Analytical and Bioanalytical Chemistry

, Volume 410, Issue 22, pp 5689–5702 | Cite as

Speciation analysis of arsenic in seafood and seaweed: Part II—single laboratory validation of method

  • Mesay Mulugeta Wolle
  • Sean D. ConklinEmail author
Research Paper
Part of the following topical collections:
  1. Food Safety Analysis


Single laboratory validation of a method for arsenic speciation analysis in seafood and seaweed is presented. The method is based on stepwise extraction of water-soluble and non-polar arsenic with hot water and a mixture of dichloromethane and methanol, respectively. While the water-soluble arsenicals were speciated by anion and cation exchange liquid chromatography inductively coupled plasma mass spectrometry (LC-ICP-MS), the non-polar arsenicals were collectively determined by ICP-MS after digestion in acid. The performance characteristics and broad application of the method were evaluated by analyzing eight commercial samples (cod, haddock, mackerel, crab, shrimp, geoduck clam, oyster, and kombu) and four reference materials (fish protein (DORM-4), lobster hepatopancreas (TORT-3), mussel tissue (SRM 2976), and hijiki seaweed (CRM 7405-a)) representing finfish, crustaceans, molluscs, and seaweed. Matrices spiked at three levels in duplicates were also analyzed. The stepwise extraction provided 76–106% extraction of the total arsenic from the test materials. The method demonstrated satisfactory repeatability for analysis of replicate extracts prepared over several days. The accuracy of the method was evaluated by analyzing reference materials certified for both total arsenic and a few arsenicals; the experimental results were 90–105% of the certified values. Comparison between the total water-soluble arsenic and the sum of the concentrations of the chromatographed species gave 80–92% mass balance. While spike recoveries of most arsenicals were in the acceptance range set by CODEX, a few species spiked into cod, haddock, and shrimp were poorly recovered due to transformation to other forms. After thorough investigations, strategies were devised to improve the recoveries of these species by averting their transformations. Limits of quantification (LOQ) for the extraction and quantification of 16 arsenicals using the current method were in the range 6–16 ng g−1 arsenic.


Arsenic Seafood Seaweed Speciation Validation 



The authors thank Oak Ridge Institute for Science and Education (ORISE) for financial support. Sarah Stadig (Center for Food Safety and Applied Nutrition, FDA) is acknowledged for barcoding the seafood samples, and Dr. Kevin Kubachka (Forensic Chemistry Center, FDA) for kindly providing standards of arsenosugars.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_910_MOESM1_ESM.pdf (138 kb)
ESM 1 (PDF 138 kb)


  1. 1.
    Larsen R, Eilertsen K-E, Elvevoll EO. Health benefits of marine foods and ingredients. Biotechnol Adv. 2011;29:508–18.CrossRefPubMedGoogle Scholar
  2. 2.
    Molin M, Ulven SM, Meltzer HM, Alexander J. Arsenic in the human food chain, biotransformation and toxicology—review focusing on seafood arsenic. J Trace Elem Med Biol. 2015;31:249–59.CrossRefPubMedGoogle Scholar
  3. 3.
    Francesconi KA. Arsenic species in seafood: origin and human health implication. Pure Appl Chem. 2010;82:373–81.CrossRefGoogle Scholar
  4. 4.
    Taylor V, Goodale B, Raab A, Schwerdtle T, Reimer K, Conklin S, et al. Human exposure to organic arsenic species from seafood. Sci Total Environ. 2017;580:266–82.CrossRefPubMedGoogle Scholar
  5. 5.
    Kaise T, Watanabe S, Itoh K. The acute toxicity of arsenobetaine. Chemosphere. 1985;14:1327–32.CrossRefGoogle Scholar
  6. 6.
    Feldmann J, Krupp EM. Critical review or scientific opinion paper: arsenosugars—a class of benign arsenic species or justification for developing partly speciated arsenic fractionation in foodstuffs? Anal Bioanal Chem. 2011;399:1735–41.CrossRefPubMedGoogle Scholar
  7. 7.
    World Health Organization, International Agency for Research on Cancer (IARC Monographs 100), A review of human carcinogens. C. Metals, arsenic, dusts and fibres, 2012, Lyon, France.Google Scholar
  8. 8.
    Francesconi KA, Tanggaard R, McKenzie CJ, Goessler W. Arsenic metabolites in human urine after ingestion of an arsenosugar. Clin Chem. 2002;48:92–101.PubMedGoogle Scholar
  9. 9.
    Hulle M, Zhang C, Schotte B, Mees L, Vanhaecke F, Vanholder R, et al. Identification of some arsenic species in human urine and blood after ingestion of Chinese seaweed Laminaria. J Anal At Spectrom. 2004;19:58–64.CrossRefGoogle Scholar
  10. 10.
    Andrewes P, Demarini DM, Funasaka K, Wallace K, Lai VWM, Sun H, et al. Do arsenosugars pose a risk to human health? The comparative toxicities of a trivalent and pentavalent arsenosugar. Environ Sci Technol. 2004;38:4140–8.CrossRefPubMedGoogle Scholar
  11. 11.
    Schmeisser E, Goessler W, Francesconi KA. Human metabolism of arsenolipids present in cod liver. Anal Bioanal Chem. 2006;385:367–76.CrossRefPubMedGoogle Scholar
  12. 12.
    Meyer S, Matissek M, Muller SM, Taleshi MS, Ebert F, Francesconi KA, et al. In vitro toxicological characterization of three arsenic-containing hydrocarbons. Metallomics. 2014;6:1023–33.CrossRefPubMedGoogle Scholar
  13. 13.
    Sele V, Sloth JJ, Lundebye A-K, Larsen EH, Berntssen MHG, Amlund H. Arsenolipids in marine oils and fats: a review of occurrence, chemistry and future research needs. Food Chem. 2012;133:618–30.CrossRefGoogle Scholar
  14. 14.
    Francesconi KA, Kuehnelt D. Determination of arsenic species: a critical review of methods and applications, 2000–2003. Analyst. 2004;129:373–95.CrossRefPubMedGoogle Scholar
  15. 15.
    Maher WA, Ellwood MJ, Krikowa F, Raber G, Foster S. Measurement of arsenic species in environmental, biological fluids and food samples by HPLC-ICPMS and HPLC-HG-AFS. J Anal At Spectrom. 2015;30:2129–83.CrossRefGoogle Scholar
  16. 16.
    Larsen EH, Engman J, Sloth JJ, Hansen M, Jorhem L. Determination of inorganic arsenic in white fish using microwave-assisted alkaline alcoholic sample dissolution and HPLC-ICP-MS. Anal Bioanal Chem. 2005;381:339–46.CrossRefPubMedGoogle Scholar
  17. 17.
    Pétursdóttir ÁH, Gunnlaugsdóttir H, Krupp EM, Feldmann J. Inorganic arsenic in seafood: does the extraction method matter? Food Chem. 2014;150:353–9.CrossRefPubMedGoogle Scholar
  18. 18.
    Wahlen R, McSheehy S, Scriver C, Mester Z. Arsenic speciation in marine certified reference materials part 2. The quantification of water-soluble arsenic species by high-performance liquid chromatography-inductively coupled plasma mass spectrometry. J Anal At Spectrom. 2004;19:876–82.CrossRefGoogle Scholar
  19. 19.
    Ackley KL, B’Hymer C, Sutton KL, Caruso JA. Speciation of arsenic in fish tissue using microwave-assisted extraction followed by HPLC-ICP-MS. J Anal At Spectrom. 1999;14:845–50.CrossRefGoogle Scholar
  20. 20.
    Kuehnelt D, Irgolic KJ, Goessler W. Comparison of three methods for extraction of arsenic compounds from NRCC standard reference material DORM-2 and the brown alga Hijiki fuziforme. Appl Organomet Chem. 2001;15:445–56.CrossRefGoogle Scholar
  21. 21.
    Reyes LH, Mar JLG, Rahman GMM, Seybert B, Fahrenholz T, Kingston HM. Simultaneous determination of arsenic and selenium species in fish tissues using microwave-assisted enzymatic extraction and ion chromatography–inductively coupled plasma mass spectrometry. Talanta. 2009;78:983–90.CrossRefPubMedGoogle Scholar
  22. 22.
    Leufroy A, Noël L, Dufailly V, Beauchemin D, Guérin T. Determination of seven arsenic species in seafood by ion exchange chromatography coupled to inductively coupled plasma-mass spectrometry following microwave assisted extraction: method validation and occurrence data. Talanta. 2011;83:770–9.CrossRefPubMedGoogle Scholar
  23. 23.
    Zmozinski AV, Llorente-Mirandes T, López-Sánchez JF, da Silva MM. Establishment of a method for determination of arsenic species in seafood by LC-ICP-MS. Food Chem. 2015;173:1073–82.CrossRefPubMedGoogle Scholar
  24. 24.
    Wolle MM, Conklin SD. Speciation analysis of arsenic in seafood and seaweed: part I—evaluation and optimization of methods. Anal Bioanal Chem.
  25. 25.
    US FDA Office of Foods and Veterinary Medicine, Guidelines for validation of chemical methods for the FDA Foods and Veterinary Medicine Program, 2nd ed., April 2015 (
  26. 26.
    US FDA Elemental Analysis Manual for Food and Related Products, Section 4.7. Inductively Coupled Plasma-Mass Spectrometric Determination of Arsenic, Cadmium, Chromium, Lead, Mercury, and Other Elements in Food Using Microwave Assisted Digestion, Version 1.1, March 2015 (
  27. 27.
    US FDA Elemental Analysis Manual for Food and Related Products, Section 3.2. Terminology, September 2015 (
  28. 28.
    Sloth JJ, Larsen EH, Julshamn K. Selective arsenic speciation analysis of human urine reference materials using gradient elution ion-exchange HPLC-ICP-MS. J Anal At Spectrom. 2004;19:973–8.CrossRefGoogle Scholar
  29. 29.
    Codex Alimentarius Commission, Procedural Manual, 24th ed., 2015, Rome, Italy.Google Scholar
  30. 30.
    US FDA Elemental Analysis Manual for Food and Related Products, Section 4.10. High Performance Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometric Determination of Four Arsenic Species in Fruit Juice, Version 1.0, July 2013 (
  31. 31.
    US FDA Elemental Analysis Manual for Food and Related Products, Section 4.11. Arsenic Speciation in Rice and Rice Products Using High Performance Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometric Determination, Version Draft 1.1, November 2012 (
  32. 32.
    Rey NA, Howarth OW, Pereira-Maia EC. Equilibrium characterization of the as(III)–cysteine and the as(III)–glutathione systems in aqueous solution. J Inorg Biochem. 2004;98:1151–9.CrossRefPubMedGoogle Scholar
  33. 33.
    Jiang G, Gong Z, Li X-F, Cullen WR, Le XC. Interaction of trivalent arsenicals with metallothioneins. Chem Res Toxicol. 2003;16:873–80.CrossRefPubMedGoogle Scholar
  34. 34.
    Lu M, Wang H, Li X-F, Lu X, Cullen WR, Arnold LL, et al. Evidence of hemoglobin binding to arsenic as a basis for the accumulation of arsenic in rat blood. Chem Res Toxicol. 2004;17:1733–42.CrossRefPubMedGoogle Scholar
  35. 35.
    Wang Z, Zhang H, Li X-F, Le XC. Study of interactions between arsenicals and thioredoxins (human and E. coli) using mass spectrometry. Rapid Commun Mass Spectrom. 2007;21:3658–66.CrossRefPubMedGoogle Scholar
  36. 36.
    Winther JR, Thorpe C. Quantification of thiols and disulfides. Biochim Biophys Acta. 1840;2014:838–46.Google Scholar
  37. 37.
    Llorente-Mirandes T, Calderón J, López-Sánchez JF, Centrich F, Rubio R. A fully validated method for the determination of arsenic species in rice and infant cereal products. Pure Appl Chem. 2012;84:225–38.CrossRefGoogle Scholar
  38. 38.
    Ruttens A, Blanpain AC, De Temmerman L, Waegeneers N. Arsenic speciation in food in Belgium, part 1: fish, molluscs and crustaceans. J Geochem Explor. 2012;121:55–61.CrossRefGoogle Scholar
  39. 39.
    Cao X, Hao C, Wang G, Yang H, Chen D, Wang X. Sequential extraction combined with HPLC–ICP-MS for As speciation in dry seafood products. Food Chem. 2009;113:720–6.CrossRefGoogle Scholar
  40. 40.
    Amayo KO, Raab A, Krupp EM, Marschall T, Horsfall M Jr, Feldmann J. Arsenolipids show different profiles in muscle tissues of four commercial fish species. J Trace Elem Med Biol. 2014;28:131–7.CrossRefPubMedGoogle Scholar
  41. 41.
    Glabonjat RA, Raber G, Jensen KB, Ehgartner J, Francesconi KA. Quantification of arsenolipids in the certified reference material NMIJ 7405-a (Hijiki) using HPLC/mass spectrometry after chemical derivatization. Anal Chem. 2014;86:10282–7.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Argese E, Bettiol C, Rigo C, Bertini S, Colomban S, Ghetti PF. Distribution of arsenic compounds in Mytilus galloprovincialis of the Venice lagoon (Italy). Sci Total Environ. 2005;348:267–77.CrossRefPubMedGoogle Scholar

Copyright information

© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2018

Authors and Affiliations

  1. 1.Division of Bioanalytical Chemistry, Office of Regulatory Science, Center for Food Safety and Applied NutritionUS Food and Drug AdministrationCollege ParkUSA

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