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

• Main Authors: Hongmin Jia, Tianjiao Zhou, Hong Zhu, Li Shen, Pingli He
• Format: Article
• Language: English
• Published: MDPI AG 2018-12-01
• Series: Molecules
• Subjects: soybean; Gly m 5.0101; β-conglycinin; protein quantification; liquid chromatography-tandem mass spectrometry
• Online Access: http://www.mdpi.com/1420-3049/24/1/68
• ISSN: 1420-3049

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.