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Synthesis of acetyl, docosahexaenoyl-glycerophosphocholine and its characterization using nuclear magnetic resonance

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Lipids

Abstract

Docosahexaenoic acid (DHA) circulates in mammals in lipoproteins and bound to serum albumin as a nonesterified fatty acid as well as esterified in lysophosphatidylcholine (lysoPC). 1-Lyso,2-DHA-glycerophosphocholine (GPC) is an unstable isomer because of a primary alcohol at the sn-1 position. To keep DHA at the sn-2 position of lysoPC, its usual position for the corresponding lysoPC to be acylated into PC in tissues, we synthesized 1-acetyl,2-DHA-GPC and confirmed its structure by use of nuclear magnetic resonance (NMR) spectroscopy in comparison with its positional isomer, 1-DHA,2-acetyl-GPC. 1-Lyso,2-DHA-GPC was prepared from 1-stearoyl,2-DHA-GPC by enzymatic hydrolysis and purified by high-performance liquid chromatography. The isomerization of 1-lyso,2-DHA-GPC into 1-DHA,2-lyso-GPC was obtained by keeping the former overnight at room temperature under nitrogen. Both lysoPC isomers were acetylated by acetic anhydride into 1-acetyl,2-DHA-GPC and 1-DHA,2-acetyl-GPC, respectively, and the resulting phospholipids were fully characterized by NMR. In particular, the 1,2 substitution pattern of the acetyl and DHA chains could be easily detected by 2D heteronuclear multibond correlation. We conclude that 1-acetyl,2-DHA-GPC might be considered as a stable form of 1-lyso,2-DHA-GPC for its delivery to tissues, if the latter exhibits acetyl hydrolase activity.

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Abbreviations

COSY:

correlation spectroscopy

DHA:

docosahexaenoic acid

GPC:

glycerophosphocholine

HPLC:

high-performance liquid chromatography

HMBC:

heteronuclear multibond correlation

HSQC:

heteronuclear single quantum coherence

lysoPC:

lysophosphatidylcholine

NMR:

nuclear magnetic resonance

PC:

phosphatidylcholine

TLC:

thin-layer chromatography

References

  1. Bang, H.O., Dyerberg, J., and Hjorne, N. (1976) The Composition of Food Consumed by Greenland Eskimos, Acta Med. Scand. 200, 69–73.

    Article  PubMed  CAS  Google Scholar 

  2. Leaf, A. (1990) Cardiovascular Effects of Fish Oils. Beyond the Platelet, Circulation 82, 624–628.

    PubMed  CAS  Google Scholar 

  3. Anderson, R.E. (1970) Lipids of the Ocular Tissues. IV. A Compoarison of the Phospholipids from the Retina of Six Mammalian Species, Exp. Eye Res. 10, 339–344.

    Article  PubMed  CAS  Google Scholar 

  4. Salem, N., Jr., Kim, H.Y., and Yergey, J.A. (1986) Docosahexaenoic Acid Membrane Function and Metabolism, in Health Effects of Polyunsaturated Fatty Acid in Seafoods (Simopoulos, A.P., Kifer, R.P., and Martin, R., eds.), pp. 263–317, Academic Press, Orlando.

    Google Scholar 

  5. Neuringer, M., Connor, W.E., Lin, D.S., Barstad, L., and Luck, S.J. (1986) Biochemical and Functional Effects of Prenatal and Postnatal Omega-3 Fatty Acid Deficiency on Retina and Brain in Rhesus Monkeys, Proc. Natl. Acad. Sci. USA 83, 285–294.

    Article  Google Scholar 

  6. Makrides, M., Neumann, M.A., and Gibson, R.A. (1996) Is Dietary Docosahexaenoic Acid Essential for Term Infants? Lipids 31, 115–119.

    Article  PubMed  CAS  Google Scholar 

  7. Werkman, S.H., and Carlson S.E. (1996) A Randomized Trial of Visual Attention of Preterm Infants Fed Docosahexanoic Acid Until Nine Months, Lipids 31, 91–97.

    Article  PubMed  CAS  Google Scholar 

  8. Croset, M., Brossard, N., Pachiaudi, C., Normand, S., Lecerf, J., Chirouze, V., Riou, J.P., Tayot, J.L., and Lagarde, M. (1996) In Vivo Compartmental Metabolism of 13C Docosahexaenoic Acid, Studied by Gas Chromatography-Combustion Isotope Ratio Mass Spectrophotometry, Lipids 31, 109–114.

    Google Scholar 

  9. Brossard, N., Croset, M., Lecerf, J., Pachaudi, C., Normand, S., Chirouze, V., Macovschi, O., Riou, J.P., Tayot, J.L., and Lagarde, M. (1996) Metabolic Fate of an Oral Dose of (13C) Docosahexaenoic Acid Triglyceride in the Rat, Am. J. Physiol. 270, R846-R854.

    PubMed  CAS  Google Scholar 

  10. Lemaitre-Delaunay, D., Pachiaudi, C., Laville, M., Pousin, J., Armstrong, M., and Lagarde, M. (1999) Blood Compartmental Metabolism of Docosahexaenoic Acid (DHA) in Humans After Ingestion of a Single Dose of (13C)-Phosphatidylcholine Containing DHA, J. Lipid Res. 40, 1867–1874.

    PubMed  CAS  Google Scholar 

  11. Innis, S.M., (1992) Plasma and Red Blood Cell Fatty Acid Values as Indexes of Essential Fatty Acid in the Developing Organs of Infants Fed with Milk or Formulas, J. Pediatr. 120, 578–586.

    Google Scholar 

  12. Thiès, F., Pillon, C., Molière, P., Lagarde, M., and Lecerf, J. (1994) Preferential Incorporation of sn-2 lysoPC DHA Over Unesterified DHA in the Young Rat Brain, Am. J. Physiol. 267, R1273-R1279.

    PubMed  Google Scholar 

  13. Bernoud N., Fenart, L., Molière P., Dehouck, M.P., Lagarde, M., Cecchelli, R., and Lecerf, J. (1999) Preferential Transfer of 2-Docosahexaenoyl-1-lysophosphatidylcholine Through an in vitro Blood-Brain Barrier Over Unesterified Docosahexaenoic Acid, J. Neurochem. 72, 338–345.

    Article  PubMed  CAS  Google Scholar 

  14. Slotboom, A.J., De Hass, G.H., Burbach-Westerhuis, G.J., and Van Deenen, L.L.M. (1970) Hydrolysis of Phosphoglycerides by Purified Lipase Preparations. Preparation of Unsaturated 2-Monoacyl Choline Phosphoglycerides, Chem. Phys. Lipids 4, 30–36.

    Article  CAS  Google Scholar 

  15. Bligh, E.G., and Dyer, W.J. (1959) A Rapid Method of Total Lipid Extraction and Purification, Can. J. Biochem. Physiol. 37, 911–917.

    PubMed  CAS  Google Scholar 

  16. Thiès, F., Delachambre, M.C., Bentejac, M., Lagarde, M., and Lecerf, J. (1992) Unsaturated Fatty Acids Esterified in 2-Acyl-1-lysophosphatidylcholine Bound to Albumin Are More Efficiently Taken Up by the Young Rat Brain Than the Unesterified Form, J. Neurochem. 59, 1110–1116.

    Article  PubMed  Google Scholar 

  17. Croset, M., Brossard, N., Polette, A., and Lagarde, M. (1999) Characterization of Plasma Unsaturated Lysophosphatidylcholines in Human and Rat, Biochem. J., in press.

  18. Robinson, B.S., Baisted, D.J., and Vance, D.E. (1989) Comparison of Albumin-Mediated Release of Lysophosphatidylcholine and Lysophosphatidylethanolamine from Cultured Rat Hepatocytes, Biochem. J. 264, 125–131.

    PubMed  CAS  Google Scholar 

  19. Gunstone, F.D., and Seth, S. (1994) A Study of the Distribution of Eicosapentaenoic Acid and Docosahexaenoic Acid Between the α and β Glycerol Chains in Fish Oils by 13C-NMR Spectroscopy, Chem. Phys. Lipids 71, 119–126.

    Article  Google Scholar 

  20. Sacchi, R., Medina, I., Aubourg, P.S., Giudicianni, I., Paolillo, L., and Addeo, F. (1993) Quantitative High-Resolution 13C NMR Analysis of Lipids Extracted from the White Muscle of Atlantic Tuna (Thunnus alalunga), J. Agric. Food Chem. 41, 1247–1253.

    Article  CAS  Google Scholar 

  21. Haraldsson, G.G., Gudmundsson, B.Ö., and Almarsson, Ö. (1995) The Synthesis of Homogeneous Triglycerides of Eicosapentaenoic Acid and Docosahexaenoic Acid by Lipase, Tetrahedron 51, 941–952.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. INSERM U352, Biochimie & Pharmacologie INSA-Lyon, 20 Ave. A. Einstein, 69621, Villeurbanne, France

    A. Polette, M. Croset & M. Lagarde

  2. Thallia-Pharmaceuticals Ecully, 20 Ave. A. Einstein, 69621, Villeurbanne, France

    A. Polette & J. M. Armstrong

  3. Chimie Organique INSA-Lyon, 20 Ave. A. Einstein, 69621, Villeurbanne, France

    C. Deshayes & B. Chantegrel

Authors
  1. A. Polette
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  2. C. Deshayes
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  3. B. Chantegrel
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  4. M. Croset
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  5. J. M. Armstrong
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  6. M. Lagarde
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Corresponding author

Correspondence to M. Lagarde.

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Polette, A., Deshayes, C., Chantegrel, B. et al. Synthesis of acetyl, docosahexaenoyl-glycerophosphocholine and its characterization using nuclear magnetic resonance. Lipids 34, 1333–1337 (1999). https://doi.org/10.1007/s11745-999-0486-1

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  • DOI: https://doi.org/10.1007/s11745-999-0486-1

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