Cork powder, aqueous extract, methods and uses thereof

Abstract

The present disclosure relates to a cork powder and an aqueous extract thereof and to a method for obtaining said powder and said extract. Furthermore, the present application concerns compositions comprising said powder and / or said extract and the use thereof in cosmetics or medicine.

Description

DESCRIPTION CORK POWDER, AQUEOUS EXTRACT, METHODS AND USES THEREOF TECHNICAL DOMAIN

[0001] This disclosure relates to a cork powder and an aqueous extract thereof, and a method for obtaining said powder and said extract. Furthermore, the present application relates to compositions comprising said powder and / or said extract and their use in cosmetics or medicine. BACKGROUND

[0002] The cork industry uses cork as its primary raw material, used for a variety of purposes, including the production of stoppers. Cork processing produces a considerable amount of waste, including fragments, pieces, and lower-quality cork. Given that raw material waste represents a high percentage of the product, its reuse in other industries or for the production of different materials is crucial. Therefore, this waste is often used in the manufacture of agglomerates for various purposes, such as discs, covering boards, fixing and decorative panels, insoles, and more. However, approximately 25% of the initial cork is transformed into very small granules, resulting from the various stages of its processing. This fraction is known as cork dust.

[0003] Cork powder is produced at various stages of cork processing: - A: Grinding dust - dust generated in the initial phase of granulation operations, consisting of several phases (pre-grinding and drying) before cleaning and separation operations; - B: Cleaning powder - powder obtained in the next phase involving separation - cleaning the back and belly of cork and other impurities; - C: Powder from granulometric separations - powder produced in the final granulation phase, in the separation of granules by different granulometry; - D: Powder from chipboard panel finishes - Obtained from the rolling, sanding and cutting of chipboard panels; - E: Dust from the finishing of cork stoppers and cork discs - Originating from sanding, rounding, chamfering and thickness adjustment operations; - F: Dust from the finishing of natural cork stoppers - Originated from sanding, rounding and chamfering operations. (Gil, Luis & santos, J. & Florêncio, Ml (1986). Identification and characterization of various types of powder obtained in the industrial processing of cork. IPF-Cork Bulletin. 255-261.)

[0004] Cosmetics companies are currently implementing new procedures based on a balance of economics, environment, and society to offset their environmental footprint while addressing customer concerns about sustainability and the exploitation of natural resources. As technological innovation advances, the potential and appeal of using recycled materials in cosmetics is expanding. Cork byproducts are a uniquely sustainable source due to four factors: regeneration of cork bark (every 9 years), longevity of Q. suber (up to 250 years), substantial environmental impact of Q. suber (montado) ecosystems, and production of a large volume of cork powder (per product).

[0005] The use of cork powder or an extract obtained from cork powder and its application in value-added products, particularly in the cosmetics industry, would allow for the valorization of this byproduct, contributing to the sustainable management of natural resources. However, its application in a cosmetic product presents several challenges: - Odor and color: Cork powder or powder extract has its own odorous characteristics and color, impacting the sensory characteristics and sensory acceptance by consumers; - Compatibility: Cork powder or extract may not be compatible with other ingredients present in cosmetic formulations, leading to a formulation with low stability.

[0006] Furthermore, the process of obtaining cork powder or extract can lead to the degradation of bioactive compounds, losing their biological action and the benefits of their incorporation into a cosmetic composition.

[0007] These facts are described in order to illustrate the technical problem solved by the achievements of this document. GENERAL DESCRIPTION

[0008] This disclosure relates to a cork powder and an aqueous extract thereof, and a method for obtaining said powder and said extract. Furthermore, this application also relates to compositions comprising said powder and / or said extract and their use in cosmetics or medicine.

[0009] The cork powder of this disclosure is a fine, light brown, homogeneous powder that surprisingly has been shown to have the ability to synergistically increase the sun protection factor (SPF) of sunscreens containing titanium dioxide (at least a 2-fold increase), while imparting a natural color and being compatible with the skin. It presents also excellent oil absorption properties (five times greater than kaolin) as well as low moisture content and microbiological quality, and also an adequate volatile compound profile.

[0010] The cork powder of this disclosure can be included in solid, liquid and semi-solid formulations, such as peel-off purifying masks, tinted sunscreens, dry shampoos, compacts and mattifying makeup products, among others.

[0011] The method for obtaining the cork powder described herein, particularly the powder washing and particle size reduction methods, and the microbial load reduction methods, have been optimized to produce a homogeneous and stable powder with microbiological characteristics suitable for cosmetic or medical use. The method has also been shown to be reproducible and easily scalable.

[0012] This disclosure addresses the need of the cosmetics industry and other industries for sustainable ingredients by providing an ingredient of natural origin, obtained from an agro-industrial by-product, produced in relevant quantities with consistent quality and obtained through a zero-waste process.

[0013] The multifunctionality of the cork powder of the present disclosure (fragrance, color, oil absorption and SPF reinforcement) is also attractive with regard to reducing the use of additional ingredients in the same formulation, thus reducing the carbon footprint of the products where it is used.

[0014] Another aspect of this disclosure relates to a cork powder extract. Surprisingly, the inventors of this application obtained a cork powder extract with unique characteristics, comprising cork's bioactive compounds, through the development of an innovative extraction method. This allows the obtained cork powder extract to exhibit unique characteristics, comprising cork's bioactive compounds and preserving their activity. Furthermore, the cork powder extract exhibits appropriate sensory characteristics, namely odor and color, as well as stability. The obtained cork powder extract exhibits antioxidant, anti-senescence, and anti-inflammatory properties.

[0015] The method of obtaining the developed extract is an ecological and sustainable method and easy to scale up to an industrial scale, since it allows it to be carried out at room temperature, allows the extraction solvent to be water and also allows the extract to be obtained with just one extraction cycle.

[0016] HPLC analysis of the cork extract concluded that it has a high phenolic content. The safety profile of the extract in this disclosure was also confirmed in cells representative of human epidermis and reconstructed human epidermis (Episkin). The source material also has a reproducible profile, representing consistent ingredient quality. Cosmetic / pharmaceutical. The extract in this disclosure demonstrated antioxidant, anti-inflammatory, and anti-aging effects, demonstrating application in the prevention and treatment of inflammation. Furthermore, the extract was found to have adequate microbiological quality and stability. The overall results demonstrate the unique properties of this extract and its application as an active ingredient for incorporation into a cosmetic product or for use as a medicinal product.

[0017] The extract of the present disclosure is unique, since this aqueous extract of cork powder showed antioxidant potential by increasing the expression of the antioxidant genes NQO1 (NAD(P)H Quinone Dehydrogenase 1) and HMOX1 (Heme Oxygenase 1), and increased the levels of HMOX protein (Heme Oxygenase) per se and under an inflammatory environment (macrophages exposed to LPS (lipopolysaccharide). This extract, in an inflammatory environment (i.e., cells exposed to LPS) induced a significant reduction in the expression of the Nos2 (Nitric Oxide Synthase 2) gene, reinforcing the anti-inflammatory potential of the present aqueous extract of cork powder.

[0018] One aspect of the present disclosure relates to a cork powder for use in cosmetics or medicine (human or veterinary) that comprises cork particles with a particle size of less than 100 pm, and in which the color presents L* values between 45-55; a* between 6-10 and b* between 12-16, measured by the CIELAB system.

[0019] In a preferred embodiment, the cork powder comprises cork particles with a particle size of less than 63 pm; preferably ranging from 16-63 pm; more preferably 20-40 pm.

[0020] In a preferred embodiment, the cork powder is a powder that has been subjected to 180°C for at least 30 minutes; preferably 30 minutes (sterilization process).

[0021] In a preferred embodiment, the oil absorption capacity of cork powder is at least 4 times greater than that of kaolin (measured in g / 100g by ISO 787 / 5-1980).

[0022] Another aspect of the present disclosure relates to a method for obtaining cork powder comprising the following steps: providing cork powder comprising cork particles with a particle size of less than 100 pm; preferably less than 63 pm; more preferably ranging from 16-63 pm; even more preferably ranging from 20-40 pm; adding water and stirring for at least 15 minutes (preferably 15-20 minutes at 1250 rpm) in order to obtain a cork powder dispersion; preferably wherein the mass ratio of cork powder (g): aqueous solvent (mL) ranges from 1:10 to 1:20; more preferably 1:16; filtering the cork powder dispersion through a porous glass filter in order to obtain clean cork powder; preferably the porous glass filter is of the G4 type; drying the clean cork powder at at least 50 °C for at least 1 h, in order to obtain dry cork powder; heating the dry cork powder at 180 °C for at least 30 minutes in order to obtain the cork powder as described in the present disclosure.

[0023] In a preferred embodiment, the cork powder of the method described herein comprises cork particles with a particle size ranging from 16-63 pm; preferably 20-40 pm.

[0024] The cork powder described in this disclosure can be obtained at various stages of cork processing. In a preferred embodiment, the cork powder of the present disclosure is selected from grinding dust; cleaning dust; powder from particle size separations; powder from particleboard panel finishing; powder from cork stopper and disc finishing; powder from natural cork stopper finishing; or a mixture thereof. Preferably, the powder of the present disclosure is obtained at the particle size separation stage (granul size separation powder).

[0025] Another aspect of the present disclosure relates to an aqueous extract of cork powder for cosmetic or medical use comprising: castalagin, in a concentration greater than 10 pg / mg (dry extract); gallic acid, in a concentration greater than 2 pg / mg (dry extract); ellagic acid, in a concentration greater than 4 pg / mg (dry extract).

[0026] In a preferred embodiment, the extract is obtained from the cork powder described in the present disclosure, preferably from a cork powder comprising cork particles with a particle size of less than 100 pm, and in which the color presents L* values between 45-55; a* between 6-10 and b* between 12-16, measured by the CIELAB system.

[0027] In a preferred embodiment, the aqueous extract of cork powder is an aqueous extract of lyophilized cork powder.

[0028] In a preferred embodiment, the aqueous extract of cork powder comprises: castalagin, in a concentration ranging from 10-60 pg / mg (dry extract), preferably from 10-50 pg / mg (dry extract); gallic acid, in a concentration ranging from 2-20 pg / mg (dry extract), preferably from 2-15 pg / mg (dry extract); ellagic acid, in a concentration ranging from 4-30 pg / mg (dry extract), preferably from 4-20 pg / mg (dry extract).

[0029] In one embodiment, the color of the extract has an L* between 40-60; an a* between 2 and 8 and a b* between 10-20, measured by the CIELAB system.

[0030] Another aspect of the present disclosure relates to a cosmetic or pharmaceutical composition comprising the cork powder as described in the present disclosure or the aqueous extract of cork powder as described in the present disclosure.

[0031] In a preferred embodiment, the composition comprises 0.1-10% (w / w) cork powder. In a preferred embodiment, the composition comprises 1-7% (w / w) cork powder; more preferably 4-6% (w / w) cork powder; even more preferably 5% (w / w).

[0032] In a preferred embodiment, the composition comprises 0.1-4% (w / w) cork powder; preferably 1-3% (w / w) cork powder; more preferably 2% (w / w).

[0033] In one embodiment, the composition further comprises titanium dioxide; preferably 1-15% (w / w) titanium dioxide; more preferably 8-12% (w / w).

[0034] Surprisingly, a synergistic effect was observed between the cork powder of this disclosure and titanium dioxide, in which the mixture of these two components allows for a higher sun protection factor (at least twice as high).

[0035] In one embodiment, the composition comprises 1-7% (w / w) cork powder; preferably 5% (w / w); and 1-15% (w / w) titanium dioxide; preferably 10% (w / w).

[0036] In a preferred embodiment, the composition further comprises: a mixture of glyceryl monostearate, cetostearyl alcohol, cetyl palmitate and coco-glycerides; preferably 2-8% (w / w); ethoxylated cetostearyl alcohol (ceteareth-12); preferably 0.1-3% (w / w); medium chain triglycerides; preferably 4-12% (w / w); cetearyl isononanoate; preferably 4-12% (w / w).

[0037] In a preferred embodiment, the sun protection factor of the composition is at least 40, preferably at least 50; more preferably at least 60, even more preferably between 40-80. (measured by method described in the scientific literature: Pissavini M, Tricaud C, Wiener G, Lauer A, Contier M, Kolbe L, Trullás Cabanas C, Boyer F, Nollent V, Meredith E, Dietrich E, Matts PJ. Validation of an in vitro sun protection factor (SPF) method in blinded ring-testing. International Journal of Cosmetic Science, 2018 Apr 20. doi: 10.1111 / ics.12459).)

[0038] In a preferred embodiment, the composition further comprises: 2-8% (w / w) of a mixture of polyglyceryl-6 esters of olive oil, sodium stearoyl lactate, cetostearyl alcohol; 4-12% (m / m) dicapryl ether; 1-4% (w / w) squalene; 1-3 (m / m) of caprylic / capric acid triglycerides; 1-3% (m / m) stearic acid; 1-3% (w / w) sweet almond oil; 0.1-0.5 (m / m) Xanthan gum 1-3%(w / w) Propanediol 2-8% (w / w) Glycerin 0.1-2% (w / w) of a mixture of phenoxyethanol, benzyl alcohol, ethylhexylglycerin and tocopherol.

[0039] In a preferred embodiment, the color of this cosmetic composition has an L* between 40-54; an a* between 4-8 and a b* between 10-17, measured by the CIELAB system.

[0040] The cork powder and extract of this disclosure are compatible with the skin, not exhibiting irritant potential (assessed by the OECD 439 test).

[0041] Another aspect of the present disclosure relates to a cosmetic or pharmaceutical composition comprising the aqueous cork powder extract of the present disclosure. In a preferred embodiment, the composition comprises 1-2% (w / w) aqueous cork powder extract. In an even more preferred embodiment, the composition comprises 0.1-2% (w / w) aqueous cork powder extract; preferably 0.1-1% (w / w); more preferably 0.5-1% (w / w).

[0042] In one embodiment, the composition comprising the aqueous extract of cork powder further comprises: 4-6% (w / w) of a mixture of polyglyceryl-6 esters of olive oil, sodium stearoyl lactate, cetostearyl alcohol; 10-20% (w / w) of triglycerides of capric acid and / or caprylic acid; 1-3% (w / w) cetostearyl alcohol; 0.1-0.2 (m / m) xanthan gum; 1-3% / (w / w) propylene glycol; 0.1-2% (w / w) of a mixture of phenoxyethanol, benzyl alcohol, ethylhexylglycerin and tocopherol.

[0043] In a preferred embodiment, the color of the composition comprising the aqueous extract of cork powder has an L* between 48-55; an a* between 2 and 7 and a b* between 15-20, measured by the CIELAB system.

[0044] The composition of the present disclosure may be presented in different pharmaceutical forms. In a preferred embodiment, the composition of the present disclosure is selected from a list comprising: solution, serum, cream, lotion, gel, hydrogel, oleogel, oil, soap, shampoo, dry shampoo, spray, ointment, paste, mousse, or body wash. In an even more preferred embodiment, the composition is a cream.

[0045] Another aspect of the present disclosure relates to the use of cork powder or the composition comprising cork powder as an oil-absorbing agent.

[0046] Another aspect of the present disclosure relates to the use of cork powder or the composition comprising cork powder as a pigment; preferably as a cosmetic pigment.

[0047] Another aspect of the present disclosure relates to the use of cork powder or the composition comprising cork powder as a sun protection factor enhancing agent, preferably wherein the enhancement factor is at least 2 times; more preferably the enhancement factor ranges between 2-3 times.

[0048] Another aspect of the present disclosure relates to the use of cork powder or the composition comprising cork powder as an ultraviolet radiation blocking agent.

[0049] Another aspect of the present disclosure relates to the use of cork powder or the composition comprising cork powder in medicine or veterinary medicine. In a preferred embodiment, the cork powder or composition is used in the prevention and treatment of skin diseases, preferably skin diseases caused by sun exposure. In another embodiment, the cork powder or composition is used in the prevention and / or treatment of skin cancer.

[0050] Another aspect of the present disclosure relates to the use of the extract or the composition comprising the extract as a preventive or retarding agent for skin aging.

[0051] Another aspect of the present disclosure relates to the use of the extract or the composition comprising the extract as an anti-senescent (anti-aging) agent.

[0052] Another aspect of the present disclosure relates to the use of the extract or the composition comprising the extract as an antioxidant agent.

[0053] Another aspect of the present disclosure relates to the use of the extract or the composition comprising the extract for medicinal use. In a preferred embodiment, it relates to the use of the extract or the composition comprising the extract either for use in the prevention or treatment of inflammation, and / or for use as an anti-inflammatory; preferably a topical anti-inflammatory.

[0054] Another aspect of the present disclosure relates to a method for obtaining the extract described herein comprising the following steps: providing cork powder comprising cork particles with a particle size of less than 0.5 mm; adding to the cork powder an aqueous solvent in a mass ratio of cork powder (g):aqueous solvent (mL) ranging from 1:20 to 1:80, and stirring for at least 30 minutes to obtain a mixture comprising the aqueous extract of cork powder; filtering the mixture comprising the aqueous extract of cork powder to obtain an aqueous extract of cork powder.

[0055] In a preferred embodiment, the method comprises an additional step of drying and lyophilizing the aqueous extract of cork powder in order to obtain a lyophilized aqueous extract of cork powder.

[0056] In a preferred embodiment, the granulometry of the cork particles to obtain the extract is less than 250 pm; preferably less than 200 pm; more preferably between 63-200 pm; most preferably 100-200 pm.

[0057] In a preferred embodiment, the granulometry of the cork particles to obtain the extract is less than 100 pm.

[0058] In a preferred embodiment, the mass ratio of cork powder: aqueous solvent ranges from 1:30 to 1:50. In an even more preferred embodiment, the mass ratio of cork powder: aqueous solvent is 1:40.

[0059] In a preferred embodiment, the agitation rate of the aqueous solvent and cork powder ranges from 500-1000 rpm; preferably 700 rpm.

[0060] In a preferred embodiment, the aqueous solvent is water

[0061] In a preferred embodiment, the method is carried out at room temperature (20-25°C).

[0062] In a preferred embodiment, the stirring time of the aqueous solvent and cork powder ranges from 30 - 120 minutes; preferably 60 minutes.

[0063] In a preferred embodiment, the filtration step is carried out with a glass fiber filter membrane; preferably with a diameter of 47 mm and a pore size of 1.2 pm.

[0064] In a preferred embodiment, the extraction step (adding to the cork powder an aqueous solvent in a mass ratio of cork powder:aqueous solvent ranging from 1:20 to 1:80, and stirring for at least 30 minutes, to obtain a mixture comprising the aqueous extract of cork powder) is carried out in a single extraction cycle.

[0065] The cork powder used to obtain the cork powder extract may originate from different stages of cork processing. In a preferred embodiment, the cork powder is selected from: grinding powder, cleaning powder, powder from particle size separations, powder from particleboard panel finishing, powder from cork stopper and cork disc finishing, powder from natural cork stopper finishing, or a mixture thereof. In an even more preferred embodiment, the cork powder is powder from particle size separations. BRIEF DESCRIPTION OF THE FIGURES

[0066] For easier understanding, the figures are attached, which represent preferred embodiments that are not intended to limit the scope of this description.

[0067] Figure 1: Production Process and Origin of the By-products under study.

[0068] Figure 2. Powder from particle size separations, preferably obtained in the process of dedusting and transporting granulate, with particle size between 0-2 mm. Also called PT powder (transport powder) or Pl powder).

[0069] Figure 3. NIR (a) and MIR (b) spectra of P0 powder and Pl powder.

[0070] Figure 4. Score plots obtained from PCA (Principal Component Analysis) using NIR spectra of different batches of P0 and Pl powders preprocessed with the Savitzky-Golay filter (15-point filter width, second-degree polynomial, first derivative) followed by standard normal variant (SNV) and mean center.

[0071] Figure 5. Incorporation of different by-products into a semi-solid formulation.

[0072] Figure 6. Determination of the FPS enhancing capacity of CIE powder.

[0073] Figure 7. Analysis of the chemical profile of Cl powder and sterilized Cl powder (CIE), a) NIR; b) MIR.

[0074] Figure 8. Mixture of different concentrations of Cl with pigments used in decorative cosmetics, a) 1% (m / m) CIE powder + 2% (m / m) red iron oxide + 4.5% (m / m) Titanium Dioxide; b) 2% (m / m) CIE powder + 5.5% (m / m) Titanium Dioxide; c) 5% (m / m) CIE powder + 2.5% (m / m) Titanium Dioxide.

[0075] Figure 9. Examples of several different formulations tested, containing different concentrations of cork powder.

[0076] Figure 10. Viscosity curve of the base formulation of a colored cream.

[0077] Figure 11. Texturogram of the spreadability test of the basic formulation with CIE powder stored at 25°C (black) and 40°C (blue) for 15 days (n=l).

[0078] Figure 12. Skin irritation test results.

[0079] Figure 13. Appearance of Pl powder.

[0080] Figure 14. CIE powder color analysis over 90 days at two different temperatures: 25 and 40°C.

[0081] Figure 15. Evolution of CIE powder pH over 90 days of storage at two different temperatures: 25 and 40°C. n=3.

[0082] Figure 16. Analysis of the chemical profile of CIE powder over 90 days and stored at different temperatures: 25 and 40°C - through Near Infrared Spectroscopy (NIR).

[0083] Figure 17. Analysis of the chemical profile of CIE powder over 90 days and stored at different temperatures: 25 and 40°C - through Mid-Infrared Spectroscopy (MIR).

[0084] Figure 18. Analysis of the moisture content of CIE powder over 90 days and stored at different temperatures: 25 and 40°C (n=3).

[0085] Figure 19. Analysis of the stability of the formulation with CIE powder, for 90 days, at 25 and 40°C - Appearance and odor.

[0086] Figure 20. Color parameters after storage of the BBcO5 formulation for 90 days at 25 and 40°C - Color (n=3).

[0087] Figure 21. Analysis of the pH of the formulation with CIE powder, for 90 days, at 25 and 40°C (n=3).

[0088] Figure 22. Analysis of the stability of the BBcO5 formulation for 90 days, at 25 and 40°C with regard to flow.

[0089] Figure 23. Variation of extract mass with the number of extractions.

[0090] Figure 24. Masses obtained for the different extracts (A); Evaluation of the extraction effectiveness according to the absorbance of the extract at 280 nm (B) and 350 nm (C).

[0091] Figure 25. DPPH radical scavenging activity. Results expressed as mean ± standard error, n = 3; ascorbic acid (positive control): IC5o = 7.9 ± 0.4 pg / mL.

[0092] Figure 26. Effect of cork extracts on cellular metabolism. Macrophages (A), fibroblasts (B), and keratinocytes (C) were cultured and exposed to different concentrations of cork extracts for 24 h. The Alamar blue assay was also performed to evaluate cellular metabolism. Data correspond to the mean ± SEM of at least three independent experiments and are represented as % of control cells (Ctr, black bars).

[0093] Figure 27. Effect of cork extracts on NO production in macrophages stimulated with the pro-inflammatory mediator LPS. Cells were plated and exposed to 62.5 and 125 pg / mL of cork extracts for 24 h, in the presence or absence of LPS (100 ng / mL). The Griess assay (graphs letter A) and the Alamar blue assay (graphs letter B) were performed to evaluate nitrite levels in the supernatant and in cellular metabolism (after LPS exposure), respectively. Data correspond to the mean ± SEM of at least three independent experiments and are represented in nitrite concentration (pM).

[0094] Figure 28. Effect of cork extracts on gene expression related to the inflammatory response. Macrophages were cultured and exposed to 125 pg / mL of cork extracts, and to commercial compound Oleoactif, in the presence or absence of LPS (100 ng / mL) for 6 h. The expression of the pro-inflammatory genes Nos2, I lip, Tnfa, 116 and the anti-inflammatory gene 1110 was determined by real-time RT-PCR. The data correspond to the mean ± SEM of at least three independent experiments and are expressed relative to control cells (Log2=0).

[0095] Figure 29. Effect of cork extracts on the expression of genes encoding antioxidant proteins. Keratinocytes were cultured and exposed to 125 pg / mL of cork extracts and the commercial compound Oleoactif for 6 h. The expression of the antioxidant genes HMOX1, NQ01, and TRX was determined by real-time RT-PCR. Data correspond to the mean ± SEM of at least three independent experiments and are expressed relative to control cells (Log2=0).

[0096] Figure 30. Effect of cork extracts on the levels of the pro-inflammatory enzyme iNOS. Macrophages were cultured and exposed to 125 pg / mL of Pl cork extracts and the commercial compound Oleoactif, in the presence or absence of LPS (100 ng / mL) for 24 h. iNOS protein levels were determined by Western blotting. Representative images of the blots are shown. Data correspond to the mean ± SEM of at least three independent experiments and are expressed as % LPS.

[0097] Figure 31. Antioxidant effect of cork extracts on immune cells. Macrophages were cultured and exposed to 125 pg / mL of cork extracts and the commercial compound Oleoactif, in the presence or absence of LPS (100 ng / mL) for 24 h (A) or rotenone (20 pM) for 6 h (B). (A) Levels of the antioxidant enzyme HMOX1 were determined by Western blotting. Representative images of the blots are shown. (B) Superoxide anion (O2 ) levels were determined by the number of Mitosox red-positive cells, assessed by flow cytometry. Data correspond to the mean ± SEM of at least three independent experiments and are expressed as % of LPS (A) or Rot (B).

[0098] Figure 32. Effect of cork extracts on skin regeneration. Fibroblasts were cultured and exposed to 125 pg / mL of cork extracts and the commercial compound Oleoactif for 6 h (A) or 18 h (B). (A) The expression of the genes encoding the proteins collagen type I (Collal) and Integrin beta-1 (Itgbl) was determined by real-time RT-PCR. (B) The scratch wound assay was performed to evaluate the effect of cork extracts on cell migratory capacity and skin regeneration. The area occupied was calculated and the results represented in the graph. Representative microscopic images are illustrated in the panel (scale bar = 50 μm). The data correspond to the mean ± SEM of at least three independent experiments and are expressed relative to the control (Ctr).

[0099] Figure 33. Anti-senescence effect of cork extracts. Fibroblasts were cultured and exposed to etoposide (Ctr+; 12.5 pM), a senescence inducer, for 24 h, and the cells were subsequently subjected to a recovery period in the presence of 125 pg / mL of Pl cork extracts and the commercial compound Oleoactif for 72 h. Cellular senescence was assessed using a kit commercially available beta-galactosidase staining. The number of senescent cells (stained blue due to beta-galactosidase activity) was visualized by microscopy. Representative microscopic images are shown (scale bar = 20 μm). Data correspond to the mean ± SEM of at least three independent experiments and are expressed relative to the control (Ctr).

[0100] Figure 34. Effect of aqueous PI extract on the EpiSkin skin irritation test.

[0101] Figure 35. NIR spectrum of the aqueous PI extract.

[0102] Figure 36. MIR spectrum of aqueous PI extract.

[0103] Figure. 37. Record of the appearance of the aqueous PI extract over time (time (T) in days).

[0104] Figure 38. Variation of the chromatic coordinates of the aqueous PI extract over time at temperatures of 25 °C and 40 °C (time (T) in days).

[0105] Figure 39. Variation of pH of the aqueous PI extract over time at temperatures of 25 °C and 40 °C (time (T) in days).

[0106] Figure 40. Overlay of NIR and MIR spectra of the aqueous PI extract obtained throughout the stability test at 25 °C and 40 °C (time (T) in days).

[0107] Figure 41. Evolution of the DPPH uptake activity (IC5o) of the aqueous extract over the storage time (n=3) (time (T) in days).

[0108] Figure 42. Evolution of the DPPH uptake activity (IC5o) of the hydroglyceric solution over the storage time at 25 °C, n=3 (time (T) in days).

[0109] Figure 43. Evolution of the pH of the aqueous extract in hydroglyceric solution (time (T) in days).

[0110] Figure 44. Evolution of DPPH uptake activity (IC5o) and pH of the aqueous extract in pH 5 and 7 buffer solutions stored at 25 °C. n=3 (time (T) in days).

[0111] Figure 45. Texturograms of CR002 cream stored for 15 days at 25 (A) and 40 °C (B).

[0112] Figure 46. Luminosity and chromatic coordinates of the CR002 formula throughout the stability test (time (T) in days).

[0113] Figure 47. pH variation of the CR002 formulation over time (days).

[0114] Figure 48. Apparent viscosity of formulation CR002 with aqueous extract. DETAILED DESCRIPTION

[0115] This disclosure relates to a cork powder and an aqueous extract thereof, and a method for obtaining said powder and said extract. Furthermore, this application relates to the use of the extract in cosmetics or medicine.

[0116] In the prior art, color can be measured by several methods. In the present disclosure, color was measured by the CIE / .AB (or CIE L*a*b*) system. L* is the lightness value (which defines black as 0 and white as 100), the chromatic coordinate a* corresponds to the colors green-red (where negative values correspond to green and positive values to red), and the chromatic coordinate b* corresponds to the colors blue-yellow (where negative values correspond to blue and positive values to yellow).

[0117] Cork powder particle size can be measured in several ways. In this publication, particle size was measured using the laser diffraction method using the Mastersizer 3000 equipment, or through the standardized particle size analysis method using mechanical sieving. Specifically, regarding the particle sizes obtained, sieves with mesh openings of 1000 pm, 800 pm, 180 pm, 63 pm, and / or 16 pm were used. Valorization of cork powder for cosmetic applications

[0118] A study was carried out to identify from which of the cork processing phases the cork powder should be collected, as well as the methodologies required for its treatment, with a view to its valorization as a cosmetic ingredient (Figure 1). Collection of cork dust at different extraction points

[0119] The cork processing phases were identified to determine which cork powder should be collected for the new cosmetic product. Dust was collected during the crushing, drying of the crushed and granulated cork, dust removal, and transportation of the granulated cork, and finally, during the granulated sterilization phase. Treatment of cork powder with a view to its valorization

[0120] The cork processing was tailored to the characteristics required for the extracted powder. Thus, the following processes were carried out:

[0121] -Crushing process: crushing cork planks and waste into pieces between 2 and 35 mm. This process produced powder between 0 and 2 mm in size, mostly consisting of cork back. PG cork powder, hereinafter referred to as P0, was obtained at this stage of the process. Heterogeneity was observed in both grain color and size. Density and moisture content were analyzed, with values of 287 kg / m 3 and 40%, respectively. TCA was also determined, with a value of 13.7 ng / L.

[0122] Crushed cork drying process: The crushed cork drying process consists of passing the crushed cork through two rotating drums continuously for 10 minutes. Drying is achieved by radiation generated by water vapor. During drying, the powder grains are released from the crushed cork and are sucked into the dryer's dust extraction filter. PS cork powder was obtained at this stage of the process.

[0123] - Crushed cork grinding process - GRANULATION: The granulation process consists of crushing the crushed cork into grains with dimensions between 0.2 and 8 mm. This process created powder between 0 and 0.2 mm. The PT cork powder, hereinafter referred to as Pl, was obtained at this stage of the process (Figure 2), more specifically in the final stage of the process, in the separation of the granules by different particle sizes (powder from particle size separations). Cosmetic formulations comprising cork powder

[0124] Cosmetic makeup and photoprotective product formulations comprising cork powder were developed to demonstrate the cosmetic applicability of this natural ingredient. Pre-formulation and formulation of cosmetic makeup products with photoprotective properties based on cork powder

[0125] Characterization studies of cork powder were carried out, which included the following tests: moisture content, oil absorption, color and particle diameter.

[0126] To assess moisture content, an infrared balance was used, where the samples were subjected to a temperature of 80°C for 10 minutes. To determine apparent density, a 250 ml graduated cylinder was used, into which an appropriate mass of powder sample was placed. Tests were performed in triplicate for each sample, without beating / compacting.

[0127] To determine the color of the powder samples, the Chromameter CR-400 colorimeter was used, where tests were carried out for each of the powder samples, in triplicate, under the following conditions: - Observer degree: 2 degrees - Primary light source: D65

[0128] Samples were analyzed in small plastic Petri dishes

[0129] Particle diameter was determined using Malvern's Mastersizer 3000 equipment. Samples were previously prepared for analysis on the equipment.

[0130] Additionally, the profile of volatile compounds that give the powders their characteristic odor was characterized by HS-SPME (Table 1). This test allowed the detection and Identification of 10 compounds within the classes of ketones, aromatic aldehydes, monoterpenes, and alcohols allowed us to assess possible differences and similarities between the profiles of the PO and Pl powders. Semi-quantitative analysis revealed a similar volatile compound profile in the PO and Pl powders. The PO powder contained phenylethyl alcohol, which was not present in the Pl powder, as well as slightly elevated concentrations of pent-2-one, 4-methylpent-2-one, hex-2-one, and vanillin. Limonene was only detected in the Pl powder. Camphene and borneol were present in the Pl powder in slightly higher concentrations, and eucalyptol and camphor in much higher concentrations than in the PO powder.

[0131] Table 1 - Analysis of volatile compounds of PO and Pl powders.

[0132] The analysis of the volatile compound profile of PO and Pl cork powders revealed their potential as a sustainable source for fragrance production and, consequently, for contributing to a pleasant aroma in formulations to which they are added. Surprisingly, PI powder proved to be a preferable source of volatile compounds, presenting a higher monoterpene content than PO powder. Through HS-SPME analysis, it was possible to determine the volatile compounds that contribute to the organoleptic properties of the powders. While PO powder presented slightly higher levels of vanillin, PI powder presented much higher amounts of monoterpenes in the HS-SPME analysis.

[0133] To analyze changes in the chemical profile of three batches of P0 and Pl powders, NIR spectra were obtained (Figure 3). The results revealed that the P0 batches are quite similar to each other, as was also observed in the Pl batches. The spectra of the P0 powder batches presented more intense bands in the range of 7000 to 5200 cm 1 (water band region), which allowed us to conclude that the P0 powder has a greater amount of water than the Pl powder. However, the analysis of the raw NIR spectra of the The fact that the sample size was insufficient to ensure chemical similarity between the samples, since many different chemical bonds could absorb in the same wavenumber region. Therefore, a PCA was performed to verify the difference between the PO and Pl powders. Analysis of the PCA results (Figure 4) using the NIR spectral data revealed that the cork powder samples tend to form clusters according to sample type. Samples from the PO batches obtained negative scores in principal component 1 (PC 1), while samples from the Pl batches obtained positive scores. This fact reinforces the idea that the NIR spectra of the PO and Pl powders are different, while the samples from each batch are similar.

[0134] "PL powder" was selected for further studies, as it presented characteristics most favorable for use as a cosmetic pigment. Therefore, a reproducibility study was conducted on this powder to determine whether the same byproduct produced on different days had the same characteristics. Tests were performed on moisture content, color, and particle diameter. For a more extensive characterization of the PL powder, the following tests were performed: pH, compressibility, oil absorption capacity, heavy metal content, and microbiological quality.

[0135] Tests on samples of Pl powder from different days were carried out in order to verify the reproducibility of the batches. The variability of the different batches was less than 15%, confirming adequate reproducibility (Table 2). Table 2 - Batch reproducibility assessment tests for Pl powder

[0136] For the selected Pl powder, additional tests were carried out to enhance the value of this by-product, namely the determination of pH, flow and compaction properties, oil absorption capacity, heavy metal content and microbiological quality.

[0137] To determine the pH, a 10% (w / w) aqueous dispersion with Pl powder was prepared under magnetic stirring for 10 minutes. The suspension obtained was filtered with a filter paper, and the pH was measured in the filtrate with a Hl 2211 pH / ORP / °C meter (Hanna Instruments, Woonsocket, RI, USA). Measurements were performed in triplicate (Table 3).

[0138] Table 3 - Determination of pH of Pl powder.

[0139] To determine the flow properties, namely the compressibility and strength of the compact obtained with Pl powder, the European Pharmacopoeia 10 test and a TA.XT.plus texturometer from Stable Micro Systems were used, respectively (Table 4).

[0140] Table 4 - Determination of the compressibility index of powder Pl

[0141] The oil absorption capacity of Pl powder was studied in accordance with ISO 787 / 5-1980. lg of Pl powder was placed on a glass plate. Crude linseed oil was added dropwise to the Pl powder. After each addition, the oil was thoroughly incorporated into the Pl powder with a metal spatula until it spread without cracking or crumbling. The same process was performed on lg of kaolin, frequently used in cosmetic formulations for its oil absorption capacity. The weight of linseed oil that must be absorbed by a given weight of cork powder is defined as the comparative oil absorption value (Table 5).

[0142] Table 5 - Determination of the oil absorption value of the Pl powder, compared with the control.

[0143] To determine heavy metal content, Pl powder samples were solubilized by microwave-assisted closed-vessel acid digestion in an ETHOS EASY microwave oven equipped with an SK-15 easyTEMP high-pressure rotor, following a procedure based on the U.S. Environmental Protection Agency (EPA) method 3052 (Microwave-assisted acid digestion of siliceous and organic-based matrices). Blank samples were obtained using the same procedure. For analytical quality control purposes and considering the type of sample analyzed in this study, two certified reference materials (CRMs) were used: Cabbage Powder (BCR-679) and Bladderwrack (Fucus vesiculosus) Powder (ERM-CD200), both from the European Commission's Joint Research Centre, subjected to the same sample pretreatment. Samples were analyzed in triplicate, and a blank assay was performed in each digestion batch.Trace element determination was performed by inductively coupled plasma mass spectrometry (ICP-MS) (Table 6). Table 6 - Determination of heavy metal content (mean ± SD) in Pl(ppm) n=3

[0144] To optimize the characteristics of the Pl powder, which proved to be the most promising, it was processed. To this end, the Pl powder was sieved through sieves with 180 pm and 63 pm mesh openings. The fraction above 63 pm (fraction E) was collected to obtain bioactive extracts, and the fraction below 63 pm (fraction C) was used to obtain makeup pigments and conduct preformulation studies on this type of product.

[0145] Subsequently, fraction C was also processed to obtain a more homogeneous powder with a smaller particle diameter. First, a portion of fraction C (25 g) was suspended in 400 mL of water with magnetic stirring (1250 rpm) for 20 minutes. The suspension was then filtered through a G4 crucible filter with a 16 μm pore size. The fraction retained on the filter surface was collected and dried in a fluid-bed oven at 50 °C for 1 hour, yielding fraction C1, with a particle diameter between 16 and 63 μm. To avoid waste, the filtrate obtained is also used for incorporation into semi-solid formulations.

[0146] The Cl fraction is the powder fraction that was used for preformulation studies as a pigment in makeup products. Extensive characterization of this fraction was performed through tests such as moisture content, particle diameter, apparent density, pH, color, oil absorption, hygroscopy, water-soluble content, and volatile matter at 105°C (Table 9).

[0147] Hygroscopy was assessed according to the European Pharmacopoeia 10. The weighing vessel and stopper were weighed (mj. Briefly, 1.5 g of Cl powder was placed at the bottom of the weighing vessel, which was subsequently stopped and weighed (m2). The non-perforated weighing vessel was placed in a desiccator containing a saturated ammonium sulfate solution (relative humidity = 78.6%) at 25°C for 24 h. The weighing vessel was stopped and weighed (m3).

[0148] The water-soluble content of Cl powder was determined according to ISO 787-3:2000. 5 g of Cl powder (mo) was suspended in 200 mL of boiling water and stirred for 5 min. The mixture was then transferred to a 250 mL volumetric flask and diluted with water to the mark. The filtrate was placed in a porcelain dish, and the residue was dried in an oven at 105°C for 2 h. The process was repeated until two consecutive weighings differed by no more than 10% (mj).

[0149] The volatile matter of Cl powder at 105°C was studied in accordance with ISO 787-2:2021. 10 g of Cl powder was placed in a uniform layer at the bottom of the weighing bottle. The weighing bottle was placed in an oven at 105°C for one hour. The process was repeated up to two

[0150] Next, the microbiological quality of the PI powder and the Cl fraction was assessed. In this context, it is worth noting that the microbiological quality analysis protocol was prepared in accordance with ISO 21148:2017 and the European Pharmacopoeia 10, specifically the chapter "Microbiological examination of non-sterile products." The sample was prepared by diluting lg of PI powder in l0mL of saline. Serial dilutions were also prepared up to 10 -3. Aliquots of 100pL of the original samples and dilutions were spread, in duplicate, on the surface of Tryptic Soy Agar (TSA) medium for bacterial isolation, and incubated for 24-48 hours at 30-35 ± 2°C. Subsequently, bacterial isolates were identified based on the Gram staining method. The detection of specific microorganisms, which should be absent in the studied sample - E. coli, S. aureus, P. aeruginosa and C. albicans, followed in the respective selective media, according to a protocol defined based on ISO standards and the European Pharmacopoeia 10. In order to identify the Enterobacteriaceae present in the Pl powder sample, according to their biochemical characteristics, an API (Analytical Profile Index) 20 E was performed. An ID 32 C was also performed to identify the yeasts present in the Pl powder sample, according to their biochemical characteristics.

[0151] Table 7 - Determination of the microbiological quality of the Pl powders and Cl fraction through the counting of total aerobic microorganisms and the identification of specific microorganisms

[0152] However, as the total count of aerobic microorganisms exceeded the maximum limit defined in ISO 17516:2014 and in the European Pharmacopoeia 10 (Table 7), thermal or chemical treatments of the Cl powder were carried out under different conditions, in order to understand which were the least aggressive conditions that would allow the optimization of the microbiological quality of the ingredient to a value within the regulatory limits (less than 1.00xl0 3 CFU / g). Various methods were implemented with the aim of reducing the microbiological load of the powder Cl -Table 8. Table 8 - Optimization of the microbiological quality of Cl powder

[0153] It was found that changing the processing of the Cl powder, suspending it in a 15% hydroalcoholic mixture instead of water, was not effective in reducing the microbiological load. On the other hand, sterilizing the Cl powder at 180°C for 2 h, 1 h, and 30 min completely eliminated the microbiological load. Despite a considerable reduction in the microbiological load, the sterilization process at 180°C for 15 min did not ensure the required microbiological quality. Therefore, less aggressive conditions that improve microbiological quality to a lower value al.00xl0 3 CFU / g correspond to heating Cl powder to 180°C for 30 minutes. This sterilized Cl powder was given the CIE designation. CIE Powder Characterization

[0154] Next, the sterilized Cl powder (CIE) was characterized (obtained by heating the Cl powder to 180°C for 30 minutes). This is because, since the process for obtaining the Cl powder was modified by adding a heating step to 180°C (CIE), it was necessary to perform a new series of physical-chemical characterization tests on the powder to assess possible significant changes (Table 9). The tests on the Cl powder (non-sterilized) were repeated when analyzing the CIE powder, so that the values obtained in each test could be directly compared.

[0155] Table 9 - Physicochemical characterization tests of CIE powder, compared with Cl powder (non-sterilized)

[0156] Thus, no significant differences were observed between the parameters analyzed in the Cl and CIE powders. The moisture content and hygroscopy results presented the most disparate values. The CIE powder had half the moisture content and a hygroscopy value approximately six times higher than the Cl powder. This is due to the fact that the CIE powder was subjected to an extreme heating process, forcing the evaporation of the water incorporated into the sample.

[0157] Furthermore, in order to analyze any changes in the chemical profile of the Cl powder that may have been induced by the thermal process to which it was subjected, the Cl powder from a previous batch and the CIE powder were analyzed using spectroscopic methods—near-infrared (NIR) and mid-infrared (MIR) spectroscopy (Figure 7). NIR allows the exploration of the physical and chemical characteristics of solid samples, exploring the interactions between matter and light non-destructively and requiring only a small amount of CIE. The spectra recorded in the NIR region (10,000–4,000 cm 1 ) are sensitive to a wide variety of compounds and molecular interactions. In MIR, most functional groups can be observed, since their vibrational resonances lie within this range of excitation frequencies, and even complicated and similar structures, such as polysaccharides, can be differentiated. MIR complements NIR and covers the region 4000–400 cm 1and contains information about fundamental molecular vibrations. In hydrated samples, NIR spectra show two primary water absorption bands at frequencies of ~5200 and 7000 cm 1 . Comparing the graphs of the Cl and CIE powders, both NIR and MIR, it is possible to notice that there are differences due to the greater presence of water in the Cl powder than in the CIE powder (Figure 7). In other words, what is seen is not a change in the compounds present (since both graphs present the same peaks), but rather in the intensity of their bands.

[0158] Another study involved dispersing liquids used in cosmetics, attempting to facilitate powder dispersion in semi-solid or fluid formulations. Thirteen liquids commonly used for pigment dispersion in cosmetics were selected (propylene glycol, glycerin, castor oil, sweet almond oil, liquid paraffin, Crodamol IPIS, Ceraphyl 791, 1,3-Butylene Glycol, Hallbrite BHB, Evertouch™, Dimethicone Copolyol, 96% Alcohol, Cyclomethicone Tetramer), and then mixed with the powder. The mixtures were left to rest, and after 24 hours, their appearance was evaluated to determine which mixture was most homogeneous. The liquids chosen were propylene glycol and glycerin.

[0159] For pre-formulation studies, the by-products were first incorporated into a semi-solid formulation, namely an O / W emulsion. The P0, Pl, and Cl powders were tested, as well as the filtrate resulting from the processing of the Pl powder. The appearance of the formulations is shown in Figure 5.

[0160] Additionally, the ability to enhance the Sun Protection Factor (SPF) of CIE powder in semi-solid formulations called sunscreen emulsion (Table 10) was evaluated using the Labsphere UV-2000S Sunscreen Analyzer equipment.

[0161] Table 10: Anti-sun formulation.

[0162] The CIE powder was incorporated at 5% (w / w) into a formulation containing 10% (w / w) titanium dioxide, for which the SPF was determined. In parallel, the SPF of the same formulation containing only 10% (w / w) titanium dioxide was determined, serving as the control. The formulation containing CIE had an SPF value approximately 2.5 times higher than the control, confirming the SPF-promoting capacity of the Cl powder (Figure 6): - Sunscreen with 10% (w / w) titanium dioxide: 25.30 ± 2.54 (mean ± SD) - Sunscreen with 10% (m / m) titanium dioxide with 5% (m / m) powder Cl: 63.67±13.82 (mean ±SD) (2.5X higher than the control) Pre-formulation study of CIE Powder

[0163] Following the preformulation study previously initiated for all cork industry byproducts (P0 powder, PI powder, and Cl powder), preformulation studies of CIE powder followed, consisting of several blending tests of CIE powder with other pigments widely used in decorative cosmetic formulations, namely titanium dioxide and red iron oxide (Figure 8), at different concentrations. The objective was to obtain a relatively homogeneous, beige / brown mixture to produce a decorative makeup product (tinted cream). The ratio of CIE powder to titanium dioxide (Figure 8 c) is as close as possible to what was intended.

[0164] Next, different mixtures of CIE powder and titanium dioxide were incorporated into different base formulations (Figure 9). Formulation 5 (Table 11) was the one that presented the closest appearance to the desired one, so it was selected to proceed to experimental formulation studies.

[0165] Table 11 - Composition of formulation 5 (BBcO5) and respective preparation procedure

[0166] Formulation studies followed with the CIE powder (colored cream), involving the tests described below, relating to the basic formulation: Mechanical Stress Test, Rheological Characterization and Spreadability. Mechanical Stress Test

[0167] The centrifugation test subjects emulsions and suspensions to extreme gravitational forces to assess formulation stability, accelerating potential phase separation and sedimentation effects. Therefore, before starting a longer stability study, this test allows us to determine whether the base formulation is physically stable. If phase separation occurs, the cream is not stable, and therefore, the formulation will need to be modified before proceeding with the remaining tests.

[0168] In general, stress test conditions—rotations per minute (RPMs)—and centrifugation time vary depending on the study. However, the most common RPMs for cream centrifugation stress tests are 3,000–5,000 RPMs, and the most common centrifugation time is 30 minutes. Typical centrifugation settings are 3,500 RPMs for 30 minutes. Therefore, the selected base formulation was subjected to 3,500 RPMs for 30 minutes at 25°C. No distinct phases emerged; the formulation did not exhibit phase separation under the test conditions, demonstrating its stability. Rheological characterization of the base formulation

[0169] In this context, base formulations were selected to study the stability of the developed ingredients when incorporated into cosmetic product formulations. Considering the ingredient's intended purpose, a makeup base formula was selected (in which the Cl powder fraction was intended to be incorporated). The rheological behavior of the makeup base formulation was studied, particularly its flow. The formulation exhibited non-Newtonian fluid behavior of the pseudoplastic (rheo-fluidizing) type (Figure 10), i.e., the apparent viscosity decreased with increasing shear rate.

[0170] The behavior exhibited by the base formulation over time is confirmed when applying the power law model. The negative value of "n" indicates that the sample is more viscous at low shear rates, with the apparent viscosity decreasing as the shear rate increases (Table 12).

[0171] Table 12: Application of the power law model.

[0172] With the aim of studying the physical-chemical stability and activity of the ingredients over time, the respective stability evaluation protocols were also developed during this period, both for the ingredient (CIE fraction) and for the formulation. Spreadability

[0173] To assess the spreadability of the cream, a texturometer was used, to which a cone-shaped probe was attached and, as a base, another cone-shaped probe, into which the sample to be analyzed was inserted. The probe moves downward at a defined speed and distance.

[0174] The ease with which the conical probe moves through the cream translates into the product's spreadability. As soon as the probe comes into contact with the test sample, the measured force increases steadily until reaching a maximum value, the probe then reverses direction and withdraws from the sample. In this case, the measured force is negative because the formulation adheres to the probe, resisting its movement out of the sample container. It can be seen that storage temperature does not contribute to marked differences in the spreadability of the CIE powder formulation analyzed (Figure 11).

[0175] The CIE powder, due to its properties (low moisture content; adequate particle diameter; color; microbiological quality), was selected for pre-formulation studies (powder mixtures with pigments in different concentrations, dispersion in different solvents, among others) and formulation studies (stress test, rheological characterization, among others). parameters) using CIE powder as an ingredient, selecting a suitable base formulation for its incorporation, as well as the quantity of powder to be used. In vitro evaluation of cork powder toxicity

[0176] The toxicity of cork-derived powders, namely the PO, Pl and Cl fraction samples, was evaluated on a representative skin cell line (human keratinocytes). Cell viability was determined after exposing the cells to different concentrations (100, 50, 25, 10 and 5 mg / mL) of each sample.

[0177] It was possible to verify that cell viability approached 90% when the cells were exposed to cork powder samples at the following maximum concentrations: P0 = 50 mg / mL (95% viability); Pl = 10 mg / ml (94% viability) and Cl = 25 mg / ml (93% viability).

[0178] Finally, the irritant potential of the selected cork powder was assessed using a 3D model, Episkin, according to OECD Test Guideline 439. Briefly, the tissues were exposed to the CIE powder, the base formulation (formulation without Cl powder), and the base formulation with 5% Cl powder (Figure 12). Tissue viability was determined by the MTT assay. According to this skin irritation test, if the mean tissue viability is 50% or less, the analyte tested is considered irritating, which was not the case. Therefore, the Cl powder can be considered safe for use, as can the tested formulation incorporating Cl powder at a concentration of 5% (w / w) (Figure 12). Physical stability studies and characterization of quality parameters CIE Powder Stability Study

[0179] In order to study the physical-chemical stability and activity of the ingredient over time, a stability evaluation protocol was developed for the CIE powder (Table 13). Table 13 - Summary of CIE Powder Stability Protocol X: Test performed

[0180] The powder stability study was conducted over 90 days and was stored at two different temperatures: 25°C and 40°C. The storage container consisted of a PVP bag with a zipper closure, which came into direct contact with the powder, and which was placed inside a plastic container with two lids, minimizing the powder's contact with the external environment. Various physical, chemical, and microbiological parameters were analyzed throughout the analysis period.

[0181] Through the photographic records taken throughout the stability study, it can be concluded that, macroscopically, there were no changes in color or homogeneity. The woody odor of the CIE powder remained intense and constant throughout the analysis period.

[0182] At the same time, the color of the CIE powder was analyzed more objectively using the colorimeter, which allows obtaining information regarding the luminosity (L*) parameters and chromatic coordinates a* and b*. It was therefore confirmed that there are no considerable differences over the analysis time, regardless of the temperature (Figure 14).

[0183] The pH remained approximately constant throughout the analysis period (Figure 15). There was a slight drop in pH from time 30 onwards, but the parameter value remained within the range compatible with skin pH.

[0184] Regarding the spectrometric tests, there were some changes in the NIR in relation to the peaks that correspond to the zones ~5200 and 7000 cm 1 , which is possibly due to the fact that the Cl powder is hygroscopic and has the capacity to absorb water during the tests. The graphs of both NIR and MIR, for each temperature, remain practically constant over time, as understood by the almost total overlap at the different times (Figures 16 and 17).

[0185] The moisture content of the CIE powder was also evaluated throughout the 90-day test period. There was a slight increase in moisture content at time 90, which may be due to the powder absorbing some moisture during weighing for the test. Nevertheless, this parameter remained virtually constant throughout the analysis period (Figure 18).

[0186] Microbiological quality assessment was performed at the beginning of the test and 90 days after the start of the storage test at temperatures of 25°C and 40°C. At time 0, the powder was analyzed immediately after heating at 180°C for 30 minutes. The test was performed in a horizontal laminar flow cabinet to reduce possible risk of sample contamination. The CIE powder sample analyzed (consisting of a 1:10 dilution) was prepared and properly homogenized. Serial dilutions (10 1 , IO -2 and IO -3 ), in duplicate. The total count of total aerobic microorganisms was 0 CFU / g, 48h after the start of the analysis. The procedure was repeated 90 days after the stability study, both for the sample stored at 25°C, as for 40°C. In these cases, the count of aerobic mesophilic microorganisms was higher than that determined at time 0, but still lower than the limits established in ISO 17516:2014. Particle size and hygroscopy remained practically constant at times 0 and 90 (Table 14).

[0187] Table 14. - Results at=0 days, t=90 days (25 9 C) et=90 days (40 °C) Stability studies of the formulation with CIE powder

[0188] In order to study the physicochemical stability and activity of formulation 5 (BBcO5) over time, a stability evaluation protocol was developed for the CIE powder (Table 15). Table 15: Summary of the stability protocol of the formulation with CIE powder (BBcO5). X: Test performed

[0189] The powder stability study was conducted over 90 days and was stored at two different temperatures: 25°C and 40°C. The storage container consisted of a 20 mL glass vial with a metal cap and double-sealed parafilm closure system, minimizing contact between the semisolid formulation and the external environment. Various physical, chemical, and microbiological parameters were analyzed throughout the analysis period.

[0190] Before carrying out the battery of tests to assess the physical, chemical and microbiological stability of the formulation with 2% (w / w) of CIE powder, it was subjected to a mechanical stress test (centrifugation at 3500 RPMs for 30 minutes). It was found that, Regardless of storage temperature, the formulation remained stable over time after 90 days under two temperature conditions (25 9 C and 40°C).

[0191] From the photographic records taken throughout the stability study, it can be concluded that, macroscopically, there were no morphological changes, namely color and homogeneity (Figure 19). The odor of the CIE powder, woody and slightly woody, remained intense and constant throughout the analysis period.

[0192] At the same time, the color of the CIE powder formulation was analyzed more objectively using the colorimeter, which allows obtaining information regarding the luminosity (L*) parameters and chromatic coordinates a* and b*. It was therefore confirmed that no considerable differences were detected over the analysis time, regardless of the temperature (Figure 20).

[0193] The pH value of the formulation remained approximately constant for 90 days (Figure 21).

[0194] Regarding the rheological behavior of the formulation (Figure 22), similarly to what had already been verified for the base of the formulation, formulation 5 (BBcO5) maintained a non-Newtonian fluid behavior of the pseudoplastic type (rheofluidizing).

[0195] The behavior exhibited by BBcO5 cream over time (non-Newtonian fluid) is confirmed when applying the power law model. The behavior of formulation 5 (BBcO5) is the same as that described for the base formulation. The negative value of "n" indicates that the sample is more viscous at low shear rates, with the apparent viscosity decreasing as the shear rate increases (Table 16). A slight reduction in the consistency coefficient was observed, most significant at 40°C.

[0196] Table 16: Parameters of the power law model n=2.

[0197] The microbiological quality of formulation 5 (BBcO5 cream) was also analyzed after preparation and at time 90, after storage at different temperatures (25°C and 40°C). As described in ISO 21148:2017, the formulation was initially neutralized with a mixture of 3% (w / w) Tween80, 0.3% (w / w) lecithin, 3% (w / w) saponin, and 0.1% (w / w) histidine. The microbiological quality assessment was performed on samples of formulation 5 (BBcO5), either in sterile serum or enriched in TSB, to verify whether, in fact, there is microbiological growth during enrichment. When analyzing the count total microbiological, for the conditions used in the test, there was no growth (ISO 17516:2014). CORK POWDER EXTRACT Preparation of cork powder extracts

[0198] In a preferred embodiment, the extracts were initially obtained by treating the P1 cork powder fraction, corresponding to a finer and lighter cork powder obtained at a later stage of the cork industry production cycle, with heating to 40 °C with the following solvents: deionized water, 30% (v / v) ethanol, 50% (v / v) ethanol, 70% (v / v) ethanol, 96% (v / v) ethanol, and absolute ethanol. The choice of these solvents was based on the criteria of safety, compatibility with skin tissue, and sustainability. In each case, 5 g of cork powder (weighed) was mixed with 5 x 100 ml of solvent, and the mixtures were subjected to a stirring period of 5 x 5 h on a stir plate at 700 revolutions per minute (rpm). The extracts were filtered using a Büchner funnel with a glass fiber filter membrane (diameter 47 mm), and then concentrated at 40 °C under vacuum in a rotary evaporator.

[0199] After concentration, the extracts were collected in sample holders to determine their mass. The results are presented in the following table (Table 17).

[0200] Table 17 - Determination of extract mass

[0201] Based on the data obtained, al 9 extraction was the one that allowed the collection of the largest quantity of compounds existing in the cork powder. Among them, the extracts obtained from the mixtures at 70% (v / v) in EtOH and 50% (v / v) in EtOH were those that presented the greatest mass

[0202] As the number of extractions increased, there was a decrease in the mass of the extracts. From the 3 9 extraction, the variation in the mass of the extracts was not very pronounced, and therefore it was not considered profitable to carry out more than two extractions (Figure 23).

[0203] Cork powder extracts were then prepared under the same conditions at 40°C and at room temperature (RT, 20°C) using shorter extraction cycles, namely 3x1h / 2.5h, to establish the best extraction conditions based on the biological potential of the extracts and the sustainability of the extraction process. To predict the effect of temperature, extracting solvent, and number of extraction cycles, the amount of extract obtained was determined. (Figure 24A). Furthermore, the determination of absorbances at 280 and 350 nm served as an indicator of the extraction efficiency of phenolic compounds obtained using polar solvents (Figures 24B and 24C). Finally, the antioxidant activity of each extract was also determined by the DPPH radical scavenging capacity (Figure 25).

[0204] The scavenging activity of l,l-diphenyl-2-picrylhydrazyl (DPPH) free radical was evaluated according to a previously described method with some modifications. Absorbances were measured at 517 nm in a microplate reader (BioTek Synergy HT Instruments, Winooski, GU, USA). The percentage of inhibition activity was calculated according to the following formula: DPPH radical scavenging effect (%) = [1 - (Sample Abs - Sample Abs blank / Control Abs - Blank Abs)] x 100. Ascorbic acid was used as a positive control. The scavenging activity of the extract against the DPPH radical was expressed as the effective concentration at which the DPPH radical was scavenged by 50% (IC50). The IC50 value was obtained by interpolation of the linear regression analysis.

[0205] Overall, the extract masses obtained from extractions at room temperature and 40 °C were surprisingly similar. In both the 1h and 2.5h cycles, there was a greater increase in extract mass from the first to the second extraction than from the second to the third. A decrease in absorbance values was also observed, suggesting that the extraction of compounds of interest would decrease with the number of cycles, so that performing multiple extractions would not be advantageous.

[0206] Overall, all extracts showed promising antioxidant activity, with IC50 values mostly below 10 pg extract / mL (Figure 25). Comparison of the IC50 values obtained in successive extractions suggested that antioxidant potential decreased with the number of extractions, as evidenced by increasing IC50 values in almost all extracting solvents, regardless of extraction temperature. Interestingly, for almost all solvents, antioxidant activity appeared to be best in extracts prepared at room temperature and with a one-hour cycle.

[0207] Taking all these results into consideration, as well as the fact that room-temperature extract preparation is more sustainable and a single extraction cycle is easier to implement on an industrial scale, it was established that room-temperature extractions with a one-hour cycle constituted the optimized conditions. When analyzing the DPPH radical scavenging activity of extracts obtained at room temperature and with a one-hour extraction cycle, the extracts prepared with water, 30% (v / v), 70% (v / v), and 96% (v / v) ethanol revealed promising antioxidant activity and were selected for further studies.

[0208] Following optimized extraction conditions, new extracts were prepared using not only P1 cork powder but also P0 powder, a darker, coarser-grained powder obtained earlier in the cork industry's production cycle. These extracts were prepared at room temperature. environment for 1 hour with water and with hydroalcoholic solutions of 30% (v / v), 70% (v / v) and 96% (v / v) of EtOH. Cosmetic / pharmaceutical application of cork extracts

[0209] Tests to evaluate the potential antioxidant activity were carried out by the DPPH method in the extracts prepared from PO and Pl powders, according to the optimized conditions, namely 1h extraction at room temperature using water and the hydroalcoholic mixtures 30% (v / v) ethanol, 70% (v / v) ethanol and 96% (v / v) ethanol as extracting solvents (Table 18). In general, the extracts prepared with 30% (v / v) ethanol proved to be the most active, especially the extract obtained from Pl cork powder, which demonstrated the lowest IC50 value. Even so, all extracts showed promising antioxidant activity, with values below 10 pg of extract / mL, with the exception of the 96% (v / v) ethanol and aqueous extracts of P0 powder. Furthermore, P0 extracts showed lower antioxidant activity compared to Pl extracts.

[0210] Table 18 - Results of the DPPH radical scavenging activity of the P0 and Pl cork powder extracts. * Results expressed as mean ± standard error, n = 3. Ascorbic acid (positive control): ICso = 7.9 ± 0.4 pg / mL.

[0211] Next, the qualitative and quantitative composition of the P0 and Pl extracts was analyzed using HPLC. The analytical method was developed according to standards found in cork extracts reported in the literature. The method was validated according to international validation parameters. This technique allowed us to understand the effect of the extraction solvent on the extract profile, compare the composition of extracts obtained from different types of cork powder (P0 and Pl), and also attempt to relate them to the results obtained from antioxidant activity. The qualitative analysis of the chromatographic bands was based on the UV spectrum and mass spectrum (Table 19). The quantification of the compounds identified in the P0 and Pl cork powder extracts is presented in Table 20 and 21.

[0212] Many of the chromatographic bands did not match any of the studied standards, demonstrating the diversity of compounds present in the extracts. Although the profiles of the extracts obtained from the same powder are quite similar, some differences were observed, particularly in the intensity of some bands depending on the extraction solvent.

[0213] Table 19 - Ultraviolet spectrometry data and mass of identified peaks

[0214] In coarse cork powder extracts (PO), vescalagin was detected only in the extracts with the highest ethanol content, but at a concentration below the limit of quantification (LOQ) of the validated method for extract quantification. Castalagin was quantified at higher concentrations in hydroalcoholic extracts, being the main compound identified in the 70% EtOH and 96% EtOH ethanol extracts. The concentration of ellagic acid followed the same trend, increasing with the amount of ethanol in the extracting solvent, being highest in the 96% ethanol extract. In contrast, the concentration of gallic acid and latifolicin C was higher in the more polar extracts, i.e., those with the highest water content. The contents of the remaining compounds for all extraction solvents are presented in Table 20.

[0215] Table 20 - Quantification of compounds identified in cork powder extracts P0 Lq: Limit of quantification

[0216] In the fine cork powder extracts, P1, vescalagin was detected in all extracts, but quantified only in the 30% ethanol extract. In general, castalagin is the major compound in all extracts, except in the 96% ethanol extract, where ellagic acid is predominant. The concentration of gallic acid was higher in the more polar extracts, i.e., those with a higher water content. The concentration of ellagic acid was higher in the 96% EtOH extract. The concentration of gallic acid gallic acid and latifolicin C was lower in Pl extracts compared to PO extracts. The concentration of all identified compounds is presented in Table 21.

[0217] Table 21 - Quantification of compounds identified in cork powder extracts Pl. LQ: Limit of quantification

[0218] The improved antioxidant activity of the Pl cork powder extracts can be explained by the greater efficiency in extracting phenolic compounds. The extract prepared with 96% ethanol showed the lowest amounts of virtually all identified compounds (except ellagic acid), thus contributing to a higher IC50 value.

[0219] The P1 extract prepared with 30% ethanol demonstrated the best antioxidant activity, likely due to its higher ellagitannin content. Following the same logic, the P1 extract with 70% ethanol follows due to its higher content of castalagin, which is in fact the main compound identified in this extract. Next comes the P0 extract with 30% ethanol, possibly due to the concentrations of gallic and ellagic acids, and primarily latifolic acid C. In vitro evaluation of the toxicity of the extracts and their effect on inflammatory and antioxidant parameters

[0220] The toxicity of cork powder extracts prepared from P0 and Pl powders was evaluated under optimized conditions, namely extraction at room temperature using water and the hydroalcoholic mixtures 30% ethanol, 70% ethanol, and 96% ethanol, on representative cell lines of the skin (keratinocytes and human fibroblasts) and the immune system (macrophages). Cell viability was determined after exposing the cells to different concentrations (1000, 500, 250, 125, and 62.5 pg / mL) of each extract. The P0 and Pl powder extracts prepared with the hydroalcoholic mixtures 70% ethanol and 96% ethanol were discarded due to difficult dissolution in aqueous medium. From the results obtained with the remaining extracts, concentrations of 62.5 and 125 pg / mL were selected as they were the highest concentrations devoid of toxicity for subsequent tests (Figure 26).

[0221] The potential anti-inflammatory properties of cork dust extracts were subsequently evaluated. To this end, macrophages were exposed to cork extracts 30 minutes before incubation with the inflammatory stimulus lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria that activates innate immune cells through Toll-like receptor 4 (TLR4) binding. In turn, these cells, such as macrophages and neutrophils, synthesize pro-inflammatory mediators.

[0222] According to Figure 27, the P0-derived extract obtained with 30% ethanol and the Pl-derived extracts obtained with water or 30% ethanol inhibited LPS-induced nitrite levels in macrophages at both concentrations tested, with the inhibition being most pronounced at 125 pg / ml. As can be seen in Figure 27, the reduction in nitrite levels was not associated with reduced viability of cells exposed to LPS. These results demonstrate that these extracts have anti-inflammatory properties.

[0223] Subsequently, the effect of the extracts on the regulation of genes related to the inflammatory response and oxidative stress in macrophages (Figure 28) and keratinocytes (Figure 29), respectively, was investigated. A commercial cork cosmetic ingredient, DIAM Oleoactif® (INCI name "Cocos Nucifera Oil and Quercus Suber Bark Extract and Oak Root Extract"), which claims anti-inflammatory and antioxidant properties, was also studied for comparison. According to the results, all extracts increased the expression of the pro-inflammatory genes Nos2 (which encodes the inducible nitric oxide synthase (iNOS)), 11 IfJ (which encodes the interleukin-1 beta (IL-ip) protein), Tnfa (which encodes the tumor necrosis factor-alpha (TNF-a) protein), 116 (which encodes the IL-6 protein) and the anti-inflammatory gene 1110 (which encodes the IL-10 protein), in the absence of LPS (Figure 28).Surprisingly, under an inflammatory milieu (i.e., cells exposed to LPS), the aqueous Pl extract induced a significant reduction in Nos2 gene expression (which was not observed with Oleoactif), reinforcing its anti-inflammatory potential.

[0224] The expression of the antioxidant genes HM0X1 (which encodes the protein Heme Oxygenase-1 (HM0X1), TRX (which encodes the protein Thioredoxin (TRX)) and NQ01 (which encodes the protein NAD(P)H dehydrogenase quinone 1 (NQ.01)) was also evaluated, all proteins with relevant antioxidant activity (Figure 29).

[0225] While the Pl aqueous and P1-30%ETOH extracts induced a significant increase in the HM0X1 gene, only the Pl aqueous extract increased the expression of the TRX gene. However, it appears to reduce the expression of NQ.01, although without statistical significance (Figure 29). The results obtained suggest an antioxidant role of the Pl aqueous cork extract, an effect not observed for Oleoactif (Figure 29).

[0226] Since Pl extracts showed relevant anti-inflammatory activity by significantly reducing nitrite levels and Nos2 gene expression in macrophages exposed to LPS (1PC), and an antioxidant effect by significantly increasing the expression of the HM0X1 (1PC and 2PC) and TRX (1PC) genes in keratinocytes, Pl extracts were selected for subsequent experiments.

[0227] Thus, the effect of cork extracts (and Oleoactif) on iNOS protein levels in macrophages exposed to LPS was subsequently evaluated.

[0228] According to Figure 30, both PI extracts increased iNOS perse levels, but only the aqueous extract induced a significant decrease in LPS-induced iNOS protein levels, which is in agreement with the results obtained for gene expression (Figure 28, graph Nos2).

[0229] Since inflammation is closely linked to oxidative stress, the antioxidant potential of cork-derived extracts in oxidative stress triggered by an inflammatory stimulus was also evaluated. Thus, the effect of the extracts on HMOXl protein levels and mitochondrial reactive oxygen species (ROS), i.e., superoxide anion (O2), was determined in macrophages exposed to LPS (Figure 31A) or the mitochondrial stress-inducing compound rotenone (Figure 31B), respectively.

[0230] The results obtained showed that aqueous PI and 30% Pl-EtOH extracts increased the antioxidant enzyme HMOXl in immune cells, in the absence or presence of inflammatory stimulus (macrophages exposed to LPS) (Figure 31A). These results corroborate the ability of cork extracts to activate the antioxidant pathway in different cellular systems. Effect of cork-derived extracts on skin regeneration

[0231] The potential of PI extracts in skin regeneration was also investigated. To this end, the expression of the Collal and Itgbl genes (which encode type I collagen and the integrin beta 1 protein, respectively) (Figure 32A) and cell migratory capacity (using the scratch wound assay) (Figure 32B) were determined in fibroblasts exposed to cork extracts and the commercial compound Oleoactif. Surprisingly, the extracts significantly decreased the Collal gene and induced a slight decrease in the expression of the Itgbl gene (Figure 32A), and significantly reduced the migratory capacity of fibroblasts (Figure 32B). These results indicate that PI extracts inhibit cell migration. Anti-senescence effect of cork-derived extracts

[0232] The anti-senescence capacity of cork extracts was evaluated. Cellular senescence was induced in fibroblasts by etoposide, and the cells were subsequently exposed to aqueous extracts of PI and 30% EtOH and the commercial compound Oleoactif. As can be seen in Figure 33, both extracts (as well as Oleoactif) reduced the number of senescent cells, suggesting that they have anti-aging potential. Effect of extracts on skin irritation EpiSkin

[0233] The skin irritation potential of the aqueous PI extract was evaluated using a commercial reconstructed human epidermis model (Episkin™), according to the OECD Test 439 guideline. According to this skin irritation test, if the mean tissue viability is equal to or less than 50%, the sample is considered a skin irritant, which was not observed with the formulations containing the aqueous PI extract (Figure 34). Therefore, this extract demonstrates an adequate safety profile and does not induce skin irritation. Cosmetic / pharmaceutical composition comprising cork extract Pre-formulation and formulation of anti-aging cosmetic products based on cork extracts

[0234] Taking into account the bioactivity, yield of the obtained extract, sustainability of the process and transposition to industrial scale, the selected extraction conditions were as follows: • Extraction from Pl cork powder; • Extraction with aqueous solvent; • 1 cycle of LH extraction; • Room temperature.

[0235] The extracts were obtained from 100 ml of solvent to 2.5 g of powder, according to the following procedure: • Agitation in flask during cooking; • Filtration with l.2pm paper filter; • Evaporation in rotary evaporator; • Lyophilization.

[0236] Any lyophilization process may be applied to the product of the present disclosure. For example, in one embodiment the following protocol was applied: after rotary evaporation, the resulting residue is frozen at -80 °C, a vacuum (0.01-0.1 mbar) is applied, and then water is removed from the frozen residue by sublimation.

[0237] At this stage, production of the extract in bulk began in order to proceed with its physical and chemical characterization. Approximately 6 g of extract were obtained.

[0238] Characterization tests were carried out on the aqueous extract of Pl cork powder. This characterization included the determination of organoleptic characteristics (color, odor and appearance), pH determination, chemical profile, determination of heavy metals, assessment of microbiological quality and evaluation of the bioactivity of the extract obtained. Determination of organoleptic characteristics

[0239] The color of the extract was characterized using a colorimeter (ChromaMeter CR-400), recording the values of luminosity and chromatic coordinates (Table 22). Since this is a natural ingredient and it is not possible to control the color of each batch of extract obtained, this property is extremely important for assessing variability between batches and quality control, since the color obtained will directly influence the color of the formulation in which it will be incorporated.

[0240] Table 22: EAP1 L*a*b color parameters (n=3).

[0241] To determine the pH of the extract, a 10% (m / v) aqueous dispersion was prepared. The pH was measured in the filtrate with a Hl 2211 pH / ORP / °C meter (Hanna Instruments, Woonsocket, RI, USA). Measurements were performed in triplicate (Table 23).

[0242] Table 23: pH of aqueous extract n=3. Determination of heavy metal content

[0243] According to Regulation 1223 / 2009 on cosmetic products, the presence of heavy metals in ingredients can only be detected in trace amounts. Although the regulation does not specify legal limits, analysis of the present content is mandatory.

[0244] It was found that the heavy metal content is compatible with use in cosmetic products, and their presence did not result from intentional addition during the process of obtaining the extract (Table 24).

[0245] Table 24: Heavy metal content (mean ± SD) of the aqueous extract (ppm) n=3. Obtaining the chemical profile - Infrared Spectroscopy

[0246] Near-infrared (NIR) and mid-infrared (MIR) spectra were obtained for the aqueous extract (Figures 35 and 36). Obtaining the spectra at time zero allowed us to obtain a physical-chemical profile of the extract obtained. Although they alone are not sufficient to assess the extract's composition, they are of particular interest for assessing potential changes over time and for quality control. Pre-formulation studies Assessment of microbiological quality

[0247] The microbiological quality analysis protocol was prepared in accordance with ISO 21148:2017 and the European Pharmacopoeia 10. The sample was prepared by diluting 0.1 g of aqueous extract in 0.9 mL of saline. Serial dilutions were also prepared up to 10 -3. Aliquots of 100 pL of the original samples and dilutions were spread, in duplicate, on the surface of Tryptic Soy Agar (TSA) medium for bacterial isolation, and incubated for 24-48 hours at 30-35 ± 2 °C.

[0248] The microbiological quality of the aqueous extract was examined in accordance with ISO 21148:2017 and the European Pharmacopoeia 10, specifically the chapter "Microbiological examination of non-sterile products". The total microbiological count, for the conditions used in the test, was below the legal limits (2.00 x 10 3 UFC / g) established by ISO. Evaluation of bioactivity and total phenolic compounds

[0249] As demonstrated, the aqueous cork extract, obtained from the Pl powder by-product, presents antioxidant activity.

[0250] In order to characterize the new extract obtained, the antioxidant activity was evaluated through the DPPH method and the total phenolic content (TPC) was determined according to the Folin-Ciocalteu method, with some modifications (Table 25). The TPC was calculated using a calibration curve plotted with gallic acid and expressed in mg of gallic acid equivalent per g of dry extract (mg GAE / g of extract).

[0251] Table 25: Total phenolic content and DPPH scavenging activity (IC5o) of the aqueous extract at time zero (T0, start of the stability study). n=3 Solubility test

[0252] The solubility of the aqueous extract was evaluated according to OPPTS 830.7840 guidelines to evaluate potential solvents for inclusion in cosmetic product formulations. Three solvents were selected: water, glycerin, and 1,3-butylene glycol. The extract was soluble in water and glycerin at a ratio of 1g / L and insoluble in butylene glycol. Stability Study of Aqueous Cork Extract

[0253] The stability study was carried out over 90 days and was stored at two temperatures: 25 and 40 °C. The tests were carried out as mentioned in Table 26. Table 26: Summary of aqueous extract stability protocol. X: test performed

[0254] The storage container used was a glass jar, protected with aluminum foil on the outside to minimize the potential effect of light.

[0255] Through the photographic records taken throughout the stability study, it is possible to conclude that, macroscopically, there were no organoleptic changes, particularly in color (Figure 37).

[0256] At the same time, a more objective analysis of the color of the aqueous extract was performed using a colorimeter, which allows obtaining information regarding the parameters luminosity (L*) and chromatic coordinates a* and b* (Figure 38). Therefore, it was confirmed that no considerable differences were detected over the analysis time, regardless of the temperature. The aqueous extract presented a woody odor, which remained unchanged throughout the analysis period.

[0257] The pH remained relatively stable, between pH values of 4.5 and 5.3 throughout the analysis period and independent of temperature (Figure 39).

[0258] Near-infrared (NIR) and mid-infrared (MIR) spectra were also obtained for the aqueous extract (Figure 40) initially and throughout the stability test (TO, T15, T30, T60, and T90) for both temperatures (25 and 40°C). They allowed verifying that the chemical composition of the extract remained unchanged over time, since no new bands appeared or existing bands disappeared in the obtained spectra. The overlap of the spectra also reveals its stability (Figure 40).

[0259] Microbiological quality was assessed at time 0 of the test and at 90 days, in samples stored at temperatures of 25°C and 40°C. The test was performed inside a horizontal laminar flow chamber to reduce possible risks of sample contamination. The sample aqueous Pl extract (1:10 dilution) was prepared and properly homogenized. Then, serial dilutions (10 1 , IO-2 and IO -3 ), in duplicate. The total count of total aerobic microorganisms was 5.00 x 10 2 UFC / g, 48h after the start of the analysis.

[0260] The procedure was repeated 90 days after the stability study, for both the sample stored at 25 °C and at 40 °C. In these cases, the aerobic mesophilic microorganism count was 3.00 x 10 2 UFC / g e 5.00 x 10 2 UFC / g, respectively. In all situations, it is lower than the limits established in ISO 21148:2017 (2 xlO 3 CFU / g).

[0261] The antioxidant activity of the aqueous extract was also evaluated throughout the stability test using the DPPH assay. IC5o values were evaluated at 0, 15, 30, 60, and 90 days for the PI aqueous extract stored at 25 and 40 °C. Ascorbic acid was used as a positive control (IC5o = 7.9 ± 0.4 pg / mL) (Figure 41).

[0262] The aqueous extract was moderately stable at temperatures of 25 and 40 °C, in relation to the DPPH scavenging effect.

[0263] The IC50 of the extract at 25 °C ranged from 5.79 ± 1.07 pg / mL at time 0 to 9.92 ± l.20 pg / mL at time 90 days. For the extract stored at 40 °C, the IC50 ranged from 5.79 ± 1.07 pg / mL at time 0 to 9.64 ± l.20 pg / mL at time 90 days.

[0264] The antioxidant activity of the extract remained present at the end of the 90 days of storage at both temperatures.

[0265] Total phenolic compounds were also evaluated at time 90 (Table 27). Table 27: Total phenolic content and DPPH scavenging activity (IC50) of the aqueous extract. Comparison between t0 and t90 (n=3).

[0266] The stability of the extract was also evaluated in a 3.5% (w / w) glycerol solution. The IC50 values at 25°C ranged from 9.17 ± 0.34 pg / mL (TO) to 4.91 ± 1.30 pg / mL (T90). The results showed that the incorporation of the extract in this solution is quite stable, with only a slight variation in the DPPH uptake capacity being observed over the 90 days (Figure 42).

[0267] The pH values remained constant, at values close to pH 5, throughout the test (Figure 43).

[0268] In order to evaluate the stability of the aqueous extract at different pH values, two buffer solutions were prepared at pH 5 and 7. The extract proved to be more stable in the pH 5 buffer solution (Figure 44). Base Formulation for Extract Incorporation Test (EAP1)

[0269] The base formulation was selected in order to study the stability of the developed ingredient, when incorporated into cosmetic / pharmaceutical product formulations, with a face cream being selected for incorporation of the aqueous extract (EAP1).

[0270] In order to verify the physical-chemical stability over time, the respective formulation stability assessment protocol was also developed. Formulation Stability with Aqueous Cork Extract

[0271] Regarding the developed formulations, the physical stability protocol was implemented, namely with the evaluation of rheological behavior.

[0272] From the formulations tested, the CR002 face cream formulation was selected to incorporate 1% (w / w) lyophilized extract, as it was more pleasant to the touch and had a better visual appearance (Table 28).

[0273] Table 28: Composition of the CR002 formulation and respective preparation procedure. [00274 Over the 3 months of the trial the following tests were performed (Table 29).

[0275] Table 29: Summary of the stability protocol of the formulation with 1% (w / w) incorporated extract. X: Test performed Initial characterization of the formulation

[0276] After selecting the base formulation (face cream reference CR002), lyophilized cork extract was incorporated into it at a concentration of 1% (w / w).

[0277] Before beginning the stability test, the base formulation and the extract formulation were subjected to a mechanical stress test to preliminarily assess their stability (centrifugation at 3500 rpm for 30 minutes). Since no phase separation was observed, the formulation is considered stable.

[0278] In order to characterize the formulation with incorporated extract, its spreadability was evaluated using a texturometer.

[0279] The ease with which the conical probe moves down the cream reflects the product's spreadability. As soon as the probe comes into contact with the sample, the force increases steadily until a maximum value is reached. The probe then reverses direction and withdraws from the sample. In this case, the measured force is negative because the formulation adheres to the probe, resisting its movement out of the sample container (Figure 45). It can be concluded that storage temperature does not contribute to significant differences in the spreadability of the aqueous cork extract formulation analyzed. Evaluation of the organoleptic characteristics of the formulation during the stability test

[0280] The color of the formulations was evaluated using a colorimeter (Tables 30 and 31). Due to the characteristic brown color of the extract, it gave a light brown hue to the basic formulation, which was white in color.

[0281] Table 30: Color parameters of the base formulation. Table 31: Color parameters of the base formulation with aqueous extract.

[0282] The color of the formulations stored at 25 and 40°C was analyzed over 90 days to determine whether this parameter remained stable (Figure 46). In fact, it was concluded that both the luminosity and the chromatic coordinates A and B of the formulation with the extract remained practically unchanged throughout the analysis period, regardless of the temperature. pH assessment

[0283] The pH assessment over the 3 months of the stability test (Figure 47) showed that the pH decreased slightly over time (minimum pH value 4.3). However, it remains at values considered suitable for use on the skin. Assessment of microbiological quality

[0284] The microbiological quality of the formulation under study was evaluated initially and after 90 days of storage at 25°C and 40°C.

[0285] The microbiological evaluation of cosmetic products is mandatory for batch release and must be carried out in accordance with ISO 21148:2017 and 17516:2014 standards, and Pharmacopoeia 10.

[0286] The manufacture of cosmetic products must follow Good Manufacturing Practices, which must mitigate contamination. The microbiological quality of a cosmetic product should not depend solely on its preservative system. Therefore, assessing microbiological quality requires the use of a neutralizing system, which inactivates the preservative, allowing the growth of microorganisms, if present in the formulation.

[0287] Thus, samples of the formulations stored at 25 °C and 40 °C were prepared under the following conditions: • Sterile serum sample (1:10) • Sample in neutralized serum (1:10) • Sample in sterile serum enriched in TSB (1:10) • Sample in neutralized serum enriched in TSB (1:10)

[0288] As controls, sterile serum, neutralized serum and TSB were also incubated to ensure that they were not a source of sample contamination.

[0289] The neutralizing system was selected according to the instructions of the manufacturer of the preservative system used. After inoculation and incubation of the plates, respectively without dilution and dilutions at 10 1 , IO -2 and IO -3 , it is verified that the microbiological count is within the legal limits established by ISO (2xlO 3 CFU / g). Rheological behavior Flow

[0290] The runoff assessment was carried out initially and after 90 days (Figure 48).

[0291] Apparent viscosity is a variable that characterizes a system rheologically. Assessing this parameter helps determine whether a product has the appropriate consistency or fluidity and can indicate adequate stability, providing an indication of the product's behavior over time.

[0292] The behavior exhibited by CR002 cream over time (non-Newtonian fluid) is confirmed when applying the power law model. The value of the flow index (n) indicates that the viscosity decreases as the shear rate increases (Table 32).

[0293] Table 32: Power Law Model Parameters.

[0294] The samples remained stable and without phase separation, although slight changes in rheological behavior were observed.

[0295] It can also be stated that both the aqueous cork extract and the formulation in which it was incorporated remained stable during the 3-month stability test. The extract demonstrated its antioxidant activity throughout the stability test, thus allowing us to conclude about the viability of this ingredient as an active ingredient in cosmetic products. The term "comprises" or "comprising" when used herein is intended to indicate the presence of the features, elements, integers, steps, and components mentioned, but does not preclude the presence or addition of one or more other features, elements, integers, steps, and components, or groups thereof. The following claims define additional embodiments of the present description.

Claims

CLAIMS 1. Cork powder for use in cosmetics or medicine comprising cork particles with a particle size of less than 100 pm, and in which the color has L* values between 45-55; a* values between 6-10 and b* values between 12-16, measured by the CIELAB system.

2. Cork powder according to the preceding claim comprising cork particles with a particle size of less than 63 pm; preferably ranging from 16-63 pm; more preferably 20-40 pm. Cork powder according to any one of the preceding claims, wherein the cork powder is subjected to 180°C for at least 30 minutes; preferably 30 minutes. Cork powder according to any one of the preceding claims, wherein the oil absorption capacity of the cork powder is at least 4 times greater than that of kaolin.

5. Method for obtaining cork powder according to any one of claims 1-4, comprising the following steps: providing cork powder comprising cork particles with a particle size of less than 100 pm; preferably less than 63 pm; adding water and stirring for at least 15 minutes in order to obtain a dispersion of cork powder; filtering the dispersion of cork powder through a porous glass filter in order to obtain clean cork powder; drying the clean cork powder at at least 50°C for at least 1 h, in order to obtain dry cork powder; heating the dry cork powder at 180°C for at least 30 minutes.

6. Cork powder according to any one of claims 1-4 or method according to claim 5, wherein the cork powder is selected from: grinding powder, cleaning powder, powder from particle size separations, powder from finishing agglomerate panels, powder from finishing cork stoppers and discs; powder from finishing natural cork stoppers, or a mixture thereof; preferably powder from particle size separations.

7. Aqueous extract of cork powder for cosmetic or medical use comprising: castalagin, in a concentration greater than 10 pg / mg (dry extract); 46 gallic acid, in a concentration greater than 2 pg / mg (dry extract); ellagic acid, in a concentration greater than 4 pg / mg (dry extract). Extract according to the preceding claim, wherein the extract is obtained from the cork powder according to any one of the preceding claims 1-4.

9. Extract according to any one of claims 7-8, comprising: castalagin, in a concentration ranging from 10-60 pg / mg (dry extract), preferably from 10-50 pg / mg (dry extract); gallic acid, in a concentration ranging from 2-20 pg / mg (dry extract), preferably from 2-15 pg / mg (dry extract); ellagic acid, in a concentration ranging from 4-30 pg / mg (dry extract), preferably from 4-20 pg / mg (dry extract).

10. Cosmetic or pharmaceutical composition comprising the cork powder according to any one of claims 1-4 or the aqueous extract of cork powder according to any one of the preceding claims 7-9.

11. Composition according to the previous claim comprising 1-7% (m / m) of cork powder; preferably 4-6% (m / m) of cork powder; more preferably 5% (m / m).

12. Composition according to the preceding claim comprising 0.1-4% (m / m) of cork powder; preferably 1-3% (m / m) of cork powder; more preferably 2% (m / m). Composition according to any one of the preceding claims 11-12 further comprising titanium dioxide; preferably 1-15% (w / w) titanium dioxide; more preferably 8-12% (w / w).

14. Composition according to the previous claim comprising: 1-7% (m / m) cork powder; preferably 5% (m / m); 1-15% (w / w) titanium dioxide; preferably 10% (w / w). Composition according to any one of the preceding claims 11-14 further comprising: a mixture of glyceryl monostearate, cetostearyl alcohol, cetyl palmitate and coco-glycerides; preferably 2-8% (w / w); ethoxylated cetostearyl alcohol (ceteareth-12); preferably 0.1-3% (w / w); 47 medium-chain triglycerides; preferably 4-12% (w / w); cetearyl isononanoate; preferably 4-12% (w / w).

16. Composition according to any one of the preceding claims 11-15 wherein the sun protection factor is at least 40; preferably between 40-80.

17. Composition according to any one of the preceding claims 11-16 further comprising: 2-8% (w / w) of a mixture of polyglyceryl-6 esters of olive oil, sodium stearoyl lactate, cetostearyl alcohol; 4-12% (m / m) dicapryl ether; 1-4% (m / m) squalene; 1-3% (m / m) caprylic / capric acid triglycerides; 1-4% (m / m) stearic acid; 1-3% (w / w) sweet almond oil; 0.1-0.5 (m / m) Xanthan gum; 1-3 %(w / w) Propanediol; 2-8% (w / w) Glycerin; 0.1-2% (w / w) of a mixture of phenoxyethanol, benzyl alcohol, ethylhexylglycerin and tocopherol.

18. Composition according to the previous claim in which the color of the cosmetic composition has an L* between 40-54; an a* between 4-8 and a b* between 10-17, measured by the CIELAB system.

19. Composition according to the preceding claim 10 comprising 0.1-2% (m / m) of aqueous extract of cork powder; preferably 0.1-1% (m / m); more preferably 0.5-1% (m / m).

20. Composition according to the previous claim, further comprising: 4-6% (w / w) of a mixture of olive oil polyglyceryl-6 esters, sodium stearoyl lactate, cetostearyl alcohol; 10-20% (w / w) of triglycerides of capric acid and / or caprylic acid; 1-3% (w / w) cetostearyl alcohol; 0.1-0.2 (m / m) xanthan gum; 1-3% / (w / w) propylene glycol; 0.1-2% (w / w) of a mixture of phenoxyethanol, benzyl alcohol, ethylhexylglycerin and tocopherol. 48 21. Composition according to any one of the preceding claims 19-20, wherein the color of the composition has an L* between 48-55; an a* between 2 and 7; and a b* between 15-20, measured by the CIELAB system.

22. Composition according to any one of the preceding claims 10-21, wherein the composition is selected from a list consisting of: solution, serum, cream, lotion, gel, hydrogel, oleogel, oil, soap, shampoo, dry shampoo, spray, ointment, paste, mousse or body bath gel; preferably cream.

23. Use of the cork powder according to any one of claims 1-4 or of the composition according to any one of the preceding compositions 11-18 as an oil-absorbing agent.

24. Use of the cork powder according to any one of claims 1-4 or of the composition according to any one of the preceding compositions 11-18 as a pigment; preferably as a cosmetic pigment.

25. Use of the cork powder according to any one of claims 1-4 or of the composition according to any one of the previous compositions 11-18 as a sun protection factor enhancing agent, preferably wherein the enhancement factor is at least 2 times; preferably the enhancement factor ranges between 2-3 times.

26. Use of the cork powder according to any one of claims 1-4 or of the composition according to any one of the previous compositions 11-18 as an ultraviolet radiation blocking agent.

27. Cork powder according to any one of claims 1-4 or composition according to any one of the preceding compositions 12-21 for use in medicine or veterinary medicine.

28. Cork powder according to any one of claims 1-4 or composition according to any one of the preceding compositions 11-18 for use in the prevention and treatment of skin diseases; preferably skin diseases caused by exposure to the sun.

29. Use of the extract according to any one of claims 7-9 or the composition according to any one of claims 19-22 as a preventive or delaying agent for skin aging.

30. Use of the extract according to any one of claims 7-9 or the composition according to any one of claims 19-22 as an anti-senescent agent.

31. Use of the extract according to any one of claims 7-9 or the composition according to any one of claims 19-22 as an antioxidant agent.

32. Extract according to any one of claims 7-9 or composition according to any one of claims 19-22 for use in medicine; preferably for use in the prevention or treatment of inflammation.

33. Extract according to any one of claims 7-9 or composition according to any one of claims 19-22 for use as an anti-inflammatory; preferably a topical anti-inflammatory.

34. Method of obtaining the extract as described in any one of the preceding claims 7-9 comprising the following steps: providing cork powder comprising cork particles with a particle size of less than 0.5 mm; adding to the cork powder an aqueous solvent in a mass ratio of cork powder:aqueous solvent ranging from 1:20 to 1:80, and stirring for at least 30 minutes to obtain a mixture comprising the aqueous extract of cork powder; filtering the mixture comprising the aqueous extract of cork powder to obtain an aqueous extract of cork powder.

35. Method according to the preceding claim comprising an additional step of drying and lyophilizing the aqueous extract of cork powder so as to obtain a lyophilized aqueous extract of cork powder.

36. Method according to any one of the preceding claims 34-35, wherein the particle size of the cork particles is less than 250 pm; preferably between 63-200 pm; more preferably 100-200 pm.

37. Method according to any one of the preceding claims 34-36, wherein the mass ratio of cork powder: aqueous solvent ranges from 1:30 to 1:50; preferably, the mass ratio of cork powder: aqueous solvent is 1:

40.

38. Method according to any one of the preceding claims 34-37 wherein the agitation step of the aqueous solvent and the cork powder ranges from 500-1000 rpm; preferably 700 rpm.

39. The method of any one of the preceding claims 34-38 wherein the aqueous solvent is water.

40. The method of any one of claims 34-39 wherein the method is carried out at room temperature.

41. Method according to any one of the preceding claims 34-40 wherein the stirring time of the aqueous solvent and the cork powder ranges from 30 - 120 minutes; preferably 60 minutes.

42. Method according to any one of the preceding claims 34-41, wherein the filtration step is carried out with a glass fiber filter membrane; preferably with a diameter of 47 mm and a pore size of 1.2 pm.

43. Method according to any one of the preceding claims 34-42 wherein the extraction step is carried out in a single extraction cycle.

44. Extract according to any one of the preceding claims 7-9 or method according to any one of claims 34-43, wherein the cork powder used is selected from: grinding powder, cleaning powder, powder from particle size separations, powder from finishing agglomerate panels, powder from finishing cork stoppers and discs; powder from finishing natural cork stoppers, or a mixture thereof; preferably powder from particle size separations. SUBSTITUTE SHEET (RULE 26) 3 / 37 a ( ) 4 / 37 SUBSTITUTE SHEET (RULE 26) 5 / 37 SUBSTITUTE SHEET (RULE 26) 6 / 37 SUBSTITUTE SHEET (RULE 26) 7 / 37 SUBSTITUTE SHEET (RULE 26) 9 / 37 None SUBSTITUTE SHEET (RULE 26) 10 / 37 Fig. 12 SUBSTITUTE SHEET (RULE 26) 11 / 37 12 / 37 SUBSTITUTE SHEET (RULE 26) 13 / 37 14 / 37 Fig. 17 SUBSTITUTE SHEET (RULE 26) 15 / 37 SUBSTITUTE SHEET (RULE 26) 16 / 37 SUBSTITUTE SHEET (RULE 26) 17 / 37 18 / 37 19 / 37 SUBSTITUTE SHEET (RULE 26) 20 / 37 SUBSTITUTE SHEET (RULE 26) 21 / 37 RAW Fig. 26 A SUBSTITUTE SHEET (RULE 26) 22 / 37 23 / 37 Fig.27 SUBSTITUTE SHEET (RULE 26) 24 / 37 Fig. 30 SUBSTITUTE SHEET (RULE 26) 25 / 37 Fig. 31 Fig. 32 SUBSTITUTE SHEET (RULE 26) 26 / 37 27 / 37 Fig. 36 SUBSTITUTE SHEET (RULE 26) 28 / 37 29 / 37 30 / 37 Fig. 42 SUBSTITUTE SHEET (RULE 26) 33 / 37 Fig. 43 SUBSTITUTE SHEET (RULE 26) 34 / 37 SUBSTITUTE SHEET (RULE 26) 35 / 37 Fig. 45 SUBSTITUTE SHEET (RULE 26) 36 / 37 37 / 37 Fig. 48 SUBSTITUTE SHEET (RULE 26)