A Stable-Isotope Mass Spectrometry-Based Metabolic Footprinting Approach to Analyze Exudates from Phytoplankton
Abstract
:1. Introduction
2. Results and Discussion
2.1. High-Resolution Mass Spectrometry-Based Metabolic Footprinting
2.2. Locating Stable Isotope Patterns
m/z (all-12C containing peak as [M − H]− ion) | Empirical formula | Labelling Efficiency (%) | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
r-value | p-value | ||||||||||
50% | 55% | 60% | 65% | 70% | 50% | 55% | 60% | 65% | 70% | ||
125.06084 | 12C7H10O2 | 0.86 | 0.99 | 0.93 | 0.68 | 0.35 | 0.0289 | 0.0001 | 0.0080 | 0.1347 | 0.4913 |
131.07140 | 12C6H12O3 | 0.92 | 0.99 | 0.82 | 0.48 | 0.12 | 0.0244 | 0.0010 | 0.0918 | 0.4181 | 0.8532 |
227.20143 | 12C14H28O2 | 0.63 | 0.90 | 1.00 | 0.88 | 0.58 | 0.0369 | 0.0002 | 0.0000 | 0.0004 | 0.0615 |
2.3. Estimation of the Stable Isotope Labelling Efficiency
2.4. Putative Annotation of Exuded Metabolites
m/z | Colour (in Figure 5) | Empirical Formula (peak) | Ion Form | Theoretical Mass (Da) | Mass Error (ppm) | KEGG Compound | r-value | p-value |
---|---|---|---|---|---|---|---|---|
185.11855 | blue | C10H18O3 | [M − H]− | 185.11832 | 1.25 | [(3R)-6-Hydroxy-3-isopropenyl-heptanoate, (3S)-6-Hydroxy-3-isopropenyl-heptanoate, (5R)-6-Hydroxy-5-isopropenyl-2-methylhexanoate, (5S)-6-Hydroxy-5-isopropenyl-2-methylhexanoate, 10-Oxodecanoate, 2-Oxodecanoic acid, Epomediol] | 0.95 | 0.0115 |
green | C8H14O | [M + Acetate]− | 185.11832 | 1.25 | [Sulcatone] | −0.09 | 0.9149 | |
197.15491 | blue | C12H22O2 | [M − H]− | 197.15470 | 1.05 | [(−)-Menthyl acetate, Citronellyl acetate, Decanoyl acetaldehyde, Neomenthyl acetate] | 1.00 | 0.0000 |
green | C10H18 | [M + Acetate]− | 197.15470 | 1.05 | 0.58 | 0.1735 | ||
409.29594 | red | C22H38O3 | [M + Acetate]− | 409.29595 | −0.02 | 0.99 | 0.0000 | |
blue | C21H45N2O2P | [M + Acetate]− | 409.29653 | −1.45 | 0.91 | 0.0000 | ||
orange | C24H42O5 | [M − H]− | 409.29595 | −0.02 | 0.91 | 0.0000 | ||
green | C19H39N8P | [M − H]− | 409.29625 | −0.77 | 0.55 | 0.0506 |
3. Experimental Section
3.1. Cell Culture and Stable Isotope Labelling
3.2. Metabolite Extraction from Media
3.3. FT-ICR MS Analysis, Spectral Processing and Unsupervised Multivariate Analyses
3.4. Locating and Putative Identification of SIPs
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Weber, R.J.M.; Selander, E.; Sommer, U.; Viant, M.R. A Stable-Isotope Mass Spectrometry-Based Metabolic Footprinting Approach to Analyze Exudates from Phytoplankton. Mar. Drugs 2013, 11, 4158-4175. https://doi.org/10.3390/md11114158
Weber RJM, Selander E, Sommer U, Viant MR. A Stable-Isotope Mass Spectrometry-Based Metabolic Footprinting Approach to Analyze Exudates from Phytoplankton. Marine Drugs. 2013; 11(11):4158-4175. https://doi.org/10.3390/md11114158
Chicago/Turabian StyleWeber, Ralf J. M., Erik Selander, Ulf Sommer, and Mark R. Viant. 2013. "A Stable-Isotope Mass Spectrometry-Based Metabolic Footprinting Approach to Analyze Exudates from Phytoplankton" Marine Drugs 11, no. 11: 4158-4175. https://doi.org/10.3390/md11114158