Plant and Soil

, Volume 361, Issue 1–2, pp 251–260 | Cite as

Energy-dispersive X-ray fluorescence analysis of zinc and iron concentration in rice and pearl millet grain

  • Nicholas G. Paltridge
  • Lachlan J. Palmer
  • Paul J. Milham
  • Georgia E. Guild
  • James C. R. Stangoulis
Regular Article


Background and aims

Rice (Oryza sativa L.) and pearl millet (Pennisetum glaucum L.) biofortification breeding programs require accurate and convenient methods to identify nutrient dense genotypes. The aim of this study was to investigate energy-dispersive X-ray fluorescence spectrometry (EDXRF) for the measurement of zinc (Zn) and iron (Fe) concentration in whole grain rice and pearl millet.


Grain samples were obtained from existing biofortification breeding programs. Reference Zn and Fe concentrations obtained by inductively-coupled plasma-optical emission spectroscopy (ICP-OES) were used to calibrate the EDXRF instrument. Calibration was performed with 24 samples and separate calibrations were developed for rice and pearl millet. To validate calibrations, EDXRF analyses were conducted on an additional 40 samples of each species.


EDXRF results were highly correlated with ICP-OES values for both Zn and Fe in both species (r2 = 0.79 to 0.98). EDXRF predicted Zn and Fe in rice to within 1.9 and 1.6 mg kg−1 of ICP-OES values, and Zn and Fe in pearl millet to within 7.6 and 12.5 mg kg−1 of ICP-OES values, at a 95% confidence level.


EDXRF offers a convenient, economical tool for screening Zn and Fe concentration in rice and pearl millet biofortification breeding programs.


XRF EDXRF Biofortification Micronutrient Plant 



Funding for this work was provided by HarvestPlus. We thank Oxford Instruments and Neal Robson for help identifying optimal Zn and Fe EDXRF conditions, Parminder Virk and Alamgir Hossain for supplying rice samples, Kedar Rai for supplying pearl millet samples, Waite Analytical Services for ICP-OES analysis, Zarina Yasmin for technical assistance, and Robin Graham and Wendy Telfer for helpful suggestions on the manuscript.


  1. Arai T (2006) Introduction. In: Beckhoff B, Kanngießer B, Langhoff N, Wedell R, Wolff H (eds) Handbook of practical X-ray fluorescence analysis. Springer, Berlin Heidelberg, pp 1–31CrossRefGoogle Scholar
  2. Blank AB, Eksperiandova LP (1998) Specimen preparation in X-ray fluorescence analysis of materials and natural objects. X-ray Spectrom 27:147–160CrossRefGoogle Scholar
  3. Choi EY, Graham R, Stangoulis J (2007) Semi-quantitative analysis for selecting Fe- and Zn-dense genotypes of staple food crops. J Food Compos Anal 20:496–505CrossRefGoogle Scholar
  4. Clark RB, Frank KD, Zaifnejad M, Denning J (1992) X-ray fluorescence analysis of small leaf samples mixed with cellulose or boric acid. Commun Soil Sci Plant Anal 23:569–583CrossRefGoogle Scholar
  5. Ekinci N, Ekinci R, Sahin Y (2002) Determination of iodine and calcium concentrations in the bread improver using ED-XRF. J Quant Spectrosc Radiat Transf 74:783–787CrossRefGoogle Scholar
  6. Frank KD, Burch J, Denning J (1992) Mineral analysis of corn leaves by X-ray fluorescence on ground versus unground leaf samples. Commun Soil Sci Plant Anal 23:2415–2424CrossRefGoogle Scholar
  7. Injuk J, Van Grieken R, Blank A, Eksperiandova L, Buhrke V (2006) Specimen preparation. In: Beckhoff B, Kanngießer B, Langhoff N, Wedell R, Wolff H (eds) Handbook of practical X-ray fluorescence analysis. Springer, Berlin Heidelberg, pp 411–432CrossRefGoogle Scholar
  8. Kocman V, Peel TE, Tomlinson GH (1991) Rapid analysis of macro and micro nutrients in leaves and vegetation by automated x-ray-fluorescence spectrometry (a case study of an acid-rain affected forest). Commun Soil Sci Plant Anal 22:2063–2075CrossRefGoogle Scholar
  9. Melquiades FL, Appolini CR (2004) Application of XRF for environmental analysis. J Radioanal Nucl Chem 262:533–541CrossRefGoogle Scholar
  10. Nestel P, Bouis HE, Meenakshi JV, Pfeiffer W (2006) Biofortification of staple food crops. J Nutr 136:1064–1067PubMedGoogle Scholar
  11. Noda T, Tsuda S, Mori M, Takigawa S, Matsuura-Endo C, Kim S-J, Hashimoto N, Yamauchi H (2006) Determination of the phosphorus content in potato starch using an energy-dispersive X-ray fluorescence method. Food Chem 95:632–637CrossRefGoogle Scholar
  12. Ozturk L, Yazici MA, Yucel C, Torun A, Cekic C, Bagci A, Ozkan H, Braun HJ, Sayers Z, Cakmak I (2006) Concentration and localization of zinc during seed development and germination in wheat. Physiol Plant 128:144–152CrossRefGoogle Scholar
  13. Perring L, Andrey D (2003) ED-XRF as a tool for rapid minerals control in milk-based products. J Agric Food Chem 51:4207–4212PubMedCrossRefGoogle Scholar
  14. Perring L, Andrey D, Basic-Dvorzak M, Hammer D (2005) Rapid quantification of iron, copper and zinc in food premixes using energy dispersive X-ray fluorescence. J Food Compos Anal 18:655–663Google Scholar
  15. Perring L, Blanc J (2007) EDXRF determination of iron during infant cereals production and its fitness for purpose. Int J Food Sci Technol 42:551–555CrossRefGoogle Scholar
  16. Perring L, Blanc J (2008) Validation of quick measurement of mineral nutrients in milk powders: comparison of energy dispersive X-ray fluorescence with inductively coupled plasma-optical emission spectroscopy and potentiometry reference methods. Sens Instrum Food Qual 2:254–261CrossRefGoogle Scholar
  17. Perring L, Monard F (2010) Improvement of Energy Dispersive X-Ray Fluorescence throughput: influence of measuring times and number of replicates on validation performance characteristics. Food Anal Methods 3:104–115CrossRefGoogle Scholar
  18. Pfeiffer WH, McClafferty B (2007) Biofortification: Breeding micronutrient-dense crops. In: Kang MS, Priyadarshan PM (eds) Breeding major food staples. Blackwell, Ames, pp 61–91CrossRefGoogle Scholar
  19. Prom-u-thai C, Dell B, Thomson G, Rerkssem B (2003) Easy and rapid detection of iron in rice seed. Sci Asia 29:314–317CrossRefGoogle Scholar
  20. Rousseau RM, Willis JP, Duncan AR (1996) Practical XRF calibration procedures for major and trace elements. X-ray Spectrom 25:179–189CrossRefGoogle Scholar
  21. Velu G, Ortiz-Monasterio I, Singh RP, Payne T (2011) Variation for grain micronutrients concentration in wheat core-collection accessions of diverse origin. Asian J Crop Sci 3:43–48CrossRefGoogle Scholar
  22. West M, Ellis AT, Potts PJ, Streli C, Vanhoof C, Wegrzynek D, Wobrauschek P (2009) Atomic spectrometry update. X-Ray fluorescence spectrometry. J Anal At Spectrom 24:1289–1326CrossRefGoogle Scholar
  23. West M, Ellis AT, Potts PJ, Streli C, Vanhoof C, Wegrzynek D, Wobrauschek P (2010) Atomic spectrometry update–X-Ray fluorescence spectrometry. J Anal At Spectrom 25:1503–1545CrossRefGoogle Scholar
  24. Wheal MS, Fowles TO, Palmer LT (2011) A cost-effective acid digestion method using closed polypropylene tubes for inductively coupled plasma optical emission spectrometry (ICP-OES) analysis of plant essential elements. Anal Methods (in press) doi: 10.1039/c1ay05430a
  25. Zarcinas BA, Cartwright B, Spouncer LR (1987) Nitric acid digestion and multi-element analysis of plant material by inductively coupled plasma spectrometry. Commun Soil Sci Plant Anal 18:131–146CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Nicholas G. Paltridge
    • 1
  • Lachlan J. Palmer
    • 1
  • Paul J. Milham
    • 2
  • Georgia E. Guild
    • 1
  • James C. R. Stangoulis
    • 1
  1. 1.School of Biological SciencesFlinders UniversityAdelaideAustralia
  2. 2.NSW Department of Primary IndustriesRichmondAustralia

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