Skip to main content
SpringerLink
Log in

Stereospecific analysis of triacylglycerols rich in long-chain polyunsaturated fatty acids

  • Methods
  • Published:
Lipids

Abstract

Six oils of marine, algal, and microbial origin were analyzed for stereospecific distribution of component fatty acids. The general procedure involved preparation ofsn-1,2-(2,3)-diacylglycerols by partial deacylation with ethylmagnesium bromide or pancreatic lipase, separation of X-1,3- andsn-1,2(2,3)-diacylglycerols by borate thin-layer chromatography, resolution of thesn-1,2- andsn-2,3-enantiomers by chiral phase high-performance liquid chromatography following preparation of dinitrophenylurethane derivatives, and determination of the fatty acid composition by gas chromatography. Unexpected complications arose during a stereospecific analysis of triacylglycerols containing over 33% of either 20∶4 or 22∶6 fatty acids. Thesn-1,2(2,3)-diacylglycerols made up of two long-chain polyunsaturated acids migrated with the X-1,3-diacylglycerols and required separate chiral phase resolution. Furthermore, the enzymatic method yieldedsn-1,2(2,3)-diacylglycerols, overrepresenting the polyenoic species due to their relative resistance to lipolysis, but prolonged digestion yielded correct composition for the 2-monoacylglycerols. The final positional distribution of the fatty acids was established by pooling and normalizing the data from subfractions obtained by norman- and chiral-phase separation of diacylglycerols. The molecular species of X-1,3-,sn-1,2- andsn-2,3-diacylglycerol dinitrophenylurethanes were identified by chiral-phase liquid chromatography/mass spectrometry with electrospray ionization, which demonstrated a preferential association of the paired long-chain acids with thesn-1,2- andsn-2,3-diacylglycerol isomers.

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

Similar content being viewed by others

Abbreviations

DAG:

diacylglycerol

DNPU:

dinitrophenylurethane

ECN:

equivalent carbon number

ELSD:

evaporative light-scattering detection

GC:

gas chromatography

HPLC:

high-performance liquid chromatography

LC/MS:

liquid chromatography/mass spectrometry

MAG:

monoacylglycerols

PN:

partition number

TAG:

triacylglycerol

TLC:

thin-layer chromatography

References

  1. Ransac, S., Rogalska, E., Gargouri, Y., Deveer, A., Paltauf, F., de Haas, G.H., and Verger, R. (1990) Stereoselectivity of Lipases. I. Hydrolysis of Enantiomeric Glyceride Analogues by Gastric and Pancreatic Lipases, a Kinetic Study Using the Monomolecular Film Technique,J. Biol. Chem. 265, 20263–20270.

    PubMed  CAS  Google Scholar 

  2. Rogalska, E., Ransac, S., and Verger, R. (1990) Stereoselectivity of Lipases. II. Stereoselective Hydrolysis of Triglycerides by Gastric and Pancreatic Lipases,J. Biol. Chem. 265, 20271–20276.

    PubMed  CAS  Google Scholar 

  3. Nilsson, A., Landin, B., and Schotz, M.C. (1987) Hydrolysis of Chylomicron Arachidonate and Linoleate Ester Bonds by Lipoprotein and Hepatic Lipase.J. Lipid Res. 28, 510–517.

    PubMed  CAS  Google Scholar 

  4. Clark, B., and Huebscher, G. (1960) Biosynthesis of Glycerides in Subcellular Fractions of Intestinal Mucosa,Biochim. Biophys. Acta 152, 479–494.

    Google Scholar 

  5. Yang, L.-Y., and Kuksis, A. (1991) Apparent Convergence (at 2-monoacylglycerol level) of Phosphatidic Acid and 2-Monoacylglycerol Pathways of Synthesis of Chylomicron Triacylglycerols,J. Lipid Res. 32, 1173–1186.

    PubMed  CAS  Google Scholar 

  6. Carlson, W.E., Werkman, S.H. Peeples, J.M., Cooke, R.J., and Tolley, E.A. (1993) Arachidonic Acid Status Correlates with First Year Growth in Preterm Infants,Proc. Natl. Acad. Sci. 90 1072–1077.

    Google Scholar 

  7. Crastes de Paulet, A., Babib, F., Billeaud, C., Sarda, P., and Mendy, F. (1994) Effets Biologiques chez le Premature, d'une Supplementation des Formules Lactees en Acide Alpha-Linolenique: une etude Multicentrique,Bull. Acad. Natl. Med. 178, 267–273.

    PubMed  CAS  Google Scholar 

  8. Kyle, D.J., and Rattledge, C. (eds.)Industrial Applications of Single Cell Oils AOCS Press, Champaign, 1992.

    Google Scholar 

  9. Brockerhoff, H., Hoyle, R.J., Hwang, P.C., and Litchfield, C. (1968) Positional Distribution of Fatty Acids in Depot Triglycerides of Aquatic Animals,Lipids 3, 24–29.

    CAS  PubMed  Google Scholar 

  10. Yang, L.-Y., Kuksis, A., and Myher, J.J. (1991) Surface Lipids of Chylomicrons from Rats Fed Glyceryl and Alkyl Esters of Mixed Fatty Acids: Major Components,Lipids 26, 806–818.

    PubMed  CAS  Google Scholar 

  11. Itabashi, Y., Kuksis, A., Marai, L., and Takagi, T. (1990) Chiral Phase High Performance Liquid Chromatographic Resolution of Diacylglycerol Moieties of Natural Triacylglycerols on R-+-1-(1-naphthylethylamine),J. Lipid Res. 31, 1711–1717.

    PubMed  CAS  Google Scholar 

  12. Myher, J.J., and Kuksis, A. (1982) Resolution of Natural Diacylglycerols by Gas-Liquid Chromatography on Polar Capillary Columns,Can. J. Biochem., 60, 638–650.

    Article  PubMed  CAS  Google Scholar 

  13. Thomas, A.E., Scharoun J.E., and Ralston, H. (1965) Quantitative Estimation of Isomeric Monoglycerides by Thin-Layer Chromatography,J. Am. Oil Chem. Soc. 42, 789–792.

    CAS  Google Scholar 

  14. Myher, J.J., Kuksis, A., and Yang, L.-Y. (1990) Stereospecific Analysis of Menhaden Oil Triacylglycerols and Resolution of Complex Polyunsaturated Diacylglycerols by Gas-Liquid Chromatography on Polar Capillary Columns.Biochem. Cell Biol. 68, 336–344.

    Article  CAS  Google Scholar 

  15. Marai, L., Kuksis, A., and Myher, J.J. (1994) Reversed-Phase Liquid Chromatography-Mass Spectrometry of the Uncommon Triacylglycerol Structures Generated by Randomization of Butter Oil,J. Chromatogr. A. 672, 87–99.

    Article  CAS  Google Scholar 

  16. Myher, J.J., Kuksis, A., Ravandi, A., and Cocks, N. (1994) Identification of Polyoxyglycerophospholipids by Normal Phase LC/MS with Electrospray,INFORM 5, 478–479.

    Google Scholar 

  17. Martin, J.-C., Bougnoux, P. Antoine, J.-M., Lanson, M., and Couet, C. (1993) Triacylglycerol Structure of Human Colostrum and Mature Milk.Lipids 28, 637–643.

    PubMed  CAS  Google Scholar 

  18. Innis, S.M. (1992) n-3 Fatty Acid Requirements of the New-born,Lipids 27, 879–885.

    PubMed  CAS  Google Scholar 

  19. Werkman, S.H., and Carlson, S.E. (1995) Preterm Infants Fed Formula Supplemented with Marine Oil DHA Compared with Controls Fed Only Linolenic Acid Have a Shorter Look Duration,INFORM 6, 530, Abs. No. B.

  20. Yang, L.-Y., Kuksis, A., and Myher, J.J. (1990) Lipolysis of Menhaden Oil Triacylglycerols and the Corresponding Fatty Acid Alkyl Esters by Pancreatic Lipasein vitro: A Reexamination,J. Lipid Res. 31, 192–204.

    Google Scholar 

  21. Bottino, R., Vandenburg, G.A., and Reiser, R. (1967) Resistance of Certain Long-Chain Polyunsaturated Fatty Acids of Marine Oils to Pancreatic Lipase Hydrolysis,Lipids 2, 489–493.

    CAS  PubMed  Google Scholar 

  22. Miyashita, K., Frankel, E.N., Neff, W.E., and Awl, R.A. (1990) Autoxidation of Polyunsaturated Triacylglycerols. III. Synthetic Triacylglycerols Containing Linoleate and Linolenate,Lipids 25, 48–53.

    CAS  Google Scholar 

  23. Wada, S., and Koizumi, C. (1983) Influence of the Position of Unsaturated Fatty Acid Esterified Glycerol on the Oxidation Rate of Triglyceride.J. Am. Oil Chem. Soc. 60, 1105–1109.

    CAS  Google Scholar 

  24. Raghuveer, G., and Hammond, E.G. (1967) The Influence of Glyceride Structure on the Rate of Autoxidation,J. Am. Oil Chem. Soc. 44, 239–243.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Banting and Best Department of Medical Research, University of Toronto, Toronto, Canada

    J. J. Myher, A. Kuksis & K. Geher

  2. Mead Johnson Research Center, Evansville, Indiana

    P. W. Park & D. A. Diersen-Schade

Authors
  1. J. J. Myher
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Kuksis
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Geher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. W. Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D. A. Diersen-Schade
    View author publications

    You can also search for this author in PubMed Google Scholar

About this article

Cite this article

Myher, J.J., Kuksis, A., Geher, K. et al. Stereospecific analysis of triacylglycerols rich in long-chain polyunsaturated fatty acids. Lipids 31, 207–215 (1996). https://doi.org/10.1007/BF02522622

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02522622

Keywords

Search

Navigation