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Quantification of Gly m 5.0101 in Soybean and Soy Products by Liquid Chromatography-Tandem Mass Spectrometry

機(jī)譯：液相色譜-串聯(lián)質(zhì)譜法定量測(cè)定大豆和豆制品中的Gly m 5.0101

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摘要

Gly m 5.0101, the alpha subunit of β-conglycinin, is one of the major allergens found in soybeans that has been identified as causing an allergic reaction. Here, we developed a quantification method of Gly m 5.0101 with multiple reaction monitoring using the synthetic peptide 194NPFLFGSNR202 as the external standard. Firstly, the ground soybean was defatted and extracted with a protein extraction buffer. Then the crude extract was on-filter digested by trypsin and analyzed by liquid chromatography-tandem mass spectrometry. The selected peptide exhibited a detection limit of 0.48 ng/mL and a linear relationship in a concentration range from 1.6 to 500 ng/mL (r2 > 0.99). The developed method was successfully applied to quantify the Gly m 5.0101 level in dozens of soybean varieties from different sources and soybean products derived from different processing techniques. The developed method could be used to further analyze β-conglycinin in soybean seeds combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis.
機(jī)譯：β-伴大豆球蛋白的α亞基Gly m 5.0101是大豆中發(fā)現(xiàn)的主要過(guò)敏原之一，已被確定會(huì)引起過(guò)敏反應(yīng)。在這里，我們開(kāi)發(fā)了一種以合成肽 194 NPFLFGSNR 202 作為外標(biāo)進(jìn)行多反應(yīng)監(jiān)測(cè)的Gly m 5.0101定量方法。首先，將磨碎的大豆脫脂并用蛋白質(zhì)提取緩沖液提取。然后將粗提取物在過(guò)濾器上用胰蛋白酶消化，并通過(guò)液相色譜-串聯(lián)質(zhì)譜法進(jìn)行分析。所選肽的檢測(cè)限為0.48 ng / mL，濃度范圍為1.6至500 ng / mL（r 2

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