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Proposal for C-Hordein as Reference Material in Gluten Quantification.

Xin Huang1, Päivi Kanerva2, Hannu Salovaara1

  • 1Department of Food and Environmental Sciences, University of Helsinki , Agnes Sjöberginkatu 2, PL 66, Helsinki FI-00014, Finland.

Journal of Agricultural and Food Chemistry
|February 25, 2017
PubMed
Summary

The R5 ELISA method overestimates barley prolamin (hordein) in gluten-free foods due to using a wheat gliadin standard. A C-hordein standard is proposed for accurate barley prolamin quantification.

Keywords:
C-hordeinR5gluten quantificationgluten-freehordeinprolamin

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Area of Science:

  • Food Science
  • Analytical Chemistry
  • Allergen Detection

Background:

  • Gluten-free products often contain barley prolamin (hordein), a potential allergen.
  • The R5 ELISA method is commonly used to quantify hordein but may overestimate its concentration when calibrated with wheat gliadin.

Purpose of the Study:

  • To investigate the reasons for overestimation of barley prolamin by R5 ELISA using a wheat gliadin standard.
  • To develop a more accurate method for quantifying barley prolamin in gluten-free products.

Main Methods:

  • Comparative analysis of R5 antibody reactivity towards hordein and gliadin.
  • Determination of C-hordein proportion in Finnish barley cultivars.
  • Development of a preparative-scale method for C-hordein isolation and purification.
  • Spiking experiments using gluten-free oat flour with barley flour.

Main Results:

  • The R5 antibody primarily recognizes C-hordein within whole hordein.
  • The proportion of C-hordein in Finnish barley cultivars (16-33%) is higher than its homologous protein, ω-gliadin.
  • Using a gliadin standard resulted in 1.8-2.5 times overestimation of barley prolamin in spiked samples.
  • A mixed standard of 40% C-hordein and inert protein yielded accurate estimates.

Conclusions:

  • The use of a wheat gliadin standard leads to significant overestimation of barley prolamin in gluten-free products.
  • C-hordein is the main reactive component of hordein for the R5 antibody.
  • Purified C-hordein is proposed as a suitable reference material for accurate barley prolamin quantification using R5 ELISA.