PN-III-P2-2.1-PED-2019-3037
Funded by a grant of the Romanian Ministry of
Education and Research, CCCDI - UEFISCDI, project number PN-III-P2-
2.1-PED-2019-3037, within PNCDI III, contract number 385PED
PROJECT CODE: PN-III-P2-2.1-PED-2019-3037, contract number 385PED ⁄ 2020
PROJECT TITLE:
Intelligent and active systems in food packaging based on biopolymers and novel flavylium dyes
Sisteme inteligente si active pentru impachetarea alimentelor, bazate pe biopolimeri si coloranti noi de tip ioni de flaviliu
ACRONYM: SMARTBIOPACK
PROJECT COORDINATOR: Politehnica University of Timişoara
PROJECT DIRECTOR: Conf. Dr. Ing. Mihai Medeleanu
BUDGET: 595.000 RON (Public budget)
START TIME: 01/11/2020
END TIME: 31/10/2022
PROJECT LAYOUT
The food industry faces two major problems: the accumulation of synthetic plastic and food waste, so the research interest for intelligent and active systems, as well as biodegradable materials has constantly increased in recent years. Cost efficient biodegradable and edible packaging may be the friendly solution to the environment, while active and intelligent packaging providing valuable information on the quality and safety of the food products appear to be the solution to reduce food waste and increase the protection of the consumers.
The main scope of the project is to develop a demonstration model for production of new materials used in food packaging systems based on biopolymers and synthetic dyes from flavylium derivatives as indicators and monitor of packed food quality. To obtain these materials, the synthesis and evaluation of photochromic properties of some novel dye derivatives of the flavylium ion based on an initial procedure of the properties evaluation by computational methods will be carried on. The synthetic dyes after full physicochemical characterization and photochromic properties evaluation with pH and thermal variation, will be mixed with biopolymers (based on chitosan or a blend of chitosan with polyvinyl alcohol or polylactic acid) to obtain the needed packaging material which will be tested again to determine the behavior in contact with food.
PROJECT TEAM:
Project leader: Conf. Dr. Ing. Mihai Medeleanu
Reasearchers:
Ș.l. dr. ing. Iulia Maria Păușescu
Ș.l. dr. ing. Anamaria Todea
Ș.l. dr. ing. Valentin Badea
PhD Students:
Drd. Eng. Ionut-Mihai Tănase
Drd. Eng. Ioan Bîtcan
OBJECTIVE: The main objective of the project is to develop a demonstration model for production of new materials used in food packaging systems based on biopolymers and synthetic dyes from flavylium derivatives as indicators and monitor of packed food quality.
STAGES
Stage 1 (2020, 2 months) – Design of flavylium dyes
Stage 2 (2021, 12 months) – Synthesis and characterization of proposed flavylium dyes and preparation and characterization of films based on biopolymers and the synthesized dyes
Stage 3 (2022, 10 months) – Evaluation of the properties of the films based on biopolymers and the synthesized dyes for food packaging applications.
DELIVERABLES:
Stage 1
A list of proposed compounds with all the computed properties, ordered by the two type of activities: antioxidant and photochromic;
Stage 1 report.
Stage 2
Synthesis protocol for the novel flavylium based dyes;
Protocol for the pH-dependent photochromic properties;
Protocol for the antioxidant properties of the synthesized dyes;
A set of flavylium based dyes with full physicochemical characterization and antioxidant, photochromic and toxicological properties evaluated – grams quantities;
Scientific article submitted to an ISI quoted journal;
Abstract for one presentation at scientific conference in the field.
Stage 2 report.
Stage 3
Gram quantities of obtained films together with their characterization by depicted methods;
Abstracts for two presentations at scientific conferences in the field;
Scientific article submitted to an ISI quoted journal
Stage 3 report
Final report.
RESULTS
Stage 1 (2020, 2 months) – Design of flavylium dyes
Abstract:
The objectives:
A list of proposed compounds with all the computed properties, ordered by the two type of activities: antioxidant and photochromic; Stage 1 report.
In this stage two series of flavylium type compounds were selected: S1 based on the structure of malvidine and S2 based on peonidine. For these compounds some computational studies were made in order to evaluate their specific properties like stability, reactivity, antioxidant activity, photochromic properties and also evaluation of their possible toxicity. The computations were done by Gaussian09 package on 6-31+G(d,p) basis set and the DFT-B3LYP method and the results were a list of molecular descriptors for chemical reactivity: HOMO-LUMO gap, electronegativity, absolute hardness and electrophilicity. For the S1 series compounds the photochromic properties were evaluated by prediction of UV-VIS spectrum for the main chemical species that arise during the spectral radiation. The predictions were made also by Gaussian09 with 6-31+G(d,p) in the frame of TD-DFT (Time dependent DFT) and water as solvent. The antioxidant properties were evaluated by computing some energetic molecular descriptors like BDE- bond dissociation enthalpy of O-H bond, IP-ionization potential, PDE-Proton dissociation enthalpy in O-H system, PA-proton affinity, ETE-electron transfer enthalpy, all associated with three reactions mechanisms that characterize the antioxidant reactivity of phenolic structures: HAT- hydrogen atom transfer, SET-PT single-electron transfer – proton transfer, SPLET - sequential proton loss electron transfer. The possible toxicity of S1 compounds were estimated by OSIRIS Property Explorer, a system which evaluate the toxicity risks like mutagenic, tumorigenic, irritant and reproductive effective together with some drug-relevant properties (octanol/water partition coefficient, solubility, molecular weight, topological polar surface area, drug-likeness and drug-score (bioactivity score)). The results of these computations were analyzed and four compounds from S1 series were proposed for synthesis.
All the objectives of this stage were fulfilled.
Stage 2 (2021, 12 months) – Synthesis and characterization of proposed flavylium dyes and preparation and characterization of films based on biopolymers and the synthesized dyes
Abstract:
The objectives: Synthesis protocol for the novel flavylium based dyes; Protocol for the pH-dependent photochromic properties; Protocol for the antioxidant properties of the synthesized dyes; A set of flavylium based dyes with full physicochemical characterization and antioxidant, photochromic and toxicological properties evaluated – grams quantities; Scientific article submitted to an ISI quoted journal; Abstract for one presentation at scientific conference in the field. Stage 2 report.
In this stage the computational studies were finalized for the prediction of antioxidant activity, photochromic properties and also evaluation of the possible toxicity of the S2 series of flavylium dyes type. The procedure was similar to the studies in the stage 1.
The computations were done by Gaussian09 package on 6-31+G(d,p) basis set and the DFT-B3LYP method and the results were a list of molecular descriptors for chemical reactivity: HOMO-LUMO gap, electronegativity, absolute hardness and electrophilicity. For the S2 series compounds the photochromic properties were evaluated by prediction of UV-VIS spectrum for the main chemical species that arise during the spectral radiation. The predictions were made also by Gaussian09 with 6-31+G(d,p) in the frame of TD-DFT (Time dependent DFT) and water as solvent. The antioxidant properties were evaluated by computing some energetic molecular descriptors like BDE- bond dissociation enthalpy of O-H bond, IP-ionization potential, PDE-Proton dissociation enthalpy in O-H system, PA-proton affinity, ETE-electron transfer enthalpy, all associated with three reactions mechanisms that characterize the antioxidant reactivity of phenolic structures: HAT- hydrogen atom transfer, SET-PT single-electron transfer – proton transfer, SPLET - sequential proton loss electron transfer. The possible toxicity of S1 compounds were estimated by OSIRIS Property Explorer, a system which evaluate the toxicity risks like mutagenic, tumorigenic, irritant and reproductive effective together with some drug-relevant properties (octanol/water partition coefficient, solubility, molecular weight, topological polar surface area, drug-likeness and drug-score (bioactivity score)). The results of these computations were analyzed and four compounds from S2 series were proposed for synthesis.
Based on the previous computations studies 8 selected compounds were synthesized (4 form S1 series and 4 from S2 series). They were analyzed and characterized by spectroscopic methods: FT-IR, UV-VIS, 1H-NMR, 13C-NMR, 2D-NMR and by thermogravimetric methods. By UV-VIS spectroscopy the dependence of photochromic properties under pH variation in time were evaluated. For the observed transformations a set of reversible reactions were proposed and the halochromic effect of these compounds was emphasized. For antioxidant properties the colorimetric method with DPPH was used. For two of the 8 compounds (S1_4 and S2_4) the results were even better than those obtained for some conventional standard antioxidants (vitamin C and curcumin). For 6 of the compounds cytotoxic properties were investigated on cell lines containing tumor cells type HT29, specific for colon cancer. Two compounds (S1_3 and S2_3) provided a higher toxicity against the tumor cell. However the others had no significant activity.
Insertion of dyes in polymer film implies two kind of films: monocomponent (based on chitosan with 4 of the synthesized dyes: S1_4, S2_2, S2_3, S2_4) and polycomponent (based by mixtures chitosan-polyvinyl alcohol, chitosan-cellulose and chitosan starch using S1_4, S1_10, S2_2 and S2_3 dyes). The films were characterized by FT-IR and UV-VIS spectroscopy and by thermogravimetric methods for thermal properties. For obtained polymeric films three material properties were measured: water content, solubility in water and swollen degree.
The results were presented in 3 international conferences (two abroad and one in Romania) and one paper sent to an ISI journal (under evaluation).
All the objectives of this stage were fulfilled.
Stage 3 (2022, 12 months) – Evaluation of the properties of the films based on biopolymers and the synthesized dyes for food packaging applications.
Abstract
In this stage the antioxidant properties of the synthesized dyes has been done (dyes S1_2, S1_3, S1_10, S2_2, S2_3 and S2_10) using the colorimetric method with DPPH. The IC50 parameter was computed. This parameter is defined as the antioxidant concentration value that produce a decrease with 50% of the DPPH solution absorbance. The flavylium salts from the Series 1 were much more reactive comparing with those from Series 2. Comparing with curcumin antioxidant activity, all the compounds from Series 1 had higher activity, and comparing with ascorbic acid activity (C vitamin) all but S1_10 compound had higher activity (values around the ascorbic acid activity).
For some of the synthesized compounds (S2_2, S2_3, S2_4, S2_1) cytotoxic properties were investigated on cell series containing cancerous cells HepG2 and HCT116, specific for liver cancer (HepG2) and colorectal cancer (HCT116) respectively, very relevant in the studies on intestinal microbiota. At concentration 0.132 mM almost all the compounds effective inhibit the growth of cancerous cells, except the S2_4 compound in HepG2 cell series, for which the percent of viable cells was 95.5%.
Monocomponent polymer film were synthesized based on chitosan, containing four of the previous synthesized dyes S1_2, S1_3, S1_10, S2_10. These were characterized by FT-IR and UV-VIS spectroscopy, and the thermal properties were studied by thermogravimetric analysis.
Two-component polymer films were synthesized based on chitosan-PVA, chitosan-cellulose and chitosan-starch mixtures, containing S1_2, S1_3, S2_10, S2_4 dyes. These were also characterized by FT-IR and UV-VIS spectroscopy, and the thermal properties by thermogravimetric analysis.
For the obtained polymer films, three material properties were evaluated: the water content, the solubility in water and swelling degree.
To evaluate the color response of the polymer films to pH change in water solutions, small probes (1x1 cm) from those films were immersed in 10 mL buffer solutions.The pH values were 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 and UV-VIS spectra were recorded for buffer solutions alone and for films. The dyes desorption was evaluated after 1h, 2h and 24h after immersion.
The sensitivity of the films in monitoring the food quality wad tested in two cases: direct contact with food matrix and contact with the package environment atmosphere on two food matrices: refrigerated chicken meat and swine meat.
The results of this stage were presented at two international conferences and three articles in scientific journals (Q1 quartile and total impact factor 14.93)
All the objectives of this stage were fulfilled.
Conferences:
- Corneanu C., Păușescu I., Todea A., Peter F., Medeleanu M., Ledeți I., Chitosan/PVA pH indicator films with potential food packaging applications, Advances in Food Chemistry Conference, AdFoodChem, 15-17 Aprilie 2021, online.
- Corneanu C., Păușescu I., Peter F., Medeleanu M., Synthesis, characterization and halochromism of a new xanthylium derivative, International Conference on Photochemistry – Ediția a 30-a, IPC30, 19-23 Iulie 2021, online.
- Bîtcan I., Argetoianu R., Dăescu D., Dreavă D.-M., Medeleanu M., Păușescu I., Bio-based pH indicator films for intelligent food packaging applications, EuroFoodChem – Ediția a 21-a, 22-24 Noiembrie 2021, online.
- Păușescu I., Dăescu D., Dreavă D.-M., Bîtcan I., Medeleanu M., Theoretical and experimental studies on the antioxidant properties of new flavylium dyes, 33rd International Course and Conference on the Interfaces among Mathematics, Chemistry and Computer Sciences: Mathematics, Chemistry, Computing – Math/Chem/Comp, MC2-33, 6-10 June 2022, Dubrovnik. Croația.
- Păușescu I., Dreavă D.-M., Bîtcan I., Dăescu D., Medeleanu M., pH indicator films based on renewable biodegradable polymers for food packaging applications, 9th IUPAC International Conference on Green Chemistry – 9th ICGC, 4-9 September 2022, Atena, Grecia.
Publications:
- Păușescu I., Kántor I., Babos G., May Z., Fodor-Kardos A., Miskolczy Z., Biczók L., Péter F., Medeleanu M., Feczkó T., Halochromic Behavior and Anticancer Effect of New Synthetic Anthocyanidins Complexed with -Cyclodextrin Derivatives, International Journal of Molecular Sciences, 2022, 23, 8103. https://doi.org/10.3390/ijms23158103.
- Păușescu I., Dreavă D.-M., Bîtcan I., Argetoianu R., Dăescu D., Medeleanu M., Bio-based pH indicator films for intelligent food packaging applications, Polymers, 2022, 14, 3622. https://doi.org/10.3390/polym14173622.
- Păușescu I., Todea A., Dreavă D., Boboescu T., Pațcan B., Pațcan L., Albulescu D., Badea V., Peter F., Tőtős R., Ursu D., Szolga L., Medeleanu M., Experimental and computational studies on bio-inspired fla-vylium salts as sensitizers for dye-sensitized solar cells, Materials, 2022, 15, 6985. https://doi.org/10.3390/ma15196985
Prezentarea succintă a rezultatelor obținute în cadrul proiectului
Proiectul cu titlul „Sisteme inteligente și active pentru împachetarea alimentelor, bazate pe biopolimeri și coloranți noi de tip ioni de flaviliu” a avut ca scop dezvoltarea unui model demonstrativ pentru producția de materiale noi utilizate la împachetarea alimentelor, care să conțină compuși ce pot monitoriza și oferi informații asupra posibilelor transformări ale alimentelor împachetate. Validarea modelului s-a făcut prin testarea materialelor în contact cu produsele alimentare împachetate conform descrierii unor reglementări ale Uniunii Europene.
Obiectivul principal a fost obținerea materialelor noi pentru împachetarea alimentelor, materiale care au la bază biopolimeri care conțin coloranți sintetici derivați ai ionului de flaviliu. Pornind de la faptul că ionul de flaviliu este structura de bază a antocianidinelor (compuși naturali cu puternice efecte antioxidante) au fost sintetizați coloranți pe bază de ioni de flaviliu pentru a fi amestecați cu biopolimeri pentru obținerea filmelor care să monitorizeze calitatea alimentelor împachetate datorită proprietăților fotocromice și antioxidante ale coloranților. Proprietățile fotocromice permit monitorizarea produselor alimentare datorită modificărilor de culoare ce apar ca urmare a modificării acidității mediului (pH-ul) în urma unor posibile transformări ale alimentelor.
Materialele obținute, prin conținutul de colorant cu proprietăți fotocromice s-au dovedit sensibile la variațiile de pH în timp, prin modificarea culorii datorate degradărilor posibile ale alimentelor. Derivații de flaviliu au fost sintetizați după o procedură inițială de evaluare a proprietăților fotocromice și antioxidante prin metode computaționale. După selecția compușilor s-a efectuat sinteza lor, caracterizarea fizico-chimică (spectroscopie UV-VIZ, FT-IR, RMN) pentru demonstrarea structurii lor și evaluarea proprietăților fotocromice și antioxidante. Coloranții obținuți au fost încorporați în biopolimerii de tip chitosan, alcool polivinilic, celuloză și amestecuri ale acestora. Filmele obținute au fost caracterizate prin procedee fizico-chimice (spectroscopie și analiză termică). A fost testată capacitatea de monitorizare a modificărilor de culoare inițial doar prin urmărirea modificărilor aplicate mediului în care se află filmul polimeric, apoi monitorizarea în contact cu alimentele. Produsele rezultate au fost testate și în privința proprietăților toxicologice.
S-a demonstrat utilitatea acestor combinații de filme bazate pe biopolimeri și coloranți cu proprietăți fotocromice și antioxidante la monitorizarea evoluției calității alimentelor în timp, fapt care permite dezvoltarea în continuare a cercetărilor în direcția obținerii unor materiale noi.
Figura 1. Filme chitosan-alcool polivinilic-colorant de tip ion de flaviliu
la diferite valori ale pH-ului pentru demonstrarea efectului fotocromic
Figura 2. Modul de realizare a probelor pentru determinarea sensibilității filmelor la modificarea calității
produselor alimentare (filme în contact direct cu alimentele sau în contact cu atmosfera de ambalare)