ca122-938 SHIMADZU APPLICATION HOW TO TREAT WASTE WATER, INDUSTRIAL MATERIALS AND BIOLOGICAL MATERIALS IN LATEST ATOMIC ABSORPTION ANALYSIS - SHIMADZU CORPORATION INTERNATIONAL MARKETTING DIV. 2300-03501~10005EC-ECONTENTS: 1. Introduction. ...++ 2. Treatment of Samples......- 2-1 Treatment of inorganic substances. 2-2 Treatment of organic substances. 2-3 Solvent extraction method. 3. Atomic Absorption Analysis for Environmental Samples..... 3-1 Water analysis.......eeseeeeees 3-2 Analysis of metals contained in soil ... 4. Atomic Absorption Analysis for Metal Samples G-l Steel......-+ 4-2 Nonferrous metals. 5. Atomic Absorption Analysis for Foods......- 5-1 Sampling. .---:0++ 5-2 Treatment the sample... 6. Atomic Absorption Analysis of Plants, 6-1 Sampling..---.eeeeees 6-2 Treating the sample.. 7. Atomic Absorption Analysis of Biological Samples.........++++ 7-1 Blood and urine........- 8. Atomic Absorption Analysis of Chemical Industrial Samples.. 8-1 Petroleum. 8-2 Ceramic materials....... 8-3 Composite high molecular compounds, 9. Conclusion.! Fy Introduction The atomic absorption analysis which was originated by Dr. A. Walsh, Australia in 1955 has been employed for 30 years. During this period, the effectiveness of the analysis has been sub- stantiated and devices for practical use have been developed one after another, thereby this method being rapidly popularized in the spectro- scopic analysis fields. Presently, the atomic absorption analysis has established a stable grounding in the field of microanalysis, and it has been regarded as one of the most important means without which analytical chemistry could not been discussed in the field of the microanalysis. Such an evaluation will denote that the atomic absorption analysis possesses the features, @ that it is highly sensitive, @ that the analytical accuracy is excellent, @ that it is relatively less interfered, thereby allowing accurate analyses to be done, ® that the analytical operation is easy, and @) that the devices are compact and inexpensive, thereby extremely adaptable for the field of micro~ analysis. Treatment of samples 2-1 Treatment of inorganic substances An ordinary method to dissolve a solid sample possessing in- organic matrix is that to dissolve by means of acid. Those acids which are applied to solution of metals, alloys, and ores include HCL, H2S0,, HNOs, H3PO4, and HC20,. Some metal samples may employ a mixture of H3P0, and H2S0y. HF may be mixed with other acids to produce a stable fluoride complex which help solution of metals, or to produce Zr which is helpful to keep a sample in a state of solu- tion. Melting methods may be applied to those samples which cannot be decomposed by acid with ease. Matrices which require the melting methods contain $i0z at a high content, thereby care having to be exercised not to allow contamination by inorganic ions. 2-2 Treatment of organic substances Ordinary methods to decompose organic substances contained in samples include dry or wet ashing methods. Dry ashing method> To obtain ash of a sample, the sample should be placed in an appropriate flast and be heated at 500°C in a muffle furnace. Solid components are dried and then ashed, while the solution is evaporated. The dry ashing method is easy to use and is capable of treating a large volume of samples, while contamination is less probable. Substances to be measured may be retained on the flask wall or be hardly ashed among some samples. When a silica plate is used, especially, Co, Cu, Ag, A%, Mn, and Fe may be retained on the flask wall. When heated over 500°C, Si, AB, Co, Be, Fe, Nb, Ta, and Sn may produce acid-insoluble compounds.| i 2-3 There are some problems in the dry ashing methods, if elements to be measured are volatile. A part of or all of Pb, Hg, Cd, As, and Se may be lost in the dry ashing. Bi, Cu, Cr, Fe, Ni, Zn, and V may also be lost in the dry ashing method, since they are evapo- rated as metals, chloride, or organic metallic compouhds under some measuring conditions. Voltatile organic complex of Cu, Ni, and Fe and vanadium porphyrin compounds contained in petroleum may be evap~ orated and lost at lower temperature. < Wet ashing method > When exposed to a wet ashing method, organic substances may be oxidized and decomposed at a lower temperature than to a dry ashing method. Oxidizing agents include HNO3, HC2, HC20,, and H2S0y which are used separately or in mixture. Since the process is effected at a lover temperature, loss of elements to be measured as a result of evaporation is reduced. However, it is unavoidable that such ele~ ments as Hg, Se, and As may be lost. Fear for contamination by im- purities contained in reagents is larger when compared with a dry ashing method. A method to employ acidic mixture containing Mo(V1) or (V) as a catalyst is effective to wet-ashing of As and Se, mini~ mizing loss of volatile elements. Exercise care to handle the solvents in the atomic absorption analysis, because H2S0, often reduces the measuring sensitivity ex- tremely and HNO; and HC20, positively or negatively interfere with some elements. Solvent extraction method By extracting an objective element in a solvent which does not mix with water, there will appear so-called organic solvent effect in which more sample is introduced towards the flame. This method is helpful to condensate an objective element when the concentration of the objective element in the sample solution is too low to be ap~ plied to an atomic absorption analysis. Since the atomic absorption analysis is spectroscopically selective, solvent extraction which is not selective may be applied. When an aqueous solution of a sample contains a large volume of matrix elements or other chemical species which may generate chemical and/or physical interference, however the problem will be the degree of selectivity of a solvent extraction method to eliminate such effects. Chelate extraction system or ion association extraction system may be used for solvent extraction. In chelate extraction systems which are normally employed, pli of an aqueous solution of a sample is adjust- ed to maximize formation of extracting chelates. After feeding a che~ late reagent, it is fully mixed with an organic solvent to separate both phases. Then, the organic solvent containing metallic chelate of the objective element is sprayed over the flame for measurement. Such organic solvents should be selected as are hardly ‘dissolved with water and do not produce soot or toxic gases while burning. Benzene, toluene, and other aromatic solvents will produce sooty flame and affect the measuring accuracy. Those solvents which con~ tain chlorine such as CHC2s and CCR ate not employed because they produce disgusting toxic combustibles (hosgen). ahaGenerally ester and ketone are often employed in air-acetylene flame, since they are stable in the state of flame and are complete ly burnt, not allowing background asborbance of the flame to be ef- fected. ' In addition to them, those solvents which are often em- ployed include methyl isobutyl ketone (MIBK), ethyl acetate, iso- butyl acetate, n-butyl acetate, amyl acetate, and methyl ethyl ketone (MEK). Chelate reagents which are often used in atomic absorption anal- ysis are sodium diethyl dithiocarbamic acid (NaDDC) and ammonium pyrrolidine dithiocarbamic acid (APDC), either of which reacts with many metallic ions and produces metal chelates that can be extracted, thereby being extracted in organic solvents. In the production pro- cess of NaDDC and APDC, Pb, Cu, Zn and other metals may be mixed to form impurities. These reagents should be cleaned by means of MIBK prior to application so that those metals which compose the chelate compounds should be extracted in the MIBK. Table 1 reveals the maximum pH for extracting main elements: ea — Table 1 pH range of metal chelates for solvent extraction (MIBK) Elements| NaDC_| _APoc | Oxine [cupferron he emit = = = : af = 5-6 a6 | re fecicyy fa = 7 1 be | = 6 = | Bi esuccucry | r~1occnel) i Cd 8~1(CIICL) | 1~6 6 - co fac a 56 7 i ce [as a9 eS = \ cu fas ons a6 7 | Ga = 4 - - \ m fo 20 = = re fae as ¢ 7 4 nig amiucucyy |2~« = - ' po fac aims ¢ = 1 Mn 36 ama - 7 me | at reves | ni [ae aa 6 ' _ Le = : se fecucry face = ‘ Sb - 37 - - Sn - 3~G(CHCH,) 2.5~6 mt femucuciy [3-10 = = vy fecueny = : am [as ¢ 7 -3-aaa re ee Table 2.1 Organic reagents for solvent extraction and elements to be extracted APDC (ammonium pyrrolidine dithiocarbamic acid) H2C~CH2 S - NH N-c¢ Ha CH2 s When arranging reacting metals in the form of the periodic table: Y Cr Mn Fe Co Ni Cu Zn As Se Nb Mo Ru Rh Pd Ag Cd sn Sb Te w Os Ir Pt Au Hg TL Pb Bi u DDTC (Sodium diethyl dithiocarbamic acid) aC - CH, S~ Na HC - CH yg Hac - Cita s When arranging reacting metals in the form of the periodic table: Ti V. Cr Wn Fe Co Ni Cu Zn GA As So Mo Pd Ag Gd In Sn Sb Te W Au Hg TL Pb: Bi : u Dz or Diz (dithizone) NH - NH ~ Cells sc . NON ~ Celts When arranging reacting metals in the form of the periodic table: Ma Fe Co Ni Cu Zn Pd Ag Cd In Sn Te Pt Au Hg Tl Pb Bi Po -4-Table 2.2 Comparison among APDC, DDTC, and Dz as extractive reagents APDC (ammonium pyrrolidine dithiocarbamic acid) pote (sodium diethyl dithiocarbamic acid) Dz (dithizone) Extraction is possible in acidity (pH = 4 or more). Extraction is possible at pH S or more. Normally pH 9 or more, but extraction is feasible between pH 5 and 6. The types of elements that can be extracted are many. The types of elements that can be extracted are less than those by APDC. The types of elements that can be extracted are less than those by DDTC. MIBK solution, can be used as it is and the MIBK solution can be used as it is and the Back extraction into acid is: required, since 3) sensitivity is high. | sensitivity is high. the reagent is a solution of CHCL; or CCly. When using APDC as When using DDIC as MIBK) When back extraction 1s MIBK solution, the solution, the same effected, the standard same treatment must | treatment must be solution may be substituted a] de effected to the effected to the standard| with an aqueous solution standard solution solution side; namely, | with the same acidity with sides namely, MIBK MIBK solution has be to | the sample. solution has to be employed in this side. employed. ‘ Since the extraction . | Since the extraction | The end point of the solution is colorless, | solution is colorless, | extraction is selected vhen slit is rather difficult | it is rather difficult | the extraction solution to determine the end | to determine the end | returns to green. point of extraction. | point of extraction. Since metal complex | Metal complex solves | Metal complex is dissolved g[dissolves less in the | more when compared with | so much in CHCl) and CCl, MIBK, the solution is | APDC. that separation is complete| apt to become turbid. The reagent is so The reagent is more The reagent is as stable jjenstable that it is | stable when compared | as DoTC. decomposed at high with APDC. temperature. 8| Expensive Relatively inexpensive | Inexpensive Ordinary grade The reagent is highly |The reagent is highly available in the pure. pure. g|market is low in purity and often contains Fe, Pb , and This reagent is The reagent is easily | Since operators have been easily contaminated | contaminated during accustomed to this reagent Hoeven during operation. | operation. over a long period, they may feel at ease in operating it.3. Atomic absorption analysis for environmental samples 3-1 Water analysis How to collect a sample is very important in water analysis. With reference, especially, to quality of waste water, changes by time are supposed to occur: If the quality of waste water is found to vary in a day, the water may be collected a few times a day and be mixed to produce a sample. This method should be based on the premise that no chemical reaction might occur when mixed. A sample flask should be cleaned with acid in advance and be rinsed with the water to be collected by five times prior to collecting it as a sample. The sample flask should be sealed to prevent external substances from entering and each com- ponent in the sample from evading. But, the sealant should not be made of rubber or cork. The sample flask may be made of glass or polyethylene, but it is probable that some of glass components may be eluted in the former and that heavy metals may be adsorbed in some cases of the latter. It is desirable that a sample thus collected should be measured promptly. Should immediate measurement not be available, store it after adjusting pH to 1.0 by adding acid. Since waste water may not be measured as it is in many cases, appropriate pretreatment is often required. Should precipitates or deposits be observed in the sample, it should be pipetted only after it is fully shaked to uni- formity. Organic substances should be oxidized and decomposed by using appropriate oxidizers while turbidity should be removed by filtration or similar methods. 1) Boiling by using hydrochloric acid or nitric acid This should be applied to samples which contain an extremely small amount of organic substances and suspensions. Add 5 mf of nitric acid or hydrochloric acid per 100 mf sample, which should be heated to be boiled for about 10 minutes. 2) Decomposing by using hydrochloric acid or nitric acid This should be applied to those samples which contain hydroxide, oxide, sulfide, phosphate, and so on. After fully mix the sample, a fixed amount should be pipetted, 5 m of hydrochloride or nitric acid should be added at the rate of 100mt of a sample, and it should be concentrated to 15 m2 through heating. 3) Decomposing by using nitric acid and perchloric acid This should be applied to those samples which contains organic substance hard to be oxidized. After fully mixing the sample, a fixed volume should be pipetted, 5. 10mt of nitric acid should be added, the sample should be sub- jected to heating to decompose organic substances easily decomposed. Then, perchloric acid should be added while the nitric acid coexists for that organic substances may be treated to decompose organic sub stances. Examples of quantitative analyses of microvolumes of cadmium, zinc, lead, and copper in water are shown in Figs. 1 and 2.a I EA Tospect and collect water, 500’ me. Add 10 af of hydrochloric acid. in separatory funnel after g for S minutes. ‘Add 10 wf of ammonium hydrogen citrate. ‘Add 2 wf of hydroxylamine hydrochoride. ‘Add 2 = 3 drops of thymol blue (0.1 W/V 2) Neutvalize by adding enmonia water (I> 1) until the Liquid becones blue. Add 5 mt of ammonia vater (1 +1) Chelating? Add 10 al of dithizone chloroform (0.08 W/V 2) and mix well by shaking strongly for 2 minutes. Should the color of dithizone is changed, repeat chelation. Koacar ton vpnee layer (water phate) Place lover layer (dithizone chloroform phase) Discard funnel. Combine dithizone chloroform phases obtained by repeating chelation. Cleaning: Add 20 mE of ammonia vater (1100) and shake it co mix well: Cpacavioy Uepee layer (water phase) Discard Place the lover layer (dithizone chloroform phase) in the other separatory funnel. Cleaning: ‘Add 20 at of water and shake well. Upper layer (water phase) Bepnet og cat Place the lover layer (dithizone chloroform phase) in the other separatory funnel Back extractio ‘Add 20 mt of hydrochloric acid (2 + 100) and mix vell by shaking strongly for 2 minutes. Repeat a cycle of back extraction. Lover layer (dithizone chloroform phase) <> Combine the upper Layer (water phase) of the USE for analyzing evo back extractions in a 50 mt graduated flask, COPPER: Marking: Add pure water to accurate 50 mt in coral. Fig. 1 Measurement of Cadmiun, Zinc, and CopperWhen sampling such uniform substances displaying discontinuous com position as soil, the sampling should be effected at random from as many points as possible to minimize deviations anong analytical values. It is reasonable to effect random sampling and prepare a \ composite sample so that analysis may be carried out as to average | | i Dithizone chloroform phase after extracting cadmium, zinc, and lead F Place 100 mf in a beaker. i i Heating { evaporate chloroform } Decomposition ‘Add 2 mf of nitric acid and 2 m& of perchloric acid to decompose organic substance, and then evaporate and dry the residual materials. | E i [Dissolution ‘Add accurate 10 mf of hydrochloric acid (1 + 11) to dissolve \ them. ene y meee Weasurenent i | Fig.2 Measurement of copper | | | 3+2 Analysis of metals contained in soil i 3-2-1 Collection of samples | a) Collecting the soil values for a certain area. Since it is necessary to take vertical deviations in addition to horizontal ones into consideration, samples should be collected at a fixed interval along the section from the surface to an objective material (at least one meter deep) within a location for collection and be put to analysis. b) Collecting samples of river bottom Industrial waste water and mining debris are transported and depos~ ited on river bottoms in a form of aqueous solution, suspensions, and soild and sand, and they may be further transported downstreams, causing environmental contamination in many cases. When collecting samples, it is necessary to take the flowing direc~ tion and speed of the stream, status of the river bottom, and par~ ticle size of soils. It may be said that the objectives of sam~ pling and analyzing river bottom materials are to obtain data on volumes and distribution of heavy metals in them as potential sources of contamination.3-2-2 Pretreatment of samples a) Preparing samples After-mixing and crushing the soil collected into pieces, place it on an enameled vat and spread it uniformly. Put it to natural seasoning for one or two weeks at a well venti- lated location free from direct sunlight. It may be dried by a desiccator. When treating samples of river bottom soil or others which lots of water, moisture should be eliminated through decanta~ tion, centrifugal separation, or filtration so that precipitates may not flow away, and then it should be dried with natural wind. Should there be no time allowance for natural seasoning, adopt an appropriate analytical method applicable to characteristics of the sample. Weigh a certain volume of soil sample, dry it in a drying apparatus (105 ¥ 110°C) for 4 to 5 hours to obtain a constant volume, and calculate water content from the difference between these volumes. When treating samples which contain much moisture like river bottom soil, eliminate moisture through a centrifugal separation or a filtration method so that they may be treated in the same manner as the land soil. By sorting the dried sample by means of a 2 mm-diameter sieve to obtain a sample composed of small parti- cle of 2 mm or less in diameter. Place small-particle sample in an agate mortar and crush it into the size of 100 mesh so that an ap~ propriate analytical sample may be produced. b) Decomposing the sample 1) When obtaining the whole volume of an objective component © Fusion methods by using sodium carbonate or potassium carbonate © Decomposing method by using nitric acid/perchloric acid/hydroflu- oric acid Decomposing method by using nitric acid/sulfuric acid/hydrofluoric acid 2) When finding volumes of materials supplied and. precipitated secondarily Decomposing method by using aqua regia Decomposing method by using aqua regia/perchloric acid Strong hydrochloric acid extracting method 3) When find absorbable volumes for the purpose of grasping influence upon agricultural products © Components soluble in 0.2N hydrochloric acid © Components soluble in 12 citric acid © Components soluble in 1% acetic acid Figure 3 reveals a typical example of analyzing copper contained in soil.25.35% 402 —2tsrem 5.60% 4 ~2240m 0.28313} = sem 75% = se s.05x {ih 200" 50% HR 2m Exanple of neasurenent Caltbratton eveve of copper ot copper : Fig. 3. Example of Analyzing Copper in Soild 4. Atomic absorption analysis for metal samples 4-1 Steel 4-1-1 Sampling When sampling steel, remove scale, ferrous oxides, and other impurities adhering to the surface. Then, apply a drilling machine vertically to drill it at several points so that mean composition of the steel may be represented by the sample as much as possible. Collect small pieces whose thickness should be 1 mm or less. Samples may be collected from specific areas alone de~ pending on the objectives of analysis. If it is hard to obtain small pieces, drilling dust may be employed. After cleaning small pieces or dust, if necessary, with pure al- cohol or either to eliminate fats, collect samples for measurement. 4-1-2 Treating the samples Dissolve the samples thus collected with appropriate acid. Select such a type of acid as will not produce a product which is insoluble with the objective element, not volatize the objective element, and less interfere with it. Acids which are usually em- ployed for decomposing steel are as follows: a) To decompose it by means of HC2 and add H202 to oxidize Fe (this is used to measure At, Cr, Si, Co, Ti, Cu, Ni, and.Mn). b) To decompose it by means of HCL, and add HNO} to oxidize Fe (this is used to measure Sb and Ca). ~10-€) To decompose it by means of aqua regia (this is used to measure Af, Cr, Ti, Cu, Ni, and Bi). 4) To decompose it by means of aqua regia and smoke it wiht HCLO, (this is used to measure Zn, Ca, Co, Pb, V, Bi, Mg and Mo). e) To decompose by means of mixed acids (W12S04i3, H3P04i3, H20i16) and add HNO; to oxidize Fe (this is used to measure samples containing W). In either of these methods, filtrate impurities. When analyzing Af or Ti, melt residues by using K2S207. Examples of analyzing sluminum and cobalt in steel are illustrated in Figs. 4 and 5. Value potnead by he Gpnegnecatton of (ppm? i Taseple of wpeverenert CrtTbratton curve of alwmtows | of Mostoun in steel : Fig. 4 Analysis of Aluminum in Steel —Weigh 1,000g of a sample and place it in a beaker (200mt). Cover the beaker with a watch glass, add 20mf of hydrochloric acid (1 + 1) and gent- ly heat it to decomposition of the sample. Then, add 5 m& hydrogen per- oxide water to it to oxidize steel and subsequently boil it for about one minute to decompose hydrogen peroxide water in excess. Use a filter paper (Class 5B) to filtrate it and rinse it with warm hydrochloric acid (2 + 100) until yellow color of ferric chloride dis- appears on the filter paper. Then, rinse it with warm water a few times and keep the mixture of the filtrate and rinsing liquid as principal liq~ uid. Place residue in a platinum crucible together with the filter paper, and ash them with high heat after drying them well. Then, wet it with sulfuric acid (1 + 1), add about 3 mf of hydrofloric acid, and heat the solution to volatilize silicon dioxide and sulfuric acid (this process is not applicable to analysis of aluminum). After cooling the solution, add 1g potassium pyrosulfate and heat and melt it into a dark red solution and use the principal liquid to dissolve the molten materials. Rinse the crucible with a small amount of water to make the total volume of 100 mt which is to be used as test solution. Valve potaced ty the roo Sepegmeencton of faye) Example of measuceneat Calibration conve oF cobalt Fig. 5 Analysis of Cobalt in Steel a2Weigh 0.500g of a sample (or 1.000g of copper or cobalt) and place it in a 300 m& beaker which is in turn covered with a watch glass, add hydro- chloric acid (1 + 1) by 20 m& and gently heat and decompose the sample. Carefully add 5 mf of hydrogen peroxide water little by little to oxidize Fe, and subsequently boil it for about 1 minute to decompose hydrogen peroxide water in excess. Use a filter paper (Class 5A) to filtrate it and rinse it with warm hydrochloric acid (2 + 100) until yellow color of ferric chloride disappears on the filter paper. Then, rinse it with warm water a few times. After combining the filtrate and the rinsing liquid in a 100 m& graduated flask and cooling it to normal temperature, dilute it to a marked line with pure water to treat it as test solution. If the anticipated measuring range is exceeded, separate the sample into a certain volume of subsamples to treat them as test solution. For the calibration curve, treat pure iron in the same manner as the san- ple stated above, then add a standard solution of the element stepwise, and finally dilute it with deionized water (whose volume is equivalent to the measuring range). 4-2 Nonferrous metals 4-2-1 Sampling Nonferrous metals are diversified and complicated in types and shapes. Some metals may be crushed; they should be crushed by a crusher and then further crushed into minute particles. Shave or cut with a scissors those samples which are malleable. When shav- ing a sample with an iron or steel tool, use a strong magnet to remove iron dust mixed in the shavings. Clean fats adhered to the sample with a few percent of NaOH solution and rinse ic with water and finally with alcohol or either before drying. 4-2-2 Treating the sample Treatment may differ according to types of samples, but the following treatment is applicable to general cases. a) Copper alloy samples (not containing Sn and Si) should be heated and decomposed after adding HNO; (1 + 1) or mixed acids (HNO33: HC22: W205). If they contain Sn, dissolve the sample in a mixture of HBr and Brz (9 + 1) to volatize Sn and add HNOs and heat the mixed solu- tion to decompose bromide. If they contain lots of Si, place a sample in a platinum crucible to decompose it with HNO; (1 + 1). Then add HF and H2S0,, and heat and smoke the mixture. b) Lead alloy samples (not containing Sn) should be decomposed by using HNO; (1 + 2). I£ Sn is contained, mixed acids (NCL 85 m2; HCO,5m2; H2010m2) should be used to decompose it. c) Aluminum alloy should be decomposed with ICR. Then, add W202 and heat the sample. - 13 -rig ented Wy Te Concentration of Fe (ppm) Example of mensucenent Calibration curve of Fe or Fe in copper alley 4) Zirconium alloy should be placed in a platinum crucible. Then, add 5 mf of H20 and RF to decompose the sample. Add a few drops of HNO; to dissolve Cu. Finally add 5 m2 of H2S0, and heat and white smoke the sample. A typical example of analyzing Fe contained in copper alloy is shown in Fig. 6: For an example of analyzing aluminum alloy, refer to the Application Data AA Spectro 6. r 7 | Fig. 6 Analysis of Fe in Copper Alloy t t - leThe sample-preparing method described in JIS H1291 indicates that 1.00g of a sample should be weighed and be placed in a 200 m2 beaker vhich should in turn be covered with a watch glass, followed by addi- tion of 20 m2 of mixed acids (hydrochloric acid 1+ nitric acid 1 + water 2) and gentle heating to decompose the sample. Rinse the internal walls of the watch glass and the beaker with about 10 m& of water, and the sample should be heated gradually to expel nitrogen oxide. After cooling it, the residue should be rinsed with water into a 100 mf graduated flask and be diluted to the marked line. The diluted solution should be used as an original sample solution. This solution should be separated in accordance with the content of an element to be put to quantitative analysis so that the quantitative xange of the atomic absorption unit may be met, thereby the separated solution being used as test solution. To obtain a calibration curve, weigh 10.0g of refined copper, add 80 m nitric acid (1 + 1), and gradually heat it to decompose the components. After cooling it, add 50 mf hydrochloric acid (1 + 1) and dilute it with water to 500 mt. Separate this solution, add the stan dard solution of the element to be put to quantitative analysis, and dilute it with water to assure that ‘the measuring range of the instru- ment may be satisfied before analyzing it. 5. Atomic absorption analysis of foods 5-1 Sampling The variety of foods is so wide and they differ in shapes, ' texture, and composition. : Distribution and concentration of metal elements considerably differ among portions of the foods. Mix rice, wheat, and other cereals well so that a sample may repre~ sent the whole part. Sample 0.2 ~ 0.5kg of such cereals by re~ duction method and keep them in a polyethylene flask. Prior to starting measurements, remove an alien, sieve the sample with a plas~ tic sieve, and remove dust and sand. If dried and conditioned to the moisture content of 14 15%, gene~ rally speaking, the cereals will be free from influences from external factors, unless the temperature rises excessively. If the moisture content exceeds 25%, leave the cereals in a room to ary. After drying, the sample should be crushed and dried at 135°C for 1 hour, thereby being a sample for measurement. When sampling beans, weigh several hundreds grams and mix them well. After crushing with a crusher, dry at 130°C for an hour, thereby being a sample for measurement. For vegetables, dry a unified sam- ple at 60°C under 4,600 Pa until a constant volume is reached. For fruits, crush and unify a sample, or use a plastic grate to grate hard fruits. For fish and shell in which the texturés differ bet- ween the internal organs and meat, select a sample according to the objective of analysis. Mix a sampled material (approx. 5 v 10g) well, and dry it at 100°C for 4 v5 hours. as5-2 1) 2) 3) 4) Treating the sample Ash the sample by a dry or wet method, except special samples. Acid decomposition methods under the wet method include: To decompose with HNO; + HCLOy To decompose with HNOs + H202 To decompose with H2S0q + H202 To decompose with H2S0y + HNOs When carrying out a dry ashing method, a sample to which Mg(NOs)2 is added may be heated to protect metal atoms in the sample from being volatilized. As a typical example of analysis, a quantitative analysis of cad~ mium in rice is described hereunder: Place an appropriate volume ranging from 10 ~ 30g of a sample in a Kjeldahl flask, add 10 ~ 40 m& of deionized water and 40 mt HNO3, and mix them well. Gradually heat the mixture. Then, leave it to cool down, add 20 m2 R2S04 (95% or more), and continue heating the mixture while adding a small amount of HNOs as required, until a Light yellow or achromatic transparent solution is obtained. After cooling it, add deionized water to measure accurate 100 mt, thereby the solution being a sample. Place a fixed amount of the sample solution in a separatory funnel, add 5 mi potassium sodium tartrate, and 2 droplets of Bromophenol Blue solution as an indicator, and then add a diluted ammonia water until the color of the solution changes from light yellow to bluish purple. Finally, add deionized water to 100 mt. Then, ‘add 10 mf of (NHy)2S0,, 5 m& NaDDC solution, and 10 m& of MIBK after leaving the mixture with the former two additions for a few minutes, and then strongly shake the mixture for about 5 minutes. After leaving it to settle down, separate the MIBK phase and spray it in the air-acetylene flame, measure absorbance at the wavelength of 228.8 nm to quantitatively analyze the concentration of Cd. For further applied examples of foods analyses, refer to Application Data AA Spectro 1 and 5. 6. Atomic absorption analysis of plants 6-1 1) Sampling Dry plant samples with wind to powder and mix it well before taking a sample. Dry the sample thus collected at 85°C for four hours and in a desiccator containing silicagel for 10 days or more. Treating the sample Apply a dry or wet ashing method to the sample. The wet ashing method is effected in. the following manner: To decompose with the mixture of INO) and HCLOy. Place 0.5g of the sample in a Kjeldahl flask, add 20 m& mixed acids (HNO35 + HC£0,1), and heat the solution to evaporate and dry the residue. Continue heating to concentrate the sample until the solu- tion reaches about 2 m2, Then, add deionized water to a fixed volume. a6 SI2) To decompose with the mixture of HNO;-HF-H2S0y and HC20y. Add 25 me of the mixed acids (HNOs2 + HF2 + HzS041) to 1.0g of the sample, and heat it until white smoke appears. HCROy and heat it to complete oxidization. deionized rice leaf water. Example of me Ge'tn rice Tent and cule Fe'ln thee leat and cule - 7 Example of Analyzing Ca in Rice After cooling it, add 1 me of Then, dilute it with Figures 7 and 8 reveal analyses of Ca and Fe in and culm as a typical example. pointed by the co zo Concentration of + (ope) ‘Calibration curve of Ca Leaf and Culm Sampte oinced by the Sonate, coun oaeato oT Concentration af Fe (rpm) Cnlthration curve of Fe Fig. 8 Example of Analyzing Fe in Rice Leaf and Culm ~~7. Atomic absorption analysis of biological samples 7-1 Blood and urine Pad » 2) 3) Sampling To prevent contamination by metals, collect blood by using a disposable polyethylene injector instead of a glass one. It is recommended to employ a vaccum blood collector to measure microcomponents of elements in the blood. When utilizing plasma after collecting the blood, use heparin as anticoagulant. If a sample is not used for analysis immediately after sampling, store it by freezing at -20°C. After collecting a sample of urine store it in a dark, cold place by adding 10 m2/£ of HCL as quickly as possible. Use a pyrex flask, if possible. Treating the samples Serum samples may be sprayed directly to flame by diluting it with deionized water, but protein may be separated in some cases. Add an equal amount of acetate trichloride (8% W/V) to the serum and mix them fully. After leaving the mixture intact for 5 minutes, centrifugally separate it. Use the supernatant for mea~ suring purposes. Exercise care, however, that some elements are so strongly bonded to protein that they may not be collected well. Ashing may be sometimes employed. If an ordinary dry ashing method is applied, the sample of blood or urine may produce air bubbles or tar-like products, thereby resulting in incomplete ashing in some cases The following acid decomposition methods are applied to a wet ashing metho To decompose with HNO3. This is applied to analysis of serum. To decompose with HNO3 and HC20,. To decompose with mixed acids of HNOs, H250,, and HC20,(3 + 1+ 1). Figures 9 and 10 reveal typical medsurements of Hn and Zn con~ tained in bile. For examples of analysis of biological samples, refer to the Application Data AA Spectro 4. ~ 18 -4 Value potted by oe Yeseaee Concentration st tn (ppm) Example of messucenent of calibration cure Hovde Bile by GFA Fig. 9 Example of Measuring Mn in Bile Tale, gotneed By he ‘Calibration curve Fig. 10 Example of Measuring Zn in Bile ~19 ~ tn opm) 1 in8. Atomic absorption analysis of chemical industrial. samples 8-1 Petroleum Almost all the petroleum samples can be measured by diluting with appropriate organic solvents which include but are not limited to MIBK, P-xylene, and n-heptane. It is said that isooctane is an ideal solvent to be added to Ba, Ca, and Zn. By employing a mixture of solvents (cyclohexanone 5 + butanol 1 + ethanol 2) with RC% added at the ratio of 20 : 1, the difference in presence styles of a metal in the sample and the standard solution will not affect the measurement results. Ig a sample contains some precipitates, ashing should be effected before measurement. In this case, place 10 v 20g of the sample in a platinum crucible and heat it to ash. Place the residue in a Muffle ‘furnace and heat it at 550 v 600°C to completely oxidize car “bon. After dissolving the ash with a small amount of HC or aqua regia, take a fixed volume as a measuring sample. Figures 11 and 12 reveal typical examples of analyzing V and Ni in heavy oil. A sample was diluted by 20 times by adding methyl isobutylketone. This is an example using a sample almost similar to the standard solution and applying a calibration curve method based on a graphite furnace atomic absorption system with a pyrolytic tube. Value goinced by the oa, Concentration af ¥ (ena) lon curve of ¥ Fig. 11 Example of Analyzing V in Heavy Oil - 20 -A sample was diluted by 20 times by adding methyl isobutylketone. This is an example using a sample almost similar to the standard solution and applying a calibration curve method based on a graphite furance atomic absorption system with a pyrolytic tube. 8-2 ata fer cade es maa eta polneed by he ‘S fuample of menturenent 54 m curve of ¥ SER in heney ait Fig. 12 Example of Analyzing Ni in Heavy O11 Ceramic materials After crushing glass, cement, and other ceramic samples in an appropriate mortar and then powder them in an agate mortar. Since soda lime glass is apt to absorb carbonic acid gas or water during the crushing process, it should be crushed to 200 tm. After drying at about 110°C, decompose the sample and put it to anal- sis. Use HF for decomposing glass samples, and use HF-HC20,(4 + 5), HE-HNO3(3 + 2), or’ HF-H2SOu(1 + 1) for treatment. For further examples, refer to the Application Data AA Spectro 2. Composite high molecular compounds When applied to a dry ashing method, composite high molecular compounds start burning suddenly. After fully burning themselves, use an electric furnace for better analysis. Should flux be added, loss might occur due to the interaction between the flux and the flask and production of insoluble oxides. Polypropyrene may be ashed with more ease if NazC03 is added. Add a small amount of Na2COsas a solution to a sample placed in a platinum crucible and heat the solu~ tion to evaporate water. Cause Na2C03 to adhere to the surface of the sample and heat the sample to fuse it, and adjust the heating temperature so that the sample may be burnt with a small flame. ~ 2 =After completely burnt down, burn the ash to oxidize carbon at the temperature not to allow NazC03 to fuse, and final raise the tem- perature to fuse it. A small amount of residual carbon is completely decomposed during the fusion process, and inorganic elements are con- verted into soluble states, thereby a sample solution being obtained an acidity after adding HC2 to acidify it. 9. Conclusion Thanks to the development of the atomic absorption analysis, super~ micro analysis to the ppm ~ ppb order has been realized as a part of routine analyses. Since this method is applicable to a wide variety of elements, it is employed over the vast fields from environmental analyses to other ones as described above. It is expected that the atomic absorption analysis will further devel~ oped in the future because it is adaptable to micro analyses with much versatility. oneAluminum a) b) ce) Sample treatment Reagent Standard original solution of aluminum (1000 ppm Ag) (see item 24.1) Standard solution of aluminum (100 ppm At) Dilute the solution a) above 10 times with water. Standard solution of potassium (1000 ppm K) (See item 24.13) Processing Inspecting and sampling, 50mt te Filtration (When turbid or deposited) paces Measurement If Fe and/or Ca are contained in the sample, add a fixed amount of potassium solution (1000 ppm) to obtain the concentration of potassium of about 100 ppm. This is effected only to protect from ionizing interference. Fig. 13. Aluminum ~ 23 =Calcium Sample treatment Reagent 4) Standard original solution of calcium (1000 ppm, Ca) ») Standard solution of calcium (100 ppm Ca) Dilute the solution a) above 10 times with deionized water. ¢) Lanthanum chloride solution (5000 ppm La) Processing Process the sample in the following manner as to the analysis of calcium: Sampling, 50 me {Filtration (remove turbidity or ideposite, if any) [ Place in a 100 me messflask Add 20 mg lanthanum chloride solution. Align to the marked line. (Add deionized water to accurate 100 m2.) 00 mp) _____ Measurement] The above processing makes the solution diluted two times. Use this diluted sample solution to measure concentration of calcium and multiply it by 2 to obtain that of the original sample. Lanthanum chloride solution which is added to eliminated interference often contain calcium as impurities. In such a sase, the measured value will be larger than the actual one, resulting in inaccurate quantitative analysis, To eliminate such an error, prepare a diluted solution in which 20 m& lanthanum chloride is diluted to the total volume of 100 mt, and use this solution to effect a blank test. Should the scale be exceeded in a meter when the volume of calcium is large in the liquid thus processed, reduce the volume of the solution to be analyzed. To obtain a calibration curve, dilute the standard solution of calcium with water. Be sure to add lanthanum chloride solution at the diluting ratio of 1 : 5. Fig. 14 Ca ~ 24APDC-MIBK extraction method of cadmium and lead [Sampling, 100 mi ‘Add 10 mt of hydrochloric acid. Place in a separatory funnel after boiling for 5 minutes. Add a few drops of methyl orange solution (0.1 W/VZ). Neutralization Add ammonia (1 + 1) until the color of the liquid is neutralized to orange. Add 5 ml of acetic acid/sodium acetate buffer. (pH4-5) Quelation ‘Add 5 m& of ABDC solution (1%) and 20 mt of MIBK, and strongly shake them for 2 minutes to mix well. Leave the solution stationary for 15 minutes. e ees lower layer (water phase) a Upper layer (MIBK phase) [Measurement Fig. 15 Cadmium and lead B25 ICobalt Sample treatment Reagents Prepare the following reagents: a) Standard solution of cobalt (1000 ppm Co) b) Standard solution of cobalt (10 ppm Co): Dilute the solution a) above with deionized water by 10 times. c) Ammonium citrate solution (10 W/VZ) 4) Hydroxylamine hydrochloride solution (10 W/VZ) e) Thymol Blue solution (0.1% W/V) £) Ammonia water (1 + 1) g) Dithizone chloroform solution (0.03 W/VZ) Extraction If the concentration of cobalt is excessively low, use the following method to extract and condense it. Sampling, 500 mg Add 5 mg hydrochloric acid. Boiling For 5 minutes. After cooling, place in a l-liter separatory funnel. Add 10 mg ammonium citrate solution. Add 2 mg Hydroxylamine hydrochloride solution. “Add a few drops of Thymol Blue solution. Neutralization Add ammonia water (1 + 1) until the liquid becomes blues as | result of neutralization. <-Add another 5 mg of ammonia water (1 + 1). Quelation Add 20 mg dithizone chloroform solution and shake strongly for 2 minutes to mix Repeat quelation process again by adding 5 mg of dithizone chloroform solution. upper layer (water phase) upper layer (water phase) Discard Fig. 16 (1) Cobalt 26* Place the lower layer (dithizone phase) into a 50 m& beaker. Combine the dithizone phase obtained by repeating quelating process. Humidification Fully volatilize chloroform. Decomposition Add 2 mf nitric acid and 2 mf perchloric acid, and heat and decompose dithizone, finally evaporating and solidifying it. Leave the residue to cooling. Dissolution Dissolve the residue by adding hydrochloric acid (1 + 10) by 10 mg. ———— ‘Aligning to the marked line Add water to accurate 50 mf in total. Measurement Note: When quelating with dithizone, the extracting capability of the dithizone may be degraded in some cases where the color of dithizone is changed from green to yellow or red. In such a case, repeat quelation until the color of the dithizone does not change any longer. Fig. 16 (2) Cobalt ticsChromium Since chromium exists in water in trivalent and hexavalent states, total chromium is analyzed by combining both trivalent and hexavalent chromium or only hexavalent chromium may be analyzed. Total chromium Sample treatment Reagents a) Standard original solution of chromium (1000 ppm Cr) b) Standard solution of chromium (100 ppm Cr) Dilute the solution a) with deionized water by 10 times. ¢) Ferrous ammonium sulfate solution 4) Ammonia water (1 + 4) e) Potassium permanganate solution £) Ammonium nitrate (1 W/VZ) Processing If both trivalent and hexavalent chromium coexist, atomic absorption sensitivity may differ between these two ions. Trivalent chromium may be colloidal in water, which may produce some errors in some cases. Therefore, trivalent chromium should be oxidized into hexavalent one in the pretreatment before analysis. There are two types of pretreatments. a) When the concentration is relatively high: Sampling, 50 m& to be placed in a 100 m& beaker I Add 2 me sulfuric acid. Oxidization ‘Add potassium permanganate solution while heating until the pink color remains. Aligning the marked line Add water to 100 m& in total. Measurement The above processing makes the solution diluted by two times. Use this diluted sample solution to measure concentration of chromium and multiply it by 2 to obtain that in the original sample. Fig. 17 (1) Chromium 25 =xy S b) When the concentration is very small (this is specified in JIS KO102): fampling, 500 m& Add 2 mf of sulfuric acid (1 + 1) and boil the mixture. [Reduction (from hexavalent to trivalent) |Add 1 mg of ferrous ammonium sulfate solution. Add 2 mf of nitric acid and boil the mixture for a while (bivalnet iron to trivalent iron). Add ammonia water (1 + 4) to change into a slightly alkaline solution. Coprecipitation After boiling to remove the smell of ammonia, leave the solution at a warm place to maturate the precipitates of iron hydroxide and chrome (III). Precipitation Rinse with warm ammonium nitrate (1 W/VZ) two times (by pouring it from above the filter paper). Dissolution Dissolve with hydrochloric acid (Should the reside be hard (+ 2) by 5 mg. to be dissolved, add a small amount of warm hydrogen Place in a 100 mg massflask. POP osG0 solution.) Oxidization (trivalent to hexavalent chromium) Add potassium permanganate solution until the pink color remains, L Aligning the marked Line Add deionized water to 100 mg in total. ——_l Measurement The above processing makes the solution condensated by five times at the final stage, and use this sample to measure concentration of chromium and divide it by 5 to botain that in the original sample. Fig, 17 (2) Chromium = 29 -—s Toe oa sneninnaieniy Chromium(hexavalent) cr (V1) Sample treatment Reagents a) Standard original solution of chromium (1000 ppm Cr) b) Ferrous ammonium sulfate solution c) Ammonia water (1 + 4) Processing a) If a sample does not contain trivalent chromium: b) No special processing is required in this case. Sampling, 50 me Filtration (if the sample is turbid or deposited) Measurement If a sample contains trivalent chromium: In this case, remove trivalent chromium and iron by coprecipitating them, and use the filtrate for analysis. Sampling, 50 me Add 1 m of ferrous ammonium sulfate solution. Add ‘ammonia water (1 + 4) to change into a slightly alkaline solution. Coprecipitation After boiling to remove the smell of ammonia, leave the solution at a warm place to maturate the precipitates. discard Place the filtrate in a 100 mf beaker. Add hydrochloric acid to change into a slightly acidic solution. Aligning the marked line Add deionized water to 100 mf in total. Measurement Fig.17 (3) Chromium 30