TY  - JOUR
A1  - Sagu Tchewonpi, Sorel
A1  - Zimmermann, Lynn
A1  - Landgräber, Eva
A1  - Homann, Thomas
A1  - Huschek, Gerd
A1  - Özpinar, Haydar
A1  - Schweigert, Florian J.
A1  - Rawel, Harshadrai Manilal
T1  - Comprehensive Characterization and Relative Quantification of α-Amylase/Trypsin Inhibitors from Wheat Cultivars by Targeted HPLC-MS/MS
JF  - Foods
N2  - 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.
KW  - α-amylase/trypsin inhibitors
KW  - wheat cultivars
KW  - SDS-PAGE
KW  - peptides markers
KW  - relative quantification
KW  - mass spectrometry
KW  - LC-MRM-MS
Y1  - 2020
U6  - https://doi.org/10.3390/foods9101448
SN  - 2304-8158
VL  - 9
IS  - 10
PB  - MDPI
CY  - Basel
ER  - 
TY  - GEN
A1  - Sagu Tchewonpi, Sorel
A1  - Zimmermann, Lynn
A1  - Landgräber, Eva
A1  - Homann, Thomas
A1  - Huschek, Gerd
A1  - Özpinar, Haydar
A1  - Schweigert, Florian J.
A1  - Rawel, Harshadrai Manilal
T1  - Comprehensive Characterization and Relative Quantification of α-Amylase/Trypsin Inhibitors from Wheat Cultivars by Targeted HPLC-MS/MS
T2  - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe
N2  - 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.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 1028 
KW  - α-amylase/trypsin inhibitors
KW  - wheat cultivars
KW  - SDS-PAGE
KW  - peptides markers
KW  - relative quantification
KW  - mass spectrometry
KW  - LC-MRM-MS
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-486118
SN  - 1866-8372
IS  - 1028
ER  -