Composition with skin basement membrane repairing effect

Abstract

The invention provides a composition containing a rhodiola rosea supercritical fluid extract, a rhodiola rosea alcohol extract and a rhodiola rosea polysaccharide extract. The invention also relates to a rhodiola rosea supercritical fluid extract. The present invention further relates to a composition comprising any two selected from the group consisting of a supercritical fluid extract of Rhodiola rosea, an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea. The invention further relates to application of the composition or the rhodiola rosea supercritical fluid extract, the rhodiola rosea alcohol extract or the rhodiola rosea polysaccharide extract in preparation of cosmetics or medicines. The invention further relates to the cosmetics or the medicines containing the composition or the rhodiola rosea supercritical fluid extract.

Description

Technical Field

[0001] The present invention relates to a composition comprising a supercritical fluid extract of Rhodiola rosea, an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea. The present invention also relates to a supercritical fluid extract of Rhodiola rosea, a composition comprising any two components of a supercritical fluid extract of Rhodiola rosea, an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea. Use of the composition or the supercritical fluid extract of Rhodiola rosea, the alcohol extract of Rhodiola rosea or the polysaccharide extract of Rhodiola rosea in the preparation of cosmetics or medicines, and cosmetics or medicines containing the composition or the supercritical fluid extract of Rhodiola rosea. Background Art

[0002] The skin is a protective barrier that protects the body from environmental aggressions. The outermost layer of the skin is the epidermis (epithelium), which is separated from the dermis (mesenchyme) by a thin basement membrane (BM) and anchoring proteins. The basement membrane is similar to a sheet-like matrix and contains highly organized extracellular matrix (ECM) proteins, which together with anchoring proteins form the dermal-epidermal junction (DEJ). The skin basement membrane (DEJ-BM) distinguishes the dermis and epidermis of the skin. It tightly binds the epidermis to the dermis and provides shear resistance. It acts as a permeability barrier and provides an interface for nutrient storage and the conduction of various signal molecules. These signal transduction activities affect tissue growth, guide cell migration, and support cell survival and differentiation.

[0003] Based on their appearance under transmission electron microscopy (TEM), the DEJ-BM is subdivided into three morphological layers: The stratum lucidum, located above the basement membrane, is an electron-lucent zone just below the plasma membrane of basal keratinocytes. It contains anchoring filaments, i.e., anchoring proteins (e.g., integrin α6β4, laminin-322, collagen type VII, and collagen type XVII) that play a key role in strengthening the dermal-epidermal junction. Adjacent to the stratum compactum, keratinocytes insert hemidesmosomes into an intermediate filament network characterized by the expression of integrin α6β4 and collagen type XVII. Although epidermal keratinocytes and dermal fibroblasts synthesize DEJ-BM proteins, collagen type XVII is primarily synthesized by keratinocytes, while nestin is primarily synthesized by fibroblasts. Collagen type XVII plays an important role in the assembly and function of cell-matrix adhesion structures, as well as in transmembrane signaling and keratinocyte differentiation. Laminin, as the most abundant glycoprotein in the basement membrane ECM, also plays an important role in supporting tissue architecture and stability. Laminins participate in skin epithelialization and wound healing by regulating core cell behaviors such as adhesion, proliferation, migration, apoptosis, and differentiation. The lamina compacta, named for its electron-dense appearance in TEM, is formed by laminin-511 / type IV collagen polymers. On its underside, the DEJ-BM is connected to the dermis via a ring-like structure of type VII collagen, linking laminin-332 in the lamina compacta to type I and type III collagen fibers in the dermis. Nestin is assembled into the basement membrane ECM together with perlecan, acting as a bridging protein between type IV collagen and laminin to form an intact skin basement membrane.

[0004] The complex network of interconnected DEJ-BM proteins and finger-like projections help provide structural integrity and mechanical stability to the skin. However, with intrinsic skin aging, the DEJ-BM becomes significantly thinner and takes on a flat appearance due to the loss of epidermal ridges, which leads to impaired adhesion of the epidermis to the dermis and reduced interfacial area for the movement of nutrients and signaling molecules. As a result, aged skin has poor resistance to shear forces and is more susceptible to injury and wrinkling. Inherited or acquired defects in DEJ-BM-related components can lead to severe or fatal blistering skin diseases. Therefore, based on the important role of DEJ-BM in skin homeostasis, orderly regulation of basement membrane protein expression is of great value in skin wound healing and delaying skin aging in regenerative medicine.

[0005] At present, there are many studies on basement membrane repair to achieve skin anti-wrinkle and firming effects and repair epidermal barrier function. For example, authorization number CN116763677B discloses a basement membrane repair composition (containing acetyl tetrapeptide-9, acetyl tetrapeptide-11 and polyols, etc.), which has good transdermal effect, mild and non-irritating, and has basement membrane repair effect and anti-wrinkle effect. Application number CN117618309A discloses an anti-aging composition, which is designed from the perspective of epidermal barrier repair, DEJ repair, dermal repair, and mitochondrial ROS removal, taking into account the epidermis, dermis, and muscle layer for multi-dimensional skin anti-aging. Application number CN108283617A discloses a composition composed of keratin, hydrolyzed royal jelly, and glycerophosphocholine, which can prolong cell life, delay skin aging, promote basement membrane zone cell growth and repair, thereby maintaining the skin barrier from the inside out, but lacks experimental data support related to in vitro cells, and only partially subjective partial human efficacy test survey data.

[0006] Rhodiola rosea plants are an important part of traditional Chinese medicine and Tibetan medicine. They are natural precious medicinal materials with broad application prospects. The plants of this genus contain key compounds such as ginsenosides, polysaccharides and flavonoids, and have anti-inflammatory, antioxidant, anti-aging, hypoglycemic, immune-enhancing and anti-hypoxia properties. Rhodiola rosea extracts are mainly used as antioxidants, skin conditioners, and whitening and anti-freckle agents in cosmetics and skin care products. However, there has been no discovery or report of Rhodiola rosea extracts or Rhodiola rosea-related compositions in the field of skin basement membrane repair.

[0007] Therefore, there is an urgent need for a novel compound / composition / plant extract having excellent skin basement membrane repairing effect. Summary of the invention

[0008] The present invention relates to a rhodiola rosea-based composition comprising a rhodiola rosea supercritical fluid extract, a rhodiola rosea alcohol extract and a rhodiola rosea polysaccharide extract. The rhodiola rosea-based composition of the present invention has the following advantages:

[0009] (1) The prior art has not reported that extracts related to Rhodiola rosea have a skin basement membrane repairing effect. The present invention is the first to discover that the composition based on Rhodiola rosea according to the present invention has a skin basement membrane repairing effect;

[0010] (2) The Rhodiola rosea-based composition of the present invention can play an excellent role in repairing the skin basement membrane at a concentration as low as 0.0056%, and has significant efficacy;

[0011] (3) The Rhodiola rosea-based composition of the present invention uses the natural raw material Rhodiola rosea, and the obtained composition is safe, mild and non-irritating;

[0012] (4) The Rhodiola rosea-based composition of the present invention not only increases the expression of basement membrane components, but also increases the retention of the composition in the skin, thereby increasing its bioavailability, thereby ensuring the efficacy of the active ingredient;

[0013] (5) The present invention has discovered a new barrier repair and anti-aging active combination raw material targeting the skin basement membrane, filling the application gap of Rhodiola rosea cosmetic raw materials in the field of skin basement membrane repair.

[0014] The first aspect of the present invention relates to a composition comprising a supercritical fluid extract of Rhodiola rosea, an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea, wherein the supercritical extract of Rhodiola rosea is prepared by supercritical fluid CO 2 Extracts obtained by extracting Rhodiola rosea raw materials or Rhodiola rosea medicinal material residues that have been extracted with water and / or ethanol. The Rhodiola rosea alcohol extract is obtained by extracting Rhodiola rosea raw materials with ethanol or by extracting Rhodiola rosea raw materials with supercritical fluid CO 2 Extracts obtained from the residues of Rhodiola rosea after extraction and / or water extraction, and Rhodiola rosea polysaccharide extracts are obtained by extracting the Rhodiola rosea raw materials with water or by supercritical fluid CO 2 The extract is obtained by extracting and / or ethanol extracting the polysaccharides in the residue of Rhodiola rosea medicinal material.

[0015] The second aspect of the present invention relates to a supercritical fluid extract of Rhodiola rosea, which is prepared by supercritical fluid CO 2 The extract obtained by extracting Rhodiola rosea raw material or Rhodiola rosea medicinal material residue that has been extracted with water and / or ethanol is prepared by a method comprising the following steps:

[0016] (1) Add the raw material of Rhodiola rosea or the residue of Rhodiola rosea extracted with water and / or ethanol into an extraction kettle, and introduce CO in a supercritical fluid state. 2 performing extraction to obtain a crude extract;

[0017] (2) Add 10-300 times the volume of caprylic acid capric acid triglyceride to the crude extract obtained in step (1), heat to 50-90° C. to dissolve, and then filter to remove impurities.

[0018] The third aspect of the present invention relates to a composition comprising any two components selected from the Rhodiola rosea supercritical fluid extract, Rhodiola rosea alcohol extract and Rhodiola rosea polysaccharide extract defined in the first invention of the present invention.

[0019] The fourth aspect of the present invention relates to the use of the composition according to the first aspect of the present invention, the Rhodiola rosea supercritical fluid extract according to the second aspect of the present invention, the composition according to the third aspect of the present invention, or the Rhodiola rosea supercritical fluid extract, Rhodiola rosea alcohol extract or Rhodiola rosea polysaccharide extract defined in the first aspect of the present invention in the preparation of cosmetics or medicines.

[0020] The fifth aspect of the present invention relates to a cosmetic or a medicine comprising the composition according to the first aspect of the present invention or the Rhodiola rosea supercritical fluid extract according to the second aspect of the present invention or the composition according to the third aspect of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The effects of oil-soluble Rhodiola rosea, Rhodiola rosea, polysaccharide, Rhodiola rosea + polysaccharide and triple-extracted Rhodiola rosea composition according to Example 4 on the gene expression of COL IV protein in HFF-1 cells are shown, wherein *, **, *** and **** respectively indicate p<0.05, p<0.01, p<0.001 and p<0.0001 compared with the control group.

[0022] Figure 2 The effects of oil-soluble Rhodiola rosea, Rhodiola rosea, polysaccharide, Rhodiola rosea + polysaccharide and triple-extracted Rhodiola rosea composition according to Example 4 on the gene expression of Laminin 332 protein in HFF-1 cells are shown, where *, **, *** and **** respectively represent p<0.05, p<0.01, p<0.001 and p<0.0001 compared with the control group.

[0023] Figure 3 The effects of oil-soluble Rhodiola rosea, Rhodiola rosea, polysaccharide, Rhodiola rosea + polysaccharide and triple-extracted Rhodiola rosea composition according to Example 4 on the gene expression of COL VII protein in HFF-1 cells are shown, where *, **, *** and **** respectively indicate p<0.05, p<0.01, p<0.001 and p<0.0001 compared with the control group.

[0024] Figure 4 The effects of oil-soluble Rhodiola rosea, Rhodiola rosea, polysaccharide, Rhodiola rosea + polysaccharide and triple-extracted Rhodiola rosea composition according to Example 4 on the gene expression of COL VII protein in KC cells are shown, wherein *, **, *** and **** respectively indicate p<0.05, p<0.01, p<0.001 and p<0.0001 compared with the control group.

[0025] Figure 5The effects of oil-soluble Rhodiola rosea, Rhodiola rosea, polysaccharide, Rhodiola rosea + polysaccharide and triple-extracted Rhodiola rosea composition according to Example 4 on the gene expression of COL XVII protein in KC cells are shown, wherein *, **, *** and **** respectively indicate p<0.05, p<0.01, p<0.001 and p<0.0001 compared with the control group.

[0026] Figure 6 The effects of oil-soluble Rhodiola rosea, Rhodiola rosea, polysaccharide, Rhodiola rosea + polysaccharide and triple-extracted Rhodiola rosea composition according to Example 4 on the gene expression of HSPG2 in KC cells are shown, wherein *, **, *** and **** respectively indicate p<0.05, p<0.01, p<0.001 and p<0.0001 compared with the control group.

[0027] Figure 7 The retention amounts of the oil-soluble Rhodiola rosea, Rhodiola rosea, polysaccharide and Rhodiola rosea tri-extract composition according to Example 5 in pig skin after 6 h of transdermal absorption are shown, wherein * indicates p<0.05 compared with each single-component group. DETAILED DESCRIPTION OF THE INVENTION

[0029] definition

[0030] As used herein, the terms "s", "min" and "h" stand for "seconds", "minutes" and "hours", respectively.

[0031] As used herein, the term "comprising", which is synonymous with "including", "containing" or "characterized by", is inclusive or open-ended and does not exclude additional, undescribed elements or method steps. However, in the text, each time "comprising" is mentioned, it is intended to cover the alternative embodiments of "consisting essentially of" and "consisting of", wherein "consisting of" excludes any elements or steps not specified, and "consisting essentially of" allows the inclusion of other elements or steps not described that do not materially affect the essential or basic characteristics and novel characteristics of the composition or method in question.

[0032] The term "about" as used herein refers to ±10%, more preferably ±5%, and most preferably ±2% of the value modified by the term, so that a person skilled in the art can clearly determine the scope of the term "about" based on the modified value.

[0033] The terms "triple-extract Rhodiola rosea" and "triple-extract Rhodiola rosea composition" used in the present invention both refer to the composition comprising a Rhodiola rosea supercritical fluid extract, a Rhodiola rosea alcohol extract and a Rhodiola rosea polysaccharide extract according to the first aspect of the present invention.

[0034] When describing the method for preparing Rhodiola rosea polysaccharide extract in the present invention, "optional (3)", "optional (4)" and "optional (5)" respectively mean that step (3), step (4) and step (5) of the method may or may not exist. For example, "optional (3)" means that the method may or may not include step (3).

[0035] I. Three-extract Rhodiola rosea composition

[0036] The first aspect of the present invention relates to a composition comprising a supercritical fluid extract of Rhodiola rosea, an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea, wherein the supercritical extract of Rhodiola rosea is prepared by supercritical fluid CO 2 Extracts obtained by extracting Rhodiola rosea raw materials or Rhodiola rosea medicinal material residues that have been extracted with water and / or ethanol. The Rhodiola rosea alcohol extract is obtained by extracting Rhodiola rosea raw materials with ethanol or by extracting Rhodiola rosea raw materials with supercritical fluid CO 2 Extracts obtained from the residues of Rhodiola rosea after extraction and / or water extraction, and Rhodiola rosea polysaccharide extracts are obtained by extracting the Rhodiola rosea raw materials with water or by supercritical fluid CO 2 The extract is obtained by extracting and / or ethanol extracting the polysaccharides in the residue of Rhodiola rosea medicinal material.

[0037] In some embodiments, the Rhodiola rosea is a plant of the genus Rhodiola, preferably selected from Rhodiola rosea (Rhodiolacrenulata (Hook.f. & Thomson) H.Ohba), Rhodiola calliantha (H.Ohba) H.Ohba), Rhodiola cretinii (Raym.-Hamet) H.Ohba, Rhodiolaheterodonta (Hook.f. & Thomson) Boriss.), Rhodiola kirilowii (Regel) Maxim., Rhodiola linearifolia A.Bor., Rhodiola akirilowii (Regel) Maxim., Rhodiola rosea L., Rhodiolasachalinensis Boriss., Rhodiola serrata H.Ohba) and Rhodiola dumulosa (Franch.) SHFu, and most preferably Rhodiolacrenulata (Hook.f.et Thoms.) H.Ohba.

[0038] In some embodiments, the Rhodiola rosea raw material is a Rhodiola rosea medicinal material. In some embodiments, the Rhodiola rosea raw material is a Rhodiola rosea root medicinal material, a Rhodiola rosea stem medicinal material, or a Rhodiola rosea other part medicinal material. In some embodiments, the Rhodiola rosea raw material is a Rhodiola rosea root medicinal material. In some embodiments, the Rhodiola rosea raw material is first crushed into a 5-30 mesh coarse powder and then extracted.

[0039] In some embodiments, the ratio between the supercritical fluid extract of Rhodiola rosea and the alcohol extract of Rhodiola rosea in the composition is 100:1 to 1:100. In some embodiments, the ratio between the supercritical fluid extract of Rhodiola rosea and the alcohol extract of Rhodiola rosea in the composition is 100:1 to 90:1, 90:1 to 80:1, 80:1 to 70:1, 70:1 to 60:1, 60:1 to 50:1, 50:1 to 40:1, 40:1 to 30:1, 30:1 to 20:1, 20:1 to 10:1, 10:1 to 5:1, 5:1 to or 1:10.

[0040] In some embodiments, the ratio between the alcohol extract of Rhodiola rosea and the polysaccharide extract of Rhodiola rosea in the composition is 1: 1 to 1: 10. In some embodiments, the ratio between the alcohol extract of Rhodiola rosea and the polysaccharide extract of Rhodiola rosea in the composition is 1: 1 to 1: 2, 1: 2 to 1: 3, 1: 3 to 1: 4, 1: 4 to 1: 5, 1: 5 to 1: 6, 1: 6 to 1: 7, 1: 7 to 1: 8, 1: 8 to 1: 9 or 1: 9 to 1: 10.

[0041] I.1 Rhodiola rosea supercritical fluid extract

[0042] In some embodiments, the Rhodiola rosea supercritical fluid extract is prepared by a method comprising the following steps:

[0043] (1) Add the raw material of Rhodiola rosea or the residue of Rhodiola rosea extracted with water and / or ethanol into an extraction kettle, and introduce CO in a supercritical fluid state. 2 performing extraction to obtain a crude extract;

[0044] (2) Add 10-300 times the volume of caprylic acid capric acid triglyceride to the crude extract obtained in step (1), heat to 50-90° C. to dissolve, and then filter to remove impurities.

[0045] In some embodiments, the Rhodiola rosea supercritical fluid extract optionally further comprises caprylic capric triglyceride.

[0046] In some embodiments, the method for preparing a supercritical fluid extract of Rhodiola rosea further comprises the following steps:

[0047] (3) removing (for example, but not limited to, by drying, distillation, solvent extraction, low-temperature crystallization, etc.) the caprylic / capric triglyceride in step (2).

[0048] In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the pressure in the separation kettle I of the extraction device is 5-10 MPa. In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the pressure in the separation kettle I of the extraction device is 5-6 MPa, 6-7 MPa, 7-8 MPa, 8-9 MPa or 9-10 MPa.

[0049] In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the temperature in the separation tank I of the extraction device is 40-60° C. In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the temperature in the separation tank I of the extraction device is 40-45° C., 45-50° C., 50-55° C. or 55-60° C.

[0050] In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the pressure in the separation kettle II of the extraction device is 1-7MPa. In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the pressure in the separation kettle II of the extraction device is 1-2MPa, 2-3MPa, 3-4MPa, 4-5MPa, 5-6MPa or 6-7MPa.

[0051] In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the temperature in the separation tank II of the extraction device is 45-65° C. In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed in an extraction device, wherein the temperature in the separation tank II of the extraction device is 45-50° C., 50-55° C., 55-60° C. or 60-65° C.

[0052] In some embodiments, the pressure in the separation tank I of the extraction apparatus is greater than the pressure in the separation tank II. In some embodiments, the temperature in the separation tank I of the extraction apparatus is less than the temperature in the separation tank II.

[0053] In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed at a pressure of 20-50MPa. In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed at a pressure of 20-25MPa, 25-30MPa, 30-35MPa, 35-40MPa, 40-45MPa or 45-50MPa. In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed at a pressure of 30MPa.

[0054] In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed at a temperature of 40-70° C. In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed at a temperature of 40-45° C., 45-50° C., 50-55° C., 55-60° C., 60-65° C., or 65-70° C. In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed at a temperature of 55° C.

[0055] In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed for 0.5-5h. In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed for 0.5-1h, 1-1.5h, 1.5-2h, 2-2.5h, 2.5-3h, 3-3.5h, 3.5-4h, 4-4.5h or 4.5-5h. In some embodiments, in the step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, supercritical fluid extraction is performed for 2h.

[0056] In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, the supercritical fluid state of CO introduced 2 The flow rate is 10-50 L / h. In some embodiments, in step (1) of the method for preparing a supercritical fluid extract of Rhodiola rosea, the supercritical fluid state of CO introduced 2 The flow rate is 10-15L / h, 15-20L / h, 20-25L / h, 25-30L / h, 30-35L / h, 35-40L / h, 40-45L / h or 45-50L / h.

[0057] In some embodiments, in step (2) of the method for preparing a supercritical fluid extract of Rhodiola rosea, the volume of caprylic capric triglyceride added is 10-300 times the volume of the crude extract obtained in step (1). In some embodiments, in step (2) of the method for preparing a supercritical fluid extract of Rhodiola rosea, the volume of caprylic capric triglyceride added is 10-20 times, 20-30 times, 30-40 times, 40-50 times, 50-60 times, 60-70 times, 70-80 times, 80-90 times, 90-100 times, 100-110 times, 110-120 times, 120-130 times the volume of the crude extract obtained in step (1). In some embodiments, in the step (2) of the method for preparing the supercritical fluid extract of Rhodiola rosea, the volume of caprylic capric triglyceride added is 19 times the volume of the crude extract obtained in step (1). In some embodiments, in step (2) of the method for preparing a supercritical fluid extract of Rhodiola rosea, the volume of caprylic capric triglyceride added is 19-199 times the volume of the crude extract obtained in step (1). In some embodiments, in step (2) of the method for preparing a supercritical fluid extract of Rhodiola rosea, the volume of caprylic capric triglyceride added is 199 times the volume of the crude extract obtained in step (1).

[0058] In some embodiments, in step (2) of the method for preparing a supercritical fluid extract of Rhodiola rosea, 10-300 times the volume of caprylic capric triglyceride is added to the crude extract obtained in step (1), heated to 50-90°C for dissolution, and then filtered to remove impurities. In some embodiments, in step (2) of the method for preparing a supercritical fluid extract of Rhodiola rosea, 10-300 times the volume of caprylic capric triglyceride is added to the crude extract obtained in step (1), heated to 50-55°C, 55-60°C, 60-65°C, 65-70°C, 70-75°C, 75-80°C, 80-85°C or 85-90°C for dissolution, and then filtered to remove impurities.

[0059] I.2 Rhodiola rosea alcohol extract

[0060] In some embodiments, the Rhodiola rosea alcohol extract is prepared by a method comprising an ethanol extraction step, a macroporous resin column purification step, a concentration step, and a drying step.

[0061] In some embodiments, the Rhodiola rosea alcohol extract is prepared by a method comprising the following steps:

[0062] (1) Rhodiola rosea raw material or 2 The Rhodiola rosea medicinal material residue after extraction and / or water extraction is subjected to ultrasonic extraction with 65%-85% ethanol at 20-80° C. for 30-60 minutes, and after the extraction is completed, the Rhodiola rosea raw material residue is filtered to obtain a Rhodiola rosea alcohol extract;

[0063] (2) extracting the Rhodiola rosea raw material residue filtered out in step (1) again with ethanol under the same conditions as in step (1), and performing step (2) 0-3 times;

[0064] (3) if step (2) is performed 0 times, the rhodiola rosea alcohol extract of step (1) is directly concentrated; if step (2) is performed 1-3 times, the rhodiola rosea alcohol extracts of step (1) and step (2) are combined; and the concentrated solution is purified by a macroporous adsorption resin, wherein the macroporous adsorption resin is first eluted with water to remove impurities, and then the macroporous adsorption resin is eluted with 70-80% ethanol to obtain an eluate;

[0065] (4) The eluate obtained in step (3) is concentrated and then dried to obtain the Rhodiola rosea alcohol extract.

[0066] In some embodiments, the Rhodiola rosea alcohol extract is obtained by ethanol extraction, wherein the concentration of the extraction solvent ethanol is 50%-100%. In some embodiments, the concentration of the extraction solvent ethanol is 50%-55%, 55%-60%, 60%-65%, 65%-70%, 70%-75%, 75%-80%, 80%-85%, 85%-90%, 90%-95% or 95%-100%. In some embodiments, the concentration of the extraction solvent ethanol is 65%-85%.

[0067] In some embodiments, the Rhodiola rosea alcohol extract is obtained by ethanol extraction, and the amount of ethanol used for extraction is the same as that of the Rhodiola rosea raw material or the Rhodiola rosea raw material that has been subjected to supercritical fluid CO 2 In some embodiments, the amount of the extraction solvent ethanol used for extraction is 2-50 times the amount of the Rhodiola rosea raw material or the Rhodiola rosea raw material that has been supercritical fluid CO 22-3 times, 3-4 times, 4-5 times, 5-6 times, 6-7 times, 7-8 times, 8-9 times, 9-10 times, 10-11 times, 11-12 times, 12-13 times, 13-14 times, 14-15 times, 15-16 times, 16-17 times, 17-18 times, 18-19 times, 19-20 times, 20-21 times, 21-22 times, 22-23 times, 23-24 times, 24-25 times, 25-26 times, , 26-27 times, 27-28 times, 28-29 times, 29-30 times, 30-31 times, 31-32 times, 32-33 times, 33-34 times, 34-35 times, 35-36 times, 36-37 times, 37-38 times, 38-39 times, 39-40 times, 40-41 times, 41-42 times, 42-43 times, 43-44 times, 44-45 times, 45-46 times, 46-47 times, 47-48 times, 48-49 times or 49-50 times. In some embodiments, the amount of extraction solvent ethanol used for extraction is the amount of Rhodiola rosea raw material or has been extracted with supercritical fluid CO 2 6-10 times the amount of the Rhodiola rosea medicinal material residue after extraction and / or water extraction.

[0068] In some embodiments, the temperature of ultrasonic extraction is 20-80° C. In some embodiments, the temperature of ultrasonic extraction is 20-25° C., 25-30° C., 30-35° C., 35-40° C., 40-45° C., 45-50° C., 50-55° C., 55-60° C., 60-65° C., 65-70° C., 70-75° C., or 75-80° C.

[0069] In some embodiments, the time of ultrasonic extraction is 10-180 minutes. In some embodiments, the time of ultrasonic extraction is 10-20 minutes, 20-30 minutes, 30-40 minutes, 40-50 minutes, 50-60 minutes, 60-70 minutes, 70-80 minutes, 80-90 minutes, 90-100 minutes, 100-110 minutes, 110-120 minutes, 120-130 minutes, 130-140 minutes, 140-150 minutes, 150-160 minutes, 160-170 minutes or 170-180 minutes. In some embodiments, the time of ultrasonic extraction is 30-60 minutes.

[0070] In some embodiments, step (2) of the method for preparing an alcohol extract of Rhodiola rosea is performed 0-3 times. In some embodiments, step (2) of the method for preparing an alcohol extract of Rhodiola rosea is performed 0 times, 1 time, 2 times or 3 times.

[0071] In some embodiments, the macroporous adsorption resin is selected from macroporous adsorption resin AB8, D101, DB-301 and DEAE-52. In some embodiments, the macroporous adsorption resin is macroporous adsorption resin AB8. In some embodiments, the macroporous adsorption resin is D101.

[0072] In some embodiments, the purification by macroporous adsorption resin includes the steps of first eluting the macroporous adsorption resin with water to remove impurities, and then eluting the macroporous adsorption resin with 70-80% ethanol to obtain an eluent. In some embodiments, the amount of water eluent used is 1-5 times the column volume. In some embodiments, the amount of water eluent used is 1-1.5 times the column volume, 1.5-2 times the column volume, 2-2.5 times the column volume, 2.5-3 times the column volume, 3-3.5 times the column volume, 3.5-4 times the column volume, 4-4.5 times the column volume or 4.5-5 times the column volume. In some embodiments, the amount of water eluent used is 1.5-3 times the column volume.

[0073] In some embodiments, the purification by macroporous adsorption resin includes the steps of first eluting the macroporous adsorption resin with water to remove impurities, and then eluting the macroporous adsorption resin with 70-80% ethanol to obtain an eluent. In some embodiments, the amount of ethanol eluent used is 1-10 times the column volume. In some embodiments, the amount of ethanol eluent used is 1-2 times the column volume, 2-3 times the column volume, 3-4 times the column volume, 4-5 times the column volume, 5-6 times the column volume, 6-7 times the column volume, 7-8 times the column volume, 8-9 times the column volume or 9-10 times the column volume. In some embodiments, the amount of ethanol eluent used is 2-5 times the column volume.

[0074] I.3 Rhodiola rosea polysaccharide extract

[0075] In some embodiments, the Rhodiola rosea polysaccharide extract is prepared by a method comprising the following steps:

[0076] (1) Rhodiola rosea raw material or 2 The Rhodiola rosea medicinal material residue after extraction and / or ethanol extraction is added into water, heated under reflux for extraction, and a water extract is obtained;

[0077] (2) filtering the aqueous extract obtained in step (1) and collecting the filtrate, and optionally concentrating the filtrate;

[0078] Optionally (3) adding alcohol to the filtrate obtained in the optionally concentrated step (2), mixing evenly and then standing for a period of time to obtain an alcohol precipitate;

[0079] Optional (4) centrifuging the alcohol precipitate obtained in step (3) to collect the precipitate;

[0080] Optionally (5) adding the precipitate collected in step (4) to water and heating until fully dissolved;

[0081] (6) refrigerating the solution obtained in step (2) or step (5) for a period of time to obtain a suspension;

[0082] (7) Centrifuging and / or filtering the suspension obtained in step (6), collecting the supernatant or filtrate, and optionally concentrating the collected supernatant or filtrate.

[0083] In some embodiments, the method for preparing Rhodiola rosea polysaccharide extract further comprises the following steps:

[0084] (8) Drying the optionally concentrated supernatant or filtrate obtained in step (7).

[0085] I.3.1 Steps (1) of the method for preparing Rhodiola rosea polysaccharide extract

[0086] In some embodiments, in step (1) of the method for preparing a Rhodiola rosea polysaccharide extract, the Rhodiola rosea raw material or the Rhodiola rosea raw material has been subjected to supercritical fluid CO 2 The ratio of the Rhodiola rosea medicinal material residue after extraction and / or ethanol extraction to water is 1:5 to 1:30. In some embodiments, in step (1) of the method for preparing the Rhodiola rosea polysaccharide extract, the Rhodiola rosea raw material or the Rhodiola rosea raw material has been subjected to supercritical fluid CO 2 The ratio of the Rhodiola rosea medicinal material residue to water after extraction and / or ethanol extraction is 1:5 to 1:10, 1:10 to 1:15, 1:15 to 1:20, 1:20 to 1:25 or 1:25 to 1:30. In some embodiments, in step (1) of the method for preparing the Rhodiola rosea polysaccharide extract, the Rhodiola rosea raw material or the Rhodiola rosea raw material that has been subjected to supercritical fluid CO 2 The ratio of the Rhodiola rosea medicinal material residue after extraction and / or ethanol extraction to water is 1:8 to 1:12.

[0087] In some embodiments, in step (1) of the method for preparing a Rhodiola rosea polysaccharide extract, the number of extractions is 1-3 times. In some embodiments, in step (1) of the method for preparing a Rhodiola rosea polysaccharide extract, the number of extractions is 1, 2 or 3 times.

[0088] In some embodiments, in the step (1) of the method for preparing a Rhodiola rosea polysaccharide extract, each extraction is 0.5-5h. In some embodiments, in the step (1) of the method for preparing a Rhodiola rosea polysaccharide extract, each extraction is 0.5-1h, 1-1.5h, 1.5-2h, 2-2.5h, 2.5-3h, 3-3.5h, 3.5-4h, 4-4.5h or 4.5-5h. In some embodiments, in the step (1) of the method for preparing a Rhodiola rosea polysaccharide extract, each extraction is 0.5-3h. In some embodiments, in the step (1) of the method for preparing a Rhodiola rosea polysaccharide extract, each extraction is 1-2h.

[0089] I.3.2 Step (2) of the method for preparing Rhodiola rosea polysaccharide extract

[0090] In some embodiments, in step (2) of the method for preparing a Rhodiola rosea polysaccharide extract, the filtrate is optionally concentrated to 20% to 50% of the original volume of the filtrate.

[0091] In some embodiments, in the step (2) of the method for preparing Rhodiola rosea polysaccharide extract, after concentrating the filtrate, the concentrated filtrate is centrifuged and the supernatant is collected for step (3). In some embodiments, the time of the centrifugation is 5-30 minutes. In some embodiments, the time of the centrifugation is 5-10 minutes, 10-15 minutes, 15-20 minutes, 20-25 minutes or 25-30 minutes. In some embodiments, the time of the centrifugation is 15 minutes. In some embodiments, the centrifugation is centrifugation at a speed of 5000-50000r / min. In some embodiments, the centrifugation is centrifugation at a speed of 5000-10000r / min, 10000-20000r / min, 20000-30000r / min, 30000-40000r / min or 40000-50000r / min. In some embodiments, the centrifugation is centrifugation at a rotation speed of 10000 r / min.

[0092] I.3.3 Step (3) of the method for preparing Rhodiola rosea polysaccharide extract

[0093] In some embodiments, in step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is ethanol or methanol. In some embodiments, in step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is ethanol. In some embodiments, in step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is methanol.

[0094] In some embodiments, in step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the concentration and volume of the added alcohol are such that the concentration of the alcohol in the alcohol precipitate obtained after the addition of alcohol is 40% to 90%. In some embodiments, in step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the concentration and volume of the added alcohol are such that the concentration of the alcohol in the alcohol precipitate obtained after the addition of alcohol is 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80% or 80% to 90%. In some embodiments, in step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the concentration and volume of the added alcohol are such that the concentration of the alcohol in the alcohol precipitate obtained after the addition of alcohol is 50% to 90%.

[0095] In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is 60%-100% alcohol. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is 60%-65% alcohol, 65%-70% alcohol, 70%-75% alcohol, 75%-80% alcohol, 80%-85% alcohol, 85%-90% alcohol, 90%-95% alcohol or 95%-100% alcohol. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is 95% alcohol. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is anhydrous alcohol.

[0096] In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is 60%-100% ethanol. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is 60%-65% ethanol, 65%-70% ethanol, 70%-75% ethanol, 75%-80% ethanol, 80%-85% ethanol, 85%-90% ethanol, 90%-95% ethanol or 95%-100% ethanol. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is 95% ethanol. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol used is anhydrous ethanol.

[0097] In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the volume of the alcohol used is 2-10 times the volume of the liquid obtained in step (2). In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the volume of the alcohol used is 2-3 times, 3-4 times, 4-5 times, 5-6 times, 6-7 times, 7-8 times, 8-9 times or 9-10 times the volume of the liquid obtained in step (2). In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the volume of the alcohol used is 3-8 times the volume of the liquid obtained in step (2). In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, the volume of the alcohol used is 3-5 times the volume of the liquid obtained in step (2).

[0098] In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, after adding alcohol and mixing evenly, let stand for 0.5-36h. In some embodiments, in step (3), after adding alcohol and mixing evenly, let stand for 0.5-3h, 3-6h, 6-9h, 9-12h, 12-15h, 15-18h, 18-21h, 21-24h, 24-27h, 27-30h, 30-33h or 33-36h. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, after adding alcohol and mixing evenly, let stand for 0.5-12h. In some embodiments, in the step (3) of the method for preparing a Rhodiola rosea polysaccharide extract, after adding alcohol and mixing evenly, let stand for 2-8h.

[0099] I.3.4 Step (4) of the method for preparing Rhodiola rosea polysaccharide extract

[0100] In some embodiments, in step (4) of the method for preparing a Rhodiola rosea polysaccharide extract, the alcohol precipitate obtained in step (3) is centrifuged at a speed of 5000-50000 r / min to collect the precipitate. In some embodiments, the alcohol precipitate obtained in step (3) is centrifuged at a speed of 5000-10000 r / min, 10000-15000 r / min, 15000-20000 r / min, 20000-25000 r / min, 25000-30000 r / min, 30000-35000 r / min, 35000-40000 r / min, 40000-45000 r / min or 45000-50000 r / min to collect the precipitate. In some embodiments, the alcohol precipitate obtained in step (3) is centrifuged at a speed of 10000 r / min to collect the precipitate.

[0101] In some embodiments, in step (4) of the method for preparing a Rhodiola rosea polysaccharide extract, the centrifugal time is 5-30 minutes. In some embodiments, the centrifugal time is 5-10 minutes, 10-15 minutes, 15-20 minutes, 20-25 minutes or 25-30 minutes. In some embodiments, the centrifugal time is 10 minutes.

[0102] I.3.5 Step (5) of the method for preparing Rhodiola rosea polysaccharide extract

[0103] In some embodiments, in step (5) of the method for preparing a Rhodiola rosea polysaccharide extract, the amount of water added is an amount of water that can fully dissolve the precipitate collected in step (4) under heating conditions. In some embodiments, in step (5) of the method for preparing a Rhodiola rosea polysaccharide extract, the amount of water added is an amount of water that can make the solid content in the obtained fully dissolved solution be 5%-15%.

[0104] The "solid content" described herein refers to the percentage of the weight of the solid obtained after evaporating the water in a certain weight of the solution to the weight of the solution. For example, if the weight of the solid obtained after evaporating the water in a solution weighing 100g is 10g, the solid content in the solution is 10%.

[0105] In some embodiments, in step (5) of the method for preparing a Rhodiola rosea polysaccharide extract, the amount of water added is 5-50 times the amount of the precipitate collected in step (4). In some embodiments, in step (5) of the method for preparing a Rhodiola rosea polysaccharide extract, the amount of water added is 5-10 times, 10-15 times, 15-20 times, 20-25 times, 25-30 times, 30-35 times, 35-40 times, 40-45 times or 45-50 times the amount of the precipitate collected in step (4).

[0106] In some embodiments, in step (5) of the method for preparing a Rhodiola rosea polysaccharide extract, the precipitate collected in step (4) is heated and dissolved at a temperature of 40-80°C. In some embodiments, in step (5) of the method for preparing a Rhodiola rosea polysaccharide extract, the precipitate collected in step (4) is heated and dissolved at a temperature of 40-45°C, 45-50°C, 50-55°C, 55-60°C, 60-65°C, 65-70°C, 70-75°C or 75-80°C. In some embodiments, in step (5) of the method for preparing a Rhodiola rosea polysaccharide extract, the precipitate collected in step (4) is heated and dissolved at a temperature of 60°C.

[0107] I.3.6 Step (6) of the method for preparing Rhodiola rosea polysaccharide extract

[0108] In some embodiments, in step (6) of the method for preparing a Rhodiola rosea polysaccharide extract, refrigeration is performed at a temperature of 0.1-10° C. In some embodiments, in step (6) of the method for preparing a Rhodiola rosea polysaccharide extract, refrigeration is performed at a temperature of 0.1-1° C., 1-2° C., 2-3° C., 3-4° C., 4-5° C., 5-6° C., 6-7° C., 7-8° C., 8-9° C., or 9-10° C. In some embodiments, in step (6) of the method for preparing a Rhodiola rosea polysaccharide extract, refrigeration is performed at a temperature of 2-8° C.

[0109] In some embodiments, in the step (6) of the method for preparing Rhodiola rosea polysaccharide extract, the refrigeration time is greater than 1h. In some embodiments, the refrigeration time is 1h-3 months. In some embodiments, the refrigeration time is 1-2h, 2-3h, 3-4h, 4-5h, 5-6h, 6-7h, 7-8h, 8-9h, 9-10h, 10-11h, 11-12h, 12h-1 day, 1-2 days, 2-5 days, 5-10 days, 10-15 days, 15-20 days, 20-25 days, 25 days-1 month, 1-2 months, 2-3 months. In some embodiments, the refrigeration time is 2-12h. In some embodiments, the refrigeration time is 4-24h.

[0110] I.3.7 Step (7) of the method for preparing Rhodiola rosea polysaccharide extract

[0111] In some embodiments, in the step (7) of the method for preparing a Rhodiola rosea polysaccharide extract, the suspension obtained in step (6) is centrifuged and filtered. In some embodiments, in the step (7) of the method for preparing a Rhodiola rosea polysaccharide extract, the suspension obtained in step (6) is first centrifuged and then filtered. In some embodiments, in the step (7) of the method for preparing a Rhodiola rosea polysaccharide extract, the suspension obtained in step (6) is first centrifuged, then filtered, and then centrifuged. In some embodiments, in the step (7) of the method for preparing a Rhodiola rosea polysaccharide extract, the suspension obtained in step (6) is centrifuged. In some embodiments, in the step (7) of the method for preparing a Rhodiola rosea polysaccharide extract, the suspension obtained in step (6) is filtered. In some embodiments, the centrifugation and filtration can each be performed 1 time, 2 times, or 3 times.

[0112] I.3.8 Step (8) of the method for preparing Rhodiola rosea polysaccharide extract

[0113] In some embodiments, the drying in step (8) of the method for preparing a Rhodiola rosea polysaccharide extract includes, but is not limited to, freeze drying, vacuum drying, atmospheric pressure drying, spray drying, boiling drying, or a combination thereof.

[0114] II. Rhodiola rosea supercritical fluid extract

[0115] The second aspect of the present invention relates to a supercritical fluid extract of Rhodiola rosea, which is prepared by supercritical fluid CO 2 The extract obtained by extracting Rhodiola rosea raw material or Rhodiola rosea medicinal material residue that has been extracted with water and / or ethanol is prepared by a method comprising the following steps:

[0116] (1) Add the raw material of Rhodiola rosea or the residue of Rhodiola rosea extracted with water and / or ethanol into an extraction kettle, and introduce CO in a supercritical fluid state. 2 performing extraction to obtain a crude extract;

[0117] (2) Add 10-300 times the volume of caprylic acid capric acid triglyceride to the crude extract obtained in step (1), heat to 50-90° C. to dissolve, and then filter to remove impurities.

[0118] The Rhodiola rosea supercritical fluid extract according to the second aspect of the present invention is as defined in detail in Section I.1 above.

[0119] III. Two-extract Rhodiola rosea composition

[0120] The third aspect of the present invention relates to a composition comprising any two components selected from the Rhodiola rosea supercritical fluid extract, Rhodiola rosea alcohol extract and Rhodiola rosea polysaccharide extract defined in the first invention of the present invention.

[0121] In some embodiments, the composition comprises a combination of a supercritical fluid extract of Rhodiola rosea and an alcohol extract of Rhodiola rosea, wherein the ratio between the supercritical fluid extract of Rhodiola rosea and the alcohol extract of Rhodiola rosea is 100:1 to 1:100. In some embodiments, the composition comprises a combination of a supercritical fluid extract of Rhodiola rosea and an alcohol extract of Rhodiola rosea, wherein the ratio between the supercritical fluid extract of Rhodiola rosea and the alcohol extract of Rhodiola rosea is 100:1 to 90:1, 90:1 to 80:1, 80:1 to 70:1, 70:1 to 60:1, 60:1 to 50:1, 50:1 to 40:1, 40:1 to 30:1, 30:1 to 20:1, 20:1 to 10:1, 1:1 to 5:1, 5:1 to 2.5:1, 2.5:1 to 1:1, 1:1 to 1:2.5, 1:2.5 to 1:4, 1:4 to 1:5, 1:5 to 1:10, 1:10 to 1:20, 1:20 to 1:30, 1:30 to 1:40, 1:40 to 1:50, 1:50 to 1:60, 1:60 to 1:70, 1:70 to 1:80, 1:80 to 1:90 or 1:90 to 1:100.

[0122] In some embodiments, the composition comprises a combination of an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea, wherein the ratio between the alcohol extract of Rhodiola rosea and the polysaccharide extract of Rhodiola rosea is 10: 1 to 1: 10. In some embodiments, the composition comprises a combination of an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea, wherein the ratio between the alcohol extract of Rhodiola rosea and the polysaccharide extract of Rhodiola rosea is 10: 1 to 9: 1, 9: 1 to 8: 1, 8: 1 to 7: 1, 7: 1 to 6: 1, 6: 1 to 5: 1, 5: 1 to 4: 1, 4: 1 to 3: 1, 3: 1 to 2: 1, 2: 1 to 1: 1, 1: 1 to 1: 2, 1: 2 to 1: 3, 1: 3 to 1: 4, 1: 4 to 1: 5, 1: 5 to 1: 6, 1: 6 to 1: 7, 1: 7 to 1: 8, 1: 8 to 1: 9 or 1: 9 to 1: 10.

[0123] In some embodiments, the composition comprises a combination of a supercritical fluid extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea, wherein the ratio between the supercritical fluid extract of Rhodiola rosea and the polysaccharide extract of Rhodiola rosea is 100:1 to 1:100. In some embodiments, the composition comprises a combination of a supercritical fluid extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea, wherein the ratio between the supercritical fluid extract of Rhodiola rosea and the polysaccharide extract of Rhodiola rosea is 100:1 to 90:1, 90:1 to 80:1, 80:1 to 70:1, 70:1 to 60:1, 60:1 to 50:1, 50:1 to 40:1, 40:1 to 30:1, 30:1 to 20:1, 20:1 to 10 :1, 10:1 to 5:1, 5:1 to 2.5:1, 2.5:1 to 1:1, 1:1 to 1:2, 1:2 to 1:4, 1:4 to 1:5, 1:5 to 1:10, 1:10 to 1:20, 1:20 to 1:30, 1:30 to 1:40, 1:40 to 1:50, 1:50 to 1:60, 1:60 to 1:70, 1:70 to 1:80, 1:80 to 1:90 or 1:90 to 1:100.

[0124] IV. Use in the preparation of cosmetics or medicines

[0125] The fourth aspect of the present invention relates to the use of the composition according to the first aspect of the present invention, the Rhodiola rosea supercritical fluid extract according to the second aspect of the present invention, the composition according to the third aspect of the present invention, or the Rhodiola rosea supercritical fluid extract, Rhodiola rosea alcohol extract or Rhodiola rosea polysaccharide extract defined in the first aspect of the present invention in the preparation of cosmetics or medicines.

[0126] In some embodiments, the cosmetic or medicine has the effects of repairing the skin basement membrane, anti-wrinkle, anti-aging, epidermal barrier repair, dermal repair, promoting skin wound healing and / or treating blistering skin diseases.

[0127] In some embodiments, the anti-wrinkle, anti-aging, epidermal barrier repair, dermal repair, promotion of skin wound healing and / or treatment of blistering skin diseases effects are brought about by the skin basement membrane repair effect.

[0128] In some embodiments, the cosmetic or medicine has the effects of anti-wrinkle, anti-aging, epidermal barrier repair, dermal repair, promotion of skin wound healing and / or treatment of blistering skin diseases related to skin basement membrane repair.

[0129] In some embodiments, the cosmetics or drugs can promote the gene expression of type IV collagen in human skin fibroblasts (e.g., HFF-1 cells). In some embodiments, the cosmetics or drugs can promote the gene expression of laminin 5 in human skin fibroblasts (e.g., HFF-1 cells). In some embodiments, the cosmetics or drugs can promote the gene expression of type VII collagen in human skin fibroblasts (e.g., HFF-1 cells).

[0130] In some embodiments, the cosmetics or drugs can promote the gene expression of type VII collagen in human epidermal keratinocytes. In some embodiments, the cosmetics or drugs can promote the gene expression of type XVII collagen in human epidermal keratinocytes. The cosmetics or drugs can promote the gene expression of basement membrane protein glycan in human epidermal keratinocytes.

[0131] V. Cosmetics or medicines

[0132] The fifth aspect of the present invention relates to a cosmetic or a medicine comprising the composition according to the first aspect of the present invention or the Rhodiola rosea supercritical fluid extract according to the second aspect of the present invention or the composition according to the third aspect of the present invention.

[0133] In some embodiments, the composition according to the first aspect of the present invention, the supercritical fluid extract of Rhodiola rosea according to the second aspect of the present invention, or the composition according to the third aspect of the present invention accounts for 0.01%-20% by weight of the cosmetic or drug. In some embodiments, the composition according to the first aspect of the present invention, the supercritical fluid extract of Rhodiola rosea according to the second aspect of the present invention, or the composition according to the third aspect of the present invention accounts for 0.01%-0.02%, 0.02%-0.03%, 0.03%-0.04%, 0.04%-0.05%, 0.05%-0.06%, 0.06%-0.07%, 0.07%-0.08% by weight of the drug or cosmetic. %, 0.08%-0.09%, 0.09%-0.1%, 0.1%-1%, 1%-2%, 2%-3%, 3%-4%, 4%-5%, 5%-6%, 6%-7%, 7%-8%, 8%-9%, 9%-10%, 10%-11%, 11%-12%, 12%-13%, 13%-14%, 14%-15%, 15%-16%, 16%-17%, 17%-18%, 18%-19% or 19%-20%.

[0134] In some embodiments, the medicine or cosmetics are used for skin care. In some embodiments, the medicine and cosmetics are used for skin basement membrane repair, anti-wrinkle, anti-aging, epidermal barrier repair, dermal repair, promoting skin wound healing and / or treating blistering skin diseases. In some embodiments, the medicine and cosmetics include the composition according to the first aspect of the present invention or the Rhodiola rosea supercritical fluid extract according to the second aspect of the present invention or the composition according to the third aspect of the present invention and one or more pharmaceutically or cosmetically acceptable carriers, diluents or excipients. In some embodiments, the medicine and cosmetics are creams, emulsions, pastes, ointments, facial masks, gels, lotions or essences. In some embodiments, the medicine and cosmetics of the present invention may also contain one or more other ingredients, such as plant extracts, nutritional additives, surfactants, flavors and fragrances, pigments, preservatives, antioxidants, moisturizers, ultraviolet absorbers, astringents, penetration aids, pH regulators, etc. Those skilled in the art can choose according to their common sense and specific needs.

[0135] In some embodiments, the medicine or cosmetic further comprises one or more other cosmetic active ingredients, such as but not limited to moisturizers, keratin repair agents, oil control agents, oil repellent agents, hydrating agents, anti-allergic agents, anti-aging and anti-wrinkle agents, etc. DETAILED DESCRIPTION

[0136] The present invention can be implemented through the following embodiments, but the present invention is not limited thereto.

[0137] The instruments and equipment used in the embodiments of the present invention are all conventional instruments and equipment in the field and can be replaced by instruments and equipment that meet the corresponding standards.

[0138] The Chinese herbal medicine raw materials used in the embodiments of the present invention are all commercially available unless otherwise specified, and the reagents used in the embodiments of the present invention are all commercially available analytically pure chemical reagents unless otherwise specified.

[0139] The raw materials and instruments and equipment mentioned in the present invention are all commonly used raw materials and instruments and equipment in the field, which are only examples and are not intended to limit the scope of protection of the present invention. Those skilled in the art can select equivalent raw materials and related instruments and equipment based on the disclosure of the present invention.

[0140] Experimental materials, equipment and statistical analysis methods

[0141] Experimental reagents: Rhodiola rosea was obtained from the Rhodiola rosea planting base of Shanghai Xiangyi Bencao Cosmetics Co., Ltd. in Suozhu Township, Tibet; DMEM medium (Gibco, C11995500BT); Australian fetal bovine serum (FBS, SIGMA, F8318); double antibody (P / S, Gibco, 15140122); trypsin containing 0.25% EDTA (Gibco, 25200056); sterile phenol red-free, calcium-magnesium-containing HBSS buffer (Shanghai Maokang Biotechnology Co., Ltd., MS3505); DPBS (Gibco, C14190500BT); PBS (Wuhan Saiweier Biotechnology Co., Ltd., G4202); SteadyPure universal RNA extraction kit (Acry Biotechnology, AG21017); Evo M-MLV reverse transcription reagent premix (for qPCR) (Acry Biotechnology, AG11706); SYBRGreen Pro Taq HS premixed qPCR kit (including Rox) (ACR Biotechnology, AG11718); optical 96-well PCR reaction plate (ABI, 4306737), PCR optical film (ABI, 4360954); salidroside (CAS: 10338-51-9, Shanghai Shidande Standard Technology Service Co., Ltd.); Bama Xiang pig skin (Shanghai Kaikai Technology Trading Co., Ltd.); human skin fibroblasts (Human Skin Fibroblasts, HFF-1) and human epidermal keratinocytes (Human Epidermis Keratinocytes, KC) (purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences).

[0142] Experimental equipment: microplate centrifuge (LABGIC, CF2800M); high-speed refrigerated centrifuge (Eppendorf, Centrifuge 5425R); nucleic acid protein quantifier (Thermofisher, Nanodrop 2000); PCR thermal cycler (ABI, 2720 Thermal Cycler); real-time fluorescence quantitative PCR instrument (ABI, QuantStudio 6&7Flex Real-Time PCR System); SV-12D transdermal diffusion tester (Shanghai Yuyan Scientific Instrument Co., Ltd.), Franz diffusion cell (caliber 20 mm, capacity 7 mL); 1260 Infinity III liquid chromatography system (Agilent Technologies (China) Co., Ltd.).

[0143] Statistical methods: All data are presented as mean ± standard error (SEM) of three independent representatives and are mentioned in the figure legends. Statistical analysis was performed using GraphPad Prism 9 software using unpaired two-tailed Student's t-test and two-way ANOVA with Tukey's multiple comparison analysis to determine significant differences between groups, and values ​​identified as outliers were excluded from statistical analysis. Results were normalized by the mean of the corresponding control group, and P < 0.05 was considered statistically significant.

[0144] Example

[0145] Example 1: Preparation of Rhodiola rosea supercritical fluid extract

[0146] A certain amount of Rhodiola rosea was weighed and loaded into the extraction kettle. While other parameters were unchanged (the pressure of separation kettle I was 6 MPa and the temperature was 50 °C, the pressure of separation kettle II was 4 MPa and the temperature was 55 °C, CO 2 Under the condition of flow control at 25L / h), extracting for 2h at an extraction pressure of 30MPa and an extraction temperature of 55°C obtains a Rhodiola rosea extract, taking out the Rhodiola rosea extract and adding caprylic capric triglyceride (GTCC) thereto to a concentration of 5% of the Rhodiola rosea extract, heating to 60-80°C until the Rhodiola rosea extract is dissolved in the caprylic capric triglyceride, and removing impurities by filtration to obtain a Rhodiola rosea supercritical fluid extract (hereinafter also referred to as "oil-soluble Rhodiola rosea").

[0147] Example 2: Preparation of Rhodiola rosea alcohol extract

[0148] Take out the drug residue in the extraction kettle after supercritical fluid extraction in Example 1, add 65%-85% ethanol in a concentration of 6-10 times the weight of the drug residue, and ultrasonically extract at a temperature of 20-80°C for 30-60 minutes. After filtering, the filtered medicinal residue is extracted for a second time, and the extraction conditions are consistent with the first time, and the filtrates filtered twice are combined to obtain an alcohol extract. After the alcohol extract is vacuum concentrated to a solid content of 10%-15%, ethanol is removed to obtain an alcohol extract concentrate. The alcohol extract concentrate is loaded onto a macroporous resin column AB8 or D101, first eluted with 1.5-3 times the column volume of water to remove impurities, and then eluted with 2-5 times the column volume of 70-80% ethanol to obtain an eluent. The eluent is concentrated and dried to prepare the final dried Rhodiola rosea alcohol extract.

[0149] Example 3: Preparation of Rhodiola rosea polysaccharide extract

[0150] To the medicinal residue after alcohol extraction in Example 2, add 8-12 times the weight of pure water of the medicinal residue, extract at 70-95°C for 1 hour, filter to obtain a crude polysaccharide extract, vacuum concentrate to a solid content of 3%-8%, place the concentrate at 4°C for 4-24 hours, centrifuge to obtain the supernatant, filter to obtain a filtered concentrate, and freeze-dry the filtered concentrate to obtain a Rhodiola rosea polysaccharide extract.

[0151] Example 4: Efficacy verification (detection of gene expression of main components of skin basement membrane (DEJ-BM))

[0152] 1. Experimental Methods

[0153] (1) Cell culture

[0154] The cells used in the experiment were human skin fibroblasts (HFF-1) and human epidermal keratinocytes (KC) from the cell bank of the Chinese Academy of Sciences in Shanghai. HFF-1 cells were cultured in DMEM medium (containing 15% fetal bovine serum (FBS), 1% penicillin / streptomycin (1% p / s)), and KC cells were cultured in EpiLife TM Culture medium (containing human keratinocyte growth supplement (HKGS), 1% penicillin / streptomycin (1% p / s)), and both cells were cultured at 37°C and 5% CO 2 The cells were cultured in an incubator at 4 °C. After that, fresh culture medium was replaced every 2-3 days. When the cells reached about 80% confluence, 0.25% trypsin-EDTA was used for cell digestion, and the cells were counted using an automatic cell counter. The cells were passaged and the 5th to 10th passages were used for the following experiments.

[0155] (2) Cell administration treatment

[0156] After reaching 90% confluence, the cells were digested with trypsin and seeded in 24-well cell culture plates at 1 × 10 5 cells and cultured for 24 hours. When the cell confluence reached 80%, single-component samples (supercritical fluid extract of Rhodiola rosea, alcohol extract of Rhodiola rosea and polysaccharide extract of Rhodiola rosea), two-component combination samples (combination of two components of alcohol extract of Rhodiola rosea and polysaccharide extract of Rhodiola rosea) and three-component combination samples (combination of three components of supercritical fluid extract of Rhodiola rosea, alcohol extract of Rhodiola rosea and polysaccharide extract of Rhodiola rosea, also known as "three-extract Rhodiola rosea composition") were then administered. The drug applied to HFF-1 cells was prepared using serum-free DMEM, and the drug applied to KC cells was prepared using EpiLife TM The culture medium was prepared, and the control group was a culture medium without the sample to be tested, and cultured for 24 hours at a temperature of 37°C and a 5% carbon dioxide atmosphere. The drug preparation concentration table is shown in Table 1 below:

[0157] Table 1

[0158]

[0159] (3) Cell lysis and total RNA collection

[0160] After the cell culture after drug treatment is completed, the culture medium is completely removed, 1 mL of DPBS is added to wash the cells, 100 μL of cell lysis solution is added, and the cells are fully lysed by pipetting for 5 minutes. The cell lysate is transferred to a centrifuge tube, vortexed and mixed, centrifuged at 12000 rpm for 2 minutes, and the supernatant is taken for the following operations.

[0161] Add the collected supernatant to the genomic DNA removal column, centrifuge at 12000rpm for 30sec, and retain the filtrate. Slowly add 70% ethanol (1 times the volume of the supernatant) to the filtrate, mix well, transfer the resulting solution and precipitate to the RNase-Free adsorption column, centrifuge at 12000rpm for 30sec, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube.

[0162] (4) RNA purification and concentration determination

[0163] Add 700 μL of deproteinization solution to the RNase-Free adsorption column, centrifuge at 12000 rpm for 30 seconds, discard the waste liquid, and return the adsorption column to the collection tube. Add 500 μL of rinse solution to the RNase-Free adsorption column, let it stand at room temperature for 2 minutes, centrifuge at 12000 rpm for 60 seconds, discard the waste liquid, and return the adsorption column to the collection tube. Repeat the wash, centrifuge at 12000 rpm for 2 minutes, and pour out the waste liquid. Leave the RNase-Free adsorption column at room temperature for 2 minutes to thoroughly dry the residual rinse solution in the adsorption material.

[0164] Transfer the RNase-Free adsorption column into a new RNase-Free centrifuge tube and drop 50 μL RNase-Free ddH 2 O, placed at room temperature for 2 min, centrifuged at 12000 rpm for 2 min to obtain RNA solution. The RNA concentration in the sample was determined using NanoDrope2000.

[0165] (5) RNA reverse transcription and cDNA first-chain synthesis

[0166] cDNA synthesis was performed according to the total RNA content of 1000 ng, a certain volume of RNA sample was taken, and reverse transcription was performed according to the 20 μL system.

[0167] (6) RT-qPCR detection of gene expression levels

[0168] Use RNase-free ddHO 2 After diluting the cDNA obtained by reverse transcription 10 times, qPCR reaction was prepared according to the system of 20 μL per well = 2 μL sample cDNA template + 7.2 μL water + 0.4 μL primer (forward) + 0.4 μL (reverse) + 10 μL SYBR Green premixed dye to detect the transcription level of each skin basement membrane component. The primer sequences were designed as follows:

[0169] Table 2

[0170]

[0171]

[0172] (7) Statistical analysis

[0173] According to 2 -(ΔΔCT) The relative expression of each gene was calculated by the PCR method, and GAPDH was selected as the housekeeping gene to standardize the gene expression of the test samples. All data are presented as the mean ± standard error (SEM) of three independent representatives and are mentioned in the legends. Statistical analysis was performed using GraphPad Prism 9 software, using unpaired two-tailed Student's t-test and two-way ANOVA with Tukey's multiple comparison analysis to determine significant differences between groups, and values ​​identified as outliers were excluded from the statistical analysis. The results were normalized by the mean of the corresponding control group, and P < 0.05 was considered statistically significant.

[0174] 2. Results and Discussion

[0175] The test results above are as follows Figure 1-6As shown in Table 3 below:

[0176] Table 3

[0177]

[0178] Depend on Figure 1-6 As can be seen from the results in Table 3, for human dermal fibroblasts, a single component of 0.005% oil-soluble Rhodiola rosea can increase the gene expression of laminin 5 (Laminin-332) and type VII collagen (COL VII) in HFF-1 cells, with gene upregulation rates of 81.32% and 30.62%, respectively. The composite double component of 0.0005% Jiangxue + 0.01% polysaccharide has no obvious promoting effect on the HFF-1 basement membrane components. Tri-extracted Rhodiola rosea can significantly promote the gene expression of type IV collagen (COLIV), COL VII and Laminin-332 in HFF-1 cells, with upregulation rates of 22.94%, 69.51% and 30.20%, respectively. Compared with single / double components, Tri-extracted Rhodiola rosea has a synergistic effect on the gene expression of COL VII and COL IV.

[0179] For human epidermal keratinocytes, single component 0.005% oil-soluble Rhodiola rosea can promote the gene expression of COL VII, type XVII collagen (COL XVII) and basement membrane proteoglycan (HSPG2), with upregulation rates of 65%, 31.64% and 68.92%, respectively. The composite double component 0.0001% Jiangxue + 0.0005% polysaccharide can significantly promote the gene expression of COLVII and HSPG2, with growth rates of 65.91% and 171.64%, respectively. Tri-extracted Rhodiola rosea can significantly upregulate the expression of COL VII, COL XVII and HSPG2, with upregulation rates of 155.77%, 28.92% and 251.77%, respectively. Compared with single / double components, the three components have a synergistic effect on the gene expression of COL VII and HSPG2.

[0180] 3. Experimental conclusion

[0181] The experimental results of two human skin cells showed that after the cells were cultured with the addition of Rhodiola rosea, the gene expression of the skin basement membrane components COL IV, COL VII, COL XVII, HSPG2 and Laminin-332 was significantly improved, so the Rhodiola rosea combination has the effect of repairing the skin basement membrane. Compared with single and double components, Rhodiola rosea also has a significant synergistic effect on the expression of COLVII and HSPG2 genes.

[0182] Numerous studies have reported that most DEJ-BM components are altered during aging. Immunolabeling analysis showed decreased expression levels of laminin-332, collagen IV, collagen VII, collagen XVII, and collagen XVIII in the skin of the elderly. The expression of epidermal perlecan was significantly decreased at both the protein and mRNA levels during skin aging. The expression of collagen XVII is greatly reduced during aging, resulting in hemidesmosome changes, detachment and upward migration of keratinocytes from the DEJ-BM, and decreased elasticity and strength of human skin. In addition to its important role in maintaining dermal-epidermal cohesion, collagen XVII also plays a role in interfollicular keratinocyte proliferation and hair follicle morphogenesis. Colocalization of elastic fibers and type VII collagen can maintain skin structure, thereby preventing wrinkles and sagging, all of which may be affected by the deficiency of DEJ-BM components during aging.

[0183] In short, changes in the composition and structure of the ECM of the skin basement membrane gradually change the mechanical integrity of the DEJ-BM, resulting in impaired responses of cells and tissues to mechanical forces, which in turn leads to impaired skin barrier function. Therefore, the skin basement membrane repair composition of the present invention can significantly promote the expression of DEJ-BM components under the condition of adding at a lower concentration, can play a role in repairing the basement membrane, is highly safe, mild and non-irritating, and is thus beneficial to the repair of the skin barrier and improves skin wrinkles and functional disorders caused by aging.

[0184] Example 5: Evaluation of skin permeation and absorption performance

[0185] 1. Experimental method: Testing the in vitro permeability and absorption capacity of Rhodiola rosea in pig skin

[0186] The skin barrier is very important for skin health, but the existence of the skin barrier will also affect the effect of topical preparations and the penetration and absorption of active substances. In order to verify whether the active substances can enter the skin to exert their effects, the consideration of the transdermal permeation of raw materials is very critical. The Franz diffusion cell permeation test method was used for evaluation, and the test sample inside the pig skin was collected. The content of the known test sample marker salidroside was detected using a high-performance liquid chromatography (HPLC) to determine whether the three-extracted Rhodiola rosea has good permeability and can act on the basement membrane.

[0187] (1) Pigskin and device preparation

[0188] Take the pretreated fresh pig skin (pig skin isolated within 24 hours), soak it in physiological saline for about 10 minutes, take it out, absorb the moisture on the surface of the pig skin with filter paper, cut off the excess part of the pig skin edge according to the shape of the diffusion cell, and fix the pig skin on the Franz diffusion cell.

[0189] (2) Transdermal penetration of the test substance

[0190] Place the matching magnet in the receiving pool, and fix the pig skin between the supply pool and the receiving pool. After fixation, fill the receiving pool with physiological saline, exhaust the air, and make the bottom surface of the pig skin in close contact with the receiving solution. Spread 1mL of the test sample evenly on the pig skin (the sample solvent is used as a blank control), place the diffusion cell in the transdermal diffusion tester, set the instrument parameters: 32±0.5℃, 400rpm / min, and the transdermal duration is 6h; then, rinse the pig skin surface sample with physiological saline, absorb the surface moisture with filter paper, cut off the edge of the pig skin, retain the pig skin involved in the transdermal part, cut the skin into rice grain size or homogenate with surgical scissors, and place it in 2mL of physiological saline for 24 hours at 4℃. After the extraction is completed, centrifuge at 10000rpm for 10min, and the supernatant obtained is the "skin extract".

[0191] (3) Quantitative detection of salidroside

[0192] A series of concentrations of reference substance (Salidroside) was used for HPLC quantitative detection. The reference substance was accurately weighed (measured) to prepare a series of concentrations of reference substance solution; a certain amount of the series of concentrations of reference substance solution and the recovered solution of the test sample of each test group were accurately taken, sampled, and the liquid chromatogram was recorded. The peak area of ​​salidroside in the reference substance solution and the recovered solution of the test sample of the test group was measured, and the content of salidroside in each test sample was calculated.

[0193] (4) Data analysis and calculation

[0194] The results of the "skin extract" showed statistical differences, indicating that the test product can be absorbed by the skin, so the retention amount of salidroside in the skin was calculated:

[0195] Intradermal retention (μg / cm 2 )=c·V / S

[0196] c is the concentration of active substance in the pig skin extract, in μg / mL; V is the total volume of the extract, in mL; S is the transdermal area of ​​pig skin, in cm 2 .

[0197] 2. Results and Discussion

[0198] This experiment uses salidroside as the detection index of each component and its combination, and observes the transdermal effect of three-extracted Rhodiola rosea by comparing the retention amount of salidroside in pig skin. The experimental results are as follows Figure 7 shown.

[0199] Depend on Figure 7The results of the skin retention showed that no salidroside was detected in the skin after 6 hours of transdermal administration of 5% supercritical fluid extract of Rhodiola rosea (oil-soluble Rhodiola rosea), and the retention of 1% alcohol extract of Rhodiola rosea (Jiangxue) in the skin was 11.625ug / cm 2 The retention rate of 5% Rhodiola rosea polysaccharide extract in the skin is 4.09ug / cm 2 The retention rate of the triple-extracted Rhodiola rosea, which contains 0.005% Rhodiola rosea supercritical fluid extract (oil-soluble Rhodiola rosea), 0.0001% Rhodiola rosea alcohol extract (Jiangxue) and 0.0005% Rhodiola rosea polysaccharide extract, in the skin is 22.392ug / cm 2 .

[0200] Compared with the single component, the retention of salidroside in the skin of the three-extracted Rhodiola rosea was significantly increased after 6 hours of transdermal perfusion. The theoretical retention of the combination of 1% Jiangxue + 5% polysaccharide was 15.715ug / cm 2 The retention rate of the three components (0.0001% Jiangxue + 0.0005% polysaccharide + 0.005% oil-soluble rhodiola) in the three-extracted rhodiola rosea skin reached 22.392ug / cm 2 . After 6 hours of transdermal application of oil-soluble rhodiola rosea, no salidroside was detected in the skin, which is speculated to be due to the low content of salidroside in oil-soluble rhodiola rosea. It is speculated that oil-soluble rhodiola rosea may penetrate into the subcutaneous tissue more easily, thus effectively increasing the retention of salidroside in the single-component jiangxue and polysaccharide in the skin, thereby significantly improving the bioavailability of three-extracted rhodiola rosea after application to the skin. Therefore, three-extracted rhodiola rosea can act on the basement membrane of the skin and exert barrier repair and anti-wrinkle effects.

[0201] 3. Experimental conclusion

[0202] According to the results of in vitro pig skin permeation and absorption of Rhodiola rosea, the Rhodiola rosea combination can better act on the skin basement membrane when it stays in the skin, so as to improve the bioavailability of the effective active ingredients in the product. The transdermal experimental data provides a scientific basis for the application of the combination in cosmetics.

Claims

1. A composition comprising a supercritical fluid extract of Rhodiola rosea, an alcohol extract of Rhodiola rosea and a polysaccharide extract of Rhodiola rosea, wherein the supercritical extract of Rhodiola rosea is an extract obtained by extracting a Rhodiola rosea raw material or Rhodiola rosea medicinal material residue after water extraction and / or ethanol extraction with supercritical fluid CO2, the alcohol extract of Rhodiola rosea is an extract obtained by extracting a Rhodiola rosea raw material or Rhodiola rosea medicinal material residue after supercritical fluid CO2 extraction and / or water extraction with ethanol, and the polysaccharide extract of Rhodiola rosea is an extract obtained by extracting polysaccharides in the Rhodiola rosea raw material or Rhodiola rosea medicinal material residue after supercritical fluid CO2 extraction and / or ethanol extraction with water.

2. The composition according to claim 1, wherein the Rhodiola rosea is a plant of the genus Rhodiola, preferably selected from Rhodiola crenulata (Hook.f. & Thomson) H.Ohba, Rhodiola calliantha (H.Ohba) H.Ohba, Rhodiola cretinii (Raym.-Hamet) H.Ohba, Rhodiola heterodonta (Hook.f. & Thomson) Boriss., Rhodiola kirilowii (Regel) Maxim., Rhodiola linearifolia A.Bor., Rhodiola akirilowii (Regel) Maxim., Rhodiola rosea L., Rhodiola sachalinensis Boriss., Rhodiola serrata H.Ohba) and Rhodiola dumulosa (Franch.) SHFu, and most preferably Rhodiolacrenulata (Hook.f.et Thoms.) H.Ohba.

3. The composition according to claim 1 or 2, wherein the Rhodiola rosea supercritical fluid extract is prepared by a method comprising the following steps: (1) adding the raw material of Rhodiola rosea or the residue of Rhodiola rosea after water extraction and / or ethanol extraction into an extraction kettle, and introducing CO2 in a supercritical fluid state to extract to obtain a crude extract; (2) Add 10-300 times the volume of caprylic acid capric acid triglyceride to the crude extract obtained in step (1), heat to 50-90° C. to dissolve, and then filter to remove impurities.

4. The composition according to any one of the preceding claims, wherein the Rhodiola rosea alcohol extract is prepared by a method comprising an ethanol extraction step, a macroporous resin column purification step, a concentration step, and a drying step.

5. The composition according to any one of the preceding claims, wherein the Rhodiola rosea polysaccharide extract is prepared by a method comprising the following steps: (1) adding Rhodiola rosea raw material or Rhodiola rosea medicinal material residue extracted with supercritical fluid CO2 and / or ethanol to water, heating and refluxing for extraction, and obtaining a water extract; (2) filtering the aqueous extract obtained in step (1) and collecting the filtrate, and optionally concentrating the filtrate; Optionally (3) adding alcohol to the filtrate obtained in the optionally concentrated step (2), mixing evenly and then standing for a period of time to obtain an alcohol precipitate; Optional (4) centrifuging the alcohol precipitate obtained in step (3) to collect the precipitate; Optionally (5) adding the precipitate collected in step (4) to water and heating until fully dissolved; (6) refrigerating the solution obtained in step (2) or step (5) for a period of time to obtain a suspension; (7) Centrifuging and / or filtering the suspension obtained in step (6), collecting the supernatant or filtrate, and optionally concentrating the collected supernatant or filtrate.

6. The composition according to any one of the preceding claims, wherein the ratio between the Rhodiola rosea supercritical fluid extract and the Rhodiola rosea alcohol extract is 100:1 to 1:100, and the ratio between the Rhodiola rosea alcohol extract and the Rhodiola rosea polysaccharide extract is 1:1 to 1:

10.

7. A supercritical fluid extract of Rhodiola rosea, which is an extract obtained by extracting Rhodiola rosea raw materials or Rhodiola rosea medicinal material residues extracted with water and / or ethanol by supercritical fluid CO2, and the supercritical fluid extract of Rhodiola rosea is prepared by a method comprising the following steps: (1) adding the raw material of Rhodiola rosea or the residue of Rhodiola rosea after water extraction and / or ethanol extraction into an extraction kettle, and introducing CO2 in a supercritical fluid state to extract to obtain a crude extract; (2) Add 10-300 times the volume of caprylic acid capric acid triglyceride to the crude extract obtained in step (1), heat to 50-90° C. to dissolve, and then filter to remove impurities.

8. The Rhodiola rosea supercritical fluid extract according to claim 7, wherein in step (1), the pressure during supercritical fluid extraction is 20-50 MPa, the temperature is 40-70°C, and the extraction time is 0.5-5h.

9. A composition comprising any two components selected from the group consisting of the Rhodiola rosea supercritical fluid extract, the Rhodiola rosea alcohol extract and the Rhodiola rosea polysaccharide extract as defined in any one of claims 1 to 8.

10. Use of the composition according to any one of claims 1-6, the Rhodiola rosea supercritical fluid extract according to claim 7 or 8, the composition according to claim 9, or an extract selected from any one of the Rhodiola rosea supercritical fluid extract, the Rhodiola rosea alcohol extract and the Rhodiola rosea polysaccharide extract defined in any one of claims 1-8 in the preparation of cosmetics or medicines.

11. The use according to claim 10, wherein the cosmetic or medicine has the effects of repairing skin basement membrane, anti-wrinkle, anti-aging, epidermal barrier repair, dermal repair, promoting skin wound healing and / or treating blistering skin diseases.

12. A cosmetic or a medicine comprising the composition according to any one of claims 1 to 6, or the Rhodiola rosea supercritical fluid extract according to claim 7 or 8, or the composition according to claim 9.

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