Stable carbon isotope diagnostics of mammalian metabolism, a high-resolution isotomics approach using amino acid carboxyl groups
Open accessAbstract
The carbon isotopic compositions of amino acids are increasingly measured to characterize diets and metabolic response to diets. We report a new high-resolution system to measure the stable carbon isotopic composition of carboxyl atoms within amino acids. The automated system used HPLC to separate amino acids followed by addition of ninhydrin for decarboxylation and transfer of the evolved CO2 to a stable isotope ratio mass spectrometer for δ13CCARBOXYL measurement. The ninhydrin reaction was conducted at acidic pH (1.5) and elevated temperature (160 oC) giving yields close to 100% for most common amino acids. Eight mammalian keratin samples from herbivores (kudu and caribou), omnivores (humans) and carnivores (bowhead and humpback zooplanktivorous whales) were analysed with this new system. The data provide an initial calibration of reference materials to be used in studies of this type and is the first report of carboxyl carbon isotope distributions in mammals. Results showed widespread 13C enrichments in both essential and non-essential amino acid carboxyl groups, likely linked to decarboxylation of amino acids during normal metabolism. Analyses of non-essential amino acid isotope profiles showed (1) consistent and general taxon-level metabolic differences between the herbivore, human and whale samples, (2) marked differences among individual humans, ruminants and whales (3) evidence for gluconeogenesis in the wildlife samples, and (4) extensive 13C enrichment likely associated with fasting in the humpback whale sample. Future mammalian research related to the metabolism of growth, reproduction, aging and disease may benefit from using this technique. Values obtained for internationally available samples USGS42 and USGS43 (Tibetan and Indian human hair) provide a first characterization of reference materials for δ13CCARBOXYL profiles.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Pyruvate entering the citric cycle paves the way for glutamate, glutamine, asparagine, aspartate, and proline biosynthesis.
“Furthermore, being introduced into the citric cycle, pyruvate paves the way for glutamate, glutamine, asparagine, aspartate, and proline biosynthesis [48].”
From 20-Hydroxyecdysone Boosts Energy Production and Biosynthetic Processes in Non-Transformed Mouse Cells (2024) · cites this paper as [48]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
[117] Chromatographic determination of amino acids by the use of automatic recording equipment
Moore, S., & Stein, W. H. (1963) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(63)06257-1 · Cited by 1,701 · see citing works
Mechanism of Carbon Isotope Fractionation Associated with Lipid Synthesis
DeNiro, M. J., & Epstein, S. (1977) · Science · doi:10.1126/science.327543 · Cited by 1,461 · see citing works
Fractionation of Carbon and Hydrogen Isotopes in Biosynthetic Processes
Hayes, J. M. (2001) · Reviews in Mineralogy and Geochemistry · doi:10.2138/gsrmg.43.1.225 · Cited by 792 · see citing works
Wassenaar, L. I., & Hobson, K. A. (2003) · Isotopes in Environmental and Health Studies · doi:10.1080/1025601031000096781 · Cited by 669 · see citing works
Hamilton, P., & Slyke, D. D. V. (1943) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)51268-0 · Cited by 598 · see citing works
CARBON ISOTOPE FRACTIONATION IN FORMATION OF AMINO ACIDS BY PHOTOSYNTHETIC ORGANISMS
Abelson, P. H., & Hoering, T. C. (1961) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.47.5.623 · Cited by 501 · see citing works
GASOMETRIC DETERMINATION OF CARBOXYL GROUPS IN FREE AMINO ACIDS
Slyke, D. D. V., Dillon, R. T., MacFadyen, D. A., et al. (1941) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)72810-x · Cited by 459 · see citing works
Metabolic fractionation of C13 & C12 in plants
Park, R. B., & Epstein, S. (1961) · PLANT PHYSIOLOGY · doi:10.1104/pp.36.2.133 · Cited by 399 · see citing works
A new concept for isotope ratio monitoring liquid chromatography/mass spectrometry
Krummen, M., Hilkert, A. W., Juchelka, D., et al. (2004) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1620 · Cited by 268 · see citing works
Melzer, E. W., & Schmidt, H. (1987) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)47543-6 · Cited by 208 · see citing works
Newsome, S. D., Fogel, M. L., Kelly, L., & del Rio, C. M. (2011) · Functional Ecology · doi:10.1111/j.1365-2435.2011.01866.x · Cited by 130 · see citing works
Silfer, J. A., Engel, M. H., & Macko, S. A. (1992) · Chemical Geology Isotope Geoscience section · doi:10.1016/0009-2541(92)90003-n · Cited by 121 · see citing works
Newsome, S. D., Wolf, N., Peters, J. M., & Fogel, M. L. (2014) · Integrative and Comparative Biology · doi:10.1093/icb/icu106 · Cited by 117 · see citing works
Isotopic fractionation during peptide bond hydrolysis
Bada, J. L., Schoeninger, M. J., & Schimmelmann, A. (1989) · Geochimica et Cosmochimica Acta · doi:10.1016/0016-7037(89)90114-2 · Cited by 105 · see citing works
Eisenmann, P., Fry, B., Holyoake, C. S., Coughran, D. K., Nicol, S., & Nash, S. B. (2016) · PLoS ONE · doi:10.1371/journal.pone.0156698 · Cited by 105 · see citing works
Self-citationCarbon isotope systematics of a mantle “hotspot”: a comparison of Loihi Seamount and MORB glasses
Exley, R. A., Mattey, D. P., Clague, D. A., & Pillinger, C. T. (1986) · Earth and Planetary Science Letters · doi:10.1016/0012-821x(86)90060-9 · Cited by 104 · see citing works
Smith, C. I., Fuller, B. T., Choy, K., et al. (2009) · Analytical Biochemistry · doi:10.1016/j.ab.2009.04.014 · Cited by 98 · see citing works
Coplen, T. B., & Qi, H. (2011) · Forensic Science International · doi:10.1016/j.forsciint.2011.07.035 · Cited by 97 · see citing works
Block, R. J. (1939) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)73740-x · Cited by 87 · see citing works
Matthews, C., & Ferguson, S. (2014) · Marine Ecology Progress Series · doi:10.3354/meps11145 · Cited by 68 · see citing works
δ15N Value Does Not Reflect Fasting in Mysticetes
Aguilar, Á., Giménez, J., Gómez‐Campos, E., et al. (2014) · PLoS ONE · doi:10.1371/journal.pone.0092288 · Cited by 67 · see citing works
McCullagh, J. S. O., Gaye‐Siessegger, J., & Focken, U. (2008) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.3554 · Cited by 67 · see citing works
Choy, K., Nash, S. H., Kristal, A. R., Hopkins, S. E., Boyer, B. B., & O’Brien, D. M. (2013) · Journal of Nutrition · doi:10.3945/jn.112.172999 · Cited by 66 · see citing works
Choy, K., Smith, C. I., Fuller, B. T., et al. (2010) · Geochimica et Cosmochimica Acta · doi:10.1016/j.gca.2010.07.025 · Cited by 64 · see citing works
Brenna, J. T. (2001) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.325 · Cited by 62 · see citing works
Isotopic and Elemental Composition of Roasted Coffee as a Guide to Authenticity and Origin
Carter, J. F., Yates, H. S. A., & Tinggi, U. (2015) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.5b01526 · Cited by 48 · see citing works
Self-citationMetges, C. C., & Daenzer, M. (2000) · Analytical Biochemistry · doi:10.1006/abio.1999.4426 · Cited by 48 · see citing works
THE AMINO ACID CONTENT OF SOME SCLEROPROTEINS
GRAHAM, C. E., Waitkoff, H. K., & Hier, S. W. (1949) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)56995-7 · Cited by 47 · see citing works
In Silico Evidence for Gluconeogenesis from Fatty Acids in Humans
Kaleta, C., de Figueiredo, L. F., Werner, S., Guthke, R., Ristow, M., & Schuster, S. (2011) · PLoS Computational Biology · doi:10.1371/journal.pcbi.1002116 · Cited by 43 · see citing works
Ab initio calculation of the carbon isotope signatures of amino acids
Rustad, J. R. (2009) · Organic Geochemistry · doi:10.1016/j.orggeochem.2009.03.003 · Cited by 36 · see citing works
Larsen, T., Ventura, M., O’Brien, D. M., Magid, J., Lomstein, B. A., & Larsen, J. (2010) · Soil Biology and Biochemistry · doi:10.1016/j.soilbio.2010.12.008 · Cited by 36 · see citing works
Jim, S., Jones, V., Copley, M. S., Ambrose, S. H., & Evershed, R. P. (2003) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1177 · Cited by 33 · see citing works
Amino acid composition of human scalp hair as a biometric classifier and investigative lead
Rashaid, A. H. B., Harrington, P. D. B., & Jackson, G. P. (2014) · Analytical Methods · doi:10.1039/c4ay02588a · Cited by 28 · see citing works
Fry, B., Carter, J. F., Yamada, K., et al. (2018) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.8126 · Cited by 28 · see citing works
Self-citationAdeniyi-Jones, S. K., & Karnovsky, M. L. (1981) · Journal of Clinical Investigation · doi:10.1172/jci110264 · Cited by 21 · see citing works
Methods for improving the visualization and deconvolution of isotopic signals
Tcherkez, G. G. B., Ghashghaie, J., & Griffiths, H. (2007) · Plant Cell & Environment · doi:10.1111/j.1365-3040.2007.01687.x · Cited by 16 · see citing works
Godin, J., Ross, A. B., Rezzi, S., Poussin, C., Martin, F., Fuerholz, A., Cléroux, M., Mermoud, A., Tornier, L., Vera, F. A., et al. (2009) · Analytical Chemistry · doi:10.1021/ac902086g · Cited by 14 · see citing works
Hair Amino Acids: Normal Values and Results in Metabolic Errors
SANDE, M. V. (1970) · Archives of Disease in Childhood · doi:10.1136/adc.45.243.678 · Cited by 14 · see citing works
Savidge, W. B., & Blair, N. E. (2004) · Journal of Experimental Marine Biology and Ecology · doi:10.1016/j.jembe.2004.02.010 · Cited by 7 · see citing works
[Isotopic 13 C- 12 C relation in a malignant tumor in the rat].
Mosora, F., Lacroix, M., & Duchesne, J. (1972) · PubMed · Cited by 2 · see citing works
[Free radicals and C13-C12 ratio in plant tumors].
Mosora, F., Maréchal, R., Manil, P., et al. (1972) · PubMed · Cited by 1 · see citing works
Eisenmann, P. (2016) · Griffith Research Online · doi:10.25904/1912/2987 · Cited by 1 · see citing works
Flagged first, then by citations
Research works printed but not linked: 3
- [1]Mosora F, Marechal R, Manil P, Duchesne J. Sur le rapport isotopique 13C/12C dans une tumeur ajligne chez le Rat. C R Acad Sci Hebd Seances Acad Sci D. 1972; 275(4): 599– 601. 4342849
- [2]Mosora F, Lacroix M, Duchesne J. Radicaux libres et rapport isotopique 13C/12C dans des tumeurs vegetales. C R Acad Sci Hebd Seances Acad Sci D. 1972; 275(15): 1701– 2. 4629943
- [7]Meier-Augenstein W. Stable isotope forensics: methods and forensic applications of stable isotope analysis 2nd ed. Hoboken, NJ: John Wiley & Sons,; 2018.
Cited by
Sorted by citations
Shuvalov, O., Kirdeeva, Y., Fefilova, E., Daks, A., Fedorova, O., Parfenyev, S., Nazarov, A., Vlasova, Y., Krasnov, G. S., & Barlev, N. A. (2024) · Antioxidants · doi:10.3390/antiox13111349 · Cited by 9 · see citing works
- Authors
- Fry, B., & Carter, J. F.
- Affiliations
- Griffith University (AU)
- Queensland Health (AU)
- Corresponding author
- Fry, Brian
- Venue
- PLOS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2019-10-28
- Volume
- 14
- Issue
- 10
- Pages
- e0224297
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC6816566
- OpenAlex
- W2982472422
- Cited by
- 22
- References
- 42
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 2 · 2023: 2 · 2022: 3 · 2021: 4 · 2020: 5
- FWCI
- 1.90
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Figshare (submittedVersion, cc-by)
- Griffith Research Online (submittedVersion, cc-by)
- Funders
- Australian Research Council (DP150104006)
- Department of Health, Queensland (RSS15-005)
- Topics
- Physical Sciences › Environmental Science › Ecology › Isotope Analysis in Ecology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Physical Sciences › Earth and Planetary Sciences › Paleontology › Archaeology and ancient environmental studies
- Keywords
- stable carbon isotopes, mammalian metabolism, keratin, herbivores, carnivores, humpback whale, Kudus, reindeer, bowhead whale, gluconeogenesis, fasting, reference materials, diet and metabolism
- MeSH terms
- Amino Acids, Animals, Carbon Isotopes, Chromatography, High Pressure Liquid, Hydrolysis, Mass Spectrometry, Metabolomics
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
