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Metabolites 2019, 9(2), 32; https://doi.org/10.3390/metabo9020032

Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using 1H-NMR-Based Metabolomics

1
Grice Marine Laboratory, Department of Biology, College of Charleston, 205 Fort Johnson Rd., Charleston, SC 29412, USA
2
Hollings Marine Laboratory, Chemical Science Division, National Institute of Standards and Technology, Charleston, SC 29412, USA
3
Marine Science and Nautical Training Academy (MANTA), 520 Folly Rd., Charleston, SC 29412, USA
4
Hollings Marine Laboratory, National Ocean Service, National Oceanic and Atmospheric Administration, Charleston, SC 29412, USA
*
Author to whom correspondence should be addressed.
Received: 20 December 2018 / Revised: 8 February 2019 / Accepted: 10 February 2019 / Published: 13 February 2019
(This article belongs to the Special Issue Environmental Metabolomics)
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Abstract

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 tert-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 1H NMR metabolomics. View Full-Text
Keywords: metabolomics; method; NMR spectroscopy; sample preparation; coral metabolomics; method; NMR spectroscopy; sample preparation; coral
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Andersson, E.R.; Day, R.D.; Loewenstein, J.M.; Woodley, C.M.; Schock, T.B. Evaluation of Sample Preparation Methods for the Analysis of Reef-Building Corals Using 1H-NMR-Based Metabolomics. Metabolites 2019, 9, 32.

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