Clinical and Experimental Medicine

, Volume 15, Issue 3, pp 397–403

LC–MS/MS analysis of plasma polyunsaturated fatty acids in patients with homozygous sickle cell disease

  • Mutay Aslan
  • Gamze Celmeli
  • Filiz Özcan
  • Alphan Kupesiz
Original Article


The aim of this study was to determine circulating omega-6, omega-3 polyunsaturated fatty acids and prostaglandin E2 (PGE2) levels in steady state sickle cell disease (SCD) patients. Blood was collected from healthy hemoglobin volunteers and steady state homozygous HbSS patients who had not received blood transfusions in the last 3 months. Plasma levels of arachidonic acid (AA, C20:4n-6), dihomo-gamma-linolenic acid (DGLA, C20:3n-6), eicosapentaenoic acid (EPA, C20:5n-3) and docosahexaenoic acid (DHA, C22:6n-3) were determined by an optimized multiple reaction monitoring method using ultrafast liquid chromatography coupled with tandem mass spectrometry. PGE2 was measured in serum samples by enzyme immunoassay. Plasma AA and DGLA were significantly increased while EPA and DHA were significantly decreased in SCD plasma compared to control. Serum PGE2 levels, AA/DHA and AA/EPA ratio was significantly higher in SCD patients when compared to control group. The significant increase in PGE2 levels, AA/EPA and AA/DHA ratio confirms the presence of a proinflammatory state in SCD patients.


Sickle cell disease Polyunsaturated fatty acid Prostaglandin E2 


  1. 1.
    Aslan M, Freeman BA (2007) Redox-dependent impairment of vascular function in sickle cell disease. Free Radic Biol Med 43(11):1469–1483CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Hebbel RP (1997) Adhesive interactions of sickle erythrocytes with endothelium. J Clin Invest 100(11 Suppl):S83–S86PubMedGoogle Scholar
  3. 3.
    Brown MD, Wick TM, Eckman JR (2001) Activation of vascular endothelial cell adhesion molecule expression by sickle blood cells. Pediatr Pathol Mol Med 20(1):47–72CrossRefPubMedGoogle Scholar
  4. 4.
    Platt OS, Brambilla DJ, Rosse WF, Milner PF, Castro O, Steinberg MH, Klug PP (1994) Mortality in sickle cell disease. Life expectancy and risk factors for early death. N Engl J Med 330(23):1639–1644CrossRefPubMedGoogle Scholar
  5. 5.
    Castro O, Brambilla DJ, Thorington B, Reindorf CA, Scott RB, Gillette P, Vera JC, Levy PS (1994) The acute chest syndrome in sickle cell disease: incidence and risk factors. The cooperative study of sickle cell disease. Blood 84(2):643–649PubMedGoogle Scholar
  6. 6.
    Kinney TR, Sleeper LA, Wang WC, Zimmerman RA, Pegelow CH, Ohene-Frempong K, Wethers DL, Bello JA, Vichinsky EP, Moser FG, Gallagher DM, DeBaun MR, Platt OS, Miller ST (1999) Silent cerebral infarcts in sickle cell anemia: a risk factor analysis. The cooperative study of sickle cell disease. Pediatrics 103(3):640–645CrossRefPubMedGoogle Scholar
  7. 7.
    Bates EJ, Ferrante A, Smithers L, Poulos A, Robinson BS (1995) Effect of fatty acid structure on neutrophil adhesion, degranulation and damage to endothelial cells. Atherosclerosis 116(2):247–259CrossRefPubMedGoogle Scholar
  8. 8.
    De Caterina R, Liao JK, Libby P (2000) Fatty acid modulation of endothelial activation. Am J Clin Nutr 71(1 Suppl):213S–223SPubMedGoogle Scholar
  9. 9.
    Wall R, Ross RP, Fitzgerald GF, Stanton C (2010) Fatty acids from fish: the anti-inflammatory potential of long-chain omega-3 fatty acids. Nutr Rev 68(5):280–289CrossRefPubMedGoogle Scholar
  10. 10.
    Serhan CN, Chiang N, Van Dyke TE (2008) Resolving inflammation: dual anti-inflammatory and pro-resolution lipid mediators. Nat Rev Immunol 8(5):349–361CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Connor WE, Lin DS, Thomas G, Ey F, DeLoughery T, Zhu N (1997) Abnormal phospholipid molecular species of erythrocytes in sickle cell anemia. J Lipid Res 38(12):2516–2528PubMedGoogle Scholar
  12. 12.
    Ren H, Okpala I, Ghebremeskel K, Ugochukwu CC, Ibegbulam O, Crawford M (2005) Blood mononuclear cells and platelets have abnormal fatty acid composition in homozygous sickle cell disease. Ann Hematol 84(9):578–583CrossRefPubMedGoogle Scholar
  13. 13.
    Aslan M, Ozcan F, Aslan B, Yücel G (2013) LC-MS/MS analysis of plasma polyunsaturated fatty acids in type 2 diabetic patients after insulin analog initiation therapy. Lipids Health Dis 12(1):169CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Aslan M, Aslan I, Ozcan F, Eryılmaz R, Ensari CO, Bilecik T (2014) A pilot study investigating early postoperative changes of plasma polyunsaturated fatty acids after laparoscopic sleeve gastrectomy. Lipids Health Dis 13(1):62CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Ou CN, Rognerud CL (2001) Diagnosis of hemoglobinopathies: electrophoresis vs. HPLC. Clin Chim Acta 313(1–2):187–194CrossRefPubMedGoogle Scholar
  16. 16.
    Rees DC, Williams TN, Gladwin MT (2010) Sickle-cell disease. Lancet 376(9757):2018–2031CrossRefPubMedGoogle Scholar
  17. 17.
    Akinsheye I, Alsultan A, Solovieff N, Ngo D, Baldwin CT, Sebastiani P, Chui DH, Steinberg MH (2011) Fetal hemoglobin in sickle cell anemia. Blood 118(1):19–27CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Nagel RL, Bookchin RM, Johnson J, Labie D, Wajcman H, Isaac-Sodeye WA, Honig GR, Schilirò G, Crookston JH, Matsutomo K (1979) Structural bases of the inhibitory effects of hemoglobin F and hemoglobin A2 on the polymerization of hemoglobin S. Proc Natl Acad Sci U S A 76(2):670–672CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Shokrani M, Terrell F, Turner EA, Aguinaga MD (2000) Chromatographic measurements of hemoglobin A2 in blood samples that contain sickle hemoglobin. Ann Clin Lab Sci 30(2):191–194PubMedGoogle Scholar
  20. 20.
    Mohan JS, Lip GY, Bareford D, Blann AD (2006) Platelet P-selectin and platelet mass, volume and component in sickle cell disease: relationship to genotype. Thromb Res 117(6):623–629CrossRefPubMedGoogle Scholar
  21. 21.
    Tomer A, Harker LA, Kasey S, Eckman JR (2001) Thrombogenesis in sickle cell disease. J Lab Clin Med 137(6):398–407CrossRefPubMedGoogle Scholar
  22. 22.
    Calder PC (2001) Polyunsaturated fatty acids, inflammation, and immunity. Lipids 36(9):1007–1024CrossRefPubMedGoogle Scholar
  23. 23.
    Setty BN, Dampier CD, Stuart MJ (1995) Arachidonic acid metabolites are involved in mediating red blood cell adherence to endothelium. J Lab Clin Med 125(5):608–617PubMedGoogle Scholar
  24. 24.
    Stuart MJ, Sills RH (1981) Deficiency of plasma prostacyclin or PGI2 regenerating ability in sickle cell anaemia. Br J Haematol 48(4):545–550CrossRefPubMedGoogle Scholar
  25. 25.
    Setty BN, Chen D, Stuart MJ (1995) Sickle cell vaso-occlusive crisis is associated with abnormalities in the ratio of vasoconstrictor to vasodilator prostanoids. Pediatr Res 38(1):95–102CrossRefPubMedGoogle Scholar
  26. 26.
    Daak AA, Ghebremeskel K, Hassan Z, Attallah B, Azan HH, Elbashir MI, Crawford M (2013) Effect of omega-3 (n-3) fatty acid supplementation in patients with sickle cell anemia: randomized, double-blind, placebo-controlled trial. Am J Clin Nutr 97(1):37–44CrossRefPubMedGoogle Scholar
  27. 27.
    Daak AA, Ghebremeskel K, Mariniello K, Attallah B, Clough P, Elbashir MI (2013) Docosahexaenoic and eicosapentaenoic acid supplementation does not exacerbate oxidative stress or intravascular haemolysis in homozygous sickle cell patients. Prostaglandins Leukot Essent Fatty Acids 89(5):305–311CrossRefPubMedGoogle Scholar
  28. 28.
    Aslan M, Canatan D (2008) Modulation of redox pathways in neutrophils from sickle cell disease patients. Exp Hematol 36(11):1535–1544CrossRefPubMedGoogle Scholar
  29. 29.
    Kupesiz A, Celmeli G, Dogan S, Antmen B, Aslan M (2012) The effect of hemolysis on plasma oxidation and nitration in patients with sickle cell disease. Free Radic Res 46(7):883–890CrossRefPubMedGoogle Scholar
  30. 30.
    Aslan M, Freeman BA (2004) Oxidant-mediated impairment of nitric oxide signaling in sickle cell disease–mechanisms and consequences. Cell Mol Biol (Noisy-le-grand) 50(1):95–105Google Scholar
  31. 31.
    Graido-Gonzalez E, Doherty JC, Bergreen EW, Organ G, Telfer M, McMillen MA (1998) Plasma endothelin-1, cytokine, and prostaglandin E2 levels in sickle cell disease and acute vaso-occlusive sickle crisis. Blood 92(7):2551–2555PubMedGoogle Scholar

Copyright information

© Springer-Verlag Italia 2014

Authors and Affiliations

  • Mutay Aslan
    • 1
  • Gamze Celmeli
    • 2
  • Filiz Özcan
    • 1
  • Alphan Kupesiz
    • 2
  1. 1.Department of Medical BiochemistryAkdeniz University Medical SchoolAntalyaTurkey
  2. 2.Department of Pediatric HematologyAkdeniz University Medical SchoolAntalyaTurkey

Personalised recommendations