Analytical and Bioanalytical Chemistry

, Volume 404, Issue 1, pp 89–99 | Cite as

Multi-elemental bio-imaging of rat tissue from a study investigating the bioavailability of bismuth from shotgun pellets

  • Dagmar S. Urgast
  • Dag G. Ellingsen
  • Balázs Berlinger
  • Einar Eilertsen
  • Grete Friisk
  • Vidar Skaug
  • Yngvar Thomassen
  • John H. Beattie
  • In-Sook Kwun
  • Jörg FeldmannEmail author
Original Paper


In recent years, bismuth has been promoted as a “green element” and is used as a substitute for the toxic lead in ammunition and other applications. However, the bioavailability and toxicity of bismuth is still not very well described. Following a hunting accident with bismuth-containing shots, a bioavailability study of bismuth from metal pellets inoculated into rat limb muscles was carried out. Bismuth could be found in urine and blood of the animals. Bio-imaging using laser ablation ICP-MS of thin sections of the tissue around the metal implant was carried out to find out more about the distribution of the metal diffusing into the tissue. Two laser ablation systems with different ablation cell designs were compared regarding their analytical performance. Low concentrations of bismuth showing a non-symmetrical pattern were detected in the tissue surrounding the metal implant. This was partly an artefact from cutting the thin sections but also bio-mobilisation of the metals of the implant could be seen. An accumulation of zinc around the implant was interpreted as a marker of inflammation. Challenges regarding sample preparation for laser ablation and bio-imaging of samples of diverse composition became apparent during the analysis.


Bismuth Bio-imaging LA-ICP-MS Tissue thin section Rat 



The laser ablation work was supported by the Global Research Network Program of the National Research Foundation of South Korea (NRF-2008-220-F00013), the TESLA Research Fund, and the School of Computing and Natural Sciences of the University of Aberdeen.


  1. 1.
    Carlin JF (2008) In: U.S. Geological Survey Minerals Yearbook, p 121Google Scholar
  2. 2.
    Mahony D, Woods A, Eelman M et al (2005) Interaction of bismuth subsalicylate with fruit juices, ascorbic acid, and thiol-containing substrates to produce soluble bismuth products active against Clostridium difficile. Antimicrob Agents Chemother 49:431CrossRefGoogle Scholar
  3. 3.
    Lambertucci SA, Donazar JA, Hiraldo F (2010) Poisoning people and wildlife with lead ammunition: time to stop. Environ Sci Technol 44:7759CrossRefGoogle Scholar
  4. 4.
    UNEP/AEWA report 2009: Phasing out the use of lead shots for hunting in wetlands - experiences made and lessons learned by AEWA range statesGoogle Scholar
  5. 5.
    Fahey NSC, Tsuji LJS (2006) Is there a need to re-examine the approval of bismuth shotshell as a non-toxic alternative to lead based on the precautionary principle? J Environ Monit 8:1190CrossRefGoogle Scholar
  6. 6.
    Feldmann J, Krupp E, Glindemann D et al (1999) Methylated bismuth in the environment. Appl Organomet Chem 13:739CrossRefGoogle Scholar
  7. 7.
    Michalke K, Meyer J, Hirner A et al (2002) Biomethylation of bismuth by the methanogen Methanobacterium formicicum. Appl Organomet Chem 16:221CrossRefGoogle Scholar
  8. 8.
    Tillman L, Drake F, Dixon J et al (1996) Review article: safety of bismuth in the treatment of gastrointestinal diseases. Aliment Pharmacol Ther 10:459CrossRefGoogle Scholar
  9. 9.
    Hollmann M, Boertz J, Dopp E et al (2010) Parallel on-line detection of a methylbismuth species by hyphenated GC/EI-MS/ICP-MS technique as evidence for bismuth methylation by human hepatic cells. Metallomics 2:52CrossRefGoogle Scholar
  10. 10.
    von Recklinghausen U, Hartmann LM, Rabieh S et al (2008) Methylated bismuth, but not bismuth citrate or bismuth glutathione, induces cyto- and genotoxic effects in human cells in vitro. Chem Res Toxicol 21:1219CrossRefGoogle Scholar
  11. 11.
    Tsuji L, Wainman B, Jayasinghe R et al (2004) Tissue-bismuth levels of game birds harvested with bismuth shotshell: policy implications. Bull Environ Contam Toxicol 72:128CrossRefGoogle Scholar
  12. 12.
    Pamphlett R, Danscher G, Rungby J et al (2000) Tissue uptake of bismuth from shotgun pellets. Environ Res 82:258CrossRefGoogle Scholar
  13. 13.
    Stoltenberg M, Locht L, Larsen A et al (2003) In vivo cellular uptake of bismuth ions from shotgun pellets. Histol Histopathol 18:781Google Scholar
  14. 14.
    Jackson B, Harper S, Smith L et al (2006) Elemental mapping and quantitative analysis of Cu, Zn, and Fe in rat brain sections by laser ablation ICP-MS. Anal Bioanal Chem 384:951CrossRefGoogle Scholar
  15. 15.
    Zoriy MV, Dehnhardt M, Matusch A et al (2008) Comparative imaging of P, S, Fe, Cu, Zn and C in thin sections of rat brain tumor as well as control tissues by laser ablation inductively coupled plasma mass spectrometry. Spectrochim Acta B Atomic Spectrosc 63:375CrossRefGoogle Scholar
  16. 16.
    Hare D, Reedy B, Grimm R et al (2009) Quantitative elemental bio-imaging of Mn, Fe, Cu and Zn in 6-hydroxydopamine induced Parkinsonism mouse models. Metallomics 1:53CrossRefGoogle Scholar
  17. 17.
    Corbin BD, Seeley EH, Raab A et al (2008) Metal chelation and inhibition of bacterial growth in tissue abscesses. Science 319:962CrossRefGoogle Scholar
  18. 18.
    Kindness A, Sekaran C, Feldmann J (2003) Two-dimensional mapping of copper and zinc in liver sections by laser ablation-inductively coupled plasma mass spectrometry. Clin Chem 49:1916CrossRefGoogle Scholar
  19. 19.
    Feldmann J, Kindness A, Ek P (2002) Laser ablation of soft tissue using a cryogenically cooled ablation cell. J Anal At Spectrom 17:813CrossRefGoogle Scholar
  20. 20.
    McRae R, Bagchi P, Sumalekshmy S et al (2009) In situ imaging of metals in cells and tissues. Chem Rev 109:4780CrossRefGoogle Scholar
  21. 21.
    Urgast DS, Ou O, Gordon M et al (2012) Microanalytical isotope ratio measurements and elemental mapping using laser ablation ICP-MS for tissue thin sections: zinc tracer studies in rats. Anal Bioanal Chem 402:287CrossRefGoogle Scholar
  22. 22.
    Zalewski P, Truong-Tran A, Grosser D et al (2005) Zinc metabolism in airway epithelium and airway inflammation: basic mechanisms and clinical targets. A review. Pharmacol Ther 105:127CrossRefGoogle Scholar
  23. 23.
    Luster A (1998) Chemokines—chemotactic cytokines that mediate inflammation. N Engl J Med 338:436CrossRefGoogle Scholar
  24. 24.
    Austin C, Fryer F, Lear J et al (2011) Factors affecting internal standard selection for quantitative elemental bio-imaging of soft tissues by LA-ICP-MS. J Anal At Spectrom 26:1494CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Dagmar S. Urgast
    • 1
  • Dag G. Ellingsen
    • 2
  • Balázs Berlinger
    • 2
  • Einar Eilertsen
    • 2
  • Grete Friisk
    • 2
  • Vidar Skaug
    • 2
  • Yngvar Thomassen
    • 2
  • John H. Beattie
    • 3
  • In-Sook Kwun
    • 4
  • Jörg Feldmann
    • 1
    Email author
  1. 1.TESLA (Trace Element Speciation Laboratory)University of Aberdeen, College of Physical Sciences, Department of ChemistryAberdeenUK
  2. 2.National Institute of Occupational HealthOsloNorway
  3. 3.Rowett Institute of Nutrition and Health, College of Life Sciences and Medicine, Micronutrients GroupUniversity of AberdeenAberdeenUK
  4. 4.Department of Food Science and NutritionAndong National UniversityAndongRepublic of Korea

Personalised recommendations