Preparation method and application of cleistocalyx operculatus flower extract with anti-aging effect

Abstract

The invention relates to a preparation method and application of a cleistocalyx operculatus flower extract with an anti-aging effect. Specifically, the invention discloses a preparation method of the cleistocalyx operculatus flower extract with the anti-aging effect, which comprises the following steps: extracting dried, crushed and sieved cleistocalyx operculatus flower by using water or an organic solvent according to a certain solid-to-liquid ratio at the temperature of 20-90 DEG C so as to extract the cleistocalyx operculatus flower extract with the anti-aging effect. According to the prepared cleistocalyx operculatus flower extract, the raw materials are natural single raw materials, and the components are purer, safer and more stable; the preparation method is simple, convenient, easy to operate, high in safety and remarkable in effect, and has wide market and application value in preparation of food or cosmetics with anti-oxidation and anti-skin-aging functions when being used as an active ingredient.

Description

Technical Field

[0001] The invention relates to the technical field of functional food and cosmetic raw material preparation, and more specifically, to a preparation method of an Echinops odoratus extract with anti-aging effects and application thereof. Background Art

[0002] Skin aging is affected by both internal and external factors. Elastin is located in the dermis of the skin in the skin structure and is composed of many monomers called tropoelastin. These monomers form a complex network structure through cross-linking. This cross-linked structure makes elastin highly elastic and can withstand multiple deformations without breaking easily, making the skin elastic and firm. Although its content is only 2-4% of the total skin protein, it is essential for the maintenance of skin structure. Reduced synthesis of elastin and fragmentation of elastic fibers can lead to skin aging. Elastase is an enzyme widely present in the body that can degrade elastin. Under normal conditions, the body has natural inhibitors of elastase to prevent elastin from being degraded. However, under pathological conditions such as aging of the body and long-term ultraviolet radiation, the concentration of elastase inhibitors decreases, or the activity of elastase increases, the dynamic balance between elastin and elastase is destroyed, and elastin is degraded. Skin tissue loses structural support and elasticity, causing the skin to sag and age.

[0003] In addition, free radicals are an important cause of skin aging. They not only damage telomeres, but also destroy collagen and elastic fibers in the skin, leading to skin sagging, increased wrinkles and other problems. Therefore, the exploration of substances with antioxidant free radical scavenging and elastase inhibitory activities is the key to the preparation of anti-skin aging active ingredients.

[0004] Cleistocalyx operculatus, scientific name: Cleistocalyx operculatus, is an evergreen tree of the Myrtaceae family. It is native to tropical Asia and is cultivated or wild in southern provinces of my country. It not only has high ornamental value but also has certain medicinal value. According to the "Lingnan Herbal Medicine Collection Record", "For medicinal use, old ones are better. Its flowers are bitter, cold in nature, clear away heat, detoxify, and eliminate food stagnation." In addition, Cleistocalyx operculatus also has anti-inflammatory, analgesic, and antibacterial effects, and can be used for trauma treatment and skin care.

[0005] At present, the application research of water lily in the field of anti-aging mainly has the following shortcomings:

[0006] 1. The ingredients are complex and use a variety of different active substances, which not only increases the cost, but also requires precise control of the proportion and ratio of each ingredient during production, high preparation requirements and complex operation.

[0007] 2. A mixture of multiple ingredients; single, natural ingredients are more popular with consumers.

[0008] 3. The synergistic effect of multiple ingredients makes it complicated to evaluate the specific contribution of each ingredient. In clinical trials and effect evaluations, it is difficult to determine the specific contribution of each ingredient to the final anti-aging effect.

[0009] Therefore, there is an urgent need in the art to develop a preparation method and application of an Eryngium truncatum extract with anti-aging effects. Summary of the invention

[0010] In view of the above problems, the purpose of the present invention is a preparation method of an Eryngium truncatum extract with anti-aging effect and its application.

[0011] In the first aspect of the present invention, a method for preparing an Ilex eryngii extract with anti-aging effects is provided, comprising extracting the dried, crushed and sieved Ilex eryngii with water or an organic solvent at a certain solid-liquid ratio at a temperature of 20-90°C, thereby extracting the Ilex eryngii extract with anti-aging effects.

[0012] In another preferred example, the Echinops truncatum includes Echinops truncatum flower buds.

[0013] In another preferred embodiment, the drying is carried out at a temperature of 25-90°C.

[0014] In another preferred embodiment, the sieving is performed using 50-100 mesh; preferably, 60-100 mesh; more preferably, 80 mesh.

[0015] In another preferred embodiment, the temperature is 25-90°C, preferably 30°C.

[0016] In another preferred embodiment, the organic solvent is selected from one or more combinations of the following groups:

[0017] Methanol, ethanol, propylene glycol, acetone, or ethyl acetate.

[0018] In another preferred embodiment, the concentration of the organic solvent is 50%-90%, preferably 65%-80%.

[0019] In another preferred embodiment, the solid-liquid ratio is 1:20 (g:ml)-1:50 (g:ml), preferably 1:30 (g:ml)-1:40 (g:ml), and more preferably 1:35 (g:ml).

[0020] In another preferred embodiment, the extraction method is selected from the following methods: ultrasonic-assisted extraction, flash extraction, or a combination thereof.

[0021] In another preferred embodiment, the ultrasonic assisted extraction adopts an ultrasonic frequency of 35 kHz, an ultrasonic temperature of 25° C., and an ultrasonic time of 40 min.

[0022] In another preferred embodiment, the flash extraction is performed using a flash extractor at 5000 rpm for 70 seconds.

[0023] In another preferred embodiment, the heating reflux extraction is performed using a heating reflux extractor at 65° C., and the extraction time is 150 min.

[0024] In another preferred embodiment, the extraction time is 2-200 min, preferably 4-150 min, and more preferably 40 min.

[0025] In another preferred example, the method further comprises: after the extraction, filtering or centrifuging to extract the supernatant, and concentrating and drying the supernatant to obtain the Eryngium wilfordii extract with anti-aging effects.

[0026] In another preferred embodiment, the centrifugal speed is 1000 rpm and the centrifugal time is 10 min.

[0027] In another preferred embodiment, the extracted process parameters are selected from any one or more combinations of the following groups:

[0028] (1) 80 mesh sieve, organic solvent is 80% ethanol, solid-liquid ratio is 1:30 (g:ml), ultrasonic assisted extraction, extraction temperature is 25 °C, extraction time is 40 min;

[0029] (2) 100 mesh sieve, the organic solvent is 65% ethanol, the solid-liquid ratio is 1:35 (g:ml), flash extraction is performed using a flash extractor, the extraction temperature is 30°C, and the extraction time is 4 min;

[0030] (3) 60 mesh sieve, the organic solvent is 80% methanol, the solid-liquid ratio is 1:40 (g:ml), heating reflux extraction is adopted, the extraction temperature is 90°C, and the extraction time is 150 min.

[0031] In a second aspect of the present invention, provided is an extract of Echeveria odorifera prepared by the method described in the first aspect of the present invention.

[0032] In another preferred embodiment, the DPPH free radical scavenging rate of the Echinops japonicus extract in an in vitro antioxidant activity test is ≥50% and the extract concentration is ≤1 mg / mL based on dry matter; and / or,

[0033] The inhibition rate of the extract of Echinops japonicus in an elastase inhibition test is ≥50% and the extract concentration is ≤5mg / mL based on dry matter.

[0034] In another preferred embodiment, the method for evaluating the biological activity of the Echinops odoratus extract comprises: an in vitro antioxidant test and / or an in vitro elastase inhibition test.

[0035] In another preferred embodiment, the method for determining the DPPH free radical scavenging rate in the in vitro antioxidant test is as follows:

[0036] Dilute the extract of Echeveria japonica to sample solutions with different concentration gradients; accurately weigh DPPH powder and dissolve it in anhydrous ethanol, and ultrasonicate for 15 minutes to prepare a DPPH solution with a concentration of 0.2mM; take 0.5mL sample solution and 1.5mL DPPH solution in a test tube AS as a sample tube; take 0.5mL distilled water and 1.5mL DPPH solution in a test tube Ab as a blank tube; take 0.5mL sample solution and 1.5mL anhydrous ethanol in a test tube Aj as a control tube; three test tubes in each sample are prepared at the same time and shaken evenly, and stand at 37℃ in the dark for 30 minutes; make three parallels and use an ELISA reader to measure the absorbance at 517nm; the DPPH free radical scavenging rate formula is as follows:

[0037] DPPH free radical scavenging rate (%) = [1-(As–Aj / Ab)] × 100

[0038] Among them, Ab is the absorbance of the blank group, As is the absorbance of the sample group, and Aj is the absorbance of the control group. IC50 is the concentration of the sample when the DPPH free radical scavenging rate reaches 50%, and vitamin C is used as the positive control.

[0039] In another preferred embodiment, the in vitro elastase inhibition is performed by diluting the above extract into different concentration gradients and mixing with Tris-HCl buffer, adding elastase solution, and then adding AAAPAN (N-succinyl-Ala Ala-Ala-p-nitroanilide), incubating at 25°C for 15 minutes, and measuring the absorbance; EGCG is used as a positive control.

[0040] In another preferred embodiment, the concentrations of the reagents used are: Tris-HCl buffer (50 Mmol / L, pH 8.0); elastase solution (60 mU / ml); AAAPAN (1 Mmol / L).

[0041] In another preferred embodiment, the method for determining the in vitro inhibitory activity of elastase of the Echinops odoratus extract is:

[0042] Samples of different concentrations (30 μL) were mixed with 85 μL of Tris-HCl buffer (50 Mmol / L, pH 8.0), and then 25 μL of elastase solution (60 mU / ml) was added and incubated at 25°C for 15 min. AAAPAN (25 μL, 1 M mol / L) was then added, and the absorbance was measured at 410 nm after 15 min. EGCG was used as a positive control. The calculation formula for elastase inhibitory activity is:

[0043] Inhibition rate (%) = [(AB) - (CD)] / (AB) × 100

[0044] Wherein A is the absorbance of the reaction solution without sample in the reaction system; B is the absorbance of the reaction solution without sample and enzyme in the reaction system; C is the absorbance of the reaction solution with sample and enzyme in the reaction system; D is the absorbance of the reaction solution without enzyme in the reaction system.

[0045] In the third aspect of the present invention, there is provided a use of the Echinops japonicus extract according to the second aspect of the present invention, wherein the Echinops japonicus extract is used for one or more combinations selected from the following groups:

[0046] (1) Used for preparing products with anti-oxidation and / or anti-skin aging effects;

[0047] (2) Used to prepare products with elastase inhibition function.

[0048] In another preferred embodiment, the product is selected from the following group:

[0049] Functional food, health product, cosmetic, or a combination thereof.

[0050] In a fourth aspect of the present invention, a cosmetic is provided, wherein the cosmetic contains the Eryngium wilfordii extract as described in the second aspect of the present invention.

[0051] In another preferred embodiment, the cosmetic further contains cosmetically acceptable auxiliary materials.

[0052] In a fifth aspect of the present invention, a food or health product is provided, wherein the food or health product contains the Eryngium wilfordii extract as described in the second aspect of the present invention.

[0053] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

[0054] Compared with the prior art, the present invention has the following technical effects:

[0055] (1) The extract of Echeveria sinensis with anti-aging effect prepared by the extraction method of the present invention is derived from a single plant source.

[0056] (2) The extraction method described in the present invention is simple, convenient and low-cost.

[0057] (3) The anti-aging mechanism of the water lily extract prepared by the extraction method of the present invention is mainly based on anti-oxidation and elastase inhibition. It can effectively inhibit the activity of elastase while removing free radicals in the skin and slowing down the skin aging process, thereby maintaining the elasticity and firmness of the skin.

[0058] (4) The preparation method of the present invention is simple, easy to operate, and has significant effects. It has broad market and application value when used as an active ingredient in the preparation of foods or cosmetics with antioxidant and anti-skin aging functions. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 A schematic diagram showing the scavenging effects of the water lily extracts and vitamin C (VC) prepared in Examples 1-3 of the present invention and the comparative example on 1,1-diphenyl-2-picrylhydrazyl (free radical) (DPPH) free radicals; wherein Examples 1-3 refer to water lily extracts 1-3 prepared based on Examples 1-3, respectively, and the comparative example refers to water lily extract 4 prepared based on comparative example 1.

[0060] Figure 2 A schematic diagram showing the in vitro inhibitory activity of the water lily extracts and vitamin C (VC) prepared in Examples 1-3 of the present invention and the comparative example on elastase; wherein Examples 1-3 refer to the water lily extracts 1-3 prepared based on Examples 1-3, respectively, and the comparative example refers to the water lily extract 4 prepared based on the comparative example 1. DETAILED DESCRIPTION

[0061] In order to make the purpose, technical solution, beneficial effects and significant progress of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, all the embodiments described are only partial embodiments of the present invention, not all embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0062] The flower buds of the water lily are dried, crushed, sieved, mixed with a solvent in a certain solid-liquid ratio and subjected to liquid-solid extraction at a certain temperature. The biological activity of the supernatant after solid-liquid separation of the extract is evaluated by a specific method, and the supernatant of the extract with high activity is subjected to rotary evaporation and drying to obtain the active extract.

[0063] Furthermore, the solid-to-liquid ratio of the Erythrina scabra to the solvent is 1:10-1:50 (g / mL).

[0064] Furthermore, the solvent is one or more combined solvents selected from water, methanol, ethanol, propylene glycol, acetone and ethyl acetate.

[0065] Furthermore, the biological activity evaluation method of the extract is a method of combining an in vitro antioxidant test with an in vitro elastase inhibition test.

[0066] Furthermore, the specific in vitro antioxidant test of the Echinops japonicus extract was evaluated by measuring the DPPH free radical scavenging rate:

[0067] Dilute the extract of Echeveria japonica to sample solutions with different concentration gradients; accurately weigh DPPH powder and dissolve it in anhydrous ethanol, and ultrasonicate for 15 minutes to prepare a DPPH solution with a concentration of 0.2mM; take 0.5mL sample solution and 1.5mL DPPH solution in a test tube AS as a sample tube; take 0.5mL distilled water and 1.5mL DPPH solution in a test tube Ab as a blank tube; take 0.5mL sample solution and 1.5mL anhydrous ethanol in a test tube Aj as a control tube; three test tubes in each sample are prepared at the same time and shaken evenly, and stand at 37℃ in the dark for 30 minutes; make three parallels and use an ELISA reader to measure the absorbance at 517nm; the DPPH free radical scavenging rate formula is as follows:

[0068] DPPH free radical scavenging rate (%) = [1-(As–Aj / Ab)] × 100

[0069] Among them, Ab is the absorbance of the blank group, As is the absorbance of the sample group, and Aj is the absorbance of the control group. IC50 is the concentration of the sample when the DPPH free radical scavenging rate reaches 50%, and the same concentration of vitamin C (VC) is used as the positive control.

[0070] Furthermore, the in vitro elastase inhibition is performed by diluting the above extract into different concentration gradients and mixing with Tris-HCl buffer, adding elastase solution, and then adding AAAPAN (N-succinyl-Ala Ala-Ala-p-nitroanilide), incubating at 25° C. for 15 minutes, and measuring the absorbance; the same concentration of epigallocatechin gallate (EGCG) is used as a positive control.

[0071] Furthermore, the concentrations of the reagents used were: Tris-HCl buffer (50 Mmol / L, pH 8.0); elastase solution (60 mU / ml); AAAPAN (1 Mmol / L).

[0072] Furthermore, the method for determining the in vitro inhibitory activity of elastase of the water lily extract is as follows: samples of different concentrations (30 μL) are mixed with 85 μL of Tris-HCl buffer (50 Mmol / L, pH 8.0), and then 25 μL of elastase solution (60 mU / ml) is added, and incubated at 25°C for 15 min. AAAPAN (25 μL, 1 M mol / L) is then added, and the absorbance is measured at 410 nm after 15 min. EGCG is used as a positive control. The calculation formula for elastase inhibitory activity is:

[0073] Inhibition rate (%) = [(AB) - (CD)] / (AB) × 100

[0074] Wherein A is the absorbance of the reaction solution without sample in the reaction system; B is the absorbance of the reaction solution without sample and enzyme in the reaction system; C is the absorbance of the reaction solution with sample and enzyme in the reaction system; D is the absorbance of the reaction solution without enzyme in the reaction system.

[0075] The extract of the water lily is calculated on a dry matter basis, and the DPPH free radical scavenging rate in the in vitro antioxidant activity test is selected to be ≥50% and the extract concentration is ≤1 mg / mL, and the inhibition rate in the elastase inhibition test is ≥50% and the extract concentration is ≤5 mg / mL to determine the extraction solvent and process parameters, thereby obtaining a method for preparing the water lily extract with antioxidant and anti-skin aging biological activities.

[0076] The raw materials and reagents involved in the present invention can all be purchased on the market.

[0077] Elastin has hydrophilicity and lipophilicity, can form a protective layer, reduce the sensitivity of the skin to environmental factors, and can increase the water content of the skin, having the effect of retaining water and moisturizing. Therefore, the extract of Echeveria odorifera prepared by the method of the present invention has good antioxidant and anti-skin aging effects, and it has important application value as an active ingredient in the preparation of functional foods, health products or cosmetics with antioxidant and anti-skin aging effects.

[0078] Preferably, the product comprises at least one of the following: functional food, health care product and cosmetics.

[0079] Example 1

[0080] 1.1 Preparation of Eryngium wilfordii extract:

[0081] After the flower buds of the water lily are dried, crushed and sieved, 80-mesh powder is taken and ultrasonic-assisted (35kHz, 25°C, 40min) extraction is carried out with 80% ethanol aqueous solution, the solid-liquid ratio of the extraction is 1:30g / mL, the extraction temperature is 25°C, the time is 40min, and the ultrasonic extraction is followed by centrifugation (1000rpm, 10min) for solid-liquid separation; the solid residue is extracted for a second time under the same conditions as above, and the supernatant is obtained by centrifugation after the extraction, the supernatants of the two extractions are combined and then evaporated and concentrated by a rotary evaporator, and the concentrated solution is freeze-dried to obtain the water lily extract 1, and the extract yield is 23.57% based on dry matter.

[0082] 1.2 DPPH free radical scavenging and elastase inhibition test, the specific process is as follows:

[0083] 1.2.1 Detection of DPPH free radical scavenging rate:

[0084] (1) diluting the above-mentioned Echeveria japonica extract into sample solutions with different concentration gradients (0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.4 mg / mL, 0.8 mg / mL, 1.2 mg / mL and 1.5 mg / mL);

[0085] (2) Accurately weigh DPPH powder and dissolve it in anhydrous ethanol, and ultrasonicate for 15 min to prepare a 0.2 mM DPPH solution;

[0086] (3) Take 0.5 mL of the above sample solution and 1.5 mL of DPPH solution in a test tube, marked as As, as a sample tube; take 0.5 mL of distilled water and 1.5 mL of DPPH solution in a test tube, marked as Ab, as a blank tube; take 0.5 mL of the above sample solution and 1.5 mL of anhydrous ethanol solution in a test tube, marked as Aj, as a control tube; three test tubes of each sample are simultaneously prepared and shaken evenly, and placed at 37°C in the dark for 30 min; make three parallels, and use a microplate reader to measure the absorbance at 517 nm; the DPPH free radical scavenging rate formula is as follows:

[0087] DPPH free radical scavenging rate (%) = [1-(As-Aj / Ab)] × 100

[0088] Among them, Ab is the absorbance of the blank group, As is the absorbance of the sample group, and Aj is the absorbance of the control group. IC50 is the concentration of the sample when the DPPH free radical scavenging rate reaches 50%, and the same concentration of vitamin C (VC) (reagents were purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd.) was used as the positive control.

[0089] The results are shown in Table 1. The DPPH scavenging IC50 of the extract 1 of Eryngium japonicum was 0.27±0.035 mg / mL.

[0090] 1.2.2 Detection of elastase inhibition rate:

[0091] (1) diluting the above-mentioned Echeveria japonica extract into sample solutions with different concentration gradients (0.4 mg / mL, 0.8 mg / mL, 1.5 mg / mL, 2 mg / mL and 3 mg / mL);

[0092] (2) Take 30 μL of the above sample solutions of different concentrations and mix them with 85 μL of Tris-HCl buffer (50 Mmol / L, pH 8.0), then add 25 μL of elastase solution (60 mU / ml) and incubate at 25°C for 15 min. Then add 25 μL of 1 M mol / L AAAPAN (N-succinyl-Ala Ala-Ala-p-nitroanilide) and incubate at 25°C for 15 min; measure the absorbance at 410 nm; use the same concentration of epigallocatechin gallate (EGCG) as a positive control (all reagents were purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd.).

[0093] The calculation formula of elastase inhibitory activity is:

[0094] Inhibition rate (%) = [(AB) - (CD)] / (AB) × 100

[0095] Wherein A is the absorbance of the reaction solution without sample in the reaction system; B is the absorbance of the reaction solution without sample and enzyme in the reaction system; C is the absorbance of the reaction solution with sample and enzyme in the reaction system; D is the absorbance of the reaction solution without enzyme in the reaction system.

[0096] The results are shown in Table 2. The IC50 of elastase inhibition of Echinops japonicus extract 1 was 1.75 mg / mL.

[0097] Example 2

[0098] Compared with Example 1, this embodiment is different in that:

[0099] After the flower buds of the water lily are dried, crushed and sieved, 100 mesh powder is taken and added with 65% ethanol aqueous solution at a solid-liquid ratio of 1:35 g / mL, and extracted by a flash extractor (5000 rpm, 70 s), the extraction temperature is 30°C, and the time is 4 min. After the flash extraction, the flower buds are filtered by rapid filter paper for solid-liquid separation to obtain solid residue and supernatant, the solid residue is extracted for the second time under the same conditions and solid-liquid separation is performed, the supernatants of the two extractions are combined, concentrated by rotary evaporation and freeze-dried to obtain the water lily extract 2, and the extract yield is 19.33% based on dry matter.

[0100] The results of DPPH free radical scavenging and elastase inhibition tests are shown in Tables 1 and 2, respectively. The DPPH scavenging IC50 of Echinops japonicus extract 2 is 0.07±0.019 mg / mL, and the elastase inhibition IC50 is 1.68 mg / mL.

[0101] Example 3

[0102] Compared with Example 1, this embodiment is different in that:

[0103] After the flower buds of the water lily are dried, crushed and sieved, 60 mesh powder is taken and added with 80% methanol aqueous solution at a solid-liquid ratio of 1:40 g / mL, and extraction is carried out using a heated reflux extractor (65°C) under 90°C water bath conditions. The extraction time is 150 minutes. After reflux extraction, it is filtered using rapid filter paper for solid-liquid separation to obtain solid residue and supernatant. The solid residue is extracted for a second time under the same conditions and solid-liquid separation is performed. The supernatants of the two extractions are combined and concentrated by rotary evaporation and freeze-dried to obtain the water lily extract 3, and the extraction yield based on dry matter is 23.74%.

[0104] The DPPH free radical scavenging and elastase inhibition tests were performed, and the results are shown in Tables 1 and 2, respectively. The DPPH scavenging IC50 of the water lily extract 3 was 0.33 mg / mL, and the elastase inhibition IC50 was 2.59 mg / mL.

[0105] Comparative Example 1

[0106] Compared with Example 1, this comparative example is different in that:

[0107] After the flower buds of the water lily are dried, crushed and sieved, 60 mesh powder is taken and dichloromethane is added at a solid-liquid ratio of 1:40 g / mL. The powder is extracted using a heated reflux extractor (40°C) under a 90°C water bath condition for 150 min. After reflux extraction, the powder is filtered using a rapid filter paper for solid-liquid separation to obtain a solid residue and a supernatant. The solid residue is extracted for a second time under the same conditions and solid-liquid separation is performed. The supernatants of the two extractions are combined and concentrated using rotary evaporation and freeze-dried to obtain the water lily extract 4. The extraction yield based on dry matter is 13.74%.

[0108] The results of DPPH free radical scavenging and elastase inhibition tests are shown in Tables 1 and 2, respectively. The DPPH scavenging IC50 of the water lily extract 4 is 0.63 mg / mL, and the elastase inhibition IC50 is 5.03 mg / mL.

[0109] When dichloromethane was used for extraction, the yield and activity of the extract were significantly lower than those in Example 1-3.

[0110] Experimental results analysis:

[0111] Based on the experimental process of the above-mentioned Examples 1-3 and the comparative example, the DPPH free radical scavenging and elastase inhibition test results of the extracted Echinops japonicus extracts 1-4 are shown in Table 1-2. The statistical results are shown in Table 1-2. Figure 1-2 As shown:

[0112] Table 1

[0113]

[0114] Table 2

[0115]

[0116]

[0117] 1) Based on the extract 1 obtained in Example 1, the extract yield was 23.57% based on dry matter; its DPPH scavenging IC50 was 0.27 mg / mL, and its elastase inhibition IC50 was 1.75 mg / mL;

[0118] 2) Based on the extract 2 obtained in Example 2, the extract yield was 19.33% based on dry matter; its DPPH scavenging IC50 was 0.07 mg / mL, and its elastase inhibition IC50 was 1.68 mg / mL;

[0119] 3) Based on the extract 3 of Echeveria japonica obtained in Example 3, the extract yield was 23.74% based on dry matter; its DPPH scavenging IC50 was 0.33 mg / mL, and its elastase inhibition IC50 was 2.59 mg / mL;

[0120] 4) Based on the extract 4 obtained in the comparative example, the extract yield was 13.74% based on dry matter; its DPPH scavenging IC50 was 0.63 mg / mL, and its elastase inhibition IC50 was 5.03 mg / mL;

[0121] 5) Positive control: VC's IC50 for DPPH scavenging is 0.03 mg / mL, and EGCG's IC50 for elastase inhibition is 2.03 mg / mL

[0122] The results show that the yield of the water lily extract, its DPPH scavenging activity and its elastase inhibitory activity are significantly lower than those of Examples 1-3 when extracted with dichloromethane. The extracts of Examples 1-3 have a high free radical scavenging activity. When the concentration is 0.4 mg / mL, the inhibition rate of the extract has reached about 90%, and then maintained a relatively stable level. In addition, the extracts of Examples 1-3 have a good inhibitory effect on elastase. At 1.5 to 3 mg / mL, the inhibition rate tends to be even better than the positive control EGCG. The experiment proves that the water lily extract prepared by this method can effectively scavenge free radicals in the body and inhibit elastase activity, thereby protecting cells from free radical damage, repairing muscle relaxation, keeping skin firm, and reducing wrinkles to achieve anti-aging effects.

[0123] The above describes the preferred embodiments of the present invention. It should be understood that the present invention is not limited to the above-mentioned specific embodiments, and the devices and structures that are not described in detail should be understood to be implemented in a common manner in the art; any technician familiar with the art can use the above-disclosed methods and technical contents to make many possible changes and modifications to the technical solutions of the present invention without departing from the scope of the technical solutions of the present invention, or modify them into equivalent embodiments of equivalent changes, which does not affect the essential content of the present invention. Therefore, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solutions of the present invention are still within the scope of protection of the technical solutions of the present invention.

Claims

1. A method for preparing an extract of Echinops odoratus with anti-aging effect, characterized in that: The method comprises extracting the dried, crushed and sieved water lily with water or an organic solvent at a certain solid-liquid ratio at a temperature of 20-90 DEG C, so as to extract the water lily extract with anti-aging effect.

2. The preparation method according to claim 1, characterized in that The organic solvent is selected from one or more combinations of the following groups: Methanol, ethanol, propylene glycol, acetone, or ethyl acetate.

3. The preparation method according to claim 1, characterized in that: The concentration of the organic solvent is 50%-90%, preferably 65%-80%.

4. The preparation method according to claim 1, characterized in that: The solid-liquid ratio is 1:20 (g:ml)-1:50 (g:ml), preferably 1:30 (g:ml)-1:40 (g:ml), and more preferably 1:35 (g:ml).

5. The preparation method according to claim 1, characterized in that: The extraction time is 2-200 min, preferably 4-150 min, and more preferably 40 min.

6. The extract of Echeveria odorata prepared by the method according to any one of claims 1 to 5.

7. The Eryngium wilfordii extract according to claim 6, characterized in that The DPPH free radical scavenging rate of the extract of the water lily is ≥50% and the extract concentration is ≤1 mg / mL in an in vitro antioxidant activity test based on dry matter; and / or, The inhibition rate of the extract of Echinops japonicus in an elastase inhibition test is ≥50% and the extract concentration is ≤5mg / mL based on dry matter.

8. The use of the Eryngium wilfordii extract according to any one of claims 6 to 7, characterized in that: The extract of Echeveria odorata is used for one or more combinations selected from the following groups: (1) Used for preparing products with anti-oxidation and / or anti-skin aging effects; (2) Used to prepare products with elastase inhibition function.

9. The use according to claim 8, characterized in that The product is selected from the group consisting of: Functional food, health product, cosmetic, or a combination thereof.

10. A cosmetic, characterized in that: Contains the Eryngium wilfordii extract according to claim 6.

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