Skin external composition containing ginseng seed fermented oil with permeation enhancing effect and application of skin external composition

Abstract

The invention relates to a skin external composition containing ginseng seed fermented oil with a permeation enhancing effect and application of the skin external composition. The skin external composition comprises a whitening active ingredient and the ginseng seed fermented oil. Wherein the ginseng seed fermented oil can effectively promote skin permeability of whitening active ingredients, and further enhance whitening and skin care effects.

Description

Technical Field

[0001] The invention relates to the field of cosmetics, and in particular to a skin external composition containing ginseng seed fermented oil with penetration-promoting effect and application thereof. Background Art

[0002] Due to the skin's own barrier function, whitening active ingredients are difficult to penetrate into the basal layer of the skin and cannot directly act on melanocytes, greatly reducing the whitening efficiency. As a result, most whitening products such as glycyrrhizin, arbutin, ellagic acid, niacinamide, kojic acid, 4-butylresorcinol, hexylresorcinol, phenethylresorcinol, tranexamic acid, ascorbyl glucoside, ascorbyl tetraisopalmitate, 3-O-ethyl ascorbic acid, and magnesium ascorbyl phosphate have insufficient whitening efficiency.

[0003] In order to improve the whitening effect, a combination of ingredients that enhance penetration is currently used. For example, CN117838609B discloses a method for preparing cosmetics that improves transdermal absorption rate. Gallic acid and phosphatidylcholine are mixed to form a film by a thin film hydration method, and then placed in a phosphate buffer containing additives such as sodium glycodeoxycholate, isopropyl myristate and D-α-tocopherol polyethylene glycol 1000 succinate for hydration to form vesicles, and finally mixed with carboxymethyl cellulose sodium salt to prepare a gel matrix to obtain a transdermal absorption gel with a high transdermal absorption rate. The surface lipids of the prepared vesicles disrupt the orderly lipid structure of the skin, reduce its structural density, increase the fluidity of the stratum corneum, and change the lipids between skin cells, thereby facilitating the penetration of gallic acid. CN116459174A discloses a method for improving the permeability and retention rate of chemical components with skin care effects on the skin. In order to solve the problem that the effective ingredients of skin care components in cosmetics are difficult to enter the deep layer of the skin to play a role, the skin care components in cosmetics are compounded with alkyl glycosides to reduce the difficulty of the skin care components in cosmetics to enter the epidermal bottom layer and dermis. The alkyl glycosides can promote the transdermal absorption of the skin care components in cosmetics and increase the retention of the skin care components in the epidermis and dermis. CN114917146A is a transfer body and transfer body gel of a composition for solving the problem of skin aging in a multi-dimensional collaborative way. The effective ingredients are encapsulated in the transfer body vesicles, and the elastic deformation of the transfer body is used to easily penetrate the skin. The effective ingredients are delivered to the deep layer of the skin and then effectively play a role. It is made of phospholipids, cholesterol, edge activators, effective ingredients and water. CN106691888A discloses a high skin retention glabridin nanocomposition, the components and component ratios of which are closely related to promoting the penetration of active ingredients through the stratum corneum of the skin and improving skin retention performance, as well as improving the stability of the nanocomposition. The liposome structure composed of phospholipids and cholesterol in the composition can enter the interior of the stratum corneum cells of the skin, interact with keratin, reduce the density of the stratum corneum cells, and form lipid channels to promote the penetration of glycyrrhizin through the stratum corneum; isopropyl myristate is used as an oil phase component and also as a transdermal enhancer, which is easily absorbed by the skin; octyldodecanol is used as an O / W type emulsifier and has a transdermal enhancing effect, which can promote the transdermal absorption of glycyrrhizin; and octyldodecanol has good safety and is non-irritating to the skin; and vitamin E succinate polyethylene glycol ester is a water-soluble vitamin E derivative, which is composed of a hydrophilic polar polyethylene glycol segment and a lipophilic non-polar vitamin E succinate segment, and can inhibit P-glycoprotein in skin cells from excreting the skin care active ingredient glycyrrhizin into the blood, thereby increasing the retention time of glycyrrhizin in skin tissue.WO2023002872A1 discloses cosmetics for skin penetration, containing (a) an alkyl betaine-based amphoteric surfactant, (b) one or more selected from the group consisting of polyols, urea and urea derivatives, (c) a skin condition improving ingredient and (d) water. Component (a) is combined with component (b) as a solvent that acts as a penetration enhancer for the skin condition improving ingredient, dissolving the skin condition improving ingredient while appropriately reducing the barrier function of the stratum corneum, thereby improving the percutaneous absorbability of the skin condition improving ingredient. WO2024109386A1 discloses a whitening composition that promotes penetration and enhances synergy, comprising: a whitening ingredient; a Chinese medicine extract; and a polyol. The Chinese medicine extract is prepared from the following raw materials: Poria, Tribulus, Bletilla, Paeonia, Atractylodes, Bletilla and Dictamni, wherein the weight ratio of Poria: Tribulus: Bletilla: Paeonia: Atractylodes: Bletilla: Dictamni is 1: 1-4: 4-10: 8-10: 1-4: 1-4: 1-4, wherein the polyol is a mixture of butylene glycol and ethylhexylglycerol in a weight ratio of 9: 1. In a preferred embodiment, the whitening ingredient is selected from: niacinamide, ascorbyl glucoside or a combination thereof.

[0004] Therefore, the current research on the penetration enhancement of whitening ingredients is mainly on the research of formulation improvement and penetration enhancement ingredients, wherein formulation improvement also mainly adopts liposomes or vesicles or emulsions, but the preparation process of such preparations is complicated, the production cost is high, and the quality is difficult to control. For cosmetics containing more ingredients, this method is not suitable for industrialization. In the research on penetration enhancement ingredients, some specific surfactants are mainly used, but such surfactants are prone to cause skin sensitivity, irritation and other problems, or have limited penetration enhancement ability for whitening ingredients. Therefore, it is still necessary to develop a cosmetic composition that can nourish the skin, has a simple process, and can significantly promote the skin penetration of whitening ingredients. Summary of the invention

[0005] The present invention provides a skin external composition containing ginseng seed fermented oil with penetration-promoting effect and its use, wherein the ginseng seed fermented oil can effectively promote the skin permeability of whitening active ingredients, thereby enhancing the whitening and skin care effect. At the same time, the ginseng seed fermented oil has good skin care activity based on the ginseng seed oil itself, and the combination of the two further enhances the skin care effect.

[0006] The invention provides a skin external composition containing ginseng seed fermented oil with a penetration-promoting effect. The skin external composition is prepared from the following raw materials in the following weight proportions: 0.1-10 weight parts of a whitening active ingredient and 1-30 weight parts of ginseng seed fermented oil; the whitening active ingredient is selected from one or more of glabridin, arbutin, ellagic acid, nicotinamide, kojic acid, 4-butylresorcinol, hexylresorcinol, phenethylresorcinol, tranexamic acid, ascorbyl glucoside, ascorbyl tetraisopalmitate, 3-O-ethyl ascorbic acid, and magnesium ascorbyl phosphate; the ginseng seed fermented oil is an oil obtained by fermenting ginseng seed oil with yeast, and the ginseng seed fermented oil contains methyl palmitate (MEPA).

[0007] The ginseng seed fermented oil further comprises one or more of 2,4-di-tert-butylphenol (2,4-DTBP) and methyl stearate (MEST), and preferably further comprises 2,4-di-tert-butylphenol and methyl stearate.

[0008] The ginseng seed fermented oil contains 0.6-0.8 g / kg of 2,4-di-tert-butylphenol, and / or 1-1.5 g / kg of methyl stearate, and / or 0.5-0.7 g / kg of methyl stearate.

[0009] The whitening active ingredient is selected from glabridin or arbutin.

[0010] The yeast is selected from one or more of baker's yeast, feed yeast, brewer's yeast, Pichia pastoris, and Candida, preferably baker's yeast, Pichia pastoris, or Candida, and more preferably Candida bumblebee.

[0011] The ginseng seed fermented oil is prepared by the following method: after fermenting the ginseng seed oil with yeast, centrifuging the fermented liquid and taking the upper oily liquid. The ginseng seed oil can be obtained by pressing ginseng seeds, or supercritical extraction, or steam distillation, or ethanol extraction.

[0012] The present invention also provides a skin external preparation containing ginseng seed fermented oil with penetration-promoting effect, which comprises the skin external composition and skin-acceptable auxiliary materials.

[0013] The above-mentioned skin external preparation can be formulated into cream, ointment, gel, emulsion, facial mask, foam, lotion, essence, patch, hand pump type spray, stick, aerosol spray and wet tissue. It can be prepared into cosmetics, dermatological or pharmaceutical topical application products by methods commonly used in the art.

[0014] The skin external preparation of the present invention is an emulsion, cream, etc., comprising an aqueous phase and an oil phase. The aqueous phase is generally a mixture of water-soluble or dispersible components, which can be liquid or semisolid at room temperature. The aqueous phase can be selected from a mixture containing water or water and at least one polyol, and the polyol is generally selected from propylene glycol, butylene glycol, hexylene glycol, pentylene glycol, sorbitol, glycerol, panthenol or polyethylene glycol and mixtures thereof.

[0015] The oil phase of the present invention is generally a liquid oil-soluble or oil-dispersible component and / or an oily or waxy substance at room temperature, such as wax, semisolid, gum and mixtures thereof. Suitable oily substances are selected from: oils derived from animals, such as fully hydrogenated squalene; vegetable oils, such as corn oil, sunflower oil, soybean oil, castor oil, grape seed oil, avocado oil, jojoba oil, or liquid C4-10 fatty acid triglycerides, such as heptanoic acid or caprylic acid triglycerides, caprylic acid / capric acid triglycerides; or mineral oils, such as liquid paraffin and its derivatives, petrolatum; and synthetic oils, such as isopropyl myristate, isostearyl isostearate, 2-ethylhexyl palmitate, 2-octyldodecyl stearate; hydroxylated esters, such as octyldodecyl hydroxystearate. esters, such as propylene glycol dicaprylate, diethylene glycol diisononanoate and neopentyl glycol diheptanoate; fatty alcohols containing C12-26, such as oleyl alcohol, 2-butyloctanol, octyldodecanol, 2-hexyldecanol, 2-undecylpentadecane; or silicone oils, volatile or non-volatile linear or cyclic polymethylsiloxanes that are liquid or semisolid at room temperature, such as cyclopolydimethylsiloxane and polydimethylsiloxane.

[0016] The external preparation of the present invention may further contain a surfactant, and may further contain a film-forming polymer such as polyester, polyurethane, polyacrylic acid homopolymer or copolymer, hydrocarbon-based resin and / or silicone resin.

[0017] The topical preparation of the present invention may further comprise any component commonly used in the cosmetic field, such as preservatives, aqueous phase thickeners (extract biopolymers, synthetic polymers) and fat phase thickeners, antiallergic agents, antimicrobial agents, emollients, emulsifiers, exfoliants, insect repellents, antioxidants, chelating agents, colorants, depigmenting agents, film formers, pharmaceutically active agents, moisturizers, skin care agents, sunscreens, stabilizers, thickeners, humectants, lubricants, viscosity modifiers, fragrances, vitamins or any combination thereof.

[0018] The topical preparation of the present invention may also contain an additional particulate phase, which may be a pigment and / or pearlescent agent and / or filler used in a cosmetic composition. Suitable inorganic pigments include iron oxide and iron blue, titanium oxide, zirconium oxide and cerium oxide, and zinc oxide; suitable organic pigments include calcium, barium, strontium and aluminum lakes and carbon black. Suitable pearlescent agents include coated titanium oxide, iron oxide or natural pigment mica. Suitable fillers include silica, mica, kaolin, talc, nylon powder, polyethylene powder, Teflon, zinc stearate, boron nitride, copolymers.

[0019] The oil phase of the topical preparation of the present invention may contain one or more waxes, gums or mixtures thereof. Suitable waxes include carnauba wax, candelilla wax, beeswax, paraffin wax, microcrystalline wax, ceresin wax; synthetic waxes include polyethylene wax, silicone wax containing C16-45. Gums are generally polydimethylsiloxane or sodium carboxymethylcellulose or extracts, and semisolid substances are generally hydrocarbon-based compounds, such as lanolin and its derivatives.

[0020] The present invention further provides a skin topical cream containing ginseng seed fermented oil with a penetration-promoting effect, which includes, based on the mass of the cream, 0.1%-1% of a whitening active ingredient, 0.5%-10% of ginseng seed fermented oil, 3%-8% of stearyl alcohol, 6%-15% of glyceryl monostearate, 1%-10% of vegetable oil or mineral oil or triglyceride of C4-10 fatty acids, 8%-15% of a polyol, and the remainder of water; the whitening active ingredient is selected from glycyrrhizin or arbutin, the ginseng seed fermented oil is an oil obtained by fermenting ginseng seed oil with yeast, and the ginseng seed fermented oil contains methyl palmitate.

[0021] Preferably, 0.5% to 3% of ginseng seed fermented oil is included. The polyol includes glycerol and butylene glycol. The cream further includes carbomer and olive oil emulsifying wax.

[0022] The present invention also provides the use of the above-mentioned skin external composition or the above-mentioned skin external preparation or the above-mentioned skin external cream in preparing a whitening external preparation.

[0023] The present invention also provides the use of ginseng seed fermented oil in preparing an external composition for promoting skin penetration of whitening active ingredients. The ginseng seed fermented oil is an oil obtained by fermenting ginseng seed oil with yeast, and the ginseng seed fermented oil contains methyl palmitate MEPA. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 GC-MS chromatogram of unfermented ginseng seed oil

[0025] Figure 2 GC-MS chromatogram of fermented ginseng seed oil of Example 1 of the present invention

[0026] Figure 3 Curve of the effect of different addition amounts of fermented ginseng seed oil on the in vitro permeation of glabridin

[0027] Figure 4 Effects of MEST, 2,4-DTBP and MEPA on the penetration enhancement of glabridin DETAILED DESCRIPTION

[0028] Example 1

[0029] The preparation method of ginseng seed fermented oil in this embodiment comprises the following steps:

[0030] (1) Use an inoculation loop to pick up Candida albicans ATCC22214 from Bombus fasciatus and inoculate it into 100 mL of PDB liquid culture medium. Activate it at 28°C for 20 h to obtain yeast seed liquid. The concentration of the yeast seed liquid is about 4.5×10 7 cell / ml.

[0031] (2) Sterilize the ginseng seed oil separately in an autoclave at 115°C for 20 min.

[0032] (3) Sterilized ginseng seed oil and fermentation medium were mixed on a clean bench, and then 5% yeast suspension was added and incubated at 28°C and 150 r / min. -1 The culture was shaken for 48 to 96 hours under the following conditions: the fermentation medium included 60 g glucose, 3 g yeast powder, 1 g KH2PO4, 1 g K2HPO4, 0.5 g MgSO4, pH 6.5, and sterilized in an autoclave at 115°C for 20 min.

[0033] (4) After the fermentation is completed, the mixture is placed in an autoclave at 60°C for 1 hour, and then the temperature is continued to be raised for high temperature sterilization (115°C). The mixture is then cooled, separated, and centrifuged to obtain the upper clear and transparent oily liquid to obtain the fermented ginseng seed oil.

[0034] Among them, Candida albicans ATCC22214 was purchased from Ningbo Mingzhou Biotechnology Co., Ltd.

[0035] Example 2

[0036] (1) ATCC64110 Pichia pastoris was picked up with an inoculation loop and inoculated into 100 mL PDB liquid medium. The culture was activated at 28°C for 18 h to obtain yeast seed solution. The concentration of the yeast seed solution was approximately 4.0 × 10 7 cell / ml.

[0037] (2) Sterilize the ginseng seed oil separately in a high pressure sterilizer at 115°C for 20 to 30 minutes.

[0038] (3) Sterilized ginseng seed oil and fermentation medium were mixed on a clean bench, and then 4% yeast suspension was added and incubated at 26°C and 120 r / min. -1 The culture was shaken for 72 hours under the following conditions: the fermentation medium included 60 g glucose, 3 g yeast powder, 1 g KH2PO4, 1 g K2HPO4, 0.5 g MgSO4, pH 6.2, and sterilized in an autoclave at 115°C for 20 min.

[0039] (4) After the fermentation is completed, the mixture is placed in an autoclave at 80°C for 1 hour, and then the temperature is continued to be raised for high temperature sterilization (110°C). The mixture is then cooled, separated, and centrifuged to obtain the upper clear and transparent oily liquid to obtain the fermented ginseng seed oil.

[0040] Example 3

[0041] Preparation of cream containing glabridin and yeast-fermented ginseng seed oil

[0042] Dissolve glabridin powder in a small amount of 1,3-butylene glycol and set aside; weigh the raw materials of phase A according to Table 1, heat in a water bath at 75°C, and stir to dissolve; disperse carbomer in glycerol, stir evenly with a glass rod, then add other raw materials of phase B, heat in a water bath at 75°C, and stir to dissolve; after phases A and B are completely dissolved, pour phase A into phase B, stir evenly, and heat at 50000r·min -1 Homogenize and emulsify for 4 minutes, add phase C raw materials, stir evenly, and rotate at 4500r·min -1 Homogenize and emulsify for 5 minutes, cool to 50°C; add pre-dissolved glabridin, stir evenly, and discharge to obtain a cream containing glabridin and yeast-fermented ginseng seed oil. The raw materials in Table 1 are added in percentage by mass, ginseng seed oil is obtained by squeezing ginseng seeds, and fermented ginseng seed oil is the yeast-fermented ginseng seed oil prepared in Example 1.

[0043] Table 1 Recipe

[0044]

[0045] Example 4

[0046] Preparation of cream containing arbutin and yeast-fermented ginseng seed oil

[0047] Dissolve arbutin powder in a small amount of 1,2-hexanediol and set aside; weigh the raw materials of phase A according to Table 2, heat in a water bath at 70°C, and stir to dissolve; disperse carbomer in glycerol, stir evenly with a glass rod, then add other raw materials of phase B, heat in a water bath at 70°C, and stir to dissolve; after phases A and B are completely dissolved, pour phase A into phase B, stir evenly, and heat at 50000r·min -1 Homogenize and emulsify for 3 minutes, add phase C raw materials, stir evenly, and rotate at 5000r / min-1 Homogenize and emulsify for 5 minutes, cool to 50°C; add pre-dissolved arbutin, stir evenly, and discharge to obtain a cream containing arbutin and yeast-fermented ginseng seed oil. The raw materials in Table 2 are added in percentage by mass, the ginseng seed oil is obtained by squeezing ginseng seeds, and the fermented ginseng seed oil is the yeast-fermented ginseng seed oil prepared in Example 1.

[0048] Table 2 Recipe

[0049]

[0050] Comparative Example 1

[0051] The preparation method of the ginseng seed fermented oil of this comparative example comprises the following steps:

[0052] (1) Lactobacillus gasseri CGMCC 1.3396 was picked up with an inoculation loop and inoculated into 100 mL of MRS liquid medium and activated at 36°C for 20 h to obtain a Lactobacillus seed solution. The concentration of the Lactobacillus seed solution was approximately 2.1×10 7 cell / mL.

[0053] (2) Sterilize the ginseng seed oil separately in an autoclave at 115°C for 20 min.

[0054] (3) Sterilized ginseng seed oil and fermentation medium were mixed on a clean bench, and then 5% lactobacillus suspension was inoculated and incubated at 37°C and 150 r / min. -1 The cells were cultured under shaking conditions for 96 h.

[0055] (4) After the fermentation is completed, the mixture is placed in an autoclave at 80°C for 1 hour, and then the temperature is continued to be raised for high temperature sterilization (115°C). The mixture is then cooled, separated, and centrifuged to obtain the upper clear and transparent oily liquid to obtain the fermented ginseng seed oil.

[0056] The Lactobacillus gasseri CGMCC 1.3396 in step (1) was purchased from China General Microbiological Culture Collection Center.

[0057] The seed culture medium and fermentation medium in steps (1) and (3) are MRS culture medium: 10.0 g of peptone, 10.0 g of beef extract, 5.0 g of yeast powder, 20.0 g of glucose, 5.0 g of sodium acetate, 2.0 g of diamine hydrogen citrate, 1.0 mL of Tween-80, 2.0 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate heptahydrate, 0.05 g of manganese sulfate heptahydrate, and 1000 mL of distilled water. The pH is adjusted to 6.2-6.4 and sterilized in an autoclave at 115° C. for 15 min.

[0058] Comparative Example 2

[0059] The preparation method of the ginseng seed fermented oil of this comparative example comprises the following steps:

[0060] (1) Use an inoculation loop to pick Bacillus subtilis CGMCC A 1.361 and inoculate it into the seed culture medium. Activate it at 36°C for 10 h to obtain Bacillus subtilis seed solution. The concentration of the Bacillus subtilis seed solution is about 1.6×10 7 cell / mL.

[0061] (2) Sterilize the ginseng seed oil separately in an autoclave at 115°C for 20 min.

[0062] (3) Sterilized ginseng seed oil and fermentation medium were mixed on a clean bench, and then 5% Bacillus subtilis suspension was inoculated and incubated at 36°C and 200 r / min. -1 The cells were cultured under shaking conditions for 72 h.

[0063] (4) After the fermentation is completed, the mixture is placed in an autoclave at 80°C for 2 h, and then the temperature is continued to be raised for high temperature sterilization (115°C). The mixture is then cooled, separated, and centrifuged to obtain a clear and transparent oily liquid on the upper layer, thereby obtaining fermented ginseng seed oil.

[0064] Wherein, the Bacillus subtilis CGMCC A 1.361 in step (1) was purchased from China General Microbiological Culture Collection Center.

[0065] The seed culture medium in step (1) includes 3 g beef extract, 10 g peptone, 5 g NaCl, 1000 mL distilled water, pH 7.0, and high-pressure steam sterilization at 121° C. for 20 min.

[0066] The fermentation medium in step (3) includes: 4.0 g NH4NO3, 4.0 g K2HPO4, 6.0 g KH2PO4, 0.2 g MgSO4·7H2O, 1 mL trace element solution, 1.0 g yeast extract, 10.0 g glucose, pH 7.0, and 1000 mL deionized water. Sterilize at 115°C for 20 min.

[0067] The fermented ginseng seed oils were prepared in Comparative Examples 1 and 2, and the cream was prepared according to the method of Example 3, except that 4% of the ginseng seed oil fermented by lactobacillus and 4% of the ginseng seed oil fermented by Bacillus subtilis were used to replace 4% of the ginseng seed oil fermented by yeast.

[0068] Experimental Example 1

[0069] GC-MS analysis of ginseng seed oil before and after fermentation

[0070] Chromatographic conditions

[0071] Chromatographic column: HP-5MS capillary column (30m*250μm*0.25μm); stationary phase: 5% biphenyl-95% dimethylsiloxane, a non-polar capillary column; injection port temperature: 260°C, splitless injection; initial oven temperature: 50°C, maintained for 5 minutes, then increased to 250°C at 10°C / min, maintained for 5 minutes.

[0072] Mass spectrometry conditions: transfer line temperature: 280°C; acquisition mode: full scan; ion source temperature: 230°C; quadrupole: 160°C. The identification of the separated compounds was matched by computer spectral library NIST08.

[0073] Sample pretreatment method

[0074] Accurately weigh appropriate amounts of ginseng seed oil before and after fermentation in a conical flask, add n-hexane to dissolve, shake for 30 seconds, react at room temperature for 30 minutes, and take the supernatant for later use. The fermented ginseng seed oil is the yeast-fermented ginseng seed oil of Example 1.

[0075] The results of GC-MS analysis showed that the contents of 2,4-di-tert-butylphenol (2,4-DTBP), methyl palmitate (MEPA) and methyl stearate (MEST) in ginseng seed oil increased significantly after yeast fermentation, increasing by 1.23, 1.98 and 1.53 times respectively compared with those before fermentation. Figure 1 , 2 The contents of 2,4-DTBP, MEPA and MEST in ginseng seed oil before and after fermentation are shown in Tables 3 and 4.

[0076] Table 3 Names and contents of compounds in ginseng seed oil before fermentation

[0077]

[0078] Table 4 Names and contents of compounds in ginseng seed oil after fermentation

[0079]

[0080] Experimental Example 2

[0081] In vitro percutaneous absorption test

[0082] Preparation of ex vivo mouse skin: SD rats were anesthetized with an appropriate amount of isoflurane, and the hair on the abdominal skin was removed with an electric spatula. The rats were killed by cervical dislocation, and the abdominal skin was removed, subcutaneous fat and fascia were removed, and the rats were rinsed with saline, placed in saline, and stored in a refrigerator at 4°C. The skin was removed and checked for integrity on the second day of the experiment to avoid damage.

[0083] The treated rat skin stratum corneum was fixed upward on the Franz diffusion cell, 5 mL of 30% ethanol-normal saline was added to each receiving cell, and the cells were placed in an intelligent transdermal tester in a constant temperature water bath (the transdermal area was 0.5 cm 2 The experimental temperature was maintained at 35 ± 0.5 °C and the stirring speed was maintained at 350 ± 10 r·min -1 ). After 1 hour of equilibration, the air bubbles in the receiving pool were exhausted and then the drug was administered. 0.2g of glabridin cream was added to the supply pool, and each group was repeated 3 times. 1mL of sample was taken at 1, 2, 4, 6, 8, 10, 12, and 24 hours (the same amount of receiving solution at the same temperature was added in time), filtered through a 0.22μm microporous membrane, and analyzed. The cumulative permeation was calculated according to the following formula.

[0084] Calculation method of cumulative transdermal penetration: Calculate the concentration of glabridin and substitute it into the formula to calculate the cumulative penetration (Q n , μg / cm 2 ). The slope of the graph of cumulative permeation versus time (h) is the transdermal rate, i.e., the steady-state permeation rate (J s ). The permeability enhancement ratio (ER) is calculated by the formula: ER = J s / J s 0(J s 0: Steady-state permeation rate of the blank group; C n : concentration of the aspirate at the nth time point (mg / L); C i : The concentration of the aspirated solution at the nith time point (mg / L); V: The total volume of the receiving solution, i.e., 5 mL; V i : The volume of the receiving liquid absorbed each time is 1 mL; A: The area of ​​the receiving pool is 0.5 cm 2 ; C: drug concentration in the receiving pool)

[0085] Determination method of glabridin content

[0086] Chromatographic conditions: Column: Waters C 18 Column (100 mm × 2.1 mm, 1.7 μm); mobile phase: acetonitrile-water (60:40); detection wavelength: 280 nm; flow rate: 0.2 mL / min; injection volume: 2 μL; column temperature: 30°C; elution time: 8 min.

[0087] Preparation of reference solution: Take about 10.0 mg of glabridin reference, accurately weigh and place in a 10 mL brown volumetric flask, dissolve with methanol and make up to the mark to prepare a 1 mg / mL glabridin reference stock solution.

[0088] Skin sample treatment: The test skin was removed, rinsed with an appropriate amount of physiological saline, dried with filter paper, placed in a 1.5 mL centrifuge tube, cut into pieces with ophthalmic scissors, added with 1 mL of methanol, sealed and ultrasonically treated for 30 min at 10,000 r / min. -1 Centrifuge for 5 minutes, filter the supernatant with a 0.22 μm microporous filter membrane, and inject for analysis.

[0089] Standard curve drawing: dilute the glabridin reference stock solution to obtain a series of glabridin solutions of mass concentration, filter with a 0.22 μm microporous membrane, and inject for determination. Calculate the linear regression equation with glabridin concentration (X) as the horizontal axis and peak area (Y) as the vertical axis.

[0090] The transdermal penetration enhancing effects of glabridin creams containing unfermented ginseng seed oil, 4% yeast fermented ginseng seed oil (cream of Example 3), 4% Lactobacillus fermented ginseng seed oil (cream prepared in Comparative Example 1), and 4% Bacillus subtilis fermented ginseng seed oil (cream prepared in Comparative Example 2) were compared.

[0091] Table 5 Effects of different penetration enhancers on the penetration of glabridin cream

[0092]

[0093] Note: Compared with 4% ginseng seed oil group, *p<0.05.

[0094] As shown in Table 5, the results showed that ginseng seed oil before and after yeast fermentation could improve the transdermal absorption rate of glabridin, and the cumulative permeation amounts in 24 hours were 52.542, 31.625 and 33.186 μg / cm 2 , the penetration ratios were 2.95, 1.92 and 2.02 respectively. However, compared with the unfermented ginseng seed oil, the penetration enhancement effect of ginseng seed oil fermented with 4% yeast was significantly improved, while the penetration enhancement effect of ginseng seed oil fermented with Lactobacillus or Bacillus subtilis was not significantly improved.

[0095] Experimental Example 3

[0096] Effect of different amounts of yeast-fermented ginseng seed oil on the penetration enhancement of glabridin

[0097] The glabridin cream containing different concentrations of yeast-fermented ginseng seed oil prepared in Example 3 was tested for its transdermal penetration according to the method of Experimental Example 2.

[0098] Table 6 Regression equations and related parameters of different amounts of fermented ginseng seed oil glabridin cream

[0099]

[0100] Note: Compared with the blank group, *p<0.05.

[0101] Figure 3 As shown in Table 6, the penetration-enhancing effect of 3% fermented ginseng seed oil group>2% fermented ginseng seed oil group>1% fermented ginseng seed oil group>4% fermented ginseng seed oil group. When the concentration of fermented ginseng seed oil is less than or equal to 3%, the penetration-enhancing effect increases with the increase of concentration; when the concentration is 4%, the penetration-enhancing effect decreases, and there is no significant difference with 1%. This may be because MEPA and MEST and their analogs in ginseng seed oil have the effect of increasing the texture and consistency of the cream. Therefore, when the addition amount of fermented ginseng seed oil is less than or equal to 3%, the texture and consistency of the cream gradually increase, but it is not too thick, and does not affect the penetration-enhancing effect of fermented ginseng seed oil on glabridin; when the addition amount of fermented ginseng seed oil is greater than 3%, the texture of the cream is too thick, which is not conducive to the transdermal absorption of glabridin, which may also be the reason why the penetration-enhancing effect of 4% is the lowest.

[0102] Experimental Example 4

[0103] In order to further clarify the reasons for the improved penetration-enhancing effect of fermented ginseng seed oil, combined with the analysis results of GC-MS, the present invention further studied the penetration-enhancing effect of MEST, 2,4-DTBP and MEPA on glabridin, and prepared glabridin creams of corresponding concentrations according to Table 7.

[0104] Table 7 Recipe

[0105]

[0106] Table 8 Regression equations and related parameters of three kinds of glabridin cream

[0107]

[0108]

[0109] Note: Compared with the blank group, *p<0.05, **p<0.01.

[0110] Figure 4 The results in Table 8 show that the 24-hour cumulative permeation of glabridin by MEST, 2,4-DTBP and MEPA are 72.249, 62.076 and 119.032 μg / cm 2 The penetration enhancement ratios were 4.57, 3.76 and 7.47 respectively. Therefore, the reason why the penetration enhancement effect of fermented ginseng seed oil on glabridin is better than that of unfermented ginseng seed oil is related to the increase of the three components.

Claims

1. A composition for external use on skin comprising ginseng seed fermented oil having a penetration-enhancing effect, characterized in that: The skin topical composition is prepared from the following raw materials in the following weight proportions: 0.1-10 weight parts of a whitening active ingredient and 1-30 weight parts of ginseng seed fermented oil; the whitening active ingredient is selected from one or more of glabridin, arbutin, ellagic acid, niacinamide, kojic acid, ascorbyl glucoside, and ascorbyl tetraisopalmitate; the ginseng seed fermented oil is an oil obtained by fermenting ginseng seeds with yeast, and the ginseng seed fermented oil contains methyl palmitate.

2. The composition for external use on skin according to claim 1, characterized in that The ginseng seed fermented oil further comprises one or more of 2,4-di-tert-butylphenol and methyl stearate, and preferably further comprises 2,4-di-tert-butylphenol and methyl stearate.

3. The composition for external use on skin according to claim 1, characterized in that The whitening active ingredient is selected from glabridin or arbutin.

4. The composition for external use on skin according to claim 1, characterized in that The yeast is selected from one or more of baker's yeast, feed yeast, brewer's yeast, Pichia pastoris, and Candida.

5. The composition for external use on skin according to claim 1, characterized in that The ginseng seed fermented oil is prepared by the following method: after the ginseng seed oil is fermented by yeast, the fermented liquid is centrifuged to obtain the upper oily liquid.

6. A skin external preparation containing ginseng seed fermented oil having a penetration-enhancing effect, characterized in that: The preparation comprises the skin external composition according to any one of claims 1 to 5, and skin-acceptable auxiliary materials.

7. A skin cream for external use containing ginseng seed fermented oil with penetration-enhancing effect, characterized in that: Calculated by the mass of the cream, it includes 0.1%-1% of whitening active ingredients, 0.5-10% of ginseng seed fermented oil, 3%-8% of stearyl alcohol, 6%-15% of glyceryl monostearate, 1%-10% of vegetable oil or mineral oil or triglyceride of C4-10 fatty acid, 8%-15% of polyol, and the balance of water; the whitening active ingredient is selected from glycyrrhizin or arbutin, the ginseng seed fermented oil is an oil obtained by fermenting ginseng seeds with yeast, and the ginseng seed fermented oil contains methyl palmitate.

8. The skin external cream according to claim 7, characterized in that It includes 0.5-3% of ginseng seed fermented oil.

9. Use of the skin external composition according to any one of claims 1 to 5, the skin external preparation according to claim 6, or the skin external cream according to claim 7 or 8 in the preparation of a whitening external preparation.

10. Use of ginseng seed fermented oil in the preparation of an external composition for promoting skin penetration of whitening active ingredients, characterized in that: The ginseng seed fermented oil is oil obtained by fermenting ginseng seeds with yeast, and the ginseng seed fermented oil contains methyl palmitate.

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