2G-lactic acid from olive oil supply chain waste: olive leaves upcycling via Lactobacillus casei fermentation.

The transition towards a sustainable model, particularly the circular economy, emphasizes the importance of redefining waste as a valuable resource, paving the way for innovative upcycling strategies. The olive oil industry, with its significant output of agricultural waste, offers a promising avenue for high-value biomass conversion into useful products through microbial processes. This study focuses on exploring new, high-value applications for olive leaves waste, utilizing a biotechnological approach with Lactobacillus casei for the production of second-generation lactic acid. Contrary to initial expectations, the inherent high polyphenol content and low fermentable glucose levels in olive leaves posed challenges for fermentation. Addressing this, an enzymatic hydrolysis step, following a preliminary extraction process, was implemented to increase glucose availability. Subsequent small-scale fermentation tests were conducted with and without nutrient supplements, identifying the medium that yielded the highest lactic acid production for scale-up. The scaled-up batch fermentation process achieved an enhanced conversion rate (83.58%) and specific productivity (0.26 g/L·h). This research confirms the feasibility of repurposing olive waste leaves for the production of lactic acid, contributing to the advancement of a greener economy through the valorization of agricultural waste. KEY POINTS: • Olive leaves slurry as it did not allow L. casei to ferment. • High concentrations of polyphenols inhibit fermentation of L. casei. • Enzymatic hydrolysis combined to organosolv extraction is the best pretreatment for lactic acid production starting from leaves and olive pruning waste.

Development and cross-validation of simple HPLC-fluorescence and UPLC-MS-UV methods for rapid determination of oleuropein in olive leaves.

INTRODUCTION: Olive leaves, abundant by-products of the olive oil industry, are a rich source of oleuropein, an important polyphenol in olive leaves. So far, no published methods have been validated using matrix standards for oleuropein quantification in olive leaves. OBJECTIVES: The study aimed to develop an HPLC method for oleuropein determination in olive leaves using spiked matrix standards prepared from a blank olive leaf matrix, to validate the method with respect to aqueous standards, and cross-validate the HPLC method with UPLC-MS and UPLC-UV techniques. METHODOLOGY: Oleuropein was extracted into methanol and analysed by HPLC with fluorescence detection (FLD; excitation and emission wavelengths 281 and 316 nm, respectively) and by UPLC-MS-UV. For validation, calibration curves of spiked matrix standards (0.4 to 4.8 mg/g) were analysed by the three methods over several days. Oleuropein was then analysed in French olive varieties. RESULTS: For the HPLC-FLD method, repeatability and intermediate precision were less than 5% RSD and linearity was demonstrated by the Fischer test. Differences in results of the spiked placebos by the three methods were non-significant, as confirmed by ANOVA. Extraction recovery was >90%, and there was a strong linear relationship between authentic and spiked matrix standards. The determination of oleuropein in French olive varieties is reported, including analysis in "Olivière" cultivar for the first time, leaves of which contained twice the amount of oleuropein compared with "Picholine". CONCLUSION: Accurate quantification of oleuropein is possible using aqueous standards. Cross-validation indicates that selective analysis can equally be carried out by HPLC or by UPLC-MS techniques.

Simultaneous High-Performance Recovery and Extended Acid-Catalyzed Hydrolysis of Oleuropein and Flavonoid Glycosides of Olive (Olea europaea) Leaves: Hydrothermal versus Ethanol Organosolv Treatment.

Olive leaves (OLLs) are an exceptional bioresource of natural polyphenols with proven antioxidant activity, yet the applicability of OLL extracts is constrained by the relatively high polarity of the major polyphenols, which occur as glycosides. To overcome this limitation, OLLs were subjected to both hydrothermal and ethanol organosolv treatments, fostered by acid catalysis to solicit in parallel increased polyphenol recovery and polyphenol modification into simpler, lower-polarity substances. After an initial screening of natural organic acids, oxalic acid (OxAc) was found to be the highest-performing catalyst. The extraction behavior using OxAc-catalyzed hydrothermal and ethanol organosolv treatments was appraised using kinetics, while treatment optimization was accomplished by deploying response-surface methodology. The comparative assessment of the composition extracts produced under optimal conditions of residence time and temperature was performed with liquid chromatography-tandem mass spectrometry and revealed that OLLs treated with 50% ethanol/1.5% HCl suffered extensive oleuropein and flavone glycoside hydrolysis, affording almost 23.4 mg hydroxytyrosol and 2 mg luteolin per g dry weight. On the other hand, hydrothermal treatment with 5% OxAc provided 20.2 and 0.12 mg of hydroxytyrosol and luteolin, respectively. Apigenin was in all cases a minor extract constituent. The study presented herein demonstrated for the first time the usefulness of using a natural, food-grade organic acid to perform such a task, yet further investigation is needed to maximize the desired effect.

The Bioactivity of a Hydroxytyrosol-Enriched Extract Originated after Direct Hydrolysis of Olive Leaves from Greek Cultivars.

Nowadays, olive leaf polyphenols have been at the center of scientific interest due to their beneficial effects on human health. The most abundant polyphenol in olive leaves is oleuropein. The biological properties of oleuropein are mainly due to the hydroxytyrosol moiety, a drastic catechol group, whose biological activity has been mentioned many times in the literature. Hence, in recent years, many nutritional supplements, food products, and cosmetics enriched in hydroxytyrosol have been developed and marketed, with unexpectedly positive results. However, the concentration levels of hydroxytyrosol in olive leaves are low, as it depends on several agricultural factors. In this study, a rapid and easy methodology for the production of hydroxytyrosol-enriched extracts from olive leaves was described. The proposed method is based on the direct acidic hydrolysis of olive leaves, where the extraction procedure and the hydrolysis of oleuropein are carried out in one step. Furthermore, we tested the in vitro bioactivity of this extract using cell-free and cell-based methods, evaluating its antioxidant and DNA-protective properties. Our results showed that the hydroxytyrosol-enriched extract produced after direct hydrolysis of olive leaves exerted significant in vitro antioxidant and geno-protective activity, and potentially these extracts could have various applications in the pharmaceutical, food, and cosmetic industries.

Evaluation of the effectiveness of natural extract as a substituent for synthetic preservatives and antioxidants in pharmaceutical preparations.

Background: Despite the fact that synthetic preservatives and antioxidants have strong antibacterial and antioxidant activity, they are frequently associated with negative health consequences. Currently, there is an increasing interest in pharmaceutical products that are excellent in quality and free of synthetic preservatives. Methods: As a result, the purpose of this research is to assess the antibacterial and antioxidant activities of olive leaf extract, oleuropein, and thymol in various pharmaceutical products. Furthermore, the efficacy of these natural extracts to substitute synthetic preservatives (methyl-propylparaben and benzalkonium chloride) and antioxidants (butylhydroxytoluene) will be investigated. Results: The results revealed that oleuropein, olive leaf extract, and a blend of oleuropein and thyme oil may be utilized as preservatives at concentrations of (0.6 % w/v), (0.4 % w/v), and (0.4 %/0.1 % v/v), respectively. The results demonstrated that thyme oil and oleuropein have synergistic efficacy against the studied microorganisms. By assessing antibacterial activity, and physical properties, the results demonstrated that pharmaceutical formulations containing natural preservatives were stable and effective for three months under accelerated settings (40 °C/75 % RH). Conclusion: Natural compounds such as oleuropein, olive leaf extract, and thyme oil have shown antibacterial effectiveness equivalent to synthetic preservatives in selected pharmaceutical products. Furthermore, there was synergy in antimicrobial activity between thyme oil and oleuropein and this facilitates the use of these compounds at different levels.

Preparation, characterization and wound-healing effect of PEGylated nanoliposomes loaded with oleuropein.

Chronic wounds have become a major concern for healthcare systems, as they have been related to diabetic foot ulcers, venous leg ulcers and pressure ulcers. Oleuropein is an active compound that is extracted from olive leaves and it has the ability to reduce injury to tissues owing to its antioxidant effect, hence improving wound healing. The poor pharmacokinetics of oleuropein have limited its use clinically. This work is aimed toward studying the impact of PEGylated and non-PEGylated nanoliposomes loaded with oleuropein, as a carrier model, on wound-healing activity. The thin film hydration method was used to compose PEGylated and non-PEGylated liposomes, both loaded with oleuropein. The results indicated that each free, PEGylated and non-PEGylated composition was within the limit of optimum nanoliposome characterization. The results showed that non-PEGylated compositions produced higher efficiency in encapsulation (47.09 ± 10.06%) than the PEGylated ones (20.97 ± 10.52%). The PEG-nanoliposomes loaded with oleuropein (PEG-oleu) had mean size, charge and polydispersity index of 129.35 nm, -9.55 mV and 0.1010, respectively. The scratch assay results proved that PEGylated liposomal compositions have a more rapid wound-healing activity than non-PEGylated ones at different time intervals at 0, 2, 24 and 28 h.

Molecular Mechanisms of the Protective Effects of Olive Leaf Polyphenols against Alzheimer's Disease.

Alzheimer's Disease (AD) is the cause of around 60-70% of global cases of dementia and approximately 50 million people have been reported to suffer this disease worldwide. The leaves of olive trees (Olea europaea) are the most abundant by-products of the olive grove industry. These by-products have been highlighted due to the wide variety of bioactive compounds such as oleuropein (OLE) and hydroxytyrosol (HT) with demonstrated medicinal properties to fight AD. In particular, the olive leaf (OL), OLE, and HT reduced not only amyloid-ß formation but also neurofibrillary tangles formation through amyloid protein precursor processing modulation. Although the isolated olive phytochemicals exerted lower cholinesterase inhibitory activity, OL demonstrated high inhibitory activity in the cholinergic tests evaluated. The mechanisms underlying these protective effects may be associated with decreased neuroinflammation and oxidative stress via NF-κB and Nrf2 modulation, respectively. Despite the limited research, evidence indicates that OL consumption promotes autophagy and restores loss of proteostasis, which was reflected in lower toxic protein aggregation in AD models. Therefore, olive phytochemicals may be a promising tool as an adjuvant in the treatment of AD.

Comparative Study of Novel Methods for Olive Leaf Phenolic Compound Extraction Using NADES as Solvents.

Natural deep eutectic solvents (NADES) composed of choline chloride with maltose (CMA), glycerol (CGL), citric (CCA) and lactic acid (CLA) combined with microwave (MAE), ultrasound (UAE), homogenate (HAE) and high hydrostatic pressure (HHPAE)-assisted extraction methods were applied to recover and compare olive leaf phenolic compounds. The resultant extracts were evaluated for their total phenol content (TPC), phenolic profile and antioxidant activity and compared with those of water and ethanol:water 70% v/v extracts. HAE was proven to be the most efficient method for the recovery of olive leaf phenolic compounds. The highest TPC (55.12 ± 1.08 mg GAE/g d.w.) was found in CCA extracts after HAE at 60 °C and 12,000 rpm, and the maximum antioxidant activity (3.32 ± 0.39 g d.w./g DPPH) was found in CGL extracts after UAE at 60 °C for 30 min. The TPCs of ethanol extracts were found to be higher than those of NADES extracts in most cases. The predominant phenolic compounds in the extracts were oleuropein, hydrohytyrosol and rutin.

In vitro and in vivo assessment of the anti-inflammatory activity of olive leaf extract in rats.

Inflammation is a complex and crucial process that protects the body against pathogens. Here in our study, we propose to scientifically justify the anti-inflammatory activity of olive leaf (OL). Initially, we ensured the safety of olive leaf extract (OLE) through acute oral administration of graded doses up to 4 g\kg in Wistar rats. Thus, the extract was considered generally safe. We also evaluated the ability of the extract to reduce carrageenan-induced rat paw edema. Compared to diclofenac sodium (10 mg/kg PO), OLE showed significant (P < 0.05) anti-inflammatory activity, showing the maximum inhibition percentage at the fifth hour of measurement at 42.31% and 46.99%, at doses of 200 and 400 m/kg, respectively, compared to 63.81% for the standard drug. To elucidate the potential mechanism, we measured TNF, IL-1, COX-2 and NO inside the paw tissue. Interestingly, OLE at all tested doses reduced the concentration of TNF and IL-1 to a level that was lower than that obtained by the standard drug. Additionally, OLE at the dose of 400 mg/kg reduced the levels of COX-2 and NO inside the paw tissue to a level that was statistically equivalent to the level observed in the normal control group. Finally, olive leaf extract at doses of 100, 200 and 400 mg/kg doses significantly (P < 0.05) inhibited the heat-induced hemolysis of RBCs membrane by 25.62, 57.40 and 73.88%, respectively, compared to 83.89% produced by aspirin. Consequently, we concluded that olive leaf extract has a significant anti-inflammatory activity through the reduction of TNF, IL-1, COX-2 and NO.

White rice enrichment with phenols upon cooking in olive leaf infusion: a preliminary study.

BACKGROUND: White rice is poor in health-promoting phytochemicals; therefore, the production of a phenol-enriched commodity is highly desirable. Recent findings on its enrichment via cooking in plant extracts are promising, yet studies employing aqueous extracts of olive leaves (OLs), containing well-recognized bioactive phenols (e.g. oleuropein) are absent. In addition, little is known about the levels of phenols that are maintained after rice drying and rehydration, an important aspect for the future design of 'ready-to-eat' functional rice. RESULTS: The examination, for the first time, of white rice adsorption capacity of phenols from OLs upon cooking in infusions containing different levels of phenols, after freeze-drying and rehydration, showed the following: (i) the total phenol content, the antioxidant activity (assessed via 2,2-diphenyl-1-picrylhydrazyl radical and ferric reducing antioxidant power assays), the oleuropein and luteolin-7-O-glucoside levels increased dose dependently; (ii) upon rehydration, the average decrease of total phenol content and antioxidant activity values was significantly lower when an exact volume of water was used compared with an excess (~10% versus 63%). A similar trend was observed for oleuropein (36% versus 83%) and the luteolin-7-O-glucoside (24 versus 82%) levels; (iii) the dried enriched kernels were less bright with a hay-yellow hue (CIELab coordinates). CONCLUSION: White rice enrichment with biophenols from OLs, a by-product of olive tree cultivation, was successful using a simple approach. Despite leaching upon freeze-drying/rehydration, sufficient amounts were maintained to obtain a functional rice that could serve as an alternative dietary source of OLs phenols to non-traditional olive tree product consumers or those refraining from sodium and fats. © 2023 Society of Chemical Industry.

Exploring the anthelmintic activity of Olea europaea L (Olive) leaves extract and oleuropein in mice naturally infected with Aspiculuris tetraptera.

Oxyuriasis, caused by the nematode Enterobius vermicularis, is one of the cosmopolitan intestinal infections of humans. Aspiculuris tetraptera commonly infects mice and it is morphologically similar to E. vermicularis. Parasitic resistance reduces the efficiency of synthetic drugs and poses economic impacts on the dairy sector, thus necessitating novel anthelmintic agents. Olea europaea L. (Olive) is a bioactive plant with potent pharmacological activities. However, its effects on oxyurids are poorly known, and no studies are currently exploring olives' anthelmintic potential. In this study, we investigated the pharmacokinetic behaviors of O. europaea leaves extract (OLE) and its phenolic compound oleuropein in mice infected with A. tetraptera, in comparison with Albendazole (ABZ), a standard drug used to treat parasitic worms. Fecal flotation method was used to identify the infestation with A. tetraptera eggs by examining the stool samples from mice. Infected animals were divided into 7 groups. 250 mg/kg, 500 mg/kg, and 1000 mg/kg doses of OLE, 5 mg/kg and 20 mg/kg doses of oleuropein, 10 mg/kg of ABZ and tap water were orally administered by gavage for 7 days during treatments. Drug efficacies and statistical differences between the treatments and controls were evaluated. Our results revealed 92.43 % efficacy of ABZ, similar to 92.19 % efficacy of 1000 mg/kg of OLE. At the same time, 250 mg/kg and 500 mg/kg concentrations of OLE remained 70.03 % and 63.18 % effective in reducing worm counts. Efficacy percentages of 5 mg/kg and 20 mg/kg of oleuropein were 9.27 % and 70.56 %, respectively. Statistical analysis of ABZ was significant compared to 1000 mg/kg of OLE, which was almost equal but insignificant. In general, our results confirm the anthelmintic potential of OLE and oleuropein against mice pinworms and open the way for targeted extraction of bioactive compounds from plants to optimize its use in human and veterinary medicine.

Comprehensive separation of a wide variety of compounds from olive leaves by counter-current chromatography with three-phase solvent system.

The three-phase solvent system counter-current chromatography has been of great research interest, because it can separate compounds with a wide range of polarity. The solvent system of n-hexane/methyl tert-butyl ether/acetonitrile/water (5:5:7:5, v/v) was used for counter-current chromatographic comprehensive separation of olive leaves. The study adopted the normal elution mode. The middle phase and the lower phase (at a volume ratio of 7:3) were pumped into the column simultaneously, followed by eluting with the upper, middle, and lower phases in sequence. The retention rate of the stationary phase measured by the experiment was 73.5%. The upper phase was used to elute the nonpolar compounds, then the mobile phase was switched to the middle phase to elute the moderately hydrophobic compounds, finally, the polar compounds were eluted by the lower phase remaining in the chromatographic column. This method successfully separated eight compounds in one step within 270 min and five compounds were identified. The logP values of these five compounds were 7.44, 7.86, 4.16, -0.11, and 0.96, respectively, covering a wide range of polarities. The present study demonstrated that the three-phase solvent has a strong extraction capacity for ingredients from extremely hydrophilic compounds to extremely hydrophobic compounds.

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.

Brazilian Agroindustrial Wastes as a Potential Resource of Bioative Compounds and Their Antimicrobial and Antioxidant Activities.

The study of the recovery of bioactive compounds from natural resources and its implications in several areas is very significant for the scientific community. This work aimed to study Brazilian agroindustrial wastes' antioxidant and antimicrobial activities using green extraction. Olive leaves, jaboticaba peel, araçá peel, and pecan nut shells were evaluated under four conditions: (1) convective-drying and aqueous extraction, (2) convective-drying and ethanolic extraction, (3) freeze-drying and aqueous extraction, and (4) freeze-drying and ethanolic extraction. The results demonstrated that all samples showed high antioxidant potential, and the highest antioxidant activity was obtained for the extract of pecan nut shell. As for the quantification of compounds by HPLC, the olive leaf presented the highest content of phenolic compounds in the extract, mainly oleuropein. Finally, the antimicrobial activity analysis revealed the extracts' bactericidal potential against Staphylococcus aureus and Escherichia coli. The present study shows that green extraction can extract bioactive compounds with antioxidant and antimicrobial properties, highlighting the importance of choosing the drying method and solvent for future uses of these natural resources by the industry.

Deriving valorization of phenolic compounds from olive oil by-products for food applications through microencapsulation approaches: a comprehensive review.

Nowadays, olive oil consumption is correlated to many health benefits, essentially due to the presence of antioxidants, especially phenolic compounds, which fostered its intensive production worldwide. During olive oil extraction, through continuous or discontinuous processes, many olive oil by-products are generated. These by-products constitute an environmental problem regarding its management and disposal. They are phytotoxic and biotoxic due to their high content of phenolic compounds, presenting contrastingly relevant health benefits due to their potent radical scavenging activities. In the framework of the disposal and management of olive oil by-products, treatment, and valorization approaches are found. As currently, the majority of the valorization techniques applied have a null market value, alternative strategies for the obtainment of innovative products as fortified foods are being investigated. The recovery and valorization strategies of olive oil by-products may comprise extraction and further encapsulation of bioactive compounds, as an innovative valorization blueprint of phenolic compounds present in these by-products. The majority of phenolic compounds present in olive oil by-products possess limited application on the food industry since they are promptly amended by environmental factors like temperature, pH, and light. Consequently, they must be protected previously ending in the final formulation. Prior to foods fortification with phenolic-rich extracts obtained from olive oil by-products, they should be protected through microencapsulation approaches, allowing a sustained release of phenolic compounds in the fortified foods, without losing their physicochemical properties. The combined strategies of extraction and microencapsulation will contribute to promoting the sustainability of the olive oil sector and aid the food industry to obtain reinvented added-value products.

Effect of olive leaves feeding on phenolic composition and lipolytic volatile profile in goat milk.

The aim of this study was to evaluate phenolic composition, antioxidant potential, and lipolytic events in raw milk obtained from goat fed a dietary supplementation with olive leaves (OL), a by-product of the olive oil production chain. For this purpose, 30 Saanen goats were randomly allocated into 2 groups of 15 goats each: the control group received a standard diet that was prepared by taking into account the nutritional needs of lactating goats, whereas the experimental group (EG) was fed with an OL-supplemented diet (10% on a dry matter basis). At the end of the 30 d of the trial, the individual milk samples were collected and immediately analyzed for total phenolic content and antioxidant activity (AOA). Subsequently, the individual phenolic compounds have been identified and quantified through an ultra-high-performance liquid chromatography system and a characterization of free fatty acids released in milk has been performed. The results showed a positive effect of dietary OL supplementation in improving total phenolic content and AOA; furthermore, 19 phenolic compounds, including phenolic acids, flavonoids, simple phenols, and secoiridoids, have been identified in EG milk. In addition to this, a reduced accumulation of free fatty acids has been found in EG milk, and this finding leads us to hypothesize an inhibitory action of the identified phenolic compounds toward the enzymes responsible for lipolytic events. The use of the molecular docking approach verified the interactions, defining a fairly interesting framework for cinnamic acid, which should be able to noncovalently bind these enzymes, interfering with the recruitment of the substrate and therefore, slowing down their hydrolytic activity. In any case, this information will be subjected to in vitro evaluations for an accurate characterization of the biochemical mechanisms that can be established in milk naturally enriched with bioactive compounds.

Quality assessment of leaf extracts of 12 olive cultivars and impact of seasonal variation based on UV spectroscopy and phytochemcial content using multivariate analyses.

INTRODUCTION: Recently, focus has been made on the health-oriented uses of olive leaves, a byproduct of olive production, as a potential source of antioxidants. Oleuropein is one of the phenolic components in olive leaves known for its high antioxidant value. OBJECTIVE: The main aim of the current study was constructing a model for the quality assessment of olive leaves and their potential phytochemical content and hence biological value as well. The phytochemical variation in olive leaves in both flowering (spring) and fruiting seasons (autumn) was also investigated. METHODS: In this study, the leaves of 12 different olive cultivars from different geographical origins growing in Egypt were assessed for their oleuropein content, total flavonoid (TF) content and total polyphenol (Pph) content in spring and autumn via ultraviolet (UV) spectroscopy and high-performance liquid chromatography (HPLC) coupled to multivariate data analyses. The antioxidant activity of olive leaf extracts was assessed using 2,2'-diphenyl-1-picrylhydrazyl (DPPH) assay. RESULTS: Higher levels of oleuropein, TF and Pph content were found in spring with the highest oleuropein content in the Spanish cultivar; Manzanillo, followed by the Italian cultivar Coratina and the Egyptian Agizi Okasi (218.94, 151.58 and 122.18 mg/100 g of dried leaf extract, respectively). UV spectra was also measured and the collected data were coupled to multivariate analyses showing clustering of cultivars with common geographical origin. CONCLUSION: Our findings emphasised the influence of collection time and type of cultivar on the chemical profile of olive leaves. The model presented herein, serves for the quality assessment of olive leaves based on their phytochemical profile.

HPTLC and FTIR Fingerprinting of Olive Leaves Extracts and ATR-FTIR Characterisation of Major Flavonoids and Polyphenolics.

The aim of this study was to analyse the effect of spontaneous microbial maceration on the release and extraction of the flavonoids and phenolics from olive leaves. Bioprofiling based on thin-layer chromatography effect-directed detection followed by ATR-FTIR spectroscopy proved to be a reliable and convenient method for simultaneous comparison of the extracts. Results show that fermentation significantly enhances the extraction of phenolic compounds and flavonoids. The polyphenolic content was increased from 6.7 µg GAE (gallic acid equivalents) to 25.5 µg GAE, antioxidants from 10.3 µg GAE to 25.3 µg GAE, and flavonoid content from 42 µg RE (rutin equivalents) to 238 µg RE per 20 µL of extract. Increased antioxidant activity of fermented ethyl acetate extracts was attributed to the higher concentration of extracted flavonoids and phenolic terpenoids, while increased antioxidant activity in fermented ethanol extract was due to increased extraction of flavonoids as extraction of phenolic compounds was not improved. Lactic acid that is released during fermentation and glycine present in the olive leaves form a natural deep eutectic solvent (NADES) with significantly increased solubility for flavonoids.

Olive-Derived Triterpenes Suppress SARS COV-2 Main Protease: A Promising Scaffold for Future Therapeutics.

SARS CoV-2 pandemic is still considered a global health disaster, and newly emerged variants keep growing. A number of promising vaccines have been recently developed as a protective measure; however, cost-effective treatments are also of great importance to support this critical situation. Previously, betulinic acid has shown promising antiviral activity against SARS CoV via targeting its main protease. Herein, we investigated the inhibitory potential of this compound together with three other triterpene congeners (i.e., ursolic acid, maslinic acid, and betulin) derived from olive leaves against the viral main protease (Mpro) of the currently widespread SARS CoV-2. Interestingly, betulinic, ursolic, and maslinic acids showed significant inhibitory activity (IC50 = 3.22-14.55 µM), while betulin was far less active (IC50 = 89.67 µM). A comprehensive in-silico analysis (i.e., ensemble docking, molecular dynamic simulation, and binding-free energy calculation) was then performed to describe the binding mode of these compounds with the enzyme catalytic active site and determine the main essential structural features required for their inhibitory activity. Results presented in this communication indicated that this class of compounds could be considered as a promising lead scaffold for developing cost-effective anti-SARS CoV-2 therapeutics.

Optimization of ultrasound-assisted aqueous two phase extraction of polyphenols from olive leaves.

In this study, polyphenols from olive leaves was extracted by ultrasound-assisted aqueous two phase extraction (UAATPE). Based on single factor experiment and response surface methodology (RSM), the optimum extraction conditions of polyphenols contained 29% (w/w) (NH4)2SO4, 35% (w/w) ethanol, pH 6.7, and 45 °C. The maximum extraction yield of polyphenols and oleuropein content were 34.06 mg/g and 44.13 mg/L, respectively. Compared with ultrasound-assisted extraction (UAE) and aqueous two phase extraction (ATPE), the extraction yield of polyphenols by UAATPE was 9.48 and 61.19% higher, respectively. In addition, the extract of UAATPE had higher purity. The results of antioxidant activity showed that polyphenols extracted by UAATPE had stronger DPPH and hydroxyl radicals scavenging ability and reducing power. Therefore, UAATPE is an efficient method for extracting polyphenols from olive leaves.