European Journal of Nutrition

, Volume 54, Issue 8, pp 1287–1295 | Cite as

Inhibitory and synergistic effects of natural olive phenols on human platelet aggregation and lipid peroxidation of microsomes from vitamin E-deficient rats

  • Fátima Rubio-Senent
  • Baukje de Roos
  • Garry Duthie
  • Juan Fernández-Bolaños
  • Guillermo Rodríguez-GutiérrezEmail author
Original Contribution



This study explored the in vitro antioxidant and anti-platelet activities of hydroxytyrosol, hydroxytyrosol acetate, 3,4-dihydroxyphenylglycol and two phenolic olive extracts. These compounds and extracts were obtained from a new industrial process to hydrothermally treat the alperujo (160 °C/60 min), a by-product of olive oil extraction.


The extracts and the purified compounds were obtained chromatographically using both ionic and adsorbent resins. The antioxidant activity was determined by measuring inhibition of human platelet aggregation and inhibition of lipid peroxidation in liver microsomes of vitamin E-deficient rats.


The positive effect of the extracts on the inhibition of platelet aggregation is showed, being higher in the case of hydroxytyrosol acetate up to 38 %, and for the first time, its synergist effect with hydroxytyrosol has been proved, obtaining more than double of inhibition. The phenolic extracts and the isolated phenols showed good results for inhibiting the lipid oxidation, up to 62 and 25 %, respectively. A synergistic effect occurred when the hydroxytyrosol acetate and the 3,4-dihydroxyphenylglycol were supplemented by hydroxytyrosol.


These results suggest the extract and these compounds obtained from a novel industrial process could be natural alternatives for the prevention of diseases related to cardiovascular disorder or oxidative damage.


Alperujo Olive oil wastes Phenols Platelet function Lipid peroxidation Vitamin E 



The authors wish to express their gratitude to the Ministerio de Economía y Competitividad of Spain and co-funded by European Social Fund (ESF) (Project AGL2013-48291-R) for providing financial support. Dr. Rodríguez-Gutiérrez (RYC-2012-10456 contract) wishes to thank to the “Ramón y Cajal” Program from the Spanish Ministry of Economy and Competitiveness, and Ms Rubio-Senent (JAE-Pre104) to the Spanish National Research Council and co-funded by European Social Fund (CSIC-ESF) for providing financial support.

Conflict of interest



  1. 1.
    Estruch R, Ros E, Salas-Salvado J, Covas MI, Corella D, Aros F, Gómez-Gracia E, Ruíz-Gutiérrez V et al (2013) Prevention of cardiovascular disease with a Mediterranean diet. New Engl J Med 368:1279–1290CrossRefGoogle Scholar
  2. 2.
    Ruiz-Canela M, Estruch R, Corella D, Salas-Salvado J, Martínez-González MA (2014) Association of Mediterranean diet with peripheral artery disease: the PREDIMED randomized trial. J Am Med Assoc 311:415–417CrossRefGoogle Scholar
  3. 3.
    González-Santiago M, Martín-Bautista E, Carrero JJ, Fonolla J, Baro L, Bartolomé MV, Gil-Loyzaga P, López-Huertas E (2006) One-month administration of hidroxitirosol, a phenolic anti-oxidant present in olive oil, to hyperlipemic rabbits improve blood lipid profile, anti-oxidant status and reduces atherosclerosis development. Atherosclerosis 188:35–42CrossRefGoogle Scholar
  4. 4.
    Visioli F, Poli A, Galli C (2002) Anti-oxidant and other biological activities of phenols from olives and olive oil. Med Res Rev 22:65–75CrossRefGoogle Scholar
  5. 5.
    Ostertag LM, O’Kennedy N, Kroon PA, Duthie GG, de Roos B (2010) Impact of dietary polyphenols on human platelet function—a critical review of controlled dietary intervention studies. Mol Nutr Food Res 54:60–81CrossRefGoogle Scholar
  6. 6.
    De Roos B, Zhang X, Rodríguez-Gutiérrez G, Wood S, Rucklidge GJ, Reid MD, Duncan GJ, Cantlay LL, Duthie GG, O’Kennedy N (2011) Anti-platelet effects of olive oil extract: in vitro functional and proteomic studies. Eur J Nutr 50:553–562CrossRefGoogle Scholar
  7. 7.
    Duthie GG and Morrice P (2012) Anti-oxidant capacity of flavonoids in hepatic microsomes is not reflected by anti-oxidant effects in vivo. Oxid Med Cell Longev 165127Google Scholar
  8. 8.
    Aruoma OI (1998) Free radicals, oxidative stress, and anti-oxidants in human health and disease. J Am Oil Chem Soc 75:199–212CrossRefGoogle Scholar
  9. 9.
    Fernández-Bolaños J, Rodríguez G, Gómez E, Guillén R, Jiménez A, Heredia A, Rodríguez R (2004) Total recovery of the waste of two-phase olive oil processing: isolation of added-value compounds. J Agric Food Chem 52:5849–5855CrossRefGoogle Scholar
  10. 10.
    Rubio-Senent F, Rodríguez-Gutiérrez G, Lama-Munoz A, Fernández-Bolaños J (2012) New phenolic compounds hydrothermally extracted from the olive oil by-product alperujo and their antioxidative activities. J Agric Food Chem 60:1175–1186CrossRefGoogle Scholar
  11. 11.
    Fernández-Bolaños J, Rodríguez G, Lama-Muñoz A, Sánchez P (2011) Dispositivo y procedimiento para el tratamiento de los subproductos de la obtención del aceite de oliva. Patent Request No. PCT/ES2011/070583Google Scholar
  12. 12.
    Rodríguez-Gutiérrez G, Duthie GG, Wood S, Morrice P, Nicol F, Reid M, Cantlay LL, Kelder T, Horgan GW, Fernández-Bolaños J, de Roos B (2012) Alperujo extract, hydroxytyrosol, and 3,4-dihydroxyphenylglycol are bioavailable and have anti-oxidant properties in vitamin E-deficient rats proteomics and network analysis approach. Mol Nutr Food Res 56:1137–1147CrossRefGoogle Scholar
  13. 13.
    Rubio-Senent F, Rodríguez-Gutiérrez G, Lama-Muñoz A, Fernández-Bolaños J (2013) Chemical characterization and properties of a polymeric phenolic fraction obtained from olive oil waste. Food Res Int 54:2122–2129CrossRefGoogle Scholar
  14. 14.
    Fernández-Bolaños J, Rodríguez G, Rodríguez R, Heredia A, Guillén R, Jiménez A (2002) Production in large quantities of highly purified hidroxitirosol from liquid–solid waste of two-phase olive oil processing or “Alperujo”. J Agric Food Chem 50:6804–6811CrossRefGoogle Scholar
  15. 15.
    Fernández-Bolaños J, Rodríguez G, Lama A, Rodríguez-Arcos R, Jiménez A, Guillén R (2008) Purification of 3,4-dihydroxyphenylglycol (DHPG) from vegetable products. Patent Request No 200803630Google Scholar
  16. 16.
    Fernández-Bolaños J, Rodríguez-Gutiérrez G, Lama-Muñoz A, Fernández-Bolaños JM, Maya-Castilla I, Rubio Senent F, López López O, Marset Castro A (2013) Method for obtaining hydroxytyrosol extract, mixture of hydroxytyrosol and 3,4-dihydroxyphenylglycol extract, and hydroxytyrosyl acetate extract from by-products of the olive tree and the purification of thereof. International Patent No: WO 2013/007850A1Google Scholar
  17. 17.
    Singleton VL, Rossi JA Jr (1965) Colorimetry of total phenolics with phosphomolybdic–phosphotungstic acid reagents. Am J Enol Vitic 16:144–158Google Scholar
  18. 18.
    Ostertag LM, O’Kennedy N, Horgan GW, Kroon PA, Duthie GG, de Roos B (2011) In vitro anti-platelet effects of simple plant-derived phenolic compounds are only found at high, non-physiological concentrations. Mol Nutr Food Res 55:1–13CrossRefGoogle Scholar
  19. 19.
    Duthie GG, González BM, Morrice PC, Arthur JR (1991) Inhibitory effects of isomers of tocopherol on lipid-peroxidation of microsomes from vitamin E-deficient rats. Free Radic Res Commun 15:35–40CrossRefGoogle Scholar
  20. 20.
    Burczynski JM, Southard SJ, Hayes JR, Longhurst PA, Colby HD (2001) Changes in mitochondrial and microsomal lipid peroxidation and fatty acid profiles in adrenal glands, testes, and livers from α-tocopherol-deficient rats. Free Radic Biol Med 30:1029–1035CrossRefGoogle Scholar
  21. 21.
    González-Correa JA, López-Villodres JA, Asensi R, Espartero JL, Rodríguez-Gutiérrez G, de la Cruz JP (2009) Virgen olive oil polyphenol hidroxitirosol acetate inhibits in vitro platelet aggregation in human whole blood: comparison with hidroxitirosol and acetylsalicylic acid. Br J Nutr 101:1157–1164CrossRefGoogle Scholar
  22. 22.
    Hubbard GP, Wolffram S, Lovegrove JA, Gibbins JM (2003) The role of polyphenols compounds in the diet as inhibitors of platelet function. Proc Nutr Soc 62:469–478CrossRefGoogle Scholar
  23. 23.
    Lill G, Voit S, Schror K, Weber AA (2003) Complex effect of different green tea catechins on human platelet. FEBS Lett 546:265–270CrossRefGoogle Scholar
  24. 24.
    Mitchell JH, Gardner PT, McPhail DB, Morrice PC, Collins AR, Duthie GG (1998) Anti-oxidant efficacy of phytoestrogens in chemical and biological model systems. Arch Biochem Biophys 360(1):142–148CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Fátima Rubio-Senent
    • 1
  • Baukje de Roos
    • 2
  • Garry Duthie
    • 2
  • Juan Fernández-Bolaños
    • 1
  • Guillermo Rodríguez-Gutiérrez
    • 1
    Email author
  1. 1.Food Phytochemistry DepartmentInstituto de la Grasa (CSIC)SevilleSpain
  2. 2.Rowett Institute of Nutrition and HealthUniversity of AberdeenAberdeenUK

Personalised recommendations