Preparation method of cacumen biotae extract, related product and application

Abstract

The invention provides a preparation method of a cacumen biotae extract, a related product and application, and belongs to the technical field of daily chemicals. The preparation method comprises the following steps: S1, mixing cacumen biotae with water, carrying out circulating percolation extraction, and combining filtrate to obtain cacumen biotae circulating percolate and medicine residues; s2, the cacumen biotae circulating leachate obtained in the step S1 is subjected to filter membrane concentration, concentration and filtration, and cacumen biotae concentrated filtrate is obtained; s3, carrying out reduced pressure distillation and concentration on the cacumen biotae concentrated filtrate obtained in the step S2, and synchronously collecting a concentrated solution and a condensate obtained by reduced pressure distillation; s4, cooling the concentrated solution obtained in the step S3, mixing the concentrated solution with an ethanol water solution to obtain a mixed solution, cooling, standing, filtering, filtering with a molecular entrapment membrane, concentrating and drying to obtain a cacumen biotae extract; s5, the medicine residues obtained in the step S1 are ground into thick liquid, and the platycladus orientalis thick liquid is obtained. The cacumen biotae extract prepared by the method can be compatible with different daily chemical product systems, and has a good effect of regulating and controlling skin water-oil balance.

Description

Technical Field

[0001] The invention belongs to the technical field of daily chemical products and relates to a preparation method of a Platycladus orientalis leaf extract, related products and applications. Background Art

[0002] The skin barrier is often described as a "brick wall structure", in which keratinocytes represent "bricks" and intercellular lipids represent "mortar". The "mortar" is mainly polar lipids synthesized by keratinocytes, and its main function is to reduce water loss and maintain the skin barrier; the sebum synthesized by sebaceous gland cells is mainly neutral lipids, which are secreted to the surface of the skin and form a sebum film on the outer layer of the epidermis. The sebaceous glands will gradually mature with age and gradually degenerate with the aging process, but babies can have the smoothest and healthiest skin, while the skin of the elderly is often dry and wrinkled. This is because the keratinocytes of babies are highly active, and the intercellular lipids they synthesize are highly hydrophilic and can firmly lock the moisture of the epidermis. The keratinocytes of the elderly gradually age and lose cell vitality with age, and the ability to synthesize intercellular lipids decreases, making the epidermal moisture very easy to lose. This shows that the epidermal moisturizing ability and skin barrier function are highly correlated with keratinocytes, but less correlated with sebaceous gland function.

[0003] Sebum secreted by sebaceous glands is mainly composed of non-polar lipids, such as triglycerides, wax esters and squalene, while polar lipids synthesized by keratinocytes are mainly a mixture of ceramides, free fatty acids and cholesterol. As the age enters puberty, the sebaceous glands mature and their activities become active. Acne often occurs in areas rich in sebaceous glands, but the proportion of lipids produced by keratinocytes in the total amount of extractable surface lipids is negligible. This shows that in the pathophysiological process of acne, sebum secreted by sebaceous glands is more involved, while lipids synthesized by keratinocytes are less involved. Therefore, improving the ability of keratinocytes to synthesize intercellular lipids and reducing the content of sebum synthesized by sebaceous gland cells is beneficial to maintaining water and oil balance on the skin surface.

[0004] Platycladus orientalis leaf, also known as cypress leaf, cypress leaf, and cypress leaf, is a plant of the genus Platycladus orientalis in the Cupressaceae family. Platycladus Oriental (L.) Franco Thuja orientalis L.; Biota orientalis(L.) Endl.]) branches and leaves. Its branches vary in length, are multi-branched, and the twigs are flat. The leaves are small and scaly, alternately arranged, attached to the branches, and are dark green or yellow-green. They are brittle and easy to break. They have a fresh aroma and taste bitter and slightly spicy. The leaves of Platycladus orientalis contain volatile oils, fatty acids, flavonoids, etc., including α-thujone, thujene, fenchone, pinene, caryophyllene, palmitic acid, stearic acid, lauric acid, capric acid, quercetin, apigenin, cedarwood bioflavonoids, kaempferol-7-O-rhamnoside, myricetin-3-O-rhamnoside, myricetin, cypress bioflavonoids, tachyphyllotoxin bioflavonoids, pimaric acid, isopimaric acid, sandaracine, 10-nonacosanol, β-sitosterol, condensed tannins, deoxypodophyllotoxin, etc. Traditional medicine believes that Platycladus orientalis leaves have bactericidal and hemostatic effects, and can be used externally to treat hair loss, mumps, burns and other diseases.

[0005] Chinese invention patent CN117643560A provides a Platycladus orientalis extract and its preparation method and application. Platycladus orientalis wood and Platycladus orientalis leaves are used as raw materials, and sorbitol dehydrogenase, retinol dehydrogenase and phosphite dehydrogenase are added after water extraction and stirred to obtain Platycladus orientalis extract fermentation liquid. Platycladus orientalis extract fermentation liquid is prepared with cholesterol, caprylic / capric triglyceride, polyglycerol-10 diisostearate, etc. to prepare nanoemulsion. The Platycladus orientalis extract fermentation liquid of this technology has antibacterial properties and is used in hair care and shampoo cosmetics to reduce scalp oil secretion, stabilize the fullness and health of hair follicles, and inhibit hair loss.

[0006] Chinese invention patent CN118986805A provides an extract of Platycladus orientalis leaves for preventing hair loss, and its preparation method and application. The effective ingredients in Platycladus orientalis leaves are extracted by alkali extraction and acid precipitation, and no organic solvent is used, which is environmentally friendly. Under the premise of ensuring efficacy, the safety of production is improved and the difficulty of purification is reduced. Disodium ethylenediaminetetraacetic acid, ascorbic acid or sodium citrate are used as protective agents to avoid the use of borax, which on the one hand increases the yield of total flavonoids and on the other hand eliminates potential risks, making the Platycladus orientalis leaf extract meet the safety requirements of the cosmetics industry. The Platycladus orientalis leaf extract used for preventing hair loss acts on the hair follicles, increases the average length of the hair follicles, and increases the VEGF content, so it has a significant anti-hair loss effect.

[0007] Chinese invention patent CN117717500A provides a method for extracting Platycladus orientalis leaf extract, comprising the following steps: step (1), soaking Platycladus orientalis leaf in water, then adding composite bacteria and cultivating for 4-6 days; step (2), heating and extracting; step (3), filtering and obtaining filtrate; step (4), concentrating and drying the filtrate to obtain Platycladus orientalis leaf extract; the composite bacteria is a combination of aquatic Rahnella, Leuconostoc lactis and acetic acid bacteria. This technology has the advantage of improving the efficacy of Platycladus orientalis leaf extract.

[0008] However, the composition of Platycladus orientalis leaf extract in the prior art is relatively complex, which is not conducive to smooth compatibility with the various ingredients in daily chemical products of different systems; in addition, due to the variety of effective ingredients contained in Platycladus orientalis leaves, the efficacy and compatibility of Platycladus orientalis leaf extracts prepared by different preparation methods with daily chemical products are often difficult to predict. The prior art has not yet provided a Platycladus orientalis leaf extract and its preparation process that has better effect on regulating skin oil-water balance and stronger compatibility with different cosmetic systems. Summary of the invention

[0009] In view of this, in order to solve the problems existing in the above-mentioned prior art, the object of the present invention is to provide a preparation method of Platycladus orientalis leaf extract, related products and applications.

[0010] To achieve the above-mentioned object of the invention, on the one hand, the present invention provides a method for preparing a Platycladus orientalis leaf extract, comprising the following steps: S1. Mix the Platycladus orientalis leaves with 4-16 times the weight of water, perform cyclic filtration extraction, repeat 2-4 times, combine the filtrates, and obtain the Platycladus orientalis leaf cyclic filtrate and medicinal residues; S2, performing membrane concentration on the Platycladus orientalis leaf circulating leachate obtained in step S1, concentrating the leachate to 5-7 times the weight of the Platycladus orientalis leaf in step S1, filtering, and obtaining Platycladus orientalis leaf concentrated filtrate; S3, performing vacuum distillation to concentrate the concentrated filtrate of Platycladus orientalis leaves obtained in step S2, and simultaneously collecting the concentrated solution and condensate obtained by the vacuum distillation, wherein the condensate is Platycladus orientalis hydrosol; S4, cooling the concentrated solution obtained in step S3, mixing it with ethanol aqueous solution to obtain a mixed solution, cooling it to -6~6°C and letting it stand, filtering it, filtering it with a molecular retention membrane, concentrating it, and drying it to obtain a Platycladus orientalis leaf extract; S5, grinding the residue obtained in step S1 to obtain Platycladus orientalis slurry.

[0011] Preferably, in step S1, the mass content of quercetin in the Platycladus orientalis leaves is ≥ 0.25%, more preferably ≥ 0.3%.

[0012] Preferably, in step S1, each cyclic diafiltration extraction comprises the following steps: cyclic diafiltration at 80-99°C for 30-90 min, filtering, and cooling the filtrate to ≤60°C.

[0013] More preferably, in step S1, the cyclic diafiltration extraction is repeated 3 times. The first extraction comprises the following steps: mixing Platycladus orientalis leaves with 10 times the weight of water, performing cyclic percolation extraction at 90-99° C. for 55-65 minutes, filtering, rapidly cooling the filtrate to below 60° C., and collecting the filter residue; The second extraction comprises the following steps: adding 6 times of water to the residue obtained in the first extraction step, performing cyclic percolation extraction at 90-99°C for 55-65 minutes, filtering, rapidly cooling the filtrate to below 60°C, and collecting the residue; The third extraction comprises the following steps: adding 4 times of water to the filter residue obtained in the first extraction step, cyclic percolation extraction at 90-99° C. for 55-65 minutes, filtering, rapidly cooling the filtrate to below 60° C., and collecting the filter residue.

[0014] Preferably, in step S2, the membrane concentration is concentration using a 100-300 Dalton membrane, and the filtration is filtration until clarified.

[0015] More preferably, in step S2, the membrane concentration is performed using a 100 Dalton membrane.

[0016] Preferably, in step S3, the temperature of the reduced pressure distillation concentration is ≤70°C, and the weight of the concentrated liquid is 0.8-1.4 times the weight of the Platycladus orientalis leaves in step S1, and more preferably 1-1.2 times.

[0017] Preferably, in step S4, the volume fraction of ethanol in the ethanol aqueous solution is 90%-99%, more preferably 95%; the volume fraction of ethanol in the mixed solution is 20%-30%, more preferably 25%.

[0018] Preferably, in step S4, the standing time is ≥ 2 h.

[0019] More preferably, and as an example of the present invention, the standing time is 2 hours.

[0020] Preferably, in step S4, the temperature is lowered to 0-4°C and allowed to stand.

[0021] Preferably, the molecular cut-off membrane filtration is filtration using a molecular cut-off membrane of 1000-5000 Daltons.

[0022] More preferably, the molecular cut-off membrane filtration is filtration using a molecular cut-off membrane of 2000 Daltons.

[0023] Preferably, in step S5, the refining is specifically refining with a colloid mill, and the refining is performed to a solid average particle size of 10-30 μm.

[0024] Preferably, the preparation method further comprises step S601: S601, drying the Platycladus orientalis slurry obtained in step S5 to obtain Platycladus orientalis leaf powder.

[0025] Preferably, the preparation method further comprises step S602: S602, concentrating the Platycladus orientalis slurry obtained in step S5 to a solid weight content of 20%-35% to obtain Platycladus orientalis leaf pulp.

[0026] On the other hand, the present invention provides a Platycladus orientalis leaf extract obtained by the above preparation method.

[0027] On the other hand, the present invention provides Platycladus orientalis hydrosol obtained by the above-mentioned preparation method.

[0028] On the other hand, the present invention provides Platycladus orientalis slurry obtained by the above preparation method.

[0029] On the other hand, the present invention provides Platycladus orientalis leaf powder obtained by the above-mentioned preparation method.

[0030] On the other hand, the present invention provides Platycladus orientalis leaf pulp prepared by the above-mentioned preparation method.

[0031] In another aspect, the present invention provides application of the above preparation method in producing Platycladus orientalis leaf extract.

[0032] On the other hand, the present invention provides the use of the Platycladus orientalis leaf extract, Platycladus orientalis hydrosol, Platycladus orientalis leaf powder, Platycladus orientalis slurry and Platycladus orientalis leaf slurry obtained by the above-mentioned preparation method in cosmetics.

[0033] Preferably, the cosmetic is selected from at least one of the following: (1) Cosmetics that inhibit sebaceous gland oil secretion; (2) Cosmetics that promote the expression of FLG (filagrin gene) and Caspase14 in keratinocytes; (3) Cosmetics that inhibit oil secretion and promote oil-water balance; (4) Cosmetics that treat acne, prevent acne, replenish moisture, and inhibit oil secretion.

[0034] More preferably, the cosmetic is: (1) Cosmetics that inhibit sebaceous gland oil secretion; (2) Cosmetics that promote the expression of FLG and Caspase14 in keratinocytes; (3) Cosmetics that inhibit oil secretion and promote oil-water balance; (4) Cosmetics that treat acne, prevent acne, replenish moisture, and inhibit oil secretion.

[0035] Preferably, the cosmetic is a cosmetic that promotes the expression of at least one gene among ELOVL4 (human elongation factor very long chain fatty acid-like protein 4), ELOVL7 (human elongation factor very long chain fatty acid-like protein 7), CerS4 (ceramide synthase 4), inhibits oil secretion, and promotes oil-water balance.

[0036] On the other hand, the present invention provides a cosmetic, the ingredients of which include the Platycladus orientalis leaf extract, Platycladus orientalis hydrosol, Platycladus orientalis leaf powder, Platycladus orientalis leaf pulp, and Platycladus orientalis pulp that promote oil-water balance and are prepared by the above-mentioned preparation method.

[0037] In another aspect, the present invention provides a shampoo comprising the following ingredients: Water, sodium laureth sulfate, laureth-2, sodium sulfate, Platycladus orientalis leaf extract, glycerin, polyquaternium-10, sodium cocoamidopropionate, sodium chloride, cocamidopropyl betaine, citric acid, cocamide MEA, phenoxyethanol and pH adjuster; Wherein, the Platycladus orientalis leaf extract is the Platycladus orientalis leaf extract prepared by the above-mentioned preparation method.

[0038] Preferably, the shampoo comprises the following ingredients by weight percentage: Sodium Laureth Sulfate 9%-10%, Laureth-2 0.1%-0.5%, Sodium Sulfate 0.01%-0.2%, Platycladus Orientalis Leaf Extract 0.2%-1%, Glycerin 1%-5%, Polyquaternium-10 0.1%-1%, Sodium Cocoamidopropionate 0.5%-0.1%, Sodium Chloride 0.2%-0.5%, Cocamidopropyl Betaine 4.5%-6.5%, Citric Acid 0.001%-1%, Cocamide MEA 0.1%-0.8%, Phenoxyethanol 0.1%-1%, Sodium Hydroxide 0.001%-0.1% and Water.

[0039] More preferably, the shampoo comprises the following ingredients by weight percentage: Sodium laureth sulfate 9.18%-9.72%, laureth-2 0.135%-0.4725%, sodium sulfate 0.0675%-0.135%, Platycladus orientalis leaf extract 0.5%, glycerin 3%, polyquaternium-10 0.2%-0.4%, sodium cocoamidopropionate 0.84%-0.9%, sodium chloride 0.403%-0.43%, cocamidopropyl betaine 5.45%, citric acid 0.05%, cocamide MEA 0.4%, phenoxyethanol 0.7%, sodium hydroxide 0.005% and water.

[0040] More preferably, the shampoo consists of the following ingredients by weight percentage: Sodium laureth sulfate 9.18%-9.72%, laureth-2 0.135%-0.4725%, sodium sulfate 0.0675%-0.135%, Platycladus orientalis leaf extract 0.5%, glycerin 3%, polyquaternium-10 0.2%-0.4%, sodium cocoyl propionate 0.84%-0.9%, sodium chloride 0.403%-0.43%, cocamidopropyl betaine 5.45%, citric acid 0.05%, cocamide MEA 0.4%, phenoxyethanol 0.7%, sodium hydroxide 0.005% and the rest water.

[0041] As an example of the present invention, the shampoo is composed of the following ingredients by weight percentage: Sodium laureth sulfate 9.18%-9.72%, laureth-2 0.135%-0.4725%, sodium sulfate 0.0675%-0.135%, Platycladus orientalis leaf extract 0.5%, glycerin 3%, polyquaternium-10 0.4%, sodium cocoyl propionate 0.84%-0.9%, sodium chloride 0.403%-0.43%, cocamidopropyl betaine 5.45%, citric acid 0.05%, cocamide MEA 0.4%, phenoxyethanol 0.7%, sodium hydroxide 0.005% and the rest water.

[0042] As an example of the present invention, the shampoo is composed of the following ingredients by weight percentage: Sodium laureth sulfate 9.18%-9.72%, laureth-2 0.135%-0.4725%, sodium sulfate 0.0675%-0.135%, Platycladus orientalis leaf extract 0.5%, glycerin 3%, polyquaternium-10 0.2%, sodium cocoyl propionate 0.84%-0.9%, sodium chloride 0.403%-0.43%, cocamidopropyl betaine 5.45%, citric acid 0.05%, cocamide MEA 0.4%, phenoxyethanol 0.7%, sodium hydroxide 0.005% and the rest water.

[0043] As an example of the present invention, the shampoo is composed of the following ingredients by weight percentage: Sodium laureth sulfate 9.18%-9.72%, laureth-2 0.135%-0.4725%, sodium sulfate 0.0675%-0.135%, Platycladus orientalis leaf extract 0.5%, glycerin 3%, polyquaternium-10 0.3%, sodium cocoyl propionate 0.84%-0.9%, sodium chloride 0.403%-0.43%, cocamidopropyl betaine 5.45%, citric acid 0.05%, cocamide MEA 0.4%, phenoxyethanol 0.7%, sodium hydroxide 0.005% and the rest water.

[0044] In the shampoo, each component can be provided separately or simultaneously from existing single-component raw materials and compound raw materials.

[0045] As an example of the present invention, in the shampoo, the sodium laureth sulfate, laureth-2 and sodium sulfate are provided by RSAW ESB70 / ZA.

[0046] As an example of the present invention, in the shampoo, the sodium cocoamide propionate is provided by OULI 115L.

[0047] As an example of the present invention, in the shampoo, the cocamidopropyl betaine and citric acid are provided by TEGO BETAIN C60.

[0048] As an example of the present invention, in the shampoo, the polyquaternium-10 is prepared by UCARE TM Polymer JR-400 is provided.

[0049] As an example of the present invention, in the shampoo, the cocamide is provided by Comperlan 100C.

[0050] As an example of the present invention, in the shampoo, the phenoxyethanol is provided by Microcare PE.

[0051] Preferably, the shampoo consists of the following ingredients by weight: RSAW ESB70 / ZA 13.5%, Platycladus orientalis leaf extract 5%, Glycerin 3%, UCARE TM Polymer JR-400 0.2%-0.4%, OULI-115L 3%, TEGO BETAIN C60 10%, Comperlan 100C 0.4%, Microcare PE 0.7%, sodium hydroxide 0.005%, sodium chloride 0.4% and the balance water.

[0052] As an example of the present invention, the shampoo is composed of the following ingredients by weight percentage: RSAW ESB70 / ZA 13.5%, Platycladus orientalis leaf extract 5%, Glycerin 3%, UCARE TM Polymer JR-400 0.2%, OULI-115L 3%, TEGO BETAIN C60 10%, Comperlan 100C 0.4%, Microcare PE 0.7%, sodium hydroxide 0.005%, sodium chloride 0.4% and the balance water.

[0053] As an example of the present invention, the shampoo is composed of the following ingredients by weight percentage: RSAW ESB70 / ZA 13.5%, Platycladus orientalis leaf extract 5%, Glycerin 3%, UCARE TM Polymer JR-400 0.3%, OULI-115L 3%, TEGO BETAIN C60 10%, Comperlan 100C 0.4%, Microcare PE 0.7%, sodium hydroxide 0.005%, sodium chloride 0.4% and the balance water.

[0054] More preferably, and as an example of the present invention, the shampoo consists of the following ingredients by weight percentage: RSAW ESB70 / ZA 13.5%, Platycladus orientalis leaf extract 5%, Glycerin 3%, UCARE TM Polymer JR-400 0.4%, OULI-115L 3%, TEGO BETAIN C60 10%, Comperlan 100C 0.4%, Microcare PE 0.7%, sodium hydroxide 0.005%, sodium chloride 0.4% and the balance water.

[0055] In another aspect, the present invention provides a method for preparing the above shampoo, comprising the following steps: (1) Heat water to 80-85°C and mix with formula amount of RSAW ESB70 / ZA to obtain phase A; (2) Mixing the Platycladus orientalis leaf extract, two-thirds of the formula amount of glycerin and water to obtain phase B; (3) UCARE TM Polymer JR-400 is mixed with the remaining glycerin, then mixed with water, and heated to 80-85°C to obtain phase C; (4) OULI-115L, TEGO BETAIN C60, Comperlan 100C and Microcare PE were mixed to obtain phase D; (5) Mix sodium hydroxide, sodium chloride and water to obtain phase E; (6) Mix phase A, phase B, phase C and phase D in sequence, then add phase E and adjust the pH value to 5.5-7 to obtain shampoo.

[0056] In another aspect, the present invention provides a scalp essence comprising the following ingredients: Water, polyacrylate crosspolymer-6, Platycladus orientalis leaf extract, 1,2-hexanediol, butylene glycol, sodium metabisulfite, and arginine; Wherein, the Platycladus orientalis leaf extract is the Platycladus orientalis leaf extract prepared by the above-mentioned preparation method.

[0057] Preferably, the scalp essence comprises the following ingredients by weight percentage: Polyacrylate crosspolymer-6 0.05%-0.5%, Platycladus orientalis leaf extract 0.1%-1%, 1,2-hexanediol 0.1%-20%, butylene glycol 0.01%-20%, sodium metabisulfite 0.01%-0.5%, arginine 0.01%-0.5% and water.

[0058] More preferably, the scalp essence comprises the following ingredients by weight percentage: Polyacrylate Crosspolymer-6 0.2%, Platycladus Orientalis Leaf Extract 0.5%, 1,2-Hexanediol 2%, Butylene Glycol 0.01%-20%, Sodium Metabisulfite 0.05%, Arginine 0.1% and Water.

[0059] As an example of the present invention, the scalp essence is composed of the following ingredients by weight percentage: Polyacrylate crosspolymer-6 0.2%, Platycladus orientalis leaf extract 0.5%, 1,2-hexanediol 2%, butylene glycol 5%, sodium metabisulfite 0.05%, arginine 0.1% and the balance water.

[0060] As an example of the present invention, the scalp essence is composed of the following ingredients by weight percentage: Polyacrylate crosspolymer-6 0.2%, Platycladus orientalis leaf extract 0.5%, 1,2-hexanediol 2%, butylene glycol 10%, sodium metabisulfite 0.05%, arginine 0.1% and the balance water.

[0061] As an example of the present invention, the scalp essence is composed of the following ingredients by weight percentage: Polyacrylate crosspolymer-6 0.2%, Platycladus orientalis leaf extract 0.5%, 1,2-hexanediol 2%, butylene glycol 20%, sodium metabisulfite 0.05%, arginine 0.1% and the balance water.

[0062] More preferably, and as an example of the present invention, the scalp essence is composed of the following ingredients by weight percentage: Polyacrylate crosspolymer-6 0.2%, Platycladus orientalis leaf extract 0.5%, 1,2-hexanediol 2%, butylene glycol 2%, sodium metabisulfite 0.05%, arginine 0.1% and the balance water.

[0063] In another aspect, the present invention provides a method for preparing the scalp essence, comprising the following steps: (1) Dissolve polyacrylate cross-linked polymer-6 in water, heat to 80-85°C, stir evenly, and cool to 50°C to obtain phase A; (2) mixing and dissolving the Platycladus orientalis leaf extract, 1,2-hexanediol, butanediol, sodium pyrosulfite, arginine and water to obtain phase B; (3) Mix phase A and phase B to obtain scalp essence.

[0064] In another aspect, the present invention provides a Platycladus orientalis leaf mud film, comprising the following components: Water, polyacrylate crosspolymer-6, 1,2-hexanediol, p-hydroxyacetophenone, arginine, glycerin and Thuja orientalis leaf pulp; Wherein, the Platycladus orientalis leaf extract is Platycladus orientalis leaf pulp prepared by the above-mentioned preparation method.

[0065] Preferably, the Platycladus orientalis leaf mud film comprises the following components by weight percentage: Polyacrylate crosspolymer-6 0.1%-2%, 1,2-hexanediol 0.1%-1%, p-hydroxyacetophenone 0.1%-1%, arginine 0.01%-1%, glycerin 1%-10%, Thuja orientalis leaf pulp 70%-90% and water.

[0066] More preferably, the Platycladus orientalis leaf mud film comprises the following components by weight percentage: Polyacrylate crosspolymer-6 1%, 1,2-hexanediol 0.5%, p-hydroxyacetophenone 0.5%, arginine 0.1%, glycerin 5%, Thuja orientalis leaf pulp 80% and water.

[0067] More preferably, and as an example of the present invention, the Platycladus orientalis leaf mud film is composed of the following components by weight percentage: Polyacrylate crosspolymer-6 1%, 1,2-hexanediol 0.5%, p-hydroxyacetophenone 0.5%, arginine 0.1%, glycerin 5%, Thuja orientalis leaf pulp 80% and the balance water.

[0068] In the last aspect, the present invention provides a method for preparing the above-mentioned Platycladus orientalis leaf mud film, comprising the following steps: (1) Mix water, polyacrylate cross-linked polymer-6, 1,2-hexanediol, p-hydroxyacetophenone, arginine and glycerol uniformly, and heat to 80-85° C. to obtain phase A; (2) Heat the Platycladus orientalis leaf slurry to 80-85°C, mix it with phase A while it is still hot, and homogenize it at 4000-6000 rpm for 1-5 min to obtain Platycladus orientalis leaf mud film.

[0069] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention provides a method for preparing a Platycladus orientalis leaf extract, which adopts a circulating diafiltration extraction method and a specific concentration and membrane separation process to successfully extract three Platycladus orientalis leaf extracts, namely, Platycladus orientalis hydrosol, Platycladus orientalis leaf extract and Platycladus orientalis slurry.

[0070] (2) The Platycladus orientalis leaf extract obtained by the present invention can be effectively integrated into different cosmetic systems and has good versatility.

[0071] (3) The Platycladus orientalis leaf extract obtained by the present invention has a high content of total flavonoids and quercetin, which are effective ingredients, and can effectively regulate the water-oil balance of the skin and regulate the oil secretion of the sebaceous glands. The hair products made from the Platycladus orientalis leaf extract prepared by this method can effectively reduce the oil content of the scalp, and at the same time have the effect of reducing dandruff and oily hair. The mud mask made from the Platycladus orientalis leaf pulp obtained by this method has the effect of effectively regulating the oil secretion of the face of the subject and reducing the number of acne. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 This is a graph showing the trend of hair loss at various time points before and after using the product for effect evaluation 6.

[0073] Figure 2 This is a trend chart of the difference in the amount of hair loss at each time point after using the product and before using the product for effect evaluation 6.

[0074] Figure 3 This is a graph showing the trend of hair density changes at various time points before and after using the product for effect evaluation 6.

[0075] Figure 4 This is a trend chart of the difference in hair density at various time points after using the product and before using the product for effect evaluation 6.

[0076] Figure 5 This is a test result chart of the amount of dandruff of 6 test subjects before and after using the product.

[0077] Figure 6 This is a trend chart of the difference in the number of dandruff at each time point after using the product and before using the product for effect evaluation 6.

[0078] Figure 7 This is a test result chart of the oil content on the scalp surface of 6 test subjects before and after using the product.

[0079] Figure 8 This is a trend chart of the difference in scalp surface oil content at various time points after using the product and before using the product for effect evaluation 6.

[0080] Fig. 9 This is a graph showing the test results of the oil content on the skin surface of the test subjects in Effect Evaluation 7 before and after using the product.

[0081] Fig.10 This is a graph showing the change in the difference in skin surface oil content before and after using the product in Effect Evaluation 7.

[0082] Fig.11 This is a test result chart of the number of acne on the subjects in Effect Evaluation 7 before and after using the product.

[0083] Fig.12 This is a graph showing the change in the difference in the number of acne after using the product and before using the product in Effect Evaluation 7.

[0084] Fig.13 This is a graph showing the test results of the water content in the stratum corneum of the subjects' skin in Effect Evaluation 7 before and after using the product.

[0085] Fig.14 This is a graph showing the change in the difference in water content of the skin stratum corneum before and after using the product in Effect Evaluation 7. DETAILED DESCRIPTION

[0086] Terms and Claims of the Present Invention: 1. The articles “a”, “an” and “the” include plural referents unless expressly limited to one or more referents otherwise.

[0087] 2. Numerical range: Unless otherwise expressly stated, all ranges or ratios disclosed herein will be understood to include any and all sub-ranges or sub-ratios contained therein. For example, a range or ratio of 1 to 30 stated should be considered to be included between a minimum value of 1 and a maximum value of 30, and any sub-range or sub-ratio, integer, decimal, or sub-range or sub-ratio consisting of integers or decimals including the end points.

[0088] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0089] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all weight contents. Unless otherwise specified, it is understood that the process is carried out at room temperature.

[0090] In the following examples and comparative examples, the sources of some reagents are shown in Table 1: Table 1

[0091] Example 1 Preparation of Platycladus orientalis leaf extract: S1. Add 8000g water to 1000g Platycladus orientalis leaves (quercetin content 0.31%), circulate and filter at 90℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 6000g water, circulate and filter at 90℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 4000g water, circulate and filter at 90℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue. The residue is the medicinal residue. The combined filtrate is the circulating leachate of Platycladus orientalis leaves.

[0092] S2. The Platycladus orientalis leaf circulating leachate obtained in step S1 is concentrated to 5000 g by filtering through a 100 Dalton concentration filter membrane, and filtered until clarified to obtain Platycladus orientalis leaf concentrated filtrate.

[0093] S3, the concentrated filtrate of Platycladus orientalis leaves obtained in step S2 is concentrated to 800 g by reduced pressure distillation at 65°C, and the condensate and the concentrated solution are collected simultaneously. The condensate is sterilized to obtain Platycladus orientalis hydrosol.

[0094] S4, cooling the concentrated solution obtained in step S3 to below 30°C, adding 95% ethanol aqueous solution by volume to obtain a mixed solution. The ethanol volume fraction of the mixed solution is 20%. Cooling to 4°C, standing for 2 hours, filtering until clear, filtering through a 1000 Dalton molecular cutoff membrane to remove high molecular weight components and retain low molecular weight components. Concentrating and freeze-drying to obtain 209.8g of Platycladus orientalis leaf extract.

[0095] S5. Grind the residue obtained in step S1 into a slurry with a colloid mill to a particle size of 10-30 μm, and concentrate the slurry to a solid content of 25% to obtain Platycladus orientalis slurry.

[0096] The Platycladus orientalis slurry obtained in step S5 can be further processed to obtain Platycladus orientalis leaf powder or Platycladus orientalis leaf slurry.

[0097] S601. The preparation method of Platycladus orientalis leaf powder is as follows: freeze-drying the Platycladus orientalis slurry obtained in step S5 to obtain the Platycladus orientalis leaf powder.

[0098] S602. The method for preparing the Platycladus orientalis leaf pulp is as follows: the Platycladus orientalis slurry obtained in step S5 is concentrated to a solid weight content of 30% to obtain the Platycladus orientalis leaf pulp.

[0099] Example 2 Preparation of Platycladus orientalis leaf extract: S1. Add 16,000 g of water to 1000 g of Platycladus orientalis leaves (quercetin content 0.26%), and perform 90-minute cyclic percolation extraction at 99 ° C. The filtrate is rapidly cooled to below 60 ° C, and the residue is collected. Add 16,000 g of water, perform 90-minute cyclic percolation extraction at 99 ° C. The filtrate is rapidly cooled to below 60 ° C, and the residue is collected. The residue is the medicinal residue. The combined filtrate is the circulating percolation liquid of Platycladus orientalis leaves.

[0100] S2. The Platycladus orientalis leaf circulating leachate obtained in step S1 is concentrated to 7000 g through a 200 Dalton concentration filter membrane, and filtered until clarified to obtain a Platycladus orientalis leaf concentrated filtrate.

[0101] S3, the concentrated filtrate of Platycladus orientalis leaves obtained in step S2 is concentrated to 1400 g by reduced pressure distillation at 60°C, and the condensate and the concentrated liquid are collected simultaneously. The condensate is sterilized to obtain Platycladus orientalis hydrosol.

[0102] S4, cooling the concentrated solution obtained in step S3 to below 30°C, adding 95% ethanol aqueous solution by volume to obtain a mixed solution. The ethanol volume fraction of the mixed solution is 30%. Cooling to -6°C, standing for 2 hours, filtering until clear, filtering through a 5000 Dalton molecular cutoff membrane to remove high molecular weight components and retain low molecular weight components. Concentrating and freeze-drying to obtain 229.6g of Platycladus orientalis leaf extract.

[0103] S5. Grind the residue obtained in step S1 into a slurry with a colloid mill to a particle size of 10-30 μm, and concentrate the slurry to a solid content of 25% to obtain Platycladus orientalis slurry.

[0104] Example 3 Preparation of Platycladus orientalis leaf extract: S1. Add 10000g of water to 1000g of Platycladus orientalis leaves (quercetin content 0.31%), and extract by circulating filtration at 95℃ for 60 minutes. The filtrate is quickly cooled to below 60℃, and the residue is collected. Add 6000g of water, and extract by circulating filtration at 95℃ for 60 minutes. The filtrate is quickly cooled to below 60℃, and the residue is collected. Add 4000g of water, and extract by circulating filtration at 95℃ for 60 minutes. The filtrate is quickly cooled to below 60℃, and the residue is collected. The residue is the medicinal residue. The combined filtrate is the circulating filtrate of Platycladus orientalis leaves.

[0105] S2. The Platycladus orientalis leaf circulating leachate obtained in step S1 is concentrated to 6000 g through a 100 Dalton concentration filter membrane, and filtered until clarified to obtain a Platycladus orientalis leaf concentrated filtrate.

[0106] S3, the concentrated filtrate of Platycladus orientalis leaves obtained in step S2 is concentrated to 1000 g by vacuum distillation at 65°C, and the condensate and the concentrated solution are collected simultaneously. The condensate is sterilized to obtain Platycladus orientalis hydrosol.

[0107] S4, cooling the concentrated solution obtained in step S3 to below 30°C, adding 95% ethanol aqueous solution by volume to obtain a mixed solution. The ethanol volume fraction of the mixed solution is 25%. Cooling to 2°C, standing for 2 hours, filtering until clear, filtering through a 2000 Dalton molecular cutoff membrane to remove high molecular weight components and retain low molecular weight components. Concentrating and freeze-drying to obtain 209.1g of Platycladus orientalis leaf extract.

[0108] S5. Grind the residue obtained in step S1 into a slurry with a colloid mill to a particle size of 10-30 μm, and concentrate the slurry to a solid content of 25% to obtain Platycladus orientalis slurry.

[0109] Example 4 Preparation of Platycladus orientalis leaf extract: S1. Add 6000g water to 1000g Platycladus orientalis leaves (quercetin content 0.31%), circulate and filter at 80℃ for 90 minutes, cool the filtrate to below 60℃, collect the residue, add 4000g water, circulate and filter at 80℃ for 90 minutes, cool the filtrate to below 60℃, collect the residue, add 4000g water, circulate and filter at 80℃ for 90 minutes, cool the filtrate to below 60℃, collect the residue. The residue is the medicinal residue. The combined filtrate is the circulating leachate of Platycladus orientalis leaves.

[0110] S2. The Platycladus orientalis leaf circulating leachate obtained in step S1 is concentrated to 6000 g through a 300 Dalton concentration filter membrane, and filtered until clarified to obtain Platycladus orientalis leaf concentrated filtrate.

[0111] S3, the concentrated filtrate of Platycladus orientalis leaves obtained in step S2 is concentrated to 6000 g by reduced pressure distillation at 65°C, and the condensate and the concentrated liquid are collected simultaneously. The condensate is sterilized to obtain Platycladus orientalis hydrosol.

[0112] S4, cooling the concentrated solution obtained in step S3 to below 30°C, adding 95% ethanol aqueous solution by volume to obtain a mixed solution. The ethanol volume fraction of the mixed solution is 25%. Cooling to 0°C, standing for 3 hours, filtering until clear, filtering through a 3000 Dalton molecular cutoff membrane to remove high molecular weight components and retain low molecular weight components. Concentrating and freeze-drying to obtain 212.2 g of Platycladus orientalis leaf extract.

[0113] S5. Grind the residue obtained in step S1 into a slurry with a colloid mill to a particle size of 10-30 μm, and concentrate the slurry to a solid content of 25% to obtain Platycladus orientalis slurry.

[0114] Comparative Example 1 Preparation of Platycladus orientalis leaf extract: S1. Add 8000g of water to 1000g of Platycladus orientalis leaves (quercetin content 0.31%), circulate and filter at 99℃ for 30 minutes, filter, and collect the filtrate. Add 6000g of water to the filter residue, circulate and filter at 99℃ for 30 minutes, filter, and collect the filtrate. Add 4000g of water to the filter residue, circulate and filter at 99℃ for 30 minutes, filter, and collect the filtrate. The combined filtrate is the Platycladus orientalis leaf circulating leachate.

[0115] S2. The arborvitae leaf circulating leachate obtained in step S1 was filtered until clarified, concentrated to 800 g by reduced pressure distillation at 65°C, and the condensate and the concentrate were collected simultaneously. The condensate was sterilized to obtain arborvitae pure water. The concentrate was freeze-dried to obtain 226.8 g of arborvitae leaf extract.

[0116] Comparative Example 2 Preparation of Platycladus orientalis leaf extract: 1000g Platycladus orientalis leaves (quercetin content 0.31%) were added with 8000g ethanol aqueous solution with a volume content of 30%, and the mixture was extracted by slight boiling reflux for 30min, filtered, and the filtrate was collected. The residue was added with 6000g ethanol aqueous solution with a volume content of 30%, and the mixture was extracted by slight boiling reflux for 30min, filtered, and the filtrate was collected. The residue was added with 4000g ethanol aqueous solution with a volume content of 30%, and the mixture was extracted by slight boiling reflux for 30min, filtered, and the filtrate was collected. All the filtrates were combined, filtered until clear, and concentrated to 800g by reduced pressure distillation at 60°C, and the condensate and the concentrate were collected simultaneously. The condensate was sterilized to obtain Platycladus orientalis pure dew. The concentrate was freeze-dried to obtain 222.9g Platycladus orientalis leaf extract.

[0117] Comparative Example 3 Preparation of Platycladus orientalis leaf extract: 1000g of Platycladus orientalis leaves (quercetin content 0.31%) were added with 15000g of 55% ethanol aqueous solution, extracted at 65°C for 2h, filtered, and the filtrate was adjusted to pH 9 with sodium hydroxide solution, and filtered until clear. The clear filtrate was adjusted to pH 7 with hydrochloric acid, concentrated and spray-dried to obtain 196.5g of Platycladus orientalis leaf extract.

[0118] Comparative Example 4 Preparation of Platycladus orientalis leaf extract: 1000g of Platycladus orientalis leaves (quercetin content 0.31%) was added with 15000g of 55% ethanol aqueous solution, extracted at 65℃ for 2h, filtered, and the filtrate was adjusted to pH 9 with sodium hydroxide solution, and filtered until clear. The clear filtrate was adjusted to pH 7 with hydrochloric acid, concentrated to 3000g, 1500g of propylene glycol and 25g of PEG-40 hydrogenated castor oil were added, water was added to 5000g, stirred evenly, filtered until clear, and Platycladus orientalis leaf extract was obtained. After testing, the solid content of Platycladus orientalis leaf extract was 3.91%, and the total solid content of Platycladus orientalis leaf extract was 195.5g.

[0119] Comparative Example 5 Preparation of Platycladus orientalis leaf extract: 1000g of Platycladus orientalis leaves (quercetin content 0.31%) was added with 15000g of 55% ethanol aqueous solution, extracted at 65℃ for 2h, filtered, and the filtrate was adjusted to pH 9 with sodium hydroxide solution, and filtered until clear. The clarified filtrate was adjusted to pH 7 with hydrochloric acid, concentrated to 3000g, 1500g of propylene glycol and 5g of disodium EDTA were added, water was added to 5000g, stirred evenly, filtered until clear, and Platycladus orientalis leaf extract was obtained. After testing, the solid content of Platycladus orientalis leaf extract was 3.94%, and the total solid content of Platycladus orientalis leaf extract was 192g.

[0120] Comparative Example 6 Preparation of Platycladus orientalis leaf extract: S1. Add 8000g of water to 1000g of Platycladus orientalis leaves (quercetin content 0.11%), circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 6000g of water, circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 4000g of water, circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue. The combined filtrate is the circulating leachate of Platycladus orientalis leaves.

[0121] S2. The Platycladus orientalis leaf circulating leachate obtained in step S1 is concentrated to 5000 g by filtering through a 100 Dalton concentration filter membrane, and filtered until clarified to obtain Platycladus orientalis leaf concentrated filtrate.

[0122] S3, the concentrated filtrate of Platycladus orientalis leaves obtained in step S2 is concentrated to 800 g by reduced pressure distillation at 65°C, and the condensate and the concentrated solution are collected simultaneously. The condensate is sterilized to obtain Platycladus orientalis hydrosol.

[0123] S4, cooling the concentrated solution obtained in step S3 to below 30°C, adding 95% ethanol aqueous solution by volume to obtain a mixed solution. The ethanol volume fraction of the mixed solution is 20%. Cooling to 4°C, standing for 2 hours, filtering until clear, filtering through a 1000 Dalton molecular cutoff membrane to remove high molecular weight components and retain low molecular weight components. Concentrating and freeze-drying to obtain 208.5 g of Platycladus orientalis leaf extract.

[0124] Comparative Example 7 Preparation of Platycladus orientalis leaf extract: S1. Add 8000g of water to 1000g of Platycladus orientalis leaves (quercetin content 0.11%), circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 6000g of water, circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 4000g of water, circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue. The combined filtrate is the circulating leachate of Platycladus orientalis leaves.

[0125] S2. The Platycladus orientalis leaf circulating leachate obtained in step S1 is concentrated to 5000 g by filtering through a 100 Dalton concentration filter membrane, and filtered until clarified to obtain Platycladus orientalis leaf concentrated filtrate.

[0126] S3, the concentrated filtrate of Platycladus orientalis leaves obtained in step S2 is concentrated to 800 g by reduced pressure distillation at 65°C, and the condensate and the concentrated solution are collected simultaneously. The condensate is sterilized to obtain Platycladus orientalis hydrosol.

[0127] S4. Cool the concentrated solution obtained in step S3 to below 30°C, add 95% ethanol aqueous solution by volume to obtain a mixed solution. The ethanol volume fraction of the mixed solution is 20%. Cool to 4°C, let stand for 2 hours, filter until clear, concentrate, and freeze-dry to obtain 214g of Platycladus orientalis leaf extract.

[0128] Comparative Example 8 Preparation of Platycladus orientalis leaf extract: S1. Add 8000g of water to 1000g of Platycladus orientalis leaves (quercetin content 0.11%), circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 6000g of water, circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue, add 4000g of water, circulate and filter at 99℃ for 30 minutes, cool the filtrate to below 60℃, collect the residue. The combined filtrate is the circulating leachate of Platycladus orientalis leaves.

[0129] S2. The Platycladus orientalis leaf circulating leachate obtained in step S1 is concentrated to 5000 g by filtering through a 100 Dalton concentration filter membrane, and filtered until clarified to obtain Platycladus orientalis leaf concentrated filtrate.

[0130] S3, the concentrated filtrate of Platycladus orientalis leaves obtained in step S2 is concentrated to 800 g by reduced pressure distillation at 65°C, and the condensate and the concentrated solution are collected simultaneously. The condensate is sterilized to obtain Platycladus orientalis hydrosol.

[0131] S4, cooling the concentrated solution obtained in step S3 to below 30°C, adding 95% ethanol aqueous solution by volume to obtain a mixed solution. The ethanol volume fraction of the mixed solution is 20%. Cooling to 4°C, standing for 2 hours, filtering until clear, filtering through a 10,000 Dalton molecular cutoff membrane to remove high molecular weight components and retain low molecular weight components. Concentrating and freeze-drying to obtain 212.8 g of Platycladus orientalis leaf extract.

[0132] Effect evaluation 1 The total flavonoids content and quercetin content in the Platycladus orientalis leaf extracts prepared in Examples 1 to 4 and Comparative Examples 1 to 8 were detected.

[0133] Determination method: 1. Test of water-soluble total flavonoids content 1.1 Equipment and Materials UV spectrophotometer (TU-1900), Nessler colorimetric tube, quartz cuvette, electronic balance, rutin, sodium nitrite, aluminum nitrate, sodium hydroxide.

[0134] 1.2 Experimental Methods 1.2.1. Preparation of Rutin Standard Solution Accurately weigh 10.0 mg of rutin standard that has been dried under reduced pressure to constant weight, dilute to a 25 mL volumetric flask with methanol, use ultrasonic dissolution, and cool to room temperature before diluting to volume; or take 20 mg of rutin standard and dilute to a 50 mL volumetric flask with methanol, depending on the amount used.

[0135] 1.2.2 Preparation of sodium nitrite solution (5%) Accurately weigh 5.0 g of sodium nitrite and dilute to 100 mL with deionized water in a volumetric flask.

[0136] 1.2.3. Preparation of aluminum nitrate solution (10%) Accurately weigh 17.601 g of aluminum nitrate nonahydrate (Al(NO 3 ) 3 9H 2 O), dissolve in deionized water and dilute to 100 mL using a volumetric flask.

[0137] 1.2.4 Preparation of sodium hydroxide solution (4%) Accurately weigh 10.0 g of sodium hydroxide, dissolve it in deionized water, and after cooling, use a volumetric flask to make up to 250 mL.

[0138] 1.2.5 Preparation of test solution The sample was diluted with deionized water from the original concentration to a flavonoid concentration in the range of 0.4-2.0 mg / mL and filtered until clear.

[0139] 1.2.6. Drawing of standard curve Take 6 25mL colorimetric tubes numbered 0#-5#, and accurately pipette 0mL, 1.0mL, 2.0mL, 3.0mL, 4.0mL, and 5.0mL of rutin standard solution into each group 0#-5#, and fill each tube to 6.0mL with deionized water and mix well. Add 1.0mL of 5% sodium nitrite solution to each group, mix well, and place accurately at room temperature for 6min, then add 1.0mL of 10% aluminum nitrate solution to each group, mix well, and place accurately at room temperature for 6min, and finally add 10.0mL of 4% sodium hydroxide solution to each group, fill water to the 25mL scale line, mix well, and place accurately at room temperature for 15min. Take the solution numbered 0# as the blank, measure the absorbance of the reaction solution of each group 1#-5# at a wavelength of 510nm, and draw a standard curve with the concentration of rutin standard solution as the horizontal axis and the absorbance as the vertical axis.

[0140] 1.2.7 Determination Pipette 1.0mL of the test solution, add water to 6mL, add 1.0mL of 5% sodium nitrite solution, mix well, react at room temperature for 6min, add 1.0mL of 10% aluminum nitrate solution, mix well, react at room temperature for 6min, and finally add 10.0mL of 4% sodium hydroxide solution, add water to the 25mL mark, mix well, react at room temperature for 15min. Measure the absorbance of the reaction solution at a wavelength of 510nm. Use the solution numbered 0# as the blank, and make 2 parallels for each group of test solutions.

[0141] 1.2.8 Calculation Calculate the result according to the following formula: X=c×N.

[0142] Where: X is the content of flavonoids in the sample, in milligrams per milliliter (mg / g); c is the mass concentration of flavonoids in the test solution measured by the standard curve, in milligrams per milliliter (mg / mL); c=(Ab)÷a, A is the absorbance of the sample, and a and b are the parameters of the linear equation Y=aX+b.

[0143] N is the dilution factor of the test solution.

[0144] 2. Quercetin content test 2.1 Equipment and Materials High performance liquid chromatography (Shimadzu LC-16), chromatographic column: ZORBAXSB-C18 (4.6×250mm, 5μm), electronic balance, quercetin (analytical standard), methanol (analytical grade), and ultrapure water.

[0145] 2.2 Experimental Methods 2.2.1 Chromatographic conditions (a) Chromatographic column: ZORBAXSB-C18 (4.6×250 mm, 5 μm); (b) Flow rate: 1.0 mL / min; (c) Mobile phase: methanol / 0.1% phosphoric acid aqueous solution = 40 / 60, (40 min); (d) Detection wavelength: 350 nm; (e) Column temperature: 30°C; (f) Injection volume: 10 µL.

[0146] 2.2.2 Quercetin Standard Weigh 10.0 mg (accurate to 0.1 mg), dissolve in methanol to 10 mL to obtain mother solution, take 1 mL of mother solution, dilute with 50% methanol to 10 mL to obtain quercetin standard. Take 5µL, 10µL, 15µL, 20µL, and 25µL of quercetin standard for analysis, and draw a standard curve with injection volume (µg / mL) as the ordinate and peak area as the abscissa. Alternatively, prepare a series of concentration samples with an injection volume of 10µL. Draw a standard curve with peak area as the abscissa and injection volume (µg / mL) as the ordinate.

[0147] 2.2.3 Sample determination Take the sample and dilute it with 50% methanol to 10mL, pass it through a 0.45µm membrane and set aside. Take 10μL of the prepared sample and inject it under the above chromatographic conditions for analysis, and perform qualitative and quantitative analysis based on the retention time (min).

[0148] 2.2.4 Calculation Calculate the result according to the following formula

[0149] Where: S is the quercetin content, in micrograms per milliliter (μg / mL); C 0 is the concentration of the standard, in milligrams per milliliter (μg / mL); A 0 is the peak area of ​​the standard; A 1 is the peak area of ​​the test product; N is the dilution factor of the test sample. N =1.

[0150] The experimental results are shown in Table 2.

[0151] Table 2

[0152] Among them, the yield of Platycladus orientalis leaf extract = the weight of Platycladus orientalis leaf extract ÷ the weight of Platycladus orientalis leaf medicinal materials input; the yield of total water-soluble flavonoids = the total amount of water-soluble total flavonoids ÷ the weight of Platycladus orientalis leaf medicinal materials input; the total amount of water-soluble total flavonoids = the weight of Platycladus orientalis leaf extract × the content of water-soluble total flavonoids; the yield of quercetin = the total amount of quercetin ÷ the weight of Platycladus orientalis leaf medicinal materials input; the total amount of quercetin = the weight of Platycladus orientalis leaf extract × the content of quercetin.

[0153] For Comparative Examples 4 and 5, the total amount of water-soluble total flavonoids = the weight of Platycladus orientalis leaf extract × the content of water-soluble total flavonoids; the total amount of quercetin = the weight of Platycladus orientalis leaf extract × the content of quercetin.

[0154] From the above detection and calculation results, there is no obvious difference in the extract yield of the extracts in the embodiments and comparative examples, indicating that the extraction processes and Platycladus orientalis leaves with different quercetin contents have little effect on the extraction yield of Platycladus orientalis leaf extract.

[0155] The extraction yield of the total water-soluble flavonoids in Example 1, Example 3 and Example 4 is higher than that in Comparative Example 2 and Comparative Example 3, indicating that for the extraction of Platycladus orientalis leaves with the same quercetin content, the extraction yield of the total water-soluble flavonoids provided by the method of the present invention is significantly higher than that of the extraction process mainly based on alcohol extraction; from the extraction yields of Example 1 and Comparative Example 6 and the extraction yields of water-soluble flavonoids, the extraction yield of the Platycladus orientalis leaves with a high quercetin content will also be high, but it does not show the same multiple relationship. The water-soluble flavonoid content tested in Comparative Example 4 is significantly higher than that in Comparative Example 3 and Comparative Example 5, indicating that PEG-40 hydrogenated castor oil can solubilize water-insoluble flavonoids in water.

[0156] The quercetin extraction yields of Examples 1 to 4 are significantly higher than those of Comparative Examples 1 to 5, indicating that the extraction yield of quercetin by the technical process of the present invention is significantly higher than that of traditional water extraction, concentration, spray drying and alcohol extraction, concentration, spray drying extraction technologies; wherein the quercetin content of Comparative Examples 1 and 2 is significantly lower than that of Example 1, indicating that long-term high temperature reduces the quercetin yield; the quercetin content of Comparative Examples 3 to 5 is lower than that of Example 1, indicating that steps such as alkali treatment will reduce the quercetin yield.

[0157] Compared with Example 1, the extraction yields of Platycladus orientalis leaves, water-soluble flavonoids and quercetin in Comparative Examples 7 and 8 are similar, indicating that the filtration step of removing macromolecular substances has little effect on the extraction yield.

[0158] Effect evaluation 2 Effects of Platycladus orientalis leaf extract on keratinocyte survival rate and barrier factors.

[0159] 1. Survival rate experiment The Platycladus orientalis leaf extracts obtained in Examples 1 to 4 and Comparative Examples 1 to 8 were dissolved in deionized water and filtered to prepare samples. Each group of samples and their corresponding extract amounts and crude drug amounts are shown in Table 3.

[0160] Table 3

[0161] Under sterile conditions, the samples to be tested were diluted with culture medium, and each sample to be tested was uniformly diluted to the crude drug amount of Platycladus orientalis leaves: 0.01%, 0.1%, and 1%.

[0162] HaCaT cells were cultured at 5 × 10 3 The cells were inoculated in a 96-well plate at a density of 100 cells / well. To avoid errors caused by evaporation of the culture medium due to edge effect, only PBS was added around the 96-well plate without inoculating cells. The actual number of wells inoculated with cells was 60. The cells were incubated at 37°C and 5% CO 2 Culture in a cell culture incubator for 24 h. When the cell density is about 50%, treat the sample to be tested, and make 3 replicates for each group. After 24 h of adding the sample, the cell viability is detected by CCK-8 method, and the cell viability is calculated according to the following formula.

[0163] 00%.

[0164] Note: Background OD value is not reflected in the data.

[0165] The experimental results are shown in Table 4 and Table 5. Since the number of holes in the orifice plate is limited, Table 4 and Table 5 are the experimental results of two different orifice plates.

[0166] Table 4

[0167] Table 5

[0168] From the results in the above table, it can be seen that the Platycladus orientalis leaf extract of the embodiment of the present invention has an improving effect on the activity of keratinocytes at 0.01%, 0.1%, and 1% Platycladus orientalis leaf crude drug content; Comparative Examples 1-3 have no effect on the activity of keratinocytes at 0.01% and 0.1% Platycladus orientalis leaf crude drug content, but have a significant inhibitory effect on the activity of keratinocytes at 1% Platycladus orientalis crude drug content, reflecting a certain cytotoxicity, indicating that the process of the present invention has the effect of reducing cytotoxic substances compared with traditional water extraction, alcohol extraction, alkali-alcohol treatment and other processes; Comparative Examples 4 and Comparative Examples 5 significantly inhibit the activity of keratinocytes at 0.1% crude drug content, especially Comparative Example 4 has obvious inhibition, reflecting a strong cytotoxicity. The results show that the solubilizing agent, polyol solvent and the like have a significant inhibitory effect on the activity of keratinocytes; compared with Example 1, the promoting effect of Comparative Example 6 on cell activity is slightly lower, and the effect is not significant; Comparative Examples 7 and 8 have no effect on the activity of keratinocytes at 0.01% of the crude drug content of Platycladus orientalis leaves, and have a promoting effect on the activity of keratinocytes at 0.1% of the crude drug content of Platycladus orientalis leaves, but the effect is significantly weaker than that of Example 1, and have a certain inhibitory effect on the activity of keratinocytes at 1% of the crude drug content of Platycladus orientalis leaves, indicating that the removal of macromolecular substances is beneficial to the Platycladus orientalis leaf extract to improve the activity of keratinocytes and reduce the effect of the components that inhibit the activity of keratinocytes, and the process effect of the removed macromolecular substances above 10,000 Daltons is not obvious.

[0169] 2. Cell barrier factor experiment.

[0170] Under sterile conditions, the samples to be tested were diluted with culture medium, and each sample to be tested was uniformly diluted to the crude drug amount of Platycladus orientalis leaves: 0.025%, 0.05%, and 0.1%.

[0171] HaCaT cells were cultured in DMEM complete medium at 37°C and 5% CO 2 The cells were cultured in an incubator and subcultured when the cells reached 80% fusion. The cells were digested with trypsin to remove them from the culture flask and inoculated into a 6-well plate with approximately 2 × 10 cells per well. 5 After culturing for 24 hours, several samples of different concentrations were added and cultured for 24 hours. Total RNA in the cells was extracted, and after removing gDNA, it was reverse transcribed into cDNA for qPCR experiments. A 96-well plate was used as a 20 μL system, and the reference gene and target gene of each component were made into 3 replicates. The relative expression of the gene was calculated according to the following formula: Relative gene expression = 2 -ΔΔCt .

[0172] ΔCt = Ct value of target point in each group - average Ct value of corresponding internal reference in each group.

[0173] ΔΔCt=ΔCt value of target point in sample group-average ΔCt value of target point in blank group.

[0174] The experimental results are shown in Tables 6 and 7. Since the number of wells in each well plate is limited, Tables 6 and 7 are the results of the effects of the samples on the expression of FLG at the mRNA level in HaCaT cells in two different well plates.

[0175] Table 6

[0176] Table 7

[0177] Tables 8 and 9 show the effects of the samples on the expression of Caspase 14 at the mRNA level in HaCaT cells on two different well plates.

[0178] Table 8

[0179] Table 9

[0180] It can be seen from Tables 6 and 7 that the Platycladus orientalis leaf extracts of each embodiment have a good promoting effect on the gene expression of FLG in HaCaT cells, and at a content of 0.1% or less of Platycladus orientalis leaf crude drug, with the increase of concentration, the promoting effect also increases significantly, showing a certain dose-effect relationship; the Platycladus orientalis leaf extract of Comparative Example 1 has an effect equivalent to that of the embodiment, while the Platycladus orientalis leaf extract of Comparative Example 2 has a certain promoting effect on the gene expression of FLG in HaCaT cells, but compared with the embodiment, its effect is significantly lower, indicating that the effect of alcohol extraction is not as good as that of water extraction; the Platycladus orientalis leaf extract of Comparative Example 3, When the content of Platycladus orientalis crude drug is 0.05%, it has a certain promoting effect on the gene expression of FLG in HaCaT cells, but there is no promoting effect when the concentration reaches 0.1%, indicating that the effect of Platycladus orientalis leaf extract treated with alcohol extraction and alkali on promoting the gene expression of FLG in HaCaT cells is worse than that in the embodiment; the Platycladus orientalis leaf extracts of Comparative Examples 6 to 8 are equivalent to those of the embodiment, indicating that the content of quercetin in Platycladus orientalis medicinal materials and the content of Platycladus orientalis crude drug at 0.1% or less in the macromolecule filtration step have basically no effect on the effect of Platycladus orientalis leaf extract on promoting the gene expression of FLG in HaCaT cells.

[0181] It can be seen from Tables 8 and 9 that the Platycladus orientalis leaf extracts of each embodiment have a good promoting effect on the gene expression of Caspase14 in HaCaT cells, and when the content of Platycladus orientalis leaf crude drug is 0.1% or less, with the increase of concentration, the promoting effect also increases significantly, showing a certain dose-effect relationship; although the Platycladus orientalis leaf extract of Comparative Example 1 has a certain promoting effect on the gene expression of Caspase14 in HaCaT cells, compared with the embodiment, its effect is significantly lower, indicating that the Platycladus orientalis leaf extract extracted by this process is significantly superior to the traditional water extraction process in promoting the gene expression of Caspase14 in HaCaT cells; the Platycladus orientalis leaf extracts of Comparative Examples 2 and 3 have almost no effect on the gene expression of Caspase14 in HaCaT cells, indicating that the alcohol-extracted Platycladus orientalis leaf extract has no promoting effect on the gene expression of Caspase14 in HaCaT cells; the Platycladus orientalis leaf extract of Comparative Example 6 has an effect equivalent to that of the embodiment, indicating that the content of quercetin in Platycladus orientalis leaf medicinal materials has basically no effect on the effect of Platycladus orientalis leaf extract on promoting the gene expression of Caspase14 in HaCaT cells. Comparative Examples 7-8: When the Platycladus orientalis leaf extract contains 0.05% or less Platycladus orientalis leaf crude drug content, the effect is equivalent to that of the embodiment, but at 0.1%, the promoting effect is improved, but compared with Example 1, the promoting effect is obviously weaker than that of Example 1, indicating that the removal of macromolecular substances in Platycladus orientalis leaf medicinal materials can significantly improve the effect of Platycladus orientalis leaf extract on promoting the gene expression of Caspase14 in HaCaT cells under high concentration conditions.

[0182] Effect evaluation 3 Effect of Platycladus orientalis leaf extract on sebum synthesis in sebaceous gland cells.

[0183] Cytotoxicity test experimental method: dilute the sample to be tested with culture medium under sterile conditions, and dilute each sample to a uniform amount of Platycladus orientalis leaf crude drug: 0.01%, 0.05%, and 0.1%.

[0184] SZ95 cells were cultured at 1×10 4 The cells were inoculated in a 96-well plate at a density of 100 cells / well. To avoid errors caused by evaporation of the culture medium due to edge effect, only PBS was added around the 96-well plate without inoculating cells. The actual number of wells inoculated with cells was 60. The cells were incubated at 37°C and 5% CO 2 Culture in a cell culture incubator for 24 h. When the cell density is about 50%, treat the sample to be tested, and make 3 replicates for each group. After 24 h of adding the sample, the cell viability is detected by CCK-8 method, and the cell viability is calculated according to the following formula.

[0185] .

[0186] Note: Background OD value is not reflected in the data.

[0187] Experimental method for testing sebum content: dilute the sample to be tested with culture medium under sterile conditions, and uniformly dilute each sample to the amount of Platycladus orientalis leaf crude drug: 0.01%, 0.05%, and 0.1%.

[0188] Take the SZ95 cell suspension in the logarithmic phase and growing well and inoculate it in a black 96-well plate. To avoid errors caused by evaporation of the culture medium due to edge effect, only PBS is added around the 96-well plate without inoculating cells. The actual number of inoculated cell wells is 60, and the number of cells in each well is 1×10 4 After 24 h of incubation, samples of different concentrations were added, and three replicates were made for each group. After 24 h of incubation, the culture medium was discarded, the cells were washed twice with PBS, and 10 μg / mL Nile Red dye was added for staining. The cells were incubated at 37°C and 5% CO 2 Incubate in a cell culture incubator in the dark for 30 min, and use a microplate multifunctional detector to set the wavelength to EX / EM = 485 / 565 nm to detect the fluorescence intensity. The relative lipid content is calculated according to the following formula:

[0189] Note: Background fluorescence intensity values ​​are not reflected in the data.

[0190] The experimental results are shown in Table 10 and Table 11. Table 10 and Table 11 are respectively the results of testing the toxicity of Platycladus orientalis leaf extract on sebaceous gland cells by CCK8 method on two different well plates.

[0191] Table 10

[0192] Table 11

[0193] According to the above experimental results, when the content of the crude drug of Platycladus orientalis leaves is 0.1% or less, the Platycladus orientalis leaf extracts of each embodiment and comparative example have no toxicity to sebaceous gland cells. At this concentration, the effect of Platycladus orientalis leaf extract on the synthesis of sebum in sebaceous gland cells was tested, and the results are shown in Table 12 and Table 13. Table 12 and Table 13 are the results of the effect of the samples on the sebum content in sebaceous gland cells on two different well plates.

[0194] Table 12

[0195] Table 13

[0196] It can be seen from Tables 12 and 13 that the Platycladus orientalis leaf extracts of each embodiment have a good inhibitory effect on the synthesis of sebum in sebaceous gland cells, and with the increase of concentration, the inhibitory effect also increases, showing a certain dose-effect relationship; the Platycladus orientalis leaf extracts of Comparative Examples 1 and 2 also have a certain effect on the synthesis of sebum in sebaceous gland cells, but their effects are significantly weaker than those of the Platycladus orientalis leaf extracts of the embodiment, indicating that the Platycladus orientalis leaf extracts prepared by this process are superior to the Platycladus orientalis leaf extracts prepared by spray drying after traditional water extraction or alcohol extraction in terms of oil control effect; Comparative Examples 3 to 5 also have certain effects, but the effects are The effect of the Platycladus orientalis leaf extract is worse than that of the above-mentioned embodiments and comparative examples, indicating that alkali treatment further reduces its effect of inhibiting sebum synthesis in sebaceous gland cells; Comparative Example 6 has a certain inhibitory effect on the synthesis of sebum in sebaceous gland cells, and its effect is significantly reduced compared with Example 1, indicating that the effect of the Platycladus orientalis leaf extract prepared from Platycladus orientalis leaves with low quercetin content in inhibiting sebum synthesis in sebaceous gland cells is not as good as that prepared from Platycladus orientalis leaves with high quercetin content; Comparative Examples 7 and 8 are equivalent to Example 1, indicating that removing large molecules does not affect the effect of Platycladus orientalis leaf extract in inhibiting sebum synthesis in sebaceous gland cells.

[0197] Effect evaluation 4 The effect of arborvitae hydrosol on skin lipid balance.

[0198] The Platycladus orientalis hydrosol prepared in Examples 1 to 4 and Comparative Example 1 was used as a research object to study its effect on the balance of skin lipids.

[0199] Experimental method: Dilute the samples to be tested with culture medium under sterile conditions, and dilute each sample to 0.2%, 1%, and 5%.

[0200] Effect on cell survival rate: HaCaT or SZ95 cells were cultured at 5×10 3 or 1×10 4 The cells were inoculated in 96-well plates at a density of 100 cells / well. To avoid errors caused by evaporation of the culture medium due to edge effects, only PBS was added around the 96-well plate without inoculating cells. The actual number of wells inoculated with cells was 60. The cells were incubated at 37 °C and 5% CO. 2 Culture in a cell culture incubator for 24 h. When the cell density is about 50%, treat the sample to be tested, and make 3 replicates for each group. After 24 h of adding the sample, the cell viability was detected using the CCK-8 method, and the cell viability was calculated according to the following formula.

[0201] .

[0202] Note: Background OD value is not reflected in the data.

[0203] Effects on the expression of barrier factors in keratinocytes: HaCaT cells were cultured in DMEM complete medium at 37°C and 5% CO 2 The cells were cultured in an incubator and subcultured when the cells reached 80% fusion. The cells were digested with trypsin to remove them from the culture flask and inoculated into a 6-well plate with approximately 2 × 10 cells per well. 5 After culturing for 24 hours, several samples of different concentrations were added and cultured for 24 hours. Total RNA in the cells was extracted, and after removing gDNA, it was reverse transcribed into cDNA for qPCR experiments. A 96-well plate was used as a 20 μL system, and the reference gene and target gene of each component were made into 3 replicates. The relative expression of the gene was calculated according to the following formula: Relative gene expression = 2 -ΔΔCt ΔCt = Ct value of target point in each group - average Ct value of corresponding internal reference in each group ΔΔCt = ΔCt value of target point in sample group - average ΔCt value of target point in blank group Effect on sebum synthesis by sebaceous gland cells: SZ95 cell suspension in the logarithmic phase and growing well was inoculated in a black 96-well plate. To avoid errors caused by evaporation of culture medium due to edge effect, only PBS was added around the 96-well plate without inoculating cells. The actual number of inoculated cell wells was 60, with 1×10 cells per well. 4 After 24 h of incubation, samples of different concentrations were added, and three replicates were made for each group. After 24 h of incubation, the culture medium was discarded, the cells were washed twice with PBS, and 10 μg / mL NileRed dye was added for staining. The cells were incubated at 37°C and 5% CO 2 Incubate in a cell culture incubator in the dark for 30 min, and use a microplate multifunctional detector to set the wavelength to EX / EM = 485 / 565 nm to detect the fluorescence intensity. The relative lipid content is calculated according to the following formula: .

[0204] Note: Background fluorescence intensity values ​​are not reflected in the data.

[0205] The experimental results are shown in Tables 14 and 15. Tables 14 and 15 respectively show the effects of Platycladus orientalis hydrosol on the cell survival rate of SZ95 sebaceous gland cells on two different well plates.

[0206] Table 14

[0207] Table 15

[0208] Table 16 shows the effect of Platycladus orientalis hydrosol on sebum synthesis in SZ95 sebaceous gland cells.

[0209] Table 16

[0210] Tables 17 and 18 respectively show the effects of Platycladus orientalis hydrosol on the gene expression of very long-chain fatty acid elongase ELOVL4 and ELOVL7 in keratinocytes.

[0211] Table 17

[0212] Table 18

[0213] Table 19 shows the effect of Platycladus orientalis hydrosol on the gene expression of ceramide synthase CerS4 in keratinocytes.

[0214] Table 19

[0215] It can be seen from Tables 14 and 15 that at concentrations of 5% or less, the Platycladus orientalis hydrosols of each example and the Platycladus orientalis hydrosol of Comparative Example 1 have no inhibitory effect on sebaceous gland cells and keratinocytes, and cell molecular biology research can be carried out at these concentrations.

[0216] It can be seen from Table 16 that the Platycladus orientalis hydrosol of each embodiment has a good effect of inhibiting the synthesis of oil by sebaceous gland cells at concentrations of 0.02%, 1%, and 5%, and shows a good dose-effect relationship; while the effect of the Platycladus orientalis hydrosol of Comparative Example 1 is far different.

[0217] It can be seen from Table 17 that the Platycladus orientalis hydrosol of each example has a significant promoting effect on the expression of the very long-chain fatty acid elongase ELOVL4 gene in keratinocytes at concentrations of 1% and 5%, especially at a concentration of 5%. However, the Platycladus orientalis hydrosol of comparative example 1 has only a weak promoting effect on the expression of the gene at concentrations of 1% and 5%, and its promoting effect is not significantly improved when the concentration is increased from 1% to 5%.

[0218] It can be seen from Table 18 that the Platycladus orientalis hydrosol of each example has a significant promoting effect on the expression of the very long-chain fatty acid elongase ELOVL7 gene in keratinocytes at concentrations of 1% and 5%, and the promoting effect is significantly improved with the increase of concentration; while the Platycladus orientalis hydrosol of comparative example 1 only shows a significant promoting effect at a concentration of 5%, and is far inferior to the promoting effect of the Platycladus orientalis hydrosol of the example.

[0219] It can be seen from Table 19 that the Platycladus orientalis hydrosol of each example has a significant promoting effect on the expression of ceramide synthase CerS4 gene in keratinocytes at a concentration of 5%, while the Platycladus orientalis hydrosol of comparative example 1 does not show a significant promoting effect at a concentration of 5%.

[0220] In summary, the Platycladus orientalis hydrosol of the present invention has a significant promoting effect on the enhancement of intercellular lipids and a significant inhibiting effect on sebaceous gland oil synthesis.

[0221] Example 5 A shampoo containing Platycladus orientalis leaf extract. The formula and preparation process are shown in Table 20.

[0222] Table 20

[0223] This anti-hair loss shampoo is a transparent anti-hair loss shampoo. When the Platycladus orientalis leaf extract in phase B is the Platycladus orientalis leaf extract provided in Examples 1 to 4 and Comparative Examples 1 to 8, the corresponding shampoo groups are recorded as Shampoo Example 1 to Shampoo Example 4 and Shampoo Comparative Example 1 to Shampoo Comparative Example 8, respectively.

[0224] Phase C of the above formula contains three ingredients: polyquaternium-10, glycerol, and water. The content of polyquaternium-10 can be adjusted to 0.2% or 0.3%, and at this time, the content of water in phase C is adjusted to 10.2% or 10.1%, respectively.

[0225] Example 6 A scalp essence containing Platycladus orientalis leaf extract, the formula and preparation process of which are shown in Table 21.

[0226] Table 21

[0227] Among them, when the Platycladus orientalis leaf extract in phase B is the Platycladus orientalis leaf extract provided in Examples 1 to 4 and Comparative Examples 1 to 8, respectively, the corresponding scalp essence groups are recorded as Scalp Essence Example 1 to Scalp Essence Example 4 and Scalp Essence Comparative Example 1 to Scalp Essence Comparative Example 8, respectively; the content of butylene glycol in phase B can also be replaced by 0%, 5%, 10% or 20%.

[0228] Effect evaluation 5 Compatibility evaluation of Platycladus orientalis leaf extract in different head care daily chemical products.

[0229] 1. Compatibility of Platycladus orientalis leaf extract in shampoo.

[0230] The shampoos of each group provided in Example 5 were evaluated for visual clarity and centrifugal stability (5000 rpm, 30 min) after being placed at 4°C for one week, based on the amount of cationic conditioning agent polyquaternium-10. The evaluation results are shown in Table 22.

[0231] Table 22

[0232] In the transparent system shampoo provided in Example 5, as a transparent system, the cationic conditioning agent is a key component to improve the smoothness of wet combing and reduce the tangling of wet combing hair, and the compatibility of Platycladus orientalis leaf extract with cations is crucial. As can be seen from the results in the table, the Platycladus orientalis leaf extract of each embodiment and Comparative Example 6 is used in a transparent anti-hair loss shampoo system, and has good compatibility, and can be well compatible with 0.2%, 0.3%, and 0.4% added amounts of polyquaternium-10; while the compatibility of Comparative Examples 1-3, 7, and 8 is relatively poor, and cannot be used in combination with 0.2%, 0.3%, and 0.4% added amounts of polyquaternium-10. Among them, Comparative Example 6 differs from Example 1 only in the content of raw material Platycladus orientalis leaf quercetin, and the other processes are consistent, indicating that the process of the present invention removes components incompatible with cations during the extraction and refining process of Platycladus orientalis leaves.

[0233] 2. The compatibility of Platycladus orientalis leaf extract in scalp essence.

[0234] The scalp essences of each group provided in Example 6 were evaluated for visual clarity and centrifugal stability (5000 rpm, 30 min) after being placed at 4°C for 1 week, with the dosage of butylene glycol adjusted to 0%, 5%, 10%, and 20%. The evaluation results are shown in Table 23.

[0235] Table 23

[0236] The scalp essence provided in Example 6 is an aqueous system containing a certain amount of polyols. The compatibility of the Platycladus orientalis leaf extract with this type of formula depends on the stability of the Platycladus orientalis leaf in water or a formula containing a certain amount of polyols.

[0237] As can be seen from the above table, the Platycladus orientalis leaf extracts of each embodiment and comparative example 6 all show good compatibility in the essence systems of 0%, 5%, 10%, and 20% polyols; while the compatibility of the Platycladus orientalis leaf extracts prepared by comparative examples 1 to 3, 7, and 8 is relatively poor, showing problems such as turbidity and precipitation. Among them, comparative example 6 and embodiment 1 differ only in the content of the raw material Platycladus orientalis leaf quercetin, and the other processes are consistent. This shows that the Platycladus orientalis leaf extract prepared by the process of the present invention has removed water-insoluble impurities, impurities that are not tolerated in low-concentration alcohol solutions, and impurities that cause precipitation during low-temperature storage, thereby greatly improving the compatibility of the Platycladus orientalis leaf extract.

[0238] Effect evaluation 6 Consumer reviews of Thuja orientalis shampoo and scalp serum.

[0239] 1. Materials and instruments 1.1 Sample For the evaluation of Platycladus orientalis leaf shampoo, the test sample group used the shampoo Example 1 with 0.4% polyquaternium-10 added, and the control group used the shampoo without adding Platycladus orientalis leaf extract. The difference between the control group shampoo and the shampoo Example 1 was that during the preparation process, the Platycladus orientalis leaf extract was replaced with an equal weight of deionized water.

[0240] For the evaluation of the scalp essence, the test sample group used the scalp essence Example 1 with 2% butylene glycol added, and the control group used the scalp essence without adding the Platycladus orientalis leaf extract. The scalp essence of the control group was different from the scalp essence Example 1 only in that, during the preparation process, the Platycladus orientalis leaf extract was replaced with an equal weight of deionized water.

[0241] 1.2. Usage Staff members will apply the test sample (shampoo or scalp essence) to the subjects three times a week, using it on half of their head, and the corresponding control group will be used on the other side as a control.

[0242] How to use shampoo: wet your hair with clean water, take an appropriate amount of shampoo and rub it for 1-2 minutes, then rinse off the foam.

[0243] To use the scalp essence, apply it to the scalp, apply it evenly in clockwise circles with your fingertips, and finally blow-dry your hair or let it dry naturally.

[0244] 1.3 Instruments and Software Scalp analyzer (KC, South Korea, QM-SCALP-002), multi-functional skin tester MPA580 (CK, Germany), ImageJ1.52a (NIH, USA).

[0245] 1.4 Test indicators 1.4.1. Oil content on the scalp surface The scalp oil content test is completed by the SM815 probe of the MPA 580 skin tester of the German CK company. The probe contains a special matte tape with a width of 8 mm and a thickness of 0.1 mm. When this tape comes into contact with the oil on the skin, it will gradually become translucent and the light transmittance will increase. The amount of light transmittance directly reflects the oil content: the more oil, the greater the light transmittance. The test result range is 0-350μg / cm 2 The higher the value, the more oil there is on the scalp.

[0246] 1.4.2. Dandruff quantity The subjects were prohibited from washing their hair by themselves during the study period, and were not allowed to comb their hair on the morning of the visit. After arriving at the laboratory, the same technician used a comb of uniform specifications to comb the hair 30 times on each side in the order from left front to left back and right front to right back to collect dandruff on a black plastic shell, and then took pictures of the collected dandruff with a SLR camera, and used ImageJ to count the number of dandruff in the photos. The number of dandruff is measured in "grains", and the larger the value, the more serious the dandruff problem.

[0247] 1.4.3. Number of hair loss The subjects were not allowed to wash their hair during the study period and were not allowed to comb their hair on the morning of the visit. After arriving at the laboratory, the same technician used a comb of uniform specifications to comb the hair 30 times on each side in the order of left front to left back and right front to right back. Afterwards, the hair that fell off on both sides was collected and counted.

[0248] 1.4.4 Hair density The hair in the test area needs to be cut to less than 1 mm. Use the scalp analyzer of South Korea's KC company to image the scalp and hair in the test area and count the number of hairs in the photo. At the same time, measure the area of ​​the scalp area according to the scale in the photo. And calculate the hair density according to the following formula. The higher the hair density D value, the denser the hair per unit area: D=N / S Note: N is the number of hair roots, S is the scalp area (cm 2 ), D represents the hair density (roots / cm 2 ).

[0249] 2. Test methods 2.1. Subject requirements: According to the Declaration of Helsinki, the selection of subjects follows the medical and ethical standards for human testing, and all subjects must be tested voluntarily and sign an informed consent form before the test. Before the subject signs the informed consent form, the tester needs to inform the subject of the purpose of the test, possible benefits, potential risks and problems, and related rights and obligations.

[0250] 2.1.1 Target Number of People Target number of subjects: 30.

[0251] 2.1.2 Selection criteria Age range: 18 to 60 years old, no gender restrictions.

[0252] Health status: Male or female in good health.

[0253] Hair problems: At least one of the symptoms of dandruff and oily hair, or excessive hair loss or thinning hair (the hair loss count is greater than 10 when combing the hair 60 times).

[0254] Hair treatment: No hair dyeing, perming or other hair treatments have been performed in the past month.

[0255] Informed consent: understand the testing process, voluntarily participate in the test, and sign the informed consent form.

[0256] Follow-up visit requirements: Able to conduct follow-up visits as required.

[0257] 2.1.3 Exclusion criteria Pregnant or breastfeeding women, and those who are planning to get pregnant in the near future.

[0258] People with severe androgenic alopecia, alopecia areata, inflammatory scar alopecia or other pathological hair loss.

[0259] People who have used anti-hair loss cosmetics or other products with similar effects in the past 3 months.

[0260] People who have undergone hair transplantation treatment.

[0261] People with naturally curly hair.

[0262] People with highly sensitive constitutions.

[0263] People who are currently participating in or have participated in other clinical trials or hair care product tests within the past 2 months.

[0264] People with severe systemic diseases or other skin diseases (such as lupus erythematosus, psoriasis, etc.).

[0265] People who frequently change their shampoo products on a daily basis.

[0266] People whose scalp has been treated with medication.

[0267] Non-voluntary participants or people who cannot complete the required content according to the test requirements.

[0268] 2.1.4. Exit (dropout) criteria The subject is lost to follow-up or actively requests to withdraw from the trial; The subjects had poor compliance and could not attend the follow-up visit as required; The subjects used other products similar to the test samples during the study; The subject develops new diseases during the entire study period that directly affect the evaluation of clinical status, including skin diseases; The subject developed severe illness during the entire study period; The subject became pregnant during the study; Subjects experienced serious adverse reactions throughout the study period.

[0269] 2.2 Test site 2.2.1. For the test indicators of dandruff quantity and hair loss quantity, the test site is the whole head, and the left and right partitions are used for self-control.

[0270] 2.2.2 For the scalp surface oil content and hair density test indicators, the test site is to select three 1.5 cm × 1.5 cm test areas on the left and right sides of the sparse hair area on the top of the head, and the distance between these areas is at least 5 cm. Starting from the position closest to the forehead, the test areas are numbered in a clockwise direction. According to the random table, the test is carried out at the respective numbered positions.

[0271] 2.3 Test environment The subjects were required to acclimate for 30 minutes at a temperature maintained at 20-22°C and relative humidity maintained at 40%-60%, followed by evaluation and measurement.

[0272] 2.4 Number of people A total of 31 subjects participated, including 8 males and 23 females, with an average age of 45.0±11.0 years. Due to the problem of follow-up time, 1 subject withdrew from the study, and finally 30 subjects completed the 8-week test.

[0273] 3. Test process 3.1 Researcher Education Provide research plans, explain the research content to subjects, obtain informed consent, arrange follow-up and care during the study, enhance subject compliance, and introduce possible adverse reactions of research drugs and countermeasures.

[0274] 3.2 Pre-screening All volunteers will be informed of the inclusion and exclusion criteria by phone. After eligible volunteers arrive at the test center, they will be further screened by researchers. Volunteers can only be formally enrolled in the test after meeting the inclusion and exclusion criteria and signing the informed consent form. On the day of the test, volunteers are not allowed to engage in any other activities that may affect the screening evaluation results.

[0275] 3.3 Enrollment Subjects were included in the study if they met the inclusion criteria. During the two-day wash-out period, no skin care or washing products were used on the scalp. On the day of the test, the subjects were required to sit quietly in a laboratory-controlled environment (temperature 22±1℃, relative humidity 50%±10%) for 30 minutes before data collection.

[0276] 3.4 Data Collection The subjects underwent scalp and hair tests before using the samples and after using the products for 2, 4, and 8 weeks. The tests were conducted approximately 48 hours after shampooing, and the subjects needed to adapt to the test environment for 30 minutes before each test. The hair was combed 30 times from the left front to the left back and from the right front to the right back, and the shed hair and dandruff were collected and counted. A 1.5cm×1.5cm area was selected on each side as the test area, and the hair in the area was cut to within 1mm and as close to the scalp as possible. The two test areas were imaged using a scalp analyzer. After the test, the hair density and oil status were analyzed.

[0277] 4. Adverse reactions If any adverse reaction occurs during the use of the test product, the volunteer should stop using the product immediately and go to the research center for a doctor's examination to decide whether to continue using the product. At the same time, all adverse reactions and related events during the test must be recorded in detail and reflected in the final report.

[0278] 5. Data Analysis SPSS 22.0 software was used for data analysis. The measurement data were expressed as mean ± standard deviation and tested for normal distribution. If the data conformed to normal distribution, paired t test was used for self-comparison before and after; if the data did not conform to normal distribution, two related sample rank sum test was used. Two related sample rank sum test was used for before and after comparison of graded data; independent sample t test or rank sum test was used for comparison between the test product and the control group. All statistical analyses were two-tailed tests, and the significance level was set at α = 0.05. The significant differences in scalp and hair parameters before and after using the samples were analyzed by paired T test, with a significance level of α = 0.05; the significant differences in the subjects' satisfaction with the scalp status before and after using the product were analyzed by the chi-square test of Fisher's exact probability method, with a significance level of α = 0.05.

[0279] 6. Experimental results A total of 31 subjects participated, including 8 males and 23 females, with an average age of 45.0±11.0 years. Due to the time of the follow-up visit, 1 subject withdrew from the study, and finally 30 subjects completed the 8-week test. The subjects were tested by the staff three times a week on half of their heads. The amount of hair loss, hair density, dandruff, and scalp surface oil content were measured before using the product, 2 weeks, 4 weeks, and 8 weeks after using the product. The test results are as follows: 6.1. Evaluation of the effect of Platycladus orientalis shampoo on the amount of hair loss in subjects The subjects were not allowed to wash their hair during the study period and were not allowed to comb their hair on the morning of the visit. After arriving at the laboratory, the same technician used a comb of uniform specifications to comb the hair 30 times on each side in the order of left front to left back and right front to right back. Afterwards, the hair that fell off on both sides was collected and counted.

[0280] The test results of the amount of hair loss at various time points before and after using the product are shown in Table 24 (unit: roots / half head).

[0281] Table 24

[0282] Note: Conduct a significance analysis before using the product. p<0.05, p<0.01.

[0283] The trend of hair loss at each time point before and after using the product is shown in Figure 1 . Conduct significance analysis on test samples and controls before using the product. p<0.05, p<0.01.

[0284] The trend of the difference between the amount of hair loss at each time point after using the product and before using the product is shown in Figure 2 . Conduct a significance analysis before using the product. p<0.01.

[0285] Compared with before using the product, there was no significant difference in the amount of hair loss at each time point in the control area (p≥0.05); there was no significant difference in the area using the test sample in the second week (p≥0.05), a significant decrease in the fourth week (p<0.05), and a very significant decrease in the eighth week (p<0.01).

[0286] Compared with the control hair loss amount measurement value, there was no significant difference in the amount of hair loss in the test sample area before using the product (p≥0.05), but it was significantly reduced in the 2nd week (p<0.05), and extremely significantly reduced in the 4th and 8th weeks (p<0.01).

[0287] Compared with the change in the number of hair loss in the control group, the change in the number of hair loss using the test sample was significantly reduced at the 2nd, 4th and 8th weeks (p<0.01).

[0288] 6.2 Evaluation of the effect of Platycladus orientalis shampoo on hair density of subjects The hair in the test area needs to be cut to less than 1 mm. Use the scalp analyzer of South Korea's KC company to image the scalp and hair in the test area and count the number of hairs in the photo. At the same time, measure the area of ​​the scalp area according to the scale in the photo. And calculate the hair density according to the following formula. The higher the hair density D value, the denser the hair per unit area: D=N÷S.

[0289] Note: N is the number of hair roots, S is the scalp area (cm 2 ), D represents the hair density (roots / cm 2 ).

[0290] The test results of hair density at various time points before and after using the product are shown in Table 25 (Unit: roots / cm 2 ).

[0291] Table 25

[0292] Note: Conduct a significance analysis before using the product. p<0.05, p<0.01.

[0293] The trend of hair density changes at various time points before and after using the product is shown in Figure 3 The trend of the difference in hair density between each time point after using the product and before using the product is shown in Figure 4 . Conduct a significance analysis before using the product. p<0.05, p<0.01.

[0294] Compared with before using the product, there was no significant difference in hair density at each time point in the control area (p≥0.05). The hair density in the area using the test sample increased significantly in the second week (p<0.05), and increased extremely significantly in the fourth and eighth weeks (p<0.01).

[0295] Compared with the hair density measurements of the control, there was no significant difference in hair density in the test sample area before using the product (p≥0.05), but it increased significantly at the 2nd and 4th weeks (p<0.05), and extremely significantly at the 8th week (p<0.01).

[0296] Compared with the hair density change value of the control, the hair density change value of the test sample increased significantly in the 2nd and 4th weeks (p<0.05), and the hair density change value increased extremely significantly in the 8th week (p<0.01).

[0297] 6.3 Evaluation of the effect of scalp essence containing Platycladus orientalis leaf extract on the amount of dandruff The subjects were prohibited from washing their hair by themselves during the study period, and were not allowed to comb their hair on the morning of the visit. After arriving at the laboratory, the same technician used a comb of uniform specifications to comb the hair 30 times on each side in the order from left front to left back and right front to right back to collect dandruff on a black plastic shell, and then took pictures of the collected dandruff with a SLR camera, and used ImageJ to count the number of dandruff in the photos. The number of dandruff is measured in "grains", and the larger the value, the more serious the dandruff problem.

[0298] The test results of the number of dandruffs before and after the subjects used the product are shown in Table 26 (unit: grains).

[0299] Table 26

[0300] Note: Conduct a significance analysis before using the product. p<0.05, p<0.01.

[0301] The test results of the number of dandruff before and after using the product are shown in Figure 5 :The trend of the difference between the number of dandruff at each time point after using the product and before using the product is shown in Figure 6 . Conduct a significance analysis before using the product. p<0.01.

[0302] Compared with before using the product, there was no significant difference in the amount of dandruff in the control area at each time point (p≥0.05), and the amount of dandruff in the area using the test sample was significantly reduced in the 2nd week, 4th week, and 8th week (p<0.01).

[0303] Compared with the dandruff count of the control, the dandruff count in the area using the test sample was significantly reduced before using the product, at the 4th week, the 8th week, and the 12th week (p<0.01).

[0304] Compared with the change value of the dandruff quantity of the control, the change value of the dandruff quantity in the 2nd, 4th and 8th week of using the test sample was extremely significantly reduced (p<0.01).

[0305] 6.4 Evaluation of the effect of scalp essence containing Platycladus orientalis leaf extract on the oil content on the scalp surface The scalp oil content test is completed by the SM815 probe of the MPA 580 skin tester of the German CK company. The probe contains a special matte tape with a width of 8 mm and a thickness of 0.1 mm. When this tape comes into contact with the oil on the skin, it will gradually become translucent and the light transmittance will increase. The amount of light transmittance directly reflects the oil content: the more oil, the greater the light transmittance. The test result range is 0-350 μg / cm 2 The higher the value, the more oil there is on the scalp.

[0306] The test results of the oil content on the scalp surface of the subjects before and after using the product are shown in Table 27 (unit: μg / cm 2 ).

[0307] Table 27

[0308] Note: Conduct a significance analysis before using the product. p<0.05, p<0.01.

[0309] The test results of the oil content on the scalp surface of the subjects before and after using the product are shown in Figure 7 The trend of the difference in scalp surface oil content at each time point after using the product and before using the product is shown in Figure 8 . Conduct a significance analysis before using the product. p<0.01.

[0310] Compared with before using the product, there was no significant difference in the oil content on the scalp surface at each time point in the control area (p≥0.05). The oil content in the test sample area was significantly reduced in the 2nd week (p<0.05), and was extremely significantly reduced in the 4th and 8th weeks (p<0.01).

[0311] Compared with the control scalp surface oil content, the scalp surface oil content in the area using the test sample was significantly reduced in the 2nd, 4th and 8th weeks of using the product (p<0.01).

[0312] Compared with the change value of the control scalp surface oil content, the change value of the scalp surface oil content in the 2nd week, 4th week, and 8th week of using the test sample was extremely significantly reduced (p<0.01).

[0313] In summary, shampoo containing Thuja orientalis leaf samples can significantly reduce the amount of dandruff, while shampoo without Thuja orientalis leaf samples has no significant effect on the amount of dandruff, indicating that shampoo containing Thuja orientalis leaf samples has an anti-dandruff effect; shampoo containing Thuja orientalis leaf samples can significantly reduce the oil content on the scalp surface, while shampoo without Thuja orientalis leaf samples has no significant effect on the oil content on the scalp surface, indicating that shampoo containing Thuja orientalis leaf samples has an oil control effect. Scalp essence containing Thuja orientalis leaf samples can significantly reduce the amount of hair loss, while scalp essence without Thuja orientalis leaf samples has no significant effect on the amount of hair loss, indicating that scalp essence containing Thuja orientalis leaf samples has an anti-hair loss effect; scalp essence containing Thuja orientalis leaf samples can significantly increase hair density, while scalp essence without Thuja orientalis leaf samples has no significant effect on hair density, indicating that scalp essence containing Thuja orientalis leaf samples has a hair growth and hair strengthening effect.

[0314] Example 7 A mud mask containing Platycladus orientalis leaf extract. The formula and preparation process are shown in Table 28.

[0315] Table 28

[0316] Among them, the Platycladus orientalis leaf pulp is the Platycladus orientalis leaf pulp obtained in step S602 in Example 1.

[0317] Comparative Example 9 A mud mask without Platycladus orientalis leaf extract, the formula and preparation process of which are shown in Table 29.

[0318] Table 29

[0319] The content of microcrystalline cellulose in the microcrystalline cellulose slurry is 25%, and the particle size is 10-30 μm.

[0320] Effect evaluation 7 Consumer reviews of Platycladus orientalis mud mask.

[0321] 1. Test methods 1.1. Usage method: The staff used the Platycladus orientalis leaf mud mask sample prepared in Example 7 for the subjects for 3 times on Day 0, Day 3, and Day 6, respectively, and used it on the same half face (test sample group). The control mud mask prepared in Comparative Example 9 was used on the other side as a control (control group). After the subjects cleaned their faces, they took an appropriate amount of the mud mask and evenly applied it on the face, left it for 15 minutes, and then washed it off.

[0322] 1.2. Instruments and software: Multifunctional skin tester MPA580 (CK, Germany), ImageJ 1.52a (NIH, USA).

[0323] 1.2.1. Skin surface oil content test The skin oil content test is completed by the SM815 probe of the MPA 580 skin tester of the German CK company. The probe contains a special matte tape with a width of 8 mm and a thickness of 0.1 mm. When this tape comes into contact with the oil on the skin, it will gradually become translucent and the light transmittance will increase. The amount of light transmittance directly reflects the oil content: the more oil, the greater the light transmittance. The test result range is 0-350 μg / cm 2 The higher the value, the more oil there is in the epidermis.

[0324] 1.2.2. Acne Quantity Detection During the study period, the subjects were prohibited from using acne removal products on their own, and were not allowed to use any acne removal tools such as comedone needles. During the follow-up period, the same experimenter performed subjective evaluation by counting the comedones on the left and right cheeks of the subjects. The lower the manual comedone count value, the fewer the number of comedones.

[0325] 1.2.3. Skin stratum corneum moisture content detection The capacitance method is used to measure the moisture content of the skin surface. The measurement principle of the capacitance method is based on the fact that the dielectric constant of water in the skin surface is much higher than that of other substances. Therefore, the change in the dielectric constant of the skin surface is mainly affected by the moisture content of the skin surface. By measuring the dielectric constant of the skin surface, the moisture content of the skin surface can be analyzed. The measured value is a relative value in CU (Corneometer Units), which is specifically used to indicate the moisture content of the skin. In the measurement results, the higher the value, the richer the moisture content of the skin surface.

[0326] 2. Test conditions 2.1. Requirements for Subjects According to the Declaration of Helsinki, the selection of subjects follows the medical and ethical standards for human testing, and all subjects must be tested voluntarily and sign an informed consent form before the test. Before the subject signs the informed consent form, the tester needs to inform the subject of the purpose of the test, possible benefits, potential risks and problems, and related rights and obligations.

[0327] Target number of subjects: 15.

[0328] 2.2. Selection Criteria Age between 18 and 45, healthy women or men.

[0329] Those with a certain number of acne on their facial skin cannot have nodules or cysts that are difficult to improve with clinical preparations. Trained experimenters graded the severity of acne on the subjects according to the facial acne grading standard (refer to Pillsbury and the International Modified Acne Grading System), and subjects with mild grade I and moderate grade II were selected.

[0330] Informed consent: understand the testing process, voluntarily participate in the test, and sign the informed consent form.

[0331] Follow-up visit requirements: Able to conduct follow-up visits as required.

[0332] 2.3 Exclusion criteria Pregnant or breastfeeding women, and those who are planning to get pregnant in the near future.

[0333] People with allergic or highly sensitive constitution.

[0334] Have a history of allergy to cosmetics or other external preparations.

[0335] People with allergic skin diseases have a history of skin diseases or diseases, such as eczema, psoriasis, atopic dermatitis, severe acne, tinea, pityriasis, severe seborrheic dermatitis, bacterial or viral infections, etc.

[0336] People who are receiving dermatological treatment.

[0337] The test area has undergone cosmetic surgery, laser or other cosmetic procedures that may affect the test results in the past six months.

[0338] The test area has skin manifestations that may affect the test results, such as birthmarks, scratches, white spots, pigmentation, moles, atrophy, scars or other blemishes.

[0339] Use of antihistamines in the past week or hormonal drugs and immunosuppressants in the past month.

[0340] Those who have used any anti-inflammatory drugs on the test area in the past two months.

[0341] Patients with severe systemic diseases, immunodeficiency or autoimmune diseases.

[0342] Patients with insulin-dependent diabetes mellitus.

[0343] People who are being treated for asthma or other respiratory problems.

[0344] Bilateral mastectomy and bilateral axillary lymph node removal.

[0345] Those who have received anti-cancer chemotherapy in the past six months.

[0346] Other patients deemed unsuitable for trial participation by clinical assessment.

[0347] Those who are unwilling to participate or are unable to complete the required content according to the test requirements.

[0348] 2.4. Exit (dropout) criteria The subject is lost to follow-up or actively requests to withdraw from the trial; The subjects had poor compliance and could not attend the follow-up visit as required; The subjects used other products similar to the test samples during the study; The subject develops new diseases during the entire study period that directly affect the evaluation of clinical status, including skin diseases; The subject developed severe illness during the entire study period; The subject became pregnant during the study; Subjects experienced serious adverse reactions throughout the study period.

[0349] 2.5 Test Location Face, compare the left and right sides of the face to yourself.

[0350] 2.6 Test Environment The subjects were required to acclimate for 30 minutes at a temperature maintained at 20-22°C and relative humidity maintained at 40%-60%, followed by evaluation and measurement.

[0351] 2.7 Number of people A total of 17 subjects participated, including 10 males and 7 females, with an average age of 23.6±4.0 years. Due to the problem of follow-up time, 2 subjects withdrew from the study, and finally 15 subjects completed the 1-week test.

[0352] 3. Test process 3.1 Researcher Education Provide research plans, explain the research content to subjects, obtain informed consent, arrange follow-up and care during the study, enhance subject compliance, and introduce possible adverse reactions of research drugs and countermeasures.

[0353] 3.2 Pre-screening All volunteers were informed of the inclusion and exclusion criteria by phone. After eligible volunteers arrived at the test center, they were further screened by researchers. Trained experimenters graded the severity of the subjects' acne according to the facial acne grading standard. Subjects with mild grade I and moderate grade II were selected for testing and recorded. Volunteers could only be formally enrolled in the test after meeting the inclusion and exclusion criteria and signing the informed consent form.

[0354] 3.3 Enrollment Subjects who met the inclusion criteria were included in the study. Subjects were not allowed to use any products (cosmetics or topical drugs) on their facial area on the day of the test. On the day of the test, the experimenter guided the subjects to clean their facial skin with facial cleanser. Subjects were required to sit quietly in a laboratory-controlled environment (temperature 22±1℃, relative humidity 50%±10%) for 30 minutes before data collection.

[0355] 3.4 Data Collection Before using the product on Day 0, the experimenter manually counted the comedones on the subjects and used a skin tester to test the epidermal oil content and the moisture content of the stratum corneum on both sides of the face; 30 minutes after using the product on Day 6, the experimenter manually counted the comedones on the subjects and used a skin tester to test the epidermal oil content and the moisture content of the stratum corneum on both sides of the face.

[0356] 4. Adverse reactions If any adverse reaction occurs during the use of the test product, the volunteer should stop using the product immediately and go to the research center for a doctor's examination to decide whether to continue using the product. At the same time, all adverse reactions and related events during the test must be recorded in detail and reflected in the final report.

[0357] 5. Data Analysis SPSS 22.0 software was used for data analysis. The measurement data were expressed as mean ± standard deviation and tested for normal distribution. If the data conformed to normal distribution, paired t test was used for self-comparison before and after; if the data did not conform to normal distribution, two related sample rank sum test was used. Two related sample rank sum test was used for before and after comparison of graded data; independent sample t test or rank sum test was used for comparison between test product and control group. All statistical analyses were two-tailed tests, with significance level set at α = 0.05. Paired T test was used to analyze the significant difference of the face before and after the sample was used, with significance level α = 0.05.

[0358] 6. Experimental results 6.1 Effect of Platycladus orientalis mud mask on skin surface oil content The test of skin surface oil content is completed by the SM815 probe of the MPA 580 skin tester of the German CK company. The probe contains a special matte tape with a width of 8 mm and a thickness of 0.1 mm. When this tape comes into contact with the oil on the skin, it will gradually become translucent and the light transmittance will increase. The amount of light transmittance directly reflects the oil content: the more oil, the greater the light transmittance. The test result range is 0-350 μg / cm 2 The higher the value, the more oil there is on the skin surface.

[0359] The test results of the skin surface oil content of the subjects before and after using the product are shown in Table 30 (unit: μg / cm 2 ).

[0360] Table 30

[0361] Note: Conduct a significance analysis before using the product. p<0.05.

[0362] The test results of the skin surface oil content of the subjects before and after using the product are shown in Fig. 9 ; The difference in skin surface oil content between before and after using the product is shown in Fig.10 . A significant difference analysis was performed with the control group. p<0.01.

[0363] Compared with before using the product, there was no significant difference in the skin surface oil content in the control area (p≥0.05), and it was significantly reduced in the test sample area after 1 week of use (p<0.05).

[0364] Compared with the skin surface oil content of the control, the skin surface oil content of the test sample area was significantly reduced after using the product for 1 week (p<0.01).

[0365] Compared with the change value of skin surface oil content in the control group, the change value of skin surface oil content after using the test sample for 1 week was extremely significantly reduced (p<0.01).

[0366] 6.2 Effect of Platycladus orientalis mud mask on the number of acne During the study period, the subjects were prohibited from using acne removal products on their own, and were not allowed to use any acne removal tools such as comedone needles. During the follow-up period, the same experimenter performed subjective evaluation by counting the comedones on the left and right cheeks of the subjects. The lower the manual comedone count value, the fewer the number of comedones.

[0367] The results of the number of acne on the subjects before and after using the product are shown in Table 31 (unit: pieces).

[0368] Table 31

[0369] Note: Conduct a significance analysis before using the product. p<0.01.

[0370] The test results of the number of acne before and after the subjects used the product are shown in Fig.11 ; The difference in the number of acne after using the product and before using the product is shown in Fig.12 . A significant difference analysis was performed with the control group. p<0.01.

[0371] There was no significant difference in the number of acne in the control area compared with before using the product (p≥0.05), but it was significantly reduced in the test sample area after 1 week of use (p<0.01).

[0372] Compared with the number of acne in the control, the number of acne in the area using the test sample was significantly reduced after 1 week of using the product (p<0.01).

[0373] Compared with the change in the number of acne in the control, the change in the number of acne after using the test sample for 1 week was extremely significantly reduced (p<0.01).

[0374] 6.3 Effect of Platycladus orientalis mud mask on moisture content of skin stratum corneum The test was conducted using the multifunctional skin tester MPA 580 (equipped with a Corneometer CM 825 probe) from the German CK company. The capacitance method was used to measure the moisture content of the skin surface. The measurement principle of the capacitance method is based on the fact that the dielectric constant of water in the skin surface is much higher than that of other substances. Therefore, the change in the dielectric constant of the skin surface is mainly affected by the moisture content of the skin surface. By measuring the dielectric constant of the skin surface, the moisture content of the skin surface can be analyzed. The measured value is a relative value in CU (Corneometer Units), which is a unit specifically used to indicate the moisture content of the skin. In the measurement results, the higher the value, the richer the moisture content of the skin surface.

[0375] The results of the water content of the skin stratum corneum of the subjects before and after using the product are shown in Table 32 (unit: CU).

[0376] Table 32

[0377] Note: Conduct a significance analysis before using the product. p<0.05.

[0378] The test results of the water content of the skin stratum corneum of the subjects before and after using the product are shown in Fig.13 ; The difference in water content of the skin stratum corneum before and after using the product is shown in Fig.14 . A significant difference analysis was performed with the control group. p<0.01.

[0379] Compared with before using the product, there was no significant difference in the water content of the skin stratum corneum in the control area (p≥0.05), but it increased significantly after 1 week of use in the test sample area (p<0.05).

[0380] Compared with the stratum corneum water content of the control, the stratum corneum water content of the skin in the area using the test sample increased significantly after using the product for 1 week (p<0.01).

[0381] Compared with the change in water content of the skin stratum corneum of the control, the change in water content of the skin stratum corneum after using the test sample for 1 week increased significantly (p<0.01).

[0382] In summary, the mud mask containing Thuja orientalis leaf pulp can significantly reduce the number of acne, while the mud mask without Thuja orientalis leaf pulp has no significant effect on the number of acne, indicating that the mud mask containing Thuja orientalis leaf pulp has the effect of removing acne; the mud mask containing Thuja orientalis leaf pulp can significantly reduce the oil content on the skin surface, while the mud mask without Thuja orientalis leaf pulp has no significant effect on the oil content on the skin surface, indicating that the mud mask containing Thuja orientalis leaf pulp has the effect of oil control; the mud mask containing Thuja orientalis leaf pulp can significantly increase the water content of the skin stratum corneum, while the mud mask without Thuja orientalis leaf pulp has no significant effect on the water content of the skin stratum corneum, indicating that the mud mask containing Thuja orientalis leaf pulp has the effect of hydrating and moisturizing.

[0383] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing a Platycladus orientalis leaf extract, characterized in that: The following steps are involved: S1: Mix the Platycladus orientalis leaves with 4-16 times the weight of water, perform cyclic percolation extraction, repeat 2-4 times, combine the filtrates, and obtain the Platycladus orientalis leaf cyclic percolation liquid and medicinal residues; S2: Concentrating the circulating leachate of Platycladus orientalis leaves obtained in step S1 through a filter membrane to a concentration of 5-7 times the weight of the Platycladus orientalis leaves in step S1, and filtering to obtain a Platycladus orientalis leaf concentrated filtrate; S3: performing vacuum distillation to concentrate the concentrated filtrate of Platycladus orientalis leaves obtained in step S2, and simultaneously collecting the concentrated solution and condensate obtained by the vacuum distillation, wherein the condensate is Platycladus orientalis hydrosol; S4: cooling the concentrated solution obtained in step S3, mixing it with an ethanol aqueous solution to obtain a mixed solution, cooling it to -6 to 6°C, letting it stand, filtering it, filtering it with a molecular interception membrane, concentrating it, and drying it to obtain a Platycladus orientalis leaf extract; S5: grinding the medicinal residue obtained in step S1 to obtain Platycladus orientalis slurry; Wherein, in step S4, the volume fraction of ethanol in the mixed solution is 20%-30%; the standing time is ≥2h; and the molecular retention membrane filtration is performed using a molecular retention membrane filtration of 1000-5000 Daltons.

2. The preparation method according to claim 1, characterized in that: In step S1, the mass content of quercetin in the Platycladus orientalis leaves is ≥0.25%.

3. The preparation method according to claim 1, characterized in that: In step S1, each cyclic diafiltration extraction comprises the following steps: cyclic diafiltration at 80-99°C for 30-90 minutes, filtering, and cooling the filtrate to ≤60°C.

4. The preparation method according to claim 3, characterized in that: In step S1, the cyclic diafiltration extraction is repeated 3 times. The first extraction comprises the following steps: mixing Platycladus orientalis leaves with 10 times the weight of water, performing cyclic percolation extraction at 90-99° C. for 55-65 minutes, filtering, rapidly cooling the filtrate to below 60° C., and collecting the filter residue; The second extraction comprises the following steps: adding 6 times of water to the residue obtained in the first extraction step, performing cyclic percolation extraction at 90-99°C for 55-65 minutes, filtering, rapidly cooling the filtrate to below 60°C, and collecting the residue; The third extraction comprises the following steps: adding 4 times of water to the filter residue obtained in the first extraction step, cyclic percolation extraction at 90-99° C. for 55-65 minutes, filtering, rapidly cooling the filtrate to below 60° C., and collecting the filter residue.

5. The preparation method according to claim 1, characterized in that: In step S2, the membrane concentration is performed by using a 100-300 Dalton membrane, and the filtration is performed until the solution is clarified.

6. The preparation method according to claim 1, characterized in that: In step S3, the temperature of the reduced pressure distillation concentration is ≤70°C, and the weight of the concentrated liquid is 0.8-1.4 times the weight of the Platycladus orientalis leaves in step S1.

7. The preparation method according to claim 1, characterized in that: In step S4, the volume fraction of ethanol in the ethanol aqueous solution is 90%-99%.

8. The preparation method according to claim 1, characterized in that: In step S5, the refining is specifically performed by using a colloid mill to refine the pulp until the average solid particle size is 10-30 μm.

9. The preparation method according to claim 1, characterized in that: The step S601 is also included: S601: Dry the Platycladus orientalis slurry obtained in step S5 to obtain Platycladus orientalis leaf powder.

10. The preparation method according to claim 1, characterized in that: The step S602 is also included: S602: Concentrate the Platycladus orientalis slurry obtained in step S5 to a solid weight content of 20%-35% to obtain Platycladus orientalis leaf pulp.

11. The Platycladus orientalis leaf extract obtained by the preparation method according to any one of claims 1 to 10.

12. The Platycladus orientalis hydrosol obtained by the preparation method described in any one of claims 1 to 10.

13. The Platycladus orientalis slurry obtained by the preparation method described in any one of claims 1 to 10.

14. The Platycladus orientalis leaf powder obtained by the preparation method according to claim 9.

15. The Platycladus orientalis leaf pulp obtained by the preparation method described in claim 10.

16. Use of the preparation method according to any one of claims 1 to 10 in the production of Platycladus orientalis leaf extract.

17. Use of the Platycladus orientalis leaf extract according to claim 11, the Platycladus orientalis hydrosol according to claim 12, the Platycladus orientalis slurry according to claim 13, the Platycladus orientalis leaf powder according to claim 14, or the Platycladus orientalis leaf slurry according to claim 15 in cosmetics.

18. The use according to claim 17, characterized in that The cosmetic is selected from at least one of the following: (1) Cosmetics that inhibit sebaceous gland oil secretion; (2) Cosmetics that promote the expression of FLG and Caspase14 in keratinocytes; (3) Cosmetics that inhibit oil secretion and promote oil-water balance; (4) Cosmetics that treat acne, prevent acne, replenish moisture, and inhibit oil secretion.

19. The use according to claim 17, characterized in that: The cosmetics are cosmetics that have the functions of promoting the expression of at least one gene among ELOVL4, ELOVL7 and CerS4, inhibiting oil secretion and promoting oil-water balance.

20. A cosmetic, characterized in that: The ingredients include the Platycladus orientalis leaf extract according to claim 11, the Platycladus orientalis pure water according to claim 12, the Platycladus orientalis slurry according to claim 13, the Platycladus orientalis leaf powder according to claim 14, or the Platycladus orientalis leaf slurry according to claim 15.

21. A shampoo, characterized in that: Includes the following ingredients: Water, sodium laureth sulfate, laureth-2, sodium sulfate, Platycladus orientalis leaf extract, glycerin, polyquaternium-10, sodium cocoamidopropionate, sodium chloride, cocamidopropyl betaine, citric acid, cocamide MEA, phenoxyethanol and pH adjuster; Wherein, the Platycladus orientalis leaf extract is the Platycladus orientalis leaf extract according to claim 11.

22. A scalp essence, characterized in that: Includes the following ingredients: Water, polyacrylate crosspolymer-6, Platycladus orientalis leaf extract, 1,2-hexanediol, butylene glycol, sodium metabisulfite, and arginine; Wherein, the Platycladus orientalis leaf extract is the Platycladus orientalis leaf extract according to claim 11.

23. A Platycladus orientalis leaf mud film, characterized in that: Includes the following ingredients: Water, polyacrylate crosspolymer-6, 1,2-hexanediol, p-hydroxyacetophenone, arginine, glycerin and Thuja orientalis leaf pulp; Wherein, the Platycladus orientalis leaf pulp is the Platycladus orientalis leaf pulp described in claim 15.

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