Edible biofilm containing red propolis extract for cosmetic use and for coating and preserving fruits and vegetables.

Abstract

"Edible biofilm enriched with red propolis: obtaining process, composition, and application in the preservation of fruits and vegetables" proposes edible biofilms enriched with red propolis extract (bcpv), a process for obtaining bcpv, and a process for applying bcpv to fruits and vegetables. The production of bcpv belongs to the field of food technology, more specifically to the area of ​​food packaging and food safety. Edible biofilms enriched with hydroalcoholic extract of red propolis (bcpv), a process for obtaining bcpv, and a process for applying bcpv to coat fruits and vegetables are proposed. The production of bcpv is linked to the use of standardized hydroalcoholic extract of red propolis, followed by the technique for obtaining film-forming solutions and their use as a coating for fruits and vegetables.BCPVs (Bottom-Coated Polypropylene) have applications in the food industry, mainly due to their ability to reduce gas exchange, reduce respiration rate, reduce ripening speed, in addition to their antimicrobial and antioxidant properties. Their use also generates cost reductions and reduces waste generation associated with primary packaging.

Description

1 / 29 Descriptive Report “Edible Biofilm Containing Red Propolis Extract for Cosmetic Use and for Coating and Preserving Fruits and Vegetables”

[001] The present invention belongs to the fields of Food Technology and Pharmaceuticals, more specifically to the area of ​​natural products with applicability in the preservation of fruits and vegetables. An edible biofilm composed of hydroalcoholic extract of red propolis (BCPV), a process for obtaining BCPV, pharmaceutical compositions containing BCPV, a process for obtaining pharmaceutical compositions containing BCPV, and uses are proposed. The production of BCPV is linked to the use of a standardized hydroalcoholic extract of red propolis, followed by the technique for obtaining biofilms, presented in the form of an edible biofilm. Additionally, the present application deals with pharmaceutical compositions containing said BCPV.BCPVs (Biologically Capillary-Vitamin Enamel) have applications in the food and pharmaceutical industries, mainly due to their ability to slow down ripening and their antioxidant and antimicrobial activity, especially as a coating for fruits and vegetables. FUNDAMENTALS OF THE INVENTION Acerola

[002] Products such as fruits and vegetables typically exhibit high perishability. Factors such as climatic patterns and peel fragility accelerate the senescence and deterioration process of these products, which are more susceptible to damage caused by mechanical impacts and injuries, increasing the risk of deterioration and reducing their post-harvest shelf life (OLIVEIRA et al. Post-harvest conservation of carambola under refrigeration with gelatin and PVC biofilm coating. 2015; ATAÍDE et al. Carnauba wax and propolis in the post-harvest conservation of juazeiro fruits under refrigerated conditions. 2017). Petition 870260037234, dated 04 / 22 / 2026, page 12 / 41 2 / 29

[003] Acerola (Malpighia emarginata), also known as West Indian cherry, is a fruit native to South and Central America. It was introduced to northeastern Brazil about 60 years ago by the Federal University of Pernambuco, through seeds brought from Puerto Rico (SILVA, MFV Effect of different treatments and packaging on the characteristics of acerola pulp and on the determination of ascorbic acid and anthocyanin content during storage. 1999. 245 p. Thesis (Doctorate in Food Technology) - Faculty of Food Engineering, State University of Campinas, Campinas, 1999).

[004] Its cultivation has increased considerably in the Northeast region, mainly due to the good soil and climate conditions.

[005] Planting conditions in the Northeast allow for fruit cultivation almost year-round, which is why it is the region with the highest acerola production in Brazil (about 64% of national production), and Brazil is the world's largest producer and exporter (EMBRAPA. A Acerola Crop. Brasília: Embrapa, 2012. 150 p).

[006] Acerola trees have 3 to 4 harvests per year. The fruits weigh about 4 g and their color varies from light green to bright dark red, depending on the degree of ripeness (Figure 1). Approximately 80% of the total weight of the fruit is edible (FURLANETO, FPB; NASSER, MD Panorama da cultura da acerola no estado de São Paulo. Pesquisa & Tecnologia, v. 12, n. 1, p. 1-6, 2015).

[007] Acerola is rich in vitamin C, containing approximately 50 mg / g of pulp (EMBRAPA. A Cultura da Acerola. Brasília: Embrapa, 2012. 150 p), surpassing other fruits considered excellent sources of ascorbic acid, such as oranges, cashews, and guavas. Therefore, consuming 2 to 3 fresh acerola fruits provides the minimum recommended daily dose of vitamin C, which is approximately 100 mg (amount varies depending on the country and other factors). Petition 870260037234, dated 04 / 22 / 2026, page 13 / 41 3 / 29 factors, such as age, sex, nutritional status, lactation, etc.) (AZULAY, MM et al. Vitamin C. Continuing Medical Education, v. 78, n. 3, p. 265-274, 2003).

[008] Vitamin C, or ascorbic acid, is one of the most significant nutritional components in fruits, although its content generally does not exceed 0.4%, with some exceptions, such as acerola (1.0 to 1.8%), and camu-camu (2.5 to 6.0%), which has the highest content among fruits (OLIVEIRA, LA Vitamin C, In: Laboratory Manual: Physicochemical Analyses of Fruits and Cassava. 1st ed. Cruz das Almas: Embrapa Cassava and Fruit Crops, 2010. p.

[009] As acerolas ripen, the vitamin C content decreases due to the oxidation of ascorbic acid caused by the exposure of the fruits to oxygen, light and heat (MACIEL, MIS, et al. Physicochemical characterization of fruits of acerola genotypes (Malpighia emarginata DC). Revista Ciência e Tecnologia de Alimentos, v. 30, n. 4, p. 865-869, 2010; PONTES, ATAC et al. Use of the acerola phenological cycle for standardization of the mechanized harvest point. In: XLIV Brazilian Congress of Agricultural Engineering. 2015, São Pedro / SP, 2015). This oxidation begins immediately after the fruit is harvested, through the action of enzymes such as ascorbinase or peroxidase, within the fruit itself (BRESOLIN, JD; HUBINGER, SZ. Methodology for the determination of ascorbic acid in citrus juices using high-performance liquid chromatography. In: III National Symposium on Agricultural Instrumentation. 2014. São Carlos / SP, 2014).

[010] It also contains carotenoids, such as beta-carotene, which corresponds to about 90% of the total carotenoids, in addition to beta-cryptoxanthin, phytofluene (PENHA, 2000), violaxanthin and lutein (ROSSO, VV. Composition of carotenoids and anthocyanins in acerola. Stability and antioxidant activity in model systems of anthocyanin extracts of acerola and açaí. 2006. 154 p. Thesis (Doctorate in Food Science) - Faculty Petition 870260037234, dated 04 / 22 / 2026, page 14 / 41 4 / 29 of Food Engineering, State University of Campinas, Campinas, 2006), which, together with ascorbic acid, give acerola antioxidant activity.

[011] Acerola is a climacteric fruit, with a high peak respiration rate, but a low peak ethylene production rate. It shows a marked increase in respiration rate during the ripening phase, accompanied by loss of firmness, color change, and development of flavor and aroma. (MACIEL, MIS, et al. Physicochemical characterization of fruits of acerola genotypes (Malpighia emarginata DC). Revista Ciência e Tecnologia de Alimentos, v. 30, n. 4, p. 865-869, 2010).

[012] Normally this respiratory peak occurs during the phase of change in peel pigmentation, from yellow to red. The climacteric pattern of respiration in acerola is accompanied by an increase in the synthesis and action of ethylene, accelerating its ripening and deterioration (RITZINGER, R.; RITZINGER, CHSP Acerola. Informe Agropecuário, v. 32, n. 264, p. 17-25, 2011). This rapid ripening and senescence of the fruits makes handling, storage and preservation difficult after harvest.

[013] Another factor that interferes with the shelf life of acerola fruits is the fact that they have a delicate skin, which is sensitive to light mechanical damage (with the rupture of the skin, the exposed pulp deteriorates rapidly). Post-harvest loss of national acerola production is estimated at around 40% (FREITAS, CAS et al. Acerola: production, composition, nutritional aspects and products. Revista Brasileira de Agrociências, v. 12, n. 4, p. 395-400, 2006). In addition to ripening, processing, mechanical damage, and temperature variations can degrade the vitamin C, carotenoid, and anthocyanin content of acerola cherries (PENHA, EM. Production of an acerola liqueur. 2000. 155 p. Thesis (Doctorate in Food Technology) Faculty of Food Engineering, State University of Campinas, Campinas, 2000; SILVA, WS. Quality and activity). Petition 870260037234, dated 04 / 22 / 2026, page 15 / 41 5 / 29 Antioxidant in fruits of acerola varieties. 2008. 137 p. Dissertation (Master's in Food Technology) - Faculty of Food Technology, Federal University of Ceará, Fortaleza, 2008).

[014] Because it is the most thermolabile vitamin, the vitamin C content may indicate that the other nutrients present in the food are possibly being preserved (ALVES, JA Kinetics of vitamin c degradation in minimally processed 'Palmer' mangoes stored at different temperatures. Ciência e Agrotecnologia, v. 34, n. 3, p. 714-721, 2010).

[015] Thus, the ascorbic acid content in fruits can serve as an indicator of ripeness, nutritional quality and preservation of the same (storage conditions, transport, etc.) (AZEREDO, HMC; BRITO, ES; GARUTTI. DS Chemical changes in food during storage. In: AZEREDO, HM Fundamentals of Food Stability. 2nd ed. Brasília: Embrapa, 2012. p. 41-75).

[016] Acerolas deteriorate rapidly when stored at room temperature, with a post-harvest shelf life of about 3 days when stored between 20 and 30°C (EMBRAPA. A Cultura da Acerola. Brasília: Embrapa, 2012. 150 p.).

[017] Thus, it is necessary to use preservation methods that increase their shelf life and maintain their organoleptic and nutritional characteristics for longer (MACIEL, MIS, et al. Physicochemical characterization of fruits of acerola genotypes (Malpighia emarginata DC). Revista Ciência e Tecnologia de Alimentos, v. 30, n. 4, p. 865-869, 2010). When stored at 8°C and packaged in flexible PVC film, acerolas can be preserved for up to 7 days, with up to 2% weight loss. At lower temperatures (between 5 and 8°C), the shelf life of the fruits increases to up to 10 days. In the case of fruits for export or for long-distance transport, storage at temperatures below -20°C (freezing) becomes the only viable form of fruit preservation (MACIEL, MIS et al. Petition 870260037234, dated 04 / 22 / 2026, p. 16 / 41 6 / 29 Post-harvest modifications in fruits of 16 acerola genotypes stored under refrigeration. Brazilian Journal of Agricultural Sciences, v. 3, n. 2, p. 157-163, 2008).

[018] However, the use of very low temperatures can cause damage to the fruit, such as discoloration of the peel, loss of texture (softening) and loss of shine (RITZINGER, R.; RITZINGER, CHSP Acerola. Informe Agropecuário, v. 32, n. 264, p. 17-25, 2011).

[019] Relative humidity also interferes with the characteristics of acerolas during storage. When kept at relative humidity below 85%, the fruits may exhibit wrinkling and loss of shine, rapid weight loss, and susceptibility to pathogens (MAZARO, SM et al. Postharvest quality of acerolas treated with salicylic acid. Revista Brasileira de Ciências Agrárias, v. 10, n. 4, p. 512-517, 2015.).

[020] In this way, alternatives that allow for the fresh consumption of perishable fruits (through increased shelf life) can provide greater use of their nutritional properties, favor direct trade by producers, given that a large part of the current production (about 60%) is destined for industries, and reduce the environmental impact generated by their processing. Biofilms for Use in Food

[021] Biofilms or edible films are flexible coatings with varied composition, formed from biological macromolecules, capable of producing continuous matrices with high cohesion, and used for coating foods, mainly those with high perishability, such as fruits and vegetables (OLIVEIRA, TA et al. Effect of tomato coating with biofilm on appearance and mass loss during storage. Revista Verde, v. 6, n. 1, p. 230-234, 2011).

[022] Biofilms made from hydrophobic macromolecules, such as lipids, have a good barrier to water vapor, but they have Petition 870260037234, dated 04 / 22 / 2026, page 17 / 41 7 / 29 undesirable mechanical properties (MULLER, PS Development of biodegradable active packaging from pine nut and cassava starch with natural antioxidants and antimicrobials for the preservation of organic butter. 2016. 178 p. Thesis (Doctorate in Food Engineering), Federal University of Paraná, Curitiba, 2016).

[023] Biofilms made from hydrophilic macromolecules, such as polysaccharides and proteins, exhibit reasonable barrier properties to water vapor, carbon dioxide, and oxygen under low relative humidity conditions, in addition to high mechanical strength and flexibility (FAKHOURI, FM et al. Edible films and coatings based on native starches and gelatin in the preservation and sensory acceptance of Crimson grapes. Revista Ciência e Tecnologia de Alimentos, v. 27, n. 2, p. 369-375, 2007; THOMAS, AB Physicochemical, microbiological quality and bioactive compounds of strawberries coated with cassava starch and propolis. 2016. 106 f. Dissertation (Master's in Food Science), Federal University of Lavras, Lavras, 2016).

[024] When applied to the surface of food, the biofilm forms a protective layer that reduces gas exchange, water loss and, consequently, the loss of nutritional value, thus increasing the product's shelf life. This oxygen barrier also reduces the volatilization of aromas and flavors in the food, preserving its organoleptic characteristics for longer, and reducing the oxidation of lipids, vitamins and pigments. (FAKHOURI, FM et al. Edible films and coatings based on native starches and gelatin in the preservation and sensory acceptance of Crimson grapes. Revista Ciência e Tecnologia de Alimentos, v. 27, n. 2, p. 369-375, 2007; THOMAS, AB Physicochemical, microbiological quality and bioactive compounds of strawberries coated with cassava starch and propolis. 2016. 106 f. Dissertation (Master's in Food Science), Federal University of Lavras, Lavras, 2016). An ideal coating should provide the food with... Petition 870260037234, dated 04 / 22 / 2026, p. 18 / 41 8 / 29 Coated coatings offer shine, an attractive appearance, and reduced weight loss by reducing normal respiration without inducing anaerobic conditions. Edible biofilms must be made with raw materials that are safe for human consumption, also known as GRAS (generally recognized as safe), according to FDA (Food and Drug Administration) regulations (AZEREDO, HM C; FARIA, JA; BRITO, ES Packaging and its interactions with food. 2012). Therefore, they do not pose a health risk when consumed. Another fundamental factor is their inertness in relation to the sensory characteristics of the foods to be coated, and that these foods have neutral color, odor, and taste, as well as adequate adhesion to the product surface, thus providing the desired protection (FAKHOURI et al. Edible films and coatings based on native starches and gelatin in the preservation and sensory acceptance of Crimson grapes. 2007).

[025] The use of biofilms to increase the shelf life of food dates back to the 12th century in China, when wax was applied to the surface of citrus fruits to preserve them during sea voyages (LUVIELMO, MM; LAMAS, SV Edible coatings on fruits. Technological Studies in Engineering, v. 8, n. 1, p. 8-15, 2013). This coating technique is still used today on fruits such as apples, for example.

[026] In the early 1930s, waxes of vegetable (carnauba), mineral (paraffin) and animal (beeswax) origin began to be used for fruit preservation (VILLADIEGO, AMD et al. Edible films and coatings in the preservation of food products. Revista Ceres, v. 52, n. 300, p. 221-224, 2005). From the 1960s onwards, the use of water-soluble polysaccharides became the most widely used and studied commercial option for increasing the shelf life of fresh foods.

[027] In recent years, several studies have been testing the use of biofilms and edible films in food, mainly aiming to increase shelf life and improve sensory characteristics (FERNÁNDEZ, NM et Petition 870260037234, dated 04 / 22 / 2026, page 19 / 41 9 / 29 al. Current state of the use of edible coatings on fruits and vegetables. Biotechnology in the Agricultural and Agroindustrial Sector Journal, v. 15, n. 2, p. 134-141, 2017). However, environmental and economic aspects have also been carefully considered, since in this case the use of packaging directly in contact with the food is dispensed with, thus reducing the generation of synthetic (non-biodegradable) packaging waste and, consequently, the cost associated with them (FAKHOURI, FM et al. Edible films and coatings based on native starches and gelatin in the preservation and sensory acceptance of Crimson grapes. Food Science and Technology Journal, v. 27, n. 2, p. 369-375.Development of biodegradable active packaging made from pine nut and cassava starch with natural antioxidants and antimicrobials for the preservation of organic butter. 2016. 178 p. Thesis (Doctorate in Food Engineering), Federal University of Paraná, Curitiba, 2016.

[028] One of the most widely used methods for obtaining biofilms is based on the preparation of a colloidal solution containing the macromolecule and other components (additives) in a solvent, which is usually water, a process called casting (BATISTA, JA Development, characterization and applications of biofilms based on pectin, gelatin and fatty acids in bananas and broccoli seeds. 2004. 140 p. Dissertation (Master's in Food and Nutrition) - Faculty of Food Engineering, State University of Campinas, Campinas, 2004). According to Rodrigues (2015), obtaining biofilms by casting begins with the formation of the gel through intra- and intermolecular cross-links between the polymer chains, which forms a semi-rigid three-dimensional matrix that surrounds and immobilizes the solvent used. Petition 870260037234, dated 04 / 22 / 2026, p. 20 / 41 10 / 29

[029] After solubilization, the solution is dried until the film is obtained, which will later be used in the coating (in this case called a film, an independent structure subsequently used as packaging); or the food is coated directly with the film-forming solution, and drying occurs on the surface of the product (in this case called a coating, covering or coating) (FALGUERA, V. et al. Edible films and coatings: Structures, active functions and trends in their use. Food Science & Technology, v. 22, p. 292-303, 2011).

[030] There are also simple precipitation processes (precipitation or phase change of the solution by evaporation of the solvent or by adding another solvent incompatible with the matrix); compound precipitation (mixture of two film-forming solutions containing matrices of opposite charges, generating precipitation of the polymer complex); and thermal gelation (sol-gel transition due to heating and consequent denaturation and precipitation of certain proteins) (BATISTA, JA Development, characterization and applications of biofilms based on pectin, gelatin and fatty acids in bananas and broccoli seeds. 2004. 140 p. Dissertation (Master's in Food and Nutrition) - Faculty of Food Engineering, State University of Campinas, Campinas, 2004).

[031] Edible biofilms can have various formulations, most of which are made up of biomolecules, such as proteins and polysaccharides. The basic constituents for preparing a film-forming solution are: a high molecular weight polymer (forming agent), such as proteins, polysaccharides or lipids; a solvent, which is usually water; and a plasticizing agent (BATISTA, JA Development, characterization and applications of biofilms based on pectin, gelatin and fatty acids in bananas and broccoli seeds. 2004. 140 p. Dissertation (Master's in Food and Nutrition) Faculty of Food Engineering, State University of Campinas, Campinas, 2004). The use of the various components in obtaining the Petition 870260037234, dated 04 / 22 / 2026, page 21 / 41 11 / 29 biofilm aims to improve its mechanical and physical properties. Various components (of natural or synthetic origin) can be added that interact with the food and increase its stability or functionality. Some studies have tested the addition of antimicrobial agents, antioxidants, and preservatives to biofilm formulations (ALVES et al. Quality of strawberries coated with antimicrobial edible coating based on different starch sources. 2011; COSTA. SS. Cassava starch and glycerol films, reinforced with nanocellulose and activated with red propolis. 2013; TORLAK, E.; SERT, D. Antibacterial effectiveness of chitosan-propolis coated polypropylene films against foodborne pathogens. 2013; BITTANTE, AMQB et al. Application of natural antimicrobial compounds in biofilms for food packaging. 2014; SILVEIRA, PTS et al. Post-harvest quality of gherkin (Cucumis anguria L.).) coated with corn starch with added propolis extract. 2015; SIRIPATRAWAN, U.; VITCHAYAKITTI, W. Improving functional properties of chitosan films as active food packaging by incorporating with propolis. 2016).

[032] One natural additive option for edible coatings is red propolis, discovered about 10 years ago and whose botanical origin is Dalbergia ecastophyllum, also known as rabo de bugio, a plant characteristic of the mangrove region of northeastern Brazil (SILVA, BB Characterization of red propolis: its botanical origin and the seasonal effect on its chemical composition and biological activity. 2008). Propolis

[033] Propolis is a complex mixture of balsamic, resinous, and gummy substances, with varying color and texture. These substances are collected by honeybees from various parts of plants, such as flowers, buds, and also resinous exudates (PINTO, LMA; PRADO, NRT; CARVALHO, LB Properties, uses and applications of propolis. Electronic Journal of Petition 870260037234, dated 04 / 22 / 2026, p. 22 / 41 12 / 29 Pharmacy, v. 8, n. 3, p. 76-100, 2011). This mixture of collected materials is altered by the bees, which add salivary secretions, pollen, and wax to it.

[034] In the beehive, propolis has several purposes, such as protecting the nest; sanitizing the combs and inner walls; covering dead animals that could not be removed from the colony, preventing their decomposition; and sealing openings (Embrapa. Propolis Production. Teresina: Embrapa, 2010. 02 p), protecting the animals from external temperature variations.

[035] The use of propolis as a folk remedy is quite ancient. It has diverse therapeutic properties, with its antibacterial and healing properties being the most attractive (PEREIRA, DS et al. History and main uses of bee propolis. Revista Agropecuária Científica no Semiárido, v. 11, n. 2, p. 01-21, 2015). It was widely used in South Africa during the war that occurred at the end of the 19th century, mainly due to its healing properties (PEREIRA, AS; SEIXAS, FRMS; NETO, FRA Propolis: 100 years of research and its future perspectives. Revista Química Nova, v. 25, n. 2, p. 321-326, 2002). In ancient Egypt, it was used to embalm corpses due to its antiputrefactive properties; in ancient Rome and Greece it was recognized for its medicinal properties (PEREIRA et al., 2015).

[036] In Brazil, propolis was and continues to be widely used in folk medicine. However, there was a growth in interest and use in the 1980s, with the publication of the work of Ernesto Ulrich Breyer, in which he described the therapeutic and antibiotic properties of this compound (SANTOS, JR Bioprospecting of geopropolis from Melipona fasciculata Smith. 2010. 82 p. Dissertation (Master's in Health Sciences), Federal University of Maranhão, São Luís, 2010). Several studies have demonstrated the various properties of propolis, such as antioxidant (ALVES, E.; KUBOTA, EH Content of phenolics, total flavonoids and antioxidant activity of Petition 870260037234, dated 04 / 22 / 2026, page 23 / 41 13 / 29 samples of commercial propolis. Journal of Basic and Applied Pharmaceutical Sciences, v. 34, n. 1, p. 37-41, 2013; DE-MELO, AAM et al. Antioxidant capacity of propolis. Journal of Tropical Agricultural Research, v. 44, n. 3, p. 341-348, 2014; KUNRATH, CA Application and evaluation of propolis, the natural antioxidant in Italian-type salami. Brazilian Journal of Food Technology, v. 20, p. 1-10, 2017.), antibacterial (MARCUCCI, MC; GUTIERREZ-GONÇALVES, MEJ. Antimicrobial and antioxidant activities of propolis from the state of Ceará. Revista Fitos, v. 4, n. 1, p. 81-86, 2009; CAMPOS, VAC et al. Antibacterial activity of propolis produced by Frieseomelitta varia. Ciência e Agrotecnologia, v. 35, n. 6, p. 1043-1049, 2011; ANDRADE, NPC et al. In vitro antimicrobial activity of ethanolic extracts of propolis from three Brazilian states on Aeromonas hydrophila isolated from fish. Arquivos do Instituto Biológico, v. 79, n. 1, p. 9-15, 2012; SIQUEIRA, ALet al. Study of antibacterial action of hydroalcoholic extract of propolis red on Enterococcus faecalis. UNESP Dentistry Magazine, v. 43, no. 6, p. 359366, 2014), anti-inflammatory (REIS, CMF et al. Anti-inflammatory, anti-gastric ulcer activity and subchronic toxicity of ethanolic propolis extract. Brazilian Journal of Pharmacognosy, v. 9, n. 10, p. 43-52, 2000; ALBUQUERQUE-JUNIOR, RLC et al. Effect of bovine type-I collagen-based films containing red propolis on dermal wound healing in rodent model. International Journal of Morphology, v. 27, n. 4, p. 1105-1110, 2009; VEGA, DF et al. Effect of treatment with oral red propolis extract in non-alcoholic steatohepatitis. Cuban Journal of Medicine, v. 53, n. 3, p. 282-290, 2014), among others.

[037] The chemical composition of propolis consists of approximately 50% plant resin, 30% wax, 10% essential oils, 5% pollen, and 5% various substances (BOGDANOV, S. Propolis: Composition, Health, Medicine: A Review. Bee Product Science, 2017). It may also contain, Petition 870260037234, dated 04 / 22 / 2026, page 24 / 41 14 / 29 minerals, vitamins (B1, B2, B3 and B6), lactones, quinones, steroids, sugars and natural pigments, such as carotenoids and chlorophyll (THOMAS, AB Physicochemical, microbiological quality and bioactive compounds of strawberries coated with cassava starch and propolis. 2016. 106 p. Dissertation (Master's in Food Science), Federal University of Lavras, Lavras, 2016).

[038] Brazil, due to the enormous variability of its flora, presents several types of propolis, considering that its composition varies according to the botanical source from which the bees extracted the resin and also with some factors, such as the collection period, the region's climate, among others. The biological potential of each type of propolis is due to the synergism between its many constituents (LUSTOSA, SR et al. Propolis: updates on chemistry and pharmacology. Brazilian Journal of Pharmacognosy, v. 18, n. 3, p. 447-454, 2008).

[039] The variety of Brazilian propolis makes them products with enormous technological potential, mainly due to their very specific geographical and botanical origin from certain regions of the country. Red Propolis

[040] Red propolis is the 13th type of propolis discovered in Brazil. Its botanical origin is Dalbergia ecastophyllum, also known as “rabo de bugio”, a plant characteristic of the mangrove regions of northeastern Brazil, mainly in the states of Alagoas, Bahia and Paraíba. Bees collect the exudate from the plant's stems and, from this, produce red propolis.

[041] In 2012, Alagoas red propolis received Geographical Indication certification in the Designation of Origin modality, provided by the National Institute of Industrial Property.

[042] This type of propolis has a phytochemical composition similar to a type of red propolis produced in Cuba. However, Cuban propolis does not contain benzophenones (PICCINELLI, AL et al. Isoflavonoids Isolated from Petition 870260037234, dated 04 / 22 / 2026, page 25 / 41 15 / 29 Cuban Propolis. Journal of Agricultural and Food Chemistry, v. 53, p. 9010-9016, 2005), which are found in Alagoas red propolis. (TRUSHEVA, B. et al. Bioactive constituents of Brazilian red propolis. Evidence-Based Complementary and Alternative Medicine, v. 3, n. 2, p. 249-254, 2006) identified compounds with antibiotic, antifungal, antioxidant, and cytotoxic activity in samples of red propolis.

[043] In a study conducted by (ALENCAR, SM et al. Chemical composition and biological activity of a new type of Brazilian propolis: Red propolis. Journal of Ethnopharmacology. v. 113, n. 2, p. 278-283, 2007), bioactive compounds (isoflavonoids, chalcones and benzophenones) were identified in the red propolis from Alagoas, compounds that had not been reported in any of the other 12 types of Brazilian propolis nor in the red propolis from Cuba, in addition to also observing antimicrobial activity.

[044] The presence of various flavonoids and phenolic acids in propolis extracts and tinctures shows that these substances act synergistically in cytostatic / cytotoxic, anti-inflammatory, healing, antimicrobial, and antioxidant actions (Freires, IA; de Alencar, SM; Rosalen, PL. A pharmacological perspective on the use of Brazilian red propolis and its isolated compounds against human diseases. European Journal of Medicinal Chemistry v. 110, p. 267-279, 2016). Thus, the cocktail of combined substances, even at low concentrations, will promote a potent action against pathogenic agents, especially in the wound healing process. (SILVA, BB. Characterization of red propolis: its botanical origin and the seasonal effect on its chemical composition and biological activity. 2008. 51 p. Dissertation (Master's in Dentistry) - Faculty of Dentistry, State University of Campinas, Piracicaba, 2008.) observed high antimicrobial activity of red propolis extract against Streptococcus mutans, Streptococcus sobrinus, Staphylococcus aureus and Actinomyces naeslundii. Petition 870260037234, dated 04 / 22 / 2026, page 26 / 41 16 / 29

[045] Applications for edible coatings with diverse formulations and purposes were verified: increasing the functional properties of fruits and vegetables (PI1102766-5A2); preserving frozen bakery products (BR1120150174647A2 and BR1120160169646A2); increasing the shelf life of food in general (PI1104166-8A2, BR1120180107670A2, PI88007618A2, BR1020170073653A2).

[046] However, none of the patent documents described uses red propolis as an additive in edible coatings and biofilms.

[047] Patent application BR1020180091980 deals with a membrane (dressing, drying the membrane before use) containing red propolis for application in the treatment of wounds. The present invention uses the formulation of the cited application. However, the formulation has been diluted and used in the form of a film-forming solution (drying after application) for coating fruits and vegetables. Thus, the application and use are different from the cited patent.

[048] The developed biofilm uses pectin and carboxymethylcellulose as a base, components of plant origin that are widely used in the food industry as thickeners and stabilizers (CALEGUER, VF; BENASSI, MT Effect of the addition of pulp, carboxymethylcellulose and gum arabic on the sensory characteristics and acceptance of powdered preparations for orange-flavored soft drinks. 2007; CANTERI, MHG et al. Pectin: From Raw Material to Final Product. 2012). Some positive characteristics of using pectin and carboxymethylcellulose in edible biofilms are their high water retention capacity, low cost, inertness in relation to the applied food, in addition to being renewable and biodegradable. SUMMARY OF THE INVENTION

[049] The present invention discloses red propolis biofilms, composed of crude hydroalcoholic extract of red propolis, thickening agent, humectant, preservative, plasticizer, stabilizing agent and dispersant or emulsifier. The invention also deals with the process of obtaining the biofilms, Petition 870260037234, dated 04 / 22 / 2026, page 27 / 41 17 / 29 by means of the casting technique. The disclosed composition comprises a polymeric matrix containing active substance(s) of red propolis combined with pharmaceutical excipients. Furthermore, the invention relates to the process of preparing the composition by performing physical mixtures with red propolis to ensure content and uniformity in the mixture. Finally, uses for biofilms and for the composition in question are presented. BRIEF DESCRIPTION OF THE FIGURES / DRAWINGS

[050] The attached Figure 1 presents a flowchart of the process for obtaining film-forming solutions and coatings for fruits and vegetables.

[051] The attached Figure 2 shows the evaluation of the pH increase of coated and uncoated fruits over 15 days of storage.

[052] The attached Figure 3 shows the determination of vitamin C loss from coated and uncoated fruits over 15 days of storage.

[053] The attached Figure 4 presents the evaluation of mass loss of coated and uncoated fruits over 15 days of storage.

[054] The attached Figure 5 shows the increase in soluble solids content of coated and uncoated fruits over 15 days of storage.

[055] The attached Figure 6 shows the decrease in total titratable acidity of coated and uncoated fruits over 15 days of storage.

[056] The attached Figure 7 shows the increase in the degree of ripeness of coated and uncoated fruits over 15 days of storage. DETAILED DESCRIPTION OF THE INVENTION

[057] The products, processes and composition described in the present invention can be better detailed and understood through the figures in this document and the following description: Preparation of Hydroalcoholic Extract of Red Propolis (EHPV)

[058] A 300 g sample of propolis was subjected to an ultrasonic extraction process with a power of 100 W for 30 minutes, in the range of 500 to 1000 mL of 50 to 99 °GL alcohol, preferably between 70 and 80 °GL, with three repetitions. Petition 870260037234, dated 04 / 22 / 2026, page 28 / 41 18 / 29 of this same process. The entire extraction always took place in a dark environment, due to the photosensitivity of the sample. The extract was then filtered and stored in an amber glass bottle for better preservation. After complete extraction of the active substances, the material is concentrated in a rotary evaporator, coupled to a vacuum pump, using a rotation speed of 20 to 120 rpm, in a water bath at a temperature range of 35 to 55°C and a pressure between 400 and 700 mm Hg. Alternatively, it can be concentrated using a water bath at 37°C to 42°C.

[059] A dark solid mass is obtained with a solvent percentage between 5 and 35%, and is called crude red propolis extract. The crude extract should preferably have a solvent percentage between 0.5 and 12%. Preparation of biofilms loaded with red propolis (BCPV)

[060] The biofilms presented in this document exhibit concentrations of phenolic compounds such as flavonoids, isoflavonoids, phenolic acids, as well as chalcones due to the specific characteristics of red propolis. In addition, the biofilms in question exhibit relevant characteristics of protecting fruits and vegetables from pro-oxidant action through the phenolic compounds present in the red propolis extract. Furthermore, the biofilm promotes a reduction in gas exchange and helps reduce enzymatic browning rates, given that oxygen, in contact with the polyphenol oxidase enzymes present in the food, reacts with them, forming melanins (dark pigments), which leads to rejection by consumers.

[061] In one embodiment of the invention, the biofilm loaded with red propolis extract presented in this invention can be applied to fruits and vegetables of different sizes and plant anatomical shapes. The biofilm, in addition to the extract with the active principles of interest, obtained from red propolis, has in its composition pharmaceutical excipients with different functionalities, namely: active compound (red propolis extract), Petition 870260037234, dated 04 / 22 / 2026, pp. 29 / 41 19 / 29 film-forming polymeric matrix with coating, emulsifying, preservative, stabilizing and antioxidant functions.

[062] In one embodiment of the present invention, BCPV is presented as a composition of: standardized hydroalcoholic extract of red propolis derived from crude red propolis in a proportion between 0.1 and 6%, preferably between 1 and 5%; thickening agent in a proportion between 0.1 and 5%, preferably between 1 and 3%; dispersant in a proportion between 5 and 95%; humectant in a proportion between 0.1 and 5%, preferably between 0.5 and 2%; stabilizing agent in a proportion between 1 and 5%; and preservative in a proportion between 0.01 and 0.05%.

[063] Red propolis biofilms (RPBs), due to their composition containing carboxymethylcellulose excipients, exhibit thickening properties, producing viscosity and stabilizing / binding the components of the formula, consequently avoiding variations in the uniformity of the RPB content.

[064] It is important to highlight that the biofilms proposed in this document are innovative, as they contain red propolis extract in their composition (0.1 to 6%), which is not found in any of the patents available in patent search databases for biofilms for coating foods such as fruits and vegetables.

[065] The steps for obtaining BCPV include preparing the crude extract from red propolis, preparing the physical mixture of the formula components with EEPV, coating the fruit or vegetable and drying at room temperature (Figure 1). Preparation of the physical mixture containing red propolis.

[066] The formulations were made using 1 to 3% sodium carboxymethylcellulose (NaCMC) hydrogel, which was prepared beforehand. To prepare this gel, the NaCMC powder was weighed on an analytical balance and then transferred to a beaker, along with a given volume of distilled water previously measured in a graduated cylinder. With the aid of a glass rod, the Petition 870260037234, dated 04 / 22 / 2026, pp. 30 / 41 20 / 29 of the contents were mixed and then subjected to an ultrasound device for a period ranging from 10 to 30 minutes for better homogenization. Then, the remaining components of the formula were added, such as citrus pectin, propylene glycol, and 70% sorbitol, always with agitation in a mechanical mixer with a rotation speed ranging from 200 to 2000 rpm. During the homogenization process, the ethanolic extract of red propolis (EEPV) was incorporated into the formulation in proportions ranging from 0.5 to 6%. Fruit coating process

[067] The acerolas were harvested in the morning, packed in thermal boxes containing ice and transported immediately for coating. The fruits were selected according to the degree of ripeness, based on the color of the peel (reddish-orange), the ripening point recommended by EMBRAPA (2016) for harvesting fruits intended for fresh consumption (21 to 23 days after anthesis).

[068] After selection, the fruits were sanitized by immersion in a 100 ppm sodium hypochlorite solution for 15 minutes. They were then washed with running water, drained, and dried at room temperature. The fruits were coated by immersing them in a biofilm solution loaded with red propolis. The immersion time ranged from 30 seconds to four minutes, preferably for one minute, followed by drying. In this biofilm method, drying occurred in different ways: one at room temperature (25°C and 60% humidity) and the other under refrigeration (5°C and 60% humidity) for up to 24 hours.

[069] The fruits were then drained and dried naturally on stainless steel racks, at room temperature (25°C and 60% air humidity) and under refrigeration (5°C and 60% air humidity). - Obtaining carnauba wax and coating the fruits

[070] Carnauba wax was used for comparison between the developed biofilm and the coating normally used by the fruit industry. A Petition 870260037234, dated 04 / 22 / 2026, pages 31 / 41 21 / 29 wax was purchased from supplier GM - Comércio de Ceras e Derivados LTDA. Type 3 wax, indicated for coating fruits, was used. A 1.5% wax-in-water emulsion was prepared for subsequent coating of the fruits by immersion and drying at room temperature and under refrigeration. - Characterization of Red Propolis Extract

[071] The red propolis extract, after the extraction process, was characterized using different analytical techniques to verify the presence of phenolic compounds, as well as to demonstrate in vitro antioxidant activity. Among the analytical techniques, we mention: A) Antioxidant activity using DPPH (2,2-diphenyl-1-picrylhydrazyl). A stock solution of 1 mg / mL of the red propolis extract was prepared in absolute ethanol. A 1 mM solution of the DPPH· radical in absolute ethanol was also prepared and stored in an amber glass container. Then, 2 mL of the DPPH· solution were added to 5 mL volumetric flasks and, at 1-minute intervals for each flask, a 125 µL aliquot of the EHPV sample was added and the volume was completed with absolute ethanol, resulting in a concentration of 25 μg / mL. The reaction was allowed to occur in a dark environment for 30 minutes.After this time, the samples were subjected to reading in a Shimadzu model 1240 UV-Vis spectrophotometer in photometric mode, adjusted to a wavelength of 517 nm. The blank test of the samples was obtained with an aliquot of the DPPH· solution and another with absolute ethanol. The percentage of remaining DPPH· radical, after 30 minutes, was calculated according to the following formula: (%) of Remaining DPPH· = [(Sample - Blank) / (Control - Blank)] x 100. Where: Sample: absorbance of the reaction between the DPPH· radical solution and the antioxidant sample; Blank: absorbance of the solvent solution used to prepare the antioxidant sample; Control: absorbance of the DPPH· radical with a small aliquot of the solvent used to prepare the sample, replacing the solution of the sample under study itself. Followed by the calculation of... Petition 870260037234, dated 04 / 22 / 2026, pages 32 / 41 22 / 29 Antioxidant activity: (%) inhibition of the DPPH radical = 100 - (%) remaining DPPH. B) Antioxidant power by the ferric reducing antioxidant power (FRAP) method. To 5 mL volumetric flasks, 90 pL aliquots containing 25 pg / mL of the EHPV sample, 270 pL of distilled water, and 2.7 mL of FRAP reagent (prepared from 25 mL of 0.3 M acetate buffer solution, 2.5 mL of 10 mM TPTZ (2,4,6-Tris(2-pyridyl)-striazine) solution, and 2.5 mL of 20 mM aqueous ferric chloride solution) were added. The solutions were homogenized in a tube shaker and kept in a water bath at 37 °C for 30 minutes. After the reaction time had elapsed, the sample was subjected to reading in a Shimadzu model 1240 UV-Vis spectrophotometer, adjusted to a wavelength of 595 nm. The FRAP reagent was used as a blank to calibrate the spectrophotometer. The ferrous sulfate calibration curve was used to determine the antioxidant power by iron reduction.An analytical calibration curve was constructed from an aqueous solution of Ferrous Sulfate Heptahydrate (FeSO4.7H2O) as a standard at increasing concentrations of 500, 750, 1000, 1250, 1500, and 2000 μM / mL. C) Determination of total phenols. The total phenol content was determined by UV-Vis spectrophotometry, using gallic acid as a standard, according to the Folin-Ciocaulteau method described by WOISKY (1996). To 5 mL volumetric flasks, 3.5 mL of distilled water, 400 μL of Folin-Ciocaulteau reagent, and the aliquot of gallic acid stock solution corresponding to each desired concentration (20, 30, 40, 50, 60, 70, 80, and 90 μg / mL) were added, and the mixture was shaken for a few seconds. Then, at intervals of 1 to 8 minutes, 600 μL of 20% sodium carbonate solution were added, and the volume was completed with distilled water. After 2 hours, triplicate readings were performed using a Shimadzu 1240 UV-Vis spectrophotometer, adjusting the wavelength to 760 nm.Finally, the equation of the line was calculated using the least squares method. For the determination of total phenols in EHPV, an aliquot with a concentration of 20 μg / mL was used, following the methodology already described. The linear coefficient of the curve... Petition 870260037234, dated 04 / 22 / 2026, pp. 33 / 41 The value obtained for 23 / 29 was 0.9971. D) Determination of total flavonoids. The total flavonoid content was determined by the aluminum chloride method, using quercetin as a standard. To 5 mL volumetric flasks, a 4 mL aliquot of methanol, 0.1 mL of 5% aluminum chloride solution, and the corresponding aliquot of quercetin stock solution were added for each of the desired concentrations (4, 6, 8, 10, 12, 14, and 16 μg / mL). The final volume of the flask was completed with methanol, shaking for a few seconds. The samples were kept in a dark environment for 30 minutes, after which the reading was taken on a Shimadzu model 1240 UV-Vis spectrophotometer, adjusting the wavelength to 425 nm. The equation of the line was then calculated using the least squares method. The resulting linear coefficient was 0.9932.To determine the flavonoid content in EHPV, an aliquot with a concentration of 200 μg / mL was used, employing the methodology described previously. - Characterization of Red Propolis Loaded Biofilms (BCPV)

[072] Red propolis loaded biofilms and carnauba wax biofilms were dried at 25°C on glass plates for 12 hours and then characterized using techniques such as: A) Moisture determination. The moisture content of the dried biofilms was determined by an infrared drying and moisture removal method. Samples of the dried biofilms were cut into 4x4 diameter pieces, weighing 0.5g. They were then analyzed in a Shimadzu MOC63u infrared moisture analyzer at 105°C for 10 minutes. The samples were analyzed in triplicate. B) Water Solubility. The water solubility of the dried biofilms was determined according to the following methodology. Samples of dried biofilms were cut into 1cm discs, weighed, immersed in distilled water, and kept under mechanical agitation for 24 hours at 25°C.After this period, the samples were dried in an oven at 105°C for 24 hours and weighed again when they were ready. Petition 870260037234, dated 04 / 22 / 2026, pp. 34 / 41 24 / 29 completely cold, thus determining the final dry mass of the samples. The analysis was performed in triplicate. Solubility was expressed in terms of dissolved dry mass, calculated according to the equation: Sol = (mi mf) / mi X 100. Where, Sol: solubility in water (g / 100g of film); mi: initial mass of the sample (g); mf: final dry mass of the sample (g) after solubilization. C) Mass loss. The mass loss of the fruits was determined by weighing on a Shimadzu analytical balance, model AUY 220. Then, the percentage of mass loss of the fruits was calculated. The analysis was performed in triplicate. D) Total Titratable Acidity (TTA). TTA was determined using the volumetric methodology with phenolphthalein indicator and titration with sodium hydroxide, according to the standards of the Adolfo Lutz Institute (IAL, 2008). The analysis was performed in triplicate. The percentage of ATT was calculated using the following formula: ATT = ((V x N xfx F x 100) / (P)).Where ATT: total titratable acidity in % of citric acid per 100g of pulp; V: volume of NaOH used in the titration in ml; N: normality of the NaOH solution; f: correction factor of the NaOH solution used; F: factor of the predominant acid in the fruit (citric acid); P: weight or volume of the sample used. E) Vitamin C. Vitamin C determination was performed by spectrophotometry, according to the methodology described by Oliveira (2010). A Shimadzu model 1240 UV-Vis spectrophotometer was used. F) pH. The pH of the fruits was determined by direct reading using a Quimis model Q400AS benchtop potentiometer. The samples were analyzed in triplicate. Uses of Biofilms Loaded with Red Propolis

[073] The biological activities of red propolis are already well described in the literature and biofilms loaded with red propolis can be used in a wide range of industrial sectors, cosmetics, pharmaceuticals and especially food.

[074] The use of propolis as an active antioxidant, anti-inflammatory, healing, preservative and other active ingredient is already well documented in Petition 870260037234, dated 04 / 22 / 2026, pages 35 / 41 25 / 29 literature. However, red propolis has a different chemical composition from other types of propolis already described in the state of the art. Thus, it is a strong indication of varied biological activities.

[075] The biofilm compositions presented here demonstrate antioxidant and preservative activities performed by the in vitro DPPH and FRAP antioxidant assays using the hydroalcoholic extract of red propolis. The biological activities, such as the antioxidant activity of the extract present in the biofilm compositions, duly proven by the proposed processes, can be used for: edible coating of perishable climacteric fruits and coating of vegetables of various sizes, shapes, and ripening times. The coating of fruits has the capacity to increase the shelf life of fruits and vegetables at room temperature and refrigerated temperatures ranging from 4°C to 25°C. Furthermore, it allows for greater stability, reducing impacts regarding mass loss, pH, acidity, moisture, and especially greater macroscopic and morphological control of fruits and vegetables.

[076] The film-forming compositions of biofilms loaded with red propolis extract are edible and provide phenolic compounds, classified as phenols and flavonoids, which are directly linked to the biological activities of propolis acting against free radicals. In addition, it protects fruits and vegetables against oxidative processes.

[077] Thus, the embodiments of the invention described in this document represent an advance in the state of the art, since they allow the production of edible biofilms loaded with red propolis extract, obtained through standardized processes that characterize safe and economically viable production because they are composed of low-cost natural polymers. In this way, the BCPV obtained through the proposed process can be used for their biological activities and the action of red propolis. Petition 870260037234, dated 04 / 22 / 2026, pp. 36 / 41 26 / 29 as a product in various industrial sectors in the cosmetic and pharmaceutical areas, and especially in the food industry. EXAMPLES

[078] The BCPV evaluations obtained using the proposed process demonstrated the production of a suitable, pure product with preserved activity. The results obtained are shown in the tables and figures indicated.

[079] Table 1 presents the evaluation of the antioxidant activity of EHPV using the DPPH and FRAP methods. Table 1 Methodology Sample Concentration Antioxidant Activity DPPH· FRAP 25 pg / mL 25 pg / mL 92.89% < 0.00 57.4 mMol of FeSO4 / g propolis ± 2.69

[080] EHPV showed a 92.89% antiradical effect, according to the DPPH· methodology. For the FRAP methodology, the result was 57.4 mMol of FeSO4 / g propolis. It is interesting to use more than one methodology to determine the antioxidant activity of propolis, considering that divergent values ​​are found in the literature, a fact that may be associated with differences in methodologies, as well as seasonality and different geographical and botanical origins. Thus, the results obtained in the present study show a high antioxidant capacity of EHPV.

[081] Table 2 presents the determination of total phenols and flavonoids of EHPV. Table 2 Petition 870260037234, dated 04 / 22 / 2026, pp. 37 / 41 27 / 29 Analysis of Sample Concentrations: Results (mg / g) Results (%) Total Phenols 20 pg / mL 243.20 mg ± 3.481 24.32% Total Flavonoids 200 pg / mL 37.80 mg ± 0.962 3.78% 1: mg EAG / g (expressed as gallic acid equivalent per g of red propolis); 2: mg EQ / g (expressed as quercetin equivalent per g of red propolis).

[082] The total phenol and flavonoid contents were 243.20 mg GAE / g and 37.80 mg QE / g, respectively. The literature describes the phenolic compound content in red propolis as quite variable, usually being greater than 90 mg / g. The total phenol content found in the present study is consistent with and corroborates existing studies. On the other hand, the flavonoid content of red propolis tends to be low compared to the total phenolic content. These results highlight the variability in flavonoid content found in the literature. The values ​​found by this study are similar to existing studies.

[083] EXAMPLE 1 - Evaluation of the pH of fruits coated with a biofilm loaded with red propolis.

[084] The use of biofilm maintained, throughout storage, a lower pH in coated fruits when compared to uncoated fruits (Figure 2). pH tends to increase during storage due to the degradation of organic acids and the action of enzymes. Furthermore, it is a useful parameter for determining the degree of ripeness and deterioration of fruits. Thus, a smaller increase in pH indicates that the coating may Petition 870260037234, dated 04 / 22 / 2026, pp. 38 / 41 28 / 29 preserve the nutritional characteristics of the fruits for longer, in addition to extending the post-harvest shelf life.

[085] EXAMPLE 2 - Determination of vitamin C loss in coated and uncoated fruits over 15 days of storage.

[086] Coating the fruits with biofilm also resulted in less vitamin C loss during storage (figure 3). Because it is the most heat-labile vitamin, the vitamin C content may indicate that other nutrients present in the food are possibly being preserved. Thus, the ascorbic acid content in fruits can serve as an indicator of ripeness, nutritional quality and preservation.

[087] EXAMPLE 3 - Evaluation of mass loss of coated and uncoated fruits over 15 days of storage.

[088] The use of biofilm also provided a reduction in fruit mass loss when compared to the control (uncoated fruit) (Figure 4). Mass loss in fruits is mainly caused by water loss to the environment in the form of vapor, which also leads to surface dehydration (wrinkling and wilting). Thus, less mass loss indicates a reduction in water loss during storage.

[089] EXAMPLE 4 - Increase in soluble solids content of coated and uncoated fruits over 15 days of storage.

[090] The coating provided a lower soluble solids content throughout storage (Figure 5). In fruits, soluble solids are mostly composed of sugars, and tend to increase with ripening. Thus, a lower soluble solids content indicates a lower degree of ripeness and the possibility of a longer post-harvest shelf life.

[091] EXAMPLE 5 - Decrease in total titratable acidity of coated and uncoated fruits over 15 days of storage.

[092] Total titratable acidity was also influenced by the coating (Figure 6). Fruits coated with the biofilm showed higher total acidity throughout Petition 870260037234, dated 04 / 22 / 2026, pp. 39 / 41 29 / 29 of storage compared to uncoated fruits. In fruits, acidity is mainly related to organic acids dissolved in the vacuoles of the cells. In addition to acidity, these acids contribute to the characteristic aroma of the food, and their content tends to decrease with ripening, as they are used as a substrate in the respiratory process or are converted into sugars. Thus, higher acidity indicates that it was possible to preserve the organic acids in the food, which implies a lower degree of ripeness and a longer shelf life.

[093] EXAMPLE 6 - Increase in the degree of ripeness of coated and uncoated fruits over 15 days of storage.

[094] As a consequence of the reduction in the formation of soluble solids and preservation of total acidity observed, a reduction in the degree of ripeness was verified in relation to the uncoated fruit (Figure 7), which indicates that the shelf life of the fruits can be extended. Petition 870260037234, dated 04 / 22 / 2026, pp. 40 / 41

Claims

Claims 1. Edible biofilm characterized by comprising: a) standardized hydroalcoholic extract of red propolis in a proportion of 0.1% to 6% by mass; b) thickening agent, comprising carboxymethylcellulose and pectin, in a total proportion of 0.1% to 5% by mass; c) humectant, comprising propylene glycol and sorbitol, in a total proportion of 0.1% to 5% by mass; d) preservative in a proportion of 0.01% to 0.05% by mass; e) stabilizing agent, comprising carboxymethylcellulose, in a total proportion of 1% to 5% by mass; f) dispersant, emulsifier, in a proportion of 5% to 95% by mass.

2. Edible biofilm, according to claim 1, characterized by the presence of a hydroalcoholic extract of red propolis in a proportion of 1% to 5% by mass.

3. Use of an edible biofilm based on red propolis, according to claim 1, characterized by being an edible coating for the preservation of fruits and vegetables.

4. Use of an edible biofilm based on red propolis, according to claim 1, characterized by being for topical use for cosmetic purposes. Petition 870260037234, dated 04 / 22 / 2026, pp. 41 / 41

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