Lactic acid fermented product of hybrid tea rose petal extract

Abstract

To provide means for solving skin irritation, inflammation, and pain caused by cytotoxicity when applying a skin external preparation containing an extract of rose petals.SOLUTION: The problem is solved by using a skin external preparation containing a fermented product obtained by fermenting a hybrid tea rose petal extract with lactic acid bacteria.EFFECT: According to the present invention, a fermented product and a skin external preparation that maintain the effects of a rose petal extract before fermentation and that have low cytotoxicity are provided. This makes it possible to incorporate a high proportion of a lactic acid bacteria-fermented product of a rose petal extract that maintains the effects of the rose petal extract, and it is possible to exhibit a higher effect than the conventional one. Furthermore, even people with sensitive skin and infants can use a formulation containing a lactic acid bacteria-fermented product of a rose petal extract that maintains the effects of the rose petal extract.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a fermented product obtained by fermenting a petal extract of a hybrid tea rose with lactic acid bacteria, and more particularly to a fermented product having lower cytotoxicity than the petal extract and a skin external preparation containing the fermented product.

Background Art

[0002] Rose is a general term for plants classified in the genus Rosa of the Rosaceae family, and there are various species. As one classification, there are wild species and cultivated varieties born from their crossbreeding. As wild species, Rosa centifolia, Rosa damask, Rosa canina, etc. are known, and as cultivated varieties, mainly Rosa hybrida is known. Although it is difficult to genetically classify the currently said tens of thousands of cultivated varieties of roses, cultivated varieties having a continuous blooming property derived from the crossbreed of a hybrid perpetual rose originating from Rosa gallica and Rosa chinensis, a wild species native to China having a continuous blooming property, are collectively called hybrid tea roses. In recent years, many roses used for ornamental purposes are hybrid tea roses (Non-Patent Document 1).

[0003] Hybrid tea roses are not only used for ornamental purposes but also used as fragrant water to enjoy their characteristic fragrance. The petals of hybrid tea roses have a different fragrance from those of wild species such as Rosa gallica, and it is known that 1,3-dimethoxy-5-methylbenzene is commonly contained as a characteristic fragrance component (Non-Patent Document 2). Further, since it is known that this 1,3-dimethoxy-5-methylbenzene has a sedative effect (Patent Document 1), it is considered that the fragrance of the petals of hybrid tea rose varieties in general has a relaxing effect. As described above, since there are components commonly contained in hybrid tea roses in general, it is recognized as a group derived from a common cross parent.

[0004] Beyond its fragrance, rose petal extracts have long been reported to have beneficial effects on the human body. For example, Patent Document 2 describes that extracts of Western rose petals have the effect of suppressing the production of carbonyl proteins. Patent Document 3 describes that extracts of rose petals are effective as reactive oxygen species scavengers exhibiting mucopolysaccharide fragmentation inhibitory effects and superoxide dismutase-like activity. Furthermore, Patent Document 4 describes that extracts of Western roses are effective as a topical skin whitening agent. Patent Document 5 describes that extracts of hybrid tea roses are effective as elastase activity inhibitors, antioxidants, and collagenase activity inhibitors.

[0005] On the other hand, while the beneficial effects of rose petal extract have been confirmed as described above, when used as a topical agent on the skin, incorporating high concentrations of rose petal extract into the agent increases its cytotoxicity, resulting in skin irritation and pain. Therefore, high concentrations can cause skin irritation, inflammation, and pain due to cytotoxicity, making it unsuitable for people with sensitive skin or infants with insufficient barrier function who are easily irritated. Typically, the above problems occur when using topical agents containing 0.01% by mass or more of rose petal extract, calculated as evaporation residue. Thus, there was a dilemma in that while it was desirable to use high concentrations to fully exert the effects of rose petal extract, high concentrations resulted in cytotoxicity, and resolving this issue was a challenge.

[0006] Lactic acid bacteria are a general term for bacteria that produce lactic acid through metabolism and contribute to the fermentation of foods such as yogurt, pickles, and lactic acid bacteria beverages. They are known to break down glucose to produce lactic acid, which lowers the pH of the environment, thereby suppressing the growth of microorganisms that cause spoilage and food poisoning, and enabling the long-term preservation of food. In addition to their use in food, Patent Document 6 describes that fermenting Western rose petals or extracts with lactic acid bacteria enhances anti-allergic and whitening effects. On the other hand, a problem with lactic acid bacteria fermented products is that the metabolites of lactic acid bacteria are known to exhibit cytotoxicity (Non-Patent Document 3), so the use of lactic acid bacteria fermented Western roses has been limited because the cytotoxicity is higher than before fermentation. Furthermore, there are no known examples of lactic acid bacteria fermentation of hybrid tea roses, and although the anti-allergic effect of the aforementioned Western rose petal fermented product is known, its effect of reducing cytotoxicity, which has a different mechanism of action than allergies, was completely unknown. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Japanese Patent Application Publication No. 06-172781 [Patent Document 2] Japanese Patent Publication No. 2009-298726 [Patent Document 3] Japanese Patent Application Publication No. 7-309770 [Patent Document 4] Japanese Patent Publication No. 2002-029959 [Patent Document 5] Japanese Patent Publication No. 2011-236147 [Patent Document 6] Japanese Patent Publication No. 2010-270152 [Non-patent literature]

[0008] [Non-Patent Document 1] Yotaro Tsukamoto, Encyclopedia of Horticultural Plants, 1994. [Non-Patent Document 2] Perfumer & Flavarist Volume12,1987 [Non-Patent Document 3] BioMed Research International.D-Lactic Acid as a Metabolite: Toxicology, Diagnosis, and Detection 1-9,2020(2) [Overview of the project] [Problems that the invention aims to solve]

[0009] The objective is to provide a means to resolve skin irritation, inflammation, and pain caused by cytotoxicity resulting from the application of topical skin preparations containing rose petal extracts. [Means for solving the problem]

[0010] The inventors of this invention conducted diligent research to solve the above problems and found that they solved the problems by using a fermented product obtained by fermenting a petal extract of hybrid tea rose with lactic acid bacteria. [Effects of the Invention]

[0011] The present invention provides a fermented product and a topical skin preparation that maintain the effects of the rose petal extract before fermentation while having low cytotoxicity. This makes it possible to incorporate a high concentration of the lactic acid fermented product of rose petal extract while maintaining the effects of the rose petal extract, thereby achieving a higher efficacy than conventional products. Furthermore, it makes it possible for people with sensitive skin and infants to use formulations containing the lactic acid fermented product of rose petal extract while maintaining the effects of the rose petal extract. [Modes for carrying out the invention]

[0012] The present invention will be described in detail below.

[0013] The hybrid tea roses used in the present invention are cultivated varieties of roses that bloom repeatedly throughout the seasons and are not particularly limited to any rose classified as a hybrid tea rose (Rosa hybrida). For example, pink varieties include Lady Luck, Karina, Charlotte Armstrong, Confidence, Dainty Beth, First Prize, Michelle Meilland, Momoyama, Ophelia, Royal Highness, and Sonia; yellow varieties include Keihakitami, Golden Heart, Earl's Menol Gold, Amatsu Otome, Arlene Francis, Golden Masterpiece, Golden Lovechair, Golden Scepter, Helmut Schmidt, and Peace; and red varieties include Papa Meilland, Oklahoma, Charles Mallerin, Christian Dior, Chrysler Imperial, Crimson Glory, Ina Harkness, Grand Masterpiece, Happiness, and Jo The following varieties can be used: Waterer, Sid Deville, Baccara, Duftwolke, Lady Rose, Suzaku, Summer Holiday, Superstar, the white varieties Shosetsu, Blanche Malin, Honor, Message, Pascali, Virgo, White Christmas, White Masterpiece, Caramel Antique, the purple varieties Shiko, Black Tea, Blue Moon, Gray Pearl, Lady Ex, Madame Violet, Sterling Silver, the multi-colored varieties Double Delight, Black Gold, Garden Party, Kordes Perfecta, Seika, Colorama, Königindelrosen, Love, Modern Times, Piccadilly, Tzigane, etc. Due to their availability and industrial advantages such as the harvestable weight of petals, Lady Luck, Keihakitami, Golden Heart, Papa Meilland, Oklahoma, Shosetsu, Shiko, and Double Delight are particularly preferable. Hybrid varieties produced by crossing the above varieties can also be used in the same way. The name of the hybrid tea rose of the present invention may have multiple names, such as the trade name or product name given when it is distributed in the market, the variety registration name registered under the Plant Variety Protection Act, or the trademark registered under the Trademark Act. However, as long as it is classified as a hybrid tea rose, it is not limited to the names exemplified above.

[0014] The fermented product of the present invention is a fermented product (hereinafter referred to as hybrid tea rose fermented product) obtained by fermenting a petal extract of hybrid tea rose (hereinafter referred to as hybrid tea rose extract) with lactic acid bacteria. However, for industrial production, operational reasons, etc., it is acceptable for parts other than petals (for example, calyx, filaments, anthers, stigma, style, ovary, ovule, pedicel, flower stem) to be included in the preparation process.

[0015] The method for preparing hybrid tea rose extract for fermentation by lactic acid bacteria is not particularly limited, but for example, it can be prepared by extracting hybrid tea rose petals with a solvent. Alternatively, the liquid obtained by extracting the water contained in the hybrid tea rose petals themselves using methods such as pressing and vacuum drying can also be used as the hybrid tea rose extract.

[0016] The solvent for obtaining the hybrid tea rose extract is not particularly limited. For example, water (such as purified water), aqueous glucose solution, aqueous solution of inorganic salts (such as sodium chloride, potassium chloride, magnesium chloride, ammonium carbonate, etc.), buffer solutions (such as phosphate buffer, acetate buffer, Tris-hydrochloric acid buffer, carbonate buffer, borate buffer, etc.), aqueous solutions of inorganic acids (such as hydrochloric acid, carbonic acid, sulfuric acid, nitric acid, phosphoric acid, etc.), aqueous solutions of organic acids (such as acetic acid, citric acid, lactic acid, succinic acid, ascorbic acid, fumaric acid, malic acid, etc.), aqueous solutions of surfactants (such as saponin, lecithin, etc.), lower alcohols (such as methanol, ethanol, propanol, butanol, etc.), ketones (such as acetone, ethyl methyl ketone, etc.), hydrocarbons (such as propane, butane, hexane, cyclohexane, etc.), ethers (such as diethyl ether, etc.), glycols (such as ethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, etc.), halogenated hydrocarbons (such as dichloromethane, 1,1,1,2-tetrafluoroethane, 1,1,2-trichloroethene, etc.), acetic acid esters (such as ethyl acetate, methyl acetate, etc.) or a mixture thereof can be used. The mixing ratio (weight ratio) of the petals of the hybrid tea rose to the solvent is generally in the range of 1:1 to 1:1000, preferably 1:5 to 1:100, more preferably 1:10 to 1:50 in terms of the dry weight conversion of the petals.

[0017] The extraction process for obtaining hybrid tea rose extract is not particularly limited, but can be carried out, for example, by the following method. First, the petals of the hybrid tea rose to be extracted are immersed or dispersed in the solvent. In this case, the petals may be used fresh, or they may be dried or semi-dried beforehand. The shape is not particularly limited, but they may be used as they are after being collected, or they may be finely chopped or pulverized. The extraction process only requires immersing or dispersing the hybrid tea rose petals in the solvent, and there is no need to set a specific extraction time, but an appropriate extraction time may be set to prevent the petals from rotting. The extraction temperature is not particularly limited, but it may be set as appropriate, such as room temperature (20°C to 30°C) or heated (30°C to 100°C). Before subjecting the obtained hybrid tea rose extract to the fermentation process, it is preferable to separate the liquid phase by solid-liquid separation means such as filtration or centrifugation to remove the immersed or dispersed petals, but this solid-liquid separation step may be omitted if it does not hinder the fermentation process. The hybrid tea rose extract may be used directly for fermentation after preparation, or it may be purified according to conventional methods before use, or it may be diluted or concentrated to an appropriate concentration if necessary. In some cases, the liquid phase after solid-liquid separation may be solidified according to conventional methods such as spray drying or freeze-drying, and then further pulverized into a powder if necessary before use.

[0018] The hybrid tea rose extract prepared by the above method is preferably sterilized as necessary before being subjected to the fermentation process to remove contaminating bacteria that may interfere with fermentation. In this case, as a method for removing contaminating bacteria, a method may be used in which the petals to be used are washed and sterilized in advance with sterilizing ethanol or the like and then extracted with a sterile solvent, or a method may be used in which the petals are extracted with a solvent, and after obtaining an extract, the extract is sterilized by heating. As the heat sterilization method, an autoclave sterilization method in which heating is performed at 105 to 121°C for 10 to 20 minutes or an intermittent sterilization method in which holding at 80 to 90°C for 60 to 120 minutes is repeated once a day for 2 to 3 days is generally used. However, this sterilization step is not an essential step. If there is no problem in the subsequent fermentation step, this sterilization step may be omitted.

[0019] The lactic acid bacteria used for fermentation in the present invention are not particularly limited, but from the viewpoint of versatility, Lactobacillus plantarum is preferred. It is preferable to pre-culture the bacteria in a basic culture medium before subjecting them to the fermentation process. Culturing at this stage can be done in the usual manner as long as the lactic acid bacteria can grow. For the basic culture medium for lactic acid bacteria, it is sufficient to include a minimum amount of carbon, nitrogen, and phosphorus sources. However, it is preferable to use a medium developed to promote good overall growth of lactic acid bacteria, such as 310-9 medium or MRS medium. (310-9 medium composition: peptone 2.5g, beef extract 2.5g, yeast extract 5g, glucose 20g, Tween80 1g, K2HPO4 2g, sodium acetate 5g, ammonium dihydrogen citrate 2g, MgSO4·7H2O 0.2g, MnSO4·nH2O 0.05g, purified water 1L) (MRS medium composition: peptone 10g, beef extract 10g, yeast extract 5g, glucose 20g, Tween80 1g, K2HPO4 2g, sodium acetate 5g, ammonium dihydrogen citrate 2g, MgSO4·7H2O 0.2g, MnSO4·nH2O (0.05g, 1L purified water). Even with a culture medium composition other than those mentioned above, any substance that lactic acid bacteria can utilize and proliferate is acceptable for application to the present invention.Furthermore, the basic culture medium may contain aqueous solutions of inorganic salts (sodium chloride, potassium chloride, magnesium chloride, ammonium carbonate, etc.) or buffers (phosphate buffer, acetate buffer, Tris-HCl buffer, carbonate buffer, borate buffer, etc.), aqueous solutions of inorganic acids (hydrochloric acid, carbonic acid, sulfuric acid, nitric acid, phosphoric acid, etc.), aqueous solutions of organic acids (acetic acid, citric acid, lactic acid, succinic acid, ascorbic acid, fumaric acid, malic acid, etc.), aqueous solutions of surfactants (saponins, lecithin, etc.), lower alcohols (methanol, ethanol, propanol, butanol, etc.), and ketones (acetone, ethanol, etc.). Substances such as methyl ketone, hydrocarbons (propane, butane, hexane, cyclohexane, etc.), ethers (diethyl ether, etc.), glycols (ethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, etc.), halogenated hydrocarbons (dichloromethane, 1,1,1,2-tetrafluoroethane, 1,1,2-trichloroethene, etc.), acetate esters (ethyl acetate, methyl acetate, etc.), or mixtures thereof can be added, but if added, it is preferable to add them in a range that does not affect the growth of lactic acid bacteria. Furthermore, in the subsequent fermentation process, the above various components may be included in the solvent at concentrations that do not affect the fermentation of lactic acid bacteria.

[0020] The fermentation process for obtaining a hybrid tea rose ferment is not particularly limited, but the hybrid tea rose extract prepared by the above method can be inoculated with lactic acid bacteria and cultured for fermentation. The amount of lactic acid bacteria inoculated is not particularly limited, but preferably 10% of the extract. 5 ~10 10The concentration is in the range of cells / mL. The fermentation temperature is not particularly limited, but is generally in the range of 5 to 50°C, preferably in the range of 25 to 40°C, which is the optimal temperature for lactic acid bacteria growth. The number of fermentation days is not particularly limited, but is generally in the range of 1 to 15 days, preferably 3 to 10 days, at the optimal temperature. Fermentation culture can be carried out by standing, but shaking culture or aerated culture can also be performed to shorten the fermentation time. Once the above fermentation process is completed, it is preferable to heat sterilize the fermented culture solution that has gone through the fermentation process under the above conditions by heating at 80 to 120°C for about 15 to 120 minutes in order to stop the fermentation. The fermented culture solution after sterilization can be used as is, or the liquid phase can be separated by solid-liquid separation means such as filtration or centrifugation to obtain the fermented product.

[0021] The hybrid tea rose fermented product may be used as is after preparation, or it may be purified according to conventional methods before use, or it may be diluted or concentrated to an appropriate concentration if necessary. In some cases, the liquid phase after solid-liquid separation may be solidified according to conventional methods such as spray drying or freeze-drying, and then further pulverized into a powder if necessary before use.

[0022] When using hybrid tea rose ferment as a topical skin preparation according to the present invention, the hybrid tea rose ferment may be used as is, or it may be mixed with a general base. The final form can be any form, such as liquid, emulsion, gel, solid, powder, or granules, and any optional ingredients used in topical skin preparations can be added as needed, as long as the effect is not impaired. Examples of such optional ingredients include oils, surfactants, powders, colorants, water, alcohols, thickeners, chelating agents, silicones, antioxidants, UV absorbers, humectants, preservatives, fragrances, various medicinal ingredients, pH adjusters, and neutralizing agents. Furthermore, as the final form, examples of skin preparations include basic cosmetics such as emulsions, creams, lotions, essences, gels, packs, and facial cleansers; makeup cosmetics such as lipsticks, foundations, liquid foundations, and pressed makeup powders; cleansing cosmetics such as facial cleansers, body shampoos, and soaps; and bath additives, but are not limited to these. The amount of hybrid tea rose ferment in each preparation can be adjusted as appropriate according to the desired effect, but the optimal range is 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass, when converted to evaporation residue.

[0023] Cytotoxicity is damage to cells and is not particularly limited, but can be evaluated by factors such as cell death, respiration, metabolism, cell proliferation rate, and intracellular reactive oxygen species concentration. In this application, the determination of whether or not a substance is low in cytotoxicity is made by a relative evaluation of cytotoxicity before and after lactic acid fermentation treatment. That is, when comparing two substances, if the cytotoxicity of one substance is lower than that of the other substance after lactic acid fermentation treatment, it is determined to be low in cytotoxicity. For example, when comparing an extract with its fermented product (after fermentation treatment), if the cytotoxicity of the fermented product is lower than that of the extract, the fermented product can be determined to be less cytotoxic than the extract. [Examples]

[0024] The following describes examples of efficacy tests of the hybrid tea rose ferment in the present invention. Furthermore, examples of application formulations for topical skin preparations using the hybrid tea rose ferment will be described, but the invention is not limited to the examples described herein.

[0025] <Preparation of lactic acid bacteria to be inoculated into the extract 1> Lactobacillus plantarum was used as the lactic acid bacterium. One loopful of the lactic acid bacterium strain was inoculated into 50 mL of 310-9 medium and incubated at 30°C for 3 days to obtain the lactic acid bacterium culture solution. (310-9 medium composition: peptone 2.5 g, beef extract 2.5 g, yeast extract 5 g, glucose 20 g, Tween 80 1 g, K2HPO4 2 g, sodium acetate 5 g, ammonium dihydrogen citrate 2 g, MgSO4·7H2O 0.2 g, MnSO4·nH2O 0.05 g, purified water 1 L)

[0026] <Preparation of lactic acid bacteria to be inoculated into the extract 2> Lactobacillus plantarum was used as the lactic acid bacterium. One loopful of the lactic acid bacterium strain was inoculated into 30 mL of MRS medium and incubated at 30°C for 3 days to obtain the lactic acid bacterium culture solution. (MRS medium composition: peptone 10 g, beef extract 10 g, yeast extract 5 g, glucose 20 g, Tween 80 1 g, K2HPO 4 2 g, sodium acetate 5 g, ammonium dihydrogen citrate 2 g, MgSO4·7H2O 0.2 g, MnSO4·nH2O 0.05 g, purified water 1 L)

[0027] <Preparation of extracts and fermented products 1> 5g each of dried hybrid tea rose petals (Lady Luck, Golden Heart, Papa Meilland, Oklahoma, and Purple Fragrance) were added to 50mL of a 1% glucose aqueous solution and sterilized to obtain hybrid tea rose extract. 1mL of the lactic acid bacteria culture solution prepared in

[0025] was added to the obtained extract and mixed well. The extract inoculated with lactic acid bacteria was cultured at 30°C for one week and then sterilized. After sterilization, the petals and solids were removed by filtration to obtain hybrid tea rose ferment. As a comparative control 1, a 1% glucose aqueous solution without petals was used as the sugar solution (considered as the extract when calculating the ratio of extract / fermented product before and after fermentation), 1mL of lactic acid bacteria culture solution was added to the sugar solution and mixed well to inoculate the extract with lactic acid bacteria, which was then cultured at 30°C for one week and sterilized to obtain the sugar ferment. As a second comparative study, a rose extract and rose ferment were prepared from dried petals of the European rose, a different species from the hybrid tea rose, using the same method as for the hybrid tea rose extract and ferment described above. As a third comparative study, a rice bran extract and rice bran ferment were prepared from rice bran, which is commonly used in lactic acid fermentation, using the same method as for the hybrid tea rose extract and ferment described above.

[0028] <Preparation of extracts and fermented products 2> 10 g each of pre-dried hybrid tea rose petals (Double Delight, Keihakitami, Shosetsu) were mixed with 100 mL of purified water and extracted at room temperature (20°C-30°C). The petals were then removed by filtration to obtain a hybrid tea rose extract. 5 mL of the lactic acid bacteria culture solution prepared in

[0026] was added to the obtained extract and mixed well. The extract inoculated with lactic acid bacteria was cultured at 30°C for 3 days, and then sterilized at 105°C for 15 minutes. After sterilization, the solids were removed by filtration to obtain a hybrid tea rose ferment.

[0029] <Cytotoxicity Confirmation Test> The method for confirming the cytotoxicity of the fermented product prepared by the method described above is shown below. Cytotoxicity can be evaluated by a cell proliferation test using a WST assay reagent, which allows for the quantification of living cells by measuring the absorbance of the formazan dye produced by the metabolism of living cells at 450 nm. This test is also used for safety evaluation of topical skin preparations. A higher cell proliferation rate indicates lower cytotoxicity, and since cytotoxicity is highly correlated with skin irritation, it is known to be an indicator for evaluating the safety of a sample on the skin.

[0030] Human epidermal keratinocyte-derived cells 1 × 10 5 The cells were dispersed in Dulbecco's Modified Eagle Medium (DMEM) with 5% Fetal Bovine Serum (FBS) to a concentration of cells / mL, and 0.1 mL was seeded into a 96-well plate (final concentration 1.0 × 10⁶). 4 Cells / well). Cultured at 37°C, 5% CO2 for 24 hours. Remove the culture medium and wash with PBS(-). Add each sample (each extract, ferment) prepared in DMEM to a final concentration of 500 ppm. Cultured at 37°C, 5% CO2 for 72 hours, then remove the culture medium containing the samples. Wash with PBS(-). Add 0.2 mL of DMEM containing 9% WST assay reagent (Cell Counting Kit, DOJINDO) to each cell. Incubate at 37°C, 5% CO2 for 20 minutes. Divide each cell into a 96-well plate (0.1 mL). Measure the absorbance at 450 nm using a microplate reader.

[0031] <Data Analysis> The ratio of cell proliferation between each extract and the fermented product was calculated as the pre- and post-fermentation (extract / fermented product) ratio using the following formula.

[0032]

number

[0033] Table 1 shows the calculated cell proliferation rate before and after fermentation (extract / fermented product) ratio. HT in the table is an abbreviation for Hybrid Tea Rose. In all rose petals classified as Hybrid Tea Roses shown in Examples 1 to 8, the cell proliferation rate before and after fermentation (extract / fermented product) ratio was lower than 1, meaning that the Hybrid Tea Rose fermented product had a higher cell proliferation rate and therefore lower cytotoxicity compared to the Hybrid Tea Rose extract. On the other hand, in the comparative examples of sugar solution, Western rose, and rice bran, the cell proliferation rate before and after fermentation (extract / fermented product) ratio was higher than 1, meaning that the fermented product had higher cytotoxicity compared to the extract. Hybrid Tea Roses have different pigment components depending on the color system of the petals, but the above trend was observed in all color systems. However, in fermented products other than Hybrid Tea Rose petals and fermented products of Western roses of a different species, the cytotoxicity was higher than that of their extracts, confirming that this effect is specific to Hybrid Tea Rose extracts within the Rosa genus. Furthermore, the known effects of rose petal extract—inhibitory effect on carbonylation proteins, inhibitory effect on mucopolysaccharide fragmentation, reactive oxygen species scavenging effect exhibiting superoxide dismutase-like activity, skin whitening effect, elastase activity inhibitory effect, and collagenase activity inhibitory effect—were maintained unchanged before and after fermentation.

[0034] [Table 1]

[0035] Next, examples of formulations for topical skin preparations containing the hybrid tea rose ferment of the present invention are shown, but the present invention is not limited thereto. In the following, all parts refer to parts by mass, and all percentages refer to mass percent. The hybrid tea rose ferment shown in each of the following formulation examples was prepared by the method described in

[0027] or

[0028] , and its amount is shown as mass percent converted to evaporation residue. In each of the formulation examples, no symptoms such as skin irritation, inflammation, or pain were observed when using topical skin preparations containing the hybrid tea rose ferment in users who had experienced irritation with conventional topical skin preparations containing hybrid tea rose extract or users who considered themselves to have sensitive skin. On the other hand, the effectiveness as a topical skin preparation remained unchanged before and after fermentation.

[0036] <Examples of prescriptions for topical skin preparations> (Formulation Example 1) Cosmetic cream (by mass %) a) Beeswax...2.0 b) Stearyl alcohol... 5.0 c) Stearic acid... 8.0 d) Squalane...10.0 e) Self-emulsifying glyceryl monostearate...3.0 f) Polyoxyethylene cetyl ether (20 E.O.) ··· 1.0 g) Fermented hybrid tea rose (Lady Luck) (prepared with

[0027] ) ... 0.1 h) 1,3-Butylene glycol...5.0 i) Potassium hydroxide...0.3 j) Preservatives and antioxidants... appropriate amount k) Purified water...remainder Total 100.0 Manufacturing method Heat and dissolve ingredients a) through f), maintaining the temperature at 80°C. Heat and dissolve ingredients h) through k), maintaining the temperature at 80°C, add them to a) through f), emulsify, and cool to 40°C while stirring. Then add g), stir, and dissolve uniformly.

[0037] (Formulation Example 2) Emulsion (mass %) a) Beeswax...0.5 b) Vaseline...2.0 c) Squalane... 8.0 d) Sorbitan sesquioleate...0.8 e) Polyoxyethylene oleyl ether (20 E.O.) ... 1.2 f) Fermented hybrid tea rose (Golden Heart) (prepared with

[0027] ) ... 0.05 g) 1,3-Butylene glycol ···7.0 h) Carboxyvinyl polymer...0.2 i) Potassium hydroxide...0.1 j) Purified water...remainder k) Preservatives and antioxidants... appropriate amount l) Ethanol ···7.0 Total 100.0 Manufacturing method Heat and dissolve ingredients a) through e) and maintain the temperature at 80°C. Heat and dissolve ingredients g) through k) and maintain the temperature at 80°C, add them to a) through e) and emulsify, then cool while stirring to 50°C. At 50°C, add f) and l), stir and cool to 40°C.

[0038] (Formulation Example 3) Lotion (mass %) a) Hybrid tea rose (Keihakitami) ferment (prepared with

[0028] ) ... 0.01 b) Glycerin...5.0 c) Polyoxyethylene sorbitan monolaurate (20 E.O.) ···1.0 d) Ethanol ···6.0 e) Fragrance...appropriate amount f) Preservatives and antioxidants... appropriate amount g) Purified water...remainder Total 100.0 Manufacturing method Mix ingredients a) through g) and dissolve them uniformly.

[0039] (Formulation Example 4) Facial Cleanser (by mass) a) Stearic acid...12.0 b) Myristic acid...14.0 c) Lauric acid...5.0 d) Jojoba oil...3.0 e) Glycerin...10.0 f) Sorbitol...15.0 g) 1,3-Butylene glycol ···10.0 h) POE(20) Glycerol Monostearate Ester ···2.0 i) Potassium hydroxide...5.0 j) Purified water...remainder k) Chelating agent... appropriate amount l)Fragrance...appropriate amount m) Hybrid Tea Rose (Oklahoma) Ferment (prepared with

[0027] ) ... 0.01 Total 100.0 Manufacturing method Heat and dissolve a) through h) and maintain at 70°C. Dissolve i) in j), then add to a) through h) and saponify. Then add k) and l) and cool while stirring. Add m) at 50°C, stir to 40°C, and cool.

[0040] (Formulation Example 5) Essence (by mass) a) Hybrid tea rose (purple fragrance) ferment (prepared with

[0027] ) ... 0.1 b) Carboxyvinyl polymer...0.05 c) L-arginine... appropriate amount d) Glycerin ··· 5.0 e) Polyoxyethylene sorbitan monolaurate (20 E.O.) ···1.0 f) Ethanol...6.0 g)Fragrance...appropriate amount h) Preservatives and antioxidants... appropriate amount i) Purified water...remainder Total 100.0 Manufacturing method After dispersing b) in a portion of i), add c) to adjust the pH to 6.5. Then mix a) through i) until uniformly dissolved.

[0041] (Formulation example 6) Gel (mass %) a) Hybrid Tea Rose (Double Delight) Ferment (prepared with

[0028] ) ... 0.01 b) Carboxyvinyl polymer...0.5 c) Sodium hydroxide...0.05 d) Methyl parahydroxybenzoate...0.1 e) Cyclohexane-1,4-dicarboxylic acid bisethoxydiglycol...0.5 f) Polyglyceryl-10 (eicosanedioic acid / tetradecanediic acid)...0.5 g)PEG / PPG / Polybutylene Glycol-8 / 5 / 3 Glycerin···0.5 h) Polyoxyethylene (60 E.O.) hydrogenated castor oil... 0.1 i)Fragrance...appropriate amount j) Preservatives and antioxidants... appropriate amount k) Purified water...remainder Total 100.0 Manufacturing method After dispersing b) in a portion of k), add c). Then mix a) through k) until uniformly dissolved.

[0042] (Formulation example 7) Lipstick (mass %) a) Fermented hybrid tea rose (Papa Meilland) (prepared with

[0027] ) ... 0.0001 b) Candelilla Low... 9.0 c) Solid paraffin...8.0 d) Beeswax...5.0 e) Carnauba wax... 5.0 f) Lanillin···11.0 g) Castor oil... remaining portion h) Cetyl 2-ethylhexanoate...0.5 i) Isopropyl myristic acid ester...10.0 j) Titanium dioxide...5.0 k) Red No. 201...0.6 l) Red No. 202...1.0 m) Red No. 223...0.2 n)Fragrance...appropriate amount o) Preservatives and antioxidants... appropriate amount Total 100.0 Manufacturing method Disperse ingredients j) to m) into a portion of g) and process with a roller. Then mix ingredients a) to o), heat and melt, and then disperse uniformly. After dispersion, pour into a mold and cool rapidly to form sticks.

[0043] (Formulation Example 8) Powder Foundation (by mass %) a) Fermented hybrid tea rose (Shosetsu) (prepared with

[0028] ) ... 0.0001 b) Squalane...10.0 c) Sorbitan sesquioleate...3.5 d) Preservatives and antioxidants... appropriate amount e) Titanium dioxide...13.0 f) Sericite...25.0 g) Talc... Remaining portion h) Bengara (red iron oxide pigment)...0.8 i) Yellow iron oxide...2.5 j) Black iron oxide...0.1 Total 100.0 Manufacturing method Mix e) to j) and grind them in a grinder. Then add the mixed and dissolved a) to d), stir and mix, grind again, and press into a metal dish container.

[0044] (Formulation example 9) Bath additive (mass %) a) Hybrid tea rose (Keihakitami) ferment (prepared with

[0028] ) ... 0.005 b) Cyclohexane-1,4-dicarboxylic acid bisethoxydiglycol...0.2 c) Polyglyceryl-10 (eicosanedioic acid / tetradecanediic acid)...0.2 d) PEG / PPG / Polybutylene Glycol-8 / 5 / 3 Glycerin···0.2 e) Sodium bicarbonate...50.0 f) Sodium sulfate... remainder g)Fragrance...appropriate amount h) Preservatives and antioxidants... appropriate amount Total 100.0 Manufacturing method Mix a) through h) uniformly.

Claims

1. This is a lactic acid fermented product of petal extract from the petals of the hybrid tea rose Double Delight, The lactic acid bacteria fermented product is less cytotoxic than the petal extract before lactic acid bacteria fermentation.

2. A topical skin preparation containing the lactic acid bacteria fermented product described in claim 1.

Images