Pigment comprising gardenia blue

Abstract

A blue pigment for cosmetic applications is provided, which exhibits excellent color and stability characteristics, while being fully plant-based, edible and of natural origin.

Description

[0001] PIGMENT COMPRISING GARDENIA BLUE

[0002] The present invention relates to a blue pigment for cosmetic applications, in particular for make-up compositions.

[0003] Cosmetics are products used to enhance or change the appearance of the face, fragrance or the texture of the body. They are generally mixtures of chemical compounds derived from natural sources (such as coconut oil), or may be synthetic or artificial. Cosmetics that are applied to the face to enhance one's appearance are also known as make-up, which include items such as lipstick, mascara, eye shadow, foundation, blush, highlighter, bronzer and several other products.

[0004] In the United States, the Food and Drug Administration (FDA), which regulates cosmetics, defines cosmetics as "intended to be applied to the human body for cleansing, beautifying, promoting attractiveness, or altering the appearance without affecting the body's structure or functions". This broad definition includes any material intended for use as an ingredient of a cosmetic product.

[0005] Color cosmetics or make-up encompasses different categories of products for skin, eyes, cheeks and lips. It is a growing market as consumers increasingly seek to be always selfie-ready. Color cosmetics can enhance overall physical appearance, conceal flaws or define features, making one appear healthier and look more refreshed. Consumers have also expressed that color cosmetics have helped improve their mental well-being, by empowering them and boosting their selfconfidence.

[0006] The four main categories are: face make-up, such as foundation, blushers, illuminators, face bronzing lotions, creams and powders, loose and pressed powders, and mineral powders; eye make-up, such as eye shadows, eyeliners, eyebrow pencils, kohl and mascara, and mineral powders; lip make-up, such as lipstick, lip glosses, lip pencils, lip plumpers, pots and palettes; and nail make-up, such as nail varnishes and polishes, hardeners and strengtheners, base and top coat.

[0007] Lip color is currently the most active color cosmetics category, accounting for 32% of new launches with over 900 million lipsticks sold worldwide every year. Key performance attributes sought by consumers are color (93%), duration or long-lastingness (90%), texture (90%) and finish (89%).

[0008] Today, the blue pigment used in most color cosmetics on the market are either synthetic or mineral.

[0009] There is, however, a growing consumer demand for cleaner, healthier, more natural and more sustainable beauty products. 32% of consumers state that natural / organic make-up is their first choice, and 63% like it but not always buy it. In particular young consumers are looking for vegan alternatives.

[0010] But consumers are not willing to compromise on performance, even for a 100% natural lipstick. Less than 15% would accept reduced performance with regard to color, coverage and finish. In particular, long-lastingness is clearly an unmet consumer need.

[0011] Blue color is not very common in nature, and the main available botanical or algae extracts are water- soluble, meaning that they are not compatible with anhydrous-oily cosmetic formulations.

[0012] It is therefore an objective of the present invention to provide plant-based pigments of natural-origin, which exhibits excellent color, coverage, long-lastingness and finish characteristics.

[0013] This problem has been solved by the pigment and cosmetic composition of the present invention.

[0014] In a first aspect, the present invention provides a pigment for use in cosmetic applications, comprising micronized Gardenia Blue and kaolinite clay.

[0015] The pigment of the invention is of 100% natural origin according ISO16128, cosmos approved, and China compliant. More importantly, it is easily dispersible in anhydrous cosmetic formulae, has a high intensity (chroma C*) and coverage (Cr*), as well as a good spreadability, and is stable to light and temperature.

[0016] Surprisingly, the pigment of the invention was found to perform even better than commercially available, non-natural alternatives in certain applications.

[0017] In a second aspect, the present invention provides a cosmetic composition comprising the pigment of the invention and a cosmetically acceptable excipient.

[0018] Gardenia Blue is a natural pigment that is obtained from the Gardenia fruit. It is a dark blue powder that can easily dissolve in water, ethanol solution and propylene glycol solution. This product is often used food products.

[0019] Throughout this disclosure, the terms “Gardenia Blue” and “Gardenia Blue Extract” are used interchangeably, unless otherwise noted.

[0020] Gardenia Blue is typically obtained from Gardenia fruit extracts, which are typically of a brownish color, by hydrolysis and subsequent reaction with an amine. Gardenia fruits comprise an iridoid glycoside called geniposide, the hydrolysis of which affords the respective aglycone genipin. Genipin (IUPAC name: methyl (1 R,4aS,7aS)-1-hydroxy-7-(hydroxymethyl)-1 ,4a,5,7a-tetrahydro- cyclopenta[c]pyran-4-carboxylate) has the following chemical structure:

[0021] Genipin is believed to react with amines to form a blue complex molecule - Gardenia Blue. For example, such a complex has been described in J. Agric. Food Chem. 2021, 69, 3904-3911 , “Molecular structure of Gardenia blue pigments by reaction of Genipin with benzylamine and Amino acids”.

[0022] Without being bound by theory, it is believed that the reaction of genipin with the amine involves dehydration, polymerization and oxidation, to afford a complex molecule of the following structure:

[0023] During the extensive studies leading to the present invention, several natural dyes derived from botanical extracts (persicaria tinctoria (indigo), daucus carota L , ipomoea batatas L. (purple sweet potato), brassica oleracea L. var rubra (red cabbage), Genipa americana L.) were tested, and Gardenia Blue was found to provide the best results. Surprisingly, it was found that the addition of kaolinite clay allowed to increase the C* (intensity) of the pigment.

[0024] Furthermore, micronization was found to improve its spreadability.

[0025] Kaolinite clay (also called kaolin) is a clay mineral with the chemical composition AI2Si2O5(OH)4. It is a layered silicate mineral, with one tetrahedral sheet of silica (SiO4) linked through oxygen atoms to one octahedral sheet of alumina (AIO6). Kaolinite is a soft, earthy, usually white, mineral, produced by the chemical weathering of aluminium silicate minerals like feldspar.

[0026] Throughout this disclosure, the terms “kaolinite”, “kaolinite clay”, and “kaolin” are used interchangeably unless otherwise noted. It may also be identified by the CAS number 1332-58-7.

[0027] During the extensive studies leading to the present invention, several clays were evaluated. Surprisingly, it was found that kaolinite provided the best stability and highest intensity. This finding is contra-intuitive, as kaolinite has an inactive nature based on its structure. Furthermore, it was described in the prior art (e.g. Shue Li, Bin Mu, Xiaowen Wang, Aiqin Wang, “Recent researches on natural pigments stabilized by clay minerals: A review”, Dyes and Pigments, Volume 190, 2021 , 109322, ISSN 0143-7208) that kaolinite is not very suitable for stabilizing natural pigments.

[0028] As an alternative or in addition to kaolinite, montmorillonite may also be used in the pigment of the invention.

[0029] It has been found that by micronizing the Gardenia Blue, and optionally also the kaolinite clay, a highly uniform pigment with good dispersibility and spreadability can be obtained.

[0030] In an embodiment, the Gardenia Blue is obtained from a Gardenia florida L. fruit extract.

[0031] Gardenia florida L. Rubiaceae is commonly called Cape jasmine or Gardenia. There are also several alternative names, including Gardenia jasminoides J. Ellis, Gardenia augusta Merr, Gardenia florida L., Gardenia radicans Thunb., Varneria augusta L., and Fructus gardenia.

[0032] Throughout this disclosure, the terms “Gardenia”, “Gardenia florida L ", “Gardenia florida L. Rubiaceae”, “Gardenia jasminoides J. Ellis”, “Gardenia augusta Merr”, “Gardenia radicans Thunb ", “Varneria augusta L", “Fructus gardenia”, and “Cape jasmine” are used interchangeably unless otherwise noted.

[0033] Also, unless otherwise note, any Gardenia extract refers to the Gardenia fruit extract.

[0034] Gardenia florida L is a small shrub that can grow up to twelve meters tall, but is usually much shorter, with greyish bard, evergreen leaves. The fragrant flowers are white and have a matte texture in contrast to the glossy leaves. The fruits have an orange-red color. Gardenia florida L is growing wild and also cultivated in warm temperate and subtropical climates. Typical countries of origin include Asian countries, such as e.g. Vietnam, China, Taiwan, Japan, Myanmar and India.

[0035] Gardenia extracts have been used as natural yellow or blue dye for many years, but they also have various biological activities, such as antidepression, antioxidant properties, and improvement of the quality of sleep after oral administration.

[0036] Gardenia florida L. fruit extract typically comprises a number of components, which may slightly vary depending on origin, season and production method.

[0037] Among the extract’s components, geniposide and any other genipin derivatives are highly relevant for the blue color.

[0038] Other typical components include, but are not limited to, jasminosides and jasminoside derivatives, syringinoside, crocusatin C, blumenol A, gardenone and gardenone derivatives, sugars and oligosaccharides.

[0039] Hydrolysis of the Gardenia florida L. fruit extract and subsequent treatment with an amine is preferably used to obtain the Gardenia Blue.

[0040] Therefore, in an embodiment, the Gardenia Blue is obtained from a Gardenia florida L. fruit extract by hydrolysis and treatment with an amine, in particular with a primary amine.

[0041] Hydrolysis of geniposide or any other genipin glycosides present in Gardenia florida L. fruit extract can be affected by known methods. Enzymatic hydrolysis, in particular with a glucosidase, was found to be particularly effective.

[0042] Therefore, in an embodiment, the hydrolysis is an enzymatic hydrolysis, and in particular an enzymatic hydrolysis including a glucosidase.

[0043] Suitable enzymes are generally known and commercially available. For example, Pectinex XXL (from Novozymes) may be used.

[0044] In an embodiment, the amine is selected from an amino acid, an amino acid salt, a peptide, a protein, and mixtures thereof. It has been found that these amines provide a pigment with a particularly interesting blue color. Preferably, the amine is selected from glutamine, a glutamate salt, and mixtures thereof. Glutamine and glutamine derivatives were found to provide a very strong blue color. In comparison, glycine and glycine derivatives provided a blue-purple shade and lysine, arginine and their derivatives provided a greener shade.

[0045] For example, monosodium glutamate may be used. Monosodium glutamate is commercially available, and may be derived from e.g. corn by fermentation.

[0046] In order to obtain a strong blue color, it is advantageous if the Gardenia florida L. fruit extract comprises a relatively high amount of geniposide and / or other genipin glycosides.

[0047] Therefore, in an embodiment, the Gardenia florida L. fruit extract comprises from about 2 wt% to about 12 wt% of geniposide, more preferably from about 4 wt% to about 10 wt% of geniposide, and most preferably from about 6 wt% to about 8 wt% of geniposide, based on the dry weight of the Gardenia florida L. fruit extract.

[0048] After hydrolysis and reaction with the amine, i.e. in Gardenia Blue, there should be no or almost no genipin left in the material, as genipin could potentially react with amino acids and other amines on human skin.

[0049] Therefore, in an embodiment, the Gardenia Blue comprised in the pigment of the invention is free or at least essentially free of genipin.

[0050] In other words, in an embodiment, the Gardenia Blue comprises less than about 12 ppm of genipin, more preferably less than about 10 ppm of genipin, still more preferably less than about 5 ppm of genipin. Most preferably, the Gardenia Blue does not comprise any genipin, or at least no genipin is detected in the Gardenia Blue.

[0051] Analysis of the Gardenia Blue, and in particular the determination of its genipin content, may be done by any suitable method, e.g. by HPLC-UV, HPLC-MS, HPLC-DAD, HPLC-ELSD, or HPLC- Amperometry. HPLC-DAD for example, which uses a Diode Array Detector, allows for measuring in the UV-visible area. Genipin is typically detected at a wavelength of about 242 nm. Therefore, preferably, no genipin is detected in the Gardenia Blue when using either one or several of these methods.

[0052] Suitable methods for obtaining Gardenia florida L fruit extracts are generally known in the art (e.g.

[0053] Phytochemistry review 2016, 15:37-49. A review through recovery, purification and identification of genipin; or chapter 20 “Natural Solutions for blue colors in food, Handbook on Natural Pigments in Food and Beverages 2016 355-384).

[0054] For example, Gardenia florida L fruits (fresh or dried) may be crushed or otherwise reduced to smaller pieces and then extract with water, preferably with hot water (e.g. between about 50 °C and about 75 °C). The crude extract may be filtered. Optionally, it is purified by adding ethanol or an ethanol-water mixture and a resin (e.g. a macroporous ion exchange resin) to obtain a purified extract. The pH of the extract may be adjusted, for example by the addition of an acid (e.g. citric acid) or a base (e.g. sodium carbonate).

[0055] A liquid extract may be subjected to concentration, spray drying, and / or other drying techniques known in the art to obtain a solid extract.

[0056] Preferably, a solid extract is used for the preparation of the pigment of the invention.

[0057] Gardenia florida L. fruit extracts are also commercially available, e.g. Gardenia Blue Color from CNJ Nature Co. Ltd. in China.

[0058] The Gardenia Blue may be purified prior to use, e.g. by heat treatment, treatment with ethanol, treatment with a resin, UV irradiation, filtration, or a combination of any such treatments.

[0059] The Gardenia Blue used in the pigment of the invention is micronized. It has been found that micronization has a positive impact on chroma and coverage.

[0060] The most commonly used metrics when describing particle size distributions are D-Values: D10, D50 and D90, which are the intercepts for 10%, 50% and 90%, respectively, of the cumulative distribution. The D50 is the diameter of the particle that 50% of a sample's mass is smaller than and 50% of a sample's mass is larger than. In other words: D represents the diameter of powder particles, and D50 means a cumulative 50% point of diameter (or 50% pass particle size); D10 means a cumulative 10% point of diameter. D50 is also called average particle size, median diameter, mass-median-diameter (MMD) or log-normal distribution mass median diameter.

[0061] D50 is also divided into Dv50, Dw50 and Dn50; Dv50 means volume D50, whereas Dw50 is mass D50 and Dn50 is number D50. Dv50 is also known as volume median or volume average particle size, it physically represents that each volume of particles greater or smaller than such value takes account of 50% of the total particles volume. Dn50 is known as number median, it physically represents that each number of particles greater or smaller than such value takes account of 50% of the total particles number. In an embodiment, the micronized Gardenia Blue has a volume median particle size (Dv50) of from about 1.0 pm to about 7.5 pm, more preferably of from about 1.5 pm to about 6.0 pm, and most preferably from about 2.0 pm to about 5.0 pm.

[0062] In an embodiment, the micronized Gardenia Blue has a volume particle size 90% percentile (D90) of from about 3.0 pm to about 20.0 pm, more preferably of from about 4.0 pm to about 17.0 pm, and most preferably from about 5.0 pm to about 15.0 pm.

[0063] Depending on the intended use or application, the ratio between the micronized Gardenia Blue and the kaolinite clay may be selected by the skilled person. For most cosmetic applications, a weight ratio in which the two components are used in amounts of similar magnitude is suitable.

[0064] In an embodiment, the weight ratio of micronized Gardenia Blue to kaolinite clay is from about 1 :4 to about 1 :1 , more preferably from about 1 :3 to about 1 :2, and most preferably about 1 :2.5. These ratios have been found to be particularly suitable for color cosmetic applications, such as lipsticks, eye shadows or compacted powders, for example.

[0065] The pigment of the invention typically has a blue color, with varying shade depending on the concentrations of the components used.

[0066] For example, the pigment may provide a blue color similar to Pantone 2168 U (CMYK: 26; 11 ; 0; 54 / / sRGB: 87; 104; 117 / / Hex: #576875 / / Light Reflectance Value (LRV) = approx. 13).

[0067] The shade will also somewhat vary depending on the amount of pigment used in a cosmetic formulation.

[0068] In order to adjust the shade to the desired color, it is also possible to include a color-adjusting substance in the pigment of the invention, for example titanium dioxide (TiO2) or zinc oxide for obtaining a lighter shade or CI77007 Ultramarines, Blue 1 Lake or Carbon Black (Cl 77268:1 ; charcoal) to obtain a darker shade.

[0069] It may also be mixed with other pigments to obtain mixed colors, for example purple by mixing with a red pigment or green by mixing with a yellow pigment. Preferably, such other pigments are also of natural origin.

[0070] The pigment of the present invention may further comprise a suitable carrier. Therefore, in an embodiment of the present invention, the pigment of the present invention further comprises a polysaccharide (e.g. maltodextrin or hydrolyzed corn starch), gum arabic, starch, cellulose, silica, calcium stearate, calcium carbonate, tricalcium phosphate, glucose syrup, or any other suitable carriers or mixtures thereof.

[0071] In an embodiment, the pigment of the invention comprises a polysaccharide, more preferably maltodextrin. Such a carrier may already be included for a spray drying step, if applicable, and allows for improving the flowability and for avoiding caking and stickiness. Typically, if maltodextrin is included, it is included in slightly higher amounts than the Gardenia Blue and the kaolinite clay.

[0072] For instance, the concentration of micronized Gardenia Blue in the pigment can be from about 5 wt% to about 20 wt%, more preferably from about 10 wt% to about 15 wt%, for example about 12 wt% of micronized Gardenia Blue.

[0073] For instance, the concentration of the kaolinite clay in the pigment can be from about 20% to about 40%, more preferably from about 25% to about 35%, for example about 30% of kaolinite clay.

[0074] For instance, the concentration of the maltodextrin in the pigment can be from about 50% to about 65%, more preferably from about 55% to about 60%, for example about 58% of maltodextrin.

[0075] In an embodiment, the pigment comprises from about 5 wt% to about 20 wt% of micronized Gardenia Blue, from about 20 wt% to about 40 wt% of kaolinite clay, and from about 50 wt% to about 65 wt% of maltodextrin; more preferably comprising from about 10 wt% to about 15 wt% of micronized Gardenia Blue, from about 25 wt% to about 35 wt% of kaolinite clay, and from about 55 wt% to about 60 wt% of maltodextrin, for example about 12 wt% of micronized Gardenia Blue, about 30 wt% of kaolinite clay, and about 58 wt% of maltodextrin.

[0076] The positive effects of micronization have been discussed above.

[0077] In an embodiment of the present invention, not only the Gardenia Blue is micronized, but also the kaolinite clay and, if present, any other solid components. Preferably, most of the solid components are micronized. More preferably, at least all solid components of the pigment are micronized.

[0078] In an embodiment of the present invention, the entire pigment is micronized. The micronization can be done either on the separate components, on several partial mixtures, or on the full composition. It is also possible to perform several micronization steps. In a second aspect, the present invention provides a cosmetic composition comprising the pigment of the invention and a cosmetically acceptable excipient.

[0079] Preferably, the cosmetic composition comprises the pigment of one or more of the specific embodiments outlined above.

[0080] Any excipients commonly used in the preparation of cosmetic preparations for use on the human skin may be employed in the present invention. Suitable excipients include, but are not limited to ingredients that can influence organoleptic properties, dispersibility, appearance, and spreadability of the pigment of the present invention. More specifically, they include liquids, such as water, oils or surfactants, including those of petroleum, animal, plant or synthetic origin, such as and not restricted to, peanut oil, soybean oil, mineral oil, sesame oil, castor oil, polysorbates, sorbitan esters, ether sulfates, sulfates, betaines, glycosides, maltosides, fatty alcohols, nonoxynols, poloxamers, polyoxyethylenes, polyethylene glycols, dextrose, glycerol, digitonin, squalene, hemisqualene, and the like, as well as mixtures thereof.

[0081] The cosmetic composition may take any physical form and may be produced in any solid, liquid, or semi-solid form useful for application.

[0082] In an embodiment, the cosmetically acceptable excipient comprises castor oil. More preferably, the cosmetically acceptable excipient at least essentially consists of castor oil. Castor oil is of natural origin and has been found to provide a good dispersion of the pigment of the invention.

[0083] In an embodiment, the cosmetically acceptable excipient comprises squalene and / or hemisqualene. More preferably, the cosmetically acceptable excipient at least essentially consists of squalene and / or hemisqualene. Squalene and / or hemisqualene have been found to provide a good dispersion of the pigment of the invention.

[0084] In an embodiment, the cosmetically acceptable excipient comprises a mixture of castor oil with squalene and / or hemisqualene.

[0085] The cosmetic composition of the present invention is typically a color cosmetic or make-up composition. Possible product formats have been mentioned in the introductory section above.

[0086] In an embodiment, the cosmetic composition of the present invention is selected from the group consisting of a lipstick, a lip balm, a lip oil, a lip gloss, a lip scrub, a make-up emulsion, a make-up powder, a make-up oily gel, an eye shadow, a compacted powder, a nail polish, a mascara, a foundation, and a soap. In a preferred embodiment, the cosmetic composition of the invention is a lipstick composition, an eye shadow composition or a compacted powder composition.

[0087] The pigment of the invention was found to provide collagen boosting and wound-healing activity.

[0088] In view of this biological activity, the pigment of the invention is also highly beneficial for skin care applications.

[0089] Therefore, in an embodiment, the cosmetic composition of the invention is a skin care composition.

[0090] The present invention is further illustrated by means of the following non-limiting examples:

[0091] 1 : Preparation of Pigment

[0092] Gardenia florida L. fruit extract is commercially available from several suppliers, for example Gardenia Blue Color from CNJ Nature Co. Ltd. in China.

[0093] Kaolinite clay is commercially available from several suppliers, for example from Color Clay Natural S.L. in Spain.

[0094] In order to prepare the pigment of the invention, 280 kg of a Gardenia florida L. fruit extract powder were dispersed in 600 kg of tap water and 120 kg of kaolinite clay were added. The mixture was stirred at 85 °C during about 1 hour until homogeneisation.

[0095] The obtained mixture was spray dried with a spray dry apparatus (Anhydro spx flow) and sieved (mesh size 1000 pm) to obtain 360 kg of powder. Micronization (e.g. by air jet micronization) of the powder afforded 342 kg of micronized powder.

[0096] The thus obtained pigment had a particle size distribution of Dv10 = 1.24 pm, Dv50 = 3.39 pm, and Dv90 = 6.86 pm. Characteristics of Pigment

[0097] Four different batches of pigments according to Example 1 were assessed and found to exhibit the following characteristics:

[0098] The colorimetric properties were measured through L,a,b measurements (Spectrocolorimeter Konica-Minolta 3600d (D65, L*, a*, b*, h, C*, Y)). L,a,b colorimeters are widely used to evaluate color variations of products. Any color shade can be specified by three coordinates in the L*,a*,b* three- dimensional system:

[0099] - the L* axis runs from 0 (black) to 100 (white)

[0100] - the a* axis (green / red direction)

[0101] - the b* axis (blue / yellow direction).

[0102] Positive a* is red, and the higher the a* value, the more intense the red. Analogously, negative a* is green, positive b* is yellow, and negative b* is blue.

[0103] L* is the clarity value, representing darkness to lightness, with values ranging from 0 to 100. The 50% light level is considered "normal" and is often used to display colors.

[0104] Y refers to the relative Luminance. This is a photometric measure of the luminous intensity per unit area of light travelling in a given direction. It is used to determine the opacity or coverage.

[0105] Opacity is a measure of covering power, 100% opacity means full coverage.

[0106] Opacity / coverage (CR) is calculated as follows: CR (%) = ((Y black*100) / Y white)

[0107] Chroma (C*) is the amount of saturation of a color. Colors of high chroma are said to be clear, bright or brilliant. Dull (pastel) colors have a low chroma. Lower saturation means that the color has more grey and appears faded and less intense.

[0108] Hue (h) is the color tone or color name of a color. It is the property of light which identifies color within the spectrum. Hue ranges over colors with familiar names like red, orange, yellow, green, blue, indigo, and violet. For convenience, hue is often visualized to be on a color wheel or circle. In Web colors, hue is measured in 360 degrees of a color circle. Red is at 0°, green is at 120°, and blue is 240°. At 360°, the measure of hue goes back to 0°. Example s: Stability Testing

[0109] Batch No. 1 from Example 2 was submitted to stability testing under various conditions. The results are shown in the following two tables: As can be seen from the above, whatever the conditions, the pigment of the invention is very stable, with a dE2000 that is below 1.0, which confirms that the color change is not visible to the eye.

[0110] Example 4: Effect of Micronization

[0111] In order to assess the effect of micronization, samples of spirulina extract and kaolin (both commercially available) were micronized and compared to the respective non-micronized materials and to the pigment of the present invention (gardenia blue extract / kaolin 70:30, micronized).

[0112] The colorimetric properties were measured through L,a,b measurements (Spectrocolorimeter Konica-Minolta 3600d (D65, L*, a*, b*, h, C*)). The granulometry of the gardenia-containing samples was determined using a particle size analyser Mastersizer 3000+ Ultra from Malvern Panalytical.

[0113] As can be seen from the above table, micronization allowed to modify the colorimetric parameters and to increase the Chroma C*, the L* and the b* (more blue) of the gardenia blue extract, but has hardly any impact on the kaolin. With the mixture with kaolin, micronization also allowed to modify the colorimetric parameters and to increase the C*, the L* and the b (more blue).

[0114] Furthermore, the particle size of the micronized samples 2 and 6 is clearly more uniform than that of the non-micronized samples 1 and 5.

[0115] Photographs of the powder samples 1 , 2, 5, and 6 are shown in Figure 1 . As can be seen from these photographs, the addition of kaolin combined with micronization (sample 6) led to a real difference in terms of the colorimetric parameters compared to the other three samples.

[0116] Example 5: Comparison of Clays

[0117] In order to assess the impact of the clay used, the following two commercially available clays were tested:

[0118] - Montmorillonite: CosWHITE™ (supplied by Argile du Velay)

[0119] - Kaolin: CosKAO™ (supplied by Argile du Velay)

[0120] Gardenia blue extract, kaolin and montmorillonite, respectively, were subjected to micronization to obtain comparative samples.

[0121] For the mixtures, 500 g of gardenia blue were dispersed in 2 kg of tap water, and 500 g of clay was added under stirring. The mixture was then spray dried and the resulting powder micronized to obtain the test samples.

[0122] The colorimetric properties were measured through L,a,b measurements (Spectrocolorimeter Konica-Minolta 3600d (D65, L*, a*, b*, h, C*)).

[0123] As can be seen from the above table, the addition of montmorillonite to the gardenia blue extract led to a decrease in Lightness, and an increase of Chroma C* and b* (more blue).

[0124] As can be seen from the above table, the addition of Kaolin to the gardenia blue extract led to an increase in Lightness, an increase of Chroma C* and b* (more blue). In addition, a stability testing revealed that the best ratio for kaolin is with the lowest tested percentage of kaolin for compositions comprising gardenia blue extract and kaolin:

[0125] At 105 °C and after suntest, the least stable prototype was the one with the highest percentage of kaolin (70%). This observation is really surprising because clay is generally described to stabilize coloring extracts: A review”, Dyes and Pigments, Volume 190, 2021 , 109322, ISSN 0143-7208).

[0126] Example 6: Performance of Lipstick Compositions

[0127] In order to determine the coloring properties and performance in formulation, the pigment of the invention was formulated in a lipstick base at different concentrations of the pigment of Example 1 and with varying amounts of titanium dioxide (Samples 1-5).

[0128] For comparison, Unipure Blue LC621 (Blue 1 Lake; synthetic pigment) and Unipure Blue LC680 (INCI: CI77007 Ultramarines & kaolin; mineral pigment) were also formulated in the same lipstick base (Samples 6 and 7), as well as Natpure Xfine Spirulina SL615 (Sample 8; Spirulina platensis extract & trehalose dihydrate).

[0129] Sample 9 has the same composition as Sample 1 , but was further spread on a contrast card. Samples 10 to 12 were also spread on contrast cards, and comprise the comparative pigments Unipure Blue LC621 (Sample 10), Unipure Blue LC680 (Sample 11) were also formulated in the same lipstick base and Natpure Xfine Spirulina SL615 (Sample 12), respectively, at 10 wt%.

[0130] Phase A was prepared by blending butyrospermum parkii butter, cera alba, caprylic / capric triglycerides, linoleic acid / propanediol copolymer, jojoba esters and helianthus annuus seed wax and acacia decurrens flower wax and polyglycerin-3 at a temperature of 80 °C until homogeneous.

[0131] To obtain phase B, a mixture of the respective pigment powder and the castor oil was homogenized in a turrax blender for 2 min at room temperature.

[0132] The two phases were combined at 75 °C and blended in the turrax blender for 2 min.

[0133] The obtained lipstick composition was cooled down to room temperature for 2 hours under magnetic agitation.

[0134] Finally, for Samples 9-12, 2 g of the thus obtained lipstick compositions were crushed between glass slabs and applied to a contrast card using a film applicator (film thickness: 50 pm).

[0135] L*,a*,b* measurements were performed on a Spectrocolorimeter Konica-Minolta 3600d (D65, L*, a*, b*, h, C*) and DeltaE(2000) was measured to see the color and opacity differences between the different pigments in lipstick base. CR / opacity was only determined for Samples 9-12.

[0136] The results are shown in the following table:

[0137] The comparison of Samples 1 , 2, 3 and 4 demonstrated that the pigment of the invention provides access to a variety of shades of blue, depending on how much TiO2is included. Increasing the amount of TiO2allowed to increase the Lightness L* and to obtain a variety of hues (h).

[0138] A comparison of Sample 3 and Sample 8 demonstrated that the pigment of the invention provides the same hue (h) as a synthetic pigment.

[0139] Example 7: Evaluation of Anti-ageing Activity

[0140] The aim of this study was to evaluate the anti-ageing activity of the pigment of the invention. The anti-ageing activity was evaluated by the guantification of type I pro-collagen release in premature aged condition, which was mimicked by chemical treatment (H2O2). Treatment with H2O2is largely described in the literature to induce cells’ premature ageing through the induction of senescence. In the present study, this condition was used as the “aged” condition.

[0141] Cell Culture and Treatment

[0142] The cell culture was done on primary cells isolated from biopsies. Normal Human Dermal Fibroblasts (NHDFs) were seeded in a 96-wells black plate with glass bottom at 10’000 cells per well in triplicate. The cells were incubated for 48 hours in complete medium (DMEM medium, Gibco) supplemented with 10% fetal calf serum (FCS, Biowest) and 1% antibiotics (Sigma-Aldrich) at 37 °C with 5% CO2.

[0143] At the end of the incubation, the cells intended for the “aged” condition were stressed by treatment with hydrogenperoxide (H2O2, Sigma-Aldrich) at 500 pM for 2 hours at 37 °C with 5% CO2.

[0144] Cells were then rinsed twice with PBS (Gibco) and incubated for 72 hours in basal medium (DMEM medium without FCS) supplemented with 1 % antibiotics with the following conditions: The actives were diluted in culture medium.

[0145] Skin cells untreated and cultivated with basal medium were used as a negative control.

[0146] Cells were then incubated for 72 hours at 37 °C, 5% CO2.

[0147] At the end of the culture, a collagen I immunostaining assay was performed as described below.

[0148] Collagen I Immunostaining

[0149] Cells were fixed for 5 minutes with 2% paraformaldehyde (PAF), rinsed with PBS and permeabilized for 15 minutes with 2% PAF + 0.5% Triton X100. Cells were then rinsed again with PBS and nonspecific sites were saturated for 1 hour with a saturation solution (3% bovine serum albumin + 0.1 % Tween 20). Cells were incubated overnight at 4 °C with a primary anti-collagen I antibody from rabbit (Abeam) diluted at 1 :500 in saturation solution diluted 1 :10.

[0150] On the next day, the primary antibody was removed and the cells were rinsed three times with washing solution (PBS + 0.3% Triton X100) and two times with PBS before being incubated for 2 h 30 min at room temperature with a secondary anti-rabbit antibody Alexa Fluor 488 diluted at 1 :500 and Hoechst 33342 diluted 1 :5000 in saturation solution diluted 1 :10. After secondary antibody incubation, the antibody solution was removed and cells were rinsed two times with PBS.

[0151] Pictures were taken with PICO Cell Reporter Xpress (Molecular Devices) in DAPI and Green channels in each well. Automatized cell scoring was performed, and the % of cells expressing collagen I was compared for each of the conditions.

[0152] Statistical Analysis

[0153] A Shapiro- Wilk normality test was performed to evaluate whether the data follow the Gaussian Law. The results did not follow the Gaussian Law.

[0154] As a consequence, a non-parametric statistical analysis was performed by Kruskal- Wall is ANOVA followed by Mann Whitney U test. Results were considered significant with p<0.05 with *, p<0.01 with ** and p<0.001 with ***.

[0155] Results

[0156] The results are expressed in % of the condition, which received only the H2O2stimulation and was identified as “Aged” condition, and are shown in the following table:

[0157] As can be seen from the above, the chemical stress (H2O2; identified as “aged”) induced a significant decrease in the number of positive cells for collagen I expression, indicating a lower capacity of collagen synthesis by the fibroblasts.

[0158] In this stressed model, the stimulation of type I pro-collagen synthesis was still possible as demonstrated by the increase of cells expression collagen I by +268%** in presence of TGF-p at 10 ng / ml and ascorbic acid at 20 ng / ml.

[0159] In the same conditions, the blue pigment of the invention significantly increased the number of positive cells for collagen I expression, by up to +78%**.

[0160] The above results confirm that the blue pigment of the invention, at 0.1 mg / ml, exhibits an anti-ageing activity in this experimental condition by improving the collagen I expression. It was further observed that the lower concentration of 0.05 mg / ml showed the same activity, meaning that the maximal effect on collagen I synthesis was already reached from 0.05mg / ml.

[0161] Example 8: Evaluation of Wound Healing Effect

[0162] The aim of this study was to evaluate the wound healing effect of the pigment of the invention in a culture of keratinocytes. To this end, a percentage of cell migration surface was assessed after scratching the cell layer and after 8 hours of wound induction.

[0163] Cell Culture and Treatment

[0164] The cell culture was realized with primary cells isolated from fresh biopsies. Normal Human Keratinocytes (NHKs) were seeded in a type I collagen pre-coated 24 wells-plate at 50’000 cells per well in quadruplicate. After 48 hours of incubation in complete medium (Dermalife medium, supplemented with Life factors K, Cell Systems) and 1% of antibiotics (Sigma-Aldrich) at 37 °C with 5% CO2, the cells were rinsed twice with PBS (Gibco) and incubated for another 8 hours in basal medium. At the end of the incubation, the cells were treated overnight with the active ingredients at 37 °C with 5% CO2. The cells were treated in basal medium with the following conditions:

[0165] The test substances were diluted in culture medium.

[0166] Skin cells untreated and cultivated with medium were used as a negative control.

[0167] The next day, 30 minutes prior to the scratch, a solution of calcein AM (Abeam) at 1 pM was added in the supernatant, in order to label the living cells. Then, a scratch was realized manually using a culture cone on the cell layer and then rinsed twice with PBS in order to eliminate floating cells. The cells were again treated with the previous conditions.

[0168] Pictures were taken immediately (x4 magnification, PICO, Molecular Devices) and after 8 hours and after 24 hours of incubation at 37 °C with 5% CO2.

[0169] Quantification of Scratched Area and Cell Migration

[0170] The area of scratch was measured on the pictures using the Imaged software (Java 1 .6).

[0171] Statistical Analysis

[0172] A Shapiro- Wilk normality test was performed to evaluate whether the data follow the Gaussian Law. The results did not follow the Gaussian Law.

[0173] As a consequence, a non-parametric statistical analysis was performed by Kruskal- Wall is ANOVA followed by Mann Whitney U test. Results were considered significant with p<0.05 with *, p<0.01 with ** and p<0.001 with ***.

[0174] Results

[0175] The presented results are normalized relative to the untreated control and are shown in the following table: The pre-treatment with the positive reference (Heparin-binding EGF-like growth factor = HB-EGF at 1 ng / ml) increased the percentage of closing by +68% after 8 hours in comparison to the untreated condition. These results confirmed the responsiveness of the model.

[0176] In the same conditions, the blue pigment of the invention at 0.05 mg / ml significantly increased the keratinocytes migration by up to +60%* (after 8 hours).

[0177] These results show that the pigment of the invention at 0.05 mg / ml exhibits a wound healing activity under these experimental conditions.

[0178] Example 9: Evaluation of Spreadability in Cosmetic Formulation

[0179] Powder samples 1 , 2, 5, and 6 of example 4 were incorporated into a lipstick formulation as described in example 6 at a concentration of 10%.

[0180] The colorimetric properties of the resulting lipstick compositions were measured using a Spectrocolorimeter Konica-Minolta 3600d (h, C*), and the coverage (= Contrast Ratio Opacity; Cr; Cr(%) =((Y black*100 / Y white), with Y being the luminance or brightness).

[0181] As can be seen from the above, the coverage of the formulations increased drastically with micronization. And also the chroma increased.

[0182] Figure 2 shows photographs of the above formulations spread out on contrast cards.

[0183] As can be seen from these photographs, the spreadability of the formulations containing micronized pigments (2* and 6*) was much better than that of the other two formulations (1* and 5*), and the latter two even contained some visible particles.

[0184] Thus, overall, the pigments of the present invention exhibit a significantly better coverage and also provide the desired blue color.

Claims

Claims1. Pigment for use in cosmetic applications, comprising micronized Gardenia Blue and kaolinite clay.

2. Pigment according to claim 1 , wherein the Gardenia Blue is obtained from a Gardenia florida L. fruit extract, preferably by hydrolysis and treatment with an amine, in particular with a primary amine.

3. Pigment according to claim 2, wherein the hydrolysis is an enzymatic hydrolysis.

4. Pigment according to claim 2 or 3, wherein the amine is selected from an amino acid, an amino acid salt, a peptide, a protein, and mixtures thereof, more preferably wherein the amine is selected from glutamine, a glutamate salt, and mixtures thereof.

5. Pigment according to any one of claims 2 to 4, wherein the Gardenia florida L. fruit extract comprises from about 2 wt% to about 12 wt% of geniposide, more preferably from about 4 wt% to about 10 wt% of geniposide, and most preferably from about 6 wt% to about 8 wt% of geniposide, based on the dry weight of the Gardenia florida L. fruit extract.

6. Pigment according to any one of claims 1 to 5, wherein the Gardenia Blue comprises less than about 12 ppm of genipin, more preferably less than about 10 ppm of genipin, still more preferably less than about 5 ppm of genipin, and most preferably wherein no genipin is detected.

7. Pigment according to any one of claims 1 to 6, wherein the micronized Gardenia Blue has a volume median particle size (Dv50) of from about 1 .0 pm to about 7.5 pm, more preferably of from about 1.5 pm to about 6.0 pm, and most preferably from about 2.0 pm to about 5.0 pm.

8. Pigment according to any one of claims 1 to 7, wherein the micronized Gardenia Blue has a volume particle size 90% percentile (Dv90) of from about 3.0 pm to about 20.0 pm, more preferably of from about 4.0 pm to about 17.0 pm, and most preferably from about 5.0 pm to about 15.0 pm.

9. Pigment according to any one of claims 1 to 8, wherein the weight ratio of micronized Gardenia Blue to kaolinite clay is from about 1 :4 to about 1 :1 , more preferably from about 1 :3 to about 1 :2, and most preferably about 1 :2.5.

10. Pigment according to any one of claims 1 to 9, further comprising a polysaccharide, more preferably maltodextrin.11 . Pigment according to claim 10, comprising from about 5 wt% to about 20 wt% of micronized Gardenia Blue, from about 20 wt% to about 40 wt% of kaolinite clay, and from about 50 wt% to about 65 wt% of maltodextrin; more preferably comprising from about 10 wt% to about15 wt% of micronized Gardenia Blue, from about 25 wt% to about 35 wt% of kaolinite clay, and from about 55 wt% to about 60 wt% of maltodextrin, for example about 12 wt% of micronized Gardenia Blue, about 30 wt% of kaolinite clay, and about 58 wt% of maltodextrin.

12. Pigment according to any one of claims 1 to 11 , wherein at least all solid components of the pigment are micronized.

13. Cosmetic composition comprising the pigment according to any one of claims 1 to 12 and a cosmetically acceptable excipient, in particular a cosmetically acceptable excipient comprising castor oil.

14. Cosmetic composition according to claim 13, which is a make-up composition, and in particular a lipstick composition, an eye shadow composition or a compacted powder composition.

15. Cosmetic composition according to claim 13, which is a skin care composition.

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