# Elements in Blood or Tissue (8005)

​ELEMENTS in blood or tissue
M.W.: Table 1
8005
FORMULA: Table 1 CAS: Table 1
RTECS: Table 1
EVALUATION: PARTIAL
Issue 1: 15 May 1985 Issue 2: 15 August 1994
METHOD: 8005, Issue 2
BIOLOGICAL INDICTOR OF: exposure to the following elements or their compounds: antimony, cadmium, chromium, cobalt, copper, iron, lanthanum, lead, lithium, magnesium, manganese, molybdenum, nickel, platinum, silver, strontium, thallium, vanadium, zinc and zirconium.
SYNONYMS: vary according to compound.
BIOLOGICAL SAMPLING SPECIMEN:
BLOOD OR TISSUE
VOLUME:
10 mL (blood) or 1 g (tissue)
MEASUREMENT METHOD:
INDUCTIVELY-COUPLED ARGON PLASMAATOMIC EMISSION SPECTROSCOPY (ICP-AES)
PRESERVATIVE: heparin (blood); none for tissue
ANALYTE:
elements above
SHIPMENT:
frozen for blood and "wet" tissue; routine for "dry" tissue
DIGESTION ACID:
STABILITY:
not established
CONTROLS:
collect at least 3 blood specimens from unexposed workers
3:1:1 (v/v/v) HNO 3:HClO 4:H2SO 4
FINAL SOLUTION:
10% H 2SO 4; 10 mL (blood), 5 mL (tissue)
WAVELENGTH:
varies with element; Table 2
BACKGROUND CORRECTION: spectral wavelength shift CALIBRATION:
elements in 10% H 2SO 4 or yttrium internal standard
QUALITY CONTROL:
spiked blood or tissue; reference materials
RANGE:
10 to 10,000 µg/100 g blood; 2 to 2000 µg/g tissue
ESTIMATED LOD:
1 µg/100 g blood; 0.2 µg/g tissue
PRECISION (Sr):
Table 2
ACCURACY:
Table 2
APPLICABILITY: This method is useful for monitoring the blood of workers exposed to several metals simultaneously. This is a simultaneous multielemental analysis, but is not compound-specific.
INTERFERENCES: Spectral interferences are sometimes encountered. These are minimized by judicious wavelength selection and interelement correction factors. Background corrections (spectral wavelength shift) are also made [1,2].
OTHER METHODS: This method uses a measurement technique similar to that of Methods 7300 (Elements; for air samples) and 8310 (Metals in urine).
NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94 ​ELEMENTS in blood or tissue: METHOD 8005, Issue 2, dated 15 August 1994 - Page 2 of 6 REAGENTS: 1. 2. 3. 4.
5.
6.
7. 8. 9.
EQUIPMENT:
Nitric acid, conc. (HNO 3) (high purity).* Perchloric acid, conc. (HClO 4) (high purity).* Sulfuric acid, conc. (H 2SO 4) (high purity).* Digestion acid, 3:1:1 (v/v/v) HNO 3:HClO 4:H 2SO 4.* Mix three volumes conc. HNO 3 with one volume conc. HClO 4 and one volume conc. H 2SO 4. Element standards, 1000 µg/mL. Commercially available or prepared per instrument manufacturer's recommendations. Reference materials of known elemental composition. SRM #1577a, Bovine Liver, from the U.S. National Institute of Standards and Technology is recommended. Argon. Deionized water. Yttrium standard, 5 µg/mL in 5% HNO 3. Combine 50 mL conc. HNO 3, ca. 500 mL deionized water, and 5 mL 1000 µg Y/mL standard. Dilute to 1 L.
See SPECIAL PRECAUTIONS.
1. Blood collection tubes, heparinized, lead-free, specially prepared for collecting blood samples for blood lead determinations. NOTE: Heparin may contain significant amounts of Ca, Cu, Mn, Sr and Zn and should not be used if these elements are to be determined [3]. 2. Vacutainer needles (21-gauge) and holder. NOTE: These may be significant sources of contamination in some cases [4]. 3. Tourniquet and alcohol swabs. 4. Bottles, glass or polyethylene, with PTFE-lined caps, 20-mL (e.g., scintillation vials).* 5. Inductively-coupled plasma-atomic emission spectrometer equipped for determination of elements of interest. 6. Regulator, two-stage, for argon. 7. Analytical balance, readable to 1 mg. 8. Beaker, Griffin (50-mL) or Phillips (125-mL), with watchglass covers.* 9. Hotplate, for use at 110 and 250 C. 10. Pipets, 5- and 10-mL, with pipet bulb.* 11. Volumetric flasks, 5- and 10-mL and 1-L.* 12. Plastic,* glass* or single element (e.g., tantalum) knives and forceps for cutting tissue samples, as needed [3,4,5]. 13. Work station with plastic work surfaces, with air cleaned by a vertical, laminar flow device and high efficiency particulate air filter or by electrostatic precipitator. 14. Gloves, plastic, metal-free. *All glassware and plasticware which contacts standards, blanks, or samples should be detergent-washed, thoroughly rinsed with tap and deionized water, soaked 12 h in 10% (v/v) HNO 3 and soaked 12 h in deionized water.
SPECIAL PRECAUTIONS: Concentrated acids are extremely corrosive; work with them only in a fume hood and wear appropriate safety equipment (safety glasses or face shield, gloves and labcoat). Samples of blood and tissue collected from humans pose a real health risk to laboratory workers who collect and handle these samples. These risks are primarily due to personal contact with infective biological samples and can have serious health consequences, such as infectious hepatitis, and other diseases. There is also some risk from the chemical content of these samples, but this is much less. Those who handle blood and tissue specimens should wear protective gloves, and avoid aerosolization of the samples. Mouth pipetting, of course, must be avoided.
SAMPLING: 1.
Collect blood samples in heparinized blood collection tubes. Mix immediately. Collect tissue samples (ca. 0.25 g for "dry" tissue or ca. 1 g for "wet" tissue) in bottles. NOTE: If unheparinized blood collection tubes are used, freeze samples immediately. NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94 ​ELEMENTS in blood or tissue: METHOD 8005, Issue 2, dated 15 August 1994 - Page 3 of 6 2.
Ship samples in dry ice and store frozen (<15 °C) prior to digestion.
SAMPLE PREPARATION: 3. 4. 5.
6. 7. 8.
Allow sample to equilibrate to room temperature. Transfer accurately weighed portions of 10 g blood, 0.25 g "dry" tissue, or 1.0 g "wet" tissue to a beaker. Add 10.0 mL digestion acid to each blood sample or 5.0 mL digestion acid to each tissue sample. Heat at 110 °C for 2 h. NOTE: Start reagent blanks, in triplicate, at this step. Increase hotplate temperature to 250 °C and heat until ca. 1 mL (for blood) or ca. 0.5 mL (for tissue) remains (2 to 3 h). Allow beaker to cool. Choose one of the following: a. External standard method. Transfer contents of beaker to a volumetric flask (10 mL for blood; 5 mL for tissue). Dilute to the mark with deionized water. b. Internal standard method. Add, via pipet, 10.0 mL (for blood) or 5.0 mL (for tissue) yttrium standard to the beaker.
CALIBRATION AND QUALITY CONTROL: 9. 10. 11.
Calibrate the spectrometer according to manufacturer's recommendations. NOTE: Typically, an acid blank and 10 µg/mL multielement solutions are used. Analyze a standard for every ten samples. Check measurement recoveries for all elements of interest with at least three spiked, unexposed samples or with reference materials of known elemental composition. These quality control samples should constitute 15 to 20% of all samples analyzed. NOTE: For blood or tissue spikes, split a control sample and spike one fraction. Subtract the element quantity found in the unspiked portion from the element quantity found in the spiked portion and determine measurement recovery. Correct all samples for this measurement recovery. This is especially important for blood samples because routine interelement corrections will not adequately compensate for the high levels of iron present, which has numerous spectral emission wavelengths.
MEASUREMENT: 12. 13.
Set the spectrometer to conditions specified by the manufacturer. Analyze standards and samples. NOTE: If the values for the samples are above the range of the standards, dilute the sample solutions with 10% H 2SO 4, reanalyze, and apply the appropriate dilution factor in calculations.
CALCULATIONS: 14. 15.
Obtain the solution concentration found for each element in the sample, C s (µg/mL), and the average blank, C b (µg/mL), from the measurement data. According to the method of standardization, calculate the concentration, C (µg/g), of each element in the mass of sample taken, M (g). a. External standard method. Use the final solution volumes of sample, V s (mL) and blank, Vb (mL): b. Internal standard method. Use the yttrium standard concentration, C y (µg/mL), the volume of yttrium standard added, V y (mL), and the concentration of yttrium found in the NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94 ​ELEMENTS in blood or tissue: METHOD 8005, Issue 2, dated 15 August 1994 - Page 4 of 6
sample, C sy (µg/mL). Treat the blank solution similarly [i.e., C b is the concentration (µg/mL) of analyte on the blank and C by is the concentration of yttrium in the blank]:
GUIDELINES TO INTERPRETATION: Acceptable and unacceptable levels for elements have not been determined by this method. Iyengar [6] reports metals concentrations in tissues and body fluids. Lauwerys [7] discusses metals and can be consulted for guidance and interpretation. For tissue, trace element concentrations will vary with the tissue type or organ. Iyengar [6] reports metals concentrations for different tissues of non-occupationally exposed people. Blood metal concentrations of non-occupationally exposed individuals are given in Table 2.
REFERENCES: [1] Hull, R. D. Analysis of Trace Metals for Occupationally Exposed Workers, Morbidity and Mortality Weekly Report, 33, (1984). [2] Hull, R. D. ICP-AES Multielement Analysis of Industrial Hygiene Samples, NTIS Publication No. PB 85-221414, (1985). [3] Katz, S. A. Amer. Biotechnology Lab., 2(4), 24-30 (1984). [4] Versieck, J., F. Barbier, R. Cornelis and J. Hoste. Talanta , 29, 973-984 (1982). [5] Behne, D. J. Clin. Chem. Clin. Biochem ., 19, 115-120 (1981). [6] Iyengar, G. V., W. E. Kollmer and H. J. M. Bowen. The Elemental Composition of Human Tissues and Body Fluids, Verlag Chemie., New York (1978). [7] Lauwerys, R. R. Industrial Chemical Exposure : Guidelines for Biological Monitoring , Biomedical Publications, Davis, CA (1983). [8] Toxic and Trace Metals in the Workplace and the Natural Environment, Environmental Sciences Associates, Inc., Bedford, MA 01730 (1981). [9] Tipton, I.H. and J.J. Shafer. Statistical Analysis of Lung Trace Element Levels, Arch. of Environ . Health, 8, 66 (1964). [10] Brune, D., G. Nordberg and P.O. Wester. Distribution of 23 Elements in the Kidney Liver and Lungs of Workers from a Smeltery and Refinery in North Sweden Exposed to a Number of Elements and of a Control Group., Sci. of the Total Environ ., 16, 13 (1980). [11] Mulay, I.L., R. Roy, B.E. Knox, N.H. Shur and W.E. Delaney. Trace Metal Analysis of Cancerous and Noncancerous Human Tissues, J. of the Natl. Cancer Inst., 47, 9 (1971). Some Inorganic Substances in Body Fluids and Tissues, presented at AIHA Annual Meeting, Detroit, MI (1961). [12] Smith, R.G. A Summary of Recent Information on "Normal Levels" and "Significant Levels" of Some Inorganic Substances in Body Fluids and Tissues, presented at AIHA Annual Meeting, Detroit, MI (1961). [13] Crable, J.V., R.G. Keenan, R.E. Kinser, A.W. Smallwood and P.A. Mauer. Metal and Mineral Concentrations in Lungs of Bituminous Coal Miners, Am. Ind. Hyg. Assoc. J., 29, 106 (1968). [14] Sweet, D.V., W.E. Crouse and J.V. Crable. Chemical and Statistical Studies of Contaminants in Urban Lungs, Am. Ind. Hyg. Assoc. J., 39, 515 (1978). [15] Lauwerys, R. Biological Criteria for Selected Industrial Toxic Chemicals: A Review, Scand. J. NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94 ​ELEMENTS in blood or tissue: METHOD 8005, Issue 2, dated 15 August 1994 - Page 5 of 6
[16]
[17] [18] [19]
Work Environ . and Health, 1, 139 (1975). Haas, W.H., K.W. Olson, V.A. Fassel and E.L. DeKalb. Development of Multielement Sampling and Analyses Methods Using Inductively Coupled Plasma-Atomic Emission Spectroscopy, Annual Progress Report for NIOSH, Interagency Agreement NIOSH-IA-77-24 (1980). Baselt, R.C. Biological Monitoring Methods for Industrial Chemicals , Biomedical Publications, Davis, CA (1980). Goldwater, L.J. Normal Concentrations of Metals in Urine and Blood, WHO Chron., 21(5), 191 (1967). Bowen, H.J.M. Trace Elements in Biochemistry , Academic Press (1966).
METHOD WRITTEN BY: R. DeLon Hull, Ph.D., NIOSH/DBBS.
TABLE 1. GENERAL INFORMATION Element (Formula) Antimony (Sb) Cadmium (Cd) Cobalt (Co) Chromium (Cr) Copper (Cu) Iron (Fe) Lanthanum (La) Lead (Pb) Lithium (Li) Magnesium (Mg) Manganese (Mn) Molybdenum (Mo) Nickel (Ni) Platinum (Pt) Silver (Ag) Strontium (Sr) Thallium (Tl) Vanadium (V) Zinc (Zn) Zirconium (Zr)
Atomic Weight
CAS#
RTECS
121.75 112.40 58.93 52.00 63.54 55.85 138.91 207.19 6.94 24.31 54.94 95.94 58.71 195.09 107.87 87.62 204.37 50.94 65.37 91.22
7440-36-0 7440-43-9 7440-48-4 7440-47-3 7440-50-8 7439-89-6 7439-91-0 7439-92-1 7439-93-2 7439-95-4 7439-96-5 7439-98-7 7440-02-0 7440-06-4 7440-22-4 7440-24-6 7440-28-0 7440-62-2 7440-66-6 7440-67-7
CC4025000 EU9800000 GF8750000 GB4200000 GL5325000 NO4565500 -OF7525000 OJ5540000 OM2100000 OO9275000 QA4680000 QR5950000 TP2160000 VW3500000 -XG3425000 YW1355000 ZG8600000 ZH7070000
NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94 ​ELEMENTS in blood or tissue: METHOD 8005, Issue 2, dated 15 August 1994 - Page 6 of 6 TABLE 2. RECOVERY OF METALS FROM BLOOD [1,2].
Element (Formula) Antimony (Sb) Cadmium (Cd) Cobalt (Co) Chromium (Cr) Copper (Cu) Iron (Fe) Lanthanum (La) Lead (Pb) Lithium (Li) Magnesium (Mg) Manganese (Mn) Molybdenum (Mo) Nickel (Ni) Platinum (Pt) Silver (Ag) Strontium (Sr) Thallium (Tl) Vanadium (V) Zinc (Zn) Zirconium (Zr)
Wavelength (nm) 217.58 226.5 231.2 205.6 324.8 45,000 -- c 220.4 670.8 279.6 257.6 281.6 231.6 203.7 328.3 421.5 190.9 310.2 213.9 339.2
Value a (µg/100 mL) 0.4 0.5 1.0 4.5 100 0 10 23 1.0 3,800 4.0 4.0 5.0 -- c 3.5 2.8 1.0 1.2 700 1.5
Metal "Nonexposed" Added (µg/Sample) 10 10 10 10 10 --b 119 10 10 110 10 10 10 10 10 10 10 10 60 10
Quantity Recovery (%) 106 120 81 114 101 -- b 2.4 113 113 104 98 126 86 92 115 113 97 131 103 71
a
Precision (% s r) n=4 4.9 1.1 21 4.7 5.8
0.85 1.1 12 2.1 3.1 16 14 0.8 0.88 8.7 1.1 17 8.7
"Nonexposed" value is the average concentration for the respective element in blood of non-occupationally exposed individuals. These values were tabulated from References [6] through [19]. b Recovery not determined (blood Fe concentration was above quantitation limit of spectrometer). c Concentration not reported.
NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94
Accuracy (±%) 15.6 22.2 60.2 23.2 12.4 -23.7 14.7 15.2 27.5 6.1 32.1 45.4 35.4 16.6 14.7 20.0 33.2 36.3 46.0