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Metabolites 2013, 3(3), 592-605; doi:10.3390/metabo3030592
Article

Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods

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Received: 17 June 2013; in revised form: 17 July 2013 / Accepted: 17 July 2013 / Published: 23 July 2013
(This article belongs to the Special Issue Sample Preparation for Metabolite Analysis)
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Abstract: Metabolomic and lipidomic approaches aim to measure metabolites or lipids in the cell. Metabolite extraction is a key step in obtaining useful and reliable data for successful metabolite studies. Significant efforts have been made to identify the optimal extraction protocol for various platforms and biological systems, for both polar and non-polar metabolites. Here we report an approach utilizing chemoinformatics for systematic comparison of protocols to extract both from a single sample of the model yeast organism Saccharomyces cerevisiae. Three chloroform/methanol/water partitioning based extraction protocols found in literature were evaluated for their effectiveness at reproducibly extracting both polar and non-polar metabolites. Fatty acid methyl esters and methoxyamine/trimethylsilyl derivatized aqueous compounds were analyzed by gas chromatography mass spectrometry to evaluate non-polar or polar metabolite analysis. The comparative breadth and amount of recovered metabolites was evaluated using multivariate projection methods. This approach identified an optimal protocol consisting of 64 identified polar metabolites from 105 ion hits and 12 fatty acids recovered, and will potentially attenuate the error and variation associated with combining metabolite profiles from different samples for untargeted analysis with both polar and non-polar analytes. It also confirmed the value of using multivariate projection methods to compare established extraction protocols.
Keywords: metabolite extraction; chloroform/methanol/water partitioning; GC-MS; fatty acid methyl esters; polar metabolites; PCA; OPLS; chemoinformatics metabolite extraction; chloroform/methanol/water partitioning; GC-MS; fatty acid methyl esters; polar metabolites; PCA; OPLS; chemoinformatics
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Tambellini, N.P.; Zaremberg, V.; Turner, R.J.; Weljie, A.M. Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods. Metabolites 2013, 3, 592-605.

AMA Style

Tambellini NP, Zaremberg V, Turner RJ, Weljie AM. Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods. Metabolites. 2013; 3(3):592-605.

Chicago/Turabian Style

Tambellini, Nicolas P.; Zaremberg, Vanina; Turner, Raymond J.; Weljie, Aalim M. 2013. "Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods." Metabolites 3, no. 3: 592-605.



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