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HALBMAYR-JECH ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 98, N O. 1, 2015 103
Gluten in Rice Flour and Baked Rice Products
by G12 Sandwich ELISA: First Action 2014.03
ELISABETH HALBMAYR-JECH
Romer Labs Division Holding GmbH, Technopark 1, 3430 Tulln, Austria
ADRIAN ROGERS
Romer Labs UK Ltd, Block 5, The Heath Technical and Business Park, Runcorn, Cheshire WA7 4QX, United Kingdom
CLYDE DON
Foodphysica, Vogelwikke 12, 6665 HP Driel, The Netherlands
MICHAEL PRINSTER
Romer Labs Inc, 1301 Stylemaster Dr, Union, MO 63084-1156
Collaborators: G. Augustin; C. Brewe; Z. Bugyi; D. Clarke; P. Cressey; A. Firzinger; J. Gelroth; M. Hemingway; R. Hochegger;
J. Jolly; P. Kasturi; P. Koehler; T. Koerner; M. Marquard; C. Poirier; A. Rogers; G. Sharma; R. Sherlock; C. Sousa; S. Taylor;
S. Tomoszi, J. Topping; P. Wehling
Received August 5, 2014.
The method was approved by the Expert Review Panel for Food
Allergens-Gluten as First Action.
The Expert Review Panel for Food Allergens-Gluten invites method
users to provide feedback on the First Action methods. Feedback from
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and are critical to gaining global recognition and acceptance of the
methods. Comments can be sent directly to the corresponding author
or
Corresponding author’s e-mail:
Copyright permission has been received by Cereal Foods World to
reprint the tables.
DOI:10.5740/jaoacint.14-197
FOOD COMPOSITION AND ADDITIVES
The Protein and Enzymes Technical Committee of
American Association of Cereal Chemists initiated
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has been validated for testing foods to conform
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AgraQuant® Gluten G12 is a sandwich ELISA for
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cross-bred varieties in various foodstuffs. The G12
antibody utilized in the test kit binds to the celiac toxic amino
acid sequence QPQLPY and related sequences in rye and
barley (1,2). A homogenized sample is extracted with ethanol
and a proprietary extraction solution containing reducing agents.
The gluten determination is based on a microtiter plate coated
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a peroxidase-labeled G12 antibody. The determination can be
done in 60 min. Ready-to-use standards of the ELISA test kit
are calibrated against the Working Group on Prolamin Analysis
and Toxicity (WGPAT) gliadin standard material and cover a
range from 4 to 200 mg gluten/kg sample (see Figure 1). The
preparation of ready-to-use standards was described at Halbmayr-
Jech et al. (3).
Single-laboratory validation (SLV), performed by Romer Labs
UK Ltd in May 2011, determined an LOD of 2 mg gluten/kg
sample and an LOQ of 4 mg gluten/kg sample (see Table 1) as
well as a recovery rate ranging from 90 to 145% (see Table 2) for
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for repeatability and lot-to-lot variation (reproducibility) was
15% or less determined within the SLV (see Tables 3–5). The
AgraQuant Gluten G12 kit furthermore produced results similar
to those assigned values for the current Codex type I approved
R5 Mendez method in three Food Analysis Performance
Assessment Scheme (FAPAS) rounds in 2011 (see Table 6).
The Gluten G12 Sandwich ELISA assay has been evaluated in
a collaborative study with 20 participants. The main target for an
allowable immunogenic gluten method according to the Codex
Alimentarius is that it should have a detection limit of 10 mg/kg
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study and discusses the results in relation to current thresholds
(20 mg/kg) for gluten-free products.
Collaborative Study
Study Design
The study was conducted on 12 different food samples
prepared in the laboratory of the Deutsche Forschungsanstalt für
Lebensmittelchemie, Freising, Germany. Blind-coded samples
in duplicate, ELISA test kits including extraction solution,
method instructions, and result reporting sheets were sent to all
participating laboratories.
Collaborators
The collaborative study was coordinated by Clyde Don,
Foodphysica, Driel, The Netherlands. Twenty laboratories from
the food producing industry, universities, governments, contract