PROJECT IMPLEMENTATION PLAN

 1st Stage/2022

 

Year 2022 - Reporting period 01.07.2022 – 31.12.2022

 

Stage 1 - Development of experimental models for obtaining and characterizing selective bioactive fluid extracts from the fruits of bitter cherry species - Prunus avium var. sylvestris (L.) syn. Cerasus avium var. sylvestris (L.), blue honeysuckle - Lonicera caerulea (L.) and red grape pomace - Vitis vinifera (L.)

 

Verifiable estimated results of the stage:

- the web page of the project

- 3 experimental models developed of individual fluid extracts from bitter cherry fruits - Prunus avium var. sylvestris (L.) syn. Cerasus avium var. sylvestris (L.), blue honeysuckle - Lonicera caerulea (L.) and red grape pomace - Vitis vinifera (L.)

- 1 biotechnological model for the synergistic combination based on a mixture of fluid fruit extracts (MFE) with antimicrobial and antioxidant activity

- 1 scientific article in ISI journal (WoS)

- 1 communication at an international conference

 

Act 1.1 - Development of experimental fruit processing models, obtaining individual fluid extracts from bitter cherry fruits - Prunus avium var. sylvestris (L.) syn. Cerasus avium var. sylvestris (L.), blue honeysuckle - Lonicera caerulea (L.) and red grape pomace - Vitis vinifera (L.) and development of a biotechnological model for the mixture of fruit fluid extracts (MFE) obtained in optimal ratio.

 

Act 1.2 - Development of the experimental model for the physico-chemical characterization of individual fluid extracts from the fruits of blue honeysuckle – Lonicera caerulea (L.) and pomace from red grapes Vitis vinifera (L.), of the mixture of fluid extracts from fruits (MFE) and of barley germ powder.

 

Act 1.3 - Development and testing of experimental models for the evaluation of the biological activity of individual fluid fruit extracts and the mixture of fluid fruit extracts and the selection of optimal extracts with synergistic effect for encapsulation in mesoporous nanostructured matrices - part I.

 

Act 1.4 - Widely dissemination of results through communication at an international conference and publication of a scientific article in an ISI journal (WoS). Project web page.

 

STAGE no. 1 - Development of experimental models for obtaining and characterizing selective bioactive fluid extracts from the fruits of bitter cherry species - Prunus avium var. sylvestris (L.) syn. Cerasus avium var. sylvestris (L.), blue honeysuckle – Lonicera caerulea (L.) and red grape pomace – Vitis vinifera (L.)

 

 

STAGE EXECUTION PERIOD: 01 July – 31 December 2022

 

 

OBJECTIVES OF THE EXECUTION STAGE no. 1/2022

O1 – Development of experimental models and biotechnological variants for obtaining individual fluid extracts of bitter cherry, blue honeysuckle, pomace from fresh and preserved fruits and mixtures of fluid fruit extracts

O2 – Phytochemical and biological characterization and cytotoxicity evaluation of individual fruit fluid extracts of bitter cherry, blue honeysuckle, gooseberry, fruit fluid extract mixtures and barley germ prebiotic biomass

O3 – Dissemination of scientific results and project management

 

 

EXPECTED RESULTS

• 3 Optimized experimental models of individual fluid extracts from bitter cherry fruits - Prunus avium var. sylvestris (L.) syn. Cerasus avium var. sylvestris (L.), blue honeysuckle – Lonicera caerulea (L.) and red grape pomace – Vitis vinifera (L.)

• 1 Biotechnological model based on mixture of fluid fruit extracts (MFE) with antimicrobial and antioxidant activity suitable for nano-encapsulation in mesoporous silica matrix

• Active principles identified and quantified in individual fluid extracts of fruit, MFE and barley germ biomass prebiotics that confer the expected biological activity.

 

 

 

 

 

SUMMARY OF STAGE 1/2022

 

From the literature of the last decade, it has been observed that numerous reports have pointed out that the bioresources of pomace from red grapes, bitter cherry fruits and blue honeysuckle, as well as green barley germs are currently underexploited for biomedical purposes, as nutraceuticals. They present a significant content of bioactive principles with potential in hepato-digestive protection. The physico-chemical and biological evaluation of the extracts obtained from these bioresources, as such or in a mixture in different molar ratios, is of interest. The aim of this project stage was the selection of fluid extracts and mixtures of extracts with high bioactivity from the fruits of the bitter cherry species Prunus avium var. sylvestris (L.) syn. Cerasus avium (L.), blue honeysuckle Lonicera caerulea (L.) and the secondary product from winemaking –pomace from red grapes - Vitis vinifera (L.) by analyzing the content of bioactive principles and characterizing the antimicrobial, antioxidant activity and cytocompatibility in vitro with the help of sensitive and efficient experimental models. The main objectives of this stage of the project have been met, in accordance with the project implementation plan. Thus, experimental models of fluid extracts and a biotechnological model of a mixture based on fluid extracts from fruits (MFE), pomace and barley germs, selected for the high content of bioactive principles, with synergistic hepato-digestive protection action, were created . The individual extracts from bitter cherry and blue honeysuckle fruits, red grape pomace and barley germ were obtained in ethanol solvent of different concentrations, by means of the ultrasonication processing technique, with a reduced duration and energy consumption, but which ensures preservation better of fluid plant extracts. The biotechnological model consisted of mixtures of fluid extracts (MFE) of bitter cherry fruit, blue honeysuckle, Mamaia red grape pomace and barley germs, in ethanol solvent of different concentrations. The optimal individual fluid extracts and the mixture of fluid extracts were selected following the analysis of the active principles by determining the total content of phenolic compounds, in particular of total polyphenols, flavonoids, anthocyanins, tannins, total carotenoids, resveratrol, carbohydrates, proteins, vitamin C, trace elements, using sensitive UV-VIS, FTIR and AAS spectroscopy techniques. A comparative study was carried out to determine the antioxidant activity through complementary experimental models with different reaction mechanisms for the neutralization of two types of free radicals (ABTS, DPPH) for the mixture of fluid extracts and the individual extracts of the mixture components. It was shown that the samples contain antioxidant bioactive compounds, which could act synergistically to stop the propagation of the chain of oxidative reactions in the gastrointestinal tract. Following the in vitro tests of extract samples on bacterial and fungal cultures, their antimicrobial activity was observed. On a stabilized culture L929, the extract from the original variety Mamaia grape pomace was selected as the most biocompatible in the range of concentrations 10-500 µg/mL. Fluid extracts from bitter cherry and blue honeysuckle fruits, as well as from barley sprouts, were non-cytotoxic over the entire concentration range tested, respectively 10-1000 µg/mL. The active principles present in the mixture of extracts from the Mamaia variety of grape pomace, fruits and barley germs acted with a synergistic effect to improve cytocompatibility in L929 fibroblast culture. Through all these preliminary analyses, the biological potential of the mixture of fluid extracts and its usefulness in order to obtain new formulas of nanostructured nutraceuticals dispersed in a silica nanomatrix with a high content of functional bioactive agents of food quality (polyphenols, anthocyanins, carotenoids) have been demonstrated , flavonols, flavones, tannins, pectins, vitamins, trace elements) for hepato-digestive protection. The results obtained during this stage determined the achievement of all objectives, which are promising in the perspective of developing technologies for encapsulating the active principles from the prepared extracts, which have the expected therapeutic effects. In conclusion, within the first stage of the project, all the result indicators were reached, and the value of the results obtained is supported by the activity of disseminating the original results, by submitting to the publishing house one ISI (WoS) article, under evaluation; 3 papers communicated at international conferences, with abstracts published in Book of abstracts. A web page dedicated to the project was created at http://uniovidius.wixsite.com/nutrasinpro. The data obtained in the partnership within the project were centralized and integrated by the project coordinator within the technical and scientific report.