The DOI matches its Crossref record; 37 of the 37 references it cites were found in a registry or an index.
Metabolomic Analysis via Reversed-Phase Ion-Pairing Liquid Chromatography Coupled to a Stand Alone Orbitrap Mass Spectrometer
Open accessAbstract
We present a liquid chromatography-mass spectrometry (LC-MS) method that capitalizes on the mass-resolving power of the orbitrap to enable sensitive and specific measurement of known and unanticipated metabolites in parallel, with a focus on water-soluble species involved in core metabolism. The reversed phase LC method, with a cycle time 25 min, involves a water-methanol gradient on a C18 column with tributylamine as the ion pairing agent. The MS portion involves full scans from 85 to 1000 m/z at 1 Hz and 100,000 resolution in negative ion mode on a stand alone orbitrap ("Exactive"). The median limit of detection, across 80 metabolite standards, was 5 ng/mL with the linear range typically >or=100-fold. For both standards and a cellular extract from Saccharomyces cerevisiae (Baker's yeast), the median inter-run relative standard deviation in peak intensity was 8%. In yeast exact, we detected 137 known compounds, whose (13)C-labeling patterns could also be tracked to probe metabolic flux. In yeast engineered to lack a gene of unknown function (YKL215C), we observed accumulation of an ion of m/z 128.0351, which we subsequently confirmed to be oxoproline, resulting in annotation of YKL215C as an oxoprolinase. These examples demonstrate the suitability of the present method for quantitative metabolomics, fluxomics, and discovery metabolite profiling.
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Cites
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KEGG for linking genomes to life and the environment
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HMDB: the Human Metabolome Database
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HMDB: a knowledgebase for the human metabolome
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Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli
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Self-citationThe Orbitrap: a new mass spectrometer
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Electrostatic Axially Harmonic Orbital Trapping: A High-Performance Technique of Mass Analysis
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Construction of a set of convenient saccharomyces cerevisiae strains that are isogenic to S288C
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Stincone, A., Prigione, A., Cramer, T., Wamelink, M. M. C., Campbell, K., Cheung, E., Olin‐Sandoval, V., Grüning, N., Krüger, A., Tauqeer Alam, M., et al. (2015) · Biological Reviews · doi:10.1111/brv.12140 · Cited by 1,576
Self-citationQuantitative flux analysis reveals folate-dependent NADPH production
Fan, J., Ye, J., Kamphorst, J. J., Shlomi, T., Thompson, C. B., & Rabinowitz, J. D. (2014) · Nature · doi:10.1038/nature13236 · Cited by 1,114
Self-citationYuan, M., Breitkopf, S. B., Yang, X., & Asara, J. M. (2012) · Nature Protocols · doi:10.1038/nprot.2012.024 · Cited by 1,117
Kamphorst, J. J., Nofal, M., Commisso, C., Hackett, S. R., Lu, W., Grabocka, E., Vander Heiden, M. G., Miller, G., Drebin, J. A., Bar-Sagi, D., et al. (2015) · Cancer Research · doi:10.1158/0008-5472.can-14-2211 · Cited by 893
Self-citationKamphorst, J. J., Cross, J. R., Fan, J., de Stanchina, E., Mathew, R., White, E. P., Thompson, C. B., & Rabinowitz, J. D. (2013) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1307237110 · Cited by 726
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Metadata
- Authors
- Lu, W., Clasquin, M. F., Melamud, E., Amador-Noguez, D., Caudy, A. A., & Rabinowitz, J. D.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2010-03-29
- Volume
- 82
- Issue
- 8
- Pages
- 3212-3221
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W1994124478
- Cited by
- 502
- References
- 37
- FWCI
- 11.11
- Type
- journal article
- Language
- en
- Open access
- green
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- Lu, W., Clasquin, M. F., Melamud, E., Amador-Noguez, D., Caudy, A. A., & Rabinowitz, J. D. (2010). Metabolomic Analysis via Reversed-Phase Ion-Pairing Liquid Chromatography Coupled to a Stand Alone Orbitrap Mass Spectrometer. Analytical Chemistry. https://doi.org/10.1021/ac902837x
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