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Rye secalin isolates to develop reference materials for gluten detection.

Majlinda Xhaferaj¹, Gabriella Muskovics², Zsuzsanna Bugyi³

¹Karlsruhe Institute of Technology, Institute of Applied Biosciences, Department of Bioactive and Functional Food Chemistry, Karlsruhe, Germany.

|January 11, 2025

Summary

Developing accurate gluten testing for celiac disease patients is crucial. This study created rye gluten isolates as standards, improving gluten analysis in gluten-free products contaminated with rye.

Keywords:

Celiac disease Enzyme-linked immunosorbent assay (ELISA)Liquid chromatography–tandem mass spectrometry (LC-MS/MS)Reversed-phase high-performance liquid chromatography (RP-HPLC)Secalins Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE)

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Area of Science:

Food science
Analytical chemistry
Celiac disease research

Background:

Gluten-free products require <20 mg/kg gluten for celiac safety.
Current analytical methods lack rye-specific reference materials.
Rye contamination in gluten-free foods poses an analytical challenge.

Purpose of the Study:

To investigate the impact of harvest year on rye gluten composition.
To develop distinct rye protein isolates for use as calibration standards.
To enhance the accuracy of gluten testing in rye-containing foods.

Main Methods:

Extraction of rye proteins using four different procedures: prolamins (PROL), glutelins (GLUT), gluten (G), and acetonitrile/water-extractable proteins (AWEP).
Characterization of isolates via RP-HPLC, GP-HPLC, SDS-PAGE, and LC-MS/MS.
Evaluation of isolates using R5 ELISA to determine their response order.

Main Results:

Four distinct rye protein isolates were successfully generated.
Isolate characterization confirmed their unique compositions.
R5 ELISA showed the response order: PROLiso > AWEPiso > Giso > GLUTiso.

Conclusions:

This study provides essential rye-specific reference materials for gluten analysis.
The developed isolates are crucial for accurate quantification of gluten in rye-contaminated foods.
Findings represent a significant advancement in ensuring the safety of gluten-free products.