Quantification of genetically modified soya using strong anion exchange chromatography and time-of-flight mass spectrometry |
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Authors: | Po-Chih Chang P. Muralidhar Reddy Yen-Peng Ho |
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Affiliation: | 1. Department of Chemistry, National Dong Hwa University, Hualien, 97401, Taiwan, Republic of China 2. Department of Chemistry, Nizam College, Osmania University, Hyderabad, 500001, Andhra Pradesh, India
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Abstract: | Stable-isotope dimethyl labeling was applied to the quantification of genetically modified (GM) soya. The herbicide-resistant gene-related protein 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) was labeled using a dimethyl labeling reagent, formaldehyde-H2 or -D2. The identification and quantification of CP4 EPSPS was performed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The CP4 EPSPS protein was separated from high abundance proteins using strong anion exchange chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Then, the tryptic peptides from the samples and reference were labeled with formaldehyde-H2 and formaldehyde-D2, respectively. The two labeled pools were mixed and analyzed using MALDI-MS. The data showed a good correlation between the peak ratio of the H- and D-labeled peptides and the GM soya percentages at 0.5, 1, 3, and 5 %, with R 2 of 0.99. The labeling reagents are readily available. The labeling experiments and the detection procedures are simple. The approach is useful for the quantification of GM soya at a level as low as 0.5 %. |
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