Bioactive compounds from Olea europaea: investigation and application in food, cosmetic and pharmaceutical industry

Periodic Reporting for period 2 - Olive-Net (Bioactive compounds from Olea europaea: investigation and application in food, cosmetic and pharmaceutical industry)

Periodo di rendicontazione: 2019-03-01 al 2022-08-31

The olive tree is a critical agricultural element for the EU, especially for the countries of the Mediterranean region. Olive oil and fruit contain hydrophilic (phenolic acids and alcohols, flavonoids and secoiridoids) and lipophilic compounds that are known to possess multiple biological activities. The main biophenols found in the products of olive tree are hydroxytyrosol, tyrosol, oleuropein, oleacein, oleocanthal, ligstoside aglycons and verbascoside. Furthermore, the olive processing industry is responsible for the production of a huge bulk of by-products. These materials, olive leaves and olive mill side products can also be proven as innovative and efficient sources of the aforementioned olive bioactives.
Olive-Net is an ambitious project based on state-of-the-art natural product chemistry and biology, in order to exploit diverse raw materials coming from the olive tree for the development of novel active ingredients that could be used in the pharmaceutical, nutraceutical and cosmetic industry.
This was achieved through an extended and well-balanced scheme of researcher’s secondments between universities and enterprises from EU & Associated countries as well as universities from Third countries. The objectives will be:
• Exploitation of diverse raw materials coming from the olive tree
• Generation of new extraction, enrichment and isolation protocols based on emerging novel technologies dedicated to olive bioactives
• Analyze the chemical content of the extracts/enriched fractions
• Investigation of the chemical transformation of olive bioactive compounds (such as oleocanthal and oleaceain) in order to improve their stability and pharmacological properties
• Investigation of the pharmacological properties of olive bioactives in relation to inflammation and ageing
• Perform PK/PD studies of pure compounds
• Development of new formulations of bioactive compounds and/or extracts for incorporation into pharmaceutical/nutraceutical/cosmeceutical products
• Scale up the best extraction process
• Address the demand of the consumers on natural solutions by formulating and testing new products based on plant chemical diversity.

The OliveNet project was implemented from 01/03/2017 to 31/08/2022, after also a period of suspension due to the Covid-19 pandemic. Specifically, the project was suspended on 23/07/2020 and on 02/2021 it was extended for 12 months, until 28/02/2022. Finally, the project was resumed on 01/06/2021 and an extra period of six months extension was given.
Seven work packages had been designed, involving experimental and administrative work. Thirteen partners participated as beneficiaries and three partners were from Third countries.
The project started with the acquisition of all relevant authorizations, approvals and certificates related to ethics requirement. In parallel, various samples of olive oil, fruits, leaves, pomace and olive mill waste waters (OMWW) were gathered from different countries (Greece, Algeria, Thailand) and were extracted with various techniques, such as liquid-liquid extraction, ultrasound extraction, maceration, accelerated solvent extraction and centrifugal partition extraction. Fractionation of the samples was accomplished with resins technology and centrifugal partition chromatography.
All the extracts were analyzed with TLC, HPLC-DAD and HPLC-MS analysis and were found to be rich sources of phenolic compounds, like hydroxytyrosol. The total phenolic content of the produced extracts and fractions was determined with Folin-Ciocalteu analysis while extracts, fractions and pure compounds were evaluated for their radical scavenging activity with DPPH assays. In continuation, the extracts/fractions/compounds were evaluated for their potential bioactivity. In vitro screening on the antioxidant potential and the anti-ageing effect was performed and again the extracts containing hydroxytyrosol where the most potent. The extract/fractions/compounds were also tested in cell based assays for their cellular antioxidant and anti-inflammatory potential and also for toxicity. The effects of oleocanthal and hydroxytyrosol were studied in selected animal models and further olive metabolites were determined in plasma samples using LC-HRMS approaches. Extracts with hydroxytyrosol were selected as the most promising and pilot scale extraction of these extracts was designed and implemented. A cosmeceutical product and a food supplement containing extracts with hydroxytyrosol were formulated and developed.
Dissemination activities implemented, involved the transfer of knowledge to all scientists working in the field of natural products and other stakeholders. Specifically, it involved the project website, the publication of articles in scientific journals, participation in scientific conferences, exhibitions and the organization of a workshop. The results of the project were disseminated also to the public through TV interviews.
Management activities involved all the activities and procedures required for the administration of the project, the communication between partners, the organization of meetings and the preparation and signature of the grant and partnership agreements. The kick off meeting was organized on 17/03/2017 and the mid-term meeting was held on the 30/11/18, in Athens with the presence of the project officer and an external expert. Several difficulties had been encountered regarding administrative issues related to the secondments. Mostly issues related to health insurance, visa, contracts etc had been shown to be really time-consuming and laborious. The first semester of 2020 was really difficult since most of the countries involved in the project were in a lock down situation with researchers working from home and couldn’t host people in their facilities. In addition, most of the airports were closed and flights were cancelled. The extension gave the opportunity to the consortium to organize and implement secondments at the last six months of the project, since most of the restrictions due to Covid-19 pandemic had arrested.

Progress beyond the state of the art
• Integration of advanced extraction techniques and enrichment technologies into the production of final products thus creating a new pipeline for the production of pharmaceuticals, nutraceuticals and cosmeceuticals
• Launch of a holistic approach for objectively assessing the efficacy of natural extracts, proving the scientific basis of their mode of action

Results
Olive-Net project generated promising entities that were integrated into novel cosmeceutical/nutraceutical/pharmaceutical products with well established properties

Impact
• Stimulate development of technologically innovative solutions, based on environmentally friendly technologies for exploiting olive-based material and olive polyphenols
• Improve the career prospects of researchers at all stages of their career by adding entrepreneurial skills to scientific expertise
• Boost collaborative networks within EU in an acute recession environment with the objective of exporting high added value products to third countries as well
• Increase awareness of the importance and benefit of new research approaches among policymakers and the general public in order to impact the decision making related to the support for innovative natural products development

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Periodic Reporting for period 2 - Olive-Net (Bioactive compounds from Olea europaea: investigation and application in food, cosmetic and pharmaceutical industry)

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