Abstract

Virgin olive oil (VOO) is one of the most emblematic products of the Mediterranean diet. Its content in phenolic compounds is strongly associated with the antioxidant and health-promoting properties of this diet. VOO’s phenolic profile is determined mainly by the phenolic compounds present in the olive fruit, so knowing their content allows for a fairly precise estimate of the antioxidant and functional properties of the corresponding oil. In this sense, a convenient, green, and sensitive spectrophotometric method was developed for the quantitative determination of total phenolic compounds in olive fruits. The method is based on an easy-to-use extraction procedure of olive fruit phenolics using dimethyl sulfoxide and quantification with the Folin–Ciocalteu reagent. Oleuropein proved to be a suitable reference compound for quantification, displaying a good linear response (r = 0.9996) over the concentration range of 0.58–6.48 mg/mL, with a variation coefficient of 0.42% and limits of detection and quantification of 0.0492 and 0.1490 mg/mL, respectively. The method was validated using a wide array of fruit samples representative of the Olea europaea L. genetic diversity. The results obtained with this spectrophotometric method, expressed as mg/mL of oleuropein, showed a good correlation with those obtained with the fruit samples analyzed by high performance liquid chromatography, with an r value of 0.9930 and a slope value of 1.022, confirming its reliability. Thus, this method can become a very useful simple tool to estimate the total phenolic content of olive fruits, especially when working with numerous samples such as in olive breeding programs or in commercial olive production, in which it is especially useful to know the phenolic state of the fruit and thus determine the optimal harvest date or the most appropriate agronomic treatment to increase the functional properties of the olive fruit and the olive oil.

Highlights

  • We developed a fast, reliable, and flexible procedure for the extraction of the main phenolic compounds in olive fruit based on the use of Dimethyl sulfoxide (DMSO) as an extractant agent, which through subsequent high performance liquid chromatography (HPLC) analysis allows us to know the phenolic profile of this fruit [8]

  • Ligstroside and demethyloleuropein, the major phenolic glucosides in olive fruit, and others such as verbascoside and the flavonoids rutin, luteolin, and apigenin glucosides are present in significant amounts [11]

  • Anthocyanins are synthetized during fruit ripening and constitute a major class of compounds within the olive fruit phenolics

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Summary

Introduction

The phenolics of virgin olive oil (VOO) have attracted special attention in recent years for their proven properties that promote human health, regarding cardiovascular diseases, inflammation, or cancer [1,2]. A health claim on the evidence of the protective effect of VOO phenolics for cardiovascular diseases was approved by the European Food Safety Authority [3], which applies to those oils that contain at least 250 ppm of hydroxytyrosol and derivatives. The phenolic fraction of VOO is made up of an array of compounds belonging to different chemical classes such as secoiridoids, lignans, flavonoids, a series of alcohols, and simple phenolic acids, with the main phenolic derivatives of oil being the secoiridoids. The presence of secoiridoids in VOO is related to the content of phenolic glycosides with the secoiridoid structure present in the olive fruit, whose main representatives are oleuropein, ligstroside, and demethyloleuropein, and the activity of the hydrolytic and oxidative enzymes that

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