164 Analytical Atomic Spectroscopy 4.8 BODY TISSUES AND FLUIDS 4.8.1 Introduction Analysts should constantly strive to establish maximum confidence in the results they obtain for the minimum of time expended. One important need in such an approach is that for refererice materials. To establish accurate data, samples should ideally be analysed by as many techniques as possible. By the proposed method, precision and accuracy measure- ments should be carried out on reference materials, which should then be included regularly in routine sample batches in order to benefit from the previovsly acquired data.There is a continuing need to produce and certify sufficient quantities of RMs to make a range of samples available in the routine laboratory and it is disappointing that such limited use is made of these materials.The comparison of different methods of analysis for the same samples is increasing however. The results obtained for ETA/AAS were compared with those from NAA for the analysis of A1 in urine and bone (642), and for Au in plasma and plasma fractions from rheumatoid patients receiving chrysotherapy (299). If standard samples were incorporated into such studies, CRMs with a high degree of authenticity could be produced.Considerable losses o f analyte can occur both during sample preparation and in the ashing stages of analysis using ETA. It is essential to minimize these losses, which some- times occur because the sample material is subjected to high temperatures, which in different circumstances have been found not to cause problems. However, errors can also occur through the use of poorly calibrated muffle furnaces for ashing the sample.Similarly, the temperature attained during pre-atomization stages in ETA i s often very different from that indicated by the instrument, or obtained from a calibration graph. Recent advances in instrumentation (240) have helped to overcome some of these problems.Smith et al. (101) have pointed out that the percentage of total Au can be very variable in different serum protein fractions, and that it would appear that the measurement of serum Au level does not adequately reflect the amounts of the various Au forms present in serum. The Au was determined in this study by ETAIAAS. There is increasing interest in autosQmpling devices for both flame and flameless AAS.Autosampling would appear to offer potential benefits in both speed and precision. Several workers have described automatic equipment or its applications (319, 798; 1282). Two points would appear worthy of amplification. First, there is little gain in throughput with auto- sampling into a flame. A major advantage of such systems is said to be the saving of operator time but, especially when different dilutions are required for different elements, the percentage of time saved on the total analysis is minimal.However for ETA at relatively low temperatures, where the furnace might be run without close supervision, autosampling systems running outside working hours could significantly increase the sample throughput. Once again limitations occur if, for example, dilution or changes in element are required.Secondly, autosampling systems may also lead to an improvement in pre- cision. For flame systems, however, where the precision of manual analysis is generally adequate, such improvement may be of little practical significance. For ETA, autosamplers do appear to make a useful improvement in the precision obtainable in routine analysis. 4.8.2 Sample Preparation The analysis of body tissues and fluids poses many problems for the analyst and so it is not surprising to see a wide range of sample preparation procedures reported in the literature. Despite increasing awareness of the hazards involved in the routine use of perchloric acid, its effectiveness in breaking down biological materials means that it is still widely used, Julsham (698) studied the effect of HClO, on the determination of 10 commonChapter 4: Applications 165 elements by ETA.Signal suppression ranged from 8% for Zn to 100% for Al. For all the elements studied the effect could be overcome by the addition of 5% of €€NO,. Solvent extraction continues to be a useful tool. Beryllium has been determined by ETA after extraction as the acetylacetonate into MIBK in the presence of EDTA.The detection limit was ca. 1 ngml-1 (548). Cadmium, Cr and Pb have been extracted with APDC into MIBK followed by back extraction into 10% KNO, prior to analysis (1169). The instability of APDC complexes has not previously been adequately stressed and workers in other fields (see 4.7.2) have found back extraction to be imperative in some instances. Examples of the importance of attention to the influence of sample preparation equipment have again been reported.Urquhart (1 156) observed considerable contamination of his control serum from Zn in rubber stoppers. Healy et 01. (1167) found that silicone- treated stoppers interfered in the determination of serum Cu by delaying the release of Ch from protein.Smith (1 3%) reported severe contamination from borosilicate glass in the determination of Cd in N.B.S. orchard leaves. Lyophilization has been used in the determination of Hg in marine biological samples (571). A method has been reported for the determination of Zn in cerebrospinal fluid by ETA, following ion-exchange chromatographic separation (627’). 4.8.3 Atomic Absorption Spectroscopy The need for accurate Correction for non-specific absorption has steadily increased in importance and it is now rare to see an instrument for AAS with no facility for background correction using either a deuterium HCL or arc lamp. Although in many ways this has led to improved analyses, the associated problems of accuracy are still being discussed.Zeeman-effect background correction has been suggested as a more accurate method; the merits of this technique are also still open to some debate, although it is now being used for routine analysis in some laboratories (930). Whenever analyte concentrations are suitable, FAAS methods are generally preferred owing to their speed and precision, Continuing advances in instrument design have meant that some analyses formerly requiring ETA can now be performed using flame atomization.The determination of Ag in blood using the Delves cup technique has been described (712). The detection limit was 2.7 ng ml-1 and the between batch RSD was 0.08 at 10-50 ng ml-1. There has been considerable interest in the determination of As. Chrenekova and Rusinova (1031) used a AgDDC extraction of As into CHCI,, followed by evaporation and dissolu- tion in 4.8M HCl, before determination of the element in an air/C,H, flame.Several variations of hydride generation techniques have also been described for As determination (740, 369, 12543, whereas Ishikazi and Kataoka (591) used ETA, Mushak et al. (3009 investigated the determination of monomethyl and dimethyl As, and inorganic As in different oxidation states.The organic As compounds were determined by GC-ETA. The Delves cup technique has been of great importance for the determination of Pb in blood; however for several reasons it required extremely experienced operators to produce results of high precision. Thus as ETA methods have improved there has been considerable interest in the possibility of the direct determination of Pb in blood by this technique.Some papers have appeared during 1978 which described such methods, the most notable being the work by Stoeppler et aZ. (532) in which they achieved an RSD of 0.08 at 1OOpg1-1 and 0.03 at 480 pg 1-1. It is possible to determine copper in biological fluids using FAAS or ETA (81) and in general the simpler flame method is to be preferred.However, when low levels are present, ETA is essential and the RSD of 0.04 (at 50 ng ml-1) obtained by Kame1 et al. (71 3 ) shows what can be achieved under ideal circumstances. Villet et 01. (1 110) used Cu in the indirect ~166 A n aly tical A tom ic Spec trosco p y determination of antihistamines by atomic absorption, The antihistamines were complexed with a known amount of Na dioctylsulphosuccinate and the excess of detergent was com- plexed with Cu 1,lO-phenathroline.The latter complex was extracted into MIBK and the Cu content determined. Campe et ul. (966) described a rapid (less than 6min) method for the determination of Hg in various forms and total Hg in the same portion of urine by cold-vapour AAS.Inorganic Hg was determined in 15% NaOH containing 0.03% L(+)-cystine; inorganic and phenyl Hg were determined in 32% H,SO,. Total mercury was determined after digestion with KMnO, and H,SO,; an RSD of 0.01 was claimed. An improved method for the determination of Co in biological samples using ETA has been reported (1215). Ashed samples were dissolved in 10M HC1, extracted with a CCl, solution containing 5% tri-n-octylamine, and then re-extracted with H,O.The sensitivity for Co was 0.1 ppb and there was no significant interference from As, Cd, Fe, Ni, Pb, Sb, Se or Zn. The RSD was 0.10 and the recovery >W%. A method has been proposed for determining Ga in serum and tissues from humans and animals receiving Ga for the treat- ment of cancers (1 159).The recovery was better than 95% for 5-50 p g Ga added to 0.5 g tissue. Although 0.5mM Ca suppressed the signal from 5ng Ga by SO%, this effect could be overcome by the addition of EDTA. Bonilla (1 152) determined Mn in rat tissues using ETA, The sensitivity was 1.16 X 10-11 g and the RSD was 0.047 for 0.5 ng. Adams et al. (1153) produced a valuable paper on interlaboratory comparisons of Ni in urine by ETA-AAS.Those procedures involving preliminary oxidation and extraction stages were generally superior to direct ETA methods on the basis of recovery of added Ni, interlaboratory precision and sensitivity. Hurlbert (646) determined Be in a range of biological materials using ETA and found it necessary to add La at 100pgm1-1 to minimize the effects of a number of interferents.Cr was determined in the urine of ‘unexposed’ subjects by ETA (47’8); the mean level found was 2.2ppb. The use of cis-dichlorodiamine Pt as an anti-cancer drug has encouraged two papers on the determination of Pt in serum (215, 1154), in both of which ETA-AAS was used. The sensitivity of V using ETA was found to be very dependent on the chelating agent used in its extraction (121 3, 1214).The best results were obtained with n-benzoyl-o-tolylhydroxl- amine. Workers interested in the determination of Zn in serum should find two recent papers of great interest (1151, 1161). Both compared a direct ETA method with one using TCA for protein precipitation. Both obtained quantitative recoveries of Zn using dilution and ETA and high results using TCA; different explanations were advanced for the high TCA results in the two papers. The year has seen the appearance of a new technique metallo-immunmsay (7). The disadvantages of radioisotopes have stimulated the search for non-isotopic methods in immunological research.The authors believe they have found a practical alternative to radioimmunoassay utilizing AAS. Specially prepared metallohaptens react with antigens and antibodies, and the metal content of the resulting complex is monitored spectro- scopically.It will be interesting to see how the technique develops. 4.8.4 Other Methods Atomic fluorescence has had a chequered history but it has now emerged as a technique which can complement AAS and provide superior results in some instances.Lipan and Plankey (149, 946) used this method to determine Cu and Mg in blood serum and workers at Strathclyde have produced several papers on the applicability of AF to clinical samples (214, 716).Tabls 4.8 BODY TISSUES AND FLUIDS form Element X/nm Matrix Concentration Tech. Analyte Sample treatment Atomization Ref. Ag Al Al A1 Al Al As As As As As As 328.1 Blood - Serum 309.3 Bone, urine 309.3 Biological fluids and - Biological fluids tissues - Tissue - Body fluids and tissues - Soft tissues 193.7 Urine - Blood - Fish tissue 193.7 Biological material As - Animal tissue 3-400 ng/ml Up to 400 bg/I From 0.1 pg/g - - 0.5 ng/ml and above 1.2-74 pg/g 530 , d m l (in extract) A A A A A A A A A A A A A L L L L L L G L G L G L None F Air/C,H, 712 (Delves cup) Graphite furnace 290 Ur i ne-d irec t met hod Graphite furnace 642 Bonedigest at 60 "C with HNO, f CaCI,/K,HPO, solution (HGA-76) (HGA-2100) Graphite furnace 1120 Graphite furnace 1140 F N,0/C,H2 1248 Graphite furnace (CRA-90) (HGA-74) 279 Hydride-evolution method 4 - Study of speciation analysis for Graphite furnace 300 inorganic, m onomet hy I- and dime: hy I- arsenic.Results compared with those from GLC studies Treat with HCI + NaBH,; pass evolved F Air/H,/N, 369 ASH, through Pb acetate solution to remove H,S Dry, digest with HCI and extract As Graphite furnace 591 Into CHCI, layer.Redissolve in Mg( NO,), solution Ash (LTA), prepare solution i n Heated tube 740 2M ECI and proceed with ASH:- evolution method Ash with Mg(NO,), + NH,NO, at F Air/C,H, 1031 500 "C.Redissolve. extract with AgDDC/CHCI,, evaporate to dryness and dissolve in HCI L, G Digest with HNOJHCIO,. Determine F - 1105 As by hydride-evolution method and Cd, Pb by FAASc m 00 Table 4.8 BODY TISSUES AND FLUIDS-continued Element X/nm Matrix Concentration Tech. luaruleail eldues Atomisation Ref. form As As A4 Au Au A u. B e Be Ca Ca Ca Ca Ca Ca Ca Ca - - - 242.8 242.8 242.8 234.9 - - - 422.7 - - 422.7 622 422.7 Body fluids, tissues From 10 ng/ml Fish tissue - Blood serum, white cells 0.01-3 mg/l Biological f I u id s - Plasma - Serum 74-1036 hg/lOO ml Biological materials ng/g levels Biological materials ng levels (absolute) Blood Blood serum Biological samples Urinary calculi Blood Serum Serum Serum - 2.2-3.4 mmol/l - 4.7-5.5 mmol/l 4.7-5.5 mmol/l A A A A A A A A E A A A A A E A G G L L L L L L L L L L L L L L Digest with acid and add NaBH, Homogenize, treat with H,O,, followed by H,SO,/KMnO, and &S208- Clear with NH,OH.H,SO,/NaCI and reduce with NaBH, to generate ASH, vapour Heat with saturated KMnO, solution 4- 6M HCI and extrrct with MIBK Comparison of NAA and ETA-AAS Dilute (1:3) with 0.1N HCI Ash at 500 "C, dissolve in HCI and extract Be at pH 6 as acetylacetonate with MIBK, in presence of EDTA.Back-extract with 1M HCI (A) Tissues-digest with HNOJHCIO, - and evaporaie to dryness. Redissolve in HNO,/H,O (B) Faeces-digest with Fe(ll) ion (C) Urine-Co-precipitation method Add La (100 pg/ml) to all solutions Automated system for Ca, Na, K Add LaCI, solution (Automated system) Heat, on water bath, with 1M NaOH and add La + EDTA See Cd, ref. 445 Add KCI to final concentration of 2000 pg/ml Aulomated sys:em, incorporating marker ion (Na) internal standard circuit Study of ascorbic acid interference Add LaCI, solution + H A Heated SiO, cell Heated SiO, cell Graphite furnace Graphite furnace GrapHite furnace F Air/C,H, Graphite furnace Graphite furnace (HGA-210) F - F - F Air/C,H, F - F N,O/'C,H, F Air/C,H, F - F Air/C,H, 1254 1401 101 197 299 598 548 646 309 $ 339 5 352 3 EL 445 b 470 $ E * 798 c/, 2 2 799 aoo 2Ca 239.9 Pancreas Ca 422.7 Serum Cd - Blood; urine Cd - Blood Cd Cd Cd Cd Cd cd Cd Cd Cd - Blood, urine - Blood, urine - Blood - Urinary calculi 228.8 Blood 228.8 Blood 228.8 Bone 228.8 Serum, urine - Blood, urine Cd - Serum - A S - A L 5 pg/l level A, F L - A 5 pg/l level c F F 0.45 &g/g level A A F - A L L L L L L L L L L L Incubate tissue in bicarbonate buffer (pH 7.4), wash with saline and freeze- dry.Take 2-20 pg solid samples Inter-laboratory study Dilution only Ramp-dry to 285 "C and atomize at 1500 "C (Example of application of precise temperature control system for furnace) - Blood-Haemolyse and dilute 1.4 with 0.04M HCI containing 0.5% Triton-X Urine-Nebulize directly Dilute (1:l) with acid and heat at 75 "C for 10 min.Powder and digest with HCI and/or HNO, Add HNO,. For flame method, chelate and solvent-extract Add Triton X-100, transfer to cup. dry at 120 "C and heat to 400 "C before analysis Treat with ethyl ether, dry at 110 "C under reduced pressure, dissolve portion in HNO,, add H,O, and Graphite furnace F - F - Graphite furnace F Air/C,H, F Air/C,H, (argon- separated) Graphite furnace F - Graphite furnace P - F Delves cup F Air/C,H, evaporate to dryness.Ashat 350-400 "C, dissolve in acid and extract with APDC/MIBK at pH 2-3 (Cd. Cu, Zn) - Graphite furnace (A) Blood-Haemolyse and dilute F Air/C,H, (argon- (HGA-2000) (1:4) with 0.02M HCI containing 0.5% Triton X separated) (5) Urine-Aspirate directly Dilute (1:l) with H,O.Prepare F Air/C,H, standards in H,O + 0.5% NaCl 4- 20% glycerol 984 1306 21 4 240 290 5 31 9 445 455 500 550 630 71 6 831Tablz 4.8 BODY TISSUES AND FLUIDS-continued c 0" Element X/nm Matrix concentration Tech. Anaiyte Sample treatment Atomisation Ref. form Cd Cd Cd Cd Cd Cd Cd Cd Cd c o Cr Cr Cr Cr c u c u c u c u c u 228.8 - - 228.8 228.8 - - - 228.8 - - - 357.9 357.9 I - I 324.7 - Bone Kidney tissue Animal tissue Fish tissue Blood serum Blood Urine Biological samples Lung tissue Biological materials Urine Serum Fish tissue Blood serum Fish tissue Blood serum Biological materials Bone Fish tissue, waters A - From 1 ng/g A - A 0.1-1 A From 5 ng/g A - A 1-100 nmol/l A Trace levels A - A From 0.1 ng/g 2 ng/ml level 1.34 pg/ml 1 &g/g level - A A A A A A F A A A L L L L L L L L L L L L L L L L L L S, L Digest with HNO, in PTFE bomb Graphite furnace 892 Digest wi:h HNO, (Zeeman-effect ETA system) Dry-ash at 500 "C, dissolve and add pH 2.4 4 0.2 and extract with APDC/MIBK.Back-extract with Dilute (1:l) with H,O F Air/C,H, Interference study Graphite furnace Dilute (1:l) with 0.3M HNO, Graphite furnace Contamination study Graphite furnace Freeze-dry, digest with HNO, in PTFE F Alr/C,H, bomb, partially neutralize with NH,OH and dilute.Calibrate by standard addition method Ash, dissolve in HCI, extract with Graphite furnace TOPO/CHCI, and back-extract with H,O Add HNO, to overcome interference Graphite furnace by NaCl See Cd, ref. 831 F Air/C,H, See Cd, ref. 1169 Graphite furnace See Cd, ref. 1204 F Air/C,H, Digest with HNO, at 150 "C in pressure F Air/C,H, vessel. Aspirate using pulse-nebulization technique Application of oscillating filter atomic F - fluorescence spectrometer Application of WC-coated crucible Graphite furnace See Cd, ref. 550 F Air/C,H, Graphite furnace See As, ref. 1105 F - Graphite furnace KMnO, t o form Cr(VI).Adjust to (CRA-63) 10% HNO, (CRA-63) - - - 930 1105 1169 1204 1212 1262 1386 1 388 1215 478 b 831 5 1169 1204 3 b 1 1 %. 9 149 6' c/, 516 % Y 550 2 626 .$ vc u cu c u cu c u cu cu cu cu Fe Fe Fe Fe Fe Fe Ga Ga 324.7 Serum, urine 324.7 Faeces 324.7 Plasma - Kidney tissue 324.7 Blood serum - Plasma - Serum - Rat liver - Biological samples 248.3 Metallohapten solutions - Fish tissue - Urinary calculi - Rat tissue 248.3 Urine 248.3 Serum 294.4 Biological fluids and 417.2 Urine, serum tissues tissues 20-120 ng/ml (20 hI sample) - From 10 ng/g Up to 7.5 pg/ml pg/m I levels 5-12 fimol/l Trace levels &50 ng/ml (in extract) 0.7-151 pg/g - From 42 ng (absolute) 20-60 ng/ml (20 f i l sample) 14.3 fimol/l 100-200 pg/l A A A A F A A A A A A A A A A A A L L L L L L L L L L L L L L L L L Homogenize (1:l) with H,O, freeze, dry at 60' C, ash (LTA) and dissolve in 1N HCI. Dilute x10 for analysis.Results compared with wet-ashing (HNO,/HCIO,/H,SO,) method Method for ultrafiltrate samples See Cd, ref. 930 Wavelength-modulated continuum source AFS system Study of Cu/Zn ratio (A) Wet-ash with HN03/HCI0,/H2S0, (6) Precipitate proteins with TCA and analyse supernatant liquid (C) As (B), but centrifuge and continue as in (A) See Cd, ref. 1386 Applicarion of flameless AAS to metalommunoassay studies See Cu, ref. 11 See Cd, ref. 445 Heat with HNO, in sealed PTFE vessel and dilute with H,O Acidify to pH 2, dry at 60 "C and ash (LTA). Dissolve in 1N HCI (See also Cu, ref. 696) Deproteinize (Comparison with spectrophotometric method) Digest with HNO,, filter and dilute (tissues) or, dilute with H,O and add EDTA (fluids) Graphite furnace Graphite furnace (HGA-2000) (HGA-2000) Graphite furnace Graphite furnace F Ar/air/C,H, (HGA-72) F Air/C2H, F - F - Graphite furnace Graphite furnace F Air/C,H, Graphite furnace (HGA-70) F - Graphite furnace (HGA-2000) F - Graphite furnace Graphite furnace (CRA-90) ( FLA-10) 630 696 71 3 930 946 1098 1162 1259 1386 7 11 445 490 696 830 1120 1159+ Table 4.8 BODY TISSUES AND FLUlDt3-continued -4 p3 Element X/nm Matrix Concentration Tech.Sample treatment Atomization Ref. form - Biological samples - Biological fluids - 7 pg/l level A A G L - Cold vap-our Method for total, inorganic and organic - - mercury 290 306 - Urine - Biological samples 253.7 Fish tissue - Fish tissue 462 467 571 702 - 0.02-0.65 I g / g 0.15-500 ng/g - G G - - - Digest with HNO, in pressure vessel Wet-digest with KMnO,/H,SO,, add NH,OH.HCI and reduce with SnCI, in Digest with dilute NaOH solution Cold vapour and reduce with SnCI, Flameless method for alkyl mercury - - Digest with KMn0,/H2S04, add Cold vapour NH,OH.HCI and reduce with SnCI, Pyrolyse in stream of N, and pass Cold vapour vapour products t o combustion zone to burn in stream of 0,.Pass vapour products through N%CO,, collect Hg by amalgamation with Ag and release by reheating Cold vapour Cold vapour H F 4 Applications of Magos method - - Collect on Ag wool and release by rapid Cold vapour graphite cuvette heating Mix with inert material, e.g., sand, Cold vapour and heat in 0,.Pass vapours through acid KMnO, - Cold vapour See Ca, ref. 309 F - Automated simultaneous method F - for Na, K See Cu, ref. 149 F - See Ca, ref. 339 F - - Blood 0.03 ng/ml A G 71 9 - Fish tissue 253.7 Urine A A 860 966 - G - Biological materials A G 983 - Biological materials - Biological materials A A 1222 5, - From 0.01 ng/ml - G 1247 & 1398 ‘1, -.. - Biological materials A G L - 1401 309 7 - Fish tissue - Blood - Blood, serum, urine Hg K K A E E L, G L L - Blood serum - Blood serum 25.1 pg/ml 1 .O-1.3 mmol/l F A L LMn 279.5 Mn - Na - Na - Ni - Ni - Ni - Pb - Pb - Pb - Pb - Pb - Pb - Biological samples Urinary calculi Blood, urine Blood Serum Blood serum Urine Blood Bone Kidney tissue Biolog ica I mat er ia I Animal tissue Rat liver Blood Blood, serum, urine Urine Urine Plasma, urine Tissues (lung, liver, kidney) Blood Blood Red blood cells Blood Blood pg/ml levels 100 pg/l levels - - - - 2 pg/l level A A A A A F A A A A A A A E E A A A A A A A A A L L L L L L L L s, L L L L L L L L L L S L L L L L See Ca, ref. 352 See Cd, ref. 445 Add LaCI, solution See Ca, ref. 470 See Ca, ref. 798 See Cu, ref. 946 Chelation-extraction method See Cd, ref. 892 See Cd, ref. 930 Wei-ash with HSIO, Digest with HNO, See Ca, ref. 369 See K, ref. 832 Wet-digest and extract Ni by chelation- solvant extraction Results of inter-laboratory study; oxidation and extraction methods preferred to direct methods Precipitate proteins with TCA + H,SO, and extract Ni with APDC/MIBK Homogenize with water.Prepare standards (up to 0.65 pg/ml) i n blood Review. See Al, Cd, Hg, ref. 290 See Cd, ref. 319 Comparison of methods F Air/C,H, F - F - F N,0/’C2H, F Air/C,H, F Ar/air/C,H, Graphite furnace Graphite furnace Graphite furnace Graphite furnace Graphite furnace Graphite furnace (HGA-72) (HGA-2100) F - F - F - Graphite furnace 352 445 458 ’z 470 ‘ 798 2 577 3. 753 2 892 946 930 1 G97 1152 1259 309 832 463 Graphite furnace 1153 Graphite furnace 1233 F Air/C,H, 233 + Delves cup F - 280 F - 290 Graphite furnace F - 298 Graphite furnace Graphite furnace 319 F (Delves cup) 407 Graphite furnace c 4 wTable 4.8 BODY TTSSUES AND FLUIDS-contiizrred P EIement X/nm Matrix Concentration Tech* Anaiyte Sample treatment Atomisation Ret. form Pb P b P b P b P b P b P b P b P b P b P b P b P b P b P b Pb - Blood - - Urine Ng/I levels 283.3 Blood 100-500 pg/l 217.0 Serum, urine 283.3 - Blood - Blood - Bone, ivory - Blood 30 &I00 m l (average) - Kidney tissue - 283.3 Blood 0-120 f i g / % - Blood - - Blood 0.03-0.52 pg/lOO ml - Animal tissue - 217.0 Blood 0.42-2.99 kmol/l - Teeth; water 8upplies - 283.3 Tissues 0.035-1 ng/g A E A A A A A A A A A A A A A A L S L L L L s, L L L L L L L L L L See Cd, ref 455 Add TI solution (internal standard), freeze-dry and excite in graphite crucible by double-arc method Add Na citrate.Dilute sample with H,O + HNO, (2:3:3), shake, centrifuge and analyse Comparison with ASV method Treat 2 m l sample with TCA/HCIU,, centrifuge and add HNO, lo supernatant liquid before analysis See Cd, ref. 832 Comparison of 4 treatments See Cd, ref. 930 Mix ( 1 4 ) with Triton X-100 Mix with NH,NO, or “lumatorn” before ETA analysis See As, ref. 1105 Dilute heparinized blood (1:4) with 1% Triton X-100 solution, add HNO,, stand for 30 min. and take 20 PI aliquots for analysis I Mix (1:4) with 50% HNO,, stand overnight, adjust pH with NaOH + NaHCO, (to slight excess), add KCI and extract P b with APDC/MIBK F - Graphite furnace A - Graphiie furnace (HGA-74) Graphite furnace Graphite furnace (HGA-2000) F - Graphite furnace F Air/C,H, Graphite furnace Graphite furnace Graphite furnace Graphite furnace Graphite furnace F - Graphite furnace (CRA-63) Graphite furnace (HGA-72) F Air/C,H, 455 492 532 630 762 856 892 900 930 970 1041 1101 1105 5.b % 5 5 1158 5 1160 ‘* t.l 1168 !$ 2 2 QPb Pb Pb Pb Pb Pb Pt Pt Sb Sb Sb Se Se Se Se Se 283.3 - - - - 227.0 265.9 265.9 - 217.6 - - I - 196.0 - Fish tissue Blood Blood Blood Blood Lung tissue Biological materials Plasma Blood Blood, urine Biological materials Biological selenoids Body fluids and tissues Biological materials Biological materials Body fluids, tissues From 0.02 ,ug/ml - From 30 ng/g - 0.5-5 ng/ml From 0.3 ng/g - 2 ng (absolute) 0.05-1 b9/9 A A A A A A A A E A A A A A A A L L L L L L L L L G L L G G L G See Cd, ref. 1169 None Add quaternized NH,OH or NH,NO, Comparison of methods See Cd, ref. 1388 Prepare ultra-filtrate, add EDTA, separate on cation-exchange resin paper and elute with 5M HCI Dry sample (1.0-1.5 ml), ash (LTA) and dissolve in HCI. Chelate-extract into CCI, and back-extract with H,O containing trace Cu.Transfer aliquot to SiO, tube, freeze and seal in tube in inert gas at 2 torr pressure SbH, generation method Ash, extract with xylene containing 5% Amberlite LA-1 and back-extract with 0.1M tartaric acid Combined LC/Zeeman-effect AAS detector system, used for speciation of Se compounds See As, ref. 279 Hydride method Combust in 0,-flask containing 20 m l 0.01M HCI.Separate Se on IR-120 cation-exchange resin column, reduce with NH,OH.HCI and extract as Se( IV) with dithizone/CCl,. Add Ni solution to prevent loss of Se at ashing stage of ETA analysis See As. ref. 1254 Graphite furnace 1169 % Ta ribbon 1245 $ Graphite furnace 1263 ? F Delves cup 1267 b Graphite furnace Graphite furnace 1316 ? Graphite furnace 1388 -- Graphite furnace 215 Graphite furnace 1154 (CRA-63) e % 2 P (CRA-90) P MIP 741 (2450 MHt) F Air/C,H, 1077 + SiO, tube Graphite furnace 1216 F - 55 - - 279 Heated SiO, tube 720 Graphite furnace 1145 Heated SiO, cell 1254 cTable 4.8 BODY TISSUES AND FLUIDS-continued Element X/nm Matrix Concentration Tech.Anal’te Sample treatment Atomization Ref. form Sn V Zn Zn Zn Zn Zn Zn Zn Zn Zn Zn Zn Zn Zn Zn Zn Zn - Biological samples - Biological materials - Fish tissue - Blood, plasma, erythrocytes - Blood, urine - Plasma 213.9 Bovine liver 213.8 Bone 213.9 Cerebrospinal fluid - Serum - Saliva, blood, plasma 213.9 Serum 213.9 Serum - Serum, plasma, urine 213.8 Serum 213.9 Blood, serum - Rat liver Plasma From 0.05 ng (absolute) 7.4-24.9 pg/g - 350 pg/g level Trace levels - pg/ml levels 100-1500 p g / l 1-4 mg/l 0.4-15.0 pmol/l 40-200 p g / l From 3 ng/g 900 ng/ml level - A A A A F A E A A A A A A A A A A A L L L L L L L L L L L L L L L L L L Method for bis-(tri-n-bu:yl tin) oxide Graphite furnace Study of chelation-extraction methods Graphite furnace See Cu, ref. 11 F Air/C,H, Dilute blood of saline-washed F Air/C,H, erythrocytes (1:19) with 0.05% Triton X-100 solution.Dilute plasma samples (1:4) with H,O See Cd, ref. 5 F - - F - Ash (LTA) and dissolve in HNO, See Cd, ref. 550 Digest i n PTFE dish with HCIO,, fume, add HCl/methanol and pass through Dowex 1-X8 column. Wash w’ith 1M HCI and elute Zn with 0.5M HNO, See Cd, ref. 831 Dilute (1:lO) with H,O, or treat with TCA and analyse supernatant liquid Dilute (1:5) with H,O (serum, plasma) or as necessary (urine) Mix ( 2 : l ) with 0.6M TCA, heat to 90 “C and analyse supernatant liquid (flame method) or dilute (1:lO) with H,O (ETA-method) See Cd, ref. 1204 P Microwave plasma F Air/C,H, Graphite furnace (CRA-70) F Air/C,H, F Air/C,H, F Air/C,H, F Air/C,H, F Air/C,H, F Air/C,H, Graphite furnace (CRA-63) F Air/C,H, F - Contam inat ion study F - 655 1213 1214 1 1 303 5 460 549 550 627 831 1098 1151 1156 1157 1161 1204 1259 1264Various - Biological samples Trace levels (5) (blood, urin,e, liver tissue) Various Various Various Various ( 8 ) Various Various (233) Various - Clinical and biological Trace levels samples - Biological samples -- - Clinical samples Trace lsvels - Serum - - Biological materials - - Clinical samples - - Heparin; insulin - Various - Faeces (8) Various - Fresh water organisms Trace levels Various - Biological fluids Trace levels (12) Various - Biological materials - ( 1 2 ) Various - Brain tissue Trace levels Various - Fish tissue (6) Trace levels Various - Biological materials - E .4 A F A A A A, E A A A A, E E A E G L L L L L L L L L L L S L L Combined system, involving hydride P Microwave generation, chroma!ographic separation plasma (Chromasorb 102, 108) and microwave plasma excitation (in Ar/He flow).Elements: Ge, As, Se, Sn, Sb Review; 15 refs. F - Graphite furnace Study of sample treatments in relation Graphite furnace t o “charring” stage of atomization cycle Discussion of design and application of F Air/C2H, a flame atomic fluorescence spectrometer (Ar-separated) - F - Comparison of methods F - B!ood, plasma, urine, etc.-dilution F - only (Review paper) Graphite furnace Various determinations, including Na, K F - in heparin from lung tissue; Fe, Cu in insulin from pancreas tissue and in ACTH from pituitary tissue (A) Digest with HCI/HNO,/H,O (27:3:20) ( 5 ) D.gest with HNO,/HLO (1:20) (C) Dry-ash at 450 “C and dissolve in HCI F Air/C,H, N20/’S,H, Ash and re-dissolve F - Various (direct injection, ashing and extraction-concentration ) Method for Be, Cd, Cu, Mg, Mn, Hg, F - Ni.Zn by AAS and Ca, K, Rb, Na by FES - Laser microprobe Graphite furnace - Review I - - P ICP 56 81 108 144 278 281 80 324 31 0 409 41 3 507 574 61 7 625 662Table 4.8 BODY TISSUES AND FLUIDS-continued c 2 Element X/nm Matrix Concentration Tech.Sample treatment Aiomisation Ref. form Various (10) Various Various Various Various Various Various Vnrious Various Various (5) (8) Various Various Various (7) Various (3) Various Various ( 4 ) Biological materials - Biological materials - Biological materials Trace levels Biological materials Major and trace Marine organisms Trace levels levels Seric proteins p g / g levels Plasma Trace levels Fish plasma - Blood platelels - Serum - B i o I og ic al mat er i a Is Biological fluids - - Coral tissue and skeleton - Biological materials - (serum) Blood.urine Trace levels Biological materials Trace levels A E A A A A A E A A A A A E E A L L G L L L L L L L L L L L L L Study of effect of HCIO, Review, with references to Pb, Cd, Cu, Mn i n human placenta; Pb, Cd i n hair; Li, Cr, Mn, Zn i n blood, serum and TI in urine Review of hydride-evolution method Review Digest with HNO, i n PTFE bomb Sapara:e proieins by elec:ro>hcrssis and extract fractions with HNO, - Add EDTA.Results for Fe, Cu, Zn, Ca, Mg Pulse-nebulization method (50 or 100 p! samples) for Na, K, Mg, Ca, Li, Cu, Zn, Fe Comparison of AAS, XRF and NAA Treat with H,O, NaOH, NH+OH t o remove organic material from skeleton. Results given for Cd. Co, Fe, Mn, Ni, Pb, Zn D ~ l u t e (1:4) with 0.01M HCI containing Mn as internal standard Concentrate impurities with a po y(di.h,oc.rbamate) resln treatment Mineralize with acid and extract (Methods for Cd, Pb, Sn, Zn) Graphite furnace 2 ICP (HGA-76) Heated tube Graphite furnace Graphite furnace Graphite furnace F - F - F - F - F - F - Graphite furnace Graphite furnace F - P ICP P ICP Graphite furnace (HGA-72) 698 724 742 835 836 1052 1155 608 1246 12112 1287 1304 1308 '1, s, ;i' 2 1419 ? 3 5' 1420 tl 1471 5 B 8 *aChapter 4: Applications 1 79 A microwave induced plasma has been used to measure Sb in blood (7'41). Arsenic, Ge, Sb, Se and Sn have been determined in various biological samples, after conversion to the hydrides and chromatographic separation, using an Ar/ He MIP (56). Lmer-excited emission spectrography has been employed to study the levels of various metals in sections of human brain (6173, the results being described as comparative rather than semiquantita- tive. Cecchetti et d. (942) used d.c. arc emission spectrography to determine Pb in urine, while Nakasuma (549) determined Zn in bovine liver with a CMP torch. Several workers (662, 724, 1419, 1420, 1428) have utilized the increasingly popular ICP for the analysis of biological materials. There is growing evidence to suggest that chromatography may have a role to play in atomic spectroscopy, both when matrixes are difficult and when the speciation of the element is of special interest. Vickey and Howell (55) used a Zeeman-effect atomic absorp- tion spectrometer as a specific detector following the LC separation of biological Se compounds.