Skip to main content
SpringerLink
Log in

High-Resolution NMR Spectroscopy: An Alternative Fast Tool for Qualitative and Quantitative Analysis of Diacylglycerol (DAG) Oil

  • Original Article
  • Published:
Journal of the American Oil Chemists' Society

Abstract

Multinuclear (1H, 13C, 31P) and multidimensional NMR spectroscopy was employed for the analysis of diacylglycerol (DAG) oil and the quantification of its acylglycerols and acyl chains composition. A number of gradient selected two dimensional NMR techniques (TOCSY, HSQC-DEPT, HSQC-TOCSY, and HMBC) facilitated the assignment of the complex one dimensional 1H- and 13C-NMR spectra. In several cases, the aforementioned 2D-NMR techniques offered solid proof of earlier assignments based on chemical shift changes induced by the presence and relative positions of double bonds within the acyl chains. Integration of the appropriate signals in the NMR spectra of the three nuclei allowed the determination of DAG oil composition, which was found to be within the limits accepted for this oil, namely 1-monoacylglycerols 0.40–0.60%; 2-monoacylglycerols 0.40–0.50%; 1,3-diacylglycerols 57–62%; 1,2-diacylglycerols 28–32%; triacylglycerols 9–11%; saturated fatty acids 3–5%; oleic acid 37–45%; linoleic acid 49–53%; and linolenic acid 5–6.5%; tocopherols 0.24–0.27%. The compositional results obtained from the NMR spectra of the three nuclei were compared and discussed in terms of repeatability and ease of performance.

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

References

  1. Katsuragi K, Yasukawa T, Matsuo N, Flickinger B, Tokimitsu I, Matlock M (2008) Diacylglycerol oil, 2nd edn. AOCS Press, Urbana

    Google Scholar 

  2. Sikorski D (2004) Application of diacylglycerol oil in baked goods, nutritional beverages/bars, sauces, and gravies. In: Katsuragi Y, Yasukawa T, Matsuo N, Flickinger B, Tokimitsu I, Matlock M (eds) AOCS Press, Champaign

  3. Lo SK, Tan CP, Long K, Suria M, Yusoff A, Lai OM (2008) Diacylglycerol oil-properties, processes and products: a review. Food Biopr Technol 1:223–233

    Article  Google Scholar 

  4. Lo SK, Baharin BS, Tan CP, Laia OM (2004) Lipase-catalysed production and chemical composition of diacylglycerols from soybean oil deodoriser distillate. Eur J Lipid Sci Technol 106:218–224

    Article  CAS  Google Scholar 

  5. AOCS (2003) Determination of mono- and diglycerides by capillary gas chromatography, Method Cd 11b-91. In: Official Methods and Recommended Practices of the American Oil Chemists, 5th edn, AOCS press, Champaign

  6. Watanaber T, Yamagushi H, Yamada N, Lee I (2004) Manufacturing process of diacylglycerol oil. In: Katsuragi Y, Yasukawa T, Matsuo N, Flickinger B, Tokimitsu I, Matlock M (eds). AOCS Press, Champaign

  7. Sacchi R, Addeo F, Paolillo L (1997) 1H and 13C NMR of virgin oil. An overview. Magn Reson Chem 35:5133–5145

    Article  Google Scholar 

  8. Vlahov G (1999) Application of NMR to the study of olive oils. Progr Nucl Magn Reson Spectrosc 35:341–357

    Article  CAS  Google Scholar 

  9. Mannina L, Luchinat C, Emanuele MC, Segre A (1999) Acyl positional distribution of glycerol tri-esters in vegetable oils: a 13C NMR study. Chem Phys Lipids 103:47–55

    Article  CAS  Google Scholar 

  10. Spyros A, Dais P (2000) Application of 31P NMR spectroscopy in food analysis. I. Quantitative determination of the mono- and diglyceride composition of olive oils. J Agric Food Chem 48:802–805

    Article  CAS  Google Scholar 

  11. Dais P, Spyros A (2007) 31P NMR spectroscopy in the quality control and authentication of virgin olive oil. An account of recent results. Magn Reson Chem 45:367–377

    Article  CAS  Google Scholar 

  12. Hatzakis E, Dagounakis G, Agiomyrgianaki A, Dais P (2010) A facile NMR method for the quantification of total, free and esterified sterols in virgin olive oil. Food Chem 122:346–352

    Article  CAS  Google Scholar 

  13. Zwierzak A (1967) Cyclic organophosphorus compounds. I. Synthesis and infrared spectral studies of cyclic hydrogen phosphites and thiophosphites. Can J Chem 45:2501–2512

    Article  CAS  Google Scholar 

  14. Lyerla JR, Levy, GC (1974) Carbon-13 nuclear spin relaxation. In: Levy GC (ed) Topics in carbon-13 NMR spectroscopy. Wiley, New York, Chapter 3, pp 100–101

  15. Freeman R, Hill HDW, Kaptein R (1976) Proton-decoupled NMR. Spectra of carbon-13 with the nuclear Overhauser effect suppressed. J Magn Reson 7:327–329

    Google Scholar 

  16. Berger S, Braun S (2004) 200 and more experiments: a practical course. Wiley VCH, Weinheim

    Google Scholar 

  17. Parella T, Sanchez-Ferrando F, Virgili A (1997) Improved sensitivity in gradient-based 1D and 2D multiplicity edited HSQC experiments. J Magn Reson 126:274–277

    Article  CAS  Google Scholar 

  18. Sacchi R, Patumi M, Fontanazza G, Barone P, Fiordiponti P, Mannina L, Rossi E, Segre A (1996) A high field 1H nuclear magnetic resonance study of the minor components in virgin olive oils. J Am Oil Chem Soc 73:747–758

    Article  CAS  Google Scholar 

  19. Guillén MD, Ruiz A (2003) 1H nuclear magnetic resonance as a fast tool for determining the composition of acyl chains in acylglycerol mixtures. Eur J Lipid Sci Technol 105:502–505

    Article  Google Scholar 

  20. Vigli G, Philippidis A, Spyros A, Dais P (2003) Classification of edible oils by employing 31P and 1H NMR Spectroscopy in combination with multivariate statistical analysis. A proposal for detection of seed oils adulteration in virgin olive oils. J Agric Food Chem 51:5715–5722

    Article  CAS  Google Scholar 

  21. Knothe G, Kenar JA (2004) Determination of the fatty acid profile by 1H-NMR spectroscopy. Eur J Lipid Sci Technol 106:88–96

    Article  CAS  Google Scholar 

  22. Rosati O, Albrizio S, Montesano D, Riccieri R, Cossignani L, Curini M, Simonetti S, Rastrelli L, Damiani P (2007) HPLC separation and NMR structural elucidation of sn-1,2, 2,3, and 1,3-diacylglycerols from olive oil as naphthylethylurethane derivatives. J Agric Food Chem 55:191–196

    Article  CAS  Google Scholar 

  23. Sacchi R, Paolillo L, Giudicianni I, Addeo (1991) Rapid 1H-NMR determination of 1,2 and 1,3 diglycerides in virgin olive oils. Ital J Food Sci 4:245–262

    Google Scholar 

  24. Frost DJ, Barzilay J (1971) Proton magnetic resonance identification of nonconjugated cis-unsaturated fatty acids and esters. Anal Chem 43:1316–1318

    Article  CAS  Google Scholar 

  25. Gunstone FD (1991) 13C NMR studies of mono-, di-, and tri-acylglycerols leading to qualitative and semiquantitative information about mixtures of these glycerol esters. Chem Phys Lipids 58:219–224

    Article  CAS  Google Scholar 

  26. Gunstone FD (1993) High resolution 13C NMR spectroscopy of lipids. In: Christie WW (ed) Advances in lipid methodology, 2nd edn. The Oily Press, Dundee, pp 1–68

    Google Scholar 

  27. Wollemberg KF (1990) Quantitative high resolution 13C nuclear magnetic resonance of the olefinic and carbonyl carbons of edible vegetable oils. J Am Oil Chem Soc 67:487–494

    Article  Google Scholar 

  28. Lie Ken Jie MSF, Mustafa J (1997) High-resolution nuclear magnetic resonance spectroscopy—applications to fatty acids and triacylglycerols. Lipids 32:1019–1034

    Article  Google Scholar 

  29. Vlahov G, Schiavone C, Simone N (2001) Quantitative 13C NMR method using DEPT pulse sequence for the determination of the geographical origin (DOP) of olive oils. Magn Reson Chem 39:689–695

    Article  CAS  Google Scholar 

  30. Alenamy LB (2002) Using simple 13C linewidth and relaxation measurements to make detailed chemical shift assignments in triacylglycerols and related compounds. Chem Phys Lipids 120:33–44

    Google Scholar 

  31. Zamora R, Gómez G, Hidalgo FJ (2002) Classification of vegetable oils by high-resolution 13C NMR spectroscopy using chromatographically obtained oil fractions. J Am Oil Chem Soc 79:267–272

    Article  CAS  Google Scholar 

  32. Simonova S, Ivanova G, Spassov SL (2003) Alternative NMR method for quantitative determination of acyl positional distribution in triacylglycerols and related compounds. Chem Phys Lipids 126:167–176

    Article  Google Scholar 

  33. Vlahov G, Giuliani AA, Del Re P (2010) 13C NMR spectroscopy for determining the acylglycerol positional composition of lampante olive oils. Chemical shift assignments and their dependence on sample concentration. Anal Methods 2:916–923

    Article  CAS  Google Scholar 

  34. Mattson FH, Volpenhein RA (1963) The specific distribution of unsaturated fatty acids in the triglycerides of plants. J Lipid Res 4:392–396

    CAS  Google Scholar 

  35. Vlahov G (2005) 13C nuclear magnetic resonance spectroscopy to check 1,3-random, 2-random pattern for fatty acid distribution in olive oil triacylglycerols. Spectroscopy 19:109–117

    CAS  Google Scholar 

  36. Willker W, Leibfritz D (1998) Assignment of mono- and polyunsaturated fatty acids in lipids of tissues and body fluids. Magn Reson Chem 36:S79–S84

    Article  CAS  Google Scholar 

  37. Vatele JM, Fenet B, Eynard T (1998) Complete 13C assignment and structural elucidation of n-3 polyunsaturated fatty acids by the use of a new 2D NMR technique SAPHIR-HSQC. Chem Phys Lipids 94:239–250

    Article  CAS  Google Scholar 

  38. Pauli GF, Jaki BU, Lankin DC (2005) Quantitative 1H NMR: development and potential of a method for natural products analysis. J Nat Prod 68:133–149

    Article  CAS  Google Scholar 

  39. Shoolery JN (1976) Some quantitative applications of 13C NMR spectroscopy. Progr NMR Spectrosc 11:79–93

    Article  Google Scholar 

  40. Vlahov G (1998) Regiospecific analysis of natural mixtures of triglycerides using quantitative 13C nuclear magnetic resonance of acyl chain carbons. Magn Reson Chem 36:359–362

    Article  CAS  Google Scholar 

  41. Dais P (1989) Rotational dynamics of flexible-chain molecules. 13C NMR relaxation study of hydrocarbon chains attached to a heavy anchor. Magn Reson Chem 27:61–67

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The 600 MHz NMR spectrometer used in this study was purchased with the CHE-0821552 grant from the NSF.

Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of North Carolina Wilmington, 601 S. College Rd., 253 Dobo Hall, Wilmington, NC, 28403, USA

    Emmanuel Hatzakis

  2. NMR Laboratory, Department of Chemistry, University of Crete, P.O. Box 2208, Voutes Campus, 710 03, Heraklion, Crete, Greece

    Alexia Agiomyrgianaki, Sarantos Kostidis & Photis Dais

Authors
  1. Emmanuel Hatzakis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexia Agiomyrgianaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sarantos Kostidis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Photis Dais
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Photis Dais.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 254 kb)

About this article

Cite this article

Hatzakis, E., Agiomyrgianaki, A., Kostidis, S. et al. High-Resolution NMR Spectroscopy: An Alternative Fast Tool for Qualitative and Quantitative Analysis of Diacylglycerol (DAG) Oil. J Am Oil Chem Soc 88, 1695–1708 (2011). https://doi.org/10.1007/s11746-011-1848-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11746-011-1848-2

Keywords

Search

Navigation