Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled Experiments
Analysis of plant metabolite <sup>13</sup>C-enrichments with gas-chromatography mass spectrometry (GC/MS) has gained interest recently. By combining multiple fragments of a trimethylsilyl (TMS) derivative, <sup>13</sup>C-positional enrichments can be calculated. However, this...
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MDPI AG
2023-03-01
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| Series: | Metabolites |
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| Online Access: | https://www.mdpi.com/2218-1989/13/4/466 |
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| author | Younès Dellero Olivier Filangi Alain Bouchereau |
| author_facet | Younès Dellero Olivier Filangi Alain Bouchereau |
| author_sort | Younès Dellero |
| collection | DOAJ |
| description | Analysis of plant metabolite <sup>13</sup>C-enrichments with gas-chromatography mass spectrometry (GC/MS) has gained interest recently. By combining multiple fragments of a trimethylsilyl (TMS) derivative, <sup>13</sup>C-positional enrichments can be calculated. However, this new approach may suffer from analytical biases depending on the fragments selected for calculation leading to significant errors in the final results. The goal of this study was to provide a framework for the validation of <sup>13</sup>C-positional approaches and their application to plants based on some key metabolites (glycine, serine, glutamate, proline, α-alanine and malate). For this purpose, we used tailor-made <sup>13</sup>C-PT standards, harboring known carbon isotopologue distributions and <sup>13</sup>C-positional enrichments, to evaluate the reliability of GC-MS measurements and positional calculations. Overall, we showed that some mass fragments of proline_2TMS, glutamate_3TMS, malate_3TMS and α-alanine_2TMS had important biases for <sup>13</sup>C measurements resulting in significant errors in the computational estimation of <sup>13</sup>C-positional enrichments. Nevertheless, we validated a GC/MS-based <sup>13</sup>C-positional approach for the following atomic positions: (i) C1 and C2 of glycine_3TMS, (ii) C1, C2 and C3 of serine_3TMS, and (iii) C1 of malate_3TMS and glutamate_3TMS. We successfully applied this approach to plant <sup>13</sup>C-labeled experiments for investigating key metabolic fluxes of plant primary metabolism (photorespiration, tricarboxylic acid cycle and phosphoenolpyruvate carboxylase activity). |
| first_indexed | 2024-03-11T04:45:36Z |
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| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-11T04:45:36Z |
| publishDate | 2023-03-01 |
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| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-59f7bcd9de584fa29c811ae458ad6cba2023-11-17T20:23:52ZengMDPI AGMetabolites2218-19892023-03-0113446610.3390/metabo13040466Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled ExperimentsYounès Dellero0Olivier Filangi1Alain Bouchereau2Institute for Genetics, Environment and Plant Protection (IGEPP), National Research Institute for Agriculture, Food and Environment (INRAE), Institut Agro, Université Rennes, 35650 Le Rheu, FranceInstitute for Genetics, Environment and Plant Protection (IGEPP), National Research Institute for Agriculture, Food and Environment (INRAE), Institut Agro, Université Rennes, 35650 Le Rheu, FranceInstitute for Genetics, Environment and Plant Protection (IGEPP), National Research Institute for Agriculture, Food and Environment (INRAE), Institut Agro, Université Rennes, 35650 Le Rheu, FranceAnalysis of plant metabolite <sup>13</sup>C-enrichments with gas-chromatography mass spectrometry (GC/MS) has gained interest recently. By combining multiple fragments of a trimethylsilyl (TMS) derivative, <sup>13</sup>C-positional enrichments can be calculated. However, this new approach may suffer from analytical biases depending on the fragments selected for calculation leading to significant errors in the final results. The goal of this study was to provide a framework for the validation of <sup>13</sup>C-positional approaches and their application to plants based on some key metabolites (glycine, serine, glutamate, proline, α-alanine and malate). For this purpose, we used tailor-made <sup>13</sup>C-PT standards, harboring known carbon isotopologue distributions and <sup>13</sup>C-positional enrichments, to evaluate the reliability of GC-MS measurements and positional calculations. Overall, we showed that some mass fragments of proline_2TMS, glutamate_3TMS, malate_3TMS and α-alanine_2TMS had important biases for <sup>13</sup>C measurements resulting in significant errors in the computational estimation of <sup>13</sup>C-positional enrichments. Nevertheless, we validated a GC/MS-based <sup>13</sup>C-positional approach for the following atomic positions: (i) C1 and C2 of glycine_3TMS, (ii) C1, C2 and C3 of serine_3TMS, and (iii) C1 of malate_3TMS and glutamate_3TMS. We successfully applied this approach to plant <sup>13</sup>C-labeled experiments for investigating key metabolic fluxes of plant primary metabolism (photorespiration, tricarboxylic acid cycle and phosphoenolpyruvate carboxylase activity).https://www.mdpi.com/2218-1989/13/4/466mass spectrometrygas chromatographyisotopebrassica napusphotorespirationtricarboxylic acid cycle |
| spellingShingle | Younès Dellero Olivier Filangi Alain Bouchereau Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled Experiments Metabolites mass spectrometry gas chromatography isotope brassica napus photorespiration tricarboxylic acid cycle |
| title | Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled Experiments |
| title_full | Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled Experiments |
| title_fullStr | Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled Experiments |
| title_full_unstemmed | Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled Experiments |
| title_short | Evaluation of GC/MS-Based <sup>13</sup>C-Positional Approaches for TMS Derivatives of Organic and Amino Acids and Application to Plant <sup>13</sup>C-Labeled Experiments |
| title_sort | evaluation of gc ms based sup 13 sup c positional approaches for tms derivatives of organic and amino acids and application to plant sup 13 sup c labeled experiments |
| topic | mass spectrometry gas chromatography isotope brassica napus photorespiration tricarboxylic acid cycle |
| url | https://www.mdpi.com/2218-1989/13/4/466 |
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