Process for preparing a vegetable-based dye, composition comprising hydroalcoholic extracts and its use

Description

1 / 39 “PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN, COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS AND ITS USE FIELD OF THE INVENTION:

[001] The present invention discloses processes for preparing a dye of vegetable origin comprising hydroalcoholic extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris; and dye compositions comprising said hydroalcoholic extracts.

[002] The present invention falls within the field of pigments of natural origin, more specifically, in the area of ​​natural dyes for application in food, cosmetics, personal hygiene products and textiles. BACKGROUND OF THE INVENTION:

[003] Currently, there is a perceived revaluation of the use of natural inputs in various products for human use, seeking a more ecological, healthy approach that is less harmful to animals and humans. Along these lines, there has been a growing interest in natural products, essential oils, plant-based products, "cruelty-free" items, or those free of entirely synthetic compounds, so that these products have a low environmental impact and are more socially responsible.

[004] Within this context, dyes are also present, a class of products indispensable to various industries, such as cosmetics, textiles, and food. Dyes are substances that impart color to materials, being widely used in various industries, and the availability of dyes of natural origin enables the development of products that meet these consumer demands.

[005] Thus, the search for natural dyes has increased worldwide, especially in the food industry. In the cosmetics industry, for example, it is observed Petition 870250012114, dated 02 / 14 / 2025, page 9 / 60 2 / 39 It is known that, in the list of 153 colorants permitted for use by the European Union, approximately 8% are of vegetable origin, 20% are inorganic, and 72% are synthetic.

[006] Among the advantages of using dyes of natural origin, their biodegradable characteristics stand out, resulting in a lower environmental impact, as well as their reduced allergenicity and toxicity when compared to synthetic dyes. In addition to seeking dyes of natural origin, it is also important that they are obtained from plant sources, aiming to obtain a "cruelty-free" dye. For example, regarding the red dye of natural origin, the commonly used source is carmine dye, which is obtained by crushing the cochineal insect. This process, besides using harmful solvents such as chloroform, also has a controversial origin from environmental and social perspectives.

[007] Thus, natural dyes obtained from plants prove to be advantageous and may also present additional properties, such as antioxidant, antimicrobial, healing, pH regulating characteristics, among others, which provide health benefits.

[008] Plant-derived dyes commonly exhibit different chemical structures from synthetic compounds, which natural dyes are composed of carotenoids, polyphenols, quinones, or alkaloids. These structures give these dyes a lower degree of stability when exposed to differences in pH, temperature, or light, compared to dyes of synthetic origin. Because of this, research in the area of ​​natural dyes has focused on methodologies that seek to improve their stability, both in the free dye and in its formulation.

[009] In addition, the color of a product can also be influenced by its overall composition, through Petition 870250012114, dated 02 / 14 / 2025, page 10 / 60 3 / 39 incompatibility between components, which in the case of the cosmetic and food industries, should not suffer drastic changes in their properties when stored.

[0010] Several plants can be used as a source of natural pigments. Fridericia chica (formerly called Arrabiaea chica), also known as crajiru, pariri, cipó-cruz or crejer, is a shrubby, climbing or arboreal plant from the Bignoniaceae family that inhabits areas from Central America to southern Brazil, but can also be found in Africa. Among its interesting characteristics are its wide medicinal use against intestinal colic, anemia, its healing action, and its use against ulcers and other ailments. It can also be used as a body dye and tattoo ink of a reddish-brown color, mainly by indigenous peoples. Its pigment is called carajura or crajura.

[0011] Its therapeutic use, as a wound healer, antioxidant, anti-ulcerative, and anti-colic agent, is widely explored due to the low potential for adverse effects, both for topical and internal use. For example, patent PI 1105312-7 B1 addresses pharmaceutical compositions comprising Arrabidaea chica extract in micro- and nanoparticulate delivery systems and liposomes, as well as processes for obtaining and using them for the healing of skin lesions. Patent BRPI0600943B1 addresses the use of Arrabidaea chica extract as an antifungal and antibacterial agent in pharmaceuticals or additives in cosmetics, and patent BR102016028506B1 describes the preparation of polymeric membranes by electrospinning through the association of extracts of Pterodon pubescens Benth and Arrabidaea chica Verlot for application in guided bone and tissue regeneration, and also reveals the pharmacological activities of action Petition 870250012114, dated 02 / 14 / 2025, page 11 / 60 4 / 39 healing and antimicrobial with anti-inflammatory and analgesic action of the extracts.

[0012] However, its use as a possible raw material for natural dyes is not widely explored. Documents JP5186089B2, JP2008069124A, US8632612B2 and CN101332163B suggest crajiru as a source of natural pigments, but they do not specifically explore crajiru as a raw material for dye production, only mentioning it as a source of natural pigments.

[0013] An important factor when considering the use of natural dyes is their stability, especially when dealing with anthocyanins, which are highly susceptible to variations in pH, temperature and light – this being the most limiting factor for their industrial use.

[0014] Thus, one way to contain its degradation is through the use of co-pigments in its mixture, from the combination of different dyes. Such compounds can be flavonoids, polyphenols, alkaloids, organic acids, amino acids and even other anthocyanins, and give the pigments bathochromic (change of maximum λ) or hyperchromic (increases in absorbance of maximum λ) effects, also promoting an increase in their stability.

[0015] Thus, another interesting source of naturally occurring pigments is açaí (Euterpe oleracea Mart.). This fruit is widely used in food, especially in northeastern Brazil, and has proven to be very promising economically since investments in its production began in the 1990s. Currently, it is sought after due to its high energy and nutritional value, as it is a food with high levels of protein, fiber, lipids, vitamin E, and various minerals. Its commercialization and transport are generally done with the fruit in pulp form, which gives it greater stability as it allows freezing without compromising its characteristics. Petition 870250012114, dated 02 / 14 / 2025, page 12 / 60 5 / 39 sensory. Products obtained from açaí are classified based on the amount of water used in their processing and the solids content, with açaí pulp being obtained without the addition of water or filtration; thick açaí being obtained with the addition of water (total solids above 14%) and subsequent filtration; medium açaí, with the addition of water (total solids between 11 and 14%) and subsequent filtration; and thin açaí, with the addition of water (total solids between 8 and 11%) and subsequent filtration.

[0016] Its pulp is rich in bioactive compounds, containing antimicrobial, antioxidant, anti-inflammatory, analgesic, and cardioprotective action. Anthocyanins are the main compounds present in açaí pulp, and they give it its purple color. These characteristics related to anthocyanins have increased the industry's interest in its use and commercialization.

[0017] Documents EP1981513B1 and US8048456B2 address the use of Euterpe oleracea extracts in cosmetic formulations, with antioxidant and regenerative characteristics. Documents US20200390696A1 and RU2690651C2 describe a beverage composed of a fibrous layer together with Euterpe oleracea extract (in addition to other plants). Thus, it is seen that the documents presenting the use of Euterpe oleracea Mart. aim at its use as a cosmetic, medicinal or food product, but do not address its use as a dye. Its use as a dye was investigated in the studies of CIPRIANO, Paula de A. (2011), whose master's thesis “ANTHOCYANINS FROM AÇAÍ (EUTERPE OLERACEA MART.) AND JABUTICABA PEEL (MYRCIARIA JABOTICABA) IN THE FORMULATION OF ISOTONIC BEVERAGES”, which presents the use of anthocyanin extracts from mixtures of jabuticaba peel and açaí pulp in the formulation and characterization of isotonic beverages.This thesis does present the potential of açaí as a coloring agent, but only when applied directly to the formulation of beverages, and it doesn't reveal any concrete evidence. Petition 870250012114, dated 02 / 14 / 2025, page 13 / 60 6 / 39 coloring composition that can be applied to a wide variety of products.

[0018] Furthermore, another plant-based raw material with potential use as a natural pigment is beetroot (Beta vulgaris). Beetroot is used worldwide, with diverse applications ranging from culinary uses to the production of alcohol or sugar. Its tuberous crown is the most widely used part and of greatest interest to industry, being an important source of compounds called betalains. Betalains are pigments originating from plants that have gained more space in the current food market due to their natural appeal. In addition to their coloring power, betalains also exhibit important bioactive characteristics, such as reducing the size of mutant cells stimulated by nitrosamines. Beetroot juice, which combines betalains with other compounds, shows potential for prevention against vascular and heart diseases and cancer.

[0019] Its use as a pigment has been commercially explored, mainly in the manufacture of bread and meat products. For example, the studies by MEIRELES, et al. (2020), entitled EVALUATION OF THE COLORING AND ANTIOXIDANT POTENTIAL OF BETALAINS (BETA VULGARIS, L.) IN CHICKEN MORTADELLA, published in Research, Society and Development, investigate the use of betalains as a natural coloring agent and antioxidant in mortadella, and patent JP7241802B2 also suggests the use of beetroot extract as a food coloring for meats, so it is seen that the use of betalains as a coloring agent has restricted application.

[0020] As mentioned earlier, natural dyes often have problems related to color stability and microbiological stability, which are relevant factors for their application. Therefore, it is seen that no prior art document presents... Petition 870250012114, dated 02 / 14 / 2025, page 14 / 60 7 / 39 a means of obtaining plant-based colorants that have high stability and wide application in various foods.

[0021] Thus, the present invention proposes a process for preparing a red dye of vegetable origin, comprising obtaining and mixing hydroalcoholic extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris, and said dye composition comprising hydroalcoholic extracts, so that the present invention represents a vegan, biodegradable alternative with reduced allergenicity, compared to existing dyes. BRIEF DESCRIPTION OF THE INVENTION:

[0022] The present invention describes processes for preparing a dye of natural origin comprising hydroalcoholic extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris; and dye compositions comprising said extracts.

[0023] The present invention, by combining extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris, provides an increase in the stability of the dye composition through the phenomenon of co-pigmentation. Furthermore, by combining different proportions of the extracts, it allows the creation of a wide range of possible colors.

[0024] Thus, in one embodiment of the present invention, processes are disclosed for the preparation of a dye of vegetable origin comprising obtaining extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris and subsequent mixing of these extracts, wherein the obtaining of the extracts comprises contact of the plant material with the extraction solvent, mechanical agitation, and cold pressing. In one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises the steps: a) pre-preparation of the plant raw material, wherein the plant raw material is selected from F. chica, E. oleracea and / or Beta vulgaris; b) preparation of the extraction solvent comprising mixing of Petition 870250012114, dated 02 / 14 / 2025, page 15 / 60 8 / 39 citric acid in 70% ethanol, at a concentration of about 0.01 to 10.0%, preferably at a concentration of about 0.1%; c) contact the solvent with the plant raw material in a plant:solvent ratio of about 1:1 to about 1:10 (w / v), preferably about 1:3 (w / v); d) mechanical stirring for 30 to 180 min, preferably for about 60 min; e) cold pressing; f) segregation and vacuum filtration of the liquid material obtained; g) addition of new solvent to the residual solid material; h) repeat steps c) to g) two to three times; i) drying of the filtered material obtained in a rotary evaporator with reduced pressure; wherein the material obtained is a paste with a water content between about 20% and approximately 50%. In a preferred embodiment of the present invention, the material obtained is a paste with a water content of approximately 50% (w / w). In one embodiment of the present invention, the pre-preparation of crajiru comprises drying the crajiru leaves at a temperature of approximately 30°C to approximately 60°C, preferably approximately 40°C; and grinding until a granulation of less than approximately 40 mesh is obtained. In a preferred embodiment of the invention, the grinding is done in a knife mill. In one embodiment of the present invention, the pre-preparation of E. oleracea comprises obtaining the freeze-dried coarse pulp by freeze-drying the coarse pulp for approximately 18h to 48h, preferably 24h, at temperatures between -10°C and -60°C, preferably -45°C. In one embodiment of the present invention, the pre-preparation of Beta vulgaris comprises the use of the peeled and sectioned tuber neck.In one embodiment of the present invention, the cold pressing step of the mixture occurs in a screw-type press. In another embodiment of the present invention, drying occurs in a rotary evaporator with reduced pressure at a temperature of about 40°C to about 50°C. In a preferred embodiment of the present invention, drying occurs in a rotary evaporator with rotating pressure. Petition 870250012114, dated 02 / 14 / 2025, page 16 / 60 9 / 39 temperature of about 40°C. In one embodiment of the present invention, the hydroalcoholic extracts are obtained separately or in combination. In one embodiment of the present invention, the process further comprises the step of mixing the extracts in paste form. In one embodiment of the present invention, the step of mixing the extracts comprises mixing the extracts of E. oleracea, B. vulgaris, F. Chica in proportions of about 1:1:1 to about 40:5:1. In one embodiment of the present invention, the step of mixing the extracts comprises mixing the extracts of E. oleracea, B. vulgaris, F. Chica in proportions of about 1:1:1 to about 5:40:1. In one embodiment of the present invention, the step of mixing the extracts comprises mixing the extracts of E. oleracea, B. vulgaris, F. Chica in proportions of approximately 1:1:1 to about 1:5:40. In one embodiment of the present invention, the mixing step of the extracts comprises mixing the extracts of E. oleracea, B. vulgaris, F. Chica in proportions of about 1:1:1 to about 40:1:5. In one embodiment of the present invention, the dye obtained by the process has a red, pink, or purple color.

[0025] In one embodiment of the present invention, compositions comprising hydroalcoholic extracts of Fridericia chica, Euterpe oleracea, and Beta vulgaris are disclosed. In a further embodiment of the present invention, the composition comprises hydroalcoholic extracts of Fridericia chica, Euterpe oleracea, and Beta vulgaris in proportions of approximately 1:1:1 to about 40:5:1, from about 1:1:1 to about 5:40:1, from about 1:1:1 to about 1:5:40 and from about 1:1:1 to about 40:1:5. In a further embodiment, the composition comprising hydroalcoholic extracts additionally comprises co-pigments from other plant sources. In a further embodiment, the composition comprising hydroalcoholic extracts has a moisture content between about 20 and about 60%, preferably about 35%. In a Petition 870250012114, dated 02 / 14 / 2025, p. 17 / 60 In an additional embodiment, the composition comprising hydroalcoholic extracts has a paste-like form. In a further embodiment, the composition comprising hydroalcoholic extracts has an acidic pH. In a preferred embodiment, the composition comprising hydroalcoholic extracts has a pH less than 6. In a preferred embodiment, the composition comprising hydroalcoholic extracts has a pH less than 5. In a further embodiment, the composition comprising hydroalcoholic extracts is a dye composition. In a further embodiment, the composition comprising hydroalcoholic extracts is a red, orange, pink, or purple dye. In a further embodiment, the composition comprising hydroalcoholic extracts exhibits high color stability. In a further embodiment of the present invention, the composition comprising hydroalcoholic extracts exhibits good microbiological stability.In a further embodiment of the present invention, the composition comprising hydroalcoholic extracts is a dye for food, cosmetics or textiles. In another embodiment of the present invention, the composition comprising hydroalcoholic extracts is a vegan dye. In a further embodiment, the composition comprising hydroalcoholic extracts is obtained by the process for preparing a dye of vegetable origin of the present invention.

[0026] In a further embodiment of the present invention, uses of the composition comprising the hydroalcoholic extracts as a dye are disclosed. In a further embodiment of the invention, the use is in the manufacture of food, nutraceuticals or cosmetics. In a further embodiment of the invention, the use is for coloring fabrics. BRIEF DESCRIPTION OF THE FIGURES:

[0027] Figure 1 shows the UV-vis spectrometry graph of the dyes in isolation and in Petition 870250012114, dated 02 / 14 / 2025, page 18 / 60 11 / 39 set, compared to synthetic dyes, in which the wavelengths for the hydroalcoholic extracts and the dye composition of the present invention are presented, in comparison with synthetic dyes. Sunset Yellow dye; AZO dye; Red 40 dye; Açaí: açaí extract; Beetroot: beetroot extract; F. chica: crajiru extract; ABC dye: dye composition comprising the mixture of hydroalcoholic extracts of açaí, beetroot and crajiru.

[0028] Figure 2 shows the individual extracts of açaí (A), beetroot (B) and crajiru (C); as well as the coloring composition comprising the mixture of the extracts (ABC).

[0029] Figure 3 shows the color compositions (ratio 40:5:1 - E. oleracea: B. vulgaris: F. chica) diluted to 1%, 5% and 10% (w / v) in 70% (v / v) ethanol. DETAILED DESCRIPTION OF THE INVENTION:

[0030] The present invention relates to processes for preparing a dye of natural origin comprising hydroalcoholic extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris; and dye compositions comprising said extracts.

[0031] The processes for preparing natural dyes of the present invention represent an alternative to meet commercial needs for natural products, especially those of plant origin, since the vegan market lacks natural plant-based dyes that exhibit good color stability and microbiological stability.

[0032] The present invention, by combining extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris, provides an increase in the stability of the dye composition through the phenomenon of copigmentation. Furthermore, by Petition 870250012114, dated 02 / 14 / 2025, p. 19 / 60 Combining different proportions of the extracts (12 / 39) allows for the creation of a wide range of possible colors.

[0033] The present invention utilizes the combination of solvent use with the cold pressing step to obtain larger quantities of compounds of interest, thus promoting a higher yield.

[0034] Thus, in one embodiment of the present invention, processes are disclosed for the preparation of a dye of vegetable origin comprising obtaining extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris and subsequent mixing of these extracts, wherein obtaining the extracts comprises contact of the plant material with the extracting solvent, mechanical agitation, and cold pressing. In the present invention, the act of mixing refers to the act of mixing, combining, uniting, merging, integrating, or any type of joining, without limiting the scope of protection of the present invention.

[0035] For the preparation of plant-based dyes, pre-preparation steps for the raw materials are foreseen. The raw materials used are selected from F. chica, E. oleracea and / or Beta vulgaris, but other sources of natural pigments are also possible.

[0036] The pre-preparation of crajiru (F. chica) comprises drying the crajiru leaves at a temperature of about 30°C to about 60°C, preferably about 40°C. The drying of the leaves can be carried out in an oven, a forced-air oven, sun drying, dehydrators, desiccators or by microwave, without limiting the scope of protection of the invention.

[0037] Next, the dried leaves are crushed until a smaller granulation of about 18 mesh to 100 mesh is obtained, preferably less than about 40 mesh.

[0038] Furthermore, in the present invention, the term grinding means the act of grinding, milling, crushing, Petition 870250012114, dated 02 / 14 / 2025, page 20 / 60 13 / 39 fragmentation, or any type of comminution, without limiting the scope of protection of the present invention.

[0039] There are various methods and equipment for grinding, such as knife mills, ball mills, hammer mills, roller mills, screw mills, which should not limit the scope of protection of the present invention.

[0040] Grinding the leaves is important to allow greater solvent reach and increase extraction yield. By reducing particle size, grinding exposes the pigment, facilitating contact between the solvent and the plant, allowing for more efficient extraction.

[0041] Regarding açaí extracts, one can choose to use its fresh thick pulp, or alternatively, the pre-preparation of E. oleracea, which involves obtaining freeze-dried thick pulp to facilitate the handling of the extract and reduce the volume to be dried in a rotary evaporator. Freeze-drying facilitates handling and allows the use of less solvent for extraction, as it avoids the step of calculating the interference of water present in the fresh material for extraction. This reduces the volume of extract obtained, but does not necessarily reduce its yield (with fresh pulp, a larger volume is obtained, but of a more diluted extract).

[0042] Obtaining freeze-dried coarse pulp involves freeze-drying the coarse pulp for about 18 h to 48 h, preferably 24 h, at temperatures between -10 °C and -60 °C, preferably -45 °C. Freeze-dried coarse pulp can also be obtained commercially through third parties. In addition, the present invention also provides for the use of coarse pulp without being freeze-dried.

[0043] In turn, with regard to beetroot pigment, the pre-preparation of Beta vulgaris involves obtaining the tuberous neck, peeled and sectioned into 4 to 20 sections, preferably 10 sections. The tuberous neck can be used fresh (approximate water content between 50% and 98%), Petition 870250012114, dated 02 / 14 / 2025, p. 21 / 60 14 / 39 preferably approximately 87%) or freeze-dried. Freeze-drying facilitates handling and allows for the use of less solvent for extraction, as it avoids the step of calculating the interference of water present in the fresh material for extraction.

[0044] Obtaining lyophilized tuber necks involves lyophilizing the tuber necks for approximately 18 to 48 hours, preferably 24 hours, at temperatures between -10 °C and -60 °C, preferably -45 °C.

[0045] Pre-preparation of raw materials makes it possible to increase the yield of the extraction step, since it allows increasing the particle size and exposing the pigments, enabling the solvent to be used to extract more pigments of interest. Thus, in one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises a pre-preparation step of the vegetable raw material, in which the vegetable raw material is selected from F. chica, E. oleracea and / or Beta vulgaris. In one embodiment of the present invention, the pre-preparation of F. chica comprises drying the F. chica leaves at a temperature of about 30°C to about 60°C, preferably about 40°C; and grinding until a granulation of less than about 40 mesh is obtained. In a preferred embodiment of the invention, the grinding is done in a knife mill. In one embodiment of the present invention, the pre-preparation of E. oleracea comprises obtaining freeze-dried coarse pulp.In a preferred embodiment of the present invention, obtaining the freeze-dried thick pulp comprises freeze-drying the thick pulp for about 18 to 48 hours, preferably 24 hours, at temperatures between -10 °C and -60 °C, preferably -45 °C. In one embodiment of the present invention, the pre-preparation of Beta vulgaris comprises obtaining the peeled and sectioned tuber neck, fresh or freeze-dried. In another embodiment of the present invention, obtaining the freeze-dried tuber neck. Petition 870250012114, dated 02 / 14 / 2025, p. 22 / 60 15 / 39 involves freeze-drying the tuber neck for approximately 18 to 48 hours, preferably 24 hours, at temperatures between -10°C and -60°C, preferably -45°C.

[0046] The choice of extraction solvent used for extracting pigments from plants is important to ensure that the solvent is able to solubilize and extract the compounds of interest. A good solvent allows for the selection of desired compounds, avoiding the extraction of impurities. The solvent used must also be non-toxic and compatible with the material to be extracted. Furthermore, the solvent should have characteristics that do not damage the compounds of interest and, ideally, have a low boiling point, allowing for easy removal from the mixture (concentrating the compounds of interest). Thus, the present invention also provides a step for preparing the extraction solvent. The extraction solvent comprises a mixture of citric acid in 70% ethanol, where the citric acid is at a concentration of 0.01% to 10.0%, preferably at a concentration of about 0.1%. The use of an acidic solvent is important because of the affinity of the compounds of interest for an acidic medium. When found in their ionized form (generally in an acidic medium), anthocyanins become more soluble in solvents of greater polarity, such as water and ethanol.When in their molecular form (generally in neutral and basic media), they are more soluble in highly polar solvents such as hexane and chloroform. Furthermore, anthocyanins are acidic molecules, and their color is closely related to the pH of the medium in which they are found. In basic pH, they exhibit a yellowish or bluish color (and may even irreversibly change color in very basic pH). In acidic pH, they retain their orange, red, or purple color. Petition 870250012114, dated 02 / 14 / 2025, page 23 / 60 16 / 39

[0047] Thus, in one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises a step of preparing the extraction solvent comprising mixing citric acid in 70% ethanol, at a concentration of 0.01% to 10.0%, preferably at a concentration of about 0.1%.

[0048] The formulated extraction solvent can be used to obtain extracts from different raw materials.

[0049] The present process also provides for contact between the raw material and the solvent, in which the plant and the solvent are contacted in a ratio of about 1:1 to about 1:10 (w / v), preferably about 1:3 (w / v). The contact must occur for a sufficient time to allow the extraction of the compounds. In addition, the extraction can be improved when subjected to agitation. The mechanical agitation and the contact time between plant and solvent can be from about 30 to 180 min, preferably for about 60 min.

[0050] Thus, in one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises a step of contacting the solvent with the vegetable raw material in a plant:solvent ratio of about 1:1 to about 1:10 (w / v), preferably about 1:3 (w / v). In one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises a step of mechanical agitation for 30 to 180 min, preferably for about 60 min.

[0051] The present invention provides for cold pressing of the plant and solvent mixture in order to intensify the extraction of the compounds. Cold pressing promotes closer contact between the solvent and the plant material, which aids in the extraction of the compounds. Furthermore, the process is also able to maintain the integrity of the compounds, since it does not use heat, which causes degradation of the pigments of interest in the extract. Pressing aids in the extraction process by Petition 870250012114, dated 02 / 14 / 2025, page 24 / 60 17 / 39 not only allows the extraction of dyes soluble in the hydroalcoholic solvent, but also allows the extraction of insoluble (or partially insoluble) fractions that may be carried away by the physical force exerted by pressing. Cold pressing can occur in hydraulic presses, mechanical presses, and screw presses, without limiting the invention. Thus, in one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises a cold pressing step. In one embodiment of the present invention, the cold pressing step of the mixture occurs in a screw press.

[0052] The process for preparing plant-based dyes also includes a segregation and vacuum filtration step of the liquid material obtained, so that the residual solid part can undergo solvent extraction and cold pressing again, to increase the amount of extracted compounds. Even after solvent extraction, some of the pigments are still retained in the plant material, so a new extraction is desirable. Considering the saturation of the extracting solvent, the residual solid material is extracted with a new, unsaturated solvent, in order to intensify the obtaining of water-soluble extracts. Thus, in one embodiment of the present invention, the process for preparing plant-based dyes comprises a segregation and vacuum filtration step of the liquid material obtained. In another embodiment of the present invention, the process for preparing plant-based dyes comprises a step of adding a new solvent to the residual solid material.

[0053] In order to increase the process yield, the present invention provides for the repetition of steps to extract more compounds. Thus, in one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises repeating the steps of contacting the plant material with the solvent, agitation Petition 870250012114, dated 02 / 14 / 2025, page 25 / 60 18 / 39 mechanical, cold pressing, segregation and filtration of the material and addition of new solvent two to three times.

[0054] After obtaining hydroalcoholic extracts of F. chica, E. oleracea, or Beta vulgaris, the extracts are dried to obtain a paste-like form. The inventors surprisingly found that the coloring composition of the present invention, by presenting the extracts dissolved in a hydroalcoholic medium, allows its incorporation into various applications. Thus, the extracts obtained by the process of the present invention are dried until a paste with a water content of about 20% to about 50% is obtained, preferably about 50%. Drying can occur in dryers that allow relatively low temperatures (approximately 50 °C), in order to preserve the characteristics of the compounds of interest, preferably where drying is carried out in a rotary evaporator with reduced pressure, without limiting the scope of protection of the present invention. Furthermore, drying can occur at temperatures of about 40°C to about 50°C, preferably about 40°C.Thus, in one embodiment of the present invention, the process for preparing a dye of vegetable origin comprises a drying step of the filtered material obtained in a rotary evaporator with reduced pressure; wherein the material obtained is a paste with a water content between about 20% and about 50%. In a preferred embodiment of the present invention, the material obtained is a paste with a water content of about 50% (m / m). In one embodiment of the present invention, the drying occurs in a rotary evaporator with reduced pressure at a temperature of about 40°C to about 50°C. In a preferred embodiment of the present invention, the drying occurs in a rotary evaporator with rotary pressure at a temperature of about 40°C.

[0055] The extracts obtained have a paste-like texture and are mixed to prepare the vegetable-based dye. In the present invention, the act of mixing refers to the act of Petition 870250012114, dated 02 / 14 / 2025, page 26 / 60 19 / 39 to mix, combine, unite, merge, integrate, or any type of joining, without limiting the scope of protection of the present invention. The combination of extracts allows the resulting dye composition to have greater stability due to the co-pigmenting effect between the mixture of three different extracts. The co-pigmentation effect is achieved by using two extracts containing mainly anthocyanins and one extract containing alkaloids.

[0056] Thus, in order to obtain a wide range of available colors, the extracts of E. oleracea, B. vulgaris, and F. Chica can be combined in different proportions. The present invention provides that the extracts of E. oleracea, B. vulgaris, and F. Chica can be in proportions of about 1:1:1 to about 40:5:1; proportions of about 1:1:1 to about 5:40:1; proportions of about 1:1:1 to about 1:5:40; or proportions of about 1:1:1 to about 40:1:5. Thus, the dyes obtained by the process of the present invention represent a system of at least 3 components, which has more complex behavior than vegetable dyes from only one vegetable source. Thus, in one embodiment of the present invention, the process further comprises the step of mixing the extracts in paste form. In one embodiment of the present invention, the mixing step of the extracts comprises mixing the extracts of E. oleracea, B. vulgaris, and F. chica in proportions of about 1:1:1 to about 40:5:1.In one embodiment of the present invention, the extract mixing step comprises mixing the extracts of... E. oleracea, B. vulgaris, F. Chica in proportions of approximately 1:1:1 to approximately 5:40:1. In one embodiment of the present invention, the extract mixing step comprises mixing the extracts of E. oleracea, B. vulgaris, and F. chica in proportions of approximately 1:1:1 to approximately 1:5:40. In one embodiment of the present invention, the extract mixing step comprises mixing the extracts of E. oleracea, Petition 870250012114, dated 02 / 14 / 2025, p. 27 / 60 20 / 39 B. vulgaris, F. Chica in proportions of about 1:1:1 to about 40:1:5.

[0057] By combining extracts of E. oleracea, B. vulgaris and F. Chica, the process of the present invention promotes stabilization of the dye through chemical means, by the co-pigmentation action of the molecules present in the other extracts.

[0058] The co-pigmenting potential between the mixture of three different extracts used in the present invention is achieved by using two extracts containing mainly anthocyanins and one extract containing alkaloids.

[0059] Due to the co-pigmentation effect, the dye preparation process of the present invention allows for a wider range of accessible colors, such that the dyes obtained exhibit colors ranging from red, orange, pink, and purple, depending on the proportion used between the extracts. Furthermore, due to the co-pigmentation effect, the dye preparation process of the present invention also allows the produced dyes to have greater stability.

[0060] The color of the dyes varies depending on the proportion of each extract used, with the addition of more crajiru extract resulting in orange and red colors, the addition of more beetroot extract resulting in pink and purple colors, and the addition of açaí extract resulting in purple and brownish colors.

[0061] Thus, in one embodiment of the present invention, the dye obtained by the present process has a red, orange, pink or purple color.

[0062] The process of the present invention allows the preparation of compositions comprising hydroalcoholic extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris, which, when combined in different proportions, allows the obtaining of a wide range of colors. The extracts of Petition 870250012114, dated 02 / 14 / 2025, p. 28 / 60 21 / 39 Fridericia chica, Euterpe oleracea, and Beta vulgaris can be combined in ratios of approximately 1:1:1 to approximately 40:5:1, approximately 1:1:1 to approximately 5:40:1, approximately 1:1:1 to approximately 1:5:40, and approximately 1:1:1 to approximately 40:1:5. The ratio between the extracts will vary depending on the desired color. In addition to extracts of Fridericia chica, Euterpe oleracea, and Beta vulgaris, the coloring composition optionally includes co-pigments from other plant sources. Copigments can provide the anthocyanins cyanidin 3-glucoside and cyanidin 3-rutinoside, alkaloids (betalaine), carotenes, flavonoids, rutin, and ferulic acid, whose purpose is related to maintaining the final color of the product against humidity, temperature, or light exposure. Thus, in one embodiment of the present invention, compositions comprising hydroalcoholic extracts of Fridericia chica, Euterpe oleracea, and Beta vulgaris are disclosed.In a further embodiment of the present invention, the composition comprises hydroalcoholic extracts of Fridericia chica, Euterpe oleracea and Beta vulgaris in proportions of about 1:1:1 to about 40:5:1, of about. 1:1:1 to about 5:40:1, about 1:1:1 to about 1:5:40 and about 1:1:1 to about 40:1:5. In a further embodiment, the composition comprising hydroalcoholic extracts additionally comprises co-pigments from other plant sources. The co-pigmenting potential between the mixture of three different extracts used in the present invention is achieved by using two extracts containing mainly anthocyanins and one extract containing alkaloids.

[0063] The dyes generally available on the market are in powder form, which can hinder their application. Thus, the inventors surprisingly found that the dye of the present invention, being in paste form and comprising extracts dissolved in a medium Petition 870250012114, dated 02 / 14 / 2025, p. 29 / 60 22 / 39 hydroalcoholic, allows its incorporation into other inputs and various applications. The composition comprising hydroalcoholic extracts has a moisture content between approximately 20% and approximately 60%. The hydroalcoholic extract composition has a paste-like texture.

[0064] The hydroalcoholic extract composition can be diluted or used in its concentrated form, depending on the desired application. In one embodiment of the present invention, the hydroalcoholic extract composition can be diluted in 70% ethanol, to concentrations of about 1% to about 10%. When diluted, the kinematic viscosity of the diluted hydroalcoholic extract composition (from 1% to 10%) can vary from about 0.85 to about 1.10 mm² / s. When undiluted, that is, in its obtained form, the hydroalcoholic extract composition exhibits a kinematic viscosity in the range of about 2.3 mm² / s to about 2.9 mm² / s.

[0065] Thus, in a further embodiment, the composition comprising hydroalcoholic extracts has a moisture content between about 20% and about 60%. In a preferred embodiment, the composition comprising hydroalcoholic extracts has a moisture content of about 35%. In a further embodiment, the composition comprising hydroalcoholic extracts has a paste-like form. In a further embodiment, the hydroalcoholic extract composition can be diluted or used in its concentrated form.

[0066] In a further embodiment, the composition comprising hydroalcoholic extracts has a kinematic viscosity of approximately 0.9052 mm2 / s for a 70% (v / v) ethanol solution containing 1% (m / v) of a mixture of the extracts in a 40:5:1 ratio (E. oleracea: B. vulgaris: F. (chica). In a further embodiment, the composition comprising hydroalcoholic extracts has a kinematic viscosity of approximately 0.9605 mm² / s for a solution. Petition 870250012114, dated 02 / 14 / 2025, p. 30 / 60 23 / 39 of 70% (v / v) ethanol containing 5% (w / v) of a mixture of the extracts in a 40:5:1 ratio (E. oleracea: B. vulgaris: F. chica). In a further embodiment, the composition comprising hydroalcoholic extracts has a kinematic viscosity of approximately 1.0605 mm² / s for a 70% (v / v) ethanol solution containing 10% (w / v) of a mixture of the extracts in a 40:5:1 ratio (E. oleracea: B. vulgaris: F. chica). Thus, in one embodiment of the present invention, the composition comprising hydroalcoholic extracts has a kinematic viscosity in the range of 2.9 mm² / s to 0.9 mm² / s, preferably, wherein it has a kinematic viscosity in the range of about 2.8119 mm² / s to about 0.9052 mm² / s.

[0067] In addition to the co-pigmentation effect, the colors of the composition comprising hydroalcoholic extracts are stabilized due to the pH of the composition, which, being acidic, allows the color to remain stable for longer. Thus, in a further embodiment, the composition comprising hydroalcoholic extracts has an acidic pH. In a preferred embodiment, the composition comprising hydroalcoholic extracts has a pH lower than about 6. In a preferred embodiment, the composition comprising hydroalcoholic extracts has a pH lower than about 5.

[0068] The co-pigmenting potential between the mixture of three different extracts used in the present invention is achieved by using two extracts containing mainly anthocyanins and one extract containing alkaloids.

[0069] The composition comprising water-soluble extracts of the present invention is a coloring composition that possesses high color stability and good microbiological stability. Furthermore, the coloring composition of the present invention has various applications, such as in the preparation of food, nutraceuticals, cosmetics, or in dyeing fabrics. The present invention meets the Petition 870250012114, dated 02 / 14 / 2025, page 31 / 60 24 / 39 contemporary trends in natural, vegan, biodegradable, low-allergenicity, and cruelty-free dyes.

[0070] Thus, in a further embodiment, the composition comprising hydroalcoholic extracts is a dye composition, with colors selected from red, orange, pink or purple. In a further embodiment, the composition comprising hydroalcoholic extracts exhibits high color stability. The present invention offers a technological advantage by providing dyes with high color and microbiological stability, factors that are relevant for plant-derived dyes and their applications.

[0071] In a further embodiment of the present invention, the composition comprising hydroalcoholic extracts exhibits good microbiological stability. In a further embodiment of the present invention, the composition comprising hydroalcoholic extracts is a dye for food, cosmetics or textiles. In one embodiment of the present invention, the composition comprising hydroalcoholic extracts is a vegan dye. In a further embodiment, the composition comprising hydroalcoholic extracts is obtained by the process for preparing a dye of vegetable origin of the present invention.

[0072] The coloring composition of the present invention is capable of providing the anthocyanins cyanidin 3-glucoside and cyanidin 3-rutinoside, alkaloids (betalaine), carotenes, flavonoids, rutin and ferulic acid.

[0073] The dyes obtained in the present invention have wide application in various products in the food, cosmetic or textile industries, and it is also important to emphasize the fact that they are dissolved in a hydroalcoholic medium, which facilitates their incorporation into other ingredients.

[0074] Thus, the present invention discloses the use of the composition comprising hydroalcoholic extracts as colorants, which can be used in the manufacture of Petition 870250012114, dated 02 / 14 / 2025, page 32 / 60 25 / 39 foods, nutraceuticals or cosmetics. The dyes of the present invention can also be used to dye fabric.

[0075] Thus, in a further embodiment of the present invention, uses of the composition comprising the hydroalcoholic extracts as a dye are disclosed. In a further embodiment of the invention, the use is in the manufacture of food, nutraceuticals or cosmetics. In a further embodiment of the invention, the use is for coloring fabrics.

[0076] The present invention provides natural colorants and contributes to the establishment of an industry with reduced environmental damage, respecting current consumer demand.

[0077] The invention can also be further described by the following non-limiting examples. The following examples detail and explain better the performance of the inventive processes. Those skilled in the art will recognize many variations that are within the spirit of the invention and the scope of the claims. EXAMPLES OF IMPLEMENTATION:

[0078] Example 1: In an embodiment of the present invention, dye compositions were prepared according to the method of the present invention. Initially, hydroalcoholic extracts were obtained from each plant raw material (crajiru, açaí and beetroot). The dye composition of the present invention was prepared from the mixture of the hydroalcoholic extracts.

[0079] For this purpose, leaves and stems of F. chica (crajiru) were collected and sun-dried for 2 days at an average temperature of approximately 40°C. The dried crajiru was ground in a knife mill to a granulation smaller than 40 mesh. For E. oleracea (açaí), its freeze-dried thick pulp was obtained, and for B. vulgaris (beetroot), its peeled and sectioned (scraped) tuberous collar was obtained in its fresh form. Then, the extraction solvent was prepared. Petition 870250012114, dated 02 / 14 / 2025, page 33 / 60 26 / 39 comprising a mixture of citric acid in 70% ethanol, at a percentage of 0.1%. This solvent is the same one used for the extraction of pigments from the three plant species.

[0080] Extracts from each plant were obtained individually. For this, the plant material and the extraction solvent were contacted in a ratio of 1:3 (w / v) for 60 min under mechanical agitation. Then, the solvent+plant mixture was placed in a screw press for cold pressing. Cold pressing is important because it promotes closer contact between the solvent and the plant material, and also further reduces the particle size of the plant material, increasing the contact surface and providing a higher extraction yield. Subsequently, the liquid material obtained was segregated and vacuum-filtered, and new solvent was added to the remaining solid part.

[0081] The addition of new solvent is important so that the solvent is not saturated with components, in order to increase the yield of the extraction process. The steps of solvent contact with the plant material, mechanical agitation, cold pressing and segregation of the material, and addition of new solvent were repeated 3 times, for better extraction of the compounds of interest.

[0082] Next, the obtained material was dried in a rotary evaporator with reduced pressure at 40°C, where the material was dried until a paste with a water content of 50% (m / v) was obtained. The extracts were obtained individually, presenting reddish tones, as shown in Figure 2.

[0083] In order to obtain the color composition of the present invention, the extracts were mixed in proportions of 40:5:1 (E. oleracea: B. vulgaris: F. chica). The color composition has a red color, also shown in Figure 2 and Figure 3. In Figure 3, they are Petition 870250012114, dated 02 / 14 / 2025, page 34 / 60 27 / 39 presented the color compositions diluted to 1%, 5% and 10% (w / v) in 70% (v / v) ethanol.

[0084] The hydroalcoholic extracts and the paste-textured coloring composition were stored in a closed container, protected from light, under refrigeration at 8 °C, until the time of testing.

[0085] The extracts of the present invention were analyzed considering the intensity of coloration, quantity of monomeric anthocyanins, pH variation, general humidity, dye darkening and microbiological stability tests. The average extraction yield of the species was 30.99% (w / w) for açaí, 49.17% (w / w) for beetroot and 23.57% (w / w) for crajiru.

[0086] Evaluation of color intensity by UV-Vis: The extracts were evaluated based on their coloration using the UV-Vis scanning technique. Different masses of the samples were weighed in a volumetric flask and their volume was completed with 70% ethanol, in order to generate a solution with sufficient concentration for reading in UV-Vis equipment. For the extracts, solutions of 2.58 mg / L of açaí extract, 0.34 mg / L of beetroot, and 0.062 mg / L of crajiru were prepared. For the color composition, a solution comprising 2.58 mg / L of açaí extract, 0.34 mg / L of beetroot, and 0.062 mg / L of crajiru (i.e., in a 40:5:1 ratio) was prepared. The blank test (control) was performed only with 70% ethanol. The mixture was homogenized and evaluated by UV-vis spectroscopy using an Agilent® 8453 UV-vis spectrophotometer, with 70% ethanol as a blank.Its coloring power was evaluated by comparing the extract to the dyes Red 40, Sunset Yellow, and AZO, used as standards for red, orange, and pink coloring, respectively, focusing on the spectra from 460 to 600 nm (red region). The coloring strength was compared using the mass values ​​of the dye used in... Petition 870250012114, dated 02 / 14 / 2025, page 35 / 60 28 / 39 in relation to the intensity of peaks obtained.

[0087] For açaí, absorption spectra were read from 400 to 600 nm, with a peak preferentially at approximately 525 nm. In turn, for beetroot extracts, absorption spectra were read from 450 to 600 nm, with peaks preferentially between 480 and 540 nm. For crajiru extracts, absorption spectra were read from 380 to 550 nm, with a peak preferentially at 490 nm. For the dye composition, absorption spectra were read from 380 to 600 nm for the set of extracts, with a peak preferentially at 485 nm.

[0088] When comparing the plant extracts and the coloring composition of the present invention with synthetic dyes, it can be observed that the wavelength of greatest similarity among all the dyes used is at 485 nm, making them mostly similar to sunset yellow, with a more orange hue. It is also possible to observe a slight increase in the 540 nm region for beetroot and açaí, giving the mixture a more reddish hue, as can be seen in Figure 1.

[0089] Figure 1 shows the wavelengths for the hydroalcoholic extracts and the dye composition of the present invention, in comparison with synthetic dyes, wherein Sunset Yellow dye; AZO dye; Red 40 dye; Açaí: açaí extract; Beetroot: beetroot extract; F. Chica: crajiru extract; ABC dye: coloring composition comprising a mixture of hydroalcoholic extracts of açaí, beetroot and crajiru (in a 40:5:1 ratio).

[0090] Regarding coloring power, a difference in coloring power was observed between the dyes of the present invention and synthetic dyes. In order to achieve the same coloring power as ~0.016 mg / mL of synthetic dyes, a concentration of 2.585, 0.340 and 0.062 mg / mL of açaí, beetroot and crajiru extracts, respectively, was required. Petition 870250012114, dated 02 / 14 / 2025, page 36 / 60 29 / 39 The synthetic dyes used in the test had their concentration reduced so that they did not exceed absorbance values ​​>1. For a representative base, 161 times more açaí extract, 21 times more beetroot extract, and 4 times more crajiru extract would be needed to achieve the coloration obtained with the synthetic dyes. All values ​​were calculated considering the extracts without water.

[0091] Stability screening test: To screen the stability of the dye samples, a stability screening test was performed by weighing the paste extracts separately and mixed in the dye composition of the invention, packaged in amber bottles. The stability conditions were as follows, based on the cosmetic stability manual: 24 ha 50 ± 2 °C and 24 ha -5 ± 2 °C for 6 days. The samples were analyzed at 0, 2, and 6 days. The tests performed were UV-vis staining, total monomeric anthocyanins, moisture, and pH.

[0092] Quantification of total monomeric anthocyanins: Both crajiru and açaí, present in the dye composition, are rich in anthocyanins, so the quantification of anthocyanins represents a relevant test to evaluate their degradation over time, to verify the stability of the dye composition. For this, the amount of anthocyanins present in the açaí extract, in the crajiru extract, and in the dye composition comprising the three extracts was evaluated at the time of extraction, and in comparison to storage times, highlighting losses in these components important for their coloration.

[0093] 0.8 mg were weighed into a 50 mL volumetric flask, and the volume was completed using two different buffer solutions: potassium chloride at pH 1.0 and another with sodium acetate buffer at pH 4.5. The absorbances were determined using the maximum values ​​of Petition 870250012114, dated 02 / 14 / 2025, page 37 / 60 30 / 39 absorbance of each sample observed in spectra >400 nm, and the absorbance at 700 nm. After acquiring the results, Equation 1 was applied to determine the amount of monomeric anthocyanins in the sample, with the unit of measurement expressed in cyanidin-3-glucoside equivalent in mg / L”. (A * PM * FD * 103) / £ * 1 [Equation 1] In what A: (Amax — A≈00nm) at pH 1.0 — (Amax — A≈00nm) at pH 4.5 PM: Molecular weight of cyanidin-3-glucoside = 449.2 g / mol FD: Dilution factor 103: conversion factor from g to mg £: 26900, molar coefficient, for cyanidin-3-glucoside 1: Optical path traveled (spectrophotometer cuvette)

[0094] This test revealed the presence of a cyanidin-3-glucoside equivalent of 17.16 mg / L for açaí, 6653.44 mg / L for crajiru, and 92.16 mg / L for a mixture of the three extracts. The amount of mole equivalent for beetroot did not yield valid results, as it presents a very low quantity of anthocyanins. These values ​​are summarized in Table 1: Table 1: Anthocyanins analyzed at baseline (T0), after 2 days (T2), and after 6 days (T6). Sample Signal at 485nm (A) Monomeric anthocyanins (g / mol) T0 T2 T6 T0 T2 T6 Açaí 0.304a 0. 279b 0.260c 17.2a 9.1b - Beetroot 0.322a 0. 174c 0.250b - - - Crajiru 0.264a 0.182b 0.185b 6653.4a 4447.8b 609.3c ABC Dye 0.800a 0.587b 0.498c 92.2a 49.7b 0.1c ABC dye: composition comprising açaí extracts, Petition 870250012114, dated 02 / 14 / 2025, pp. 38 / 60 31 / 39 beetroot and crajiru. Values ​​with the same letter in the same row for each of the evaluated parameters mean no statistical difference according to Tukey's test.

[0095] Note that the difference in the amount of monomeric anthocyanins is more than 300 times between the açaí extract and the crajiru extract. When comparing the extracts individually with the combination of the three extracts (in a 40:5:1 ratio), values ​​of 92 mg / mL equivalent in monomeric anthocyanins are observed, demonstrating that, even at concentrations 40 times lower than açaí and 5 times lower than beetroot, the influence of crajiru on the contribution of anthocyanins is sufficient for an increase of at least 5 times the total, when compared to the lowest value obtained (açaí extract).This is due to several factors that can reduce the percentage of anthocyanins present in açaí as a source matrix (lyophilized powder), storage time before processing (crajiru was extracted 2 days after its drying process, while açaí was stored for an unknown time) and exposure to light before processing, since, when compared to the literature, the equivalent value of monomeric anthocyanins for açaí is about 50 mg / L.

[0096] pH variation: The prepared extracts were evaluated for their average pH by weighing the extract on an analytical balance in order to obtain a 1% w / v solution dissolved in ultrapure H2O. A Micronal B474 pH meter, previously calibrated at pH 4.0 and 7.0, was used. This evaluation was performed at the initial time and during the times determined for the stability analysis.

[0097] Through analysis, pH values ​​of 4.3 were obtained for açaí, 5.8 for beetroot, and 5.0 for crajiru, demonstrating a tendency towards acidification for açaí and crajiru, and a neutral behavior for the beetroot extract. The results are summarized in Table 2. Petition 870250012114, dated 02 / 14 / 2025, pp. 39 / 60 32 / 39 Table 2: pH analyzed in the stability test at baseline (T0), after 2 days (T2), and after 6 days (T6). Sample pH T0 T2 T6 Açaí 4.30a 4.25a 4.30a Beetroot 5.78a 5.91a 5.91a Crajiru 5.05a 5.00a 5.05a ABC Dye 4.47a 4.36b 4.35b ABC dye: composition comprising extracts of açaí, beetroot, and crajiru. Values ​​with the same letter on the same line for each of the evaluated parameters indicate the absence of a statistically significant difference according to Tukey's test.

[0098] This difference may be the result of the presence of more anthocyanins in the first two extracts, acidifying them, compared to the alkaloids present in beetroot, basifying them.

[0099] Tincture Moisture: Moisture was measured using the loss on drying method. Weighing filters were kept in a Precision mechanical convection oven (GCA Corporation) at 105 °C for 1 h and subsequently placed in a desiccator containing silica gel until they reached room temperature to determine their mass. A total of 50 mg of extract were placed in weighing filters of known mass and placed in an oven at 105 °C for 4 h. After this period, the flasks were transferred to the desiccator until they reached room temperature, and the masses of the dried extracts were recorded. The difference between the final and initial mass indicates the mass of water lost by the extract, allowing the calculation of moisture. The results of the moisture test are presented in Table 3. Table 3: Moisture analyzed in the stability test at initial time (T0), after 2 days (T2) and after 6 days (T6). Sample Moisture (%) Petition 870250012114, dated 02 / 14 / 2025, pp. 40 / 60 33 / 39 T0 T2 T6 Açaí 48.3a 45.0c 46.3b Beetroot 32.7a 31.6a 32.8a Crajiru 32.5a 30.1b 30.1b ABC Dye 46.4a 42.1c 44.1b ABC Dye: composition comprising beetroot and crajiru extract. Values ​​with the same letter in the same line for each of the evaluated parameters mean absence of açaí, same line absence of statistically significant difference as determined by Tukey's test.

[00100] An average moisture content of 48.3% (m / m) was observed for the açaí extract, 32.7% (m / m) for the beet extract, and 32.5% (m / m) for the crajiru extract. These moisture values ​​facilitate the flow of the samples and their use as a paste dye at an industrial level. The dyes can be used in their concentrated paste form, or diluted, depending on the desired use. The differences in moisture content obtained are closely related to the time they remained in the rotary evaporator, being different for each sample due to the different levels of apparent viscosity during the drying process. For the açaí and beet extracts, the process took less time. This can be justified by the presence of more sugars in these two extracts (a fruit and a tuberous stem) when compared to crajiru (leaves and branches).

[00101] Evaluation of the viscosity of the dye composition: The dye composition of the present invention has a pasty characteristic. To determine the kinematic viscosity of the developed dye composition, the viscosity measurement was performed using an Ostwald / Cannon-Fenske viscometer. This methodology indicates the viscosity of a material through the speed at which it moves through a glass capillary of known dimensions, at a given temperature. For this, the glassware is filled with Petition 870250012114, dated 02 / 14 / 2025, page 41 / 60 34 / 39 the material to be studied is placed in the glassware and then negative pressure is applied so that it rises to the mark on the glassware. The pressure is then released and the liquid descends due to gravity. The time it takes for the material to migrate from the first to the second mark on the glassware is measured. The viscosity of the material is then calculated using the equation shown in the manufacturer's manual (Equation 2, an equation based on Poiseuille's equation, with the necessary corrections): v = K.(t — d) [Equation 2] Where v is the kinematic viscosity, K is the glassware constant used (0.002), t is the sample flow time in seconds, and ed is the Hagenbach-Couette correction value, provided by the manufacturer (0.07).

[00102] Considering the pasty characteristic of the composition of the present invention, a 70% (v / v) ethanol solution was prepared, in which the coloring composition in a 40:5:1 ratio (açaí:beetroot:crajiru) was dissolved in this solvent at concentrations of 1%, 5%, and 10%. Dilution was necessary because, in its undiluted form, the coloring composition is too viscous to be analyzed by this method. This material was then subjected to a viscosity test, and its results are shown in Table 4: Table 4: Average kinematic viscosity of the dye composition at different concentrations, performed at 30°C. Sample Flow time (s) Viscosity (mm² / s) Ethanol 70% 457.7 0.9152 Dye at 1% 452.7 0.9052 Dye at 5% 480.3 0.9605 Dye at 10% 530.3 1.0605

[00103] The dye composition of the present invention has a pasty characteristic, so it needs to be diluted for its viscosity to be analyzed using an Ostwald / Cannon-Fenske viscometer. Through regression Petition 870250012114, dated 02 / 14 / 2025, page 42 / 60 35 / 39 linear, it is possible to achieve a kinematic viscosity value of 2.6364 mm2 / s for the 100% dye composition, that is, without dilution.

[00104] Evaluation of color originating from polymers: This parameter evaluates the degradation index of monomeric anthocyanins. Monomeric anthocyanins are bleached by bisulfite attack, generating a colorless compound. Anthocyanins polymerized with other phenolic compounds (such as tannins) do not undergo this bleaching and retain their original color. The color density is the result of the sum of the absorbances at Xvis-max and at 420 nm (brown region), and the polymeric color is determined by the same absorbance measurement. The ratio between the absorbances with and without bisulfite treatment will reflect the contribution of the polymerized anthocyanins to the overall color of the extract.

[00105] A 2.8 mL sample of extract diluted in distilled water was added to two cuvettes. 0.2 mL of bisulfite solution was added to the first cuvette, and 0.2 mL of distilled water to the second. Both were left to stand for 15 min, and after this time their absorbance was measured in Xvis-max at 420 nm and 700 nm.

[00106] The percentage of polymeric coloration (P (%)) was calculated according to Equation 3, considering the dilution factor (DF) used for sample preparation. P (%) = CP / DC x 100 [Equation 3] Where DC: This refers to the density of coloration. CP: is the polymeric color

[00107] DC is the color density (in mixture with distilled water): DC = [(A420 - A700) + (AÀvis-max — A700) x FD] [Equation 4] In what Petition 870250012114, dated 02 / 14 / 2025, page 43 / 60 36 / 39 A420 is the absorbance at a wavelength of 420 nm. A700 is the absorbance at a wavelength of 700 nm. Aλvis-max is the maximum absorbance of each dye, being 400nm for açaí, 426nm for beetroot, 485nm for crajiru, and 530nm for the 40:5:1 mixture. FD: Dilution factor.

[00108] CP is the polymeric color (solution to which bisulfite has been added): CP = [(A420 - A700) + (AÀvis-max — A700) x FD] [Equation 5] In what A420 is the absorbance at a wavelength of 420 nm. A700 is the maximum absorbance of each dye, being 400nm for açaí, 426nm for beetroot, 485nm for crajiru and 530nm for the 40:5:1 mixture. FD: Dilution factor.

[00109] The dye polymerization test evaluates the percentage of anthocyanins that were decolorized by the action of bisulfite. For the samples present, these values ​​were equivalent to 98% for açaí, 86% for beetroot, 43% for crajiru, and 100% for the mixture of the three extracts, demonstrating that açaí and beetroot have a large contribution regarding the anthocyanins polymerized in their coloration (since these are not decolorized by bisulfite).

[00110] Crajiru showed a lower contribution from polymerized anthocyanins, demonstrating that a large part of its coloration is due to monomeric anthocyanins. After the stability test, the polymeric coloration value of each sample was 100% for açaí, 99% for beetroot, 58% for crajiru, and 91% for the mixture of the three extracts.

[00111] Based on the test results, it can be observed that the pH and moisture values ​​for the isolated extracts were very little affected during this period, which indicates that the method of the present invention provides electrolytic and hygroscopic stability for the extracts. Petition 870250012114, dated 02 / 14 / 2025, pp. 44 / 60 37 / 39 hydroalcoholic.

[00112] A significant, yet still subtle, difference in pH can only be observed in the sample that consisted of the mixture of the three extracts. This alteration may be due to possible reactions between the beetroot alkaloids (of a basic nature), the solvent system used, and the anthocyanins of the other two species. Regarding moisture, the greatest variations are observed in the açaí extracts, probably because it retains the highest initial moisture values ​​among the three extracts tested. This factor likely stems from the large amount of sugars (highly hygroscopic molecules) present in this sample, since it consisted of a pulp (crushed fruits), when compared to beetroot (tuberous collar) and crajiru (leaves).

[00113] In the mixture of extracts, it was possible to observe a behavior very similar to that of the açaí extract, justified by its predominance in the mixture (40 parts açaí, compared to 5 parts beetroot and 1 part crajiru).

[00114] The color and quantity of monomeric anthocyanins underwent significant changes. Over time, a decrease in color intensity was observed, as well as in the presence of anthocyanins in the extracts, demonstrating the instability of these extracts when subjected to drastic temperature differences (from -5 °C to 50 °C), as presented in the studies by Mattiloli et al. (2020), entitled “Anthocyanins: A Comprehensive Review of Their Chemical Properties and Health Effects on Cardiovascular and Neurodegenerative Diseases”, published in Molecules, showing the thermolabile nature of anthocyanins. This factor may also be related to the polymeric coloration results, where an increase in the rate of polymeric coloration is observed in the açaí, beetroot, and crajiru samples. Petition 870250012114, dated 02 / 14 / 2025, pp. 45 / 60 38 / 39 suggesting that the anthocyanins present in the samples have been polymerized over time. In the composition comprising the mixture of extracts, a decrease in polymeric coloration is observed, suggesting that the anthocyanins are protected from polymeric attack by the compounds present in beetroot (the only extract that does not contain monomeric anthocyanins).

[00115] Another important factor in the difference in times is the consistency of the sample. While the beetroot extract and the crajiru extract maintained their viscosity preserved during the stability test, the açaí extract showed hardening. Since its moisture values ​​are statistically different when compared to the initial moment of the test (T0), this hardening can be explained by the effect of water loss in the açaí tincture which, under the action of the temperatures used here, promotes an increase in the concentration of solids in the extract, and consequently alters its texture, as explained in the studies by Zielinska and Michalska (2016), entitled “Microwave-assisted drying of blueberry (Vaccinium corymbosum L.) fruits: Drying kinetics, polyphenols, anthocyanins, antioxidant capacity, colour and texture”, published in Food Chemistry.

[00116] Microbiological stability test: In parallel with the color stability test, a microbiological stability test was conducted to verify the ability of the extracts to remain stored without generating harmful microbial growth. This test was based on the Brazilian Pharmacopoeia, 6th edition, volume 1, using the total count of mesophilic microorganisms test.

[00117] A total of 5 mg of extract were diluted in 5 mL of 0.9% NaCl solution and homogenized. This mixture was serially diluted, generating dilutions of 1:10, Petition 870250012114, dated 02 / 14 / 2025, pp. 46 / 60 39 / 39 1:100 and 1:1000, in order to check the concentration of microorganisms in a countable way. For counting colony-forming units (CFU), the surface plate counting method was adopted. Thus, 0.1 mL of the prepared samples was added to the surface of Tryptone Soy Agar (bacteria) and Sabouraud Dextrose Agar (molds and yeasts) media and spread using a loop. Drigalski. The plates were incubated at 32.5 ± 2.5 °C for three to five days (bacteria) and 22.5 ± 2.5 °C for five to seven days (molds and yeasts). After the incubation period, the number of CFU was counted, and the average was expressed in CFU per gram or mL of product. A positive control was performed in each test to check the viability of the culture media used.

[00118] None of the extracts showed a microbial load in the count, and none of the plates showed growth. This may be due to the extracts being produced with 70% ethanol, which reduces the chances of microbial contamination.

[00119] Statistical tests: The samples were evaluated and statistically compared using Statistica software, version 8. The comparison focused on samples stored on different days, in order to observe the degradation time of the extracts. The tests performed were Analysis of Variance (ANOVA) in conjunction with multiple comparisons using Tukey's test. The significance level considered was 5%, where data were considered statistically significant for p-values ​​<0.05.

[00120] Thus, the above results demonstrate that the dye preparation process of the present invention provides a dye composition with high color stability and good microbiological stability. Petition 870250012114, dated 02 / 14 / 2025, pp. 47 / 60

Claims

1 / 4 CLAIMS 1. PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN characterized by comprising the following steps: a) pre-preparation of the vegetable raw material, in which the vegetable raw material is selected from F. chica, E.a) Extraction of *Titanium oleracea* and / or *Beta vulgaris*; b) preparation of the extraction solvent comprising mixing citric acid in 70% ethanol, at a concentration of 0.01% to 10.0%, preferably at a concentration of 0.1%; c) contacting the solvent with the plant raw material in a plant:solvent ratio of 1:1 to 1:10 (w / v), preferably 1:3 (w / v); d) mechanical agitation for 30 to 180 min, preferably for 60 min; e) cold pressing; f) segregation and vacuum filtration of the liquid material obtained; g) addition of new solvent to the residual solid material; h) repeating steps c) to g) two to three times; i) drying the filtered material obtained in a rotary evaporator with reduced pressure; wherein the material obtained is a paste with a water content between 20% and 50%, preferably 50% (w / w).

2. PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN, according to claim 1, characterized by the pre-preparation of F. chica comprising: - drying the leaves of F. chica at a temperature of 30°C to 60°C, preferably 40°C; and - grinding until a granulation of less than 40 mesh is obtained.

3. PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN, according to claim 1, characterized by the pre-preparation of E. oleracea comprising obtaining freeze-dried coarse pulp, preferably by freeze-drying the coarse pulp for 18 h to 48 h, preferably 24 h, at temperatures between -10 °C and -60 °C, preferably -45 °C.

4. PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN, according to claim 1, characterized in that the pre-preparation of Beta vulgaris comprises obtaining the peeled and sectioned tuber neck, fresh or lyophilized, preferably by lyophilizing the tuber neck for 18 h to 48 h, preferably 24 h, at temperatures between -10 °C and -60 °C, preferably -45 °C.

5. PROCESS FOR PREPARING A VEGETABLE-BASED DYE, according to claims 1 to 4, characterized in that the cold pressing of the mixture takes place in a screw-type press.

6. PROCESS FOR PREPARING VEGETABLE-BASED DYE, according to claims 1 to 5, characterized by the drying taking place in a rotary evaporator with reduced pressure at a temperature of 40°C to 50°C, preferably 40°C.

7. PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN, according to claims 1 to 6, characterized in that the hydroalcoholic extracts are obtained separately or in combination.

8. PROCESS FOR PREPARING A VEGETABLE-BASED DYE, according to claims 1 to 7, characterized by further comprising the step of mixing the extracts in paste form.

9. PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN, according to any one of claims 1 to 8, characterized by the mixing step of the extracts comprising mixing the extracts of E. oleracea, B. vulgaris, F. Chica in proportions of 1:1:1 to 40:5:1; proportions of 1:1:1 to 5:40:1; proportions of 1:1:1 to 1:5:40 Petition 870250012114, dated 02 / 14 / 2025, page 49 / 60 3 / 4 or proportions of 1:1:1 to 40:1:

5.

10. PROCESS FOR PREPARING A DYE OF VEGETABLE ORIGIN, according to any one of claims 1 to 9, characterized in that the dye has a red, orange, pink or purple color.

11. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS characterized by comprising hydroalcoholic extracts of Euterpe oleracea, Beta vulgaris and Fridericia chica in the proportions of 1:1:1 to 40:5:1, 1:1:1 to 5:40:1, 1:1:1 to 1:5:40 and 1:1:1 to 40:1:5, wherein the composition is obtained by the process as defined in claims 1 to 10.

12. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to claim 11, characterized in that it further comprises copigments from other plant sources.

13. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to claims 11 to 12, characterized by the composition having a paste-like form.

14. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to any one of claims 11 to 13, characterized in that the composition has a moisture content between 20 and 60%, preferably 35%.

15. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to claim 14, characterized in that the composition has a pH of less than 6, preferably less than 5.

16. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to any one of claims 11 to 15, characterized in that it is a coloring composition.

17. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to any of claims 11 to 16, characterized by the composition Petition 870250012114, dated 02 / 14 / 2025, page 50 / 60 4 / 4 dye being a red, orange, pink or purple dye.

18. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to any one of claims 11 to 17, characterized by the composition exhibiting high color stability.

19. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to any one of claims 11 to 18, characterized by the composition exhibiting good microbiological stability.

20. COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, according to any one of claims 11 to 19, characterized by being a vegan colorant.

21. USE OF THE COMPOSITION COMPRISING HYDROALCOHOLIC EXTRACTS, as defined in any one of claims 11 to 20, characterized in that it is used as a dye.

22. USE, according to claim 21, characterized by being in the manufacture of food, nutraceuticals, cosmetics or for dyeing fabrics. Petition 870250012114, dated 02 / 14 / 2025, pp. 51 / 60

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