RO136021A2 - Cosmetic cream composition with nourishing, moisturizing and calming effect - Google Patents

Abstract

The invention relates to a cosmetic cream composition with nourishing, moisturizing and calming effect. According to the invention, the composition consists of 20 parts of black cumin oil, 5 parts of oil-in-water emulsifier obtained from olive oil, 2 parts of co-emulsifier, such as a plant-based fatty alcohol from coconut oil or palm oil, 68.58 parts of aqueous re-extract of truffle/pure water, 3 parts of plant-based glycerin, 0.1 parts of Boswellia serrata as powder, 0.6 parts of preservative, 0.45 parts of lavender essential oil, 0.15 parts of mountain arnica extract and 0.12 parts of vitamin E, the parts being expressed by mass.

Description

STATE OFFICE FOR INVENTIONS AND TRADEMARKS Patent application _No. ck 2^21 Λ ΔΟ Γ i deposit ...Xl..'J3.~.202l..

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COSMETIC CREAM COMPOSITION WITH NUTRITIVE EFFECT,

MOISTURIZING AND SOOTHING

The invention refers to a cosmetic cream composition with a nourishing, moisturizing and soothing effect for revitalizing the skin, with use in the medical and cosmetic field.

skin care is an indispensable part of the gear called health, but no less important are the products we use.

An organ of great aesthetic value, the skin provides protection for the body, being, at the same time, a huge gateway to the body. For this reason, cosmetic products are responsible for: cleaning without destroying the protective barrier of the skin, nourishment, source of nutrients and exfoliation by removing dead cells from the surface layer, protection against ultraviolet rays.

Because most of the ingredients in cosmetics are absorbed by the skin, natural products come with a gentle, skin-friendly alternative with great responsibility towards the production process, testing and the environment.

Skin creams are used to improve the health of the skin and to create suppleness, softness and flexibility.

The present paper refers to a cosmetic product in the form of a cream intended for the normalization of skin maintenance processes, as well as the method of obtaining it.

In order to improve the formulations with topical skin applications, many researchers use substances rich in proteins, vitamins, these being considered ingredients capable of binding water and hydrating the skin, being also biocompatible with the skin tissue.

From the patent literature in the field of medical and dermato-cosmetic products based on boswellia as such or in association with various other pharmaceutical acceptable substances, the following can be mentioned:

Patent KR20040078498A, entitled "Cosmetic composition for improving skin elasticity, containing Boswellia extracts as an active ingredient" describes a cosmetic composition for increasing skin elasticity, comprising Boswellia extract as an active ingredient, thus inhibiting elastase activity and glycosaminoglycan decomposition. Therefore, skin wrinkles can be removed and skin elasticity can be improved.

The cosmetic composition for improving skin elasticity comprises 0.001 to 15% by weight of Boswellia extract as an active ingredient, wherein the Boswellia is selected from Boswellia serrata, Boswellia frereana and Boswellia carterii; and the composition

cleanser containing surfactant, oil, powder foundation, emulsion foundation, wax foundation and spray.

Boswellic acid according to Patent KR20040078498A was purchased from Doosan Biotech (Korea) and used. In addition, Boswellia serrata was purchased from Korea Herbal Medicine and used to prepare the extracts.

Patent RO125277, with the title "Ultra-active anti-acne cream", describes a face cream used as part of an anti-acne treatment. The cream according to the invention contains, as active principles, 0.1.....0.3% 5-chloro-2-(2,4-diclophenoxy)-phenol, 0.5.....5% anti-acne propylene glycol plant extract from Ocinum basilicum, Phaseolits vulgaris, Hedera helix, respectively, extract in dipropylene glycol of Boswellia serrata, 0.1...10% lactic acid, 1...5% zinc oxide, respectively, sulfur, in a cream base with known excipients .

The invention patent CN107375145B, with the title "Cosmetic matrix, method of preparation and application thereof, cream for eye contour and method of preparation thereof", describes a cosmetic matrix, which can be used in the preparation of cosmetic products for skin care and a contour cream eyes containing the cosmetic base, which can effectively reduce melanin in the skin around the eyes, having the effect of brightening the eyes and effectively removing dark circles.

The cosmetic matrix is mainly prepared from the following raw materials in mass percentages: 10-20% tea extract with Boswellia serrata extract, 5-10% citrus peel extract, 10-20% date seeds, 5-10% acetyl tetrapeptide, 3-6% of a blend of wild soybean protein, hydrolyzed rice protein, and superoxide dismutase. 3-6% Boligonal cuspidatum root extract, 1-3% ascorbyl tetraisopalmitate. 1-2% hydrogenated PEG-40, castor oil and water.

Moreover, based on the technical scheme provided by the invention, the eye contour cream is prepared mainly from the following raw materials in mass percentages: 20-30% cosmetic matrix, 2-3% hydrogenated castor oil PEG-60, 1-2 % cetostearyl polyether alcohol, 1-2% behenyl alcohol, 2-3% shea butter, 3-4% dioctyl carbonate, 3-4% glycerol tri(ethyl hexanoate), 0.5-1% tocopherol acetate, 8-10% glycerol, 5-7% butanediol, 0.5-0.8% carbomer, 0.3-0.6% triethanolamine and water.

Patent US7883726, entitled “Skin care compositions with botanical seed oils”, describes an improved topical skin care composition (lotion) based on botanical seed oils, prepared according to a cold-pressed method, for the treatment for dry skin, anti-wrinkle, mild eczema, psoriasis and as an anti-aging balm

The composition of the skin care lotion has a range of about 99.9% (% w/w) to 98% (% w/w) of the topical skin care composition and a range of about 0.1 (% w/ w) up to 2% (% w/w) of botanical seed oil blends such as red raspberry seed oil and a plurality of botanical oils selected from the group consisting of the following seed oils: pumpkin seed oil, chardonnay grape seed, carrot seed oil, blueberry seed oil, cranberry seed oil, pomegranate seed oil, black cumin seed oil and black raspberry seed oil.

New botanical oils are natural emollients. They prevent drying and protect the skin, acting as a barrier and healing agent, as well as a soothing and softening agent for the skin.

Products with a smoothing effect on the epidermis are known, in which the improvement of the skin microrelief is done by increasing hydration, with the help of ingredients such as polyols (glycerin, propylene glycol, sorbitol), alpha hydroxy acids or urea, which have the ability to bind water to the surface of the epidermis , of the type of mucopolysaccharides (hyaluronic acid), with the ability to accumulate water deep in the skin, or ingredients that restore the protective lipid barrier, preventing water loss, such as mineral or vegetable oils (rich in ceramides, essential fatty acids, phytosterols), or selected ceramides.

The present invention refers to a new cream composition, which has the advantage of having in its composition the aqueous re-extract from brown dessert truffle cake, natural negrilica (black cumin) oil, Boswellia serrata powder, vegetable glycerin, lavender essential oil and vitamin E, in well-established ratios, which gives the cream a good physical-chemical and biological stability and revitalization (regeneration) capacity by restoring the skin's elasticity, hydration and the supply of skin-specific components: proteins, natural fats and vitamins. By constantly applying this cosmetic cream composition with a nourishing, moisturizing and soothing effect, wrinkles are reduced, and a good revitalizing action is obtained.

Technical problem

The objective technical problem proposed to be solved by this invention application consists of:

- establishing the components and their association ratios for the preparation of a cream rich in proteins and vitamins for skin hydration and revitalization.

- the replacement of some chemical synthesis components in the formulas of U/A (oil-in-water emulsions) or A/U (water-in-oil emulsions) cosmetic products, by major active ingredients of vegetable origin;

- creating a cosmetic composition in the form of a nourishing, moisturizing and revitalizing day-night cream which, in addition to the skin cleansing effect, also provides elements

The cosmetic product in the form of a cream made represents an association of bioactive principles of vegetable origin that act beneficially, proven in the concentrations used in the proposed formulas, having the action of hydration, revitalization, emollient, stopping the drying and aging of the skin and rebuilding the hydrolipidic film, rendering skin freshness and radiance.

The technical solution consists in associating the concentrated aqueous re-extract from the dessert brown truffle cake with the following ingredients: virgin negrilica (black cumin) oil, Boswellia serrata powder, vegetable glycerin, lavender essential oil and vitamin E.

The advantages of the invention in relation to the state of the art:

The application of the invention leads to the following advantages:

- is a cosmetic product (cosmetic cream composition with nourishing, moisturizing and soothing effect) intended for all skin types, for permanent skin care both for maintenance, having the ability to restore elasticity, being nutritious and revitalizing, as well as for delaying the appearance of the skin aging phenomenon;

- it is an intensive day-night repair cream, formulated for all skin types, even for the driest or senescent;

- it is moisturizing, nourishing and effective for intensifying the skin's natural repair cycle, due to the presence of the aqueous extract of brown dessert truffles and natural negrilica (black cumin) oil, through its antioxidant, anti-inflammatory and nutritional components.

- thanks to the active components with high bioavailability of natural origin, they act against free radicals.

- stimulates cell repair, providing energy, tone and texture of the epidermis.

- the cream with proteins, natural fats and vitamins with a moisturizing and soothing effect for revitalizing the skin contains concentrated aqueous re-extract of brown dessert truffles and virgin negrilica oil (black cumin) which has the ability to restore the skin, nourish the epidermis, to reduce wrinkles, as well as a good cicatrizing and skin projection action.

Thus, the composition increases the density of the dermis and its firmness, has anti-wrinkle, regenerating and restructuring action, protection against the destructive processes triggered by superoxic free radicals and skin toning and has a very good skin acceptability.

Detailed description of the invention:

regarding the composition of bioactive substances of the mentioned components, the motivation of their choice in order to make this cosmetic product is briefly presented

ACTIVE CONSTITUENTS with the potential of exploitation of plant products/by-products intended for valorization/re-valorization

According to the invention, the concentrated aqueous re-extract from brown dessert truffle cake. It is known that brown truffles are very rich in proteins, lipids, minerals, and the valorization of the cake, resulting from obtaining the collar (RO 127733/28.03.2014), which is also very rich in nutritional elements, represents a potential source, as ingredient in cosmetic products, ensuring a special effect, moisturizing and emollient, but also nutritious (thanks to the minerals and vitamins contained), both in the concentrated re-extract from the truffle cake and in the dehydrated cake powder. On the other hand, the use of cake powder, as an ingredient in formulation-development, implies its utilization both as an active ingredient and as a stabilizer, emollient, nutrient and factor for increasing the viscosity of the product. The use of the concentrated re-extract from the truffle cake, which is very rich in minerals, vitamins, lipids and proteins, was used as the aqueous phase in product development-formulation, but also as a solubilizer.

In general, brown truffles are very rich in nutrients, literature data reporting a high content of carbohydrates, up to 60%, protein 20-27%, lipids 3-7.5% (unsaturated and saturated fatty acids), fibers 7-13% and ascorbic acid 2-5%. Besides these, their value is complemented by the content of bioactive compounds, respectively polyphenolic compounds (phenolic acids, phenolic diterpenes), numerous essential minerals have been identified (silicon, potassium, sodium, calcium, magnesium, iron, aluminum, copper, zinc , selenium, chromium) (Enshasy et al., 2013), sugars (mannitol trehalose, raffinose and fructose), but also many essential and non-essential animocides (Kivrak, 2014).

Negrilica (black cumin) virgin oil, obtained by cold pressing black cumin seeds, represents a rich source of nutrients, as a dietary supplement, with a broad spectrum of action, having a composition rich in fatty acids, alkaloids, amino acids, triterpenes , flavones, saponins, steroids, resins, tannins, volatile oils (nigelone and quinone), calcium, potassium, iron, magnesium, selenium, copper, phosphorus, zinc and vitamins.

The vegetable by-product resulting from the cold pressing of black cumin seeds (obtaining virgin oil of negrilica) is very rich in lipids and proteins, which gives it an emollient and moisturizing effect, being exploited, mixtures of black cumin oil , as a constituent part in product formulation-development, respectively in the oily phases. It also brings a considerable contribution, thanks to the minerals and vitamins contained, to the overall intense nutritional effect at the level of the dermis. On the other hand, it is rich in compounds with antimicrobial, antifungal, anti-inflammatory effects, which will make a significant contribution to the quality and value of the products obtained in the framework of the research.

Considering the nutrient-rich contents of the plant products/by-products proposed above, it was intended to include them in the formulation-development of cosmetic/dermetocosmetic products for human use, so that their value is exploited to the maximum for the benefit of the user and to ensure the effects expected therapeutics.

Considering the traditional directions, the plants of the Burseraceae family (eg Boswellia serrata or carterri) have great importance from a therapeutic point of view, they have been used since ancient times. The main components of Boswellia are resins. The most important components from a therapeutic point of view are represented by beta-boswellic acid and its derivatives, being used in the treatment of rheumatism, skin conditions, digestive conditions, for diuretic or neurovegetative effects. Studies have shown definite anti-inflammatory, antioxidant, anti-rheumatic, anti-asthmatic, anti-atherosclerotic, anti-cancer, anti-diarrheal, hepatoprotective, antimicrobial, antifungal, anti-hyperglycemic, restorative, analgesic effects.

Boswellia powder with anti-inflammatory, antibacterial, antifungal effect; obtained in a state of fine powder, by mortaring and sieving. It will be included in the agreed solvent from phase B, respectively in glycerin, even if from the point of view of classification, active principles are included in phase C ingredients. This option was chosen for the best possible incorporation and homogenization of the integral powder of frankincense oleoresin, in order to maximize the exploitation in the formulation of all active biological compounds in the composition of frankincense powder.

The active principle responsible for the pharmaceutical properties of Boswelia serrata extract is beta-boswellic acid.

Vegetable glycerin 99.5% (as a humectant), is a moisturizing agent found in countless cosmetic products for the care and hygiene of the skin and hair. Facilitates the retention of water in the skin tissue, is emollient and velvety. Glycerin can be included in many types of skin and hair care products, in creams, lotions, soaps, toothpaste, shampoos and shower gels. It is easy to work with this ingredient, not being sensitive to heating. Glycerin is not foreign to the human body and thus does not cause irritation or allergies. Glycerin is a good vegetable solvent for various substances, being used in gemotherapy, phytotherapy and homeopathy to extract the active principles. Humectants, like glycerin, are hygroscopic ingredients that draw water from the atmosphere into the skin. In particular, glycerin is skin-friendly, which means that after washing it does not leave the surface of the skin, thus being very useful in hygiene products.

Lavender essential oil is highly valued in aromatherapy and cosmetics; used as a cell regenerator and skin repairer, cicatrizing (helps to restore injured skin, affected by eczema, urticariLxj'șlgi^^ skin soothing (soothes injured, reddened, sunburned or insect-stung skin), antiseptic, antispasmodic, muscle relaxant, antalgic, sedative and soothing, harmonizing, balancing of the nervous system.

Vitamin E is a powerful antioxidant, it maintains a normal metabolism, thus protecting the cells of the body and increasing the production of collagen and elastin, the fibers that support the structure of the skin. Vitamin E helps the skin retain water. acting as a moisturizing product, but also an emollient, leaving a hydrated, velvety and elastic skin; it is the anti-aging vitamin par excellence.

In this sense, the product was created with topical application as a cosmetic cream composition with a nourishing, moisturizing and soothing effect, where the main purpose was represented by the valorization of plant products/by-products generated by some biotechnological processes, by including them as active ingredients or components of emulsion phase. When making these products, the following plant by-products were utilized and included in the product formulas: (i) brown truffle cake, resulting from the cold aqueous extraction of brown truffles, with a concentrated aqueous re-extract included as the working form and fine powder of dehydrated product, (ii) negrilica oil (black cumin), resulting from cold pressing of the seeds, in order to obtain virgin oil, (iii) Boswellia frankincense, as oleoresin in the form of a slightly coarse powder. These were selected and included in the development and formulation of new products, due to the compositions rich in nutritious, emollient, nutritious elements, with biological action known as anti-inflammatory, antibacterial, comforting, antioxidant.

Thus, it can be emphasized that the re-extract from the brown truffle cake was included as an ingredient due to its composition very rich in compounds such as carbohydrates (60%), proteins (27%), lipids such as unsaturated and saturated fatty acids (3 -7.5%), fiber (7-13%), ascorbic acid (2-5%), polyphenolic compounds, minerals such as silicon, potassium, sodium, calcium, magnesium, iron, aluminum, copper, zinc, selenium, chromium and amino acids .

Negrilica oil (black cumin), which was used as the product base for the cosmetic cream formulation, was selected due to its very rich composition in fatty acids, alkaloids, amino acids, triterpenes. flavones, saponins. steroids, resins, tannins, volatile oils, calcium, potassium, iron, magnesium, selenium, copper, phosphorus, zinc and vitamins.

Frankincense, as an anti-inflammatory agent, was selected due to its composition rich in oils, terpenoids, sugars, volatile oils (over 16%), with a high content of betaboswelic acid, alpha-thujene and p-cymene, with anti-inflammatory action.

These were selected and included in the development and formulation of new products, due to the compositions rich in nutritious, emollient, nutritious elements, with known biological action

The invention is presented in more detail by the following example:

Example. In order to obtain 100 g of nutritious, moisturizing and soothing cosmetic cream composition based on organic resins and re-extracts from vegetable by-products (brown truffle cake), use: 20 g of negrilica oil (black cumin), 5 g of emulsifier oil in water (U/A) derived from olive oil, 2 g fatty alcohol co-emulsifier of vegetable origin usually obtained from coconut or palm oil, 68.58 g Re-extract aqueous truffle cake/pure water, 3 g vegetable glycerin, O.lg Boswellia seratta powder, 0.6 g non-paraben, non-formaldehyde, non-isothiazolone eco-certified preservative, 0.45 g lavender essential oil, 0.15 g arnica extract, 0.12 g Vitamin E.

Obtaining the powders of vegetable by-products (truffle cake and incense):

The powder from the truffle cake - the truffle cake resulting from obtaining the collar was stored by freezing until use. After total thawing, the plant material was dehydrated in an oven, for 48-70 hours, at a constant temperature of 60 degrees C. After complete dehydration and collection, the material was crushed by grinding, sieved to remove coarse debris, in order to obtaining a fine and pleasant powder.

Incense powder - the boswellia resin was mortared and then sieved, through a stainless steel sieve, in order to obtain a fine powder with a uniform appearance.

Obtaining the concentrated re-extract from brown truffle cake

The vegetable material (brown truffle cake) stored by freezing, was thawed and subjected to a re-extraction in aqueous medium, by boiling, for 48 hours, after which it was concentrated to half of the final volume obtained. The extract was stored cold, until use in product formulation.

Composition of nourishing, moisturizing and soothing cosmetic cream: qualitative and quantitative formula - INCI-CAS - for 100 g composition (Phase A — lipophilic phase, Phase B — hydrophilic phase, Phase C - active substance)

PHASE NAME AND INGREDIENT Quantity (g) lOOg office house A NigeUu sativa seed oii 20 emollient, moisturizing, antiallergic. antifungal, anti-inflammatory, cicatrizing, skin restructuring 90064-32-7 A Cetearyl olivate, sorbitan olivate 5 emulsifier 223706-40-9 A Cetyl alcohol 2 stabilize 36653-82-4 B Aqua/Truffles Aqueous extract 68.58 solvent 7732-18-5/- B lycerin 3 wet 56-81-5 B Bosellia serrata powder 0.1 inflammatory 97952-72-2 c Benzyl alcohol, salicylic acid, glycerin, sorbic acid 0.6 ecopreservative with antibacterial and antifungal action 100-51-6/69-72- 7/56-81-5/110- 44-1 c Lavandula angustifolia oii 0.45 Soothing, cellular regenerator and skin repair, cicatrizing, antiseptic 90063-37-9 c Helianthus annus seed oii and Arnica montana flower extract 0.15 antibacterial, antirheumatic, anti-inflammatory, calming, antioxidant, anti-irritant, restorative, antalgic, stimulator of peripheral blood circulation 8001-21- 6/68990-11-4 c Tocopherol, Helianthus annuus seed oii 0.12 natural antioxidant, anti-inflammatory, regenerator 8001-21-6

The process of preparing the cream composition according to the invention is carried out in the following way; phase A and B are weighed on the analytical balance, combined, in separate heat-resistant glasses, heated to a temperature of 70°C, in a water bath, until complete melting; glycerin of vegetable origin and incense powder phase B is added over phase A, with homogenization for 3 minutes, with the hand mixer, until complete cooling, phase C is added at the end, after cooling the formed emulsion, then homogenize for another 3 minutes .

After adding phase B over phase A, with homogenization for at least 3 minutes, an emulsion was obtained with a homogeneous appearance, white in color, with a fine, airy texture, with a value of

RELEVANT TESTS FOR SPECIFIC BIOLOGICAL ACTIVITY AND DIRECTION OF DERMATO-COSMETIC APPLICABILITY

ASSESSMENT OF SKIN TOXICITY AND IRRITABILITY

COMPOSITION OF COSMETIC CREAM WITH NUTRITIVE, MOISTURIZING AND CALMING EFFECT

In vitro experimental models were approached for the preclinical pharmacological evaluation of the cosmetic cream composition product with nourishing, moisturizing and soothing effect, in order to establish the basic aspects regarding the product's safety and effectiveness.

The composition of the cosmetic cream with nourishing, moisturizing and soothing effect was tested in accordance with the regulations in force using the Epi-Derm - Skin Irritation Test (EPI-200-SIT) test system, represented by reconstituted human dermis. This model is an alternative test method, validated by ECVAM (European Center for the Validation of Alternative Methods) and one of the test models recommended for the assessment of dermal irritation and corrosivity - Skin Irritation (OECD TG 439) and Corrosion (OECD TG 431) Testing.

Test methodology and results obtained:

Materials and method

The in vitro testing was carried out in accordance with the protocol provided by the company supplying the kit, the company Mattek from Bratislava, distributor of Rites produced by the American company Mattek. (Protocol: In Vitro EpiDerm™ Skin Irritation Test (EPI-200-SIT)).

Test kit (human reconstituted dermis), delivered as viable cultures. 24 pieces of reconstituted human dermis inserts, in a 24-well plate, inserted into the agarose gel and supplemented with culture medium.

Culture medium - DMEM F 12 with 10% fetal serum.

Positive control: sodium dodecyl sulfate (SDS), 0.1 G; it is recommended to dissolve it in saline phosphate buffer, in order to administer, achieving a working concentration of 1%.

Culture plates with 6 wells, culture plates with 24 wells and 96 wells, for carrying out the various experimental stages

According to the EU and GHS classification (category R38/Category 2 or country label), an irritant is predicted if the average tissue viability of three individual tissues exposed to the test substance is

In vitro result Prediction in vivo Average tissue viability <=50% Irritant (I), (R38 or GHS category 2) Average tissue viability >50% Non-irritating (NI)

Results obtained when testing the irritant-corrosive action on a reconstituted dermis model

The viability tests were carried out by applying on the EPI-DERM system, the product Cosmetic cream composition with nourishing, moisturizing and soothing effect, in three series of independent replicates, of 3 sets of determinations in each independent replicate.

The data recorded after performing the test were:

The product Average DO St. dev. p (3) p (4) Relative Viability (%) Conclusion PBS, ¹ 1,970 0.0269 - - 100 SDS, ² 0.024 0.0020 - - 1 Cosmetic cream composition with nourishing, moisturizing and soothing effect 1,514 0.0721 0.0026 3,240 77 Non-irritating (NI)

I: positive control; 2: negative control; 3: student t-test, 2-tailed, homoscedastic distribution. vs. negative control, 4: 2-tailed student t-test, homoscedastic distribution, vs. positive control

In accordance with the classification criteria, it was concluded that the product Cosmetic cream composition with nourishing, moisturizing and soothing effect is not cytotoxic or irritating (NI) and, in conclusion, can be used safely. in accordance with the instructions for use of the products.

Conclusions:

1. The testing carried out qualifies the composition of the cosmetic cream with nourishing, moisturizing and soothing effect for external use, on skin without lesions.

2. The composition of the cosmetic cream with a nourishing, moisturizing and soothing effect is contraindicated for application on open wounds, and it is also not recommended to apply the product by other routes of administration, nor to administer them ocularly or periocularly.

Bibliography. https://www.mattek.com/wp-content/uploads/EPI-200-SIT-Skin-Irritation MK-24-007002310 02 19.pdf

2. OECD Test No. 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method

3. OECD Test No. 431: In vitro skin corrosion: reconstructed human epidermis (RHE) test method

- DETERMINATION OF MICROBIAL LOAD - • Field of applicability: The method is applied in the quality control process of the products to be analyzed: cosmetic cream composition with nourishing, moisturizing and soothing effect and virgin negrilica oil (black cumin).

• Principle of the method: in order to evaluate the microbial load that the products may have, the test method was used with the total number of viable aerobic microorganisms under the conditions provided by the European Pharmacopoeia 9 (FE9). you I, chap. 2.6.12.

• Equipment and glassware: Petri plates with a diameter of 90 mm - sterile, single-use; sterile graduated pipettes of volume 10 ml; 18/180 mm test tubes - sterile; sterile Erlenmayer beakers; automatic pipettor; analytical balance; microbiological cabinet with laminar air flow (LAF); thermostats;

• Culture media used (according to FE in force):

* Casein soya-bean digest agar (CaSoA) - marketed in dehydrated form from Merck Romania;

* Sabouraud with chloramphenicol (SDA) - marketed in dehydrated form from Merck Romania;

^x buffer solution peptone water with sodium chloride. pH 7.

Test methodology with total number of viable aerobic microorganisms - deep inoculation method

The samples are processed in aseptic conditions, in the work point of class A. To test the total number of viable aerobic microorganisms, 10 ml of sample is taken; prepare the 1/10 dilution as follows: add 10 g of the product or 10 ml of the product to 90 ml of peptone water buffer solution with sodium chloride, pH=7 and homogenize; subsequently, serial dilutions (1/100, 1/1000) are prepared, using as a solvent buffer solution peptone water with sodium chloride, pH 7. Each 1 ml of the sample dilution is distributed in 2 sterile Petri dishes with a diameter of 90 mm and add to each plate 15-20 ml of CaSoA culture medium, melted and cooled to a temperature of 43°C.

Cover each Petri dish with the lid, gently rotate it on a flat surface (in order to homogenize the product with the culture medium) and let it solidify at room temperature.

The plates (placed with the lid down) are thermostated at a temperature of 30-35°C for 5

Results x After the incubation period has expired, select the Petri dishes showing microbial growth for each dilution tested;

^x Microbial colonies are counted only on the plates that show microbial growth of a maximum of 250 for the total number of viable aerobic microorganisms (colony-forming units - cfu);

x Microbial colonies are counted only on the plates showing microbial growths of maximum 50 for total number of yeasts and molds;

x Petri dishes with microbial growth are eliminated from the interpretation of results if both plates, corresponding to a tested dilution, indicate the absence of microbial growth;

x The total number of viable aerobic microorganisms is equal to the arithmetic mean of the number of cfu recovered from the CaSoA culture medium; if fungal colonies are also present on this culture medium, they are counted as part of the total number of viable aerobic microorganisms;

The total number of yeasts and molds is equal to the arithmetic mean of the number of cfu recovered from the SDA culture medium, if bacterial colonies are also present on this culture medium, they are counted as part of the total number of yeasts and molds.

Sample result Cosmetic cream composition with nourishing, moisturizing and soothing effect Total number of viable aerobic bacteria, CFU/g <10 CFU/g Total number of yeasts and molds, UFC/g < 10 CFU/g Virgin Negrilica Oil (Black Cumin) Total number of viable aerobic bacteria, CFU/g <10 CFU/g Total number of yeasts and molds, UFC/g < 10 CFU/g

The composition of the analyzed cosmetic cream with nourishing, moisturizing and soothing effect is compliant from the point of view of the microbial load, presenting values below the maximum imposed by FE 9.

The analyzed sample of virgin negrilica oil from black cumin seeds is compliant from the point of view of the microbial load, it presents values below the maximum ones imposed by FE 9.

Conclusions and proposals

Considering the growing interest in preparations of natural origin, the created cosmetic cream composition offers a moisturizing, emollient, nutritious, soothing and anti-inflammatory effect for revitalizing the skin, being effective, stable, pleasant, relatively cheap, easy to administer, apply and use in conditions of safety.

The favorable therapeutic properties result from adding the therapeutic effects of truffle cake extract and oil extracts from a variety of plants to those of a lipophilic cream base consisting of negrilica seed vegetable oil, lavender essential oil and total amica extract , all presenting a certain value and therapeutic importance, which were confirmed by a multitude of data from the specialized literature.

The work carried out on the cosmetic cream composition product with a nourishing, moisturizing and soothing effect demonstrated the following:

- the composition does not show dermal toxicity, as demonstrated by using the in vitro test model EpiDerm™ Skin Irritation Test (EPI-200-SIT)·,

- the presence of pathogenic germs was not revealed, and the microbial load was within the limits imposed by the European Pharmacopoeia for topical preparations;

- obvious anti-inflammatory action, manifested early and maintained during the study (24 hours), on the model of acute edema of the rat's paw provoked with 1% carrageenan solution, for the composition of the cosmetic cream with nourishing, moisturizing and soothing effect, administered repeatedly.

The cream according to the invention, based on the concentrated aqueous re-extract from brown dessert truffle cake, negrilica oil (black cumin), through the active components with high bioavailability of natural (vegetable) origin, following the researches it was found that, presents an obvious anti-inflammatory action, with a moisturizing effect, being at the same time emollient and nourishing, for revitalizing the skin, truly unique; it preserves the elasticity and integrity of the skin, also having an anti-wrinkle action, which prevents the formation of small skin wrinkles and restores the epidermal integrity (being an intensive day-night repair cream, for all skin types), with the hydration of the areas on which it is applied, with effect synergy.

8araka

Claims (1)

COMPOSITION OF COSMETIC CREAM WITH NUTRITIONAL, MOISTURIZING AND CALMING EFFECT CLAIM Cosmetic cream composition with a nourishing, moisturizing and soothing effect, characterized by the fact that it consists of: 20 g of negrilica oil (black cumin), 5 g of oil-in-water emulsifier (U/A) derived from olive oil, 2 g of co -fatty alcohol emulsifier of plant origin usually obtained from coconut or palm oil, 68.58 g Re-extract aqueous truffle cake/pure water, 3 g vegetable glycerin, O.lg Boswellia seratta powder, 0.6 g non-paraben, non-formaldehyde, non-isothiazolone eco-certified preservative, 0.45 g Lavender essential oil, 0.15 g Arnica extract, 0.12 g Vitamin E.