Numerous studies have been carried out on the bioactive properties of hydroxytyrosol (HT) in olive oils (OOs), although there are few reports comparing anti-inflammatory activity among different olive varieties or regions of production. The purpose of this study was to investigate the in vitro inflammatory action of HT in extracts of four OO varieties in the Languedoc region of the French Mediterranean. Factors other than cultivar were eliminated, which enabled unambiguous varietal differences to be identified. Purified extracts of OO were obtained using an optimized solid-phase extraction procedure by which only polar compounds were recovered. High performance liquid chromatography-photodiode array detection-tandem mass spectrometry was used to identify and quantify HT and oleacein in the extracts. The total polyphenol concentration ranged from 93.00 mg gallic acid equivalent/kg OO for Picholine to 27 mg gallic acid equivalent for Verdale OOs. The concentrations of HT in Picholine, Olivère, and Lucques varieties were 25.3, 18.8 and 12.1 mg/kg, respectively, whereas the concentration of HT in Verdale OOs was less, 1 mg/kg. The in vitro anti-inflammatory response of purified OO extracts, evaluated by the inhibition of nitric oxide release in lipopolysaccharide-induced interferon-γ activated J774.A1 macrophages, strongly correlated with total polyphenol content (R2 > 0.995). The effect increased asymptotically between the equivalent of 2 and 37 mg of OO, reaching, at the maximum tested concentration, 90%, 75%, 62%, and 30% activity for Picholine, Olivière, Lucques, and Verdale, respectively. The results presented here clearly show that, by comparison with authentic standards, the activity of HT in OO extracts was enhanced in a concentration-dependent manner, varying from 3-fold at the highest extract concentration to over 6.5-fold at the lowest extract concentration. Therefore, the anti-inflammatory activity of OOs should be rationalized on the basis of whole extracts rather than solely on the concentration of HT or other bioactive compounds in OO.
Keywords: LPS-activated J774.A1 macrophages; NO release; Southern France olive oils and olive oil extracts; anti-inflammatory; hydroxytyrosol.