Analytical and Bioanalytical Chemistry

, Volume 410, Issue 22, pp 5703–5710 | Cite as

Inter-laboratory validation of an inexpensive streamlined method to measure inorganic arsenic in rice grain

  • Rufus L. ChaneyEmail author
  • Carrie E. Green
  • Steven J. Lehotay
Research Paper
Part of the following topical collections:
  1. Food Safety Analysis


With the establishment by CODEX of a 200 ng/g limit of inorganic arsenic (iAs) in polished rice grain, more analyses of iAs will be necessary to ensure compliance in regulatory and trade applications, to assess quality control in commercial rice production, and to conduct research involving iAs in rice crops. Although analytical methods using high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) have been demonstrated for full speciation of As, this expensive and time-consuming approach is excessive when regulations are based only on iAs. We report a streamlined sample preparation and analysis of iAs in powdered rice based on heated extraction with 0.28 M HNO3 followed by hydride generation (HG) under control of acidity and other simple conditions. Analysis of iAs is then conducted using flow-injection HG and inexpensive ICP-atomic emission spectroscopy (AES) or other detection means. A key innovation compared with previous methods was to increase the acidity of the reagent solution with 4 M HCl (prior to reduction of As5+ to As3+), which minimized interferences from dimethylarsinic acid. An inter-laboratory method validation was conducted among 12 laboratories worldwide in the analysis of six shared blind duplicates and a NIST Standard Reference Material involving different types of rice and iAs levels. Also, four laboratories used the standard HPLC-ICP-MS method to analyze the samples. The results between the methods were not significantly different, and the Horwitz ratio averaged 0.52 for the new method, which meets official method validation criteria. Thus, the simpler, more versatile, and less expensive method may be used by laboratories for several purposes to accurately determine iAs in rice grain.

Graphical abstract

Comparison of iAs results from new and FDA methods


Inorganic arsenic (iAs) analysis Rice Inter-laboratory validation Hydride generation (HG) Inductively coupled plasma-atomic emission spectroscopy (ICP-AES) ICP-MS 



We thank Anna McClung for providing the rice samples grown with traditional flood irrigation vs. alternative wetting and drying irrigation for the study. We also thank Amy Poet and Julie Napolitano for their extensive and careful work which was critical to the project. We very much appreciate the scientists and their laboratory colleagues who participated in the inter-laboratory validation study: (analysis by HG-ICP-AES) Amy Poet and Julie Napolitano, Adaptive Cropping Systems Laboratory, USDA-ARS, Beltsville, MD, USA; Tomohito Arao and Koji Baba, National Institute for Agro-Environmental Sciences, Tskuba, Ibaraki, Japan; Philip Moore, USDA-ARS, University of Arkansas, Fayetteville, AR, USA; Trenton Roberts, Dept. of Crop, Soil, and Environmental Sciences, Fayetteville, AR, USA; Eri Matsumoto, Tama Laboratory, Japan Food Research Laboratories, Tokyo, Japan; (by HG-AFS) Fangjie Zhao, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, China; (by HG-ICP-MS) Michael Bukowski (and by HG-AAS), USDA-ARS-GFHNRL, Grand Forks, ND, USA; Jörg Feldmann and Andrea Raab (by HPLC-ICP-MS), Trace Element Speciation Laboratory, University of Aberdeen, Scotland, UK; Anitha Kunhikrishnan, Won-Il Kim, and Ji-Hyock Yoo, Dept. of Agro-Food Safety, National Institute of Agricultural Sciences, Republic of Korea; Cheryl Stephenson and Marvin Boyd, Jr., Eurofins Central Analytical Laboratories, New Orleans, LA, USA; Yong-Guan Zhu and Guo-Xin Sun, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, China; (by HPLC-ICP-MS) Sean Conklin, Chemical Contaminants Branch, US Food and Drug Administration, Center for Food Safety and Applied Nutrition, College Park, MD, USA; Kent Lanclos and James Krest, Technology and Science Division, USDA Grain Inspection, Packers, and Stockyards Administration, National Grain Center, Kansas City, MO, USA; and Andrew Meharg and Manus Carey, Institute for Global Food Security, Queen’s University Belfast, Northern Ireland, UK.

Funding information

Funding for this research was provided by The Rice Foundation.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1075_MOESM1_ESM.pdf (497 kb)
ESM 1 (PDF 496 kb)


  1. 1.
    Codex Alimentarius. 2017a. General standard for contaminants and toxins in food and feed. CODEX STAN 193–1995. (CSX_193e.pdf).Google Scholar
  2. 2.
    D’Amato M, Forte G, Caroli S. Identification and quantification of major species of arsenic in rice. J AOAC Int. 2004;87:238–43.PubMedGoogle Scholar
  3. 3.
    de la Calle MB, Emteborg H, Linsinger TPJ, Montoro R, Sloth JJ, Rubio R, et al. Does the determination of inorganic arsenic in rice depend on the method? Trends Anal Chem. 2011;30:641–51.CrossRefGoogle Scholar
  4. 4.
    Heitkemper DT, Vela NP, Stewart KR, Westphal CS. Determination of total and speciated arsenic in rice by ion chromatography and inductively coupled plasma mass spectrometry. J Anal Atom Spectr. 2001;16:299–306.CrossRefGoogle Scholar
  5. 5.
    Heitkemper DT, Kubachka KM, Halpin PR, Allen MN, Shockey NV. Survey of total arsenic and arsenic speciation in US-produced rice as a reference point for evaluating change and future trends. Food Addit Contam B. 2009;2:112–20.CrossRefGoogle Scholar
  6. 6.
    Kubachka KM, Shockey NV, Hanley TA, Conklin SD, Heitkemper DT. Elemental analysis manual Section 4.11: Arsenic speciation in rice and rice products using high performance liquid chromatography-inductively coupled plasma-mass spectrometric determination. Version 1.1, 2012. (
  7. 7.
    Llorente-Mirandes T, Calderon J, Lopez-Sanchez 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
  8. 8.
    European Food Safety Agency. Scientific opinion on arsenic in food. EFSA J. 2009;7:1351.CrossRefGoogle Scholar
  9. 9.
    Jackson BP. Fast ion chromatography-ICP-QQQ for arsenic speciation. J Anal Atom Spectrom. 2015;30:1405–7.CrossRefGoogle Scholar
  10. 10.
    Gray PJ, Tanaba CK, Ebeler SE, Nelson J. A fast and fit-to-purpose arsenic speciation method for wine and rice. J Anal Atom Spectrom. 2017;32:1031–4.CrossRefGoogle Scholar
  11. 11.
    Narukawa T, Chiba K, Sinaviwat S, Feldmann J. A rapid monitoring method for inorganic arsenic in rice flour using reversed phase - high performance liquid chromatography - inductively coupled plasma mass spectrometry. J Chromatogr A. 2017;1479:129–36.CrossRefPubMedGoogle Scholar
  12. 12.
    Onken BM, Hossner LR. Determination of arsenic species in soil solution under flooded conditions. Soil Sci Soc Am J. 1996;60:1385–92.CrossRefGoogle Scholar
  13. 13.
    Musil S, Pétursdóttir ÁH, Raab A, Gunnlaugsdóttir H, Krupp E, Feldmann J. Speciation without chromatography using selective hydride generation: inorganic arsenic in rice and samples of marine origin. Anal Chem. 2014;86:993–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Pétursdóttir ÁH, Friedrich N, Musil S, Raab A, Gunnlaugsdóttir H, Krupp EM, et al. Hydride generation ICP-MS as a simple method for determination of inorganic arsenic in rice for routine biomonitoring. Anal Methods. 2014;6:5392–6.CrossRefGoogle Scholar
  15. 15.
    Chen G, Chen T. SPE speciation of inorganic arsenic in rice followed by hydride-generation atomic fluorescence spectrometric quantification. Talanta. 2014;119:202–6.CrossRefPubMedGoogle Scholar
  16. 16.
    Rasmussen RR, Qian Y, Sloth JJ. SPE HG-AAS method for the determination of inorganic arsenic in rice—results from method validation studies and a survey on rice products. Anal Bioanal Chem. 2013;405:7851–7.CrossRefPubMedGoogle Scholar
  17. 17.
    Welna M, Szymczycha-Madeja A, Pohl P. Comparison of strategies for sample preparation prior to spectrometric measurements for determination and speciation of arsenic in rice. Trends Anal Chem. 2015;65:122–36.CrossRefGoogle Scholar
  18. 18.
    Linquist BA, Anders MM, Adviento-Borbe MA, Chaney RL, Nalley LL, da Rosa EF, et al. Reducing greenhouse gas emissions, water use and grain arsenic levels in rice systems. Glob Chang Biol. 2015;21:407–17.CrossRefPubMedGoogle Scholar
  19. 19.
    Horwitz W, Albert R. The Horwitz ratio (HorRat): a useful index of method performance with respect to precision. J AOAC Int. 2006;89:1095–109.PubMedGoogle 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

  • Rufus L. Chaney
    • 1
    Email author
  • Carrie E. Green
    • 1
  • Steven J. Lehotay
    • 2
  1. 1.Adaptive Cropping Systems LaboratoryUSDA-Agricultural Research ServiceBeltsvilleUSA
  2. 2.Eastern Regional Research CenterUSDA Agricultural Research ServiceWyndmoorUSA

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