An improved rhodiola rosea polysaccharide extract, preparation method and use thereof

Abstract

The present invention provides an improved method for preparing a Rhodiola rosea polysaccharide extract, wherein the method is characterized in that a step of removing the precipitate after refrigeration is added after the traditional water extraction and alcohol precipitation step for polysaccharide extraction. The present invention also provides a Rhodiola rosea polysaccharide extract prepared by the improved method for preparing a Rhodiola rosea polysaccharide extract. The present invention also relates to the use of the Rhodiola rosea polysaccharide extract in the preparation of cosmetics or medicines, and the present invention further relates to cosmetics or medicines containing the Rhodiola rosea polysaccharide extract.

Description

Technical Field

[0001] The present invention relates to a method for preparing an improved Rhodiola rosea polysaccharide extract. The present invention also relates to a Rhodiola rosea polysaccharide extract prepared by the improved method for preparing the Rhodiola rosea polysaccharide extract, use of the Rhodiola rosea polysaccharide extract in preparing cosmetics or medicines, and cosmetics or medicines containing the Rhodiola rosea polysaccharide extract. Background Art

[0002] There are 73 species, 2 subspecies and 7 varieties of Rhodiola rosea in the Crassulaceae family recorded in the Flora of China. They are mainly distributed in the high-altitude and pollution-free areas at an altitude of 3500-5000 meters in the northern hemisphere. Among them, Rhodiola rosea is included in the 2020 edition of the "Chinese Pharmacopoeia". It has the effects of invigorating qi and activating blood circulation, unblocking meridians and relieving asthma. It can be used to treat symptoms such as qi deficiency and blood stasis, chest pain and heart pain, hemiplegia due to stroke, fatigue and asthma.

[0003] So far, more than 180 compounds have been found in Rhodiola plants, mainly phenoxyethanol, flavonoids, terpenes, lignin aliphatic compounds, etc. The representative chemical components are salidroside and tyrosol, which have strong antioxidant effects and are considered to be the main active ingredients and indicator ingredients in Rhodiola. Polysaccharide is one of the main active ingredients in Rhodiola. In recent years, the research on Rhodiola polysaccharide has gradually increased. Modern pharmacological research shows that Rhodiola polysaccharide has multiple effects such as antioxidant, anti-aging, and anti-radiation. It not only has high medicinal value, but also has high application value in cosmetics and health foods.

[0004] However, Rhodiola rosea polysaccharide extracts are prone to precipitation during storage and application, which has some adverse effects on their application in cosmetics: first, precipitation may affect the stability and appearance of the product, causing consumers to have doubts about the quality of the product; second, precipitation may also have a negative impact on the uniformity and shelf life of the product, which may limit its use in cosmetic formulations; in addition, the presence of precipitation may affect the biological activity of Rhodiola rosea polysaccharide extracts, thereby reducing their efficacy in cosmetics.

[0005] Therefore, there is an urgent need for a simple and efficient preparation method that can effectively solve the problem that Rhodiola rosea polysaccharide extracts are prone to precipitation during storage and application. At the same time, there is also an urgent need for a Rhodiola rosea polysaccharide extract that is not prone to precipitation during long-term storage and application, and has reduced cytotoxicity and improved desired cosmetic effects (such as anti-oxidation, anti-aging, anti-inflammatory, repair, etc.). Summary of the invention

[0006] The present invention relates to an improved method for preparing a Rhodiola rosea polysaccharide extract, wherein the method is characterized in that a step of removing the precipitate after refrigeration is added after the traditional water extraction and alcohol precipitation step for polysaccharide extraction. The improved method for extracting Rhodiola rosea polysaccharide has the following advantages:

[0007] (1) The extraction method of Rhodiola rosea polysaccharide provided by the present invention has a simple process, and the equipment used is commonly used in industry and can be directly used in industrial production;

[0008] (2) The inventors of the present invention unexpectedly discovered that the solubility and stability of the Rhodiola rosea polysaccharide extract prepared by the method for preparing the Rhodiola rosea polysaccharide extract of the present invention, characterized in that a step of removing the precipitate after refrigeration is added after the traditional water extraction and alcohol precipitation step, is significantly improved;

[0009] (3) The cytotoxicity of the Rhodiola rosea polysaccharide extract prepared by the method for preparing the Rhodiola rosea polysaccharide extract of the present invention is significantly reduced;

[0010] (4) The Rhodiola rosea polysaccharide extract prepared by the preparation method of the Rhodiola rosea polysaccharide extract of the present invention can significantly promote the expression of fibroblast collagen genes and epidermal cell barrier genes, and has significantly improved efficacy in anti-aging and repair;

[0011] (5) The Rhodiola rosea polysaccharide extract prepared by the method for preparing the Rhodiola rosea polysaccharide extract of the present invention has significantly improved efficacy in anti-inflammatory and antioxidant aspects;

[0012] (6) The present invention is the first to discover that Rhodiola rosea polysaccharide extract has significant activity in resisting photodamage and repairing.

[0013] The first aspect of the present invention relates to a method for preparing a Rhodiola rosea polysaccharide extract, the method comprising the following steps:

[0014] (1) adding the Rhodiola rosea raw material into water, heating and refluxing for extraction, and obtaining a water extract;

[0015] (2) filtering the aqueous extract obtained in step (1) and collecting the filtrate, and optionally concentrating the filtrate;

[0016] (3) adding alcohol to the filtrate obtained in the optionally concentrated step (2), mixing evenly and letting stand for a period of time to obtain an alcohol precipitate;

[0017] (4) centrifuging the alcohol precipitate obtained in step (3) and collecting the precipitate;

[0018] (5) adding the precipitate collected in step (4) into water and heating until fully dissolved;

[0019] (6) refrigerating the solution obtained in step (5) for a period of time to obtain a suspension;

[0020] (7) Centrifuging and / or filtering the suspension obtained in step (6), collecting the supernatant or filtrate, and optionally concentrating the collected supernatant or filtrate.

[0021] The second aspect of the present invention relates to a Rhodiola rosea polysaccharide extract, which is prepared by the method described in the first aspect of the present invention.

[0022] The third aspect of the present invention relates to use of the Rhodiola rosea polysaccharide extract according to the second aspect of the present invention in the preparation of cosmetics or medicines.

[0023] The fourth aspect of the present invention relates to a cosmetic or a medicine comprising the Rhodiola rosea polysaccharide extract according to the second aspect of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The effects of Examples 1-3 and Comparative Examples 1-4 on HaCaT cell viability were detected by Hoechst staining of cell nuclei.

[0025] Figure 2 The effects of Examples 1-3 and Comparative Examples 1-4 on the production of ROS by HaCaT cells induced by H2O2 are shown.

[0026] Figure 3 The effects of Examples 1-3 and Comparative Examples 1-4 on the viability of RAW264.7 cells are shown.

[0027] Figure 4 The effects of Examples 1-3 and Comparative Examples 1-4 on the LPS-induced IL-6 secretion of RAW264.7 cells are shown.

[0028] Figure 5 The effects of Examples 1-3 and Comparative Examples 1-4 on the expression of type III collagen gene in HFF-1 cells induced by UVA are shown.

[0029] Figure 6 The effects of Examples 1-3 and Comparative Examples 1-4 on the expression of CLDN1 gene in UVA-induced HaCaT cells are shown. DETAILED DESCRIPTION OF THE INVENTION

[0031] definition

[0032] As used herein, the terms "s", "min" and "h" stand for "seconds", "minutes" and "hours", respectively.

[0033] 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.

[0034] 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.

[0035] Preparation method of Rhodiola rosea polysaccharide extract

[0036] The first aspect of the present invention relates to a method for preparing a Rhodiola rosea polysaccharide extract, the method comprising the following steps:

[0037] (1) adding the Rhodiola rosea raw material into water, heating and refluxing for extraction, and obtaining a water extract;

[0038] (2) filtering the aqueous extract obtained in step (1) and collecting the filtrate, and optionally concentrating the filtrate;

[0039] (3) adding alcohol to the filtrate obtained in the optionally concentrated step (2), mixing evenly and letting stand for a period of time to obtain an alcohol precipitate;

[0040] (4) centrifuging the alcohol precipitate obtained in step (3) and collecting the precipitate;

[0041] (5) adding the precipitate collected in step (4) into water and heating until fully dissolved;

[0042] (6) refrigerating the solution obtained in step (5) for a period of time to obtain a suspension;

[0043] (7) Centrifuging and / or filtering the suspension obtained in step (6), collecting the supernatant or filtrate, and optionally concentrating the collected supernatant or filtrate.

[0044] In some embodiments, the method further comprises the following steps:

[0045] (8) Drying the optionally concentrated supernatant or filtrate obtained in step (7).

[0046] In some embodiments, the Rhodiola rosea is a plant of the genus Rhodiola, preferably one or more selected from Rhodiolacrenulata (Hook.f.et Thoms.) H.Ohba, Rhodiola rosea L., Rhodiola sacra (Prain ex Raym.-Hamet) SHFu, Rhodiola fastigiata (Hook.f.&Thomson) SHFu, and Rhodiola dumulosa (Franch.) SHFu, most preferably Rhodiola crenulata (Hook.f.et Thoms.) H.Ohba.

[0047] Step (1)

[0048] 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. In some embodiments, the Rhodiola rosea raw material is first crushed into a 5-30 mesh coarse powder and then extracted.

[0049] In some embodiments, in step (1), the ratio of Rhodiola rosea raw material to water is 1:5 to 1:30. In some embodiments, in step (1), the ratio of Rhodiola rosea raw material to water 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), the ratio of Rhodiola rosea raw material to water is 1:10.

[0050] In some embodiments, in step (1), the number of extractions is 1-3 times. In some embodiments, in step (1), the number of extractions is 1, 2 or 3 times.

[0051] In some embodiments, in step (1), each extraction is 0.5-5h. In some embodiments, in step (1), 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 step (1), each extraction is 0.5-3h. In some embodiments, in step (1), each extraction is 1-2h.

[0052] Step (2)

[0053] In some embodiments, in step (2), the filtrate is optionally concentrated to 20% to 50% of the original volume of the filtrate.

[0054] In some embodiments, in step (2), 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 speed of 10000r / min.

[0055] Step (3)

[0056] In some embodiments, in step (3), the alcohol used is ethanol or methanol. In some embodiments, in step (3), the alcohol used is ethanol. In some embodiments, in step (3), the alcohol used is methanol.

[0057] In some embodiments, in step (3), the concentration and volume of the added alcohol are such that the concentration of the alcohol in the alcohol precipitation solution obtained after the addition of alcohol is 40% to 90%. In some embodiments, in step (3), the concentration and volume of the added alcohol are such that the concentration of the alcohol in the alcohol precipitation solution 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), the concentration and volume of the added alcohol are such that the concentration of the alcohol in the alcohol precipitation solution obtained after the addition of alcohol is 50% to 90%.

[0058] In some embodiments, in step (3), the alcohol used is 60%-100% alcohol. In some embodiments, in step (3), 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 step (3), the alcohol used is 95% alcohol. In some embodiments, in step (3), the alcohol used is anhydrous alcohol.

[0059] In some embodiments, in step (3), the alcohol used is 60%-100% ethanol. In some embodiments, in step (3), 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 step (3), the alcohol used is 95% ethanol. In some embodiments, in step (3), the alcohol used is anhydrous ethanol.

[0060] In some embodiments, in step (3), the volume of the alcohol used is 2-10 times the volume of the liquid obtained in step (2). In some embodiments, in step (3), 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 step (3), the volume of the alcohol used is 3-8 times the volume of the liquid obtained in step (2). In some embodiments, in step (3), the volume of the alcohol used is 3-5 times the volume of the liquid obtained in step (2).

[0061] In some embodiments, in step (3), 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 step (3), after adding alcohol and mixing evenly, let stand for 0.5-12h. In some embodiments, in step (3), after adding alcohol and mixing evenly, let stand for 2-8h.

[0062] Step (4)

[0063] In some embodiments, 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.

[0064] In some embodiments, in step (3), the centrifugation time is 5-30 minutes. In some embodiments, the centrifugation time is 5-10 minutes, 10-15 minutes, 15-20 minutes, 20-25 minutes or 25-30 minutes. In some embodiments, the centrifugation time is 10 minutes.

[0065] Step (5)

[0066] In some embodiments, in step (5), 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), 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%.

[0067] 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%.

[0068] In some embodiments, in step (5), the amount of water added is 5-50 times the amount of the precipitate collected in step (4). In some embodiments, in step (5), 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).

[0069] In some embodiments, in step (5), the precipitate collected in step (4) is heated and dissolved at a temperature of 40-80° C. In some embodiments, in step (5), 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), the precipitate collected in step (4) is heated and dissolved at a temperature of 60° C.

[0070] Step (6)

[0071] In some embodiments, in step (6), refrigeration is performed at a temperature of 0.1-10° C. In some embodiments, in step (6), 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), refrigeration is performed at a temperature of 2-8° C.

[0072] In some embodiments, in step (6), the refrigeration time is greater than 1 hour. In some embodiments, the refrigeration time is 1 hour to 3 months. In some embodiments, the refrigeration time is 1-2 hours, 2-3 hours, 3-4 hours, 4-5 hours, 5-6 hours, 6-7 hours, 7-8 hours, 8-9 hours, 9-10 hours, 10-11 hours, 11-12 hours, 12 hours to 1 day, 1-2 days, 2-5 days, 5-10 days, 10-15 days, 15-20 days, 20-25 days, 25 days to 1 month, 1-2 months, 2-3 months. In some embodiments, the refrigeration time is 2-12 hours.

[0073] Step (7)

[0074] In some embodiments, in step (7), the suspension obtained in step (6) is centrifuged and filtered. In some embodiments, in step (7), the suspension obtained in step (6) is first centrifuged and then filtered. In some embodiments, in step (7), the suspension obtained in step (6) is first centrifuged, then filtered, and then centrifuged. In some embodiments, in step (7), the suspension obtained in step (6) is centrifuged. In some embodiments, in step (7), 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.

[0075] Step (8)

[0076] In some embodiments, the drying in step (8) includes, but is not limited to, freeze drying, vacuum drying, atmospheric pressure drying, spray drying, boiling drying, or a combination thereof.

[0077] Rhodiola rosea polysaccharide extract

[0078] The second aspect of the present invention relates to a Rhodiola rosea polysaccharide extract, which is prepared by the method described in the first aspect of the present invention.

[0079] In some embodiments, the storage stability of the Rhodiola rosea polysaccharide extract is higher than that of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the storage stability of the Rhodiola rosea polysaccharide extract is higher than that of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0080] In some embodiments, the toxicity of the Rhodiola rosea polysaccharide extract to cells (e.g., HaCaT cells, RAW264.7 cells, etc.) is lower than that of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the toxicity of the Rhodiola rosea polysaccharide extract to cells (e.g., HaCaT cells, RAW264.7 cells, etc.) is lower than that of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0081] In some embodiments, the Rhodiola rosea polysaccharide extract can scavenge reactive oxygen free radicals produced by HaCaT cells induced by H2O2. In some embodiments, the Rhodiola rosea polysaccharide extract can scavenge reactive oxygen free radicals produced by cells induced by H2O2.

[0082] In some embodiments, the Rhodiola rosea polysaccharide extract has an antioxidant effect. In some embodiments, the antioxidant effect of the Rhodiola rosea polysaccharide extract is stronger than the antioxidant effect of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the antioxidant effect of the Rhodiola rosea polysaccharide extract is stronger than the antioxidant effect of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0083] In some embodiments, the Rhodiola rosea polysaccharide extract can inhibit LPS-induced IL-6 secretion in RAW264.7 cells. In some embodiments, the Rhodiola rosea polysaccharide extract can inhibit cells from secreting pro-inflammatory factors (eg, IL-6).

[0084] In some embodiments, the Rhodiola rosea polysaccharide extract has an anti-inflammatory effect. In some embodiments, the anti-inflammatory effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-inflammatory effect of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the anti-inflammatory effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-inflammatory effect of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0085] In some embodiments, the Rhodiola rosea polysaccharide extract can reduce the decrease in the expression level of type III collagen gene in HFF-1 cells induced by UVA. In some embodiments, the Rhodiola rosea polysaccharide extract can promote the expression of collagen genes (e.g., type III collagen genes) in cells. In some embodiments, the Rhodiola rosea polysaccharide extract can promote the expression of fibroblast collagen genes.

[0086] In some embodiments, the Rhodiola rosea polysaccharide extract has an anti-aging effect. In some embodiments, the anti-aging effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-aging effect of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the anti-aging effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-aging effect of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0087] In some embodiments, the Rhodiola rosea polysaccharide extract has an anti-photoaging effect. In some embodiments, the anti-aging effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-photoaging effect of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the anti-photoaging effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-photoaging effect of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0088] In some embodiments, the Rhodiola rosea polysaccharide extract can reduce the decrease in the expression level of CLDN1 gene in HaCaT cells induced by UVA. In some embodiments, the Rhodiola rosea polysaccharide extract can promote the expression of CLDN1 gene. In some embodiments, the Rhodiola rosea polysaccharide extract can promote the expression of epidermal cell barrier genes.

[0089] In some embodiments, the Rhodiola rosea polysaccharide extract has a skin repairing effect. In some embodiments, the skin repairing effect of the Rhodiola rosea polysaccharide extract is stronger than the skin repairing effect of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the skin repairing effect of the Rhodiola rosea polysaccharide extract is stronger than the skin repairing effect of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0090] In some embodiments, the Rhodiola rosea polysaccharide extract has an anti-photodamage effect. In some embodiments, the anti-photodamage effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-photodamage effect of the Rhodiola rosea polysaccharide extract of the prior art. In some embodiments, the anti-photodamage effect of the Rhodiola rosea polysaccharide extract is stronger than the anti-photodamage effect of the Rhodiola rosea polysaccharide extract obtained by the preparation method without the step of removing the precipitate after refrigeration after the water extraction and alcohol precipitation step.

[0091] Use of Rhodiola rosea polysaccharide extract in preparing cosmetics or medicines

[0092] The third aspect of the present invention relates to use of the Rhodiola rosea polysaccharide extract according to the second aspect of the present invention in the preparation of cosmetics or medicines.

[0093] In some embodiments, the cosmetic or medicine has antioxidant, anti-inflammatory, anti-acne, soothing, anti-aging, anti-photoaging, anti-photodamage and / or repair effects.

[0094] In some embodiments, the anti-acne effect is to combat inflammation caused by acne.

[0095] In some embodiments, the soothing effect is to combat skin inflammation caused by external stimuli.

[0096] Cosmetics or medicines

[0097] The fourth aspect of the present invention relates to a cosmetic or a medicine comprising the Rhodiola rosea polysaccharide extract according to the second aspect of the present invention.

[0098] In some embodiments, the composition accounts for 0.01%-20% by weight of the cosmetic or drug. In some embodiments, the composition 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%, 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%.

[0099] In some embodiments, the medicine or cosmetics are used for skin care. In some embodiments, the medicine and cosmetics are used for skin anti-oxidation, anti-inflammatory, anti-acne, soothing, anti-aging, anti-photoaging, anti-light damage and / or repair. In some embodiments, the medicine and cosmetics contain the Rhodiola rosea polysaccharide extract according to the second 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.

[0100] 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, and the like. DETAILED DESCRIPTION

[0101] The present invention can be implemented through the following embodiments, but the present invention is not limited thereto.

[0102] 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.

[0103] 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.

[0104] 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.

[0105] Experimental materials, equipment and statistical analysis methods

[0106] Experimental reagents: Rhodiola rosea was obtained from the Rhodiola rosea planting base of Shanghai Xiangyi Bencao Cosmetics Co., Ltd. in Suozhu Township, Tibet; D(+) anhydrous glucose standard (Shanghai Adamas Reagent Co., Ltd., 61055B); ZTC101 juice clarifier (Wuhan Zhengtiancheng Biotechnology Co., Ltd.); DMEM medium (Sigma-Aldrich (Shanghai) Trading Co., Ltd., D6429); FBS (Corning (Shanghai) Co., Ltd., 35-081-CV); double antibody (Sigma-Aldrich (Shanghai) Trading Co., Ltd., V900929); PBS (Sigma-Aldrich (Shanghai) Trading Co., Ltd., D8537); human fibroblasts and keratinocytes (purchased from the Cell Resource Center of Shanghai Institutes for Life Sciences, Chinese Academy of Sciences); RAW624.7 cells (Beijing Beina Chuanglian Biotechnology Research Institute, BNCC354753)

[0107] Experimental equipment: centrifuge (GQ105B vertical high-speed drum centrifuge, Shanghai Tianben Centrifuge Co., Ltd.); cell culture incubator (HERAcell 150i, Thermo); multifunctional microplate reader (Varioskan LUX, ThermoFisher); UV irradiator (UV-621111801, Genefi);

[0108] Statistical methods: The experimental results are the mean ± standard deviation of multiple parallel experimental data. Paired t-test was used to test whether the differences between groups were statistically significant. The statistical software was GraphPad 5.0.

[0109] Example

[0110] Preparation Example

[0111] Example 1

[0112] Take 100g of Rhodiola rosea medicinal material, crush it into 10 mesh coarse powder, add 0.6L of pure water, heat and reflux to extract for 1h, then filter, add 0.6L of pure water to the residue, heat and reflux to extract for 1h, then filter, combine the filtrate, concentrate and centrifuge at 10000r / min for 15min, add 3 times the volume of 95% ethanol to the supernatant, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, refrigerate at 2℃ for 2h, centrifuge at 10000r / min for 15min, take the supernatant, filter, concentrate, and freeze-dry to obtain Rhodiola rosea polysaccharide extract.

[0113] Example 2

[0114] Take 100g of Rhodiola rosea medicinal material, crush it into 10 mesh coarse powder, add 1L of pure water, heat reflux extraction for 1.5h, then filter, concentrate and centrifuge at 10000r / min for 15min, add 5 times volume of 95% ethanol to the supernatant, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, refrigerate at 5℃ for 6h, centrifuge at 10000r / min for 15min, take the supernatant, filter, concentrate, and freeze-dry to obtain Rhodiola rosea polysaccharide extract.

[0115] Example 3

[0116] Take 100g of Rhodiola rosea medicinal material, crush it into 10 mesh coarse powder, add 1.5L of pure water, heat reflux extraction for 2h, then filter, concentrate and centrifuge at 10000r / min for 15min, add 8 times volume of 95% ethanol to the supernatant, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, refrigerate at 8℃ for 12h, centrifuge at 10000r / min for 15min, take the supernatant, filter, concentrate, and freeze-dry to obtain Rhodiola rosea polysaccharide extract.

[0117] Comparative Example 1

[0118] Take 100g of Rhodiola rosea medicinal material, crush it into 10 mesh coarse powder, add 1L of pure water, heat reflux extraction for 1.5h, then filter, concentrate and centrifuge at 10000r / min for 15min, add 5 times volume of 95% ethanol to the supernatant, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, centrifuge at 10000r / min for 15min, take the supernatant, filter, concentrate, and freeze-dry to obtain Rhodiola rosea polysaccharide extract.

[0119] Comparative Example 2

[0120] Take 100g of Rhodiola rosea medicinal material, crush it into 10 mesh coarse powder, add 1L of pure water, heat reflux extraction for 1.5h and then filter, concentrate and centrifuge at 10000r / min for 15min, add 5 times the volume of 95% ethanol to the supernatant, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, centrifuge at 10000r / min for 15min, take the supernatant and filter, add 5 times the volume of 95% ethanol again, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, centrifuge at 10000r / min for 15min, take the supernatant and filter, concentrate, and freeze-dry to obtain the Rhodiola rosea polysaccharide extract.

[0121] Comparative Example 3

[0122] Take 100g of Rhodiola rosea medicinal material, crush it into 10 mesh coarse powder, add 1L of pure water, heat reflux extraction for 1.5h, then filter, concentrate and centrifuge at 10000r / min for 15min, add 5 times volume of 95% ethanol to the supernatant, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, add 0.1% ZTC101 flocculant, let stand and flocculate for 5min, the flocculation temperature is 25℃, centrifuge at 10000r / min for 15min, take the supernatant, filter, concentrate, and freeze-dry to obtain Rhodiola rosea polysaccharide extract.

[0123] Comparative Example 4

[0124] Take 100g of Rhodiola rosea medicinal material, crush it into 10 mesh coarse powder, add 1L of pure water, heat reflux extraction for 1.5h, then filter, concentrate and centrifuge at 10000r / min for 15min, add 5 times the volume of 95% ethanol to the supernatant, mix well and let stand for 2h for alcohol precipitation, centrifuge at 10000r / min for 10min, collect the precipitate, add pure water, heat at 60℃ to fully dissolve, and prepare a 10g / mL aqueous solution, mix the crude Rhodiola rosea polysaccharide extract and SP700 macroporous adsorption resin in a mass ratio of 0.1:1, place on a reciprocating oscillator and shake for 4 hours, rinse the macroporous resin with water 4 times the weight of the SP700 macroporous adsorption resin, filter and concentrate, freeze-dry after filtration to obtain a Rhodiola rosea polysaccharide extract.

[0125] Effect Example

[0126] Effect Example 1. Determination of polysaccharide content

[0127] Preparation of standard curve: accurately weigh 10mg D(+) anhydrous glucose standard into a 100mL volumetric flask, dissolve and dilute to volume with distilled water. Accurately pipette 0mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, and 1.0mL of solution into 6 groups of stoppered test tubes, 3 parallel in each group, add distilled water to 2.0mL, and mix. Accurately add 1.0mL of 5% phenol solution respectively, mix, quickly and accurately add 5.0mL of concentrated sulfuric acid, mix, place in a 40℃ water bath for 30min, take out, place in an ice water bath for 5min, take out, measure the absorbance at a wavelength of 490nm by UV-visible spectrophotometry, and draw a standard curve with absorbance as the ordinate and concentration as the abscissa.

[0128] Sample pretreatment: Accurately weigh 10 mg of the extract powder into a 100 mL volumetric flask, fully dissolve it in distilled water and dilute to volume. Determine it according to the above phenol-sulfuric acid method, and calculate the polysaccharide content in the sample solution according to the standard curve.

[0129] The polysaccharide content in the Rhodiola rosea extracts prepared in Examples 1-3 of the present invention and Comparative Examples 1-4 was quantitatively determined according to the above method, and the results are shown in Table 1 below.

[0130] Table 1

[0131] Sample No. Polysaccharide content (%) Example 1 63.28 Example 2 65.49 Example 3 66.34 Comparative Example 1 62.42 Comparative Example 2 57.86 Comparative Example 3 56.49 Comparative Example 4 67.82

[0132] It can be seen from the results in Table 1 that Examples 1-3 of the present invention and Comparative Examples 1-4 all successfully obtained Rhodiola rosea polysaccharide extracts with polysaccharides as the main component.

[0133] Effect Example 2. Yield and Stability

[0134] The prepared Rhodiola rosea polysaccharide extracts of Examples 1-3 and Comparative Examples 1-4 were weighed and prepared into 5% aqueous solutions, and the color and state of the aqueous solutions containing 5% Rhodiola rosea polysaccharide extracts were observed. The weight, yield, and color and state of the 5% aqueous solutions of the Rhodiola rosea polysaccharide extracts prepared by different processes are recorded in Table 2 below.

[0135] Table 2

[0136]

[0137]

[0138] As shown in Table 2, the yield of the Rhodiola rosea polysaccharide extract obtained by the extraction method of the present invention is as high as 8% or more. Compared with the traditional water extraction and alcohol precipitation polysaccharide extraction method, the clarity of the extract in the aqueous solution is greatly improved and the precipitation of the solution after being placed at room temperature for 48 hours is reduced. In contrast to Examples 2, 3 and 4, although the color is improved to a certain extent, multiple alcohol precipitation, flocculation and macroporous resin adsorption purification methods not only fail to improve the clarity of the Rhodiola rosea polysaccharide extract, but also greatly reduce the yield of the extract. Therefore, compared with the method in Comparative Examples 1-4, the extraction method of the present invention significantly improves the yield of the extract while improving the clarity and transparency of the Rhodiola rosea polysaccharide extract and reducing its precipitation.

[0139] Effect Example 3. Evaluation of Antioxidant Capacity

[0140] 3.1 Experimental methods

[0141] The antioxidant capacity of the sample is evaluated by evaluating the sample's ability to scavenge intracellular reactive oxygen free radicals (ROS). The specific operation scheme is as follows:

[0142] Add 100 μL of 1.5 × 10 5 HaCaT cells of 100 μg / mL were incubated in an incubator at 37°C and 5% CO2 overnight. The culture medium was removed, and the samples of Examples 1-3 and Comparative Examples 1-4 were added to the drug administration group at a concentration of 200 μg / mL, 100 μg / mL, 50 μg / mL, and 20 μg / mL in DMEM containing 10% FBS, and the blank group and model group were added to DMEM containing 10% FBS. The positive control group was added to epigallocatechin gallate (EGCG) at a concentration of 20 μg / mL in DMEM containing 10% FBS. After being placed in the incubator for 16 hours, 100 μL of DMEM containing 20 μM DCFH-DA solution and 5 μg / mL Hoechst was added to each well and placed in the incubator for 20 minutes. PBS was rinsed once, and 100 μL of H2O2 (100 μM) prepared in PBS was added to each well except the blank group, and placed in the incubator for 20 minutes. Intracellular ROS can oxidize non-fluorescent DCFH to generate fluorescent DCF, and the intensity of fluorescence is proportional to the level of ROS. Hoechst is used to stain cell nuclei and detect living cells to determine the safe concentration of the sample.

[0143] 3.2 Experimental Results

[0144] The experimental results of Hoechst staining cell nuclei to detect living cells are as follows Figure 1As shown, Examples 1-3 did not show obvious cytotoxic effects at concentrations of 20 μg / mL to 200 μg / mL, while Comparative Examples 1-4 showed obvious cytotoxic effects at 100 μg / mL and 200 μg / mL. Therefore, compared with Comparative Examples 1-4, Examples 1-3 significantly reduced the cytotoxicity and irritation to human keratinocytes, thereby improving the safety of the Rhodiola rosea polysaccharide extract. At safe concentrations of 50 μg / mL and 20 μg / mL, Examples 1-3 and Comparative Examples 1-4 were compared to see the inhibitory effects on the production of ROS by HaCaT cells induced by H2O2. The results are as follows: Figure 2 As shown, Examples 1-3 showed strong inhibitory effects on the production of ROS by HaCaT cells induced by H2O2 at concentrations of 50 μg / mL and 20 μg / mL. Except for Comparative Examples 1 and Comparative Examples 4, which showed weak inhibitory effects on the production of ROS by HaCaT cells induced by H2O2 at a concentration of 50 μg / mL, Comparative Examples 2 and 3 at concentrations of 50 μg / mL and 20 μg / mL, and Comparative Examples 1 and 4 at a concentration of 20 μg / mL did not show inhibitory effects on ROS. The above results show that, compared with Comparative Examples 1-4, Examples 1-3 significantly improved the inhibitory effect on the production of ROS by HaCaT cells induced by H2O2, while significantly reducing the irritation and toxicity to HaCaT cells.

[0145] Effect Example 4. Evaluation of anti-inflammatory ability

[0146] 4.1 Experimental methods

[0147] The anti-inflammatory ability of the samples was evaluated by evaluating the inhibitory effect of the samples on the IL-6 factor secreted in RAW264.7 cells. The specific operation scheme is as follows:

[0148] Add 100 μL of 2.5×10 5 RAW264.7 cells of 100 μg / mL, 50 μg / mL, 20 μg / mL samples of Examples 1-3 and Comparative Examples 1-4 prepared with DMEM containing 10% FBS and lipopolysaccharide (LPS) at a final concentration of 1 μg / mL were added to the administration group, DMEM containing 10% FBS was added to the blank group, and LPS at a final concentration of 1 μg / mL was added to the model group, and culture was continued for 24 hours. After the culture was completed, the cell supernatant was collected, and the pro-inflammatory factor IL-6 was detected using the Mouse IL-6 ELISA kit. 100 μL of 10% CCK-8 solution was added to the cells in the well plate, and the cells were placed in a cell culture incubator for 30 minutes. The light absorption value was detected at OD 450nm of the microplate reader, and the effect of the sample to be tested on the cell viability was calculated.

[0149] 4.2 Experimental Results

[0150] According to the RAW264.7 cell viability test results, Figure 3 As shown, Examples 1-3 did not show obvious cytotoxic effects at concentrations of 100 μg / mL to 20 μg / mL, while Comparative Examples 1-4 showed a certain inhibitory effect on cell viability at a concentration of 100 μg / mL. Therefore, safe concentrations of 50 μg / mL and 20 μg / mL were selected to study the inhibitory effect on LPS-induced secretion of inflammatory factor IL-6 by RAW264.7 cells. The results are shown in FIG. Figure 4 As shown. Figure 4 The results show that, compared with comparative examples 1-4, examples 1-3 have a stronger inhibitory effect on LPS-induced IL-6 secretion by RAW264.7 cells at concentrations of 50 μg / mL and 20 μg / mL, and have a certain dose-effect relationship, indicating that the stimulating effect of the Rhodiola rosea polysaccharide extract of examples 1-3 on RAW264.7 cells is significantly lower than that of the Rhodiola rosea polysaccharide extract obtained in comparative examples 1-4, and has a stronger anti-inflammatory effect.

[0151] Effect Example 5. Evaluation of anti-aging ability

[0152] 5.1 Experimental Methods

[0153] The anti-aging ability of the samples was evaluated based on the expression level of type III collagen gene in the photoaging model of HFF-1 cells induced by UVA. The specific operation plan is as follows:

[0154] HFF-1 cells were cultured at 1×10 5 The cells were seeded into 24-well plates at a density of 100 cells / mL. After the cells adhered to the plate the next day, 8 J / cm 2 UVA irradiation treatment, the drug-treated group was prepared with DMEM complete medium at a concentration of 50 μg / mL of Examples 1-3 and Comparative Examples 1-4, the blank control group was not subjected to UVA irradiation treatment, and the model control group was only irradiated without drug administration. After continuous culture for 16 hours, the cells were lysed and the total cell RNA was collected. RNA was extracted and purified using the SteadyPure Universal RNA Extraction Kit, and RT-qPCR reaction was performed after reverse transcription to obtain cDNA to detect type III collagen gene expression.

[0155] 5.2 Experimental Results

[0156] The effects of Examples 1-3 and Comparative Examples 1-4 on the UVA-induced HFF-1 cell photoaging model were as follows: Figure 5 As shown by Figure 5The results show that UVA irradiation can significantly inhibit the expression of type III collagen gene in HFF-1 fibroblasts. Compared with the model group, the Rhodiola rosea polysaccharide extract of Examples 1-3 at a concentration of 50 μg / mL can significantly improve the decrease in type III collagen gene expression caused by UVA, indicating that the polysaccharide extract of Examples 1-3 has a significantly stronger promoting effect on type III collagen gene expression than that of Comparative Examples 1-4, indicating that the Rhodiola rosea polysaccharide extract obtained by the process of Examples 1-3 has significant advantages in anti-photoaging and anti-aging.

[0157] Effect Example 6. Evaluation of Anti-photodamage and Skin Repair Ability

[0158] 6.1 Experimental methods

[0159] CLDN1 (Claudin-1) is a transmembrane protein that makes up the tight junctions of the epidermis. Tight junctions are composed of adjacent cell membranes and play an important role in maintaining the permeability barrier and cell polarity. They are considered to be the main control factor of the skin barrier. Therefore, CLDN1 is related to regulating the permeability of the skin barrier structure and helps maintain normal skin barrier function.

[0160] The ability of the samples to resist light damage and repair skin was evaluated based on the CLDN1 gene expression level of HaCaT cells induced by UVA. The specific operation plan is as follows:

[0161] HaCaT cells were cultured at 2×10 5 The cells were seeded into 24-well plates at a density of 100 cells / mL. After the cells adhered to the wall the next day, 0.2 J / cm 2 UVA irradiation treatment, the drug administration group was prepared with DMEM complete medium at a concentration of 50 μg / mL Example 1-3 and Comparative Example 1-4, the blank control group was not subjected to UVA irradiation treatment, and the model control group was only irradiated without drug administration. After continuous culture for 24 hours, the cells were lysed and the total cell RNA was collected. RNA was extracted and purified using RNA Easy Fast Tissue / Cell Kit, and RT-qPCR reaction was performed after reverse transcription to detect CLDN1 gene expression.

[0162] 6.2 Experimental Results

[0163] The effects of Examples 1-3 and Comparative Examples 1-4 on the UVA-induced HaCaT cell model are shown in the following table. Figure 6 As shown by Figure 6The results show that UVA irradiation can significantly inhibit the expression of CLDN1 gene in HaCaT cells. Compared with the model group, the Rhodiola rosea polysaccharide extract of Examples 1-3 at a concentration of 50 μg / mL can significantly improve the decrease in the CLDN1 gene level in HaCaT cells caused by UVA, and its promoting effect on CLDN1 gene expression is significantly better than the Rhodiola rosea polysaccharide extract obtained in Comparative Examples 1-4, indicating that the Rhodiola rosea polysaccharide extract obtained by the process of Examples 1-3 has significant advantages in the direction of anti-photodamage and repair, and the present invention is the first to discover the advantages of Rhodiola rosea polysaccharide in the direction of skin repair.

Claims

1. A method for preparing a Rhodiola rosea polysaccharide extract, the method comprising the following steps: (1) adding the Rhodiola rosea raw material into 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; (3) adding alcohol to the filtrate obtained in the optionally concentrated step (2), mixing evenly and letting stand for a period of time to obtain an alcohol precipitate; (4) centrifuging the alcohol precipitate obtained in step (3) and collecting the precipitate; (5) adding the precipitate collected in step (4) into water and heating until fully dissolved; (6) refrigerating the solution obtained in 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.

2. The method according to claim 1, wherein the method further comprises the following steps: (8) Drying the optionally concentrated supernatant or filtrate obtained in step (7).

3. The method according to claim 1 or 2, wherein the Rhodiola rosea is a plant of the genus Rhodiola, preferably selected from one or more of Rhodiola crenulata (Hook.f.et Thoms.) H.Ohba, Rhodiola rosea L., Rhodiola sacra (Prain ex Raym.-Hamet) SHFu, Rhodiola fastigiata (Hook.f.&Thomson) SHFu, and Rhodiola dumulosa (Franch.) SHFu, most preferably Rhodiola crenulata (Hook.f.et Thoms.) H.Ohba.

4. The method according to any one of claims 1 to 3, wherein in step (1), the weight ratio of Rhodiola rosea raw material to water is 1:5 to 1:30, the number of extractions is 1-3 times, and each extraction is 0.5-5h, preferably 0.5-3h.

5. The method according to any one of claims 1 to 4, wherein in step (3), the alcohol used is ethanol or methanol, preferably ethanol; the concentration and volume of the added alcohol are such that the concentration of alcohol in the alcohol precipitation solution obtained after the addition of alcohol is 40% to 90%; the standing time is 0.5-36h, preferably 0.5-12h, most preferably 2-8h.

6. The method according to any one of claims 1 to 5, wherein in step (5), the precipitate collected in step (4) is dissolved by heating at a temperature of 40-80°C.

7. The method according to any one of claims 1 to 6, wherein in step (6), refrigeration is carried out at a temperature of 0.1-10°C, preferably 2-8°C; the refrigeration time is greater than 1 hour, preferably 2-12 hours.

8. A Rhodiola rosea polysaccharide extract, which is prepared by the method according to any one of claims 1 to 7.

9. Use of the Rhodiola rosea polysaccharide extract according to claim 8 in the preparation of cosmetics or medicines.

10. The use according to claim 9, wherein the cosmetic or medicine has antioxidant, anti-inflammatory, anti-acne, soothing, anti-aging, anti-photoaging, anti-photodamage and / or repairing effects.

11. A cosmetic or a medicine comprising the Rhodiola rosea polysaccharide extract according to claim 9.

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