Skip to main content

Development of mass spectrometric methods for detecting arsenic-glutathione complexes

Abstract

It has been suggested recently that arsenic-glutathione (As-GSH) complexes play an important role in the methylation of arsenic. The present study describes the development of high-performance liquid chromatography (HPLC)-electrospray tandem mass spectrometry (ES-MS/MS), operated in the selected reaction monitoring (SRM) mode, and HPLC-inductively coupled plasma mass spectrometry (ICP-MS) methods suitable for the sensitive and selective identification of four As-GSH complexes. Method optimization was carried out using a series of synthetically prepared standards, i.e., three As-GSH species containing trivalent arsenic: tri(glutamyl-cysteinyl-glycinyl)trithio-arsenite (ATG), di(glutamyl-cysteinyl-glycinyl)methyl-dithio-arsonite (MADG), and (ã-glutamyl-cysteinyl-glycinyl) dimethyl-thio-arsinite (DMAG), as well as one As-GSH species containing pentavalent As: dimethylthioarsinic acid-glutathione (DMTAV-GSH). The collision induced dissociation behavior of these compounds was investigated in detail to identify optimum SRM transitions for each complex. Both methods were based on reversed-phase chromatography using gradient elution with methanol, formic acid, and water as solvents. The amount of methanol that was used with this HPLC method (up to 12% vol/vol) was compatible with ICP-MS, without the need of a specially adapted interface. Subsequently, these analytical methods were applied to carry out a preliminary investigation about the role of As-GSH complexes in the methylation of arsenite by methylcobalamin (CH3B12) in the presence of glutathione (GSH). For the first time, the complexes ATG, MADG, and trace amounts of DMAG were detected as products of this reaction.

References

  1. Mandal, B. K.; Suzuki, K. T. Arsenic Round the World: A Review. Talanta 2002, 58, 201–235.

    CAS  Article  Google Scholar 

  2. Smith, A. H.; Hopenhayn-Rich, C.; Bates, M. N.; Goeden, H. M.; Hertz-Picciotto, I.; Duggan, H. M.; Wood, R.; Kosnett, M. J.; Smith, M. T. Cancer Risks from Arsenic in Drinking Water. Environ. Health Perspect. 1992, 97, 259–267.

    CAS  Article  Google Scholar 

  3. Suzuki, K. T.; Mandal, B. K.; Katagiri, A.; Sakuma, Y.; Kawakami, A.; Ogra, Y.; Yamaguchi, K.; Sei, Y.; Yamanaka, K.; Anzai, K.; Ohmichi, M.; Takayama, H.; Aimi, N. Dimethylthioarsenicals as Arsenic Metabolites and Their Chemical Preparations. Chem. Res. Toxicol. 2004, 17, 914–921.

    CAS  Article  Google Scholar 

  4. Le, X. C.; Lu, X.; Ma, M.; Cullen, W. R.; Aposhian, H. V.; Zheng, B. Speciation of Key Arsenic Metabolic Intermediates in Human Urine. Anal. Chem. 2000, 72, 5172–5177.

    CAS  Article  Google Scholar 

  5. Challenger, F. Biological methylation. Chem. Rev. 1945, 36, 315–361.

    CAS  Article  Google Scholar 

  6. Hayakawa, T.; Kobayashi, Y.; Cui, X.; Hirano, S. A. New Metabolic Pathway of Arsenite: Arsenic-Glutathione Complexes are Substrates for Human Arsenic Methyltransferase Cyt19. Arch. Toxicol. 2005, 79, 183–191.

    CAS  Article  Google Scholar 

  7. Hansen, H. R.; Pickford, R.; Thomas-Oates, J.; Jaspars, M.; Feldmann, J. 2-Dimethylarsinothioyl Acetic Acid Identified in a Biological Sample: The First Occurrence of a Mammalian Arsinothioyl Metabolite. Angew. Chem. Int. Ed. 2004, 116, 341–344.

    Article  Google Scholar 

  8. Schmeisser, E.; Raml, R.; Francesconi, K. A.; Kuehnelt, D.; Lindberg, A.-L.; Sörös, C.; Goessler, W. Thio Arsenosugars Identified as Natural Constituents of Mussels by Liquid Chromatography Mass Spectrometry. Chem. Commun. 2004, 16, 1824–1825.

    Article  Google Scholar 

  9. Fricke, M. W.; Creed, P. A.; Parks, A. N.; Shoemaker, J. A.; Schwegel, C. A.; Creed, J. T. Extraction and Detection of a New Arsine Sulfide Containing Arsenosugar in Mollusks by IC-ICP-MS and IC-ESI-MS/MS. J. Anal. At. Spectrom. 2004, 19, 1454–1459.

    CAS  Article  Google Scholar 

  10. Nischwitz, V.; Kanaki, K.; Pergantis, S. A. Mass Spectrometric Identification of Novel Arsinothioyl-sugars in Marine Bivalves and Algae. J. Anal. At. Spectrom. 2006, 21, 33–40.

    CAS  Article  Google Scholar 

  11. Raab, A.; Schat, H.; Meharg, A. A.; Feldmann, J. Uptake, Translocation and Transformation of Arsenate and Arsenite in Sunflower (Helianthus annuus): Formation of Arsenic-Phytochelatin Complexes During Exposure to High Arsenic Concentrations. New Phytologist 2005, 168, 551–558.

    CAS  Article  Google Scholar 

  12. Kala, S. V.; Neely, M. W.; Kala, G.; Prater, C. I.; Atwood, D. W.; Rice, J. S.; Lieberman, M. W. The MRP2/cMOAT Transporter and Arsenic-Glutathione Complex Formation are Required for Biliary Excretion of Arsenic. J. Biol. Chem. 2000, 275, 33404–33408.

    CAS  Article  Google Scholar 

  13. Kobayashi, Y.; Cui, X.; Hirano, S. Stability of Arsenic Metabolites, Arsenic Triglutathione [As(GS)3] and Methylarsenic Diglutathione [CH3As(GS)2] in Rat Bile. Toxicology 2005, 211, 115–123.

    CAS  Article  Google Scholar 

  14. Cui, X.; Kobayashi, Y.; Hayakawa, T.; Hirano, S. Arsenic Speciation in Bile and Urine Following Oral and Intravenous Exposure to Inorganic and Organic Arsenics in Rats. Toxicol. Sci. 2004, 82, 478–487.

    CAS  Article  Google Scholar 

  15. Kala, S. V.; Kala, G.; Prater, C. I.; Sartorelli, A. C.; Lieberman, M. W. Formation and Urinary Excretion of Arsenic Triglutathione and Methylarsenic Diglutathione. Chem. Res. Toxicol. 2004, 17, 243–249.

    CAS  Article  Google Scholar 

  16. Raab, A.; Wright, S. H.; Jaspars, M.; Meharg, A. A.; Feldmann, J. Pentavalent Arsenic Can Bind to Biomolecules. Angew. Chem. Int. Ed. 2007, 46, 2594–2597.

    CAS  Article  Google Scholar 

  17. Raab, A.; Meharg, A. A.; Jaspars, M.; Genneyb, D. R.; Feldmann, J. Arsenic-Glutathione Complexes—Their Stability in Solution and During Separation by Different HPLC Modes. J. Anal. At. Spectrom. 2004, 19, 183–190.

    CAS  Article  Google Scholar 

  18. Scott, N.; Hatlelid, K. M.; MacKenzie, N. E.; Carter, D. E. Reactions of Arsenic(III) and Arsenic(V) Species with Glutathione. Chem. Res. Toxicol. 1993, 6, 102–106.

    CAS  Article  Google Scholar 

  19. Delnomdedieu, M.; Basti, M. M.; Otvos, J. D.; Thomas, D. J. Reduction and Binding of Arsenate and Dimethylarsinate by Glutathione: A Magnetic Resonance Study. Chem. Biol. Interact. 1994, 90, 139–155.

    CAS  Article  Google Scholar 

  20. Styblo, M.; Yamauchi, H.; Thomas, D. Comparative in Vitro Methylation of Trivalent and Pentavalent Arsenicals. Toxicol. Appl. Pharmacol. 1995, 135, 172–178.

    CAS  Article  Google Scholar 

  21. Styblo, M.; Thomas, D. J. In Vitro Inhibition of Glutathione Reductase by Arsenotriglutathione. Biochem. Pharmacol. 1995, 49, 971–977.

    CAS  Article  Google Scholar 

  22. Burford, N.; Eelman, M. D.; Groom, K. Identification of Complexes Containing Glutathione with As(III), Sb(III), Cd(II), Hg(II), Tl(I), Pb(II), or Bi(III) by Electrospray Ionization Mass Spectrometry. Inorg. Biochem. 2005, 99, 1992–1997.

    CAS  Article  Google Scholar 

  23. Zakharyan, R. A.; Aposhian, H. V. Arsenite Methylation by Methylvitamin B12 and Glutathione Does Not Require an Enzyme. Toxicol. Appl. Pharmacol. 1999, 154, 287–291.

    CAS  Article  Google Scholar 

  24. Pergantis, S. A.; Miguens-Rodriguez, M.; Vela, N. P.; Heitkemper, D. T. Investigating the Nonenzymatic Methylation of Arsenite by Methylcobalamin B12 using High-Performance Liquid Chromatography On-Line with Inductively Coupled Plasma-Mass Spectrometry. J. Anal. At. Spectrom. 2004, 19, 178–182.

    CAS  Article  Google Scholar 

  25. Nischwitz, V.; Pergantis, S. A. Liquid Chromatography On-Line with Selected Reaction Monitoring Electrospray Mass Spectrometry for the Determination of Organoarsenic Species in Crude Extracts of Marine Reference Materials. Anal. Chem. 2005, 77, 5551–5563.

    CAS  Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Spiros A. Pergantis.

Additional information

Published online June 28, 2008

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kanaki, K., Pergantis, S.A. Development of mass spectrometric methods for detecting arsenic-glutathione complexes. J Am Soc Mass Spectrom 19, 1559–1567 (2008). https://doi.org/10.1016/j.jasms.2008.06.011

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1016/j.jasms.2008.06.011

Keywords

  • Arsenite
  • Collision Induce Dissociation
  • Select Reaction Monitoring
  • Tandem Mass Spectrum
  • Sodium Arsenite