Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS Proteomics
The increasing popularity of “superseeds” such as flax, sesame, amaranth and quinoa as functional foods raises the need for robust analytical methods for authentication purposes. In this work, a standardized workflow for the extraction, characterization and identification of unique peptides that may...
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MDPI AG
2025-07-01
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| author | Sorel Tchewonpi Sagu Beatrice Schnepf Peter Stenzel Kapil Nichani Alexander Erban Joachim Kopka Harshadrai M. Rawel Andrea Henze |
| author_facet | Sorel Tchewonpi Sagu Beatrice Schnepf Peter Stenzel Kapil Nichani Alexander Erban Joachim Kopka Harshadrai M. Rawel Andrea Henze |
| author_sort | Sorel Tchewonpi Sagu |
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| description | The increasing popularity of “superseeds” such as flax, sesame, amaranth and quinoa as functional foods raises the need for robust analytical methods for authentication purposes. In this work, a standardized workflow for the extraction, characterization and identification of unique peptides that may be used as markers to distinguish superseed species was investigated. Ammonium bicarbonate/urea (Ambi/urea) extraction, sodium dodecyl sulfate (SDS) buffer and trichloroacetic acid (TCA) precipitation were initially implemented and, based on the level and composition of the extracted proteins, the SDS buffer protocol was selected. Electrophoresis analysis revealed consistent protein profiles between biological replicates from each of the eleven seed species, confirming the reproducibility of the SDS buffer protocol. Targeted mass spectrometry successfully identified species-specific peptide markers for six of eleven superseeds investigated, including peptides from conlinins in flaxseed (WVQQAK), 11S globulins in sesame (LVYIER), oleosin in quinoa (DVGQTIESK), agglutin-like lectins in amaranth (CAGVSVIR), as well as cupin-like proteins in poppy seeds (INIVNSQK) and edestins in hemp seeds (FLQLSAER). Moreover, proteome cross-analysis allowed us to disqualify the isomeric peptide LTALEPTNR from 11S globulins present in amaranth and quinoa. However, no reliable markers were identified for chia, canihua, basil, black cumin, and psyllium seeds under current conditions. While this targeted proteomics approach shows promise for superseed authentication, comprehensive method validation and alternative strategies for marker-deficient species are required before routine implementation. |
| format | Article |
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| institution | Kabale University |
| issn | 1420-3049 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | MDPI AG |
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| series | Molecules |
| spelling | doaj-art-bbed5141e636430d96cde836dbdd10122025-08-20T03:36:22ZengMDPI AGMolecules1420-30492025-07-013014299310.3390/molecules30142993Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS ProteomicsSorel Tchewonpi Sagu0Beatrice Schnepf1Peter Stenzel2Kapil Nichani3Alexander Erban4Joachim Kopka5Harshadrai M. Rawel6Andrea Henze7Institute of Agricultural and Nutritional Sciences, Martin Luther University Halle-Wittenberg, Von-Danckelmann-Platz 2, 06120 Halle (Saale), GermanyInstitute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, GermanyMax-Planck-Institut of Molecular Plant Physiology, Potsdam Science Park, Am Mühlenberg 1, 14476 Potsdam, GermanyInstitute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, GermanyMax-Planck-Institut of Molecular Plant Physiology, Potsdam Science Park, Am Mühlenberg 1, 14476 Potsdam, GermanyMax-Planck-Institut of Molecular Plant Physiology, Potsdam Science Park, Am Mühlenberg 1, 14476 Potsdam, GermanyInstitute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, GermanyInstitute of Agricultural and Nutritional Sciences, Martin Luther University Halle-Wittenberg, Von-Danckelmann-Platz 2, 06120 Halle (Saale), GermanyThe increasing popularity of “superseeds” such as flax, sesame, amaranth and quinoa as functional foods raises the need for robust analytical methods for authentication purposes. In this work, a standardized workflow for the extraction, characterization and identification of unique peptides that may be used as markers to distinguish superseed species was investigated. Ammonium bicarbonate/urea (Ambi/urea) extraction, sodium dodecyl sulfate (SDS) buffer and trichloroacetic acid (TCA) precipitation were initially implemented and, based on the level and composition of the extracted proteins, the SDS buffer protocol was selected. Electrophoresis analysis revealed consistent protein profiles between biological replicates from each of the eleven seed species, confirming the reproducibility of the SDS buffer protocol. Targeted mass spectrometry successfully identified species-specific peptide markers for six of eleven superseeds investigated, including peptides from conlinins in flaxseed (WVQQAK), 11S globulins in sesame (LVYIER), oleosin in quinoa (DVGQTIESK), agglutin-like lectins in amaranth (CAGVSVIR), as well as cupin-like proteins in poppy seeds (INIVNSQK) and edestins in hemp seeds (FLQLSAER). Moreover, proteome cross-analysis allowed us to disqualify the isomeric peptide LTALEPTNR from 11S globulins present in amaranth and quinoa. However, no reliable markers were identified for chia, canihua, basil, black cumin, and psyllium seeds under current conditions. While this targeted proteomics approach shows promise for superseed authentication, comprehensive method validation and alternative strategies for marker-deficient species are required before routine implementation.https://www.mdpi.com/1420-3049/30/14/2993superfoodssuperseedsauthenticationprotein extractionmarker peptidetargeted mass spectrometry |
| spellingShingle | Sorel Tchewonpi Sagu Beatrice Schnepf Peter Stenzel Kapil Nichani Alexander Erban Joachim Kopka Harshadrai M. Rawel Andrea Henze Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS Proteomics Molecules superfoods superseeds authentication protein extraction marker peptide targeted mass spectrometry |
| title | Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS Proteomics |
| title_full | Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS Proteomics |
| title_fullStr | Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS Proteomics |
| title_full_unstemmed | Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS Proteomics |
| title_short | Discovery of Species-Specific Peptide Markers for Superseed Authentication Using Targeted LC-MS/MS Proteomics |
| title_sort | discovery of species specific peptide markers for superseed authentication using targeted lc ms ms proteomics |
| topic | superfoods superseeds authentication protein extraction marker peptide targeted mass spectrometry |
| url | https://www.mdpi.com/1420-3049/30/14/2993 |
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