BRPI1004637A2 - cosmetic products containing extracts and microalgal components and processes for their production - Google Patents

Abstract

COSMETIC PRODUCTS CONTAINING MICROALGAL EXTRACTS AND COMPONENTS AND PROCESS FOR THE PRODUCTION OF THE SAME. The present invention deals with the development of cosmetic formulations with added microalgal products and extracts. Several species of microorganisms, among them the cyanobacterium Spirulina plotensis, have been studied for their possible biological activities in the areas of health and nutrition. However, many of its properties can be used for the production of cosmetic products, such as its antioxidant, anti-aging, healing, anti-acne, oil-reducing or cellulite properties, among others. In addition, several of its products can be used as enhancers of rheological and sensory characteristics, providing products with greater spreadability, consistency, less residual grease sensation, shine, drying speed, among others.

Description

COSMETIC PRODUCTS CONTAINING EXTRACTS AND MICROALGAL COMPONENTS AND PROCESS FOR THE SAME PRODUCTION

Field of the Invention

The present invention is concerned with the development of cosmetic formulations with added microalgal components and extracts in order to bring improved characteristics to such products as biological activity (antioxidant power, anti-aging, healing, anti-acne, oil-reducing, anti-acne). cellulite, among others) and improved rheological and sensory properties (higher spreadability, consistency, less residual grease sensation, luster, drying rate, etc.).

Invention History

Cosmetic formulations are known and have been developed for centuries. Base formulations are easy to find and manipulate, and the major differential is the additives used in each formulation, giving it the best characteristics and properties. Therefore the extraction of active ingredients from plants, fruit, roots, etc. is widely studied. Most of the substances used to preserve cosmetic products are synthetic substances such as BHA (2 and 3-tert-butyl-4-hydroxyanisol) and BHT (di-tert-butyl methyl phenol), which has been studied over the last few years. , and demonstrated possible toxicity when there is long term exposure. In addition, as rheological enhancers, finite source mineral materials such as kaolin and talc are currently used. Thus, the present invention aims at the development of novel cosmetic products with specifically microalgal bioactive source molecules capable of imparting similar or better properties to the cosmetic product.

Multifunctional cosmetics are products that have functions beyond their basic function. This makes its use more comprehensive and improves its performance, being preferably used by the consumer. In this context, the product presented in this invention has in addition to moisturizing property, strong antioxidant property. Creams with antioxidant properties often use chemicals as preservatives and antioxidants (such as BHA - 2 and 3-tert-butyl-4-hydroxyanisol and BHT - di-tert-butyl methyl phenol), whose safety has been questioned.

BHA is a carcinogenic and skin irritant, and its use should be limited to the values suggested as safe by specific laws.

BHT has low acute toxicity. Although it causes skin irritation, according to studies with pigs, the compound does not appear to cause irritation upon ingestion of permitted food use levels. In acute exposure studies in mice, BHT has shown toxic effects on the lungs, including hyperplasia, hypertrophy, and a general disruption of cellular constituents.

TBHQ (t-butylhydroquinone) has low acute toxicity and although in pig studies has shown slight skin irritation, it is not considered an irritant compound and should be ingested at the permitted levels in food. Some evidence indicates that TBHQ has a low genotoxic potential and may be slightly mutagenic at high doses.

Interest in natural antioxidants began in the 1980s due to evidence of harmful effects caused by high doses of BHT, BHA and T-BQ (t-butylhydroquinone) as liver overweight and endoplasmic reticulum proliferation, among others. Consequently, emphasis has been placed on identifying and purifying new compounds with antioxidant activity from natural sources that may act alone or in synergy with other additives as an alternative to prevent oxidative deterioration of cosmetics and replace the use of synthetic antioxidants.

State of art

Algae and microalgae and their products are widely used in the development of cosmetic products such as creams, gels, shampoos, etc. Heterogeneous polysaccharides, called mucilages, are extracted from brown (Feoficeas) and red (Rodoficeas) algae. When in contact with water, this mucilage forms a colloidal but non-adhesive system, much appreciated by the cosmetic industry. Among these compounds, the most used are alginate, agar and carrageenans. Other species, such as Fucus vesiculosus (a red algae with emollient properties) have the ability to accumulate iodine, and are used in anticellulitic treatments.

Macrohystis pyrifera biomass powder has sebaceous secretion regulating properties. Rhodymenia palmaria has sweating properties. Blood Delesseria and Punnatifada Undalaria act on the skin toning, moisturizing and anti-wrinkle action. Dunaliella saline is a source of pro-vitamin A (beta carotene), and its extracts stimulate cell regeneration at the epidermis or dermis level. In addition, it appears to stimulate melanin production and has antioxidant power. Plocanium coccineum is a source of red pigments that can replace synthetic dyes, reducing the risks of toxicity, even though they have less stability than the former.

Microalgal polysaccharides are important metabolites, representing 40 to 90% of the organic compounds produced by these microorganisms. They have a wide spectrum of compositions and different molecular masses. In general, microalgae polysaccharides can be divided into three major groups: reserve, structural and extracellular.

The main polysaccharides are the reserve polysaccharides, usually composed of homogeneous glucans such as starch and their variations in cyanobacteria and rhodophytes.

Structural polysaccharides, present mainly in cell walls, can vary greatly in composition depending on the taxonomic group, and include xylans, mananas, rhamnanas, and various glycoproteins and are used as an important criterion in defining microalgae phyla, classes, and orders. Extracellular polysaccharides have a very variable composition and can form homogeneous compounds containing fucans, mananas, arabinogalactans, and heteropolysaccharides containing five or more types of monosaccharides.

Many of the known polysaccharides have been studied for their biological activity. The large number of fractions with different molecular weights and different compositions, produced by microalgae species not yet well studied, shows the possibility of the discovery of many other bioactive compounds.

Fucanas are polysaccharides rich in fucose, and have a broad spectrum of biological activity, including anticoagulant, anti-inflammatory, immunostimulatory, anti-viral, anti-tumor and anti-metastasis effect. The main sources are brown algae (Feoficea Class), particularly from the Laminaria and Fucales orders, which produce highly sulphated fucanas known as fucoidan. Many microalgae species may also produce bioactivated fucans, but tests in this area are still scarce. Crypfomonas obovata, Tetrapyrenoidosa cryptomonas (Cryptophyceae), Duostra thalassiosira (BaciIIariophyceae) and Staurastrum orbiculare (Zygnematophyceae) are examples of freshwater microalgae producing polysaccharides rich in fucose.

Arabinans, arabinogalactans and ramnogalacturans are often associated with a class of compounds known as pectins, characterized by the presence of arabins, galactose and galacturonic acid. These compounds are very common in higher plants and plants used in traditional medicine. Many algae species, especially those phylogenetically close to higher plants, have pectins in the cell wall. In addition, other green microalgae exhibit cell wall and pectin-like extracellular polysaccharides, such as glycoproteins (feroporins) that form the typical envelope of Clamidomonadales Bars and Volvocales. The immunostimulatory activity of Chlorello sp. (ChlorococcaIes) is directly related to the presence of a polysaccharide rich in arabinose and galactose.

Considering the described aspects, it is evident that the production of bioactive polysaccharides by microalgae should be better explored. Bioprospecting is an indispensable tool for the discovery of new substances of human interest.

Microalgae production of EPS (exopolysaccharides) often occurs under high or low pH and high light conditions. A large production of EPS leads to a decrease in biomass harvesting efficiency, making filtration and centrifugation difficult. On the other hand, the presence of a certain amount of EPS in the medium seems to facilitate harvesting, causing flocculation.

Spirulina sp. secretes sulfated exopolysaccharides through capsule formation and gradient release into the culture medium. The promising applications of Spirulina EPS are related to its biological activity and have been investigated as antitumor, antioxidant, antibiotic, antiparasitic activity, etc.

There are no products or studies exactly similar to those proposed in this invention. Some creams use marine extracts in their composition, as described below.

An example is La Prairie's "Advanced Marine Biology Cream", which promises to slow the aging process through advanced antioxidant marine actives that appear to firm the skin through hyaluronic acid, and to have a "Protective Complex", It fights aging with rice ceramides, seaweed extract and green algae that help stimulate collagen production. According to the manufacturer, moisturizing actives (seaweed, corn extracts and other moisturizing agents) are used in its composition. It promises to protect the skin through "marine actives for antioxidant protection with mariponic technology-grown seaweed extracts and marine perejil that give the skin an antioxidant capability of unprecedented defenses". Additional information can be found at http://www.laprairie.com

Another example is Multi Vegetal's "Seaweed and Herb Face Cream", which is said to be a nourishing cream for facial skin with nutrient-rich plant extracts and natural antioxidants that act as free radical suppressants that cause cell death and skin aging. According to the references, it contains Seaweed - Fucus vesiculosus (antioxidant and natural iodine source), Babosa -Aloe sp. (nourishing and soothing) Chamomile - Chamomilla recutita (soothing, tonic, refreshing and anti-inflammatory), Ginkgo-Ginkgo biloba (anti-inflammatory, antioxidant and blood circulation stimulant) and Herb Mate -llex paraguariensis (powerful moisturizer and natural antioxidant). Available at: http://www.multivegetal.com/creme-facial-com-algas-e-ervas.php

However, in this type of product, the extracts used are macroalgae ("seaweed"), not microalgae. Seaweed (macroalgae) has slower and more delicate growth than microalgae, which can easily be adapted to a large-scale industrial production process.

In the work "Disease of a cream for massage with extract of Cuban spirulina, by González et al, 2005, published in the Cuban Journal of Pharmacology, Institute of Pharmacy and Food, University of La Habana", the preparation of a formulation is reported. of cream with Spirulina hydroalcoholic extract, unlike the object of this invention, wherein an aqueous extract is used. Moreover, González et al 2005 do not describe the use of EPS produced by microalgae. In addition, the purpose of using the product is different, as our work was to develop a moisturizing skin cream for continuous use, while in the work of González et al, 2005, the goal is to use it in massages, and may even be removed after use, not in continuous use.

Some patents already report the use of microalgae and their compounds for the production of cosmetic products. However, none of the patents found to date use neither the same microorganisms nor the same processes to obtain a product with antioxidant capacity and improved rheological and sensory properties as disclosed in the present patent.

W02010054322A1 entitled "Cosmetic Compositions Compising Microalgal Components" utilizes whole microalgal cells with a certain carotid content, and contains at least 10% oil and is free of any other oil. These creams should be applied by compresses, which last between 1 and 3 hours, not for topical or continuous use. In this work, the use of EPS of the tested microorganisms to improve rheological or sensory characteristics is not mentioned.

Already in patent W003000222A2 "Cosmetic Preparation With Anti-wrinkle action" is used an aqueous extract of microalgal biomass from which a peptide is purified, which appears to combat skin aging.

In WO 2007/110511 "Cosmetic active ingredient composed of arginine ferrulate and microalgae extract and its issues" the objective is to increase the activity of skin proteosomes to increase thioredoxin production and to reduce the production of glycolyzed proteins. This is done by obtaining a hydroglycolic extract capable of inducing such reactions. It is obtained by extracting biomass from microalgae Chlorella, Scenedesmus and Spirulina, and a green coffee extract.

FR276499A1 "Cosmetic Product for the Protection of Epidermis of the Effects of Air Pollution" aims to produce a cosmetic product capable of protecting the skin against pollution using extracts of marine microorganisms of the genus Phormidium. It uses extracts up to 0.1% by weight in the final product, and is based on maintaining or increasing the amount of intracellular ATP in epidermal cells. FR2785909A1 "Microalgae Extract Containing Antiradical Substances and Polysaccharides Sulfates and Composition 1'incorporant" discloses a process for obtaining microalgal materials of antioxidant activity. This activity is due to the action of the enzyme superoxide dismutase obtained by forced metabolic stress. The species used are Porphyridium cruenfum and Rodophytic species.

Patent MD3781F1 "Proceeded to obtain the preparation of antioxidant thermostable cyanobacterial biomass Spirulina platensis" discloses a production process using Gromov culture medium added with tartaric acid and imidazole. The obtained cells are disrupted by freezing / thawing and the recovery process of the active ingredients includes the use of superoxide dismutase.

US7651690 "Purified component of blue-green algae and method of use" describes the use of microalgae such as Aphanizomenon and Spirulina for the production of selectins, describing their isolation, purification and use.

Brief Description of the Invention

The present invention deals with the development of cosmetic formulation containing aqueous microalgae biomass extract, which has antioxidant properties, and the polysaccharide obtained from the same microalgae, for the improvement of rheological and sensory aspects of the cosmetic product.

Citation of Figures (if any)

Figure I illustrates the growth curve of 5. platensis over 15 days. Growth parameters were calculated: Pmax = 1.44g.L "1; Pmax = 0.42g.L'1.d'1, and Mmax = 0.43 day1.

Figure 2 illustrates the antioxidant potential test by the chemiluminescence methodology, which can be used to calculate the IC50 of the aqueous biomass extract. Figure 3 illustrates the antioxidant potential test by the chemiluminescence methodology, which can be used to calculate the Spirulina EPS IC50.

Figure 4 shows curves obtained by rheological analysis of the creams. At the top are flow curves of the base cream (a), the cream with 0.5% EPS (b); the 2.5% EPS cream of Spirulina (c); and cream with 5.0% EPS of Spirulina (d). These behaviors show that the addition of higher EPS values (2.5 and 5%) negatively affects the product thixotropy, greatly increasing or decreasing the hysteresis areas (which can be seen in the figure as the areas between the upward curves). and descendants). The most appropriate behavior is compatible with that shown in Figure (b), showing that the concentration of 0.5% EPS showed the best result.

Figure 5 shows curves obtained by rheological analysis of the creams. At the top, they are strain sweeping curves of base cream (a), cream with 0.5% EPS (b); the 2.5% EPS cream of Spirulina (c); and cream with 5.0% EPS of Spirulina (d). These behaviors show that the determination of higher EPS values (2.5 and 5%) acts negatively on the behavior of the formulation's G 'and G "modules. For Spirulina Base cream and with 0.5% EPS, This behavior was practically constant, as shown by the almost completely horizontal curves, while for the other two formulations, containing 2.5 and 5.0% EPS, this behavior was not verified, with distortions in the behavior of the curves. Appropriate analysis is consistent with that shown in Figure (b), showing that the concentration of 0.5% EPS showed the best result.

Figure 6 shows curves obtained by rheological analysis of the creams. At the top, they are frequency sweep curves, base cream (a), cream with 0.5% EPS (b); the 2.5% EPS cream of Spirulina (c); and cream with 5.0% EPS of Spirulina (d). These behaviors show that the addition of higher EPS values (2.5 and 5%) negatively affect the behavior of the formulation's G 'and G "modules. For Spirulina Base Cream and 0.5% EPS, module G "was practically constant and horizontal, and the module G ', despite presenting a slightly increasing (inclined) behavior, presented it more smoothly than the others. For the other two formulations, containing 2.5 and 5.0% EPS, this behavior was not verified, with distortions in the behaviors of the two curves. Thus, again, the most appropriate behavior is consistent with that shown in Figure (b), showing that the concentration of 0.5% EPS showed the best result.

Figure 7 shows curves obtained by rheological analysis of the creams. In this case, they are curves of the test called Creep and Recovery, the base cream (a), the cream with 0.5% EPS (b); the 2.5% EPS cream of Spirulina (c); and cream with 5.0% EPS of Spirulina (d). Base Cream and 0.5% EPS cream showed virtually the same recovery, while for the other two formulations, the recoveries were lower, indicating that these two creams should be more fluid than the first two, causing problems with its application and use, may run down the skin, and may also have a less creamy appearance.

Figure 8 shows water activity curves of each cream under each condition tested in the accelerated stability test. It can be noted that the water activity remained practically constant after 90 days of analysis for all conditions tested for Spirulina Cream (green marked points) while the base creams had a large loss of water activity (marked points). in red), which can be explained by the water retention capacity of the EPS.

Figure 9 shows the averages of spreadability found for each condition tested in the accelerated stability test. It can be noted that at higher temperatures (Incubator) the cream loses free water and dryness, which directly interferes with its spreadability value (both Base Cream and Spirulina Cream had low values of spreadability in this condition, when compared to other conditions. ). It is also important to note that for all conditions Spirulina Cream had the best spreadability value compared to the Base Cream formulation, showing that EPS improved not only sensory characteristics but also rheological characteristics became better.

Figure 10 shows the result of the sensory evaluation performed with the Base Creams and Spirulina. The grades could range from 1 to 9. It can be noted that for the Spirulina Cream, object of the present invention, all ratings were better.

Detailed Description of the Invention

The process of obtaining cream with differentiated antioxidant and sensory properties begins with the cultivation of microalgae. Given the cultivation time, biomass is collected by a physical separation process, preferably gravitational, such as filtration and centrifugation. Subsequent drying of the biomass is performed at 60 ° C, but may vary between room temperature and 100 ° C. After drying the biomass is ground. The biomass is diluted and homogenized in polar solvent (10 to 50% w / v), preferably water. The mixture is centrifuged for biomass separation and the extract (supernatant) is used.

The supernatant, which is separated from the microalgae culture, is concentrated (up to 4 times its initial volume) in rotary evaporator and ethanol precipitated (up to 4 volumes of ethanol are used for each concentrated extract volume). The mixture is left in a freezer (-5 to -20 ° C) for better decantation of exopolysaccharide (EPS). After centrifugation, the remaining salts are removed by dialysis preferably with a 16kDa dialysis membrane (but may range from 7 to 22kDa). The purified polysaccharide is then lyophilized for later application. The base cream used is an oil / water emulsion containing: Ethoxylated Ketostearyl Alcohol, Ketostearyl Alcohol, Silicone Fluid (Dimethicone Copolyol), Cetyl Palmitate, Isopropyl Myristate, Propylparaben, Methylparabenoyl Propylene Glycol, Ethoxylated Lanolin 50%, and distilled water. The composition of the cream base is not restricted to that described; Other base creams may be used to which the microaiga extract and its EPS will be added. 0.5 to 5% of the microalgal polysaccharide is added to the base cream, and from 1 to 10 mL of a 5-40% aqueous extraction of the biomass. The samples are homogenized and the pH adjusted between 5 and 7 with acid / base solution.

There is no need to use the whole microaiga cell, which would not dissolve in the cream, creating sensory problems and even smelly. These problems are avoided by the formulation proposed in this invention while maintaining the antioxidant properties of the microaiga. Another advantage observed in the proposed formulation is the elimination of the need to use preservatives of chemical origin (synthetic molecules that are commonly used for this purpose, and whose safety has been questioned lately, such as BHT and BHA). In addition, the added polysaccharides may provide the cream with improved and unique sensory properties that would not be obtained with the use of other molecules / ingredients.

The following examples show the use of EPS and biomass extract from the Spirulino platensis microaiga. However, as previously mentioned, the cosmetic applications of microalgae aqueous extract and EPS are not restricted to this species.

Example 1: Biomass Production and EPS Obtainment and Power Analysis

Antioxidant of Biomass and EPS The Spirulina platensis strain was grown in 6 liter Erlenmeyers (4L useful volume) at 30 ° C in Zarrouk culture medium. Lighting was provided by daylight fluorescent lamps providing a total of 3500 lux. The light period used was 12:12 h (light / dark). Agitation was performed with air injection at 0.2-0.3 vvm. The initial concentration of S. plafensis toi of 0.23g.L-1 and biomass growth was monitored by dry weight.

Samples were taken daily and centrifuged in a Mach Legend Sorvall 1.6 R centrifuge (Sorvall, Germany) at 16800xg for 15 min. The cells were washed with water and dried at 60 ° C to constant weight while the cell free medium was used for total sugar analysis by the phenol sulfuric method (Dubois, M., Gilles, KA, Hamilton, JK, Rebers, PA, Smith, E., 1956. Colorimetric method for determination of sugars and related substances (Anal. Chem. 28, 350-356.), Estimating EPS production.

Dry biomass was analyzed for chlorophylls, carbohydrates, proteins, lipids and ashes. The pigments were extracted with 90% acetone and quantification followed the equations suggested by (Strickland, J.D.H., Parsons, T. A practical handbook of seawater analysis Pigment analysis. Bull. Fish. Res. Bd. Canada 167, 311. 1968). Lipids were determined by extraction with 1: 1 methanol: chloroform followed by a liquid-liquid extraction with hexane (Sydney, E.B., et al. Potential carbon dioxide fixation by industrially important microalgae. Bioresour. Technol. 20 2010). Ash was quantified by the AOAC 941.12 method, while the phenol sulfuric method was used to determine total carbohydrates and the Lowry method (Lowry, OH, NJ Rosebrough, AL Farr and RJ Randall1 1951. Protein measurement with the Folin-Phenol J. Biol. Chem. 193: 265-275) for protein determination. Phenolic compounds 25 were quantified by the Folin-Ciocalteu method (Costa, DA, Chaves, MH, Silva, WCS, Costi, CHS. Chemical constituents, total phenols and antioxidant activity of Sterculia striata St. Hil et Naudin. Acta Amaz. Vol. 40 No. 1 Manaus, Mar. 2010), and the antioxidant potential was determined by two distinct methodologies: the 2,2-diphenyl-1-picrylhydraz 30 (DPPH) assay system and chemiluminescence testing using 10% ( w / v) of aqueous biomass extract. The S. platensis culture was vacuum filtered on filter paper (Whatman 1). The filtrate was concentrated to% of the original volume of vacuum rotary evaporator below 50 ° C. The concentrate was then dialyzed on a 12-14 KDa dialysis membrane. For the precipitation of EPS, four volumes of absolute ethanol were added to the obtained concentrate under vigorous stirring. This mixture was left overnight at 4 ° C. After centrifugation at 16800xg for 10 min, the supernatant was discarded. The EPS was lyophilized and saved for subsequent applications.

Extracellular polysaccharide concentration was determined according to the classical method of Dubois et al. (1956) using glucose solution as a reference standard.

Analysis of the EPS composition was performed on a Shimadzu brand High Performance Liquid Chromatrography (HPLC) equipped with a refractive index detector. The column used was Aminex HPX87H, with the analysis conditions: 5mm H 2 SO 4, 60 ° C, 0.6 mL / min.

The growth curve obtained for S. platensis cyanobacteria can be seen in figure 1.

The general composition of Spirulina platensis biomass in terms of proteins, sugars, pigments, lipids and ashes is shown in table 1.

Table 1: Centesimal composition of S. platensis biomass.

<table> table see original document page 15 </column> </row> <table>

Phenolic compounds tests were performed on biomass and polysaccharides, using an initial dissolution of 10% (v / w). Spirulina platensis contains many types of pigments, including chlorophyll-a, xanthophyll, beta-carotene, echinenone, myxoxanthophyll, zeaxanthin, canthaxanthin, diatoxanthin, 3'-hydroxyechinenone, beta-cryptoxanthin, and cophanocyanocyanin and phycobilocyanine. All listed pigments have at least one characteristic phenolic region, which may play a role as an antioxidant. The content of total tenol compounds was 150.39 mg per 100 g of biomass. The amount of chlorophylls a and b and carotenoids are shown in Table 2.

Table 2: Chlorophyll and carotenoids quantification of S. platensis biomass.

<table> table see original document page 16 </column> </row> <table>

The antioxidant potential of Spirulina platensis biomass was quantified by chemiluminescence method. Bar graphs shown in figure 4 indicate the signal measured by the equipment, proportional to the free radical content. Thus, samples with superior antioxidant power have smaller response areas by this method.

The solvent used in the dilutions was phosphate buffer, pH 7.4. A control was made with distilled water.

It is clearly seen from Figure 2 that the biomass sample had an almost 100% free radical inhibiting ability at a concentration of approximately 2.0 mg / mL. Thus, the IC50 of the sample can be calculated, which equals 0.962 mg / ml.

By comparison, BHT (di-tert-butyl methylphenol) has an IC 50 of 12.5 pg / mL and BHA (tert-butyl-4-hydroxyanisol) of 2.97 µg / mL.

The production of EPS by Spirulina platensis was followed for 14 days, reaching O ^Gl-1, which means 0.183gEPS / gbiomass (18.3%). The theoretical balance for Spirulina platensis can be up to 51%, especially when grown under stress conditions (temperature, lighting and / or pH, for example). Freeze-dried non-dialyzed EPS looked very similar to kaolin. Both have low density, light appearance and low density.

The monosaccharide composition of the EPS produced is shown in Table 3. Hexoses are the main constituents of EPS (mannose, glucose and galactose), but a large amount of pentose was also observed (rhamnose and fucose).

Table 3: Protein and carbohydrate content in EPS and their monosaccharide composition.

<table> table see original document page 17 </column> </row> <table>

The amount of phenolic compounds in the EPS was 7.44 mg (in 100 g EPS sample). To quantify the antioxidant potential of EPS the Iuminol-dependent chemiluminescence method was used (Figure 3). The IC50 of EPS was 2.888 mg / ml. This value is three times greater than that of biomass extract, which means that the biomass extract tested is three times more efficient against free radicals than EPS itself.

Example 2: Rheological analysis of creams plus different EPS concentrations

Four sets of creams were prepared using the formulation already described. Spirulina EPS was added to three of these creams at increasing concentrations of 0.5%, 2.5% and 5.0%. One control was used without addition of EPS. The creams were kept in opaque jars at 4 ° C. Four hours prior to analysis, the creams were left at room temperature until thermal equilibrium.

Rheological analysis was performed on a Mod Brooktield RV III rheometer using a cone-plate system, Rheocalc 3.0 Software. and a CP 52 spindle. The analysis was performed in triplicate using 0.4 g of the sample. All creams tested (with or without the addition of Spirulina EPS) showed non-Newtonian, pseudoplastic and viscoelastic and thixotropic behavior. The most appropriate hysteresis area was achieved with Spirulino platensis EPS 0.5%. This same composition presented the best apparent viscosity, indicating better spreadability in relation to the other creams.

Frequency and voltage scans were performed to observe the behavior of the G 'and G modules of each sample. Base cream and 0.5% EPS cream showed the most constant pattern, which indicates better emulsion stability (distorted in other creams, probably by the large amount of solid particles). Figures 4 to 7 illustrate the tests performed with creams with different concentrations of Spirulina EPS.

Example 3: Stability Analysis

One kilogram of base cream was prepared, which was divided into two parts, in which 0.5% (w / w) EPS of Spirulina platensis and 2ml of aqueous biomass extract (10% v / w) were added. )The. Each part was divided into 5 120g plastic jars each containing 90-100g of cream. The vials were distributed to assess response to different conditions. One jar of base cream and one base cream containing S. platensis biomass extract and EPS were left under the following conditions: room temperature, refrigerator (5 ° C), oven (37 ° C), shaker (120rpm, 27 ° C) and illuminated shelf (3500lux, 27 ° C).

The stability of the test creams was evaluated for 90 days (accelerated stability). Spirulina cream had better water retention capacity (Figure 8), indicating greater moisturizing power.

Both creams showed negative variations when incubated at elevated temperatures, as there was water loss, changing the overall appearance of the cream and reducing its spreadability (Figure 9). For all other conditions, Spirulina cream had better spreadability values than base cream.

Example 4: Sensory Analysis of Cream containing Aauous Extract of

biomass and EPS

Sensory analysis was performed on a group of 35 volunteer adults. Everyone was informed about the content of the proof and the risks inherent in it. The tests were performed only after signing the agreement and adhesion agreement.

Both creams evaluated had the same basic composition. The difference between the so-called "Base Cream" of the "Spirulina Cream" was the addition of 0.5% EPS of Spirulina platensis microalgae and 4mL / kg of 10% aqueous biomass extract.

Two kits were prepared, each containing the samples and supporting materials (spatulas, water, towel and paper napkins). The samples were randomly numbered so that the volunteer could not establish a logical relationship between them.

Four initial skin hydration measurements were made prior to application of the creams using the Corneometer CM825 device. The regions were demarcated on the forearm of the panel members, with points at the square vertices measuring 3.5 cm χ 3.5 cm (12.25 cm 2) for application. A mold was used because the measurement of skin hydration must be done exactly in the regions where the creams were applied. The group members were then invited to start the evaluation and to answer the questionnaire.

For application a small portion of the evaluated creams was removed from the container with a spatula, avoiding contamination through the contact of the panel member fingers with the remaining cream in the jar. Once placed on the skin, the cream was spread by hand in 15-20 circular motions until completely absorbed.

The questionnaire was based on an affective analysis. The following parameters were chosen: appearance, consistency, drying speed, brightness, spreadability, tackiness, residual grease sensation and overall impression. In addition, two questions were added regarding the cream's buying attitude. The first question was about the buying attitude based on the characteristics of the cream evaluated, and the second question about the influence of the presence of a natural antioxidant extract on the cream formulation.

Spirulina cream had a better rating for its consistency, drying speed, glossiness, viscosity spreadability, and residual grease sensation. The overall impression was also better for Spirulina Cream. The buying potential of Spirulina cream was much higher than Base Cream. The summary of the evaluation of each aspect of each cream can be seen in figure 10.

The moisturizing power of the Spirulina-containing cream was higher than the base cream, showing that in addition to improving sensory properties, the addition of EPS provides greater skin hydration.

When considering the possible existence of a natural antioxidant in the creams, the buying intent by the panel members increased.

Example 4: Irritability Analysis

The methodology chosen to evaluate potential irritating creams was the Draize Method, with modifications. A group of 10 male 120 day old albino guinea pigs (Cav / a porcellus) were used. Before starting the experiment, they were "left in the Vivarium for three weeks for acclimatization. When they reached 142 days of age, the test was performed. Two areas of the back of the animals were scraped, 8cmx5cm, 24 hours prior to application. 0.5 g of each cream (one base cream and one base cream containing 10% aqueous biomass extract and 0.5% EPS) was applied to each area of all animals. It is necessary to cover the test sites with surgical gauze as recommended in some methodologies as the cream was completely and rapidly absorbed into the animals' skin.The animals were divided into two groups of five: one group received base cream and the other , the cream of spirulina.

Application sites were visually observed immediately after application and after 24 and 72h. Edema, erythema and bedsores, as well as reversibility of possible reactions after seven days were observed.

Both EPS and base cream did not cause any skin irritation on guinea pigs. Thus, the IIC (skin irritability index) was zero.

Claims (27)

1. COSMETIC PRODUCTS CONTAINING MICROALGAL EXTRACTS AND COMPONENTS AND PROCESS FOR THE SAME PRODUCTION, characterized by the use of microalgal exopolysaccharides and microalgal biomass extracts. 2. COSMETIC PRODUCTS CONTAINING MICROALGAL EXTRACTS AND COMPONENTS AND PROCESS FOR PRODUCTION THEREOF, according to claim 1, characterized by the addition of component (s), including microalgal biomass extracts and microalgal exopolysaccharides in the product formulation. 3. COSMETIC PRODUCTS CONTAINING EXTRACTS AND MICROALGAL COMPONENTS AND PROCESS FOR PRODUCTION THEREOF, according to claims 1 and 2, characterized by the action of microalgal exopolysaccharide as an enhancer of rheology, stability and sensory properties of these products. 4. COSMETIC PRODUCTS CONTAINING MICROALGAL EXTRACTS AND COMPONENTS AND PROCESS FOR PRODUCTION THEREOF, according to claims 1 and 2, characterized in that the microalgal biomass extract is a source of antioxidant substances. COSMETIC PRODUCTS CONTAINING EXTRACTS AND MICROALGAL COMPONENTS AND THE PROCESS FOR PRODUCING THEME, using a base cream for the addition of products, including microalgal extracts, according to claim 1, characterized by the use of the following formulation: surfactant / emulsifier, including ethoxylated stearyl alcohol in a concentration ranging from 2 to 4%; an emollient emulsifier including stearyl alcohol in a concentration ranging from 4 to 8%; Silicone fluid (dimethicone copolyol) = 0.5 to 3%; emollients, including cetyl palmitate in a concentration ranging from 1 to 5% and Isopropyl myristate in a concentration ranging from 2 to 6%; preservative and antimicrobials including Propylparaben in a concentration ranging from 0.001 to 0.1% and Methylparaben in a concentration ranging from 0.01 to 1%; a humectant and solubilizer including propylene glycol in a concentration ranging from 1 to 5%; a stabilizing and conditioning agent including 40 to 60% ethoxylated lanolin at a concentration ranging from 0.05 to 5% and water. 6. COSMETIC PRODUCTS CONTAINING EXTRACTS AND MICROALGAL COMPONENTS AND THE PROCESS FOR PRODUCING THEMS, according to claims 1 and 5, characterized by the use of any other cosmetic formulation for topical use on human body parts, including hair, skin, mucous membranes and nails . Process for the production of COSMETIC PRODUCTS CONTAINING EXTRACTS AND MICROALGAL COMPONENTS according to Claim 1, characterized in that it comprises the following steps: - Cultivation of microalgae; - Physical separation of biomass and supernatant containing EPS; - Drying of the remaining biomass; - Crushing of dry biomass; - Dilution and homogenization of biomass in polar solvent; - Polar solvent evaporation if no aqueous solvent is used (optional); - Centrifugation of the resulting material to separate the supernatant containing antioxidant solution from the biomass; - Concentration of the first extracted supernatant containing the EPS; - Precipitation of EPS with addition of polar solvent; - Decantation of precipitated EPS; - Dialysis of the resulting EPS for salt removal (optional); - Preparation of EPS base cream, antioxidant solution and base formulation. COSMETIC PRODUCTS CONTAINING MICROALGAL EXTRACTS AND COMPONENTS AND THE PROCESS FOR PRODUCING THEMSELVES according to claims 1 and 7, characterized in that the microalgae cultivation step uses microalgae, comprising Amphidinium sp. and other members of the class Inophyfa Chlorachnion sp. and other organisms of the Chlorachniopyhto class, Botryococcus sp., Chiamydomonas sp., Chlorella sp., Chloroehytrium sp., Cholococcum sp., Chorieystis sp., Coeeobotrys sp. , Desmodesmus sp., Dicyococcus sp., Dunaliella sp., Haematoeoceu sp., Mierospora sp., Pediastrum sp., Pseudochlorella sp., Seenedesmus sp., Tetradesmus sp., Tetraselmis sp., Tetraspora sp. sp., and other members of the Chlorophyeeae class, Mieromonas sp. and other members of the Prasinophyeeae class, Mieromonas sp., and other members of the Prasinophyeeae sp. class, Aetinastrum sp., Desmoeoceus sp., Muriella sp., Nannoclhoris sp., Ooeystis sp., Cladophoropsis sp. members of the Ulvophyeeae class, Chroomonas sp., and other members of the Cryptophyeeae class, Anabaena sp, Aphanizomenon sp., Aphanoeaspa sp., Arthrospira sp., Calothrix sp., Crinalium sp., Fiseherella sp., Fremyella sp. , Limnothrix sp., Lyngbya sp., Mieroeoleus sp., Microeystis sp., Nodularia sp., Nostoe sp., Oscillatoria sp., Spirulina sp., Synechococcus sp. and other members of the Cyanobateria Euglena sp. and other members of the Euglenophyeeae class, Cyanophora sp., and other members of the Glaueophyta division, Isoehrysis sp., Pavlova sp. and other members of the Haptophyta division, CyeIoteIIa sp., Phaeodaetylum sp., Skeletonema sp., Thalassiosira sp. and other members of the Bacillariophyceaef Chromulina sp. and other members of the Chrysophyeeae class, Nannochloropsis sp., Phaeobotrys sp., Heterosigma sp., Botrydium sp., Heteroeoeeus sp., Xanthonema sp., and other members of the Heterokontophyta division, Cyanidium sp., Dixoniella sp. Porphyra sp., Porphyridium sp., Rhodospora sp. and other members of the Rhodophyeeae class; Choleoehaete sp., Arthrodesmus sp., Cosmarium sp., Desmidim sp., Euastrum sp., Spirogyra sp., Zygnema sp. and other members of the Charophyta division. Process according to Claim 7, characterized in that the microalgae cultivation step is carried out at a constant aeration of 0.2 to 0.3 vvm over a period of 10 to 30 days at a temperature of 20 ° C. at 35 ° C, using lighting cycles including natural and artificial light, with addition of carbon sources including sodium bicarbonate (from -14 to 18 gL-1) and minerals including (in gL-1) : K 2 HPO 4 (from 0.1 to 1.5); NaNO3 (from 1 to 4.5); K 2 SO 4 (from 0.5 to 2.0); NaCl (from 0.5 to 2.0); MgSO 4 JH 2 O (from 0.1 to 1.0); CaCl2 (from 0.01 to 0.08); EDTA (from 0.02 to 0.1) and 1.0 mL.L- 'of solutions A5 and B6. Solution A5 (g.L- '): H3BO3 (from 2 to 3); MnCMH 2 O (from 1 to -3); ZnSO 4 JH 2 O (from 0.1 to 0.5); CuCO4.5H2 O (from 0.05 to 0.1); MnO3 (from 0.01 to 0.05). Solution B6 (gL-i): NH4VO3 (from 20 to 25); KCr (SO 4) 2.12H 2 O (from 150 to -210 ° C); NiSO4.6H20 (from 30 to 50); Na2W04.2H20 (from 15 to 20); TiSO4.H2SO4.8H-2 (55 to 70); C0 (NO3) 2.60H2 (from 35 to 50). Process according to Claim 7, characterized in that the physical separation of the biomass may be carried out by means of a gravitational separation process comprising centrifugation and filtration. Process according to Claims 7 and 10, characterized in that the centrifugation step for separating the biomass and the EPS-containing solution is carried out in a range of 7000 to 15000 rpm. Process according to Claim 7, 10, characterized in that the biomass drying step is carried out at a temperature of from 20 ° to 100 ° C over a period of 1 to 15 days. Process according to Claim 7, characterized in that the step of diluting the biomass after grinding is carried out with a polar solvent including water, ethanol, methanol and chloroform at a concentration of 5 to 20% w / v. . Process according to Claims 7 and 13, characterized in that it does not require the polar solvent evaporation step, since it is an aqueous extract. Process according to Claim 7, characterized in that the step of centrifuging the polar solvent diluted biomass is carried out at 1000 to 10000 rpm and separating the extracted biomass and the extract containing antioxidants. Process according to Claims 7 and 11, 25 characterized in that the concentration of the EPS-containing solution is carried out by evaporation of 50 to 90% of the liquid phase. Process according to Claims 7, 11 and 16, characterized in that the EPS is precipitated from the concentrated solution by the addition of double to quadruple polar solvent relative to the volume of solution comprising ethanol. Process according to Claims 7 and 11, characterized in that the settling step of the EPS after precipitation is carried out at a temperature of -4 ° C to 25 ° C for a period of 4 to 24 ° C. hours Process according to Claims 7, 11 and 18, characterized in that it contains a centrifugation step of 5000 to 10000 rpm for a period of 10 to 30 minutes after decanting the EPS for purification of the EPS. Process according to any one of the preceding claims, characterized in that the step for making the final cream comprises a mixture between EPS and the antioxidants purified with a base cream. Process according to one of the preceding claims, characterized in that the EPS is provided in powder form through a drying process including lyophilization. Cosmetic Products Containing Microalgal Products and Extracts according to any of the preceding claims, characterized in that from 1 to 10 ml antioxidant-containing extract is added per 50g of final cream. Cosmetic Products Containing Microalgal Products and Extracts according to Claims 1, 5, 7, 20 and 22, characterized in that an exopolysaccharide of dry microalgal origin is added to the base cream. Cosmetic Products Containing Microalgal Products and Extracts according to Claims 1, 5, 7, 20, 22 and 23, characterized in that the drying of the exopolysaccharides is carried out by lyophilization. Cosmetic Products Containing Microalgaisi Products and Extracts according to claims 1, 5, 7, 20, 22 and 23, characterized in that the drying of the exopolysaccharides is performed by dialysis. Cosmetic Products Containing Microalgal Products and Extracts according to Claims 1, 5, 7, 20 and 22, characterized in that it adds to the base cream an exopolysaccharide of wet, non-dialyzed and non-lyophilized origin. Cosmetic Products Containing Microalgal Products and Extracts according to Claim 1, characterized in that 0.1 to 10% (w / w) of exopolysaccharide is added to the cream formulation.