Identification of 4-deoxythreonic Acid Present in Human Urine Using HPLC and NMR Techniques

Overview

Journal J Pharm Biomed Anal

Specialty Chemistry

Date 2009 Jul 21

PMID 19615840

Citations 16

Authors

Emmanuel Appiah-Amponsah

Narasimhamurthy Shanaiah

G A Nagana Gowda

Kwadwo Owusu-Sarfo

Tao Ye

Daniel Raftery

Affiliations

Soon will be listed here.

Abstract

The 1H NMR spectrum of urine exhibits a large number of detectable and quantifiable metabolites and hence urine metabolite profiling is potentially useful for the study of systems biology and the discovery of biomarkers for drug development or clinical applications. While a number of metabolites (50-100) are readily detectable in urine by NMR, a much larger number is potentially available if lower concentration species can be detected unambiguously. Lower concentration metabolites are thought to be more specific to certain disease states and thus it is important to detect these metabolites with certainty. We report the identification of 4-deoxythreonic acid, a relatively low concentration endogenous metabolite that has not been previously identified in the 1H NMR spectrum of human urine. The use of HPLC and NMR spectroscopy facilitated the unequivocal and non-invasive identification of the molecule in urine which is complicated by extensive peak overlap and multiple, similar resonances from other metabolites such as 3-hydroxybutanoic acid. High-resolution detection and good sensitivity were achieved by the combination of multiple chromatographic fraction collection, sample pre-concentration, and the use of a cryogenically cooled NMR probe.

Citing Articles

The multi-omics signatures of telomere length in childhood.

Wang C, Martens D, Bustamante M, Alfano R, Plusquin M, Maitre L BMC Genomics. 2025; 26(1):75.

PMID: 39871190 PMC: 11771044. DOI: 10.1186/s12864-025-11209-5.

Urinary metabolomics provide insights into coronary artery disease in individuals with type 1 diabetes.

Antikainen A, Mutter S, Harjutsalo V, Thorn L, Groop P, Sandholm N Cardiovasc Diabetol. 2024; 23(1):425.

PMID: 39593124 PMC: 11590341. DOI: 10.1186/s12933-024-02512-8.

Urine Metabolomics during a Legume Diet Intervention Suggests Altered Metabolic Signatures and Potential New Intake Markers: First Insights.

Ferreira H, Duarte D, Rodrigues J, Vasconcelos M, Pinto E, Gil A ACS Omega. 2024; 9(43):43453-43468.

PMID: 39494014 PMC: 11525520. DOI: 10.1021/acsomega.4c04795.

Human cytosolic transaminases: side activities and patterns of discrimination towards physiologically available alternative substrates.

Caligiore F, Zangelmi E, Vetro C, Kentache T, Dewulf J, Veiga-da-Cunha M Cell Mol Life Sci. 2022; 79(8):421.

PMID: 35834009 PMC: 9283133. DOI: 10.1007/s00018-022-04439-3.

Metabolic adjustments of blood-stage Plasmodium falciparum in response to sublethal pyrazoleamide exposure.

Tewari S, Kwan B, Elahi R, Rajaram K, Reifman J, Prigge S Sci Rep. 2022; 12(1):1167.

PMID: 35064153 PMC: 8782945. DOI: 10.1038/s41598-022-04985-7.

References

Kassel D, Martin M, Schall W, Sweeley C . Urinary metabolites of L-threonine in type 1 diabetes determined by combined gas chromatography/chemical ionization mass spectrometry. Biomed Environ Mass Spectrom. 1986; 13(10):535-40. DOI: 10.1002/bms.1200131004. View

Lenz E, Williams R, Sidaway J, Smith B, Plumb R, Johnson K . The application of microbore UPLC/oa-TOF-MS and 1H NMR spectroscopy to the metabonomic analysis of rat urine following the intravenous administration of pravastatin. J Pharm Biomed Anal. 2007; 44(4):845-52. DOI: 10.1016/j.jpba.2007.04.035. View

Gowda G, Zhang S, Gu H, Asiago V, Shanaiah N, Raftery D . Metabolomics-based methods for early disease diagnostics. Expert Rev Mol Diagn. 2008; 8(5):617-33. PMC: 3890417. DOI: 10.1586/14737159.8.5.617. View

Djukovic D, Liu S, Henry I, Tobias B, Raftery D . Signal enhancement in HPLC/microcoil NMR using automated column trapping. Anal Chem. 2006; 78(20):7154-60. PMC: 2577147. DOI: 10.1021/ac0605748. View

Godejohann M . Hydrophilic interaction chromatography coupled to nuclear magnetic resonance spectroscopy and mass spectroscopy--a new approach for the separation and identification of extremely polar analytes in bodyfluids. J Chromatogr A. 2006; 1156(1-2):87-93. DOI: 10.1016/j.chroma.2006.10.053. View

Graca G, Duarte I, Goodfellow B, Carreira I, Couceiro A, Domingues M . Metabolite profiling of human amniotic fluid by hyphenated nuclear magnetic resonance spectroscopy. Anal Chem. 2008; 80(15):6085-92. DOI: 10.1021/ac800907f. View

Nicholls A, Farrant R, Shockcor J, Unger S, Wilson I, Lindon J . NMR and HPLC-NMR spectroscopic studies of futile deacetylation in paracetamol metabolites in rat and man. J Pharm Biomed Anal. 1997; 15(7):901-10. DOI: 10.1016/s0731-7085(96)01950-4. View

Pan Z, Gu H, Talaty N, Chen H, Shanaiah N, Hainline B . Principal component analysis of urine metabolites detected by NMR and DESI-MS in patients with inborn errors of metabolism. Anal Bioanal Chem. 2006; 387(2):539-49. DOI: 10.1007/s00216-006-0546-7. View

Djukovic D, Appiah-Amponsah E, Shanaiah N, Gowda G, Henry I, Everly M . Ibuprofen metabolite profiling using a combination of SPE/column-trapping and HPLC-micro-coil NMR. J Pharm Biomed Anal. 2008; 47(2):328-34. DOI: 10.1016/j.jpba.2007.12.035. View

10.

Kind T, Tolstikov V, Fiehn O, Weiss R . A comprehensive urinary metabolomic approach for identifying kidney cancerr. Anal Biochem. 2007; 363(2):185-95. DOI: 10.1016/j.ab.2007.01.028. View

11.

Ballevre O, Prugnaud J, Houlier M, Arnal M . Assessment of threonine metabolism in vivo by gas chromatography/mass spectrometry and stable isotope infusion. Anal Biochem. 1991; 193(2):212-9. DOI: 10.1016/0003-2697(91)90011-h. View

12.

Shockcor J, Unger S, Wilson I, Foxall P, Nicholson J, Lindon J . Combined HPLC, NMR spectroscopy, and ion-trap mass spectrometry with application to the detection and characterization of xenobiotic and endogenous metabolites in human urine. Anal Chem. 1996; 68(24):4431-5. DOI: 10.1021/ac9606463. View

13.

Thompson J, Markey S, Fennessey P . Gas-chromatographic/mass-spectrometric identification and quantitation of tetronic and deoxytetronic acids in urine from normal adults and neonates. Clin Chem. 1975; 21(13):1892-8. View

14.

Zhang S, Gowda G, Asiago V, Shanaiah N, Barbas C, Raftery D . Correlative and quantitative 1H NMR-based metabolomics reveals specific metabolic pathway disturbances in diabetic rats. Anal Biochem. 2008; 383(1):76-84. PMC: 3867451. DOI: 10.1016/j.ab.2008.07.041. View

15.

Bales J, Sadler P, Nicholson J, Timbrell J . Urinary excretion of acetaminophen and its metabolites as studied by proton NMR spectroscopy. Clin Chem. 1984; 30(10):1631-6. View

16.

Sandusky P, Raftery D . Use of semiselective TOCSY and the pearson correlation for the metabonomic analysis of biofluid mixtures: application to urine. Anal Chem. 2005; 77(23):7717-23. DOI: 10.1021/ac0510890. View

17.

Lauridsen M, Hansen S, Jaroszewski J, Cornett C . Human urine as test material in 1H NMR-based metabonomics: recommendations for sample preparation and storage. Anal Chem. 2007; 79(3):1181-6. DOI: 10.1021/ac061354x. View

18.

Sandusky P, Raftery D . Use of selective TOCSY NMR experiments for quantifying minor components in complex mixtures: application to the metabonomics of amino acids in honey. Anal Chem. 2005; 77(8):2455-63. DOI: 10.1021/ac0484979. View