Filtern
Volltext vorhanden
- nein (1)
Erscheinungsjahr
- 2020 (1)
Dokumenttyp
- Wissenschaftlicher Artikel (1) (entfernen)
Sprache
- Englisch (1)
Gehört zur Bibliographie
- ja (1)
Schlagworte
- LC-MRM-MS (1)
- SDS-PAGE (1)
- mass spectrometry (1)
- peptides markers (1)
- relative quantification (1)
- wheat cultivars (1)
- α-amylase/trypsin inhibitors (1)
Institut
- Institut für Ernährungswissenschaft (1) (entfernen)
The α-amylase/trypsin inhibitors (ATIs) are discussed as being responsible for non-celiac wheat sensitivity (NCWS), besides being known as allergenic components for baker’s asthma. Different approaches for characterization and quantification including proteomics-based methods for wheat ATIs have been documented. In these studies generally the major ATIs have been addressed. The challenge of current study was then to develop a more comprehensive workflow encompassing all reviewed wheat-ATI entries in UniProt database. To substantially test proof of concept, 46 German and Turkish wheat samples were used. Two extractions systems based on chloroform/methanol mixture (CM) and under buffered denaturing conditions were evaluated. Three aspects were optimized, tryptic digestion, chromatographic separation, and targeted tandem mass spectrometric analysis (HPLC-MS/MS). Preliminary characterization with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) documented the purity of the extracted ATIs with CM mixture and the amylase (60–80%)/trypsin (10–20%) inhibition demonstrated the bifunctional activity of ATIs. Thirteen (individual/common) biomarkers were established. Major ATIs (7–34%) were differently represented in samples. Finally, to our knowledge, the proposed HPLC-MS/MS method allowed for the first time so far the analysis of all 14 reviewed wheat ATI entries reported.