文章基本信息

标题：Comprehensive Characterization and Relative Quantification of α-Amylase/Trypsin Inhibitors from Wheat Cultivars by Targeted HPLC-MS/MS
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作者：Sorel Tchewonpi Sagu ; Lynn Zimmermann ; Eva Landgräber 等
期刊名称：Foods
电子版ISSN：2304-8158
出版年度：2020
卷号：9
期号：10
页码：1448-1472
DOI：10.3390/foods9101448
出版社：MDPI Publishing
摘要：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.
关键词：α-amylase/trypsin inhibitors; wheat cultivars; SDS-PAGE; peptides markers; relative quantification; mass spectrometry; LC-MRM-MS α-amylase/trypsin inhibitors ; wheat cultivars ; SDS-PAGE ; peptides markers ; relative quantification ; mass spectrometry ; LC-MRM-MS