, Volume 52, Issue 9, pp 751–761 | Cite as

Synthesis and Identification of AceDoxyPC, a Protectin-Containing Structured Phospholipid, Using Liquid Chromatography/Mass Spectrometry

  • Amanda Lo Van
  • Baptiste Fourmaux
  • Madeleine Picq
  • Michel Guichardant
  • Michel Lagarde
  • Nathalie Bernoud-HubacEmail author
Original Article


Fatty acids have many health benefits in a great variety of diseases ranging from cardiovascular to cerebral diseases. For instance, docosahexaenoic acid (DHA), which is highly enriched in brain phospholipids, plays a major role in anti-inflammatory or neuroprotective pathways. Its effects are thought to be due, in part, to its conversion into derived mediators such as protectins. 1-Lyso,2-docosahexaenoyl-glycerophosphocholine (LysoPtdCho-DHA) is one of the physiological carrier of DHA to the brain. We previously synthesized a structured phosphatidylcholine to mimic 1-lyso,2-docosahexaenoyl-glycerophosphocholine, named AceDoPC® (1-acetyl,2-docosahexaenoyl-glycerophosphocholine), that is considered as a stabilized form of the physiological LysoPtdCho-DHA and that is neuroprotective in experimental ischemic stroke. Considering these, the current study aimed at enzymatically oxygenate DHA contained within AceDoPC® to synthesize a readily structured oxidized phospholipid containing protectin DX (PDX), thereafter named AceDoxyPC (1-acetyl,2-PDX-glycerophosphocholine). Identification of this product was performed using liquid chromatography/tandem mass spectrometry. Such molecule could be used as a bioactive mediator for therapy against neurodegenerative diseases and stroke.


Omega-3 fatty acids Docosahexaenoic acid Lipoxygenase Mass spectrometry Protectin DX Structured phospholipid 







Blood–brain barrier


Collision-induced dissociation














Multiple reaction monitoring


Protectin D1


Protectin DX


Soybean lipoxygenase



This study was supported by Lisa CARNOT Institute, National Institute of Applied Sciences-Lyon and the French Ministry of Education and Research. We thank Laurence Daniel and Valentin Ramel for their technical and supportive assistance.

Compliance with Ethical Standards

Conflict of interest

Pending patent: WO 2017006047 A1. NBH, MP, MG, and ML are associates in LipTher (, an emerging start up from the academic CarMeN laboratory, INSA-Lyon, whose objective is to produce AceDoPC® on a large scale.


  1. 1.
    Thies F, Pillon C, Moliere P, Lagarde M, Lecerf J (1994) Preferential incorporation of sn-2 lysoPC DHA over unesterified DHA in the young rat brain. Am J Physiol 267:R1273–R1279PubMedGoogle Scholar
  2. 2.
    Bernoud N, Fenart L, Molière P, Dehouck MP, Lagarde M, Cecchelli R, Lecerf J (1999) Preferential transfer of 2-docosahexaenoyl-1-lysophosphatidylcholine through an in vitro blood–brain barrier over unesterified docosahexaenoic acid. J Neurochem 72:338–345CrossRefGoogle Scholar
  3. 3.
    Chen CT, Kitson AP, Hopperton KE, Domenichiello AF, Trépanier MO, Lin LE, Ermini L, Post M, Thies F, Bazinet RP (2015) Plasma non-esterified docosahexaenoic acid is the major pool supplying the brain. Sci Rep 5:15791CrossRefGoogle Scholar
  4. 4.
    Brossard N, Croset M, Lecerf J, Pachiaudi C, Normand S, Chirouze V, Macovschi O, Riou JP, Tayot JL, Lagarde M (1996) Metabolic fate of an oral tracer dose of [13C]docosahexaenoic acid triglycerides in the rat. Am J Physiol 270:R846–R854PubMedGoogle Scholar
  5. 5.
    Brossard N, Croset M, Normand S, Pousin J, Lecerf J, Laville M, Tayot JL, Lagarde M (1997) Human plasma albumin transports [13C]docosahexaenoic acid in two lipid forms to blood cells. J Lipid Res 7:1571–1582Google Scholar
  6. 6.
    Nguyen NL, Ma D, Shui G, Wong P, Cazenave-Gassiot A, Zhang X, Wenk MR, Goh ELK, Silver DL (2014) Mfsd2a is a transporter for the essential omega-3 fatty acid docosahexaenoic acid. Nature 509:503–506CrossRefGoogle Scholar
  7. 7.
    Quek DQY, Nguyen LN, Fan H, Silver DL (2016) Structural insights into the transport mechanism of the human sodium-dependent lysophosphatidylcholine transporter MFSD2A. J Biol Chem 291:9383–9394CrossRefGoogle Scholar
  8. 8.
    Wong BH, Chan JP, Cazenave-Gassiot A, Poh RW, Foo JC, Galam DL, Ghosh S, Nguyen LN, Barathi VA, Yeo SW, Luu CD, Wenk MR, Silver DL (2016) Mfsd2a is a transporter for the essential ω-3 fatty acid docosahexaenoic acid (DHA) in eye and is important for photoreceptor cell development. J Biol Chem 291:10501–10514CrossRefGoogle Scholar
  9. 9.
    Angers M, Uldry M, Kong D, Gimble JM, Jetten AM (2008) Mfsd2a encodes a novel major facilitator superfamily domain-containing protein highly induced in brown adipose tissue during fasting and adaptive thermogenesis. Biochem J 416:347–355CrossRefGoogle Scholar
  10. 10.
    Croset M, Brossard N, Polette A, Lagarde M (2000) Characterization of plasma unsaturated lysophosphatidylcholines in human and rat. Biochem J 345:61–67CrossRefGoogle Scholar
  11. 11.
    Polette A, Deshayes C, Chantegrel B, Croset M, Armstrong JM, Lagarde M (1999) Synthesis of acetyl, docosahexaenoyl-glycerophosphocholine and its characterization using nuclear magnetic resonance. Lipids 34:1333–1337CrossRefGoogle Scholar
  12. 12.
    Lagarde M, Hachem M, Bernoud-Hubac N, Picq M, Véricel E, Guichardant M (2015) Biological properties of a DHA-containing structured phospholipid (AceDoPC) to target the brain. Prostaglandins Leukot Essent Fat Acids 92:63–65CrossRefGoogle Scholar
  13. 13.
    Hachem M, Géloën A, Lo Van A, Fourmaux B, Fenart L, Gosselet F, Da Silva P, Breton G, Lagarde M, Picq M, Bernoud-Hubac N (2016) Efficient docosahexaenoic acid uptake by the brain from a structured phospholipid. Mol Neurobiol 53:3205–3215CrossRefGoogle Scholar
  14. 14.
    Chauveau F, Cho T-H, Perez M, Guichardant M, Riou A, Aguettaz P, Picq M, Lagarde M, Berthezène Y, Nighoghossian N, Wiart M (2011) Brain-targeting form of docosahexaenoic acid for experimental stroke treatment: MRI evaluation and anti-oxidant impact. Curr Neurovasc Res 8:95–102CrossRefGoogle Scholar
  15. 15.
    Lo Van A, Sakayori N, Hachem M, Belkouch M, Picq M, Lagarde M, Osumi N, Bernoud-Hubac N (2016) Mechanisms of DHA transport to the brain and potential therapy to neurodegenerative diseases. Biochimie 130:163–167CrossRefGoogle Scholar
  16. 16.
    Belkouch M, Hachem M, Elgot A, Lo A, Picq M, Guichardant M, Lagarde M, Bernoud-Hubac N (2016) The pleiotropic effects of omega-3 docosahexaenoic acid on the hallmarks of Alzheimer’s disease. J Nutr Biochem 38:1–11CrossRefGoogle Scholar
  17. 17.
    Serhan CN (2005) Mediator lipidomics. Prostaglandins Other Lipid Mediat 77:4–14CrossRefGoogle Scholar
  18. 18.
    Gonzalez-Periz A, Horrillo R, Ferre N, Gronert K, Dong B, Moran-Salvador E, Titos E, Martinez-Clemente M, Lopez-Parra M, Arroyo V, Claria J (2009) Obesity-induced insulin resistance and hepatic steatosis are alleviated by omega-3 fatty acids: a role for resolvins and protectins. FASEB J 23:1946–1957CrossRefGoogle Scholar
  19. 19.
    Aveldano MI, Sprecher H (1983) Synthesis of hydroxy fatty acids from 4,7,10,13,16,19-[1-14C] docosahexaenoic acid by human platelets. J Biol Chem 25:9339–9343Google Scholar
  20. 20.
    Miller C, Yamaguchi RY, Ziboh VA (1989) Guinea pig epidermis generates putative anti-inflammatory metabolites from fish oil polyunsaturated fatty acids. Lipids 24:998–1003CrossRefGoogle Scholar
  21. 21.
    Kim HY, Karanian JW, Salem NJ (1990) Formation of 15-lipoxygenase product from docosahexaenoic acid (22:6w3) by human platelets. Prostaglandins 40:539–549CrossRefGoogle Scholar
  22. 22.
    Chen P, Fenet B, Michaud S, Tomczyk N, Véricel E, Lagarde M, Guichardant M (2009) Full characterization of PDX, a neuroprotectin/protectin D1 isomer, which inhibits blood platelet aggregation. FEBS Lett 583:3478–3484CrossRefGoogle Scholar
  23. 23.
    Balas L, Guichardant M, Durand T, Lagarde M (2014) Confusion between protectin D1 (PD1) and its isomer protectin DX (PDX). An overview on the dihydroxy-docosatrienes described to date. Biochimie 99:1–7CrossRefGoogle Scholar
  24. 24.
    Chen P, Véricel E, Lagarde M, Guichardant M (2011) Poxytrins, a class of oxygenated products from polyunsaturated fatty acids, potently inhibit blood platelet aggregation. FASEB J 25:382–388CrossRefGoogle Scholar
  25. 25.
    Huang LS, Hung ND, Sok DE, Kim MR (2010) Lysophosphatidylcholine containing docosahexaenoic acid at the sn-1 position is anti-inflammatory. Lipids 45:225–236CrossRefGoogle Scholar
  26. 26.
    Bochkov VN, Kadl A, Huber J, Gruber F, Binder BR, Leitinger N (2002) Protective role of phospholipid oxidation products in endotoxin-induced tissue damage. Lett Nat 4:77–81CrossRefGoogle Scholar
  27. 27.
    Leitinger N, Tyner TR, Oslund L, Rizza C, Subbanagounder G, Lee H, Shih PT, Mackman N, Tigyi G, Territo MC, Berliner JA, Vora DK (1999) Structurally similar oxidized phospholipids differentially regulate endothelial binding of monocytes and neutrophils. Proc Natl Acad Sci USA 96:12010–12015CrossRefGoogle Scholar
  28. 28.
    Murray JJ, Brash AR (1988) Rabbit reticulocyte lipoxygenase catalyzes specific 12(S) and 15(S) oxygenation of arachidonoyl-phosphatidylcholine. Arch Biochem Biophys 265:514–523CrossRefGoogle Scholar
  29. 29.
    Takahashi Y, Glasgow WC, Suzuki H, Taketani Y, Yamamoto S, Anton M, Kühn H, Brash AR (1993) Investigation of the oxygenation of phospholipids by the porcine leukocyte and human platelet arachidonate 12-lipoxygenases. Eur J Biochem 218:165–171CrossRefGoogle Scholar
  30. 30.
    Maccarrone M, van Aarle PGM, Veldink GA, Vliegenthart JFG (1994) In vitro oxygenation of soybean biomembranes by lipoxygenase-2. BBA Biomembr 1190:164–169CrossRefGoogle Scholar
  31. 31.
    Huang LS, Kim MR, Sok D-E (2007) Oxygenation of 1-docosahexaenoyl lysophosphatidylcholine by lipoxygenases; conjugated hydroperoxydiene and dihydroxytriene derivatives. Lipids 42:981–990CrossRefGoogle Scholar
  32. 32.
    Bochkov VN, Oskolkova OV, Birukov KG, Levonen AL, Binder CJ, Stöckl J (2010) Generation and biological activities of oxidized phospholipids. Antioxid Redox Signal 12:1009–1059CrossRefGoogle Scholar
  33. 33.
    Ashraf MZ, Srivastava S (2012) Oxidized phospholipids: introduction and biological significance. In: Frank S, Kostner G (eds) Lipoproteins – role health disease. InTech, Rijeka, pp 409–430Google Scholar
  34. 34.
    Butovich IA, Lukyanova SM, Bachmann C (2006) Dihydroxydocosahexaenoic acids of the neuroprotectin D family: synthesis, structure, and inhibition of human 5-lipoxygenase. J Lipid Res 47:2462–2474CrossRefGoogle Scholar

Copyright information

© AOCS 2017

Authors and Affiliations

  • Amanda Lo Van
    • 1
    • 2
  • Baptiste Fourmaux
    • 1
  • Madeleine Picq
    • 1
  • Michel Guichardant
    • 1
  • Michel Lagarde
    • 1
  • Nathalie Bernoud-Hubac
    • 1
    Email author
  1. 1.Univ Lyon, INSA-Lyon, Inserm UMR 1060, Inra UMR 1397, CarMeN Laboratory, INSA, Bâtiment IMBLVilleurbanne CedexFrance
  2. 2.Department of Developmental Neuroscience, Center for Neuroscience, ARTTohoku University Graduate School of MedicineSendaiJapan

Personalised recommendations