Micro-scaled Quantitative Method to Analyze Olive Oil Polyphenols

  • Giovanni Caprioli
  • Maria Chiara Boarelli
  • Massimo Ricciutelli
  • Gianni Sagratini
  • Dennis FioriniEmail author


This study aims to improve an analytical method to quantify phenolic substances in olive oil. In order to minimize time required and quantity of solvents, sample extract preparation performed for a previously developed high-performance liquid chromatography–diode array detector to quantify olive oil polyphenols has been ten times downscaled and then validated. The new method performs the extraction of phenolic substances from 0.5 g of oil and allows to quantify the phenolic acids vanillic acid, p-coumaric acid, and ferulic acid; the phenolic alcohols tyrosol and hydroxytyrosol; secoiridoid derivatives; the lignans pinoresinol and acetoxypinoresinol; and the flavonoids luteolin and apigenin. Recoveries obtained were 66–89% for phenolic alcohols, 64–90% for phenolic acids, 93–96% for oleuropein (used as a reference for secoiridoid derivatives), 71–95% for flavonoids, and 97–100% for lignans. The total quantity of organic solvents used in the sample preparation is decreased from 30 to 3 mL with an important abatement of waste, costs, and working time requested.


Olive oil polyphenols Quantitative determination HPLC-DAD Secoiridoid derivatives Green method 



This study was funded by the University of Camerino (grant “Fondo di Ateneo per la Ricerca” 2014–2015, project: “AEVOO: Authentication of Extra Virgin Olive Oil”).

Compliance with Ethical Standards

Conflict of Interest

Giovanni Caprioli declares that he has no conflict of interest. Maria Chiara Boarelli declares that she has no conflict of interest. Massimo Ricciutelli declares that he has no conflict of interest. Gianni Sagratini declares that he has no conflict of interest. Dennis Fiorini declares that she has no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Not applicable.


  1. Alessandri S, Ieri F, Romani A (2014) Minor polar compounds in extra virgin olive oil: correlation between HPLC-DAD-MS and the Folin-Ciocalteu spectrophotometric method. J Agric Food Chem 62:826–835CrossRefGoogle Scholar
  2. Bakhouche A, Lozano-Sanchez J, Beltran-Debon R, Joven J, Segura-Carretero A, Fernandez-Gutierrez A (2013) Phenolic characterization and geographical classification of commercial Arbequina extra-virgin olive oils produced in southern Catalonia. Food Res Int 50:401–408CrossRefGoogle Scholar
  3. Bayram B, Esatbeyoglu T, Schulze N, Ozcelik B, Frank J, Rimbach G (2012) Comprehensive analysis of polyphenols in 55 extra virgin olive oils by HPLC-ECD and their correlation with antioxidant activities. Plant Foods Hum Nutr 67:326–336CrossRefGoogle Scholar
  4. Bendini A, Cerretani L, Carrasco-Pancorbo A, Gómez-Caravaca AM, Segura-Carretero A, Fernández-Gutiérrez A, Lercker G (2007) Phenolic molecules in virgin olive oils: a survey of their sensory properties, health effects, antioxidant activity and analytical methods. An overview of the last decade. Molecules 12:1679–1719CrossRefGoogle Scholar
  5. Caporaso N, Savarese M, Paduano A, Guidone G, De Marco E, Sacchi R (2015) Nutritional quality assessment of extra virgin olive oil from the Italian retail market: do natural antioxidants satisfy EFSA health claims? J Food Compos Anal 40:154–162CrossRefGoogle Scholar
  6. Capriotti AL, Cavaliere C, Crescenzi C, Foglia P, Nascatelli R, Samperi R, Laganà A (2014) Comparison of extraction methods for the identification and quantification of polyphenols in virgin olive oil by ultra-HPLC-QtoF mass spectrometry. Food Chem 158:392–400CrossRefGoogle Scholar
  7. Carrasco-Pancorbo A, Cerretani L, Bendini A, Segura-Carretero A, Gallina-Toschi T, Fernandez-Gutiérrez A (2005) Analytical determination of polyphenols in olive oils. J Sep Sci 28:837–858CrossRefGoogle Scholar
  8. De La Torre-Carbot J, Gimeno C, Raventos LS (2005) Characterization and quantification of phenolic compounds in olive oils by solid-phase extraction, HPLC-DAD, and HPLC-MS/MS. J Agric Food Chem 53:4331–4340CrossRefGoogle Scholar
  9. EU (2012) Establishing a list of permitted health claims made on foods, other than those referring to the reduction of disease risk and to children’s development and health. Commission Regulation (EU) 432/2012. Off J Eur Union 136:1–40Google Scholar
  10. Flores MIA, Romero-González R, Frenich AG, Vidal JLM (2012) Analysis of phenolic compounds in olive oil by solid-phase extraction and ultra high performance liquid chromatography-tandem mass spectrometry. Food Chem 134:2465–2472CrossRefGoogle Scholar
  11. Franco MN, Galeano-Diaz T, Lopez O, Fernandez-Bolanos JG, Sanchez J, De MC, Gil MV, Martin-Vertedor D (2014) Phenolic compounds and antioxidant capacity of virgin olive oil. Food Chem 163:289–298CrossRefGoogle Scholar
  12. IOC (2009) Determination of biophenols in olive oils by HPLC. COI/T.20/Doc. No 29/1–8Google Scholar
  13. Montedoro GF, Servili M, Baldioli M, Miniati E (1992) Simple and hydrolyzable phenolic compounds in virgin olive oil. 1. Their extraction, separation, and quantitative and semiquantitative evaluation by HPLC. J Agric Food Chem 40:1571–1576CrossRefGoogle Scholar
  14. Pirisi FM, Angioni A, Cabras P, Garau VL, Di Teulada MTS, Dos Santos MK, Bandino G (1997) Phenolic compounds in virgin olive oils. I. Low-wavelength quantitative determination of complex phenols by high-performance liquid chromatography under isocratic elution. J Chromatogr A 768:207–213CrossRefGoogle Scholar
  15. Pirisi FM, Cabras P, Falqui Cao C, Migliorini M, Muggelli M (2000) Phenolic compounds in virgin olive oil. 2. Reappraisal of the extraction, HPLC separation, and quantification procedures. J Agric Food Chem 48:1191–1196CrossRefGoogle Scholar
  16. Ricciutelli M, Marconi S, Boarelli MC, Caprioli G, Sagratini G, Ballini R, Fiorini D (2017) Olive oil polyphenols: a quantitative method by high-performance liquid-chromatography-diode-array detection for their determination and the assessment of the related health claim. J Chromatogr A 1481:53–63CrossRefGoogle Scholar
  17. Selvaggini R, Servili M, Urbani S, Esposto S, Taticchi A, Montedoro G (2006) Evaluation of phenolic compounds in virgin olive oil by direct injection in high-performance liquid chromatography with fluorimetric detection. J Agric Food Chem 54:2832–2838CrossRefGoogle Scholar
  18. Servili M, Sordini B, Esposto S, Urbani S, Veneziani G, Di Maio I, Selvaggini R, Taticchi A (2014) Biological activities of phenolic compounds of extra virgin olive oil. Antioxidants 3:1–23CrossRefGoogle Scholar
  19. Suárez M, Macià A, Romero MP, Motilva MJ (2008) Improved liquid chromatography tandem mass spectrometry method for the determination of phenolic compounds in virgin olive oil. J Chromatogr A 1214:90–99CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of PharmacyUniversity of CamerinoCamerinoItaly
  2. 2.School of Science and Technology, Chemistry DivisionUniversity of CamerinoCamerinoItaly
  3. 3.HPLC-MS LaboratoryUniversity of CamerinoCamerinoItaly

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