Plant extract composition with whitening effect and preparation method thereof

Abstract

The invention belongs to the technical field, and particularly relates to a plant extract composition with a whitening effect and a preparation method thereof. The composition provided by the invention takes natural plants and bioactive components as cores, avoids the use of chemical whitening agents with potential toxicity, such as hydroquinone, mercury salt and the like, and is high in safety and low in irritation. All the components cooperate with one another in a reasonable proportion, so that melanin generation can be inhibited from the source, and metabolism of generated melanin can be promoted; according to the present invention, the whitening effect is improved, the multiple skin care functions such as moisturizing, anti-aging, repairing and the like are further considered, and the mask is suitable for various skin types, and is especially suitable for people with dark skin color, hyperpigmentation, dryness, water shortage and mild sensitive skin.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field, and particularly relates to a plant extract composition with whitening effect and a preparation method thereof. BACKGROUND

[0002] With the improvement of people's living standards and the increasing attention to skin health, whitening skin care products have become an important category in the global cosmetics market. Skin pigmentation is mainly caused by uneven generation and distribution of melanin, and the synthesis of melanin mainly occurs in melanocytes in the basal layer of the skin epidermis, and the key enzyme is tyrosinase. The enzyme catalyzes the hydroxylation of tyrosine to DOPA, and further oxidizes to dopaquinone, and finally forms melanin through a series of reactions. Therefore, inhibiting the activity of tyrosinase is considered to be one of the effective ways to achieve skin whitening.

[0003] At present, the common whitening active ingredients on the market mainly include chemical synthesis or semi-synthesis substances such as hydroquinone, arbutin, vitamin C and its derivatives, kojic acid, azelaic acid. Although these ingredients have a whitening effect to some extent, long-term use may be accompanied by skin irritation, allergy, pigment rebound, even toxicity and other safety problems. For example, although hydroquinone has strong whitening effect, it has been restricted in many countries due to its potential cytotoxicity and carcinogenic risk. Therefore, the development of safe, efficient and natural whitening ingredients has become an important direction of current cosmetic research and development.

[0004] Plant extracts have attracted widespread attention due to their wide sources, diverse biological activities and high safety. A large number of studies have shown that many plants contain active ingredients that can inhibit the activity of tyrosinase, scavenge free radicals, antioxidant or block the transmission of melanin, such as flavonoids, polyphenols, saponins and alkaloids. However, single plant extract often has limited whitening effect, poor stability, low transdermal absorption rate and other problems, which is difficult to meet the demand of efficient whitening products. In addition, different plant ingredients may have antagonistic effects, affecting the overall efficacy. Therefore, it has important application value and market prospect to develop a composition based on natural plants, which has high efficiency, good stability and high safety by reasonably matching multiple effective extracts. SUMMARY

[0005] The present application aims to provide a plant extract composition with whitening effect and a preparation method thereof, in order to overcome the defects in the prior art and meet the urgent needs of consumers for safe, green and efficient whitening products.

[0006] The present application provides a kind of plant extract composition with whitening effect, the composition includes python oil 1~5 parts, safflower extract 0.5~3 parts, osmanthus extract 0.1~2 parts, pearl hydrolysate 0.5~2 parts, oyster extract 0.3~1.5 parts, madder extract 0.3~1.5 parts, radix lithospermi extract 0.2~1 part, fish collagen peptide 1~4 parts, glycerol 5~12 parts, silk protein 0.2~1 part, ergothioneine 0.02~0.2 part, vitamin C 0.5~3 parts and vitamin E 0.1~1 part by mass parts.

[0007] Preferably, the content of hydroxyl safflower yellow G in the safflower extract is 5%wt~10%wt;The content of verbascoside in the osmanthus extract is 15%wt~30%wt;The content of alizarin in the madder extract is ≥5%wt;The content of lithospermum in the radix lithospermi extract is 2%wt~5%wt.

[0008] Preferably, the average molecular weight of the fish collagen peptide is ≤1000Da, and the hydroxyproline content is ≥8%wt;The content of hydrolyzed silk protein in the silk protein is ≥90%wt, and the molecular weight of the silk protein is 500~3000Da.

[0009] The present application also provides a kind of liposome with whitening effect, the liposome includes the composition described in the above technical solution.

[0010] The present application also provides a preparation method of the liposome described in the above technical solution, the method includes the following steps: The film-forming lipid, python oil, safflower extract, osmanthus extract, madder extract, radix lithospermi extract and vitamin E are dissolved in anhydrous ethanol to obtain phase A; The pearl hydrolysate, oyster extract, fish collagen peptide, glycerol, silk protein, ergothioneine and vitamin C are dissolved in PBS solution to obtain phase B;Phase A is injected into the organic phase inlet of the microfluidic chip at a flow rate of 0.4~0.6 mL·min -1 Supercritical CO2 is injected into the anti-solvent inlet of the microfluidic chip at a flow rate of 2.5~3.5 mL·min -1 The reaction is carried out at 25℃~30℃ and 7~15 MPa to form porous lipid microparticles, which are collected as dry powder at the chip outlet after 2~5 s of residence time;Phase B is added to the porous lipid microparticle dry powder and shaken to form liposomes;The film-forming lipid is a mixture of hydrogenated lecithin, cholesterol and distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000 (DSPE-PEG2000) in a mass ratio of 5.5~6.5:2.8~3.2:0.9~1.1; Preferably, 5mL of the A phase and 5mL of the B phase are prepared per g of the composition according to the technical solution, and 1-2g of the film-forming lipid is used per g of the composition according to the technical solution.

[0011] Preferably, the porous lipid microparticles have a particle size of <100nm.

[0012] Preferably, the temperature of the B phase is 2-8℃.

[0013] Preferably, the shaking condition is 40-80rpm oscillation for 1-3min.

[0014] The present application also provides a whitening cream containing the liposome according to the technical solution.

[0015] The present application has the following advantages: The composition provided by the present application is rich in various natural whitening active ingredients, wherein the silk amino acids in the vitamin C and the silk fibroin can effectively inhibit the activity of tyrosinase, block the formation of dopaquinone, and reduce the synthesis of melanin; the extracts of madder and comfrey have good antioxidant and anti-inflammatory effects, can regulate the activity of melanocytes, and reduce pigmentation; the pearl hydrolysate is rich in various amino acids and trace elements, and can brighten the skin color and improve sallowness; the osmanthus extract can synergize with other ingredients, and thus plays a role in uniform skin color. The composition of the present application can inhibit the generation of melanin from the source and promote the metabolism of the generated melanin through the synergistic effect of various ingredients. The fish collagen peptide can accelerate the renewal of keratinocytes, promote the shedding of the generated melanin with the keratin, remove free radicals, reduce the deepening of the pigmented spots induced by oxidative stress, enhance the integrity of the stratum corneum, and reduce inflammation caused by external stimulation.

[0016] Vitamin E and vitamin C can synergistically exert antioxidant effect, prevent lipid peroxidation, and protect the integrity of the cell membrane; the safflower extract is rich in safflower yellow pigment, and has significant free radical scavenging and anti-photoaging ability. The composition can effectively resist oxidative stress caused by ultraviolet rays and environmental pollution, and prevent the formation of pigmented spots and skin aging.

[0017] Glycerol has strong hygroscopicity and can absorb water from the environment; silk fibroin is rich in hydrophilic amino acids and can improve the water content of the stratum corneum; fish collagen peptide has a small molecular weight and is easy to penetrate, and can supplement the lost collagen of the skin and enhance the elasticity and tightness of the skin. The three components synergistically act together to significantly improve the water retention capacity of the skin, improve dryness and roughness, and make the skin soft and lustrous, thereby forming an efficient moisturizing network.

[0018] Snake oil is rich in unsaturated fatty acids and ceramides, with excellent permeability and biocompatibility, which can nourish the deep layers of the skin and repair the damaged barrier; oyster extract is rich in trace elements such as zinc and selenium, which helps regulate skin metabolism and promote cell regeneration; comfrey and madder have anti-inflammatory and soothing effects, which can reduce skin irritation and are suitable for gentle whitening care for sensitive skin.

[0019] Ergothioneine and vitamin E can provide synergistic protection for other active ingredients, significantly extending product shelf life and showing promising prospects for industrial application.

[0020] The composition provided by this invention uses natural plants and bioactive ingredients as its core, avoiding the use of potentially toxic chemical whitening agents such as hydroquinone and mercury salts. It is highly safe and has low irritation. The components work synergistically in a reasonable ratio, not only enhancing whitening efficacy but also providing multiple skincare functions such as moisturizing, anti-aging, and repair. It is suitable for various skin types, especially for those with dull skin, pigmentation, dryness, and mild sensitivity.

[0021] The composition provided by this invention has good physicochemical stability, is not prone to separation, discoloration or inactivation, and can be widely used in various cosmetic formulations such as emulsions, serums, creams, and masks, and has good industrialization prospects and market application value.

[0022] This invention employs a one-step "microfluidic-supercritical CO2 instantaneous anti-dissolution-recrystallization" method. Under mild conditions of low temperature, chloroform-free, and low shear, it precisely encapsulates multiple lipid-soluble and water-soluble active ingredients in distinct zones. Lipid-soluble components are embedded in the lipid bilayer, while water-soluble components are locked within the internal aqueous phase, achieving an overall encapsulation rate of 85%–95%, significantly superior to traditional thin-film-hydration methods. The preparation process of this invention involves no high temperatures, no ultrasound, and no organic solvent residues, avoiding degradation of heat-sensitive or easily oxidized components such as peptides and vitamin C. Experimental verification shows that the liposomes prepared by this invention retain >90% activity after 30 days of storage at 4°C, exhibiting excellent stability. The microfluidic chip combined with the instantaneous anti-dissolution process allows for control of the liposome particle size to 80–120 nm, with a PDI <0.15. The system is transparent and stable, facilitating transdermal absorption and improving skin feel. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0024] Figure 1 This is a schematic diagram of a face cream prepared from liposomes according to Example 1 of the present invention. Detailed Implementation

[0025] In order to further illustrate the present application, the solutions provided by the present application are described in detail below in conjunction with examples, but they should not be understood as limiting the protection scope of the present application.

[0026] Unless otherwise specified, the substances used in the embodiments of the present application are all conventional commercially available products.

[0027] The present application does not have special requirements for the sources of each extract, and all conventional commercially available products can be used. In the embodiments of the present application, the safflower extract is purchased from Snowlot Biological Technology Co., Ltd., wherein the content of hydroxysafflor yellow A is 8%wt; the Osmanthus fragrans extract is purchased from Snowlot Biological Technology Co., Ltd., wherein the content of verbascoside is 20%wt; the madder extract is purchased from Snowlot Biological Technology Co., Ltd., wherein the content of alizarin is 6%wt; the radix lithospermi extract is purchased from Snowlot Biological Technology Co., Ltd., wherein the content of shikonin is 5%wt; the pearl hydrolysate is purchased from Guangzhou Anpin Chemical Co., Ltd.; and the oyster extract is purchased from Shanxi Xintianyu Biological Technology Co., Ltd.

[0028] Example 1 Hydrogenated lecithin 12g, cholesterol 6g, DSPE-PEG2000 2g, python oil 3g, safflower extract 1g, Osmanthus fragrans extract 0.5g, madder extract 0.4g, radix lithospermi extract 0.6g, and vitamin E 0.2g are mixed, anhydrous ethanol is added to reach a volume of 100mL to obtain phase A; pearl hydrolysate 0.4g, oyster extract 0.4g, fish collagen peptide 2.4g, glycerol 10g, silk fibroin 0.4g, ergothioneine 0.2g, and vitamin C 0.5g are mixed, PBS solution is added to reach a volume of 100mL to obtain phase B, which is stored at 4℃ for standby.

[0029] First, flush the system with CO2, set the back pressure to 8~10MPa, the temperature to 30℃, and the flow rate to 3mL·min -1 , and stabilize for 5min.

[0030] After the steady state is established, phase A is injected into the "organic phase" inlet of the microfluidic chip at a flow rate of 0.5mL·min -1 , and supercritical CO2 is injected into the "anti-solvent" inlet at a flow rate of 3mL·min -1 ; under the conditions of 30℃ and 8MPa, ethanol is instantaneously diluted and expanded by CO2→lipid is instantaneously supersaturated→<100nm porous lipid microparticles are formed. After staying for 3s, a cyclone separator is directly connected to the chip outlet to collect the dry powder of porous lipid microparticles.

[0031] The collected dry powder is immediately added to phase B at 4℃, and oscillated at 50rpm for 1min, so that the porous lipid microparticles absorb phase B into the interior to form liposomes with a particle size of 80~120nm, which are filtered through a 0.22µm filter membrane to remove bacteria, thereby preparing liposomes with whitening effect.

[0032] Example 2 Take hydrogenated lecithin 11 g, cholesterol 5.6 g, DSPE-PEG2000 1.8 g, python oil 4 g, safflower extract 1 g, osmanthus extract 0.4 g, madder extract 0.4 g, and vitamin E 0.5 g, mix with anhydrous ethanol to a volume of 100 mL to obtain phase A; mix pearl hydrolysate 0.8 g, oyster extract 0.4 g, fish collagen peptide 4 g, glycerol 6 g, silk protein 0.6 g, ergothioneine 0.1 g, and vitamin C 1.6 g, add PBS solution to a volume of 100 mL to obtain phase B, store at 4°C for standby.

[0033] First flush the system with CO2, set the back pressure to 8-10 MPa, temperature 30°C, flow rate 3 mL·min -1 , stable for 5 min.

[0034] After establishing a steady state, inject phase A into the "organic phase" inlet of the microfluidic chip at a rate of 0.5 mL·min -1 , while injecting supercritical CO2 into the "anti-solvent" inlet at a rate of 3 mL·min -1 ; under the conditions of 30°C and 8 MPa, ethanol is instantaneously diluted and expanded by CO2 → lipid is instantaneously supersaturated → <100 nm porous lipid microparticles are formed. After 3 s, a cyclone separator is directly connected to the chip outlet to collect the dry powder of porous lipid microparticles.

[0035] The collected dry powder is immediately added to phase B at 4°C, and oscillated at 50 rpm for 1 min. The porous lipid microparticles absorb phase B into the interior, forming liposomes with a particle size of 80-120 nm. After filtration through a 0.22 µm filter membrane to remove bacteria, liposomes with whitening effect are prepared.

[0036] Example 3 Take hydrogenated lecithin 6 g, cholesterol 3 g, DSPE-PEG2000 1 g, python oil 2 g, safflower extract 0.5 g, osmanthus extract 0.5 g, madder extract 1.5 g, and vitamin E 0.15 g, mix with anhydrous ethanol to a volume of 100 mL to obtain phase A; mix pearl hydrolysate 1 g, oyster extract 1.5 g, fish collagen peptide 1 g, glycerol 6.8 g, silk protein 1 g, ergothioneine 0.05 g, and vitamin C 3 g, add PBS solution to a volume of 100 mL to obtain phase B, store at 4°C for standby.

[0037] First flush the system with CO2, set the back pressure to 8-10 MPa, temperature 30°C, flow rate 3 mL·min -1 , stable for 5 min.

[0038] After establishing a steady state, inject phase A into the "organic phase" inlet of the microfluidic chip at a rate of 0.5 mL·min-1 The "organic phase" inlet of the microfluidic chip was injected, while supercritical CO2 was injected into the "anti-solvent" inlet at a flow rate of 3.5 mL·min -1 The "anti-solvent" inlet was injected; at 30°C and 8 MPa, ethanol was instantaneously diluted and expanded by CO2→ the lipid was instantaneously supersaturated→ <100 nm porous lipid microparticles were formed. After 5 s, a cyclone separator was directly connected to the chip outlet to collect the dry powder of porous lipid microparticles.

[0039] The collected dry powder was immediately added to B phase at 4°C, and oscillated at 50 rpm for 3 min. The porous lipid microparticles absorbed B phase into the interior, forming liposomes with a particle size of 80-120 nm. The liposomes were filtered through a 0.22 µm filter to sterilize, and the liposomes with whitening effect were prepared.

[0040] Comparative Example 1 The difference from Example 1 is that the liposomes were prepared by the traditional thin film-hydration method.

[0041] First, 12 g of hydrogenated lecithin, 6 g of cholesterol, 2 g of DSPE-PEG2000, 3 g of python oil, 1 g of safflower extract, 0.5 g of osmanthus extract, 0.4 g of madder extract, 0.6 g of purple grass extract, and 0.2 g of vitamin E were dissolved in 100 mL of a mixed solvent of chloroform / methanol (volume ratio 9:1) and transferred to a round-bottom flask. Then, rotary evaporation was carried out at 45°C water bath, under vacuum at 0.08-0.09 MPa, at a rotation speed of 80 rpm, until the solvent was completely volatilized, and a uniform and transparent lipid film was formed on the bottle wall; vacuum was continued for 30 min to completely remove the residual solvent.

[0042] Pearl hydrolysate 0.4 g, oyster extract 0.4 g, fish collagen peptide 2.4 g, glycerol 10 g, silk protein 0.4 g, ergothioneine 0.2 g, and vitamin C 0.5 g were mixed, and PBS solution was added to a volume of 100 mL to obtain B' phase, which was stored at 4°C for standby use.

[0043] After the film preparation was completed, 100 mL of B' phase pre-cooled to 4°C was injected into the flask. The flask was placed in a 45°C water bath and rotated at a rotation speed of 60 rpm for 30 min to hydrate under normal pressure, so that the lipid film gradually detached to obtain a milky white crude liposome suspension.

[0044] To obtain liposomes with uniform particle size, the crude suspension was placed in an ice bath and subjected to probe ultrasonic treatment at a power of 200 W: work for 3 s, intermittent for 2 s, and a total time of 5 min; alternatively, an extrusion process can be used, passing through 0.4 µm, 0.2 µm, and 0.1 µm polycarbonate membranes for 5 times each. Finally, the obtained liposome suspension was filtered through a 0.22 µm filter to sterilize, and after dispensing, it was stored at 4°C in the dark to obtain the control liposomes with whitening effect.

[0045] Comparative Example 2 The difference from Example 1 is that vitamin E and ergothioneine are not added.

[0046] Take hydrogenated lecithin 12 g, cholesterol 6 g, DSPE-PEG2000 2 g, python oil 3.2 g, safflower extract 1 g, osmanthus extract 0.5 g, madder extract 0.4 g, and radix lithospermi extract 0.6 g, add anhydrous ethanol to a volume of 100 mL to obtain phase A; mix pearl hydrolysate 0.4 g, oyster extract 0.4 g, fish collagen peptide 2.4 g, glycerol 10.2 g, silk protein 0.4 g, and vitamin C 0.5 g, and add PBS solution to a volume of 100 mL to obtain phase B, which is stored at 4°C for standby.

[0047] First, flush the system with CO2, set the back pressure to 8-10 MPa, the temperature to 30°C, and the flow rate to 3 mL·min-1. -1 Stabilize for 5 min.

[0048] After establishing a steady state, inject phase A into the "organic phase" inlet of the microfluidic chip at a rate of 0.5 mL·min-1. -1 At the same time, inject supercritical CO2 into the "anti-solvent" inlet at a rate of 3 mL·min-1. -1 At 30°C and 8 MPa, the ethanol is instantaneously diluted and expanded by CO2, the lipids are instantaneously supersaturated, and <100 nm porous lipid microparticles are formed. After 3 s, a cyclone separator is directly connected to the chip outlet to collect the dry powder of porous lipid microparticles.

[0049] The collected dry powder is immediately added to phase B at 4°C, and oscillated at 50 rpm for 1 min. The porous lipid microparticles absorb phase B into the interior, forming liposomes with a particle size of 80-120 nm. The liposomes are filtered through a 0.22 µm filter membrane to sterilize, and the liposomes with whitening effect are prepared.

[0050] Comparative Example 3 The difference from Example 1 is that osmanthus extract is not used.

[0051] Take hydrogenated lecithin 12 g, cholesterol 6 g, DSPE-PEG2000 2 g, python oil 3.5 g, safflower extract 1 g, madder extract 0.4 g, radix lithospermi extract 0.6 g, and vitamin E 0.2 g, add anhydrous ethanol to a volume of 100 mL to obtain phase A; mix pearl hydrolysate 0.4 g, oyster extract 0.4 g, fish collagen peptide 2.4 g, glycerol 10 g, silk protein 0.4 g, ergothioneine 0.2 g, and vitamin C 0.5 g, and add PBS solution to a volume of 100 mL to obtain phase B, which is stored at 4°C for standby.

[0052] First flush the system with CO2, set back pressure 8~10 MPa, temperature 30℃, flow rate 3 mL·min -1 , stable for 5 min.

[0053] After establishing a steady state, inject A phase into the "organic phase" inlet of the microfluidic chip at a rate of 0.5 mL·min -1 , while injecting supercritical CO2 into the "antisolvent" inlet at a rate of 3 mL·min -1 ; under the conditions of 30℃ and 8 MPa, ethanol is instantaneously diluted and expanded by CO2→lipid is instantaneously supersaturated→<100 nm porous lipid microparticles are formed. After staying for 3 s, a cyclone separator is directly connected to the chip outlet to collect the dry powder of porous lipid microparticles.

[0054] The collected dry powder is immediately added to B phase at 4℃, and oscillated at 50 rpm for 1 min. The porous lipid microparticles absorb B phase into the interior, forming liposomes with a particle size of 80~120 nm. After filtering through a 0.22 µm filter membrane to sterilize, liposomes with whitening efficacy are prepared.

[0055] Comparative Example 4 The difference from Example 1 is that fish collagen peptide and silk protein are not added.

[0056] Take 12 g of hydrogenated lecithin, 6 g of cholesterol, 2 g of DSPE-PEG2000, 3 g of python oil, 1 g of safflower extract, 0.5 g of osmanthus extract, 0.4 g of madder extract, 0.6 g of purple grass extract, and 0.2 g of vitamin E, mix them, add anhydrous ethanol to a volume of 100 mL to obtain A phase; mix 0.4 g of pearl hydrolysate, 0.4 g of oyster extract, 12.8 g of glycerol, 0.2 g of ergothioneine, and 0.5 g of vitamin C, add PBS solution to a volume of 100 mL to obtain B phase, and store at 4℃ for standby.

[0057] First flush the system with CO2, set back pressure 8~10 MPa, temperature 30℃, flow rate 3 mL·min -1 , stable for 5 min.

[0058] After establishing a steady state, inject A phase into the "organic phase" inlet of the microfluidic chip at a rate of 0.5 mL·min -1 , while injecting supercritical CO2 into the "antisolvent" inlet at a rate of 3 mL·min -1 ; under the conditions of 30℃ and 8 MPa, ethanol is instantaneously diluted and expanded by CO2→lipid is instantaneously supersaturated→<100 nm porous lipid microparticles are formed. After staying for 3 s, a cyclone separator is directly connected to the chip outlet to collect the dry powder of porous lipid microparticles.

[0059] The collected dry powder was immediately added to B phase at 4°C, and oscillated at 50 rpm for 1 min, and the porous lipid microparticles absorbed B phase into the interior to form liposomes with particle sizes of 80-120 nm. The liposomes were filtered through a 0.22 μm filter membrane to remove bacteria, and the liposomes with whitening efficacy were prepared.

[0060] Test Example 1 B16 cells were inoculated in a 96-well plate at a density of 1 × 10 4 cells per well, and cultured at 37°C in 5% CO2 for 24 h to allow the cells to adhere. Then, the cells were divided into groups 1-7, a blank control group, and a positive control group, with 3 replicate wells for each group. After the original culture solution was discarded, 190 μL of culture solution was added to each well, and then active ingredients were added to each well as follows: Blank control group: 10 μL of a PBS solution was added to each well.

[0061] Positive control group: 10 μL of a nicotinamide solution was added to each well to give a final concentration of 2 mM.

[0062] Group 1: 10 μL of the liposomes with whitening efficacy prepared in Example 1 were added to each well.

[0063] Group 2: 10 μL of the liposomes with whitening efficacy prepared in Example 2 were added to each well.

[0064] Group 3: 10 μL of the liposomes with whitening efficacy prepared in Example 3 were added to each well.

[0065] Group 4: 10 μL of the liposomes with whitening efficacy prepared in Comparative Example 1 were added to each well.

[0066] Group 5: 10 μL of the liposomes with whitening efficacy prepared in Comparative Example 2 were added to each well.

[0067] Group 6: 10 μL of the liposomes with whitening efficacy prepared in Comparative Example 3 were added to each well.

[0068] Group 7: 10 μL of the liposomes with whitening efficacy prepared in Comparative Example 4 were added to each well.

[0069] After continued culture for 72 h, the reaction substrate was added according to the instructions of a tyrosinase activity assay kit, and after incubation at 37°C for a certain period of time, the stop solution was added. The absorbance was measured at 490 nm using an enzyme marker, and the tyrosinase activity inhibition rate was calculated.

[0070]

[0071] Table 1: Tyrosinase activity inhibition rate of the composition

[0072] As can be seen from Table 1, compared with a single whitening ingredient, the composition provided by the present application can effectively inhibit tyrosinase activity and has good whitening effect. The liposome prepared by the method provided by the present application can make the active ingredient more stable and easier to absorb, thereby further improving the whitening effect.

[0073] Test Example 2 The liposomes prepared in Examples 1-3 and Comparative Example 1 were used as samples, and vitamin E and vitamin C were used as markers to detect the encapsulation rate thereof in the liposomes. The encapsulation rate of vitamin E was determined by Sephadex G-50 microcolumn centrifugation-HPLC method, and the encapsulation rate of vitamin C was determined by 10 kDa ultrafiltration centrifugation-HPLC method. The results are shown in Table 2.

[0074] Table 2: Encapsulation rate of liposomes

[0075] As can be seen from Table 2, compared with the traditional film-hydration method, the method provided by the present application can effectively improve the encapsulation rate of the liposomes.

[0076] The liposomes prepared in Examples 1 and Comparative Example 1 were evenly packed in brown centrifuge tubes and stored at 4°C in the dark to avoid repeated freeze-thawing to reduce external interference. The stability of the samples was detected at 0 days, 14 days and 30 days, respectively. The appearance change was observed by naked eye and photographed. After the sample was broken by methanol, it was ultrasonically treated and centrifuged to obtain the supernatant, which was filtered through a 0.22 μm filter membrane and then analyzed by sample injection. The retention rate of VC at different time points was calculated to verify the retention effect of the active ingredient, and the tyrosinase inhibition rate was determined according to the method described in Test Example 1. The results are shown in Table 3.

[0077] Table 3: Results of liposome stability test

[0078] As can be seen from Table 3, the liposomes prepared by the present application had no obvious change in appearance after being stored at 4°C for 30 days, the active ingredient (VC) content and activity retention rate were more than 90%, and the tyrosinase inhibition activity retention rate was as high as 94.53%. This indicates that the liposomes prepared by the present application still have good stability after being stored at 4°C in the dark for 30 days.

[0079] Test Example 3 The liposomes prepared in Example 1 were used to prepare a whitening cream according to the component ratio in the following table. The specific preparation method was as follows: Stearic acid, white vaseline and glycerol monostearate were heated to 80°C to completely melt to obtain an oil phase; glycerol, triethanolamine and preservatives were dissolved in distilled water and heated to 80°C to obtain an aqueous phase.

[0080] The water phase is slowly added to the oil phase under stirring, and the stirring is continued until a uniform emulsion is formed; when the temperature drops below 40℃, the liposome suspension is slowly added, and the stirring is continued until room temperature. After dispensing, the product is stored at 4℃ in the dark, as shown in Figure 1

[0081] Table 4: Proportion of whitening cream and blank cream

[0082] The blank cream is prepared according to the proportion in Table 4 as a control: stearic acid, white petrolatum, and glycerol monostearate are heated to 80℃ to completely melt, to obtain an oil phase; glycerol, triethanolamine, and preservatives are dissolved in distilled water, and heated to 80℃ to obtain a water phase. The water phase is slowly added to the oil phase under stirring, and the stirring is continued until a uniform emulsion is formed; after the temperature drops to room temperature, the product is stored at 4℃ in the dark after dispensing.

[0083] After the whitening cream and the blank cream are prepared, volunteers are recruited for a whitening test.

[0084] A total of 30 volunteers are recruited for the test, aged 18-50 years, with mild to moderate uneven skin tone, dullness, or post-sun pigmentation on the face, without serious skin diseases, allergic history, and without using other whitening products in the past month. After skin testing, the volunteers do not have an irritating reaction to the whitening cream prepared by the application. All volunteers have voluntarily signed an informed consent form before the test.

[0085] Each volunteer applies the whitening cream prepared by the application and the blank cream without added liposomes to the left and right halves of the face, respectively, once in the morning and once in the evening, for 12 weeks.

[0086] At 0, 4, 8, and 12 weeks, the skin parameters of the left and right faces of the volunteers are detected. The L* value of the skin is measured using a colorimeter, and the facial melanin content (MI, VISIA-CR arbitrary unit) is quantified using a Visia-CR skin analyzer. The melanin reduction rate is calculated, and the changes in the area of pigmented spots, contrast, and skin tone uniformity are analyzed. The results are shown in Table 5.

[0087]

[0088] Wherein MI initial represents the facial melanin content of the volunteer at 0 weeks; and MI end represents the facial melanin content of the volunteer at each measurement.

[0089] Table 5: Whitening effect of volunteers

[0090] As shown in Table 5, after applying the whitening cream prepared by the application to the left face of the volunteer, the skin melanin content is effectively reduced, and the area of pigmented spots is reduced, showing a good whitening effect. ​

[0091] Although the above embodiments have been described in detail, they are only some embodiments of the present application, not all embodiments. Other embodiments can be obtained on the basis of the above embodiments without creativity, which are within the protection scope of the present application.

Claims

1. A plant extract composition having a whitening effect, characterized by, The composition comprises python oil 1-5 parts by mass, safflower extract 0.5-3 parts by mass, osmanthus extract 0.1-2 parts by mass, pearl hydrolysate 0.5-2 parts by mass, oyster extract 0.3-1.5 parts by mass, madder extract 0.3-1.5 parts by mass, comfrey extract 0.2-1 part by mass, fish collagen peptide 1-4 parts by mass, glycerol 5-12 parts by mass, silk fibroin 0.2-1 part by mass, ergothioneine 0.02-0.2 parts by mass, vitamin C 0.5-3 parts by mass, and vitamin E 0.1-1 part by mass.

2. The method of claim 1, wherein, The content of hydroxysafflor yellow A in the safflower extract is 5%wt-10%wt; the content of verbascoside in the osmanthus extract is 15%wt-30%wt; the content of alizarin in the madder extract is ≥5%wt; and the content of comfrey in the comfrey extract is 2%wt-5%wt.

3. The method of claim 1, wherein, The average molecular weight of the fish collagen peptide is ≤1000Da, and the content of hydroxyproline is ≥8%wt; the content of hydrolyzed silk fibroin in the silk fibroin is ≥90%wt, and the molecular weight of the silk fibroin is 500-3000Da.

4. Liposomes with whitening efficacy, characterized in that, The liposome comprises the composition of claim 1.

5. A method of preparing the liposome of claim 4, characterized in that, The method comprises the following steps: dissolving film-forming lipids, python oil, safflower extract, osmanthus extract, madder extract, comfrey extract, and vitamin E in anhydrous ethanol to obtain phase A; dissolving pearl hydrolysate, oyster extract, fish collagen peptide, glycerol, silk fibroin, ergothioneine, and vitamin C in a PBS solution to obtain phase B; A phase is injected into the organic phase inlet of the microfluidic chip at a flow rate of 0.4-0.6 mL·min -1 Supercritical CO2 is injected into the anti-solvent inlet of the microfluidic chip at a flow rate of 2.5-3.5 mL·min -1 The anti-solvent inlet of the microfluidic chip is injected; the porous lipid microparticles are formed under the conditions of 25-30 ℃ and 7-15 MPa, and the porous lipid microparticle dry powder is collected at the chip outlet after 2-5 s of residence. adding phase B to the dry powder of porous lipid microparticles, shaking well to form liposomes; the film-forming lipids are a mixture of hydrogenated lecithin, cholesterol, and distearoyl phosphatidyl ethanolamine-polyethylene glycol in a mass ratio of 5.5-6.5:2.8-3.2:0.9-1.

1.

6. The method of claim 5, wherein, 5mL of phase A and 5mL of phase B are prepared per g of the composition of claim 1, and 1-2g of film-forming lipids are used per g of the composition of claim 1.

7. The method of claim 5, wherein, The particle size of the porous lipid microparticles is <100nm.

8. The method of claim 5, wherein, The temperature of phase B is 2-8°C.

9. The method of claim 5, wherein, The shaking condition is 40-80rpm oscillation for 1-3min.

10. A whitening cream, characterized by, The whitening cream contains the liposome of claim 4.

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