Nagai Hiroyuki
Gifu Prefectural Research Institute for Health and Environmental Science, 1-1 Naka-Fudougaoka, Kakamigahara, 504-0838, Japan.
J AOAC Int. 2017 Jul 1;100(4):1051-1057. doi: 10.5740/jaoacint.11-0214.
An analysis technique using LC with tandem MS (MS/MS) has been developed for the determination of buckwheat proteins, including major allergens. A protein solution extracted from buckwheat was reduced, alkylated, and digested by trypsin. Peptide spectra were obtained using full-scan LC-MS/MS analysis, and peptide sequences were determined through a protein search. Nine peptides of the 13S globulin seed storage protein and one peptide of a 16 kDa allergen were selected as the marker peptides, and multiple reaction monitoring conditions were optimized. Using the conditions, different kinds of buckwheat noodles, powders, and other food ingredients were analyzed. As a result, buckwheat samples present all the fragment peaks, whereas other foods, including Sesamum indicum, wheat, and soybeans, are not detected at all. These findings indicate that LC-MS/MS analysis may be applied to the detection of buckwheat food allergens.
已开发出一种使用液相色谱串联质谱（MS/MS）的分析技术来测定荞麦蛋白，包括主要过敏原。从荞麦中提取的蛋白质溶液经还原、烷基化处理后，用胰蛋白酶消化。通过全扫描液相色谱-质谱/质谱分析获得肽谱，并通过蛋白质搜索确定肽序列。选择了13S球蛋白种子贮藏蛋白的9个肽段和一种16 kDa过敏原的1个肽段作为标记肽，并优化了多反应监测条件。使用这些条件，对不同种类的荞麦面条、粉末和其他食品成分进行了分析。结果，荞麦样品呈现出所有的碎片峰，而其他食品，包括芝麻、小麦和大豆，根本未被检测到。这些发现表明，液相色谱-质谱/质谱分析可应用于荞麦食品过敏原的检测。
