Atomic Absorption Spectroscopy

David Joram R. Mendoza Institute of Chemistry, University of the Philippines Los Baos Laguna 4031, Philippines mendozadavidjoram@yahoo.com
ABSTRACT For years, atomic absorption spectroscopy has paved the way for the analysis of elements which became very essential in various fields. In this experiment, milk and chocolate samples were utilized to quantitatively measure the concentration of Ca 2+ and Na+ using atomic absorption spectroscopy. In addition, trace lead analysis using the standard addition technique was performed to quantitatively measure the concentration of lead in a water sample from Molawin Creek using the same instrumentation. From a theoretical data given for Ca analysis, it was determined that the calcium content of the sample is estimated to be 0.2068 ppm (with La3+) and 0.1798 ppm (without La3+). However, no valid quantification of sodium content of the sample was made due to lack of time and resources. Lastly, trace lead was analyzed on water samples from Molawin Creek. The standard addition technique was used in the preparation of the solutions. Based on the calculated values, the concentrations of lead on the water samples were 0.0382 ppm, 0.2277 ppm and 0.0562 ppm for trials 1, 2 and 3, respectively. On the average, the lead content of the water samples collected is 0.1074 ppm.

Keywords: Atomic Absorption Spectroscopy, Sodium and Calcium Analyses, Trace Lead Analysis, Standard Addition Technique
1 INTRODUCTION In 1955, Allan Walsh introduced atomic absorption spectroscopy (AAS) which paved the way for the extensive and intensive quantitative determination of the trace metals in liquids. For instance, calcium, magnesium, sodium and potassium ions were quantitatively determined in blood serum. AAS was also able to quantify the amount of nickel, copper, lead, selenium and vanadium in edible oils, beer samples, gasoline, urine and lubricating oil, respectively (Kar, 2005). It was observed that a metal sample, upon transforming into gaseous form by means of a flame or electrothermal device, absorbs radiant energy related to their own specific resonance wavelength. Hence, a part of the light having the same resonance wavelength is absorbed by the sample subjected to the flame. This observation ultimately built the basis of atomic absorption spectroscopy (AAS) (Kar, 2005). At the present, Lajunen & Permmaki (2004) defined atomic absorption spectroscopy/spectrometry as an analytical method for the determination of elements in small quantities and is based on the absorption of radiation energy by free atoms. The underlying principle of AAS is the absorption of energy exclusively by ground state atoms while they are in the gaseous form. It is specifically the amount of light at the resonant wavelength absorbed that is quantified in AAS. The absorption of light is dependent on the atom being analyzed itself. Atoms absorb light at a definite wavelength depending on the nature of the elements (Khopkar, 1998). The amount of light absorbed is proportional to the number of atoms in the light path. From this relationship, a quantitative determination of the analyte present in a sample can be made. Certain special light sources and walength controllers are used in order to uniquely determine the presence of a specific element whenever there is a mixture of elements (Beaty & Kerber, 1993). Figure 1 shows a block diagram of a basic atomic absorption spectrometer. It is mainly composed of a light source, a sample cell, a light measurement unit comprised of a monochromator, detector and readout (Beaty & Kerber, 1993).

Figure 1| Block diagram of a basic atomic absorption spectrometer. One of the major advantages of using AAS for quantitative determination is its ability to facilitate selective quantification of a specific element. Hence, there is no need to separate the analyte of interest from a mixture of sample. Thus, AAS offers the conservation of time and money while also eliminating the possibility of various sources of errors incurred by other processes. Estimation of both aqueous and non-aqueous solutions is also possible with atomic absorption spectroscopy (Kar, 2005). In this experiment, milk and chocolate samples were utilized to quantitatively measure the concentration of Ca2+ and Na+ using AAS. In addition, trace lead analysis using the standard addition technique was performed to quantitatively measure the concentration of lead in a water sample from Molawin Creek.

2 METHODOLOGY

2.1 Preparation of standard solutions 2.1.1 For Ca Analysis 50 mL of 50 ppm calcium solution was initially prepared from a 1000 ppm stock solution. Using different volumes of this prepared solution, 50 mL solutions of different concentrations were made as shown in Table 1. Table 1| Amounts of standards needed to prepare calcium solutions of various concentrations. Volume of standard (mL) Concentration (ppm) 0.0 0.0 0.5 0.5 1.0 1.0 1.5 1.5 2.0 2.0 2.5 2.5 2.1.2 For Na Analysis 50 mL of 10 ppm sodium solution was initially prepared from a 1000 ppm stock solution. Using different volumes of this prepared solution, 50 mL solutions of different concentrations were made as shown in Table 2. Table 2| Amounts of standards needed to prepare sodium solutions of various concentrations. Volume of standard (mL) Concentration (ppm) 0.0 0.0 1.0 0.2 2.0 0.4 4.0 0.8 5.0 1.0 0.0 0.0 Prior to dilution with deionized water, 2.0 mL of 2% HNO3 and 5.0 mL 5% Cs+ were added. Solutions were then kept to polyethylene bottles. Meanwhile, another set of standards were prepared without the Cs+ solution. 2.1.1 For Trace Pb Analysis (Standard Addition Technique) 50 mL of 10 ppm lead solution was initially prepared from a 1000 ppm stock solution. Using different volumes of this prepared solution, 50 mL solutions of different concentrations were made as shown in Table 3. Table 3| Amounts of standards needed to prepare lead solutions of various concentrations. Volume of standard (mL) Concentration (ppm) 0.0 0.0 0.5 0.5 1.0 1.0 1.5 1.5 2.0 2.0 2.5 2.5

Prior to dilution with deionized water, 5.0 mL of the sample was added. Solutions were then kept to polyethylene bottles. 2.2 Preparation of samples 2.2.1 Milk/Chocolate Powdered milk and chocolate and liquid milk were utilized as samples. Three trials were made for each sample; two of which requires the addition of a releasing agent and the third trial having none. To an evaporating dish, 3.0 g powdered milk/chocolate or 10.0 mL liquid milk was transferred. The sample was charred/evaporated until dryness in a hot plate. These were then transferred to a furnace at 550oC for 3-4 hours and cooled to room temperature. Two milliliters of concentrated HNO3 and 10 mL distilled water were also added to the solution. After boiling and cooling, the mixture was quantitatively transferred with filtration to a 50-mL volumetric flask and then diluted with deionized water. Furthermore, 5.0 mL of the prepared samples were transferred to a 50-mL volumetric flask, added with 2.0 mL 2%HNO3 and 5.0 mL of the releasing agent (La3+ for Ca analysis and Cs+ for Na analysis). 2.2.1 Water A liter of water sample from Molawin Creek was taken. Then, 400 mL of sample was initially used and 5.0 mL concentrated HNO3 was then added. The solution was allowed to evaporate to almost 100 mL by heating in a hot plate. The rest of the water sample was then added to the solution and concentrated again until the remaining amount was around 100 mL. Additional 5.0 mL concentrated HNO3 was added and the reaction mixture was covered with watchglass. The mixture was concentrated to 50 mL, cooled and filtered. Five milliliters of this sample is added to lead standard solution for trace lead analysis. 2.3 Analysis using Atomic Absorption Spectroscopy The standards and samples were analyzed using atomic absorption spectroscopy by following the necessary procedures stated in the operating instruction manual. Calibration curves were constructed and concentrations of the elements in the samples were then estimated and compared.

3 RESULTS AND DISCUSSION Three elements (Na+, Ca2+ and trace Pb) were quantified on different samples using atomic absorption spectroscopy (AAS). Sodium and calcium were quantified on powdered/liquid milk and powdered chocolate while trace Pb was quantified on water samples from Molawin Creek. All measurements taken from the experiment were made possible by using the atomic absorption spectrometer. Just like any other absorption spectrometer, three basic parts must be present: a light source, a sample cell (3) a means of a specific light measurement (see Figure 2) ( Beaty & Kerbe, 1993).

For both types of AA spectrometer, a light source must be present in order to emit the sharp atomic lines of the element to be examined. Specifically, this light source must be a line source which emits specific the specific wavelengths which can be absorbed by the atom. Light sources vary from element to element but the most widely used are the hollow cathode lamp and electrodeless discharge lamp as shown in Figure 5 and 6, respectively ( Beaty & Kerbe, 1993).

Figure 2| Requirements of a spectrometer. Though they share some common similarities, UV-Vis spectrophotometry and AAS are two different methods of analysis. AAS use metal ions as samples while specific compounds are quantified by UV-Vis spectrophotometry. In addition, in AAS, a specific wavelength of electromagnetic radiation is provided in contrary to UV-Vis spectrophotometry that provides a range of wavelengths. Figure 1 previously showed the basic parts of a typical atomic absorption spectrometer. This can be further classified as being single-beam or double-beam based in its photometer (Beaty & Kerbe, 1993). In a single beam AA spectrometer, there is only one path for all measurements as shown in Figure 3.

Figure 5| Hollow cathode lamp (HCL)

Figure 6| Electrodeless discharge lamp (EDL). Briefly, a hollow cathode lamp consists of a glass cylinder filled with an inert gas usually Argon or Neon at low pressure. The cathode is made especially such that it is made from the same metal to be determined. The hollow cathode lamp process consists of three major stages, namely: spluttering, excitation and emission as shown in Figure 7 ( Beaty & Kerbe, 1993).

Figure 3| A single-beam AA spectrometer. On the other hand, a double-beam AA spectrometer utilizes additional optics in order to divide the light from the light source which directs to the reference cell and sample cell as shown in Figure 4 ( Beaty & Kerbe, 1993). Figure 7| Hollow cathode lamp process. Generally, some of the drawbacks of using a hollow cathode lamp include (1) shelf life of HCLs, (2) replacement of lamp for a particular element and (3) deposition of metal atoms on sides and end windows during spluttering (Beaty & Kerbe, 1993). Figure 4| Double-beam AA spectrometer. On the other hand, electrodeless discharge lamps are used when HCLs are not able to provide quality analyses due to its

limitations. Such case is the involvement of the more volatile elements where low intensity and short life are a problem. EDLs can provide a brighter and more stable source of light. Generally, electrodeless discharge lamps are said to more intense and sensitive than HCLs. Apart from superior performance, the shelf life of EDLs is longer than HCLs and thus, provides more reliable and accurate analytical analyses (Beaty & Kerbe, 1993). The light source can be controlled or modulated (that is, switched on and off rapidly) to amplify light coming from the source and at the same time, ignoring emission from the sample cell. This can be done by the use of a chopper, located between the source and the sample cell, or through pulsing of power to the source ( Beaty & Kerbe, 1993). The light from the source then passes through the sample cell. However, the sample is treated such that it is converted into gaseous phase by generally introducing the sample into a burner system or electrically heated furnace with respect to the optical path of the spectrometer ( Beaty & Kerbe, 1993). The flame process is diagrammed in Figure 8.

Figure 9| Basic atomic absorption burner system. One essential factor considered in the flame process is the flame temperature. Cooler flames are said to cause incomplete atomization due to insufficient energy and hence, analyses using these flames are likely to encounter more interference problems. Two of the most commonly used oxidant-fuel mixtures are the air-acetylene and nitrous-oxide acetylene. The air-acetylene flame is a satisfactory source of atomization of the sample but the hotter nitrous oxide-acetylene flame is more preferred when dealing with refractory-forming elements and when lessening some types of interferences. Table 4 shows temperatures for some flames commonly used in AAS (Beaty & Kerbe, 1993). Table 4| Temperatures of pre-mix flames. Oxidant-Fuel Temperature, C Air-methane 1850-1900 Air-natural gas 1700-1900 Air-hydrogen 2000-2050 Air-acetylene 2125-2400 N2O- acetylene 2600-2800 After atomizing the sample, the specific light measurement unit takes into place. A monochromator is able to disperse the wavelengths of light emitted from the sample and eventually isolates the wavelength of interest. The isolation of the particular line of interest is essential to distinguish the analyte in the presence of the others (Beaty & Kerbe, 1993). A typical monochromator is diagrammed in Figure 10.

Figure 8| Flame process in AAS (M+ - metal cation; A- associated anion) The sample is initially aspirated into the burner chamber where a mixture of fuel and oxidant gases is present. Evaporation or desolvation then happens as the metals (initially present as droplets in aerosol) pass into the heat of the flame. This process also removes the solvent and leaves tiny solid particles of sample material. As liquefaction proceeds, the sample is subjected to even more heat, totally vaporizing the sample. At this point, the analyte, M+, is still bound to some anion, A-. However, this molecule does not yet possess the atomic absorption phenomenon necessary for the analysis. As more heat is applied, this molecule eventually disintegrates into individual atoms desired for making measurements for analysis. Figure 9 shows a basic atomic absorption burner system ( Beaty & Kerbe, 1993).

Figure 10| A typical monochromator. The isolated wavelength of light then directs into the detector which is normally a photomultiplier tube. A photomultiplier tube produces an electrical current which depends on the intensity of light. The electrical current is then subjected into an amplifier producing a signal that is directly related to the light attenuation occurring in the sample cell.

Instrument readout can also be produced directly into concentration units (Beaty & Kerbe, 1993). In this experiment, liquid/powdered milk and powdered chocolate were utilized as samples. For calcium analysis, a set of standard calcium solution were prepared. The absorbances of the standards and the sample (with and without lanthanum) were then taken. However, due to erroneous results, experimental results were not considered. The erroneous results may be attributed to the use of air-acetylene mixture instead of the nitrous oxide-acetylene flame. As a consequence, chemical interferences on the analysis of calcium were not solved. Phosphate is said to be the major chemical interference in the analysis of calcium. The nitrous oxide-acetylene flame is considerably hotter than air-acetylene and can often be used to minimize chemical interferences for elements generally determined with air-acetylene (Beaty & Kerbe, 1993).Hence, a theoretical data were provided. These data is given in Table 5. Table 5| Absorbance readings of standard calcium solutions and sample. Concentration of Ca2+ Absorbance Reading (ppm) 0 0.0013 0.5 0.0988 1 0.1108 1.5 0.273 2 0.3497 2.5 0.4278 Unknown w/o La3+ 0.0239 Unknown w/ La3+ 0.0286

Calcium phosphate does not totally dissociate in air-acetylene flame. Thus, increasing the concentration of phosphate ions lead to the decrease on its absorbance. However, in the presence of lanthanum, a thermally stable compound will be formed with the interferent. That is, lanthanum will tie up the phosphate which paves the way for the atomization of the calcium. Lanthanum is referred to as a releasing agent (Beaty & Kerbe, 1993). In addition, Figure 12 shows the calibration curve for the determination of calcium in the sample.

0.45 0.4 0.35 0.3 0.25 0.2 0.15 0.1 0.05 0 0

Absorbance

y = 0.1741x - 0.0074 R = 0.9706

1 2 Ca Concentration (ppm)

Figure 12| Standard curve for the determination of calcium content of milk sample. From the standard curve, the calcium content of the sample is estimated to be 0.2068 ppm (with La3+) and 0.1798 ppm (without La3+). For sodium analysis, the intensity of emission of light was considered. Again, liquid/powdered milk and powdered chocolate were utilized as samples. Sodium standard solutions were prepared initially. The emissions of these solutions were then taken as shown in Table 6. In addition, Figure 13 shows the calibration curves generated. Table 6| Emission readings of the water samples for Na analysis. Concentration n (ppm) 0.0 0.2 0.4 0.8 1.0 Emission w/o Cs+ 0.000 0.1014 0.2925 0.5320 0.5982 w/ Cs+ 0.0000 0.0124 0.3057 0.5034 0.6578

From the data above, it can be noticed that the absorbance of the sample untreated with La3+ is lower than the treated sample. Again, this may attributed to fact that chemical interferences, such as phosphate, may ruin the atomization process of the flaming process (Beaty & Kerbe, 1993). Figure 11 shows the effect of phosphate on calcium.

Figure 11| Interference of phosphate on calcium.

addition, Figure 14 (see next page) shows the standard curves for the quantification of trace in the water samples. Table 7 | Absorbance readings of the water samples, prepared by standard addition with Pb. Concentration (ppm) 0.0 0.2 0.4 0.8 1.0 Figure 13| Calibration curves for the determination of sodium content in milk/chocolate sample. From the data gathered, it can be noticed that emission of solutions with treatment of Cs+ is higher compared to those solutions that were not treated. This is because some ionization is still experienced even the oxidant-fuel mixture is the recommended air-acetylene flame. Ionization should be suppressed by the incorporation of excess cesium ( for sodium determinations), at concentrations of 1000 g/mL or greater, in the form of chlorides or nitrates, in both sample and standard solutions. Extent of ionization is inversely proportional to analyte concentration with errors due to incomplete suppression thus being greater at lower concentrations (Cantle, 1982). Furthermore, an erroneous result on reading the emission of sample was incurred and so the sodium content was not able to be quantified. This result is probably due to the very high sodium content of the sample. Dilutions were not able to be performed due to lack of time and resources. Lastly, trace lead was quantified in water samples from Molawin creek. The preparation utilized the standard addition technique which is a useful technique to work in presence of matrix interference without eliminating the interference itself, and still make an accurate determination of analyte concentration. This is made possible by constructing the calibration curve in terms of the sample and the matrix interference. In other words, aliquots of a standard are added to the sample which permits any present interferent in the sample to also affect the standard similarly (Beaty & Kerbe, 1993). Table 7 shows the absorbances of the water samples prepared by standard addition with Pb for the three trials. In Based on the calculated values, the concentrations of lead on the water samples were 0.0382 ppm, 0.2277 ppm and 0.0562 ppm for trials 1, 2 and 3, respectively. On the average, the lead content of the water samples collected is 0.1074 ppm. Absorbance Trial 2 0.00720 0.01280 0.02290 0.02820 0.04200

Trial 1 0.0007 0.0027 0.0051 0.0101 0.0126

Trial 3 0.0011 0.0067 0.0107 0.0204 0.0255

0.0500 0.0400 Absorbance 0.0300 0.0200 0.0100 0.0000 0.0

Trial 3

Trial 2

Trial 1

y = 0.03197x + 0.00728 R = 0.94868 y = 0.02402x + 0.00135 R = 0.99881 y = 0.01204x + 0.00046 R = 0.99873 0.2 0.4 0.6 0.8 Concentration (ppm) 1.0 1.2

Figure 14 | Standard curve for the determination of lead content of water samples by standard addition.

4 CONCLUSIONS In this experiment, milk and chocolate samples were utilized to quantitatively measure the concentration of Ca2+ and Na+ using atomic absorption spectroscopy. In addition, trace lead analysis using the standard addition technique was performed to quantitatively measure the concentration of lead in a water sample from Molawin Creek using the same instrumentation. From a theoretical data given for Ca analysis, it was determined that the calcium content of the sample is estimated to be 0.2068 ppm (with La3+) and 0.1798 ppm (without La3+). In the presence of a releasing agent, lanthanum, absorption values are greatest since chemical interferences due to calcium is lessened. In addition, for Na analysis, , it was observed that emission of solutions with treatment of Cs+ is higher compared to those solutions that were not treated. This is because some ionization is still experienced even the oxidant-fuel mixture is the recommended air-acetylene flame. Ionization should be suppressed by the incorporation of excess cesium. However, no valid quantification of sodium content of the sample was made due to lack of time and resources. Lastly, trace lead was analyzed on water samples from Molawin Creek. The standard addition technique was used in the preparation of the solutions. Based on the calculated values, the concentrations of lead on the water samples were 0.0382 ppm, 0.2277 ppm and 0.0562 ppm for trials 1, 2 and 3, respectively. On the average, the lead content of the water samples collected is 0.1074 ppm.

6 REFERENCES Beaty, R & Kerber, J (1993). Concepts, Instrumentation and Techniques in Atomic Absorption Spectroscopy, 2e. CT, USA: The Perkin-Elmer Corporation 9-36 p. Cantle, J (Ed.) (1982). Atomic Absorption Spectrometry. NY, USA: Elsevier Science Publishing Company, Inc. , 171 p. Khopkar, S (1998). Basic Concepts of Analytical Chemistry, 2e. New Delhi, Indonesia: New Age International (P) Ltd., Publishers, 259 p. Kar, A (2005). Pharmaceutical Drug Analysis, 2e. New Delhi, Indonesia: New Age International (P) Ltd., Publishers, 259 p. Lajunen, L & P Permki (2004). Spectrochemical Analysis by Atomic Absorption and Emission, 2e. Cambridge, UK: The Royal Society of Chemistry, 12 p