Skip to main content
SpringerLink
Log in

Improved quantification from 1H-NMR spectra using reduced repetition times

  • Original Article
  • Published:
Metabolomics Aims and scope Submit manuscript

Abstract

A method of choosing the correction factor based on Bloch equation for the quantitative estimation of metabolite from 1H NMR spectra recorded with reduced recycle delay is prescribed. The procedure reduces the experimental time without substantially compromising the accuracy of quantitative estimation. It is based on choosing the correction factors, which depend on T1 and T2 of the metabolite and recycle delay used for recording the spectra. It is validated by studying a mixture of amino acids with known concentration of constituents and human serum sample and it provides accuracy of quantitative estimation to 95–96%.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

CPMG:

Carr–Purcell–Meiboom–Gill

FID:

Free induction decay

qNMR:

Quantitative NMR

TSP:

Trimethyl silane propionic acid sodium salt

References

  • Akoka, S., Barantin, L., & Trierweiler, M. (1999). Concentration measurement by proton NMR using the ERETIC method. Analytical Chemistry, 71, 2554–2557. doi:10.1021/ac981422i.

    Article  CAS  Google Scholar 

  • Bala, L., Ghoshal, U. C., Ghoshal, U., et al. (2006). Malabsorption syndrome with and without small intestinal bacterial overgrowth: A study on upper-gut aspirate using 1H NMR spectroscopy. Magnetic Resonance in Medicine, 56, 738–744. doi:10.1002/mrm.21041.

    Article  PubMed  Google Scholar 

  • Bala, L., Gowda Nagana, G. A., Ghoshal, U. C., Misra, A., Bhandari, M., & Khetrapal, C. L. (2004). 1H NMR spectroscopic method for diagnosis of malabsorption syndrome: A pilot study. NMR in Biomedicine, 17, 69–75. doi:10.1002/nbm.866.

    Article  PubMed  CAS  Google Scholar 

  • Bell, J. D., Brown Judith, C. C., & Sadler, P. J. (1989). NMR studies of body fluids. NMR in Biomedicine, 2, 246–256. doi:10.1002/nbm.1940020513.

    Article  PubMed  CAS  Google Scholar 

  • Bell, J. D., Lee, J. A., Lee, H. A., Sadler, P. J., Wilkie, D. R., & Woodham, R. H. (1991). Nuclear magnetic resonance studies of blood plasma and urine from subjects with chronic renal failure: Identification of trimethylamine-N-oxide. Biochimica et Biophysica Acta, 1096, 101–107.

    PubMed  CAS  Google Scholar 

  • Bell, J. D., & Sadler, P. J. (1999). Body fluids. In Encyclopedia of magnetic resonance (vol. 2, pp. 989–1001). New York: Wiley.

  • Bloch, F. (1946). Nuclear induction. Physical Review, 70, 460–474. doi:10.1103/PhysRev.70.460.

    Article  CAS  Google Scholar 

  • Carr, H. Y., & Purcell, E. M. (1954). Effect on diffusion on free precision in nuclear magnetic resonance experiments. Physical Review, 94, 630–638. doi:10.1103/PhysRev.94.630.

    Article  CAS  Google Scholar 

  • Caytana, E., Remaud, G. S., Tenailleaua, E., & Akoka, S. (2007). Precise and accurate quantitative 13C NMR with reduced experimental time. Talanta, 71(3), 1016–1021. doi:10.1016/j.talanta.2006.05.075.

    Article  Google Scholar 

  • Daviesa, S. E. C., Woolf, D. A., Chalmers, R. A., Raftera, J. E. M., & Iles, R. A. (1992). Proton nmr studies of betaine excretion in the human neonate: consequences for choline and methyl group supply. The Journal of Nutritional Biochemistry, 3(10), 523–530. doi:10.1016/0955-2863(92)90074-S.

    Article  Google Scholar 

  • Daykin, C. A., Foxall, P. J. D., Connor, S. C., Lindon, J. C., & Nicholson, J. K. (2002). The comparison of plasma deproteinization methods for the detection of low-molecular-weight metabolites by 1H nuclear magnetic resonance spectroscopy. Analytical Biochemistry, 304, 220–230. doi:10.1006/abio.2002.5637.

    Article  PubMed  CAS  Google Scholar 

  • de Graaf, R. A., & Behar, K. L. (2003). Quantitative 1H NMR spectroscopy of blood plasma metabolites. Analytical Chemistry, 75(9), 2100–2104. doi:10.1021/ac020782+.

    Article  PubMed  Google Scholar 

  • Evilia, R. F. (2001). Quantitative NMR spectroscopy. Analytical Letters, 34(13), 2227–2236. doi:10.1081/AL-100107290.

    Article  CAS  Google Scholar 

  • Gang, S., Kautz, R., Shiqi, P., Guohui, C., & Giese, R. (2007). Calibration by NMR for quantitative analysis: P-Toluenesulfonic acid as a reference substance. Journal of Chromatography A, 1138, 305–308. doi:10.1016/j.chroma.2006.10.010.

    Article  Google Scholar 

  • Griffiths, L., & Irving, A. M. (1998). Assay by nuclear magnetic resonance spectroscopy: Quantification limits. Analyst (London), 123, 1061–1068. doi:10.1039/a800625c.

    Article  CAS  Google Scholar 

  • Hahn, E. L. (1950). Spin echoes. Physical Review, 80, 580–594. doi:10.1103/PhysRev.80.580.

    Article  Google Scholar 

  • Henderson, T. J. (2002). Quantitative NMR spectroscopy using coaxial inserts containing a reference standard: Purity determinations for Military nerve agents. Analytical Chemistry, 74, 191–198. doi:10.1021/ac010809+.

    Article  PubMed  CAS  Google Scholar 

  • Kriat, M., Gouny, S. C., Dury, J. V., Sciaky, M., Viout, P., & Cozzone, J. P. (1992). Quantitation of metabolites in human blood serum by proton magnetic resonance. A comparative study of the use of Formate and TSP as concentration standards. NMR in Biomedicine, 5, 179–184. doi:10.1002/nbm.1940050404.

    Article  PubMed  CAS  Google Scholar 

  • Larive, C. K., Jayawickrama, D., & Orfi, L. (1997). Quantitative analysis of peptides with NMR spectroscopy. Applied Spectroscopy, 51, 1531–1536. doi:10.1366/0003702971939055.

    Article  CAS  Google Scholar 

  • Lindon, J. C., Holmes, E., Bollard, M. E., Stanley, E. G., & Nicholson, J. K. (2004). Metabonomics technologies and their applications in physiological monitoring, drug safety assessment and disease diagnosis. Biomarkers, 9(1), 1–31.

    Article  PubMed  CAS  Google Scholar 

  • Lindon, J. C., Nicholson, J. K., Holmes, E., & Everett, J. R. (2000). Metabonomics: Metabolic processes studied by NMR spectroscopy of biofluids. Concepts in Magnetic Resonance, 12(5), 289–320. doi:10.1002/1099-0534(2000)12:5<289::AID-CMR3>3.0.CO;2-W.

    Article  CAS  Google Scholar 

  • Lucas, L. H., Larive, C. K., Wilkinson, P. S., & Huhn, S. (2005). Progress toward automated metabolic profiling of human serum: Comparison of CPMG and gradient-filtered NMR analytical methods. Journal of Pharmaceutical and Biomedical Analysis, 39, 156–163. doi:10.1016/j.jpba.2004.09.060.

    Article  PubMed  CAS  Google Scholar 

  • Malz, F., & Jancke, H. (2005). Validation of quantitative NMR. Journal of Pharmaceutical and Biomedical Analysis, 38(5), 813–823. doi:10.1016/j.jpba.2005.01.043.

    Article  PubMed  CAS  Google Scholar 

  • Meiboom, S., & Gill, D. (1958). Modified spin echo method for measuring nuclear magnetic relaxation times. The Review of Scientific Instruments, 29, 688–691. doi:10.1063/1.1716296.

    Article  CAS  Google Scholar 

  • Nishijima, T., Nishina, M., & Fujiwara, K. (1997). Measurement of lactate levels in serum and bile using proton nuclear magnetic resonance in patients with hepatobiliary diseases: Its utility in detection of malignancies. Japanese Journal of Clinical Oncology, 27(1), 13–17. doi:10.1093/jjco/27.1.13.

    Article  PubMed  CAS  Google Scholar 

  • Pauli, G. F., Jaki, B. U., & Lankin, D. C. (2006). Quantitative 1H NMR: Development and potential of a method for natural products analysis. Journal of Natural Products, 68, 133–149. doi:10.1021/np0497301.

    Article  Google Scholar 

  • Saude, E. J., Slupsky, C. M., & Sykes, B. D. (2006). Optimization of NMR analysis of biological fluids for quantitative accuracy. Metabolomics, 2, 113–123. doi:10.1007/s11306-006-0023-5.

    Article  CAS  Google Scholar 

  • Saxena, V., Gupta, A., Gupta Nagana, G. A., Saxena, R., Yachha, S. K., & Khetrapal, C. L. (2006). 1H NMR spectroscopy for the prediction of therapeutic outcome in patients with fulminant hepatic failure. NMR in Biomedicine, 19(5), 521–526. doi:10.1002/nbm.1034.

    Article  PubMed  CAS  Google Scholar 

  • Shiyan, F., Choy, W. Y., Lam, S. L., & Au-Yeung, S. C. F. (1992). Quantitative determination of glucose in blood plasma and in fruit juices by combined WATR-CPMG 1H NMR spectroscopy. Analytical Chemistry, 64, 2570–2574. doi:10.1021/ac00045a018.

    Article  Google Scholar 

  • Silvestre, V., Goupry, S., Trierweiler, M., Robins, R., & Akoka, S. (2001). Determination of substrate and product concentration in lactic acid bacterial fermentations by proton NMR using the ERETIC method. Analytical Chemistry, 73, 1862–1868. doi:10.1021/ac0013204.

    Article  PubMed  CAS  Google Scholar 

  • Singh, H. K., Yachha, S. K., Saxena, R., et al. (2006). New dimension of 1H-NMR spectroscopy in assessment of liver graft dysfunction. NMR in Biomedicine, 16(4), 185–188. doi:10.1002/nbm.829.

    Article  Google Scholar 

  • Subramanian, A., Gupta, A., Saxena, S., et al. (2005). Proton MR CSF analysis and a new software as predictors for the differentiation of meningitis in children. NMR in Biomedicine, 18, 213–225. doi:10.1002/nbm.944.

    Article  PubMed  CAS  Google Scholar 

  • Van, Q. N., Chmurny, G. N., & Veenstra, T. D. (2003). The depletion of protein signals in metabonomics analysis with the WET-CPMG pulse sequence. Biochemical and Biophysical Research Communications, 301, 952–959. doi:10.1016/S0006-291X(03)00079-2.

    Article  PubMed  CAS  Google Scholar 

  • Viant, M., Rosenblum, E., & Tjeerdema, R. (2003). NMR-based metabolomics: A powerful approach for characterizing the effects of environmental stressors on organism health. Environmental Science & Technology, 37, 4982–4989. doi:10.1021/es034281x.

    Article  CAS  Google Scholar 

  • Voet, D., & Voet, J. G. (1990). Biochemistry. Canada: Wiley.

    Google Scholar 

  • Wevers, R. A., Engelke, U., & Heerschap, A. (1994). High-resolution 1HNMR spectroscopy of blood plasma for metabolic studies. Clinical Chemistry, 40(7), 1245–1250.

    PubMed  CAS  Google Scholar 

  • Yamaguchi, S., Koda, N., & Yamamoto, H. (1989). Analysis for homogentisic acid by NMR spectrometry, to aid diagnosis of Alkaptonurla. Clinical Chemistry, 35(8), 1806–1807.

    PubMed  CAS  Google Scholar 

Download references

Acknowledgement

The authors gratefully acknowledge the financial support from the Department of Science and Technology, Government of India.

Author information

Authors and Affiliations

  1. Centre of Biomedical Magnetic Resonance, Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS) Campus, Raibarelli Road, Lucknow, 226014, Uttar Pradesh, India

    Santosh Kumar Bharti, Neeraj Sinha, Sudhir Kumar Mandal, Raja Roy & Chunni Lal Khetrapal

  2. Bruker India Scientific Pvt. Ltd., Lucknow, Uttar Pradesh, India

    Bhawani Shankar Joshi

Authors
  1. Santosh Kumar Bharti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Neeraj Sinha
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bhawani Shankar Joshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sudhir Kumar Mandal
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Raja Roy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chunni Lal Khetrapal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chunni Lal Khetrapal.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bharti, S.K., Sinha, N., Joshi, B.S. et al. Improved quantification from 1H-NMR spectra using reduced repetition times. Metabolomics 4, 367–376 (2008). https://doi.org/10.1007/s11306-008-0130-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11306-008-0130-6

Keywords

Search

Navigation