Comparative elucidation of phenolic compounds in Albanian olive oils using LC-DAD-ESI-MS/MS

Abstract

Olive oils may provide health benefits, including the prevention of coronary heart diseases, cancers, and the modification of immune and inflammatory responses. These benefits mainly originate from the phenolic compounds found in olive oil. There has been no study on the advanced characterization of Albanian olive oils from various cultivars regarding phenolic compounds. Hence, a comprehensive characterization of phenolic compounds is carried out in Albanian monocultivar virgin olive oils from five different cultivars, including Kalinjot, Bardhi Tirana, Ulliri-i-Zi Tirana, Krips Kruja, and Bardhi Kruja for the first time. Liquid chromatography coupled to diode array detection and electrospray ?onization tandem mass spectrometry (LC-DAD-ESI-MS/MS) is employed for the determination of phenolic compounds. In total, 18 compounds were identified in all samples, including phenolic alcohols, phenolic acids, secoiridoids, flavonoids, and phenolic aldehydes. Significant quantitative differences were detected among the cultivars, with the highest concentrations detected in virgin olive oil (VOO) from cv. Ulli-i-Zi. Secoiridoids were found in abundance, in general, followed by phenolic alcohols, and in this group, 3,4-DHPEA-EDA and p-HPEA-EDA stood out as dominant compounds, especially in Kalinjot virgin olive oils. Regarding phenolic alcohols, 3,4-DHPEA-AC was determined as the main phenolic compound. Phenolic profiles were found to be significantly different among the olive oil samples of different cultivars. Principal component analyses (PCA) displayed the differentiation of samples in terms of phenolic compounds.     

On the Squalene Content of CV Chondrolia Chalkidikis and Chalkidiki (Greece) Virgin Olive Oil

This work is a continuation of efforts to establish the nutritional profile of virgin olive oil (VOO) from cv. Chondrolia Chalkidikis and Chalkidiki and to strengthen its positioning in the global VOO landscape. VOOs produced at an industrial scale in different olive mills of the Chalkidiki (Greece) regional unit as well as VOOs obtained at the laboratory scale from drupes of different maturity stages for four consecutive harvesting years were examined for their squalene (SQ) content using both HPLC and GC procedures. The mean values of SQ were found to be 4228 (HPLC) and 4865 (GC) mg/kg oil (n = 15) and were of the same magnitude as that in VOOs from cv Koroneiki (4134 mg/kg, n = 23). Storage of VOOs in the dark at room temperature for 18 months indicated an insignificant mean SQ content loss (~2%) in comparison to a mean loss of 26% for alpha-tocopherol content. This finding strengthens our view that SQ does not act as a radical scavenger that donates hydrogen atoms to the latter. The four consecutive harvest years studied indicated a clear declining trend in VOO SQ concentration upon olive ripening. To our knowledge, this is the first systematic work concerning the SQ content of Chondrolia Chalkidikis and Chalkidiki VOOs.     

Comparative Assessment of Phytochemical Compounds and Antioxidant Properties of Kernel Oil from Eight Sour Cherry (Prunus cerasus L.) Cultivars

New plant oils as a potential natural source of nutraceutical compounds are still being sought. The main components of eight cultivars (‘Koral’, ‘Lucyna’, ‘Montmorency’, ‘Naumburger’, ‘Wanda’, ‘Wigor’, ‘Wołyńska’, and ‘Wróble’) of sour cherry (Prunus cerasus L.) grown in Poland, including crude fat, protein, and oil content, were evaluated. The extracted oils were analysed for chemical and biological activity. The oils had an average peroxide value of 1.49 mEq O₂/kg, acid value of 1.20 mg KOH/g, a saponification value of 184 mg of KOH/g, and iodine value of 120 g I₂/100 g of oil. The sour cherry oil contained linoleic (39.1–46.2%) and oleic (25.4–41.0%) acids as the major components with smaller concentrations of α-eleostearic acid (8.00–15.62%), palmitic acid (5.45–7.41%), and stearic acid (2.49–3.17%). The content of sterols and squalene varied significantly in all the studied cultivars and ranged between 336–973 mg/100 g and 66–102 mg/100 g of oil. The contents of total tocochromanols, polyphenols, and carotenoids were 119–164, 19.6–29.5, and 0.56–1.61 mg/100 g oil, respectively. The cultivar providing the highest amounts of oil and characterised by the highest content of PUFA (including linoleic acid), plant sterols, α-and β-tocopherol, as well as the highest total polyphenol and total carotenoids content was been found to be ‘Naumburger’. The antioxidant capacity of sour cherry kernel oils, measured using the DPPH^• and ABTS^•+ methods, ranged from 57.7 to 63.5 and from 38.2 to 43.2 mg trolox/100 g oil, respectively. The results of the present study provide important information about potential possibilities of application of Prunus cerasus kernel oils in cosmetic products and pharmaceuticals offering health benefits.     

Epigenetic Modifications Induced by Olive Oil and Its Phenolic Compounds: A Systematic Review

Many studies demonstrated that olive oil (especially extra virgin olive oil: EVOO) phenolic compounds are bioactive molecules with anti-cancer, anti-inflammatory, anti-aging and neuroprotective activities. These effects have been recently attributed to the ability of these compounds to induce epigenetics modifications such as miRNAs expression, DNA methylation and histone modifications. In this study, we systematically review and discuss, following the PRISMA statements, the epigenetic modifications induced by EVOO and its phenols in different experimental systems. At the end of literature search through “PubMed”, “Web of Science” and “Scopus”, 43 studies were selected.Among them, 22 studies reported data on miRNAs, 15 on DNA methylation and 13 on histone modification. Most of the “epigenomic” changes observed in response to olive oil phenols’ exposure were mechanistically associated with the cancer preventive and anti-inflammatory effects. In many cases, the epigenetics effects regarding the DNA methylation were demonstrated for olive oil but without any indication regarding the presence or not of phenols. Overall, the findings of the present systematic review may have important implications for understanding the epigenetic mechanisms behind the health effects of olive oil. However, generally no direct evidence was provided for the causal relationships between epigenetics modification and EVOO health related effects. Further studies are necessary to demonstrate the real physiological consequences of the epigenetics modification induced by EVOO and its phenolic compounds.     

Effects of Enzymatic Pretreatment of Seeds on the Physicochemical Properties,Bioactive Compounds,and Antioxidant Activity of Pomegranate Seed Oil

Enzymatic pretreatment of seeds is a novel approach that enhances the health benefits of the extracted oil. The study investigated the influence of the enzymatic pretreatment of seeds on the quality of oil from different pomegranate cultivars. The quality of the ultrasound-assisted (and ethanol-extracted) oil was studied, with respect to the refractive index (RI), yellowness index (YI), conjugated dienes (K232), peroxide value (PV) ρ-anisidine value (AV), total oxidation value (TOTOX), total carotenoid content (TCC), total phenolic compounds (TPC), fatty acid composition, phytosterol composition, ferric reducing antioxidant power (FRAP), and 2.2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging capacity. The seeds of three different pomegranate cultivars (‘Wonderful’, ‘Herskawitz’, and ‘Acco’) were digested with an equal mixture of Pectinex Ultra SPL, Flavourzyme 100 L, and cellulase crude enzymes, at a concentration, pH, temperature, and time of 1.7%, 4.5, 40 °C, and 5 h, respectively. Enzymatic pretreatment of PS increased oil yield, PV, TPC, TCC, and DPPH radical scavenging capacity, but decreased the YI. The levels of K232, AV and TOTOX, fatty acids, phytosterols, RI, and FRAP, were not significantly affected by enzymatic pretreatment of PS. Principal component analysis (PCA) established that oil extracted from the ‘Acco’ seed after enzymatic pretreatment had higher yield, TPC, TCC, and DPPH radical scavenging capacity. Therefore, enzyme-pretreated ‘Acco’ pomegranate fruit seed is a source of quality seed oil with excellent antioxidant properties.     

A New Definition of the Term “High-Phenolic Olive Oil” Based on Large Scale Statistical Data of Greek Olive Oils Analyzed by qNMR

In the last few years, a new term, “High-phenolic olive oil”, has appeared in scientific literature and in the market. However, there is no available definition of that term regarding the concentration limits of the phenolic ingredients of olive oil. For this purpose, we performed a large-scale screening and statistical evaluation of 5764 olive oil samples from Greece coming from >30 varieties for an eleven-year period with precisely measured phenolic content by qNMR. Although there is a large variation among the different cultivars, the mean concentration of total phenolic content was 483 mg/kg. The maximum concentration recorded in Greece reached 4003 mg/kg. We also observed a statistically significant correlation of the phenolic content with the harvest period and we also identified varieties affording olive oils with higher phenolic content. In addition, we performed a study of phenolic content loss during usual storage and we found an average loss of 46% in 12 months. We propose that the term high-phenolic should be used for olive oils with phenolic content > 500 mg/kg that will be able to retain the health claim limit (250 mg/kg) for at least 12 months after bottling. The term exceptionally high phenolic olive oil should be used for olive oil with phenolic content > 1200 mg/kg (top 5%).     

Processing Effect and Characterization of Olive Oils from Spanish Wild Olive Trees (Olea europaea var. sylvestris)

Wild olive trees have important potential, but, to date, the oil from wild olives has not been studied significantly, especially from an analytical point of view. In Spain, the wild olive tree is called “Acebuche” and its fruit “Acebuchina”. The objective of this work is to optimize the olive oil production process from the Acebuchina cultivar and characterize the oil, which could be marketed as healthy and functional food. A Box–Behnken experimental design with five central points was used, along with the Response Surface Methodology to obtain a mathematical experimental model. The oils from the Acebuchina cultivar meet the requirements for human consumption and have a good balance of fatty acids. In addition, the oils are rich in antioxidants and volatile compounds. The highest extraction yield, 12.0 g oil/100 g paste, was obtained at 90.0 min and the highest yield of phenolic compounds, 870.0 mg/kg, was achieved at 40.0 °C, and 90.0 min; but the maximum content of volatile compounds, 26.9 mg/kg, was obtained at 20 °C and 30.0 min. The oil yield is lower than that of commercial cultivars, but the contents of volatile and phenolic compounds is higher.     

Applicability of an In-Vitro Digestion Model to Assess the Bioaccessibility of Phenolic Compounds from Olive-Related Products

The Mediterranean diet includes virgin olive oil (VOO) as the main fat and olives as snacks. In addition to providing nutritional and organoleptic properties, VOO and the fruits (olives) contain an extensive number of bioactive compounds, mainly phenolic compounds, which are considered to be powerful antioxidants. Furthermore, olive byproducts, such as olive leaves, olive pomace, and olive mill wastewater, considered also as rich sources of phenolic compounds, are now valorized due to being mainly applied in the pharmaceutical and nutraceutical industries. The digestive system must physically and chemically break down these ingested olive-related products to release their phenolic compounds, which will be further metabolized to be used by the human organism. The first purpose of this review is to provide an overview of the current status of in-vitro static digestion models for olive-related products. In this sense, the in-vitro gastrointestinal digestion methods are widely used with the following aims: (i) to study how phenolic compounds are released from their matrices and to identify structural changes of phenolic compounds after the digestion of olive fruits and oils and (ii) to support the functional value of olive leaves and byproducts generated in the olive industry by assessing their health properties before and after the gastrointestinal process. The second purpose of this review is to survey and discuss all the results available to date.     

Determination of α-Tocopherol in Olive Oil by Solid-Phase Microextraction and Gas Chromatography–Mass Spectrometry

Olive oil has great human health benefits and is an important component of the Mediterranean diet. Its quality, sensory attributes, and oxidative stability are linked to the presence of minor compounds. Vitamin E (α-tocopherol) is a key component in these properties. In this work, solid-phase microextraction coupled to gas chromatography–mass spectrometry was used for the determination of α-tocopherol in olive oil. The analytical performance of the method has been assessed in fortified olive oil with negligible vitamin E concentrations. The calibration curve was linear from 0.020 to 0.500?mg/g. The limits of detection and quantification were 0.006 and 0.021?mg/g, respectively. Intraday and interday relative standard deviations were 3.2 and 10.0, respectively, and were concentration independent. The method was used for the determination of α-tocopherol in virgin and extra virgin olive oil, reporting average concentrations of 0.044?±?0.03 and 0.200?±?0.05?mg/g, respectively. Overall, the method is simple, sensitive, rapid, and solvent free, and provided high recoveries of 97.7?±?3.1%. In addition, vitamin E stability in extra-virgin olive oil was characterized by a shelf-life study.     

Chemical Diversity and Potential Target Network of Woody Peony Flower Essential Oil from Eleven Representative Cultivars (Paeonia × suffruticosa Andr.)

Woody peony (Paeonia × suffruticosa Andr.) has many cultivars with genetic variances. The flower essential oil is valued in cosmetics and fragrances. This study was to investigate the chemical diversity of essential oils of eleven representative cultivars and their potential target network. Hydro-distillation afforded yields of 0.11–0.25%. Essential oils were analyzed by GC-MS and GC-FID which identified 105 compounds. Three clusters emerged from multivariate analysis, representative of phloroglucinol trimethyl ether (‘Caihui’), citronellol (‘Jingyu’, ‘Zhaofen’ and ‘Baiyuan Zhenghui’) and mixed (the rest of the cultivars) chemotypes. ‘Zhaofen’ and ‘Jingyu’ also exhibited low levels of other rose-related compounds. The main components were subjected to a target network approach. Drug-likeness screening gave 20 compounds with predictive blood–brain barrier permeation. Compound target network identified six key compounds, namely nerol, citronellol, geraniol, geranic acid, cis-3-hexen-1-ol and 1-hexanol. Top enriched terms in GO, KEGG and DisGeNET were mostly related to the central nervous system (CNS). Protein—protein interactions revealed a core network of 14 targets, 11 of which were CNS-related (targets for antidepressants, analgesics, antipsychotics, anti-Alzheimer’s and anti-Parkinson’s agents). This work provides useful information on the production of woody peony essential oils with specific chemotypes and reveals their potential importance in aromatherapy for alternative treatment of CNS disorders.     

Comparative Evaluation of Different Targeted and Untargeted Analytical Approaches to Assess Greek Extra Virgin Olive Oil Quality and Authentication

Extra virgin olive oil (EVOO) is a key component of the Mediterranean diet, with several health benefits derived from its consumption. Moreover, due to its eminent market position, EVOO has been thoroughly studied over the last several years, aiming at its authentication, but also to reveal the chemical profile inherent to its beneficial properties. In the present work, a comparative study was conducted to assess Greek EVOOs’ quality and authentication utilizing different analytical approaches, both targeted and untargeted. 173 monovarietal EVOOs from three emblematic Greek cultivars (Koroneiki, Kolovi and Adramytiani), obtained during the harvesting years of 2018–2020, were analyzed and quantified as per their fatty acids methyl esters (FAMEs) composition via the official method (EEC) No 2568/91, as well as their bioactive content through liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) methodology. In addition to FAMEs analysis, EVOO samples were also analyzed via HRMS-untargeted metabolomics and optical spectroscopy techniques (visible absorption, fluorescence and Raman). The data retrieved from all applied techniques were analyzed with Machine Learning methods for the authentication of the EVOOs’ variety. The models’ predictive performance was calculated through test samples, while for further evaluation 30 commercially available EVOO samples were also examined in terms of variety. To the best of our knowledge, this is the first study where different techniques from the fields of standard analysis, spectrometry and optical spectroscopy are applied to the same EVOO samples, providing strong insight into EVOOs chemical profile and a comparative evaluation through the different platforms.     

Comparative study on fatty acid composition of olive (Olea europaea L.), with emphasis on phytosterol contents

The present study was designed to determine the fatty acid composition and phytosterol contents of Turkish native olive cultivars, namely Kilis Yağlık and Nizip Yağlık cv. In this context, olive fruits from 34 locations were sampled and then screened for their components in comparison. Fifteen different fatty acids were found in both olive oils. In the order of abundance, the most important ones were oleic acid (18:1) > palmitic acid (16:0) > linoleic acid (18:2) > stearic acid (18:0). Significant differences were observed in the contents of oleic acid (18:1), palmitic acid (16:0), linoleic acid (18:2) but not for stearic acid content in comparison both oils (p < 0.01). There were significant differences in terms of unsaturated fatty acids, saturated fatty acids and polyunsaturated fatty acids (p < 0.01). The seven phytosterols – cholesterol, campesterol, stigmasterol, β ‐sitosterol, Δ‐5‐avenasterol, Δ‐7‐stigmastenol and Δ‐7‐avenasterol – were studied in both oil sources. The predominant sterols were β ‐sitosterol, Δ5‐avenasterol and campesterol in the samples analysed. However, no significant differences were found in the levels of the phytosterols between the two olive cultivars.     

Synchronous fluorescence spectroscopy for quantitative determination of virgin olive oil adulteration with sunflower oil

Adulteration of extra virgin olive oil with sunflower oil is a major issue for the olive oil industry. In this paper, the potential of total synchronous fluorescence (TSyF) spectra to differentiate virgin olive oil from sunflower oil and synchronous fluorescence (SyF) spectra combined with multivariate analysis to assess the adulteration of virgin olive oil are demonstrated. TSyF spectra were acquired by varying the excitation wavelength in the region 270–720 nm and the wavelength interval (Δλ) in the region from 20 to 120 nm. TSyF contour plots for sunflower, in contrast to virgin olive oil, show a fluorescence region in the excitation wavelength range 325–385 nm. Fifteen different virgin olive oil samples were adulterated with sunflower oil at varying levels (0.5–95%) resulting in one hundred and thirty six mixtures. The partial least-squares regression model was used for quantification of the adulteration using wavelength intervals of 20 and 80 nm. This technique is useful for detection of sunflower oil in virgin olive oil at levels down to 3.4% (w/v) in just two and a half minutes using an 80-nm wavelength interval.     

Dried Destoned Virgin Olive Pomace: A Promising New By-Product from Pomace Extraction Process

At present the olive oil industry produces large amounts of secondary products once considered waste or by-products. In this paper, we present, for the first time, a new interesting olive by-product named “dried destoned virgin olive pomace” (DDVOP), produced by the pomace oil industry. The production of DDVOP is possible thanks to the use of a new system that differs from the traditional ones by having the dryer set at a lower temperature value, 350 °C instead of 550 °C, and by avoiding the solvent extraction phase. In order to evaluate if DDVOP may be suitable as a new innovative feeding integrator for animal feed, its chemical characteristics were investigated. Results demonstrated that DDVOP is a good source of raw protein and precious fiber; that it is consistent in total phenols (6156 mg/kg); rich in oleic (72.29%), linoleic (8.37%) acids and tocopherols (8.80 mg/kg). A feeding trial was, therefore, carried out on sheep with the scope of investigating the influence of the diet on the quality of milk obtained from sheep fed with DDVOP-enriched feed. The resulting milk was enriched in polyunsaturated (0.21%) and unsaturated (2.42%) fatty acids; and had increased levels of phenols (10.35 mg/kg) and tocopherols (1.03 mg/kg).     

Direct, reagent-free determination of free fatty acid content in olive oil and olives by Fourier transform Raman spectrometry

Fourier transform (FT) Raman spectrometry in combination with partial least squares (PLS) regression was used for direct, reagent-free determination of free fatty acid (FFA) content in olive oils and olives. Oils were directly investigated in a simple flow cell. Milled olives were measured in a dedicated sample cup, which was rotated eccentrically to the horizontal laser beam during spectrum acquisition in order to compensate sample heterogeneity. Both external and internal (leave-one-out) validation were used to assess the predictive ability of the PLS calibration models for FFA content (in terms of oleic acid) in oil and olives in the range 0.20-6.14 and 0.15-3.79%, respectively. The root mean square error of prediction (RMSEP) was 0.29% for oil and 0.28% for olives. The predicted FFA contents were used to classify oils and olives in different categories according to the European Union regulations. Ninety percent of the oil samples and 80% of the olives were correctly classified. These results demonstrate that the proposed procedures can be used for screening of good quality olives before processing, as well as, for the on-line control of the produced oil.     

Comparative Evaluation of the Phytochemical Profiles and Antioxidant Potentials of Olive Leaves from 32 Cultivars Grown in China

Olives (Olea europaea L.) are a significant part of the agroindustry in China. Olive leaves, the most abundant by-products of the olive and olive oil industry, contain bioactive compounds that are beneficial to human health. The purpose of this study was to evaluate the phytochemical profiles and antioxidant capacities of olive leaves from 32 cultivars grown in China. A total of 32 phytochemical compounds were identified using high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry, including 17 flavonoids, five iridoids, two hydroxycinnamic acids, six triterpenic acids, one simple phenol, and one coumarin. Specifically, olive leaves were found to be excellent sources of flavonoids (4.92–18.29 mg/g dw), iridoids (5.75–33.73 mg/g dw), and triterpenic acids (15.72–35.75 mg/g dw), and considerable variations in phytochemical content were detected among the different cultivars. All tested cultivars were classified into three categories according to their oil contents for further comparative phytochemicals assessment. Principal component analysis indicated that the investigated olive cultivars could be distinguished based upon their phytochemical profiles and antioxidant capacities. The olive leaves obtained from the low-oil-content (<16%) cultivars exhibited higher levels of glycosylated flavonoids and iridoids, while those obtained from high-oil-content (>20%) cultivars contained mainly triterpenic acids in their compositions. Correspondingly, the low-oil-content cultivars (OL3, Frantoio selection and OL14, Huaou 5) exhibited the highest ABTS antioxidant activities (758.01 ± 16.54 and 710.64 ± 14.58 mg TE/g dw, respectively), and OL9 (Olea europaea subsp. Cuspidata isolate Yunnan) and OL3 exhibited the highest ferric reducing/antioxidant power assay values (1228.29 ± 23.95 mg TE/g dw and 1099.99 ± 14.30 mg TE/g dw, respectively). The results from this study may be beneficial to the comprehensive evaluation and utilization of bioactive compounds in olive leaves.     

Rapid and quantitative extraction method for the determination of chlorophylls and carotenoids in olive oil by high-performance liquid chromatography

Colour is an organoleptic characteristic of virgin olive oil and an important attribute that affects the consumer perception of quality. Chlorophylls and carotenoids are the main pigments responsible for the colour of virgin olive oil. A simple analytical method for the quantitative determination of chlorophylls and carotenoids in virgin olive oils has been developed. The pigments were isolated from small samples of oil (1.0 g) by solid-phase extraction using diol-phase cartridges (diol-SPE), and the extract was analysed by reverse-phase HPLC with diode-array UV detection. Chromatographic peak resolution, reproducibility (coefficient of variation (C.V.) <4.5%) and recovery (>98.4%) for each component were satisfactory. A comparative study of the proposed method was performed versus classical liquid-liquid extraction (LLE) with N,N′-dimethylformamide and solid-phase extraction using a C18 column (C18-SPE). While 96.4% of the pigments were recovered by LLE, only 51.3% were isolated by C18-SPE in comparison to diol-SPE. Likewise, a higher alteration of pigment composition was observed when such LLE and C18-SPE procedures were used. In this sense, a higher ratio of pheophytin in comparison to that isolated by the diol-SPE procedure was achieved with both extraction procedures, indicating a greater extent of the pheophytinization reaction. Therefore, quantification of pigments from virgin olive oil by diol-SPE followed by RP-HPLC was found to be rapid, simple, required only a small amount of sample, consumed only small amounts of organic solvents, and provided high recoveries, accuracy and precision.     

Reuse of Food Waste: The Chemical Composition and Health Properties of Pomelo (Citrus maxima) Cultivar Essential Oils

The aim of the present study is to investigate the chemical profile, antioxidant activity, carbohydrate-hydrolysing enzyme inhibition, and hypolipidemic effect of essential oils (EOs) extracted from Sicilian Citrus maxima (pomelo) flavedo. Using gas-chromatography-mass spectrometry analysis (GC-MS) we analysed the Eos of five cultivars of C. maxima, namely, ‘Chadock’, ‘Maxima’, ‘Pyriformis’, ‘Terracciani’, and ‘Todarii’, and their blends. The antioxidant activity was performed by using a multi-target approach using 2,2′-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic acid (ABTS), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), ferric reducing ability power (FRAP), and β-carotene bleaching tests. The α-amylase, α-glucosidase, and lipase-inhibitory activities were also assessed. GC-MS analyses revealed D-limonene as the main monoterpene hydrocarbon in all cultivars, albeit with different percentages in the range of 21.72–71.13%. A good content of oxygenated monoterpenes was detected for all cultivars, especially for ‘Todarii’. The analysis of the principal components (PCA), and related clusters (HCA), was performed to find chemo-diversity among the analysed samples. EOs from ‘Chadock’ and ‘Maxima’ were statistically similar to each other, and they differed from P3 in the smaller amount of sesquiterpene hydrocarbons, while the oils from ‘Terracciani’ and ‘Todarii’ were found to be chemically and statistically different. ‘Chadock’ EO was the most active to scavenge radicals (IC₅₀ values of 22.24 and 27.23 µg/mL in ABTS and DPPH tests, respectively). ‘Terracciani’ EO was the most active against both lipase and α-amylase, whereas the blends obtained by the combination (1:1 v/v) of C. maxima ‘Maxima’ + ‘Todarii’ were the most active against α-glucosidase. Generally, the blends did not exert a unique behaviour in potentiating or reducing the bioactivity of the pomelo EOs.     

Extra Virgin Olive Oil Secoiridoids Modulate the Metabolic Activity of Dacarbazine Pre-Treated and Treatment-Naive Melanoma Cells

Nowadays, many individuals, whether healthy or diagnosed with disease, tend to expose themselves to various easily accessible natural products in hopes of benefiting their health and well-being. Mediterranean populations have traditionally used olive oil not only in nutrition but also in cosmetics, including skincare. In this study, the phenolic profile—composed of twelve compounds altogether, including the secoiridoids oleocanthal (OCAL) and oleacein (OCEIN)—of extra virgin olive oil (EVOO) from autochthonous cultivars from Croatia was determined using ¹H qNMR spectroscopy and HPLC-DAD analysis, and its biological activity was investigated in melanoma cell lines. The EVOO with the highest OCEIN content had the strongest anti-cancer activity in A375 melanoma cells and the least toxic effect on the non-cancerous keratocyte cell line (HaCaT). On the other hand, pure OCAL was shown to be more effective and safer than pure OCEIN. Post-treatment with any of the EVOO phenolic extracts (EVOO-PEs) enhanced the anti-cancer effect of the anti-cancerous drug dacarbazine (DTIC) applied in pre-treatment, while they did not compromise the viability of non-cancerous cells. The metastatic melanoma A375M cell line was almost unresponsive to the EVOO-PEs themselves, as well as to pure OCEIN and OCAL. Our results demonstrate that olive oils and/or their compounds may have a potentially beneficial effect on melanoma treatment. However, their usage can be detrimental or futile, especially in healthy cells, due to inadequately applied concentrations/combinations or the presence of resistant cells.     

Chemical Compositions and Antioxidant Activities of Essential Oils,and Their Combinations,Obtained from Flavedo By-Product of Seven Cultivars of Sicilian Citrus aurantium L.

In this work, seven Citrus aurantium essential oils (EOs) derived from flavedo of cultivars ‘Canaliculata’, ‘Consolei’, ‘Crispifolia’, ‘Fasciata’, ‘Foetifera’, ‘Listata’, and ‘Bizzaria’ were investigated. EOs were also combined in 1:1 (v/v) ratio to identify possible synergism or antagonism of actions. GC-MS analysis was done to investigate Eos’ phytochemical profiles. The antioxidant activity was studied by using a multi-target approach based on FRAP, DPPH, ABTS, and β-carotene bleaching tests. A great difference was observed in EOs’ phytochemical profiles. d-limonene (33.35–89.17%) was the main monoterpene hydrocarbon, and α-Pinene, β-myrcene, and β-linalool were identified in almost all samples. Among EOs, only C3 showed high quantitative and qualitative variability in its chemical composition. The chemical diversity of EOs was also demonstrated by PCA and HCA statistical analysis. Samples C2, C4, C5, C6, and C7 were statistically similar to each other, while C1 and C3 were characterized as having a different amount of other compounds and oxygenated monoterpenes, respectively, with respect to the other EOs mentioned. The global antioxidant score (GAS) revealed that among the tested EOs, C. aurantium ‘Fasciata’ EO had the highest antioxidant potential, with a GAS value of −0.47, whereas among combinations, the EO obtained by mixing ‘Canaliculata’ + ‘Bizzaria’ was the most active. Comparison by theoretical and real data on inhibitory concentration (IC₅₀) and FRAP values did not reveal any significant effect of synergism or antagonism of actions to be valid in all biological applied tests. These findings, considered together, represent an important starting point to understand which compounds are responsible for the activities and their future possible industrial application.