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Toward a Harmonized and Standardized Protocol for the Determination of Total Hydroxytyrosol and Tyrosol Content in Virgin Olive Oil (VOO). The Pros of a Fit for the Purpose Ultra High Performance Liquid Chromatography (UHPLC) Procedure

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Laboratory of Food Chemistry and Technology, School of Chemistry, Aristotle University of Thessaloniki (AUTH), 541 24 Thessaloniki, Greece
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Department of Agricultural, Food and Environmental Sciences, University of Perugia, (UNIPG), Via San Costanzo s.n.c., 06126 Perugia, Italy
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Laboratory of the Institute for Oliveculture, Science and Research Centre Koper (ZRS), Zelena ulica 8k, SI-6310 Izola, Slovenia
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Department de Nutrició, Ciències de l’Alimentació i Gastronomia, XaRTA, INSA-UB, Campus De l’Alimentació Torribera, Universitat de Barcelona (UB), Av. Prat de la Riba, 171, 08028 Santa Coloma de Gramenet, Spain
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Eurofins Analytik GmbH, Neuländer Kamp 1, 21079 Hamburg, Germany
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Instituto de la Grasa (CSIC), Ctra. De Utrera, km. 1, Campus Universitario Pablo de Olavide—Building 46, 41013 Seville, Spain
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Department of Agricultural and Food Sciences, Alma Mater Studiorum, University of Bologna (UNIBO), Piazza Goidanich, 60, I-47521 Cesena (FC), Bologna, Italy
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Author to whom correspondence should be addressed.
Molecules 2019, 24(13), 2429; https://doi.org/10.3390/molecules24132429
Received: 28 May 2019 / Revised: 24 June 2019 / Accepted: 28 June 2019 / Published: 2 July 2019
(This article belongs to the Special Issue Olive Bioactives: From Molecules to Human Health)
Τoward a harmonized and standardized procedure for the determination of total hydroxytyrosol and tyrosol content in virgin olive oil (VOO), the pros of a recently published in house validated ultra high performance liquid chromatography (UHPLC) protocol are discussed comparatively with those of other procedures that determine directly or indirectly the compounds hosted under the health claim on “olive oil polyphenols” (EC regulation 432/2012). Authentic VOOs were analyzed with five different liquid chromatographic separation protocols and 1H-NMR one in five different laboratories with expertise in VOO phenol analysis within three months. Data comparison indicated differences in absolute values. Method comparison using appropriate tools (Passing-Bablok regression and Bland Altman analyses) for all protocols vs. the UHPLC one indicated slight or statistically significant differences. The results were also discussed in terms of cost effectiveness, detection means, standard requirements and ways to calculate the total hydroxytyrosol and tyrosol content. Findings point out that the in-house validated fit for the purpose UHPLC protocol presents certain pros that should be exploited by the interested parties. These are the simplicity of sample preparation, fast elution time that increase the number of samples analyzed per day and integration of well-resolved peaks with the aid of only two commercially available external standards. Importance of correction factors in the calculations is stressed. View Full-Text
Keywords: virgin olive oil; hydroxytyrosol; tyrosol; health claim; European Commission Regulation 432/2012; standardization; UHPLC-DAD; HPLC-DAD; LC-HRMS; 1H-NMR spectroscopy virgin olive oil; hydroxytyrosol; tyrosol; health claim; European Commission Regulation 432/2012; standardization; UHPLC-DAD; HPLC-DAD; LC-HRMS; 1H-NMR spectroscopy
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Tsimidou, M.Z.; Nenadis, N.; Mastralexi, A.; Servili, M.; Butinar, B.; Vichi, S.; Winkelmann, O.; García-González, D.L.; Toschi, T.G. Toward a Harmonized and Standardized Protocol for the Determination of Total Hydroxytyrosol and Tyrosol Content in Virgin Olive Oil (VOO). The Pros of a Fit for the Purpose Ultra High Performance Liquid Chromatography (UHPLC) Procedure. Molecules 2019, 24, 2429.

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