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Open AccessFeature PaperArticle

In House Validated UHPLC Protocol for the Determination of the Total Hydroxytyrosol and Tyrosol Content in Virgin Olive Oil Fit for the Purpose of the Health Claim Introduced by the EC Regulation 432/2012 for “Olive Oil Polyphenols”

1
Laboratory of Food Chemistry and Technology, School of Chemistry, Aristotle University of Thessaloniki (AUTH), 541 24 Thessaloniki, Greece
2
Instituto de la Grasa (CSIC), Ctra. de Utrera, km. 1, Campus Universitario Pablo de Olavide-Building 46, 41013 Seville, Spain
3
Department of Agricultural and Food Sciences, Alma Mater Studiorum-University of Bologna (UNIBO), Piazza Goidanich, 60, I-47521 Cesena (FC), Italy
*
Author to whom correspondence should be addressed.
Molecules 2019, 24(6), 1044; https://doi.org/10.3390/molecules24061044
Received: 25 February 2019 / Revised: 12 March 2019 / Accepted: 12 March 2019 / Published: 16 March 2019
(This article belongs to the Special Issue Olive Bioactives: From Molecules to Human Health)
An ongoing challenge in olive oil analytics is the development of a reliable procedure that can draw the consensus of all interested parties regarding the quantification of concentrations above the required minimum value of 5 mg of bioactive “olive oil polyphenols” per 20 g of the oil, to fulfill the health claim introduced by the European Commission (EC) Regulation 432/2012. An in-house validated ultra-high performance liquid chromatography (UHPLC) protocol fit for this purpose is proposed. It relies on quantification of the total hydroxytyrsol (Htyr) and tyrosol (Tyr) content in the virgin olive oil (VOO) polar fraction (PF) before and after acidic hydrolysis of their bound forms. PF extraction and hydrolysis conditions were as previously reported. The chromatographic run lasts ~1/3 of the time needed under high performance liquid chromatography (HPLC) conditions, this was also examined. Eluent consumption for the same piece of information was 6-fold less. Apart from being cost effective, a larger number of samples can be analyzed daily with less environmental impact. Two external curves, detection at 280 nm and correction factors for molecular weight difference are proposed. The method, which is fit for purpose, is selective, robust with satisfactory precision (percentage relative standard deviation (%RSD) values < 11%) and recoveries higher than 87.6% for the target analytes (Htyr, Tyr). Standard operational procedures are easy to apply in the olive oil sector. View Full-Text
Keywords: UHPLC; diode array detection; hydroxytyrosol; tyrosol; health claim; virgin olive oil; validation; European Commission Regulation 432/2012; olive oil phenols UHPLC; diode array detection; hydroxytyrosol; tyrosol; health claim; virgin olive oil; validation; European Commission Regulation 432/2012; olive oil phenols
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Tsimidou, M.Z.; Sotiroglou, M.; Mastralexi, A.; Nenadis, N.; García-González, D.L.; Gallina Toschi, T. In House Validated UHPLC Protocol for the Determination of the Total Hydroxytyrosol and Tyrosol Content in Virgin Olive Oil Fit for the Purpose of the Health Claim Introduced by the EC Regulation 432/2012 for “Olive Oil Polyphenols”. Molecules 2019, 24, 1044.

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