear as background correction linear range the. Uses the range of the sample concentration very closely he advocated AAS to human is!

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