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Analysis of human urine metabolites using SPE and NMR spectroscopy

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Abstract

Nuclear magnetic resonance (NMR) spectroscopic analysis of metabonome/metabolome has wide-spread applications in biomedical science researches. However, most of NMR resonances for urinary metabolites remain to be fully assigned. In the present study, human urine samples from two healthy volunteers were pre-treated with C18 solid-phase extraction and the resultant 5 sub-fractions were subjected to one-and two-dimensional NMR studies, including 1H J-Resolved, 1H-1H COSY, 1H-1H TOCSY, 1H-13C HSQC, and HMBC 2D NMR. More than 70 low molecular weight metabolites were identified, and complete assignments of 1H and 13C resonances including many complex coupled spin systems were obtained.

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References

  1. 1

    Nicholson J K, Lindon J C, Holmes E. ’Metabonomics’: Understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica, 1999, 29(11): 1181–1189

  2. 2

    Tang H R, Wang Y L. Metabonomics: A revolution in progress. Prog Biochem Biophys, 2006, 33(5): 401–417

  3. 3

    Tang H R, Wang Y L. Nutrimetabonomics-metabonomics in food science. In: Magnetic Resonance in Food Science: From Molecules to Man (eds. Farhat I A, Belton P S, and Webb G A). Cambridge: Royal Society of Chemistry, 2007

  4. 4

    Tang H R, Wang Y L. High Resolution NMR Spectroscopy in human metabolism and metabonomics. In: Modern Magnetic Resonance, London: Springer-Verlag, 2006

  5. 5

    Holmes E, Tang H R, Wang Y L, Seger C. The assessment of plant metabolite profiles by NMR-based methodologies. Plant Med, 2006, 72(9): 771–785

  6. 6

    Smith L M, Maher A D, Cloarec O, Rantalainen M, Tang H R, Elliott P, Stamler J, Lindon J C, Holmes E, Nicholson J K. Statistical correlation and projection methods for improved information recovery from diffusion-edited NMR spectra of biological samples. Anal Chem, 2007, 79(15): 5682–5689

  7. 7

    Crockford D J, Holmes E, Lindon J C, Plumb R S, Zirah S, Bruce S J, Rainville P, Stumpf C L, Nocholson J K. Statistical heterospectroscopy, an approach to the integrated analysis of NMR and UPLC-MS data sets: Application in metabonomic toxicology studies. Anal Chem, 2006, 78(2): 363–371

  8. 8

    Rantalainen M, Cloarec O, Beckonert O, Wilson D J, Tonge R, Rowlinson R, Rayner S, Nickson J, Wilkinson R W, Mills J D, Trygg J, Nicholson J K, Holmes E. Statistically integrated metabonomic-proteomic studies on a human prostate cancer xenograft model in mice. J Proteom Res, 2006, 5(10): 2642–2655

  9. 9

    Yap I K S, Clayton T A, Tang H R, Everett J R, Hanton G, Provost J P, Net J L, Charuel C, Lindon J C, Nicholson J K. An integrated metabonomic approach to describe temporal metabolic disregulation induced in the rat by the model hepatotoxin allyl formate. J Proteom Res, 2006, 5(10): 2675–2684

  10. 10

    Plumb R S, Johnson K A, Rainville P, Smith I D, Castro-Perez J M, Nicholson J K. UPLC/MSE. A new approach for generating molecular fragment information for biomarker structure elucidation. Rapid Commun Mass Spectr, 2006, 20(13): 1989–1994

  11. 11

    Crockford D J, Lindon J C, Cloarec O, Plumb R S, Bruce S J, Zirah S, Rainville P, Stumpf C L, Johnson S K, Holmes J E, Nicholson J K. Statistical search space reduction and two-dimensional data display approaches for UPLC-MS in biomarker discovery and pathway analysis. Anal Chem, 2006, 78(13): 4398–4408

  12. 12

    Holmes E, Cloarec O, Nicholson J K. Probing latent biomarker signatures and in vivo pathway activity in experimental disease states via statistical total correlation spectroscopy (STOCSY) of biofluids: Application to HgCl2 toxicity. J Proteom Res, 2006, 5(6): 1313–1320

  13. 13

    Holmes E, Loo R L, Cloarec O, Coen M, Tang H R, Maibaum E, Bruce S, Chan Q, Elliottm P, Stamler J, Wilson I D, Lindon J C, Nicholson J K. Detection of urinary drug metabolite (Xenometabolome) signatures in molecular epidemiology studies via statistical total correlation (NMR) spectroscopy. Anal Chem, 2007, (79): 2629–2640

  14. 14

    Liu M L, Tang H R, Nicholson J K, Lindon J C. Recovery of underwater resonances by magnetization transferred NMR spectroscopy (RECUR-NMR). J Magn Reson, 2001, 153(1): 133–137

  15. 15

    Liu M L, Tang H R, Nicholson J K, lindon J C. Use of 1H NMR determined diffusion coefficients to characterize lipoprotein fractions in human blood plasma. Magn Reson Chem, 2002, S83–S88

  16. 16

    Brindle J T, Antti H, Holmes E, Tranter G, Nicholson J K, Bettell H W L, Clarke S, Schofield P M, McKilligin E, Mosedale D E, Grainger D J. Rapid and noninvasive diagnosis of the presence and severity of coronary heart disease using 1H-NMR-based metabonomics. Nat Med, 2002, 8(12): 1439–1444

  17. 17

    Athersuch T J, Keun H, Nicholson J K. Quantitative urinalysis of the mercapturic acid conjugates of allyl formate using high resolution NMR spectroscopy. J Pharm Biomed Anal, 2006, 40: 410–416

  18. 18

    Wang Y L, Holmes E, Tang H R, Lindon J C, Sprenger N, Turini M E, Bergonzelli G, Fay L B, Kochhar S, Nicholson J K. Experimental metabonomic model of dietary variation and stress interactions. J Proteome Res, 2006, 5: 1535–1542

  19. 19

    Duarte I F, Stanley E G, Holmes E, Lindon J C, Tang H R, Ferdinand R, McKee C G, Nicholson J K, Vilca-melendze H, Heaton N, Murphy G M. Metabolic assessment of human liver transplants from biopsy samples at the donor and recipient stages using high-resolution magic angle spinning 1H NMR spectroscopy. Anal Chem, 2005, 77(17): 5570–5578

  20. 20

    Wang Y L, Tang H R, Nicholson J K, Hylands P J, Sampson J, Holmes E. A metabonomic strategy for the detection of the metabolic effects of chamomile (Matricaria recutita L.) ingestion. J Agri Food Chem, 2005, 53(2): 191–196

  21. 21

    Wang Y L, Hylands P J, Sampson J, Whitcombe I, Stewart C G, Caiger S, Oru I, Holmes E, Nicholson J K, Metabolomic strategy for the classification and quality control of phytomedicine: A case study of chamomile flower (Matricaria recutita L.). Plant Med, 2004, 70(3): 250–255

  22. 22

    Plumb R S, Johnson K A, Rainville P, Shockcor J P, Williams R, Granger J H, Wilson I D. The detection of phenotypic differences in the metabolic plasma profile of three strains of Zucker rats at 20 weeks of age using ultra-performance liquid chromatography/orthogonal acceleration time-of-flight mass spectrometry. Rapid Commun Mass Spectr, 2006, 20(19): 2800–2806

  23. 23

    Wilson I D, Plumb R, Granger J, Major H, Williams R, Lenz E M. HPLC-MS-based methods for the study of metabonomics. J Chromat Anal Technol Biomed Life Sci, 2005, 817(1): 67–76

  24. 24

    Yin P Y, Zhao X J, Li Q R, Wang J S, Li J H, Xu G W. Metabonomics study of intestinal fistulas based on ultraperformance liquid chromatography coupled with Q-TOF mass spectrometry (UPLC/Q-TOF MS). J Proteom Res, 2006, 5(9): 2135–2143

  25. 25

    Yang J, Zhao X J, Liu X L, Wang C, Gao P, Wang J S, Li L J, Gu J R, Yang S L, Xu G W. High performance liquid chromatography-mass spectrometry for metabonomics: Potential biomarkers for acute deterioration of liver function in chronic hepatitis B. J Proteom Res, 2006, 5(3): 554–561

  26. 26

    Qiu Y, Su M, Liu Y, Chen M, Gu J, Zhang J, Jia W. Application of ethyl chloroformate derivatization for gas chromatography-mass spectrometry based metabonomic profiling. Anal Chim Acta, 2007, 583(2): 277–283

  27. 27

    Tang H R, Wang Y L, Nicholson J K, Lindon J C. Use of relaxation-edited one-dimensional and two dimensional nuclear magnetic resonance spectroscopy to improve detection of small metabolites in blood plasma. Anal Biochem, 2004, 325(2): 260–272

  28. 28

    Wang Y L, Bollard M E, Keun H, Antti H, Beckonert O, Ebbels T M, Lindon J C, Holmes E, Tang H R, Nicholson J K. Spectral editing and pattern recognition methods applied to high-resolution magic-angle spinning 1H nuclear magnetic resonance spectroscopy of liver tissues. Anal Biochem, 2003, 323(1): 26–32

  29. 29

    Martin F P J, Dumas M E, Wang Y L, Legido-Quigley C, Yap I K S, Tang H R, Murphy Z G M, Cloarec O, Lindon J C, Sprenger N, Fay E L B, Kochlar S, Holmes E, Nicholson J K. A top-down systems biology view of microbiome-mammalian metabolic interactions in a mouse model. Mol Systems Biol, 2007, 3: 112

  30. 30

    Yang Y X, Li C L, Nie X, Feng X S, Chen W X, Yue Y, Tang H R, Deng F. Metabonomic studies of human hepatocellular carcinoma using high-resolution magic-angle spinning 1H NMR spectroscopy in conjunction with multivariate data analysis. J Proteom Res, 2007, 6: 2605–2614

  31. 31

    Wang Y L, Tang H R, Holmes E, Lindon J C, Turini M E, Sprenger N, Bergonzelli G, Fay L B, Kochhar S, Nicholson J K. Biochemical characterization of rat intestine development using high-resolution magic-angle-spinning 1H NMR spectroscopy and multivariate data analysis. J Proteom Res, 2005, 4(4): 1324–1329

  32. 32

    Wang Y L, Holmes E, Comelli E M, Fotopoulos G, Dorta G, Tang H R, Rantalainen M, Lindon J C, Corthesy I E, Fay L B, Kochhar D, Nicholson J K. Topographical variation in metabolic signatures of human gastrointestinal biopsies revealed by high-resolution magic-angle spinning 1H NMR spectroscopy. J Proteom Res, 2007: 6(10): 3944–3951

  33. 33

    Bax A, Davis D G, MLEV-17-based two-dimensional homonuclear magnetisation transfer spectroscopy. J Magn Reson, 1985, 65(2): 355–360

  34. 34

    Wang Y L, Holmes E, Nicholson J K, Cloarec O, Chollet J, Tanner M, Singer B H, Utzinger J. Metabonomic investigations in mice infected with Schistosoma mansoni: An approach for biomarker identification. Proc Natl Acad Sci 2004, 101(34): 12676–12681

  35. 35

    Wang Y L, Utzinger J, Xiao S H, Xue J, Nicholson J K, Marcel T S B H, Holmes E. System level metabolic effects of a Schistosoma japonicum infection in the Syrian hamster. Mol Biochem Parasitol, 2006, 146(1): 1–9

  36. 36

    Fan T W M, Metabolite profiling by one-and two-dimensional NMR analysis of complex mixtures. Prog NMR Spectr, 1996, 28: 161–219

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Author information

Correspondence to WeiJin Yang or HuiRu Tang.

Additional information

Supported by the Chinese Academy of Sciences (Grant No: KZCX2-YW-420-21), the National High Technology Research and Development Program of China (Grant No: 2006AA06Z424), and the National Natural Science Foundation of China (Grant Nos: 40590392 and 20575074)

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Yang, W., Wang, Y., Zhou, Q. et al. Analysis of human urine metabolites using SPE and NMR spectroscopy. Sci. China Ser. B-Chem. 51, 218–225 (2008). https://doi.org/10.1007/s11426-008-0031-6

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Keywords

  • urine metabolites
  • NMR spectroscopy
  • solid phase extraction
  • metabonomics/metabolomics