Method for extracting roselle, roselle extract, food composition, pharmaceutical composition or skincare cosmetic containing roselle extract, and use of roselle extract and food composition, pharmaceutical composition or skincare cosmetic containing roselle extract

Abstract

The invention discloses a roselle extraction method, a roselle extract, a food composition, a pharmaceutical composition or a skin care cosmetic containing the roselle extract and application of the roselle extract. The method for extracting the roselle comprises the following steps: providing dried roselle as an extraction raw material; carrying out fine crushing on the extracted raw materials; mixing the finely crushed extraction raw material with an extraction solvent, and carrying out primary extraction; performing ultrasonic extraction on the primary extract of the roselle; and filtering the roselle extract.

Description

Technical Field

[0001] The invention relates to an extraction method, in particular to a method for extracting black crystal roselle, an extract obtained by using the extraction method and application thereof. Background Art

[0002] Hibiscus sabdariffa, also known as Roselle, Roselle Flower, and Mountain Solanum, is a one- or two-year-old herb distributed in tropical and subtropical regions. The most commonly eaten part is the calyx, which is often made into candied fruit, jam, etc., and can also be used to make tea.

[0003] It is known that Roselle contains organic acids, carbohydrates, carotene, anthocyanins and other ingredients, and studies have shown that Roselle has anti-cancer, anti-inflammatory, cholesterol and triglyceride lowering, blood pressure regulating, and liver protection effects.

[0004] Therefore, how to improve the extraction method to enhance the active substances of Roselle or its related varieties and thus enhance their efficacy, or discover new uses for them, has become one of the important issues that this business wants to solve. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a method for extracting black crystal roselle, an extract obtained by the extraction method and uses thereof in view of the deficiencies in the prior art.

[0006] In order to solve the above-mentioned technical problems, one of the technical solutions adopted by the present invention is to provide a method for extracting black crystal Roselle, which includes step S1: providing dried black crystal Roselle as an extraction raw material; step S2: finely grinding the extraction raw material; step S3: mixing the finely ground extraction raw material with an extraction solvent for primary extraction; step S4: ultrasonically extracting the primary extract of black crystal Roselle; and step S5: filtering the black crystal Roselle extract.

[0007] In order to solve the above technical problems, another technical solution adopted by the present invention is to provide a black crystal roselle extract prepared by the above-mentioned black crystal roselle extraction method.

[0008] In order to solve the above technical problems, another technical solution adopted by the present invention is to provide a food composition comprising the above-mentioned Roselle melanogaster extract.

[0009] In order to solve the above technical problems, another technical solution adopted by the present invention is to provide a pharmaceutical composition comprising the above-mentioned Roselle melanogaster extract.

[0010] In order to solve the above technical problems, another technical solution adopted by the present invention is to provide a skin care cosmetic containing the above-mentioned Roselle melanin extract.

[0011] In order to solve the above technical problems, another technical solution adopted by the present invention is to provide a use of the black crystal roselle extract obtained by the above-mentioned black crystal roselle extraction method for preparing a pharmaceutical composition for skin care.

[0012] One of the beneficial effects of the present invention is that the black crystal roselle extraction method, the extract using the extraction method and the use thereof provided by the present invention can improve the active substance components contained in the black crystal roselle extract through the technical scheme of "step S1: providing dried black crystal roselle as an extraction raw material; step S2: finely grinding the extraction raw material; step S3: mixing the finely ground extraction raw material and the extraction solvent for primary extraction; step S4: ultrasonically extracting the black crystal roselle primary extract; and step S5: filtering the black crystal roselle extract".

[0013] Furthermore, the black crystal roselle extract prepared by the above technical solution of the present invention exhibits beneficial effects such as inhibiting tyrosinase activity, reducing melanin content, increasing elastin content, and protecting cells from damage caused by UVA. In addition, the black crystal roselle extract prepared by the black crystal roselle extraction method of the present invention has beneficial effects on skin care.

[0014] To further understand the features and technical contents of the present invention, please refer to the following detailed description and drawings of the present invention. However, the drawings provided are only for reference and description and are not intended to limit the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The present invention is a flowchart of a method for extracting Roselle rubrum according to an embodiment of the present invention. Figure 2 The present invention is a flowchart of a method for extracting Roselle rubrum according to another embodiment of the present invention.

[0016] Figure 3 This is the HPLC spectrum of the black crystal Roselle extract obtained by the black crystal Roselle extraction method according to the first embodiment of the present invention.

[0017] Figure 4 This is the HPLC spectrum of the extract of the commercial variety Taidong No. 3 of Roselle of the first embodiment of the present invention.

[0018] Figure 5 This is an analysis chart of the effect of the black crystal roselle extract of the second embodiment of the present invention on the tyrosinase activity in melanoma cells. When there is no duplication in the histograms of each group, it means there is a significant difference (P≤0.05).

[0019] Figure 6This is an analysis chart of the effect of the black crystal roselle extract of the second embodiment of the present invention on the melanin content in melanoma cells. When there is no duplication in the English words on the histograms of each group, it means there is a significant difference (P≤0.05).

[0020] Figure 7 This is an analysis chart of the effect of the black crystal roselle extract of the third embodiment of the present invention on increasing the elastin content in fibroblasts. When there is no duplication in the histograms of each group, it means there is a significant difference (P≤0.05).

[0021] Figure 8 This is a cell survival rate analysis chart of the black crystal roselle extract in the fourth embodiment of the present invention for enhancing the UV resistance of fibroblasts. When there is no duplication in the histograms of each group, it means there is a significant difference (P≤0.05). DETAILED DESCRIPTION

[0022] The following is an explanation of the implementation methods of the "Method for extracting black crystal roselle, extracts using the extraction method and uses thereof" disclosed in the present invention through specific embodiments. Those skilled in the art can understand the advantages and effects of the present invention from the contents disclosed in this specification. The present invention can be implemented or applied through other different specific embodiments, and the details in this specification can also be modified and changed in various ways based on different viewpoints and applications without departing from the concept of the present invention. In addition, the drawings of the present invention are only simple schematic illustrations and are not depicted in actual size. It is stated in advance. The following embodiments will further explain the relevant technical contents of the present invention in detail, but the disclosed contents are not intended to limit the scope of protection of the present invention. In addition, the term "or" used in this article may include any one or more combinations of the associated listed items depending on the actual situation.

[0023] See also Figure 1 and Figure 2 As shown, the present invention provides a method for extracting black crystal Roselle, which at least includes step S1: providing dried black crystal Roselle as an extraction raw material; step S2: finely grinding the dried black crystal Roselle extraction raw material; step S3: mixing black crystal Roselle powder with an extraction solvent for primary extraction; step S4: ultrasonically extracting the primary black crystal Roselle extract; and step S5: filtering and extracting a crude black crystal Roselle extract. It should be noted that the crude black crystal Roselle extract of the present invention can be directly used as a black crystal Roselle extract without performing other extraction treatment procedures, but the present invention is not limited thereto.

[0024] In one embodiment, the method for extracting Roselle nigra of the present invention further comprises step S6: filtering the crude Roselle nigra extract to obtain the Roselle nigra extract.

[0025] In one embodiment, the black crystal roselle extract of the present invention can be taken in an edible form, and the black crystal roselle extract can be made into a food composition of the black crystal roselle extract in the form of liquid, solid, granular, powder, jelly or gel. The food composition of the black crystal roselle extract can be combined with an excipient or a flavoring agent to enhance its flavor or convenience of consumption.

[0026] The excipients used in the food composition of the black crystal roselle extract can be, for example, corn starch, potato starch, dextrin, lactose, glucose, fructose, sorbitol, xytol, mannitol, etc., which can be adjusted according to needs and practical applications, and the present invention is not limited thereto.

[0027] The flavoring agent used in the food composition of the black crystal roselle extract can be, for example, fruit juice extract, sucralose, mannose, xylitol, citric acid, malic acid, tartaric acid, etc., which can be adjusted according to needs and practical applications, and the present invention is not limited thereto.

[0028] In one embodiment, the black crystal roselle extract of the present invention has the beneficial effects of inhibiting tyrosinase activity, reducing melanin content, increasing elastin content, and protecting cells from damage caused by UVA. Therefore, the black crystal roselle extract of the present invention can be further formulated into a pharmaceutical composition for skin care. Therefore, the present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the black crystal roselle extract and one or more pharmaceutically acceptable carriers.

[0029] For the purpose of delivery and absorption, the therapeutically effective amount of the black crystal roselle extract of the present invention can be formulated into a suitable form of pharmaceutical composition for skin care together with a pharmaceutically acceptable carrier. According to the administration route, the black crystal roselle extract pharmaceutical composition of the present invention preferably contains 0.1% to 100% of the black crystal roselle extract by total weight.

[0030] In one embodiment, the pharmaceutically acceptable carrier can be an acceptable carrier, diluent or excipient that is compatible with other ingredients in the formulation and harmless to the individual to whom the pharmaceutical composition is administered. Any carrier, diluent or excipient commonly known or used in the art can be used in the present invention, depending on the requirements of the pharmaceutical formulation. According to the present invention, the pharmaceutical composition can be used for administration via any appropriate route, including but not limited to oral, rectal, nasal, topical, vaginal or parenteral routes. In a specific example of the present invention, the pharmaceutical composition is formulated for oral administration. Such preparations can be prepared by any method known in the pharmaceutical field.

[0031] In one embodiment, the pharmaceutical composition of the black crystal roselle extract of the present invention can be in the form of tablets, pills, powders, buccal tablets, packets, buccal lozenges, elixirs, suspensions, lotions, solutions, syrups, soft and hard gelatin capsules, suppositories, sterile injections, and packaged powders, but the present invention is not limited thereto.

[0032] In one embodiment, the Roselle melanogaster extract of the present invention can be further formulated for use in the manufacture of skin care cosmetics, wherein the skin care cosmetics have whitening, anti-allergy, and anti-aging effects.

[0033] In one embodiment, the black crystal roselle extract of the present invention can be further made into an external preparation suitable for topical application to the skin when formulated for use in the manufacture of skin care cosmetics, including but not limited to emulsifiers, gels, creams, creams, patches, ointments, powders, aerosols, sprays, lotions, serums, pastes, foams, drops, suspensions, ointments and bandages.

[0034] [First embodiment]

[0035] See also Figure 1 and Figure 2 As shown, the first embodiment of the present invention provides a method for extracting black crystal roselle, which at least comprises the following steps:

[0036] Step S1: Provide dried black crystal Roselle as an extraction raw material. In one embodiment, the black crystal Roselle extraction raw material is dried black crystal Roselle, preferably, the black crystal Roselle extraction raw material is dried black crystal Roselle with a moisture content of less than 10%. Black crystal Roselle can be dried by sun drying, hot air drying, low temperature cold air drying, freeze drying, vacuum drying and the like, which can be adjusted according to needs and practical applications, and the present invention is not limited. It is worth noting that before drying, the black crystal Roselle extraction raw material can be pre-treated to promote the drying effect or prevent the deterioration of the black crystal Roselle components during the drying process. For example, the black crystal Roselle extraction raw material can be blanched in boiling water to kill the cyanine, thereby inhibiting the enzyme activity contained in the black crystal Roselle. The pre-treatment can be adjusted according to needs and practical applications, and the present invention is not limited.

[0037] Step S2: Finely pulverize the dried black crystal Roselle extract material. In one embodiment, the dried black crystal Roselle extract material can be beaten into fine powder by a grinder, and homogenized by a 100-mesh screen to obtain black crystal Roselle powder. However, the present invention is not limited to this, and a homogenizer or other homogenization method can be selected according to conventional technology to finely pulverize the black crystal Roselle extract material. In addition, the mesh size of the screen can also be selected according to actual application, and the present invention is not limited thereto.

[0038] Step S3: Mix the black crystal Roselle powder with the extraction solvent for initial extraction. In one embodiment, the extraction solvent may be water and / or a water-soluble organic solvent. The water-soluble organic solvent may be a mixture of one or more of methanol, ethanol, n-propanol, isopropanol, butanol, ethylene glycol, propylene glycol, and glycerol, but the present invention is not limited thereto. In a preferred embodiment, the extraction solvent of black crystal Roselle is water, and the mixing ratio of black crystal Roselle and water may be 1:1 to 1:30, preferably 1:10 to 1:20, and more preferably 1:15, but the present invention is not limited thereto. Further, after the black crystal Roselle powder is mixed with the extraction solvent, the black crystal Roselle powder may be uniformly dissolved in the extraction solvent by standing, stirring, shaking, and the like. The mixing method may be adjusted according to demand and practical application, and the present invention is not limited thereto. In addition, after the black crystal Roselle powder is mixed with the extraction solvent, it may be heated for initial extraction to obtain the black crystal Roselle primary extract. For example, the temperature can be heated to 100°C, 95°C, 90°C, 85°C, 80°C, 75°C, 70°C, 65°C, 60°C, 55°C, 50°C, 45°C, 40°C, 35°C, etc. for initial extraction. In a preferred embodiment, the initial extraction temperature is 60°C. In addition, the initial extraction time can be adjusted according to demand and practical application, and the present invention is not limited thereto. For example, the initial extraction time can be 15 minutes, 30 minutes, 45 minutes, 60 minutes, 75 minutes, 90 minutes, 120 minutes, 180 minutes, 240 minutes, etc. In a preferred embodiment, the initial extraction time is 15 minutes to 60 minutes, and more preferably, the initial extraction time is 30 minutes. It is worth noting that after performing the above-mentioned initial extraction step, the black crystal roselle initial extract can be directly used as a black crystal roselle extract, but the present invention is not limited thereto.

[0039] Step S4: ultrasonic extraction of the first extract of Roselle. In detail, after the above step S3, the first extract of Roselle can be squeezed and filtered, and the remaining solid residue that cannot be filtered (hereinafter referred to as Roselle residue) is ultrasonically extracted. In one embodiment, the extraction solvent can be water and / or a water-soluble organic solvent. The water-soluble organic solvent can be a mixture of one or more of methanol, ethanol, n-propanol, isopropanol, butanol, ethylene glycol, propylene glycol, and glycerol, but the present invention is not limited thereto. In a preferred embodiment, the extraction solvent of Roselle residue is water, and the mixing ratio of Roselle residue to water can be 1:1 to 1:30, preferably 1:10 to 1:20, and more preferably 1:2.5 to 1:5, but the present invention is not limited thereto. In addition, after the Roselle residue is mixed with the extraction solvent, it can be heated and ultrasonically extracted to obtain the Roselle ultrasonic extract. For example, the ultrasonic extraction can be performed by heating to 100°C, 95°C, 90°C, 85°C, 80°C, 75°C, 70°C, 65°C, 60°C, 55°C, 50°C, 45°C, 40°C, 35°C, etc. In a preferred embodiment, the ultrasonic extraction temperature is 60°C. In one embodiment, the ultrasonic extraction can be performed on the black crystal Roselle residue mixed with the extraction solvent under ultrasonic conditions of 200W power and 40KHz frequency. In addition, the ultrasonic extraction time can be adjusted according to demand and practical application, and the present invention is not limited. For example, the ultrasonic extraction can be 15 minutes, 30 minutes, 45 minutes, 60 minutes, 75 minutes, 90 minutes, 120 minutes, 180 minutes, 240 minutes, etc. In a preferred embodiment, the ultrasonic extraction time is 15 minutes to 60 minutes, and more preferably, the ultrasonic extraction time is 30 minutes. It is worth noting that after the ultrasonic extraction step, the Roselle melanin ultrasonic extract can be directly used as the Roselle melanin extract, but the present invention is not limited thereto.

[0040] Step S5: Filtering the black crystal Roselle extract. As described above, the black crystal Roselle primary extract obtained after step S3 can be used directly as the black crystal Roselle extract, but the black crystal Roselle primary extract can also be subjected to the filtration step of step S5. In addition, if step S4 is implemented successively after step S3, the black crystal Roselle ultrasonic extract can be mixed with the black crystal Roselle primary extract and then subjected to the filtration step of step S5, but the black crystal Roselle ultrasonic extract can also be separated from the black crystal Roselle primary extract and then mixed together after the filtration step of step S5, which is not limited by the present invention. In one embodiment, the filtration step can be performed by squeezing and filtering the black crystal Roselle primary extract and / or the black crystal Roselle ultrasonic extract to obtain the black crystal Roselle extract. For example, squeezing and filtration can be implemented using a filter press, but the present invention is not limited thereto. In addition, the filtration step can be filtered by a filter cloth or a filter screen. In a preferred embodiment, the filter cloth can be a 20 mesh, 40 mesh, 60 mesh, 80 mesh, 100 mesh, 200 mesh, 300 mesh, 400 mesh filter cloth, more preferably, the embodiment of the present invention uses a 100 mesh filter cloth, but the present invention is not limited. It is worth noting that the residual solids can also be separated by centrifugation to obtain the black crystal roselle extract, which is not limited by the present invention.

[0041] It is worth noting that the black crystal roselle extract can be further filtered through a filter membrane, heat-treated, added with protease inhibitors, or added with preservatives to maintain the integrity and activity of the components of the black crystal roselle extract, but the present invention is not limited thereto.

[0042] Step S6: Drying the black crystal Roselle extract. It is worth noting that the black crystal Roselle extract obtained after step S5 can also be used directly without step S6. In one embodiment, the black crystal Roselle extract can be further dried to form a powdered black crystal Roselle extract, and the drying method can be, for example, but not limited to, spray drying, vacuum freeze drying, microwave vacuum drying or low-temperature cold air drying.

[0043] In addition, it should be noted that the black crystal roselle extract of the present invention can also be extracted by conventional extraction methods, as long as the extraction efficiency and the efficacy of the black crystal roselle extract can be achieved. For example, the extraction method can be by adjusting the pH value of the extraction environment and / or adding an extraction auxiliary agent, but the present invention is not limited thereto.

[0044] In one embodiment, the black crystal roselle extract was further subjected to component analysis. In this embodiment, the high performance liquid chromatography (HPLC) instrument used was an Agilent 1100 series system, which included a Degasser G1379A, a QuatPump G1311A, an Agilent Manual FL-Injection Valve injector, and a VWD G1314A variable wavelength detector. The analysis conditions are shown in Table 1 below.

[0045] Table 1. HPLC analysis conditions

[0046]

[0047] In addition, in this embodiment, cyanidin and delphinidin were used as standard substances and diluted to 0.025, 0.05, 0.1, 0.2, and 0.4 mg / L for use as standard solutions for HPLC analysis. The analysis results were used to prepare a standard curve to further estimate the content of cyanidin and delphinidin in the black crystal roselle extract.

[0048] This example also provides a comparative analysis of the current commercial variety of Roselle, Taidong No. 3 (hereinafter referred to as the control group) and the black crystal Roselle extract obtained by the extraction method of the present invention (hereinafter referred to as the experimental group). The extraction method and HPLC analysis method of the control group are the same as those of the experimental group, and will not be repeated.

[0049] like Figure 3 and Figure 4 As shown, after HPLC analysis of the extracts of the control group and the experimental group, the HPLC spectra of the components contained therein are substantially the same. In addition, through calculation of the standard curve, the Roselle extract of the control group contains 0.26 μg / mL of cyanidin and 0.9 μg / mL of delphinidin, while the black crystal Roselle extract of the experimental group contains 2 μg / mL of cyanidin and 5 μg / mL of delphinidin, indicating that the content of cyanidin and delphinidin in the black crystal Roselle extract of the present invention is much higher than that of the current commercial varieties of Roselle.

[0050] However, the above example is only one feasible embodiment and is not intended to limit the present invention.

[0051] [Second embodiment]

[0052] In this example, the ability of the Roselle serrata extract to inhibit tyrosinase activity was evaluated by tyrosinase activity assay, and the method is as follows.

[0053] It should be noted that in order to understand whether the concentration of Roselle melanin extract used in this example is toxic to melanoma cells, a cell viability analysis was first performed, and it was confirmed that the concentrations of Roselle melanin extract used (200 μg / mL, 400 μg / mL, and 800 μg / mL) were not cytotoxic (not shown).

[0054] Melanoma cells B16-F10 (1×10 5 ) were planted in a 24-well culture dish and cultured at 37°C for 24 hours. The cells were treated with different concentrations of black crystal roselle extract (the black crystal roselle extract recorded below is the black crystal roselle extract obtained after the above extraction step S6) for 48 hours, then washed with phosphate-buffered saline (PBS) and centrifuged at 10000g for 20 minutes at 4°C to collect the cell precipitate. The cell precipitate was resuspended in PBS containing 1% Triton X-100 and 100μM PMSF, and centrifuged at 17500g for 10 minutes at 4°C to remove cell debris, 90μl of cell lysate was added to 10μl of L-Dopa (2mg / ml), and then cultured at 37°C for 20 minutes to 30 minutes. The absorbance at 490nm was measured by a spectrophotometer. In addition, this embodiment also uses kojic acid as a positive control group, and its cell treatment method and tyrosinase activity analysis method are the same as above, which will not be repeated.

[0055] like Figure 5 As shown in Figure 2, when kojic acid was used to treat melanoma cells, the tyrosinase activity of melanoma cells was significantly inhibited. On the other hand, when 200 μg / mL, 400 μg / mL, and 800 μg / mL of black crystal roselle extract were used to treat melanoma cells, the tyrosinase activity of melanoma cells was also significantly inhibited. It is worth noting that, Figure 5 As shown, 400 μg / mL of the black crystal roselle extract showed a more significant inhibitory effect than kojic acid.

[0056] Furthermore, in this example, the ability of the Roselle spatholobi extract to inhibit melanin was evaluated by melanin content analysis, and the method is as follows.

[0057] Melanoma cells B16-F10 (1×10 5) were seeded in a 24-well culture dish and co-cultured at 37°C for 24 hours with the addition of 500 nM α-melanocyte stimulating hormone (α-MSH). The cells were treated with different concentrations of Roselle extract for 48 hours, washed with PBS and treated with trypsin. The cell pellet was collected after centrifugation at 10,000 g for 20 minutes at room temperature and lysed in 1N NaOH for 60 minutes. The melanin content was analyzed by comparing the absorbance of the test sample at 405 nm with the standard curve obtained using synthetic melanin.

[0058] like Figure 6 As shown, when melanoma cells were co-cultured with α-melanocyte stimulating hormone alone, the melanin content of melanoma cells increased significantly by about 2 times compared with the untreated control group, while the melanin content of melanoma cells was significantly reduced when co-cultured with kojic acid. On the other hand, when melanoma cells were treated with 800 μg / mL of black crystal roselle extract, the melanin content of melanoma cells was also significantly inhibited.

[0059] However, the above example is only one feasible embodiment and is not intended to limit the present invention.

[0060] [Third embodiment]

[0061] In this example, the elastin content analysis was used to evaluate whether the Roselle spatholobi extract had the potential to increase the elastin content. The method is as follows.

[0062] It should be noted that in order to understand whether the concentration of Roselle melanin extract used in this example is toxic to fibroblasts, a cell viability analysis was first performed, and it was confirmed that the concentrations of Roselle melanin extract used (100 μg / mL, 200 μg / mL, 400 μg / mL, and 800 μg / mL) were not cytotoxic (not shown).

[0063] Fibroblast CCD966SK (4×10 5 ) were seeded in a 24-well culture plate and cultured at 37°C for 24 hours. The cells were treated with different concentrations of Roselle extract for 48 hours, washed with PBS and treated with trypsin. The cell pellet was collected after centrifugation at 2000g for 10 minutes at 4°C, ultrasonically shaken, and centrifuged at 10000g for 10 minutes at 4°C to collect the supernatant. The elastin content was then analyzed using the Human Elastin ELISA Kit (ab239433).

[0064] like Figure 7As shown, when fibroblasts were treated with 200 μg / mL and 400 μg / mL of Roselle serrata extract, the elastin content of the fibroblasts was significantly increased, indicating that the Roselle serrata extract of the present invention has the potential to increase the elastin content.

[0065] However, the above example is only one feasible embodiment and is not intended to limit the present invention.

[0066] [Fourth embodiment]

[0067] In this example, the anti-UV ability of the Roselle spatholobi extract was further evaluated by UVA protection test, and the method is as follows.

[0068] Fibroblasts CCD966SK (8×10 5 ) were seeded in a 96-well culture plate and cultured at 37°C for 24 hours. The cells were treated with different concentrations of Roselle serrata extract for 24 hours, washed with PBS, and then added with 100 μl MEM-α (without phenol red). They were then placed in a VILBER-365 and incubated at 12 J / cm 2 After 60 minutes of irradiation, the cells were cultured for another 24 hours and the cell viability was analyzed by MTT assay.

[0069] like Figure 8 As shown, when fibroblasts were irradiated with UVA, the cell survival rate was significantly reduced to about 50% of the cell survival rate of the control group without UVA irradiation. However, when fibroblasts were treated with 100 μg / mL, 200 μg / mL, 400 μg / mL, and 800 μg / mL of Roselle melanogaster extract, the cell survival rate was significantly increased.

[0070] However, the above example is only one feasible embodiment and is not intended to limit the present invention.

[0071] [Beneficial Effects of Embodiments]

[0072] One of the beneficial effects of the present invention is that the black crystal roselle extraction method, the extract using the extraction method and the use thereof provided by the present invention can improve the active substance components contained in the black crystal roselle extract through the technical scheme of "step S1: providing dried black crystal roselle as an extraction raw material; step S2: finely grinding the extraction raw material; step S3: mixing the finely ground extraction raw material and the extraction solvent for primary extraction; step S4: ultrasonically extracting the black crystal roselle primary extract; filtering the black crystal roselle extract".

[0073] Furthermore, the black crystal roselle extract prepared by the above technical solution of the present invention exhibits beneficial effects such as inhibiting tyrosinase activity, reducing melanin content, increasing elastin content, and protecting cells from damage caused by UVA. That is, the black crystal roselle extract prepared by the black crystal roselle extraction method of the present invention has beneficial effects on skin care.

[0074] The contents disclosed above are only preferred feasible embodiments of the present invention, and are not intended to limit the protection scope of the claims of the present invention. Therefore, all equivalent technical changes made using the contents of the present invention description and drawings are included in the protection scope of the claims of the present invention.

Claims

1. A method for extracting black crystal roselle, characterized in that: The method for extracting black crystal Roselle comprises: Step S1: providing dried black crystal Roselle as an extraction raw material; Step S2: finely pulverizing the extracted raw material; Step S3: mixing the finely pulverized extraction raw material with an extraction solvent for primary extraction; Step S4: subjecting the Roselle rubrum extract to ultrasonic extraction; and Step S5: Filter and collect the black crystal roselle extract.

2. The method for extracting Roselle Black Crystal according to claim 1, characterized in that: The moisture content of the dried black crystal Roselle is less than 10%.

3. The method for extracting Roselle Black Crystal according to claim 1, characterized in that: The extraction solvent is water.

4. The method for extracting Roselle Black Crystal according to claim 3, characterized in that: The ratio of the finely pulverized extraction raw material to the water is 1:

15.

5. The method for extracting Roselle Black Crystal according to claim 4, characterized in that: The initial extraction is performed at a temperature of 60° C. for 15 to 60 minutes.

6. The method for extracting Roselle Black Crystal according to claim 1, characterized in that: The ultrasonic extraction is performed under ultrasonic conditions of 200W power and 40KHz frequency.

7. The method for extracting Roselle Black Crystal according to claim 6, characterized in that: The ultrasonic extraction is performed at a temperature of 60° C. for 15 to 60 minutes.

8. The method for extracting Roselle Black Crystal according to claim 1, characterized in that: The method for extracting black crystal Roselle further comprises: step S6: drying the black crystal Roselle extract.

9. A black crystal roselle extract, characterized in that: The black crystal roselle extract is prepared by the black crystal roselle extraction method according to claim 1.

10. A food composition, characterized in that The food composition comprises the Roselle melongena extract according to claim 9.

11. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the Roselle melanogaster extract according to claim 9.

12. A skin care cosmetic, characterized in that: The skin care cosmetic comprises the Roselle melanogaster extract according to claim 9.

13. Use of the black crystal roselle extract for preparing a pharmaceutical composition for skin care, characterized in that: The black crystal roselle extract is prepared by the black crystal roselle extraction method according to claim 1.

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