Multi-effect skin-care liposome whitening composition and preparation method thereof

Abstract

The invention relates to the field of cosmetics, in particular to the field of A61K8, in particular to a multi-effect skin-care liposome whitening composition and a preparation method thereof. The multi-effect skin-care lipidosome whitening composition is prepared from the following raw materials: a whitening functional component, polyhydric alcohols, an emulsifier, a plant extract, lipidoids and deionized water, wherein the whitening functional component is prepared from glabridin, tranexamic acid, nicotinamide, acetylchitosamine, diglucosyl gallic acid, ectoin and ergothioneine. The multiple-effect skin-care liposome whitening composition has the effects of inhibiting the activity of tyrosinase, improving the DPPH clearance rate, inhibiting hyaluronidase and the like, and it is guaranteed that the product has excellent whitening and skin-care effects.

Description

Technical Field

[0001] The invention relates to the field of cosmetics, in particular to the field of A61K8, and specifically to a multi-effect skin care liposome whitening composition and a preparation method thereof. Background Art

[0002] Skin whitening has been a hot topic since ancient times. Whether it is the previous "white skin can cover up all ugliness", the enthusiasm and emphasis on whitening, or the current promotion of healthy skin color, even skin color, whitening and spot removal, the enthusiasm has been and will continue. Therefore, whitening products are still an important part of the cosmetics market, but on the road to whitening, traditional whitening ingredients may be irritating or the effect of using a single raw material is not obvious, and whitening cannot be inhibited from multiple channels. It is also the wrong use of whitening methods, blind use of instruments, or blind use of various active ingredients for stacking, which makes the whitening population not achieve a good whitening effect, but damages the skin barrier and makes the skin sensitive.

[0003] After the skin is stimulated by ultraviolet light, the signals of various pathways are transmitted, stimulating melanocytes to produce melanin, and transporting it from the basal layer to the stratum corneum. The production of melanin is regulated by "three enzymes and one factor", including tyrosinase (TYR), DHICA oxidase (TRP-1), dopachrome isomerase (TRP-2), and endothelin. These three enzymes are also regulated by MITF (microphthalmia transcription factor). At the same time, the signal pathway of endothelin also acts on MITF. Signal transduction stimulates the upregulation of MITF, thereby enhancing the activity of TYR, TRP-1, and TRP-2, and increasing melanin secretion. This signal transduction is not only caused by ultraviolet light, but can also be transmitted through inflammatory factors, free radicals, hormones in the body and other factors. Therefore, whitening at this stage should start from various pathways.

[0004] Traditional whitening pathways usually only inhibit the production of melanin by inhibiting the activity of tyrosinase or by inhibiting a single signal pathway. For example, the prior art CN112022776B discloses a whitening essence, the raw materials for its preparation include: scutellaria baicalensis root extract, glycyrrhiza glabra root extract, niacinamide, tranexamic acid, glycerin, xanthan gum, panthenol, and biosaccharide gum-1; it mainly achieves the whitening effect by inhibiting tyrosinase, and has relatively few inhibition channels.

[0005] As people's demand for whitening skin care increases, they not only emphasize streamlined skin care, advocate natural beauty, but also emphasize multifunctional skin care. However, the skin care raw materials in the existing technology often only have a single or limited function, which cannot meet consumers' demand for comprehensive skin care effects. Therefore, it is necessary to start from each channel, simplify the complex, select safe and targeted raw materials, correspond one by one, and break them down one by one to achieve whitening effects. Summary of the invention

[0006] In order to solve the above technical problems, the first aspect of the present invention provides a multi-effect skin care liposome whitening composition, the raw materials including: whitening functional ingredients, polyols, emulsifiers, plant extracts, lipids, deionized water; the whitening functional ingredients include: glycyrrhizin, tranexamic acid, niacinamide, acetyl glucosamine, diglucosyl gallic acid, ectoine and ergothioneine.

[0007] Preferably, the raw materials of the multi-effect skin care liposome whitening composition include, by mass percentage, 0.1%-1% whitening functional ingredient, 40%-60% polyol, 5%-20% emulsifier, 2%-10% plant extract, 2%-10% lipids, and the balance deionized water.

[0008] Whitening ingredients

[0009] Preferably, the mass ratio of glabridin, tranexamic acid, nicotinamide, acetyl glucosamine, diglucosyl gallic acid, ectoine and ergothioneine is (0.1-1): (0.1-2): (0.1-2): (0.1-2): (0.5-1): (0.5-1): (0.5-1).

[0010] The inventors found in actual research and development that the actual whitening effect of products with a single whitening pathway is relatively poor. When a specific whitening functional ingredient is selected for compounding, especially when glabridin, tranexamic acid, nicotinamide, acetyl glucosamine, diglucosyl gallic acid, ectoine and ergothioneine are selected for compounding, the transcription and / or protein expression of melanogenesis-related factors in cells can be significantly down-regulated synergistically, including melanocyte stimulating hormone receptor (MC1R), MITF, TYR, TRP-1 and TRP-2; and the whitening functional ingredient can effectively conduct the transmission of signals in the PKA / MITF and MAPK / MITF signaling pathways, effectively preventing black; in addition, the whitening functional ingredient can calm and soothe irritated and damaged skin, increase the regeneration process of the skin, inhibit the cascade effect caused by UVA, reduce the production of ICAM-1 induced by UVA radiation, and effectively reduce the content of DPPH in the cells, while blocking the stimulation of melanocytes from the source, inhibiting the conduction of signals in each pathway, and having multiple inhibitory effects on antioxidant sunscreen and anti-photoaging. However, the stability and permeability of the whitening ingredients in the whitening composition may be relatively poor.

[0011] Polyols

[0012] Preferably, the polyol includes: one or more of 1,2-propylene glycol, 1,3-propylene glycol, 1,3-butylene glycol, 1,2-pentanediol, 1,5-pentanediol, 1,2-hexanediol, glycerol, dipropylene glycol, sorbitol, caprylyl glycol, diethylene glycol methyl ether, diethylene glycol ethyl ether, and diethylene glycol butyl ether.

[0013] Further preferably, the polyol comprises glycerol and 1,2-hexanediol; the mass ratio of the two components is (40-60):(0.1-1).

[0014] Lipids

[0015] Preferably, the lipids include one or more of lecithin, hydrogenated lecithin, cholesterol, and phosphatidylcholine.

[0016] Further preferably, the lipid comprises hydrogenated lecithin.

[0017] As a carrier system, lipids can ensure the safety of whitening functional ingredients during storage and use. Its stable structure and good biocompatibility make the product safer and more reliable, reducing the safety risks caused by instability or leakage of whitening functional ingredients; in order to further improve the permeability and stability of whitening functional ingredients in the mask composition, the preferred hydrogenated lecithin in the present invention is lipids, and the introduction of hydrogenated lecithin greatly improves the stability of whitening functional ingredients. The increase in phase transition temperature and the saturation of molecular structure reduce the physical and chemical changes of whitening functional ingredients during storage and use, ensuring the long-term effectiveness of whitening composition products; at the same time, hydrogenated lecithin can combine with the skin to promote the skin's absorption of other ingredients in cosmetics, and the corresponding whitening functional ingredients are more likely to penetrate into the deep layers of the skin, thereby improving the absorption effect and achieving better skin care effects.

[0018] Emulsifier

[0019] Preferably, the emulsifier includes: one or more of polyoxyethylene sorbitan laurate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monooleate, sorbitan stearate, sorbitan sesquioleate, sorbitan trioleate, sorbitan stearate, sorbitan isostearate, caprylic / capric triglyceride, polyglyceryl-2 diisostearate, and polyglyceryl-6 polyricinoleate.

[0020] Further preferably, the emulsifier is caprylic / capric triglyceride.

[0021] In the present invention, caprylic acid / capric acid triglyceride is preferably used as an emulsifier, which can reduce the interaction force between the lipid particles encapsulating the whitening functional ingredients, prevent their aggregation and deposition, thereby maintaining the stability of the lipid structure, and making the whitening composition have better skin care and whitening effects.

[0022] Plant Extracts

[0023] Preferably, the plant extract comprises: one or more of shea butter (Butyrospermum Parkii), camellia seed oil (Camellia Japonica), olive oil (Olea Europaea), peony seed oil (Paeonia Suffruticosa Andr.), sweet almond oil (Prunusdulcis Var.Dulcis), jojoba oil (Simmondsia Chinensis), cactus seed oil (Opuntia Ficus-Indica), grape seed oil (Vitis Vinifera), and rosehip oil (Rosa Damascena).

[0024] Further preferably, the plant extract is olive oil.

[0025] Olive oil is rich in unsaturated fatty acids and natural moisturizing factors, which can effectively lock in moisture, prevent skin moisture loss, and keep the skin moisturized and smooth. In liposome cosmetics, olive oil can be used as the main moisturizing ingredient to provide long-lasting moisturizing effects for the skin; at the same time, liposomes, as an efficient carrier system, can better transport the moisturizing ingredients in olive oil to the deep layers of the skin, improving the moisturizing effect.

[0026] The second aspect of the present invention provides a method for preparing a multi-effect skin care liposome whitening composition, comprising:

[0027] S1, mixing polyols, lipids and deionized water, and heating to completely dissolve the raw materials to form phase A;

[0028] S2, mixing the emulsifier, the plant extract, and the whitening functional ingredient to form phase B;

[0029] S3. Add phase B to phase A for homogenization, stirring and emulsification. After the emulsification is completed, cool the liquid to room temperature and put it into a high-pressure homogenizer. Cycle high-pressure homogenization and emulsification for 4-5 times until the material is clear and translucent and then discharge it.

[0030] Preferably, the heating temperature in step S1 is 70-85°C.

[0031] Preferably, in the step S3, the stirring and homogenizing speed is controlled at 1800-2500 rpm, and the homogenizing pressure of the high-pressure homogenizer is 850-900 bar.

[0032] High-pressure homogenizer can significantly improve the stability and encapsulation rate of liposomes when preparing liposome cosmetics. High-pressure homogenizer optimizes the rotation speed and pressure during the homogenization process, and passes the liposome solution through a narrow slit in a very short time, so that the solution is subjected to strong shear, impact and cavitation effects, thereby achieving the purpose of homogenization, making the liposome more compact and stable, reducing the leakage and degradation of whitening functional ingredients, thereby improving the overall stability of the whitening composition product and the whitening skin care effect.

[0033] Beneficial Effects

[0034] (a) The present invention uses specific whitening ingredients, including glycyrrhizin, tranexamic acid, niacinamide, acetyl glucosamine, diglucosyl gallic acid, ectoine and ergothioneine, to ensure that the product has excellent whitening and skin care effects from the aspects of inhibiting tyrosinase activity, improving DPPH clearance rate, inhibiting hyaluronidase, etc.

[0035] (ii) In addition to whitening ingredients, the present invention selects a variety of whitening raw material combinations, starting from several stages of melanin production, which can not only inhibit ultraviolet damage to cells from the source, reduce the production of free radicals and inflammatory factors, and block the stimulation of melanocytes from the source, but also inhibit the conduction of signals in various pathways, reduce / lower the number / activity of three enzymes related to melanin production, and inhibit the production of melanin. Finally, inhibit the transport of melanin to the stratum corneum, accelerate the metabolism of melanin in the stratum corneum, and block melanin layer by layer. Under the premise of not harming the cells themselves and damaging the body, it can effectively and healthily block melanin.

[0036] (III) The raw materials for preparing the components of the present invention are healthy and highly safe, and are suitable for use on all skin types.

[0037] (iv) The present invention adopts liposome embedding technology, which can ensure that the inactivation rate of whitening functional ingredients is reduced while increasing the transdermal absorption rate of raw materials and reducing the irritation of some raw materials to sensitive skin.

[0038] (V) The preparation method of the multi-effect skin care liposome whitening composition provided by the present invention is simple, and the product can be produced on a large scale industrially. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is a trend chart of the tyrosinase inhibition rate of the product in Example 1.

[0040] Figure 2 This is a trend chart of the DPPH clearance rate of the product in Example 1.

[0041] Figure 3 This is a diagram showing the hyaluronic acid inhibition rate effect of the product in Example 1. DETAILED DESCRIPTION

[0042] Example 1

[0043] The first aspect of this example provides a multi-effect skin care liposome whitening composition, the raw materials, calculated by mass percentage, include: 1% whitening functional ingredient, 50% polyol, 5% emulsifier, 5% plant extract, 5% lipid, and the balance deionized water.

[0044] The whitening ingredients are: glabridin, tranexamic acid, nicotinamide, acetyl glucosamine, diglucosyl gallic acid, ectoine and ergothioneine; the mass ratio is 1:2:2:2:1:1:1.

[0045] The glabridin is from Luoyang Lansley Technology Co., Ltd.; the trade name of tranexamic acid is BINACTI CM, which is from Shanghai Zibang Biopharmaceutical Co., Ltd.; the trade name of nicotinamide is BINACTI CM, which is from Shandong Kunda Biotechnology Co., Ltd.; the trade name of acetyl glucosamine is: Acetyl glucosamine, sourced from Shandong Tiansheng Biotechnology Co., Ltd.; diglucosyl gallic acid, trade name: BRIGHTENYL, sourced from Givaudan Daily Flavors and Fragrances (Shanghai) Co., Ltd.; tetrahydromethylpyrimidine carboxylic acid (Ectoine), trade name: Ectoine, sourced from Shanghai Yinong Cosmetics Technology Center; ergothioneine, sourced from Shanghai Yinong Cosmetics Technology Center.

[0046] The polyol includes glycerol and 1,2-hexanediol, and the mass ratio of the two components is 49:1; the trade name of glycerol is APICERINE G997 USP, which is derived from PT SARIDUMAI SEJATT; the trade name of 1,2-hexanediol is: 60, from Dezhixin Flavors and Fragrances (Nantong) Co., Ltd.

[0047] The emulsifier is caprylic / capric triglyceride, trade name: 318RC, from BASF (China) Co., Ltd.

[0048] The plant extract is olive oil, purchased from Northstar Lipids (UK) Ltd, trade name: Olive Oil CB olive oil.

[0049] The lipid is hydrogenated lecithin, trade name: Soya SPL 75H hydrogenated lecithin, sourced from Suzhou Nacon Biotechnology Co., Ltd.

[0050] The second aspect of this example provides a method for preparing a multi-effect skin care liposome whitening composition, comprising:

[0051] S1, mixing polyol, lipid and deionized water, heating to 80°C, completely dissolving the raw materials to form phase A;

[0052] S2, mixing the emulsifier, the plant extract, and the whitening functional ingredient to form phase B;

[0053] S3. Keep the temperature of phase A at 80°C, then add phase B, and stir homogenously at 2000rpm until emulsification is complete; cool the liquid after complete emulsification to room temperature 25°C, put it into a high-pressure homogenizer, set the pressure to 900bar, and cycle high-pressure homogenization and emulsification at 2000rpm for 5 times until the material is clear and translucent, then discharge the material.

[0054] Comparative Example 1

[0055] This example provides a multi-effect skin care liposome whitening composition. The specific implementation method is the same as Example 1, except that the whitening functional ingredient does not include glabridin, but is replaced by an equal mass portion of 377 (phenethyl resorcinol).

[0056] However, preliminary tests of this product showed that the stability and whitening effect (tyrosinase activity inhibition effect and DPPH test rate) were not as good as those in Example 1, and the practicability was not strong, so no subsequent efficacy verification test was performed.

[0057] Performance Testing

[0058] 1. Whitening efficacy test (inhibition of tyrosinase activity)

[0059] Principle: Tyrosinase is a key enzyme in the biosynthesis of skin melanin. It acts on DOPA to form DOPA quinone, which spontaneously undergoes a series of reactions to finally form melanin. In a phosphoric acid solution with a pH of 6.8, tyrosinase can catalyze the conversion of DOPA into DOPA quinone, and the absorbance can be measured at 475nm on a spectrophotometer. Raw materials with tyrosinase activity inhibition can reduce the conversion of DOPA into DOPA aldehyde, thereby reducing the absorbance value. According to the change in absorbance value, the inhibitory effect of the raw material on tyrosinase activity is evaluated. This test analyzes the whitening ability of the test object by detecting the change in absorbance. The experimental method is implemented in accordance with the group standard T / SHRH015-2018 "Cosmetics-Tyrosinase Activity Inhibition Experimental Method".

[0060] 1.1 Main Reagents

[0061] Tyrosinase, L-dopa, disodium hydrogen phosphate dodecahydrate, citric acid monohydrate, ethanol.

[0062] 1.2 Main instruments

[0063] Microplate reader (TECAN, Infnite F50), analytical balance, adjustable pipette, 96-well plate 1.3 Positive control

[0064] In this experiment, kojic acid was used as the positive control and was dissolved and diluted with the prepared pH 6.8 buffer into five concentration gradients of 1 mg / mL, 0.5 mg / mL, 0.25 mg / mL, 0.125 mg / mL and 0.0625 mg / ml to verify the test system.

[0065] 2. Experimental Procedure

[0066] 2.1 Solution preparation

[0067] 2.1.1 Sample information and test concentration

[0068] The concentration information of the test samples and control samples is shown in Table 1

[0069] Table 1

[0070]

[0071] 2.1.2 Tyrosinase solution extraction

[0072] Peel and chop the potatoes, weigh 100g, and freeze them at -20℃ overnight. Take the frozen potatoes, add 100mL of disodium hydrogen phosphate-citric acid buffer with pH=6.8, and use a juicer to make a homogenate. Pour the potato homogenate into a centrifuge tube, centrifuge at 4000r / min, centrifuge at 4℃ for 15min, take out the supernatant, aspirate it, dilute it 2 times, store it at 4℃, and use it up within 2h.

[0073] 2.2 Sample addition

[0074] The experiment set up enzyme reaction wells, solvent background wells, sample reaction wells and sample background wells. Refer to Table 2, add each solution in order, incubate at 37°C for 10 minutes, and finally add L-DOPA solution to react for 5 minutes, and then measure the absorbance value near 475nm.

[0075] Table 2

[0076]

[0077]

[0078] 2.3 Determination of tyrosinase activity inhibition rate

[0079] Tyrosinase inhibition rate (%) = 1-[(T-T0) / (C-C0)] × 100%

[0080] 3. Experimental results

[0081] According to the specific experimental method, the tyrosinase inhibition rate was calculated. The results are shown in Figure 3, and the change trend is shown in Figure 1. The details are as follows:

[0082] Table 3

[0083]

[0084] Note: The tyrosinase inhibition rate of the sample at each tested concentration needs to be calculated based on the standard deviation of the inhibition rate between parallel tubes in each group. The SD value must be ≤3%, and the parallelism of the test is considered valid.

[0085] The test results show that the whitening composition of Example 1 has excellent tyrosinase inhibition, and the tyrosinase inhibition rate at the same concentration gradient is higher than that of kojic acid, with the highest inhibition rate being 93.9%. And the tyrosinase inhibition rate is still relatively high at a concentration of 0.0625%.

[0086] (Note: The inhibition rate first increased and then decreased for the following reasons: In the experiment, different concentrations of raw material combinations were added to tyrosinase at the same time. When the concentration of the raw material combination was 1%, the state was relatively viscous, and the reaction rate was affected to a certain extent).

[0087] 2. Antioxidant test of whitening composition (DPPH scavenging rate)

[0088] Principle: 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable long-lived free radical. Its ethanol solution is dark purple and has strong absorption near 517nm. When there is a free radical scavenger, the DPPH free radical is partially scavenged and the number is reduced. The absorbance A value at the maximum absorption wavelength will decrease and the color will become lighter. The fading degree of the DPPH ethanol solution is linearly related to the number of electrons it accepts. This can be used to evaluate the ability of the test sample to scavenge free radicals, that is, the size of the antioxidant activity.

[0089] This test analyzes the antioxidant capacity of the test object by detecting the changes in the DPPH free radical content. The experimental method is implemented with reference to the group standard T / SHRH006-2018 "Cosmetics-Free Radical (DPPH) Scavenging Experiment Method".

[0090] 4.1 Main Reagents

[0091] DPPH, vitamin E, 95% volume concentration ethanol, water.

[0092] 4.2 Main instruments

[0093] Microplate reader (TECAN, Infnite F50), analytical balance, adjustable pipette, 96-well plate 4.3 Positive control

[0094] In this experiment, vitamin E was used as the positive control and was diluted with 95% volume concentration ethanol to five concentration gradients of 0.08 mg / mL, 0.04 mg / mL, 0.02 mg / mL, 0.01 mg / mL and 0.005 mg / mL to verify the test system.

[0095] 5. Experimental Procedure

[0096] 5.1 Solution preparation

[0097] 5.1.1 Sample information and test concentration

[0098] The concentration information of the test samples and control samples is detailed in Table 4.

[0099] Table 4

[0100]

[0101] 5.1.2 Pre-prepare 0.12 mg / mL DPPH ethanol solution:

[0102] Weigh 12 mg of 1,1-diphenyl-2-trinitrophenylhydrazine into a beaker, add 10 mL of 95% ethanol, stir until dissolved, transfer to a centrifuge tube, and mix with a mixer for 5 minutes to ensure that the solution is uniform from top to bottom. DPPH solution is unstable and should be prepared and used immediately.

[0103] 5.2 Sample addition

[0104] 5.2.1 Repetition setting

[0105] Referring to Table 5, set up sample wells (T), sample background (T0), DPPH wells (C) and solvent background (C0). For each sample well (T) of each tested concentration, three replicate wells need to be set up, and three replicate wells also need to be set up for the DPPH well (C).

[0106] 5.2.2 Add test sample

[0107] Add 50 mL of sample solution of the same concentration to the sample well (T) and sample background (T).

[0108] 5.2.3 Replenishing solvent

[0109] Solvents were added to all test tubes (T, T0, C, C0). For water-soluble samples, water was used; for oil-soluble samples, 95% volume concentration of ethanol was used. The volume was made up to 150 μL and mixed.

[0110] 5.2.4 Addition of DPPH

[0111] Add 50 μL of DPPH ethanol solution to the sample well (T) and DPPH well (C), replace the sample background (T) and solvent background (C0) with 95% ethanol, shake gently, and let stand at room temperature for 5 minutes.

[0112] Table 5

[0113]

[0114] 5.3 DPPH content determination

[0115] The absorbance was measured at around 517 nm using an enzyme reader.

[0116] 5.4 DPPH clearance calculation

[0117] DPPH clearance rate (%) = 1-[(T-T0) / (C-C0)]×100%

[0118] 6. Experimental Results

[0119] According to the specific experimental method, the DPPH free radical scavenging rate of the whitening composition of Example 1 was calculated. The results are shown in Table 6. The change trend is as follows: Figure 2 As shown, the details are as follows:

[0120] Table 6

[0121]

[0122]

[0123] Note: IC of positive control vitamin E 50 The test system is considered valid if the sample concentration (the concentration corresponding to the 50% scavenging effect) is between 0.02mg / mL and 0.06mg / mL. The free radical scavenging rate of the sample at each tested concentration is calculated, and the standard deviation of the scavenging rate between parallel tubes in each group is calculated. The SD value must be <3%, and the parallelism of the test is considered valid.

[0124] The test results showed that the IC 50 Within the range specified by the standard, the experimental system is verified to be effective; the whitening raw material combination sample has a certain free radical scavenging ability for DPPH free radicals at the tested concentration, which can be used as one of the evidences for its antioxidant effect; in order to eliminate the influence of solution turbidity on the data as much as possible, the DPPH scavenging rate of the sample at a low concentration of 0.313% is selected. The sample in Example 1 still has a good scavenging rate for DPPH free radicals at low concentrations, and it is believed that the sample has good antioxidant properties.

[0125] 3. Whitening combination soothing test (hyaluronidase inhibition)

[0126] Principle: According to the immune response mechanism, allergic reactions are divided into type I, type II, type Ш and type IV. Most allergic reactions are type I allergic reactions related to the activity of hyaluronidase in the body. The degradation products of hyaluronic acid can increase the inflammatory response. The degree of reaction depends on the activity of hyaluronidase. Therefore, the ability of a substance to inhibit the activity of hyaluronidase can be used as an indicator of its anti-inflammatory response. In an acidic environment, hyaluronidase can hydrolyze the substrate and convert it into N-acetylglucosamine, which produces a color reaction with dimethylaminobenzaldehyde. The experiment evaluates the ability of the sample to inhibit the activity of hyaluronidase by comparing the changes in absorbance before and after the reaction, thereby judging its soothing effect.

[0127] 7. Materials

[0128] 7.1 Main Reagents

[0129] Glacial acetic acid; anhydrous sodium acetate; hyaluronidase (biotechnology grade, 300μ / mg-500μ / mg); sodium hyaluronate (from Streptococcus equi); sodium carbonate; acetylacetone; p-dimethylaminobenzaldehyde (CAS No. 100-10-7); concentrated hydrochloric acid (37%); anhydrous ethanol; anhydrous calcium chloride; sodium oxide; dipotassium glycyrrhizinate (purity ≥98%).

[0130] 7.2 Main instruments

[0131] Microplate reader, analytical balance, adjustable pipette, 96-well plate, test tubes, water bath.

[0132] 7.3 Positive Control

[0133] In this experiment, 2 mg / mL of dipotassium glycyrrhizinate was used to verify the test system.

[0134] 8. Experimental Procedure

[0135] 8.1 Solution preparation

[0136] The mass concentration of the whitening composition of Example 1 used in this experiment is 1%.

[0137] 8.2 Sample addition

[0138] 8.2.1 Test groups

[0139] The experiment set up sample groups, sample background groups, enzyme reaction groups and solvent background groups. Three replicate wells were required for each sample group of the tested concentration of the raw material, and three replicate wells were also required for the enzyme reaction group.

[0140] 8.2.2 Test steps

[0141] Refer to Table 7, add reagents in the test tube in order, and after the reaction is completed, read at 530nm.

[0142] Table 7

[0143]

[0144]

[0145] 8.3 Calculation of hyaluronidase inhibition rate

[0146]

[0147] 9. Experimental Results

[0148] According to the specific experimental method, the hyaluronic acid inhibition rate was calculated. The results are shown in Table 8. The change trend is as follows Figure 3 As shown, the details are as follows.

[0149] Table 8

[0150]

[0151] Note: Statistical analysis was performed using t-test. When the PC group (positive control group) and the sample group were compared with the NC group (negative control group), a P value <0.05 indicated a significant difference, indicated by "*", and a P value <0.01 indicated an extremely significant difference, indicated by "**".

[0152] Statistical analysis showed that the PC group was P-0.001, less than 0.05, compared with the NC group, and the inhibition rate of the PC group was 13.39%, which was in line with the standard range, verifying the effectiveness of the experimental system; compared with the NC group, the P value of the whitening composition of Sample Example 1 was 0.000, less than 0.01, showing a very significant difference.

[0153] According to the test results, the hyaluronidase inhibition rate of the PC group (2 mg / mL dipotassium glycyrrhizinate) in this test was 13.39%, which was in line with the standard range, verifying the effectiveness of the experimental system; at the test concentration, the average hyaluronidase inhibition rate of the whitening composition of Sample Example 1 was 87.98% SD<15% and compared with the NC group, the P value was <0.05, which means that the sample has a certain soothing effect.

Claims

1. A multi-effect skin care liposome whitening composition, characterized in that: Raw materials include: whitening ingredients, polyols, emulsifiers, plant extracts, lipids, and deionized water; The whitening ingredients include: glabridin, tranexamic acid, nicotinamide, acetyl glucosamine, diglucosyl gallic acid, ectoine and ergothioneine.

2. The multi-effect skin care liposome whitening composition according to claim 1, characterized in that: The raw materials of the multi-effect skin care liposome whitening composition include, by mass percentage, 0.1%-1% whitening functional ingredient, 40%-60% polyol, 5%-20% emulsifier, 2%-10% plant extract, 2%-10% lipid, and the balance deionized water.

3. The multi-effect skin care liposome whitening composition according to claim 1, characterized in that: The mass ratio of glabridin, tranexamic acid, nicotinamide, acetyl glucosamine, diglucosyl gallic acid, ectoine and ergothioneine is (0.1-1): (0.1-2): (0.1-2): (0.1-2): (0.5-1): (0.5-1): (0.5-1).

4. The multi-effect skin care liposome whitening composition according to claim 1, characterized in that: The polyols include: one or more of 1,2-propylene glycol, 1,3-propylene glycol, 1,3-butylene glycol, 1,2-pentanediol, 1,5-pentanediol, 1,2-hexanediol, glycerol, dipropylene glycol, sorbitol, caprylyl glycol, diethylene glycol methyl ether, diethylene glycol ethyl ether and diethylene glycol butyl ether.

5. The multi-effect skin care liposome whitening composition according to claim 1, characterized in that: The lipids include one or more of lecithin, hydrogenated lecithin, cholesterol and phosphatidylcholine.

6. The multi-effect skin care liposome whitening composition according to claim 1, characterized in that: The plant extract comprises one or more of shea butter, camellia seed oil, olive oil, peony seed oil, sweet almond oil, jojoba oil, cactus seed oil, grape seed oil and rose hip oil.

7. The multi-effect skin care liposome whitening composition according to claim 1, characterized in that: The emulsifier includes: one or more of polyoxyethylene sorbitan laurate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monooleate, sorbitan stearate, sorbitan sesquioleate, sorbitan trioleate, sorbitan stearate, sorbitan isostearate, caprylic / capric triglyceride, polyglyceryl-2 diisostearate, and polyglyceryl-6 polyricinoleate.

8. A method for preparing the multi-effect skin care liposome whitening composition according to any one of claims 1 to 7, characterized in that: The following steps are involved: S1, mixing polyol, lipid and deionized water, heating until completely dissolved to form phase A; S2, mixing the emulsifier, the plant extract, and the whitening functional ingredient to form phase B; S3. Add phase B to phase A for homogenization, stirring and emulsification. After the emulsification is completed, cool the liquid to room temperature and put it into a high-pressure homogenizer. Cycle high-pressure homogenization and emulsification for 4-5 times until the material is clear and translucent and then discharge it.

9. The method for preparing the multi-effect skin care liposome whitening composition according to claim 8, characterized in that: The heating temperature in the step S1 is 70-85°C.

10. The method for preparing the multi-effect skin care liposome whitening composition according to claim 8, characterized in that: In the step S3, the stirring and homogenizing speed is controlled at 1800-2500 rpm, and the homogenizing pressure of the high-pressure homogenizer is 850-900 bar.

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