Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based Metabolomics
The field of metabolomics generally lacks standardized methods for the preparation of samples prior to analysis. This is especially true for metabolomics of reef-building corals, where the handful of studies that were published employ a range of sample preparation protocols. The utilization of metab...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-02-01
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| Series: | Metabolites |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-1989/9/2/32 |
| _version_ | 1828846581047099392 |
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| author | Erik R. Andersson Rusty D. Day Julie M. Loewenstein Cheryl M. Woodley Tracey B. Schock |
| author_facet | Erik R. Andersson Rusty D. Day Julie M. Loewenstein Cheryl M. Woodley Tracey B. Schock |
| author_sort | Erik R. Andersson |
| collection | DOAJ |
| description | The field of metabolomics generally lacks standardized methods for the preparation of samples prior to analysis. This is especially true for metabolomics of reef-building corals, where the handful of studies that were published employ a range of sample preparation protocols. The utilization of metabolomics may prove essential in understanding coral biology in the face of increasing environmental threats, and an optimized method for preparing coral samples for metabolomics analysis would aid this cause. The current study evaluates three important steps during sample processing of stony corals: (i) metabolite extraction, (ii) metabolism preservation, and (iii) subsampling. Results indicate that a modified Bligh and Dyer extraction is more reproducible across multiple coral species compared to methyl <i>tert</i>-butyl ether and methanol extractions, while a methanol extraction is superior for feature detection. Additionally, few differences were detected between spectra from frozen or lyophilized coral samples. Finally, extraction of entire coral nubbins increased feature detection, but decreased throughput and was more susceptible to subsampling error compared to a novel tissue powder subsampling method. Overall, we recommend the use of a modified Bligh and Dyer extraction, lyophilized samples, and the analysis of brushed tissue powder for the preparation of reef-building coral samples for <sup>1</sup>H NMR metabolomics. |
| first_indexed | 2024-12-12T21:51:46Z |
| format | Article |
| id | doaj.art-a405d87970354c349514c19028376a37 |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-12-12T21:51:46Z |
| publishDate | 2019-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-a405d87970354c349514c19028376a372022-12-22T00:10:46ZengMDPI AGMetabolites2218-19892019-02-01923210.3390/metabo9020032metabo9020032Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based MetabolomicsErik R. Andersson0Rusty D. Day1Julie M. Loewenstein2Cheryl M. Woodley3Tracey B. Schock4Grice Marine Laboratory, Department of Biology, College of Charleston, 205 Fort Johnson Rd., Charleston, SC 29412, USAHollings Marine Laboratory, Chemical Science Division, National Institute of Standards and Technology, Charleston, SC 29412, USAGrice Marine Laboratory, Department of Biology, College of Charleston, 205 Fort Johnson Rd., Charleston, SC 29412, USAHollings Marine Laboratory, National Ocean Service, National Oceanic and Atmospheric Administration, Charleston, SC 29412, USAHollings Marine Laboratory, Chemical Science Division, National Institute of Standards and Technology, Charleston, SC 29412, USAThe field of metabolomics generally lacks standardized methods for the preparation of samples prior to analysis. This is especially true for metabolomics of reef-building corals, where the handful of studies that were published employ a range of sample preparation protocols. The utilization of metabolomics may prove essential in understanding coral biology in the face of increasing environmental threats, and an optimized method for preparing coral samples for metabolomics analysis would aid this cause. The current study evaluates three important steps during sample processing of stony corals: (i) metabolite extraction, (ii) metabolism preservation, and (iii) subsampling. Results indicate that a modified Bligh and Dyer extraction is more reproducible across multiple coral species compared to methyl <i>tert</i>-butyl ether and methanol extractions, while a methanol extraction is superior for feature detection. Additionally, few differences were detected between spectra from frozen or lyophilized coral samples. Finally, extraction of entire coral nubbins increased feature detection, but decreased throughput and was more susceptible to subsampling error compared to a novel tissue powder subsampling method. Overall, we recommend the use of a modified Bligh and Dyer extraction, lyophilized samples, and the analysis of brushed tissue powder for the preparation of reef-building coral samples for <sup>1</sup>H NMR metabolomics.https://www.mdpi.com/2218-1989/9/2/32metabolomicsmethodNMR spectroscopysample preparationcoral |
| spellingShingle | Erik R. Andersson Rusty D. Day Julie M. Loewenstein Cheryl M. Woodley Tracey B. Schock Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based Metabolomics Metabolites metabolomics method NMR spectroscopy sample preparation coral |
| title | Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based Metabolomics |
| title_full | Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based Metabolomics |
| title_fullStr | Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based Metabolomics |
| title_full_unstemmed | Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based Metabolomics |
| title_short | Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using <sup>1</sup>H-NMR-Based Metabolomics |
| title_sort | evaluation of sample preparation methods for the analysis of reef building corals using sup 1 sup h nmr based metabolomics |
| topic | metabolomics method NMR spectroscopy sample preparation coral |
| url | https://www.mdpi.com/2218-1989/9/2/32 |
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