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Metabolomic Strategies Based on High-Resolution Mass Spectrometry as a Tool for Recognition of GMO (MON 89788 Variety) and Non-GMO Soybean: a Critical Assessment of Two Complementary Methods

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Abstract

This study was focused on distinguishing of genetically modified organism (Monsanto 89788 variety) and conventional soybeans by employing high-resolution mass spectrometry (HRMS) techniques for non-target screening of sample extracts. Two hyphenated instrumental platforms represented by (i) ultrahigh-performance liquid chromatography (U-HPLC) coupled to quadrupole/time of flight and (ii) ambient mass spectrometry with direct analysis in real time (DART) ion source-coupled OrbitrapMS were used. The statistical processing of generated data (metabolomic fingerprints) was performed by multivariate data analysis; principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) showed that both employed techniques enabled correct classification of genetically modified and conventional soybeans. In addition, some phosphatidylcholines and sugars were identified as the most significant markers.

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Authors and Affiliations

  1. Faculty of Food and Biochemical Technology, Department of Food Analysis and Nutrition, University of Chemistry and Technology, Prague, Technicka 3, 166 28, Prague 6, Czech Republic

    Vojtech Hrbek, Veronika Krtkova, Josep Rubert, Hana Chmelarova & Jana Hajslova

  2. Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technicka 3, 166 28, Prague 6, Czech Republic

    Katerina Demnerova

  3. Crop Research Institute, Prague, Drnovska 507/73, 161 06, Prague 6, Czech Republic

    Jaroslava Ovesna

Authors
  1. Vojtech Hrbek
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  2. Veronika Krtkova
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  3. Josep Rubert
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  4. Hana Chmelarova
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  5. Katerina Demnerova
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  6. Jaroslava Ovesna
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  7. Jana Hajslova
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Corresponding author

Correspondence to Jana Hajslova.

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Funding

This work was supported by the Ministry of Agriculture of the Czech Republic (NAZV-QI101B267), Operational Programme Prague—Competitiveness (CZ.2.16/3.1.00/21537 and CZ.2.16/3.1.00/24503) and by the ‘National Program of Sustainability I’—NPU I (LO1601—No. MSMT-43760/2015). Josep Rubert thanks the Generalitat Valenciana (Conselleria d’Educació, Cultura i Esport) for the VALi + d postdoctoral fellowship ‘Contractació de personal investigador en formació en fase postdoctoral 2014’ (APOSTD/2014/120).

Conflict of Interest

Vojtech Hrbek declares that he/she has no conflict of interest. Veronika Krtkova declares that he/she has no conflict of interest. Josep Rubert declares that he/she has no conflict of interest. Hana Chmelarova declares that he/she has no conflict of interest. Katerina Demnerova declares that he/she has no conflict of interest. Jaroslava Ovesna declares that he/she has no conflict of interest. Jana Hajslova declares that he/she has no conflict of interest.

Compliance with Ethical Requirements

This article does not contain any studies with human participants or animals performed by any of the authors.

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Electronic Supplementary Material

Figure S1

Examples of U-HPLC-ESI (+)-QTOFMS chromatographic records for free and bound forms of phytoestrogens present in soybean samples - 80% aqueous methanolic extract of conventional soybean sample. (TIFF 331 kb)

High Resolution Image (GIF 195 kb)

Figure S2

Permutation plots for the data obtained by U-HPLC-HRMS analysis of GMO and non-GMO soybean samples in both ionization modes. (TIFF 713 kb)

High Resolution Image (GIF 283 kb)

Figure S3

Permutation plots for the data obtained by DART-HRMS analysis of GMO and non-GMO soybean samples in both ionization modes. (TIFF 790 kb)

High Resolution Image (GIF 277 kb)

Figure S4

Chemometric analysis of data generated by LC-MS and DART-MS for conventional (grey), GMO (black) soybean samples and quality control (QC) samples (red triangles), PCA in positive and negative mode. (TIFF 992 kb)

High Resolution Image (GIF 350 kb)

Figure S5

S-plot of features (LC-ESI-HRMS analysis in positive mode) - ´markers´ with the highest importance for classification are highlighted in the black ovals. (TIFF 267 kb)

High Resolution Image (GIF 155 kb)

Figure S6

VIP plots for data obtained by both techniques in both ionization modes. (TIFF 356 kb)

High Resolution Image (GIF 121 kb)

Figure S7

Trend plots for data obtained by both techniques (LC-MS and DART-MS) in both ionization modes for conventional (grey) and GMO (black) soybean samples. (TIFF 1289 kb)

High Resolution Image (GIF 381 kb)

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Hrbek, V., Krtkova, V., Rubert, J. et al. Metabolomic Strategies Based on High-Resolution Mass Spectrometry as a Tool for Recognition of GMO (MON 89788 Variety) and Non-GMO Soybean: a Critical Assessment of Two Complementary Methods. Food Anal. Methods 10, 3723–3737 (2017). https://doi.org/10.1007/s12161-017-0929-8

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