Skip to main content

Advertisement

SpringerLink
Log in

Qualitative screening of phenolic compounds in olive leaf extracts by hyphenated liquid chromatography and preliminary evaluation of cytotoxic activity against human breast cancer cells

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

In this work, high-performance liquid chromatography (HPLC) coupled to electrospray time-of-flight mass spectrometry (ESI-TOF-MS) and electrospray ion trap multiple-stage tandem mass spectrometry (ESI-IT-MS2) has been applied to screen phenolic compounds in olive leaf extracts. The use of a small particle size C18 column (1.8 μm) provided great resolution and made separation of a lot of isomers possible. The structural characterization was based on accurate mass data obtained by ESI-TOF-MS, and the nature of fragmentation ions were further confirmed by ESI-IT-MS2 when possible. In addition, we employed tetrazolium salt (MTT)-based assays to assess the effects of olive leaf extracts on the growth of human tumor-derived cells. Upon this approach, we achieved an accurate profile of olive leaf phenolics along with the identification of several important isomers of secoiridoids and flavonoids. This will allow a better understanding of the complete composition of olive-leaf-bioactive compounds as well as their involvement in Olea europaea L. biochemical pathways. Importantly, olive leaf extracts exhibited dose-dependent inhibitory effects on the metabolic status (cell viability) of three breast cancer models in vitro. Since the tumoricidal activity of the extracts should be mainly attributed to the identified olive leaf phenolics, these findings warrant further investigation at the structure–function molecular level to definitely establish the anticancer value of these phytochemicals.

Survival plots obtained by using the MTT assay for SKBr3, MCF-7 and JIMT-1 breast cancer cell lines treated with olive leaf extract at increasing concentrations for 72 h.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Estruch R, Martínez-González MA, Corella D, Salas-Salvadó J, Ruiz-Gutiérrez V, Covas MI, Fiol M, Gómez-Gracia E, López-Sabater MC, Vinyoles E, Arós F, Conde M, Lahoz C, Lapetra J, Sáez G, Ros E (2006) Ann Intern Med 145:1–11

    Google Scholar 

  2. Fitó M, Guxens M, Corella D, Sáez G, Estruch R, de la Torre R, Francés F, Cabezas C, López-Sabater MC, Marrugat J, García-Arellano A, Arós F, Ruiz-Gutierrez V, Ros E, Salas-Salvadó J, Fiol M, Solá R, Covas MI (2007) Arch Intern Med 167:1195–1203

    Article  Google Scholar 

  3. Scarmeas N, Stern Y, Tang MX, Mayeux R, Luchsinger JA (2006) Ann Neurol 59:912–921

    Article  Google Scholar 

  4. Trichopoulou A, Costacou T, Bamia C, Trichopoulos D (2003) New Engl J Med 348:2599–2608

    Article  Google Scholar 

  5. Menéndez JA, Vázquez-Martín A, Oliveras-Ferraros C, Garcia-Villalba R, Carrasco-Pancorbo A, Fernandez-Gutierrez A, Segura-Carretero A (2008) Int J Mol Med 22:433–439

    Google Scholar 

  6. Menéndez JA, Vázquez-Martín A, Colomer R, Brunet J, Carrasco-Pancorbo A, Garcia-Villalba R, Fernández-Gutiérrez A, Segura-Carretero A (2007) BMC Cancer 7:80–98

    Article  Google Scholar 

  7. Oliveras-López MJ, Innocenti M, Giaccherini C, Ieri F, Romani A, Mulinacci N (2007) Talanta 73:726–732

    Article  Google Scholar 

  8. Suárez M, Macià A, Romero MP, Motilva MJ (2008) J Chromatogr A 1214:90–99

    Article  Google Scholar 

  9. Obied HK, Bedgood DR, Prenzler PD, Robards K (2007) Anal Chim Acta 603:176–189

    Article  CAS  Google Scholar 

  10. Savarese M, Marco ED, Sacchi R (2007) Food Chem 105:761–770

    Article  CAS  Google Scholar 

  11. Obied HK, Allen MS, Bedgood DR, Prenzler PD, Robards K, Stockmann R (2005) J Agric Food Chem 53:823–837

    Article  CAS  Google Scholar 

  12. Donna LD, Mazzotti F, Salerno R, Tagarelli A, Taverna D, Sindona G (2007) Rapid Commun Mass Spectrom 21:3653–3657

    Article  Google Scholar 

  13. Arráez-Román D, Sawalha S, Segura-Carretero A, Menéndez JA, Fernández-Gutiérrez A (2008) Agro Food Ind Hi Tec 19:18–22

    Google Scholar 

  14. Paiva-Martins F, Pinto M (2008) J Agric Food Chem 56:5582–5588

    Article  CAS  Google Scholar 

  15. Food and Agriculture Organization of the United Nations, Database of the Statistics Division. http://faostat.fao.org

  16. Molina-Alcaidea E, Nefzaouib A (1996) Int Biodeter Biodegr 38:227–235

    Article  Google Scholar 

  17. Perrinjaquet-Moccetti T, Busjahn A, Schmidlin C, Schmidt A, Bradl B, Aydogan C (2008) Phytother Res 22:1239–1242

    Article  Google Scholar 

  18. Zarzuelo A, Duarte J, Jiménez J, González M, Utrilla MP (1991) Planta Med 57:417–419

    Article  CAS  Google Scholar 

  19. Fredrickson WR. US patent 2000, No. 6117844

  20. Micol V, Caturla N, Pérez-Fons L, Más V, Pérez L, Estepa A (2005) Antivir Res 66:129–136

    Article  CAS  Google Scholar 

  21. Cavaliere C, Foglia P, Gubbiotti R, Sacchetti P, Samperi R, Laganá A (2008) Rapid Commun Mass Spectrom 22:3089–3099

    Article  CAS  Google Scholar 

  22. Ferrer I, Thurman EM, Zwelgenbaum JA (2008) Rapid Commun Mass Spectrom 21:3869–3882

    Article  Google Scholar 

  23. Fu S, Arráez-Román D, Menéndez JA, Segura-Carretero A, Fernández-Gutiérrez A (2009) Rapid Commun Mass Spectrom 23:51–59

    Article  Google Scholar 

  24. Cardoso SM, Guyot S, Marnet N, Lopes-da-Silva JA, Renard CMGC, Coimbra MA (2005) J Sci Food Agric 85:21–32

    Article  CAS  Google Scholar 

  25. De Nino A, Lombardo N, Perri E, Procopio A, Raffaelli A, Sindona G (1997) J Mass Spectrom 32:533–541

    Article  Google Scholar 

  26. Ryan D, Robards K, Prenzler P, Jardine D, Herlt T, Antolovich M (1999) J Chromatogr A 855:529–537

    Article  CAS  Google Scholar 

  27. Ryan D, Antolovich M, Herlt T, Prenzler PD, Lavee S, Robards K (2002) J Agric Food Chem 50:6716–6724

    Article  CAS  Google Scholar 

  28. Obied HK, Prenzler PD, Ryan D, Servili M, Taticchi A, Espostob S, Robards K (2008) Nat Prod Rep 25:1167–1179

    Article  CAS  Google Scholar 

  29. Torre-Carbot K, Jauregui O, Gimeno E, Castellote AI, Lamuela-Raventós RM, López-Sabater MC (2005) J Agric Food Chem 53:4331–4340

    Article  Google Scholar 

  30. Nino AD, Mazzotti F, Morrone SP, Perri E, Raffaelli A, Sindona G (1999) J Mass Spectrom 34:10–16

    Article  Google Scholar 

  31. Karabourniotis G, Papadopoulos K, Papamarkou M, Manetas Y (1992) Physiol Plantarum 86:414–418

    Article  Google Scholar 

  32. Sánchez-Ávila N, Priego-Capote F, Ruiz-Jiménez J, Ruiz-Jiménez J, Luque de Castro MD (2009) Talanta 78:40–48

    Article  Google Scholar 

  33. Kaefer CM, Milner JA (2008) J Nutr Biochem 19:347–361

    Article  CAS  Google Scholar 

  34. Shoemaker M, Hamilton B, Dairkee SH, Cohen I, Campbell MJ (2005) Phytother Res 19:649–651

    Article  Google Scholar 

  35. Friedman M, Mackey BE, Kim HJ, Lee IS, Lee KR, Lee SU, Kozukue E, Kozukue N (2007) J Agric Food Chem 55:243–253

    Article  CAS  Google Scholar 

  36. Barrajón-Catalán E, Menéndez-Gutiérrez MP, Falco A, Carrato A, Saceda M, Micol V (2010) Cancer Lett 290:192–203

    Google Scholar 

  37. Caruso D, Colombo R, Patelli R, Giavarini F, Galli G (2000) J Agric Food Chem 48:1182–1185

    Article  CAS  Google Scholar 

  38. Japón-Luján R, Castro MDL (2007) J Agric Food Chem 55:4584–4588

    Article  Google Scholar 

  39. Altıok E, Bayçın D, Bayraktar O, Ülkü S (2008) Sep Purif Technol 62:342–348

    Article  Google Scholar 

  40. Christophoridou S, Dais P, Tseng LH, Spraul M (2005) J Agric Food Chem 53:4667–4679

    Article  CAS  Google Scholar 

  41. Bianchi G (2003) Eur J Lipid Sci Tech 105:229–242

    Article  CAS  Google Scholar 

  42. Sánchez-Ávila N, Priego-Capote F, Ruiz-Jiménez J, Castro MDL (2009) Talanta 78:40–48

    Article  Google Scholar 

  43. García-Villalba R, León C, Dinelli G, Segura-Carretero A, Fernández-Gutiérrez A, Garcia-Caňas V, Cifuentes A (2008) J Chromatogr A 1195:164–173

    Article  Google Scholar 

Download references

Acknowledgments

The author, Dr. Shaoping Fu, acknowledges financial support from the Spanish Ministry of Education and Science (project SB2006-0157). The author, Dr. David Arráez-Román, also acknowledges financial support from the Consejería de Innovación, Ciencia y Empresa (Junta de Andalucía, Excellence Project P07-AGR-02619, Contrato reincorporación de doctores). The authors also gratefully acknowledge the financial support from MEC (AGL2008-05108-CO3-03/ALI and AGL2007-60778/ALI to V. Micol) and Junta de Andalucía, Excellence Project P09-CTS-4564. Work by Javier A. Menendez was supported in part by Instituto de Salud Carlos III (Ministerio de Sanidad y Consumo, Fondo de Investigación Sanitaria-FIS, Spain, Grants CP05-00090, PI06-0778 and RD06-0020-0028 to Javier A. Menendez). Javier A. Menendez was also supported by a Grant from the Fundación Científica de la Asociación Española Contra el Cáncer (AECC, Spain). We also thank New Developments in Nutraceuticals, Inc., for providing us with the olive leaf extracts.

Author information

Author notes

  1. María P. Menéndez-Gutiérrez

    Present address: Regenerative Cardiology Department, National Center for Cardiovascular Research Foundation, C/ Melchor Fernández Almagro, 3, 28029, Madrid, Spain

Authors and Affiliations

  1. Institute of Chemistry and Applications of Plant Resources, School of Biological & Food Engineering, Dalian Polytechnic University, 1 Qinggong Yuan, Ganjingzi District, Dalian, 116034, China

    Shaoping Fu

  2. Department of Analytical Chemistry, Faculty of Sciences, University of Granada, C/Fuentenueva, 18071, Granada, Spain

    Shaoping Fu, David Arráez-Roman, Antonio Segura-Carretero & Alberto Fernández-Gutiérrez

  3. Institute of Molecular & Cell Biology, University Miguel Hernández, 03202, Elche, Alicante, Spain

    Vicente Micol

  4. Girona Biomedical Research Institute (IdIBGi), Catalan Institute of Oncology, Girona (ICO-Girona), 17007, Girona, Spain

    Javier A. Menéndez

Authors
  1. Shaoping Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David Arráez-Roman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Antonio Segura-Carretero
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Javier A. Menéndez
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. María P. Menéndez-Gutiérrez
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Vicente Micol
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Alberto Fernández-Gutiérrez
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Antonio Segura-Carretero.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 500 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fu, S., Arráez-Roman, D., Segura-Carretero, A. et al. Qualitative screening of phenolic compounds in olive leaf extracts by hyphenated liquid chromatography and preliminary evaluation of cytotoxic activity against human breast cancer cells. Anal Bioanal Chem 397, 643–654 (2010). https://doi.org/10.1007/s00216-010-3604-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-010-3604-0

Keywords

Search

Navigation