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High Pressure Liquid Chromatography of Eicosanoids

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Biochemistry of Arachidonic Acid Metabolism

Part of the book series: Prostaglandins, Leukotrienes, and Cancer ((PLAC,volume 1))

Abstract

Arachidonic acid (20:4) and other polyunsaturated fatty acids can be oxygenated in many tissues by cyclooxygenase or a variety of lipoxygenases. The intermediates formed in these reactions are then converted enzymatically to a variety of other products. In many cases large numbers of closely-related substances are formed in these reactions, necessitating the use of sophisticated methods for their separation and analysis. High pressure liquid chromatography (HPLC) is ideally suited for this purpose. The use of this technique has been very important for the identification and analysis of a large number of recently discovered 20:4 metabolites, including leukotrienes (1,2) and epoxy and dihydroxy products formed by the actions of epoxygenases related to cytochrome P-450 (3,4). The utility of HPLC is due to the improved chromatography achieved by the use of stationary phases composed of very small and uniform particles. This minimizes peak broadening due to differences in the paths followed by solute molecules as they pass through the column (Eddy diffusion). Consequently, the efficiency of a column increases as the particle size decreases.

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References

  1. Borgeat P, Samuelsson B: Arachidonic acid metabolism in polymorphonuclear leukocytes: effects of ionophore A23187. Proc Natl Acad Sci USA (76): 2148–2152, 1979.

    Article  PubMed  CAS  Google Scholar 

  2. Murphy RC, Hammmarström S, Samuelsson B: Leukotriene C: a slow-reacting substance from murine mastocytoma cells. Proc Natl Acad Sci USA (76): 4275–4279, 1979.

    Article  PubMed  CAS  Google Scholar 

  3. Oliw EH, Guengerich FP, Oates JA: Oxygenation of arachidonic acid by hepatic monooxygenases. Isolation and metabolism of four epoxide intermediates. J Biol Chem (257): 3771–3782, 1982.

    PubMed  CAS  Google Scholar 

  4. Chacos N, Falck JR, Wixtrom C, Capdevila J: Novel epoxides formed during the liver cytochrome P-450 oxidation of arachidonic acid, Biochem Biophys Res Commun (104): 916–922, 1982.

    Article  PubMed  CAS  Google Scholar 

  5. Powell WS: Rapid extraction of oxygenated metabolites of arachidonic acid from biological samples using octadecylsilyl silica. Prostaglandins (20): 947–957, 1980.

    Article  PubMed  CAS  Google Scholar 

  6. Powell WS: Properties of leukotriene B4 20-hydroxylase from polymorphonuclear leukocytes. J Biol Chem (259): 3082–3089, 1984.

    PubMed  CAS  Google Scholar 

  7. Hamberg M, Samuelsson B: On the specificity of the oxygenation of unsaturated fatty acids catalyzed by soybean lipoxidase. J Biol Chem (242): 5329–5335, 1967.

    PubMed  CAS  Google Scholar 

  8. Boeynaems JM, Brash AR, Oates JA, Hubbard WC: Preparation and assay of monohydroxyeicosatetraenoic acids. Anal Biochem (104): 259–267, 1980.

    Article  PubMed  CAS  Google Scholar 

  9. Powell WS: Formation of 6-oxoprostaglandin F1α , 6,15-dioxoprostaglandin F1α , and monohydroxyicosatetraenoic acids from arachidonic acid by fetal calf aorta and ductus arteriosus. J Biol Chem (257): 9457–9464, 1982.

    PubMed  CAS  Google Scholar 

  10. Powell WS: Formation of trihydroxyheptadecenoic acids, monohydroxyicosatrienoic acids, and prostaglandins from 8,11,14-icosatrienoic acid by adult and fetal aorta. J Biol Chem (257): 9465–9472, 1982.

    PubMed  CAS  Google Scholar 

  11. Borgeat P, Samuelsson B: Transformation of arachidonic acid by rabbit polymorphonuclear leukocytes. Formation of a novel dihydroxyeicosatetraenoic acid. J Biol Chem (254): 2643–2646, 1979.

    PubMed  CAS  Google Scholar 

  12. Borgeat P, Samuelsson B: Metabolism of arachidonic acid in polymorphonuclear leukocytes. Structural analysis of novel hydroxylated compounds. J Biol Chem (254) 7865–7869, 1979.

    PubMed  CAS  Google Scholar 

  13. Borgeat P, Picard S, Vallerand P, Sirois P: Transformation of arachidonic acid in leukocytes. Isolation and structural analysis of a novel dihydroxy derivative. Prostaglandins Med (6): 557–570, 1981.

    Article  PubMed  CAS  Google Scholar 

  14. Maas RL, Turk J, Oates JA, Brash AR: Formation of a novel dihydroxy acid from arachidonic acid by lipoxygenase-catalyzed double oxygenation in rat mononuclear cells and human Leukocytes. J Biol Chem (257): 7056–7067, 1982.

    PubMed  CAS  Google Scholar 

  15. Orning L, Hammarström S, Samuelsson B: Leukotriene D: a slow reacting substance from rat basophilic leukemia cells. Proc Natl Acad Sci USA (77): 2014–2017, 1980.

    Article  PubMed  CAS  Google Scholar 

  16. Lewis RA, Drazen JM, Austen KF, Clark DA, Corey EJ: Identification of the C(6)-S-conjugate of leukotriene A with cysteine as a naturally occurring slow reacting substance of anaphylaxis (SRS-A). Importance of the 11-cis geometry for biological activity. Biochem Biophys Res Commun (96): 271–277, 1980.

    Article  PubMed  CAS  Google Scholar 

  17. Borgeat P, Samuelsson B: Transformation of arachidonic acid and homo-γ-linolenic acid by rabbit polymorphonuclear leukocytes. J Biol Chem (251): 7816–7820, 1976.

    PubMed  CAS  Google Scholar 

  18. Hamberg M, Samuelsson B: Prostaglandin endoperoxides. Novel tranformations of arachidonic acid in human platelets. Proc Natl Acad Sci USA (71) 3400–3404, 1974.

    Article  PubMed  CAS  Google Scholar 

  19. Bergström S, Ryhage R, Samuelsson B, Sjovall J: Prostaglandins and related factors 15. The structure of prostaglandins E1, F1α , and F1β . J Biol Chem (238): 3555–3564, 1963.

    Google Scholar 

  20. Morozowich W, Douglas SL: Resolution of prostaglandin p-nitrophenacyl esters by liquid chromatography and conditions for rapid, quantitative p-nitrophenacylation. Prostaglandins (10): 19–40, 1975.

    PubMed  CAS  Google Scholar 

  21. Fitzpatrick F: High performance liquid chromatographic determination of prostaglandins F2α , E2 and D2 from in vitro enzyme incubations. Anal Chem (48): 499–502, 1976.

    Article  PubMed  CAS  Google Scholar 

  22. Turk J, Weiss SJ, Davis JE, Needleman P: Fluorescent derivatives of prostaglandins and thromboxanes for liquid chromatography. Prostaglandins (16): 291–309, 1978.

    Article  PubMed  CAS  Google Scholar 

  23. Hatsumi M, Kimata SI, Hirosawa K: 9-anthryldiazomethane derivatives of prostaglandins for high-performance liquid chromatographic analysis. J Chromatogr (253) 271–275, 1982.

    Article  CAS  Google Scholar 

  24. Gréen K, Hamberg M, Samuelsson B, Frölich J: Extraction and chromatographic procedures for purification of prostaglandins, thromboxanes, prostacyclin, and their metabolites. Adv Prostaglandin Thromboxane Res (5): 15–38, 1978.

    PubMed  Google Scholar 

  25. Powell WS: Rapid extraction of arachidonic acid metabolites from biological samples using octadecylsilyl silica. Methods Enzymol (86): 467–477, 1982.

    Article  PubMed  CAS  Google Scholar 

  26. Bennett HPJ, Hudson AM, McMartin C, Purdon GE: Use of octadecylsilyl silica for the extraction and purification of peptides in biological samples, Biochem J (168): 9–13, 1977.

    PubMed  CAS  Google Scholar 

  27. Mathews WR, Rokach J, Murphy RC: Analysis of leukotrienes by high-pressure liquid chromatography. Anal Biochem (118): 96–101, 1981.

    Article  PubMed  CAS  Google Scholar 

  28. Metz SA, Hall ME, Harper TW, Murphy RC: Rapid extraction of leukotrienes from biological fluids and quantitation by high-performance liquid chromatography. J Chromatogr (233): 193–201, 1982.

    Article  PubMed  CAS  Google Scholar 

  29. Borgeat P, Fruteau de Laclos B, Rabinovitch H, Picard S, Braquet P, Hebert J, Laviolette M: Eosinophil-rich human polymorphonuclear leukocyte preparations characteristically release leukotriene C4 upon ionophore A23187 challenge. J Allergy Clin Immunol, in press.

    Google Scholar 

  30. Borgeat P: Reversed-phase high-pressure liquid chromatography profiling and quantitation of lipoxygenase products. Prostaglandins, in press.

    Google Scholar 

  31. Porter NA, Logan J, Kontoyiannidou V: Preparaation and purification of arachidonic acid hydroperoxides of biological importance. J Org Chem (44): 3177–3181, 1979.

    Article  CAS  Google Scholar 

  32. Maclouf J, Fruteau de Laclos B, Borgeat P: Stimulation of leukotriene biosynthesis in human blood leukocytes by platelet-derived 12-hydroperoxyicosatetraenoic acid. Proc Natl Acad Sci USA (79): 6042–6046, 1982.

    Article  PubMed  CAS  Google Scholar 

  33. Alam I, Ohuchi I, Levine L: Determination of cyclooxygenase products and prostaglandin metabolites using high-pressure liquid chromatography and radioimmunoassay. Anal Biochem (93): 339–345, 1979.

    Article  PubMed  CAS  Google Scholar 

  34. Eling T, Tainer B, Ally A, Warnock R: Separation of arachidonic acid metabolites by high-pressure liquid chromatography. Methods Enzymol (86): 511–517, 1982.

    Article  PubMed  CAS  Google Scholar 

  35. Van Rollins M, Aveldano MI, Sprecher HW, Horrocks LA: High-pressure liquid chromatography of underivatized fatty acids, hydroxy acids, and prostanoids having different chain lengths and double-bond positions. Methods Enzymol (86): 518–530, 1982.

    Article  PubMed  Google Scholar 

  36. Metz SA, Hall ME, Harper TW, Murphy RC: Rapid extraction of leukotrienes from biological fluids and quantitation by high-performance liquid chromatography. J Chromatogr (275): 468, 1983.

    Article  PubMed  CAS  Google Scholar 

  37. Powell WS: Separation of icosenoic acids, monohydroxyicosenoic acids, and prostaglandins by high-pressure liquid chromatography on a silver ion-loaded cation exchange column. Anal Biochem (115): 267–277, 1981.

    Article  PubMed  CAS  Google Scholar 

  38. Powell WS: Argentation-high-pressure liquid chromatography of prostaglandins and monohydroxyeicosatetraenoic acids. Methods Enzymol (86): 530–543, 1982.

    Article  CAS  Google Scholar 

  39. Powell WS: Separation of unlabeled and isotopically-labeled metabolites of arachidonic acid by argentation high-pressure liquid chromatography. Adv Prostaglanin Thromboxane Leukotriene Res (11): 207–213, 1983.

    CAS  Google Scholar 

  40. Powell WS: Separation of unlabeled metabolites of arachidonic acid from their deuterium- and tritium-labeled analogs by argentation high-pressure liquid chromatography. Anal Biochem (128): 93–103, 1983.

    Article  PubMed  CAS  Google Scholar 

  41. Sun FF, McGuire, JC: Inhibition of human neutrophil arachidonate 5-lipoxygenase by 6,9-deepoxy-6,9-(phenylimino)-Δ6,8-prostaglandin I1, (U-60257). Prostaglandins (26): 211–221, 1983.

    Article  PubMed  CAS  Google Scholar 

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Authors
  1. William S. Powell
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Editors and Affiliations

  1. Department of Biological Chemistry College of Medicine, University of Illinois at Chicago, 1853 W. Polk St., 60612, Chicago, IL, USA

    William E. M. Lands

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© 1985 Martinus Nijhoff Publishing, Boston

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Powell, W.S. (1985). High Pressure Liquid Chromatography of Eicosanoids. In: Lands, W.E.M. (eds) Biochemistry of Arachidonic Acid Metabolism. Prostaglandins, Leukotrienes, and Cancer, vol 1. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-2597-0_22

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  • DOI: https://doi.org/10.1007/978-1-4613-2597-0_22

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4612-9627-0

  • Online ISBN: 978-1-4613-2597-0

  • eBook Packages: Springer Book Archive

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