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Gel-Based and Gel-Free Analytical Methods for the Detection of HMW-GS and LMW-GS in Wheat Flour

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Abstract

Durum wheat (Triticum turgidum L.) flour is instrumental for the production of pasta worldwide. The quality of this food rests on flour processing and on its protein content and composition. Gluten proteins as high and low-molecular weight glutenins (GS) are important to predict the flour technological property in pasta making. Different methods were compared to separate, identify and quantify GS in flours from two wheat cultivars. Sodium dodecyl sulphate-polyacrilamide gel electrophoresis (SDS-PAGE) gave in a fast way information about the GS assets. Two-dimensional gel electrophoresis (2D-GE) allowed for the highest resolution in detecting and quantifying single GS, subsequently identified by liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS/MS). Reversed-phase high-performance liquid chromatography (RP-HPLC) is a non-gel alternative system for separation and quantification of single GS that when combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) gave information about their exact masses. This method gives also quantitative indications of each individual GS. Different GS patterns and contents were detected in the flour of the two cultivars, underlining the importance of these analytical methods before determining the best flour processing procedure in pasta making. The different methods were evaluated with a modular approach consisting of a grid of different parameters and a non-linear score within each module.

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Abbreviations

GS:

Glutenins

SDS-PAGE:

Sodium dodecyl sulphate-polyacrilamide gel electrophoresis

2D-GE:

Two-dimensional gel electrophoresis

LC-ESI-MS/MS:

Liquid chromatography-electrospray ionization mass spectrometry

RP-HPLC:

Reversed-phase high-performance liquid chromatography

MALDI-TOF/MS:

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

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Acknowledgments

The authors thank the University of Parma, Regione Elimia-Romagna (IT) SITEA.PARMA Technopole (POR FESR 2007–2013) (MN) and the Interdepartmental Measure Centre “Giuseppe Casnati” (CIM) for accessing to MS facilities. The contribution of COST action FA0905, coordinated by Prof. Bal Ram Singh (Norwegian University of Life sciences, Norway), is also acknowledged.

Funding

This work was funded by AGER grant 2010–0278 “Environmental and economic sustainability for yield and quality production of durum wheat supply chain”.

Conflict of Interest

Dr. Giovanna Visioli declares that she has no conflict of interest. Dr. Alessia Comastri declares that she has no conflict of interest. Dr. Davide Imperiale declares that he has no conflict of interest. Dr. Gianluca Paredi declares that he has no conflict of interest. Dr. Andrea Faccini declares that he has no conflict of interest. Prof. Nelson Marmiroli declares that he has no conflict of interest. This article does not contain any studies with human or living animal subjects.

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Authors and Affiliations

  1. Department of Life Sciences, University of Parma, Parco Area delle Scienze 11/a, 43124, Parma, Italy

    Giovanna Visioli, Alessia Comastri, Davide Imperiale & Nelson Marmiroli

  2. Department of Pharmacy, Center Siteia.Parma, University of Parma, Parco Area delle Scienze 23/a, 43124, Parma, Italy

    Gianluca Paredi

  3. Interdepartmental Measure Centre “Giuseppe Casnati”, University of Parma, Parco Area delle Scienze 24/a, 43124, Parma, Italy

    Andrea Faccini

Authors
  1. Giovanna Visioli
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  2. Alessia Comastri
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  3. Davide Imperiale
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  4. Gianluca Paredi
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  5. Andrea Faccini
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  6. Nelson Marmiroli
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Corresponding author

Correspondence to Giovanna Visioli.

Electronic supplementary material

In Supplementary Material, a Table reporting agronomical characteristics of the two cultivars and two figures reporting mass spectra of two peptide sequences were given.

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Visioli, G., Comastri, A., Imperiale, D. et al. Gel-Based and Gel-Free Analytical Methods for the Detection of HMW-GS and LMW-GS in Wheat Flour. Food Anal. Methods 9, 469–476 (2016). https://doi.org/10.1007/s12161-015-0218-3

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  • DOI: https://doi.org/10.1007/s12161-015-0218-3

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