Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation Technique
Lyophilization is a common method used for stabilizing biological samples prior to storage or to concentrate extracts. However, it is possible that this process may alter the metabolic composition or lead to the loss of metabolites. In this study, the performance of lyophilization is investigated in...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-05-01
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| Series: | Metabolites |
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| Online Access: | https://www.mdpi.com/2218-1989/13/6/686 |
| _version_ | 1797593533053403136 |
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| author | Christina Maisl Maria Doppler Bernhard Seidl Christoph Bueschl Rainer Schuhmacher |
| author_facet | Christina Maisl Maria Doppler Bernhard Seidl Christoph Bueschl Rainer Schuhmacher |
| author_sort | Christina Maisl |
| collection | DOAJ |
| description | Lyophilization is a common method used for stabilizing biological samples prior to storage or to concentrate extracts. However, it is possible that this process may alter the metabolic composition or lead to the loss of metabolites. In this study, the performance of lyophilization is investigated in the example of wheat roots. To this end, native and <sup>13</sup>C-labelled, fresh or already lyophilized root samples, and (diluted) extracts with dilution factors up to 32 and authentic reference standards were investigated. All samples were analyzed using RP-LC-HRMS. Results show that using lyophilization for the stabilization of plant material altered the metabolic sample composition. Overall, 7% of all wheat metabolites detected in non-lyophilized samples were not detected in dried samples anymore, and up to 43% of the remaining metabolites exhibited significantly increased or decreased abundances. With respect to extract concentration, less than 5% of the expected metabolites were completely lost by lyophilization and the recovery rates of the remaining metabolites were slightly reduced with increasing concentration factors to an average of 85% at an enrichment factor of 32. Compound annotation did not indicate specific classes of wheat metabolites to be affected. |
| first_indexed | 2024-03-11T02:10:40Z |
| format | Article |
| id | doaj.art-a997c1506c2641dd86b8ac9669b59a69 |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-11T02:10:40Z |
| publishDate | 2023-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-a997c1506c2641dd86b8ac9669b59a692023-11-18T11:34:18ZengMDPI AGMetabolites2218-19892023-05-0113668610.3390/metabo13060686Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation TechniqueChristina Maisl0Maria Doppler1Bernhard Seidl2Christoph Bueschl3Rainer Schuhmacher4Department of Agrobiotechnology IFA-Tulln, Institute of Bioanalytics and Agro-Metabolomics, University of Natural Resources and Life Sciences, Vienna, Konrad-Lorenz-Str. 20, 3430 Tulln, AustriaDepartment of Agrobiotechnology IFA-Tulln, Institute of Bioanalytics and Agro-Metabolomics, University of Natural Resources and Life Sciences, Vienna, Konrad-Lorenz-Str. 20, 3430 Tulln, AustriaDepartment of Agrobiotechnology IFA-Tulln, Institute of Bioanalytics and Agro-Metabolomics, University of Natural Resources and Life Sciences, Vienna, Konrad-Lorenz-Str. 20, 3430 Tulln, AustriaDepartment of Agrobiotechnology IFA-Tulln, Institute of Bioanalytics and Agro-Metabolomics, University of Natural Resources and Life Sciences, Vienna, Konrad-Lorenz-Str. 20, 3430 Tulln, AustriaDepartment of Agrobiotechnology IFA-Tulln, Institute of Bioanalytics and Agro-Metabolomics, University of Natural Resources and Life Sciences, Vienna, Konrad-Lorenz-Str. 20, 3430 Tulln, AustriaLyophilization is a common method used for stabilizing biological samples prior to storage or to concentrate extracts. However, it is possible that this process may alter the metabolic composition or lead to the loss of metabolites. In this study, the performance of lyophilization is investigated in the example of wheat roots. To this end, native and <sup>13</sup>C-labelled, fresh or already lyophilized root samples, and (diluted) extracts with dilution factors up to 32 and authentic reference standards were investigated. All samples were analyzed using RP-LC-HRMS. Results show that using lyophilization for the stabilization of plant material altered the metabolic sample composition. Overall, 7% of all wheat metabolites detected in non-lyophilized samples were not detected in dried samples anymore, and up to 43% of the remaining metabolites exhibited significantly increased or decreased abundances. With respect to extract concentration, less than 5% of the expected metabolites were completely lost by lyophilization and the recovery rates of the remaining metabolites were slightly reduced with increasing concentration factors to an average of 85% at an enrichment factor of 32. Compound annotation did not indicate specific classes of wheat metabolites to be affected.https://www.mdpi.com/2218-1989/13/6/686freeze-dryingconcentrationenrichmentstabilizationRP-LC-HRMS |
| spellingShingle | Christina Maisl Maria Doppler Bernhard Seidl Christoph Bueschl Rainer Schuhmacher Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation Technique Metabolites freeze-drying concentration enrichment stabilization RP-LC-HRMS |
| title | Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation Technique |
| title_full | Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation Technique |
| title_fullStr | Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation Technique |
| title_full_unstemmed | Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation Technique |
| title_short | Untargeted Plant Metabolomics: Evaluation of Lyophilization as a Sample Preparation Technique |
| title_sort | untargeted plant metabolomics evaluation of lyophilization as a sample preparation technique |
| topic | freeze-drying concentration enrichment stabilization RP-LC-HRMS |
| url | https://www.mdpi.com/2218-1989/13/6/686 |
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