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Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research
Many metabolomics studies use mixtures of (acidified) methanol and water for sample extraction. In the present study, we investigated if the extraction with methanol can result in artifacts. To this end, wheat leaves were extracted with mixtures of native and deuterium-labeled methanol and water, wi...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5875991/ https://www.ncbi.nlm.nih.gov/pubmed/29271872 http://dx.doi.org/10.3390/metabo8010001 |
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author | Sauerschnig, Claudia Doppler, Maria Bueschl, Christoph Schuhmacher, Rainer |
author_facet | Sauerschnig, Claudia Doppler, Maria Bueschl, Christoph Schuhmacher, Rainer |
author_sort | Sauerschnig, Claudia |
collection | PubMed |
description | Many metabolomics studies use mixtures of (acidified) methanol and water for sample extraction. In the present study, we investigated if the extraction with methanol can result in artifacts. To this end, wheat leaves were extracted with mixtures of native and deuterium-labeled methanol and water, with or without 0.1% formic acid. Subsequently, the extracts were analyzed immediately or after storage at 10 °C, −20 °C or −80 °C with an HPLC-HESI-QExactive HF-Orbitrap instrument. Our results showed that 88 (8%) of the >1100 detected compounds were derived from the reaction with methanol and either formed during sample extraction or short-term storage. Artifacts were found for various substance classes such as flavonoids, carotenoids, tetrapyrrols, fatty acids and other carboxylic acids that are typically investigated in metabolomics studies. 58 of 88 artifacts were common between the two tested extraction variants. Remarkably, 34 of 73 (acidified extraction solvent) and 33 of 73 (non-acidified extraction solvent) artifacts were formed de novo as none of these meth(ox)ylated metabolites were found after extraction of native leaf samples with CD(3)OH/H(2)O. Moreover, sample extracts stored at 10 °C for several days, as can typically be the case during longer measurement sequences, led to an increase in both the number and abundance of methylated artifacts. In contrast, frozen sample extracts were relatively stable during a storage period of one week. Our study shows that caution has to be exercised if methanol is used as the extraction solvent as the detected metabolites might be artifacts rather than natural constituents of the biological system. In addition, we recommend storing sample extracts in deep freezers immediately after extraction until measurement. |
format | Online Article Text |
id | pubmed-5875991 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-58759912018-03-30 Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research Sauerschnig, Claudia Doppler, Maria Bueschl, Christoph Schuhmacher, Rainer Metabolites Article Many metabolomics studies use mixtures of (acidified) methanol and water for sample extraction. In the present study, we investigated if the extraction with methanol can result in artifacts. To this end, wheat leaves were extracted with mixtures of native and deuterium-labeled methanol and water, with or without 0.1% formic acid. Subsequently, the extracts were analyzed immediately or after storage at 10 °C, −20 °C or −80 °C with an HPLC-HESI-QExactive HF-Orbitrap instrument. Our results showed that 88 (8%) of the >1100 detected compounds were derived from the reaction with methanol and either formed during sample extraction or short-term storage. Artifacts were found for various substance classes such as flavonoids, carotenoids, tetrapyrrols, fatty acids and other carboxylic acids that are typically investigated in metabolomics studies. 58 of 88 artifacts were common between the two tested extraction variants. Remarkably, 34 of 73 (acidified extraction solvent) and 33 of 73 (non-acidified extraction solvent) artifacts were formed de novo as none of these meth(ox)ylated metabolites were found after extraction of native leaf samples with CD(3)OH/H(2)O. Moreover, sample extracts stored at 10 °C for several days, as can typically be the case during longer measurement sequences, led to an increase in both the number and abundance of methylated artifacts. In contrast, frozen sample extracts were relatively stable during a storage period of one week. Our study shows that caution has to be exercised if methanol is used as the extraction solvent as the detected metabolites might be artifacts rather than natural constituents of the biological system. In addition, we recommend storing sample extracts in deep freezers immediately after extraction until measurement. MDPI 2017-12-22 /pmc/articles/PMC5875991/ /pubmed/29271872 http://dx.doi.org/10.3390/metabo8010001 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Sauerschnig, Claudia Doppler, Maria Bueschl, Christoph Schuhmacher, Rainer Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research |
title | Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research |
title_full | Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research |
title_fullStr | Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research |
title_full_unstemmed | Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research |
title_short | Methanol Generates Numerous Artifacts during Sample Extraction and Storage of Extracts in Metabolomics Research |
title_sort | methanol generates numerous artifacts during sample extraction and storage of extracts in metabolomics research |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5875991/ https://www.ncbi.nlm.nih.gov/pubmed/29271872 http://dx.doi.org/10.3390/metabo8010001 |
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