Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods
Abstract
:1. Introduction
2. Results and Discussion
2.1. Unsupervised Analysis Clearly Differentiates Extraction Method
2.2. Supervised Analysis Identifies 36 Metabolites and Four Fatty Acids Differentiating the Extraction Methods
Metabolite | VIP | Coefficient Extraction 1 | Coefficient Extraction 2 | Coefficient Extraction 3 |
---|---|---|---|---|
Threonine | 1.832 | 0.013 | 0.046 | −0.059 |
Glycerol | 1.703 | −0.012 | −0.042 | 0.054 |
Phenylalanine | 1.678 | 0.030 | 0.034 | −0.064 |
Alanine-3cyano | 1.666 | 0.037 | 0.009 | −0.046 |
Methionine | 1.649 | 0.014 | 0.041 | −0.055 |
Proline | 1.630 | −0.001 | 0.029 | −0.028 |
Sorbitol | 1.603 | 0.002 | 0.023 | −0.024 |
Phosphoric Acid | 1.575 | −0.008 | 0.052 | −0.043 |
Homoserine | 1.531 | −0.001 | 0.049 | −0.048 |
Pyroglutamic Acid | 1.496 | 0.002 | 0.014 | −0.016 |
Alanine | 1.476 | −0.023 | 0.025 | −0.002 |
Ornithine | 1.461 | −0.011 | −0.046 | 0.057 |
Serine-O acetyl | 1.382 | −0.029 | −0.035 | 0.064 |
Fumaric Acid | 1.372 | 0.002 | 0.004 | −0.006 |
Trehalose-alpha,alpha’-D | 1.340 | 0.013 | 0.002 | −0.015 |
Alanine-beta | 1.327 | 0.007 | 0.006 | −0.013 |
Succinic Acid | 1.310 | −0.006 | −0.011 | 0.017 |
Malic acid, 2-isopropyl | 1.299 | 0.016 | 0.020 | −0.035 |
Decan-1-ol, n- | 1.296 | 0.003 | −0.004 | 0.002 |
Glycine | 1.271 | −0.027 | −0.014 | 0.041 |
Valine | 1.267 | −0.010 | 0.033 | −0.023 |
Aspartic Acid | 1.261 | 0.016 | 0.012 | −0.028 |
Arginine [-NH3] | 1.259 | 0.030 | 0.003 | −0.033 |
Glutamic Acid | 1.213 | 0.009 | 0.007 | −0.016 |
Hexadecane, n- | 1.210 | −0.016 | 0.036 | −0.019 |
Malic Acid | 1.190 | 0.005 | 0.002 | −0.006 |
Uracil | 1.189 | −0.039 | −0.017 | 0.056 |
Isoleucine | 1.162 | −0.009 | 0.019 | −0.011 |
Glutamine, DL- | 1.156 | 0.029 | −0.023 | −0.007 |
Octylamine | 1.131 | −0.005 | 0.005 | −0.001 |
Tyramine | 1.119 | 0.014 | 0.012 | −0.026 |
Butanoic Acid | 1.063 | −0.010 | 0.019 | −0.008 |
Serine | 1.062 | −0.016 | 0.036 | −0.020 |
Pentadecane, n- | 1.029 | −0.015 | 0.022 | −0.008 |
Citric Acid | 1.024 | 0.018 | 0.005 | −0.023 |
Heptadecan-1-ol | 1.010 | −0.011 | 0.011 | −0.001 |
Palmitic Acid | 1.950 | 0.581 | −0.372 | −0.182 |
Palmitoleic Acid | 1.273 | 0.173 | 0.082 | −0.221 |
Oleic Acid | 1.239 | 0.123 | 0.066 | −0.164 |
Stearic Acid | 1.139 | 0.058 | −0.197 | 0.121 |
2.3. Comparison of FAME and Aqueous Metabolite Profiles Obtained
2.4. Summary
3. Experimental Section
3.1. Yeast Growth and Harvesting
3.2. Metabolite Extraction
3.2.1. Extraction Protocol 1
3.2.2. Extraction Protocol 2
3.2.3. Extraction Protocol 3
3.3. Derivatization and Sample Preparation
3.4. GC-MS Data Acquisition
3.5. Data Processing and Interpretation
4. Conclusions
Acknowledgments
Conflict of Interest
References
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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. https://doi.org/10.3390/metabo3030592
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. https://doi.org/10.3390/metabo3030592
Chicago/Turabian StyleTambellini, Nicolas P., Vanina Zaremberg, Raymond J. Turner, and Aalim M. Weljie. 2013. "Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods" Metabolites 3, no. 3: 592-605. https://doi.org/10.3390/metabo3030592