Method for extracting anti-aging component camellia saponin A from camellia based on full-bionic targeted extraction technology and application

Abstract

The invention discloses a method for extracting an anti-aging component camellia saponin A from camellia on the basis of a full-bionic targeted extraction technology and application of the anti-aging component camellia saponin A. Efficient enrichment and structure protection of the camellia saponin A are achieved by simulating the whole process of decomposition and absorption of the oral cavity, the stomach and the intestinal tract of a human body. Specifically, the camellia flowers or leaves are pretreated through supercritical extraction, lipid and gel impurities which are contained in the camellia flowers or leaves and are prone to forming gelatinoids are removed, and convenience is provided for subsequent purification; the problems that the camellia saponin A is low in yield and easy to degrade are solved by applying a full-bionic enzymolysis technology, and meanwhile, impurities such as lipid, protein, tannin and sugar can be degraded through full-bionic extraction, so that convenience is provided for subsequent crystallization; and purifying by adopting a macroporous resin adsorption technology, and finally purifying by crystallization. The camellia saponin A obtained by the invention can better achieve the effects of improving wrinkles, repairing skin, brightening skin color and delaying senescence through multi-way and multi-azimuth anti-wrinkle, repairing and whitening effects, and can be widely applied to various types of cosmetics.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical and application field of Camellia sinensis deep processing, and relates to a method for extracting Camelliaside A from Camellia sinensis and application thereof in anti-aging; in particular, the present application relates to a method for extracting anti-aging component Camelliaside A from Camellia sinensis based on full-bionic targeted extraction technology and application thereof. The Camelliaside A obtained by the method has the effects of anti-wrinkle, repair and whitening, and can better achieve the effects of improving wrinkles, repairing skin, lightening skin color and delaying aging, and can be widely applied to cosmetics as an effective raw material. BACKGROUND

[0002] Camellia sinensis is a long-lived oil woody plant of the Camellia family, and the distribution of plants in the Camellia family is mainly concentrated in China. With the rise of the Chinese cosmetic industry, "Chinese ingredients" are increasingly favored by consumers. Camellia flowers, as a common ornamental plant in China, contain various effective components in the flowers, such as polyphenols, triterpenes, alkaloids and the like; the leaves of Camellia sinensis contain rich flavonoids, polyphenols and triterpenes. Camellia plants are rich in various active ingredients, which are important effective additives for daily chemical products, and have various effects such as antioxidant, antibacterial, anti-allergic, anti-inflammatory and promoting wound healing.

[0003] Camelliaside A is a high-activity triterpene saponin compound with the effects of repair, anti-wrinkle, antioxidant, anti-inflammatory and whitening, and is widely used in the fields of cosmetics, medicine and the like. At present, the industrialized extraction process still faces technical bottlenecks such as low extraction efficiency, easy damage of active ingredients, high solvent residue and the like. The preparation method of Camelliaside A includes physical conversion method, chemical conversion method and biological conversion method. The physical conversion method includes microwave treatment method, heating extraction method, and microwave / ultrasonic assisted method, which can shorten the extraction time, but the local high temperature causes the degradation of heat-sensitive components, and the bioactivity retention rate is low; the solvent extraction method uses ethanol or methanol for extraction, and has the problem of poor selectivity of saponin A (yield ≤1.0%) (such as CN 107280993A), and needs multi-step purification (silica gel column chromatography→HPLC preparation) in the subsequent process, and the production cycle is long. The chemical conversion method includes acid hydrolysis method, alkali hydrolysis method and the like, and the acid and alkali reagents are not environmentally friendly and have high requirements for the operating environment and personnel; the biological conversion method includes enzymatic hydrolysis method and the like, and the existing technology of biological enzymatic hydrolysis mainly uses single cellulase for treatment, which cannot effectively break the complex structure of Camellia flower cell wall (containing lignin-pectin covalent cross-linking network), and the enzymatic hydrolysis efficiency is low; the saponin A and non-target saponin (such as saponin B / C Camelliaside B / C) coexist in the traditional enzymatic hydrolysis product, which increases the cost of subsequent separation by 2-3 times.

[0004] This invention innovatively combines supercritical extraction technology with targeted extraction technology based on a biomimetic oral-gastrointestinal absorption mechanism for the first time. Supercritical extraction technology is used to remove lipid and gel components from camellia; fully biomimetic extraction technology is used to decompose lipid, protein, polysaccharide, and gelatin components in crude camellia extracts to form colloidal substances; and targeted extraction technology based on a biomimetic oral-gastrointestinal absorption mechanism is used to remove the sugar rings of camellia saponin A parent ring structure (such as Camelliaside A / B / C) and to increase the content of camellia saponin A through purification and enrichment. Patent CN17883347A describes a method for preparing ginsenosides using multiple enzymes. After enzymatic hydrolysis, at least six rare ginsenosides are added, but the patent does not mention the conversion rate before and after enzymatic hydrolysis. The purity of camellia saponin A prepared by the present invention is more than three times higher than that of traditional processes, thereby significantly improving anti-aging, repair, and whitening effects. The present invention provides a reference for the deep processing of camellia. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention provides a method and application for extracting camellia saponin A, an anti-aging ingredient, from camellia using a fully biomimetic targeted extraction technology. The extract obtained using the present method has a high content of active ingredients, is environmentally friendly, and has high extraction efficiency. Furthermore, the method is energy-efficient, time-efficient, and simple to operate, making it suitable for industrial production.

[0006] The object of the present invention is achieved through the following technical solutions:

[0007] The present invention provides a method for extracting camellia saponin A from camellia, comprising the following steps:

[0008] A1. Take camellia flowers or leaf powder and perform supercritical extraction;

[0009] The powders obtained by extraction of A2 and A1 were ultrasonically treated with anhydrous ethanol, stirred and extracted, filtered to obtain a camellia extract, and concentrated and dried to obtain a crude extract of camellia saponin A;

[0010] A3, adding the crude extract of camellia saponin A to water, adding α-amylase to simulate the oral environment, and reacting at 30-50° C. with the pH range controlled between 6.0-7.0 for 0.5-2 h; then adding an acidic protease module to simulate the gastric environment, and reacting at 40-50° C. with the pH range controlled between 1.5-3.5 for 0.5-2 h; then adding trypsin and lipase to simulate the intestinal environment, and reacting at 40-50° C. with the pH range controlled between 6.5-7.5 for 0.5-2 h; then heating to inactivate the enzymes, and filtering to obtain a biomimetic extract of camellia saponin A;

[0011] A4, the Camellia saponin A biomimetic extract is subjected to segmental purification by macroporous resin, and Camellia saponin A eluate rich in Camellia saponin A is collected;

[0012] A5, the Camellia saponin A eluate is concentrated to a solid content of 10-16%, then diluted with anhydrous ethanol to a solid content of 5-8%, and then allowed to stand at 2-8°C for 10-15 hours, and then filtered; and the filtrate is concentrated and dried to obtain Camellia saponin A.

[0013] As an embodiment, in step A1, the supercritical extraction is performed at an extraction pressure of 15-45 MPa, an extraction temperature of 45-55°C, and an extraction time of 30 min-120 min. Preferably, the supercritical extraction is performed at an extraction pressure of 15-40 MPa.

[0014] As an embodiment, in step A1, the Camellia flower or leaf powder is prepared by crushing Camellia flower or leaf to a size of 100-160 mesh. The crushed Camellia flower or leaf is placed in a supercritical extraction device, and subjected to degreasing and degelation at an extraction pressure of 15-45 MPa, an extraction temperature of 45-55°C, and an extraction time of 30 min-120 min. After the extraction is completed, the Camellia flower or leaf is removed, and Camellia flower or leaf powder A is obtained.

[0015] As an embodiment, in step A2, the powder obtained by extraction in A1 is added to 30-80% volume fraction anhydrous ethanol in a mass ratio of 10-30:1, and the solution temperature is controlled below 40°C, and then subjected to ultrasonic treatment.

[0016] As an embodiment, the ultrasonic treatment is performed at an ultrasonic power of 500 W-3000 W and an ultrasonic time of 10 min-60 min.

[0017] As an embodiment, in step A2, the solution temperature of the stirring extraction is controlled at 40°C-70°C, and the stirring extraction time is 1 h-4 h. After the extraction is completed, the extraction solution is allowed to cool to room temperature, filtered to obtain Camellia extract, and concentrated and dried to obtain Camellia saponin A crude extract.

[0018] As an embodiment, step A3 includes at least one of the following technical features:

[0019] B1, the Camellia saponin A crude extract is added to water in a mass ratio of 10-50:1;

[0020] B2, the α-amylase is added in an amount of 0.5-3% of the mass of the Camellia saponin A crude extract;

[0021] B3, the acid protease is added in an amount of 0.5-3% of the mass of the Camellia saponin A crude extract;

[0022] B4, the added amount of the trypsin is 0.5-3% of the mass of the crude sasanqua saponin A extract;

[0023] B5, the added amount of the lipase is 0.5-3% of the mass of the crude sasanqua saponin A extract;

[0024] B6, the temperature for heating and killing the enzyme is 80-100℃, and the time for killing the enzyme is 5-20 minutes.

[0025] As an embodiment, in step A4, the macroporous resin is one of YKDH-5, D-101, AB-8, X-5, and NKA-9; preferably, the macroporous resin is D-101 or AB-8; most preferably, the macroporous resin is D-101.

[0026] And / or, in the segment purification, the loading amount of the sasanqua saponin A biomimetic extract is 1-3BV, BV is the bed volume of the resin; then 1-3BV water is used for elution, and the eluate is discarded; then 30-80% ethanol is used for segment elution, 1-6BV is used for elution, and the eluate rich in sasanqua saponin A is collected.

[0027] As an embodiment of the present application, in the segment purification, the loading amount of the sasanqua saponin A biomimetic extract is 1-3BV, BV is the bed volume of the resin; then 1-3BV water is used for elution, and the eluate is discarded; then 1-2BV 20%-40% ethanol is used for elution, and the eluate is discarded; finally, 40%-80% ethanol is used for elution, 2-4BV is used for elution, and the eluate rich in sasanqua saponin A is collected. Preferably, 2BV 30%-40% ethanol is used for elution, and the eluate is discarded; finally, 40%-70% ethanol is used for elution, 2-4BV is used for elution. More preferably, 2BV 30%-40% ethanol is used for elution, and the eluate is discarded; finally, 60%-70% ethanol is used for elution, 2-4BV is used for elution.

[0028] As an embodiment of the present application, preferably, the sasanqua saponin A eluate is concentrated to a solid content of 10-16%, then diluted with anhydrous ethanol to a solid content of 6-8%, and then placed at 2-8℃ for 10-15h, and then filtered; the filtrate is concentrated and dried to obtain sasanqua saponin A.

[0029] The sasanqua saponin A extract obtained by the method of the present application and the application thereof in cosmetics all belong to the protection scope of the present application. The sasanqua saponin A extract in the cosmetics is an anti-aging component. The cosmetics can be anti-wrinkle and repair cosmetics.

[0030] Compared with the prior art, the present application has the following beneficial effects:

[0031] (1) The application first innovatively combines supercritical extraction technology with full-bionic extraction technology. The application removes lipids and gel components in camellia flowers by supercritical extraction technology, improves the extraction rate of camellia saponin A, and improves the effect of macroporous resin purification and camellia saponin A crystallization. The application solves the problems of low yield and easy degradation of camellia saponin A by full-bionic extraction, and improves the effect of macroporous resin purification and camellia saponin crystallization by degrading lipids, proteins, tannins, sugars and other impurities. The content of camellia saponin A prepared by the application is more than 3 times higher than that by traditional process.

[0032] (2) The application first applies macroporous resin step-by-step purification technology to the purification of camellia saponin A.

[0033] (3) The camellia saponin A prepared by the application greatly improves the skin care effect, especially the anti-wrinkle and repair effect, which is more than doubled. BRIEF DESCRIPTION OF DRAWINGS

[0034] Other features, objects and advantages of the application will become more apparent from the following detailed description of non-limiting embodiments, with reference to the accompanying drawings:

[0035] Figure 1 is a schematic diagram of the sample of the comparative example in the recrystallization process; wherein, 1: comparative example 1; 2: comparative example 2; 3: comparative example 6; 4: comparative example 7; 5: comparative example 8;

[0036] Figure 2 is a chromatogram of camellia saponin A of example 7 and its comparison with the standard product;

[0037] Figure 3 is a mass spectrum of camellia saponin A of example 7;

[0038] Figure 4 is a schematic diagram of the structure of camellia saponin A. DETAILED DESCRIPTION

[0039] The application will be described in detail below with reference to the examples. The following examples will help those skilled in the art to further understand the application, but do not limit the application in any form. It should be noted that those skilled in the art can make several adjustments and improvements without departing from the concept of the application. These are within the scope of protection of the application. The extraction rate (%) = mass of extract (g) / mass of raw material (g), in this case, the identification method of camellia saponin A is adjusted according to the analysis method in

[0031] paragraph of CN 107280993A specification, and the specific test method is as follows.

[0040] Chromatographic conditions: ZORBAX-SB-C18 column (4.6 mm x 150 mm, 2.7 μm), column temperature: (30 ± 5) °C; mobile phase: acetonitrile (A) - 0.01% glacial acetic acid water (B) gradient elution, see Table 1, flow rate of 1 mL / min; injection volume 10 μL, see Figure 2 .

[0041] Table 1 Gradient elution table

[0042] Time Acetonitrile (%) 0.01 % Glacial Acetic Acid Water (%) 0.00 10 90 8.00 50 50 12.00 95 5 16.00 95 5 16.01 10 90

[0043] Mass spectrometry conditions: TOF MS-IDA-MS / MS mode, electrospray ion source (ESI), positive and negative ion mode scanning, mass scan range m / z 100-1000. Curtain gas (CUR) 35 psi, nebulizing gas (GS1) 55 psi, auxiliary gas (GS2) 55 psi, electrospray voltage (ISVF) 5500 V, ion source temperature (TEM) 550 °C, de-clustering voltage (DP) 80 V; TOF-MS mode collision voltage 10 V, IDA-MS / MS collision voltage 35 V, collision voltage difference 15 V, see Appendix 2. Figure 3 .

[0044] Example 1

[0045] A1, take 100 g of camellia flowers, crush through a 100 mesh sieve, and place in a supercritical extraction device, under extraction pressure 30 MPa, extraction temperature 45 °C, extraction time 60 min, defat and degelatinize, after extraction, take out the camellia flowers, to obtain camellia flower powder A;

[0046] A2, take the supercritical camellia flower powder, add 30 times the volume of an ethanol water solution, with an ethanol volume content of 80%; control the ethanol water solution of the sample to below 40 °C, then ultrasonic for 60 min;

[0047] A3, control the solution in step A2 to 70 °C, and stir extract for 1 h; after extraction is complete, wait for the extraction liquid temperature to drop to room temperature, filter to obtain camellia flower extract, concentrate and dry to obtain camellia saponin A crude extract, with an extraction rate of 5.45% and a purity of 8.12%;

[0048] A4, the crude extract of camellia saponin A obtained in step A3 is added to water in a mass of 50 times the volume, 3% of the mass of dry substance A of α-amylase is added to simulate the oral environment, and reacted at 50°C and pH 7.0 for 2h; 3% of acid protease is then added to simulate the gastric environment module, and reacted at 50°C and pH 3.5 for 2h; 3% of trypsin and 3% of lipase are then added to simulate the intestinal environment module, and reacted at 50°C and pH 7.5 for 2h; then heated to 90°C to inactivate the enzyme for 10 minutes, and filtered to obtain a camellia saponin A solution (i.e. a biomimetic extract).

[0049] A5, the biomimetic extract obtained in step A4 is purified by macroporous resin D-101, the loading amount is 1-3BV (BV is the bed volume of the resin), then 2BV of water is used to elute and discard the eluate; 2BV of 20% ethanol is then used to elute and discard the eluate, and finally 4BV of 80% ethanol by volume is used to elute, and the eluate rich in camellia saponin A is collected.

[0050] A6, the above concentrated solution is concentrated to a solid content of 10%, then diluted with anhydrous ethanol to a solid content of 5%, and then left still at 4°C for 12h, then filtered, and finally the filtered concentrated solution is concentrated and dried to obtain camellia saponin A, with an extraction rate of 0.64% and a purity of 54.16%.

[0051] Example 2

[0052] A1, 100g of camellia flowers are pulverized through a 100-mesh sieve and placed in a supercritical extraction device, and defatted and degelatinized at an extraction pressure of 15MPa, an extraction temperature of 50°C, and an extraction time of 120min; after the extraction is completed, the camellia flowers are removed, i.e. camellia flower powder A is obtained;

[0053] A2, the supercritically extracted camellia flower powder is pulverized, and 10 times the amount of an ethanol aqueous solution is added to the supercritically extracted camellia flower powder, wherein the ethanol content is 30%; the ethanol aqueous solution of the sample is then controlled to be below 40°C, and then ultrasonicated for 30min;

[0054] A3, the solution in step A2 is controlled to be at 70°C, and extracted by stirring for 1.5h; after the extraction is completed, the temperature of the extract is allowed to decrease to room temperature, and then filtered to obtain a camellia extract, which is concentrated and dried to obtain a crude extract of camellia saponin A, with an extraction rate of 5.21% and a purity of 7.62%;

[0055] A4, the crude extract of camellia saponin A obtained in step A3 is added to water in a mass-volume ratio of 20 times, 0.5% of α-amylase is added to simulate the oral environment, and the reaction is carried out at 50°C and pH 7.0 for 2h; then 0.5% of acid protease is added to simulate the stomach environment module, and the reaction is carried out at 50°C and pH 3.5 for 2h; then 0.5% of trypsin and 0.5% of lipase are added to simulate the intestinal environment module, and the reaction is carried out at 50°C and pH 7.5 for 2h; then heat to 90°C, and kill the enzyme for 10 minutes. Filter to obtain a camellia saponin A solution.

[0056] A5, the biomimetic extract obtained in step A4 is purified by macroporous resin YKDH-5, the loading amount is 2BV (BV is the bed volume of the resin), then 2BV of water is used for elution and the eluate is discarded; then 2BV of 40% ethanol is used for elution and the eluate is discarded, and then 70% of the extract fraction is eluted with 2BV of anhydrous ethanol, and the eluate rich in camellia saponin A is collected.

[0057] A6, the above concentrated solution is concentrated to a solid content of 16%, then diluted with anhydrous ethanol to a solid content of 5%, and then left still in an environment of 8°C for 12h, then filtered, and finally the filtered concentrated solution is concentrated and dried to obtain camellia saponin A, with an extraction rate of 0.86% and a purity of 43.18%.

[0058] Example 3

[0059] A1, take camellia 100g, crush through 100 mesh sieve, and place in a supercritical extraction device, and defat and degel at an extraction pressure of 25MPa, an extraction temperature of 50°C, and an extraction time of 60min. After the extraction is completed, the camellia is taken out, and camellia powder A is obtained;

[0060] A2, take the supercritical camellia powder, crush, and add 25 times of an ethanol aqueous solution, wherein the ethanol content is 60%; control the ethanol aqueous solution of the sample below 40°C, and then ultrasonic for 40min;

[0061] A3, control the solution in step A2 at 40°C, and stir and extract for 4h; after the extraction is completed, wait for the temperature of the extract to drop to room temperature, filter to obtain a camellia extract, and concentrate and dry to obtain a crude extract of camellia saponin A, with an extraction rate of 5.53% and a purity of 8.34%;

[0062] A4, the crude extract of camellia saponin A obtained in step A3 is added to water in a mass of 50 times the volume, 1% of the mass of dry substance A of α-amylase is added to simulate the oral environment, and reacted at 50°C and pH 7.0 for 2h; 1% of acid protease is then added to simulate the gastric environment module, and reacted at 50°C and pH 3.5 for 2h; 1% of trypsin and 1% of lipase are then added to simulate the intestinal environment module, and reacted at 50°C and pH 7.5 for 2h; and then heated to 90°C to inactivate the enzyme for 10 minutes, and filtered to obtain a camellia saponin A solution.

[0063] A5, the biomimetic extract obtained in step A4 is purified by AB-8 macroporous resin, the loading amount is 1BV (BV is the bed volume of the resin), and then 3BV of water is used for elution, and the eluate is discarded; 2BV of 30% ethanol is then used for elution, and the eluate is discarded, and 40% of the extract fraction is eluted with 4BV of anhydrous ethanol, and the camellia saponin A eluate is collected.

[0064] A6, the above concentrated solution is concentrated to a solid content of 12%, then diluted with anhydrous ethanol to a solid content of 6%, and then left still at 2°C for 12h, and then filtered, and finally the filtered concentrated solution is concentrated and dried to obtain camellia saponin A, with an extraction rate of 0.61% and a purity of 62.13%.

[0065] Example 4

[0066] A1, 100g of camellia leaves are crushed to pass through a 100-mesh sieve, and placed in a supercritical extraction device, and defatted and degelatinized at an extraction pressure of 30MPa, an extraction temperature of 55°C, and an extraction time of 30min, and after the extraction is completed, the camellia leaves are removed, and camellia leaf powder A is obtained;

[0067] A2, the supercritically extracted camellia leaf powder is crushed, and 20 times the amount of an ethanol aqueous solution is added, wherein the ethanol content is 40%, and the ethanol aqueous solution of the sample is then controlled to be below 40°C, and then ultrasonicated for 50min;

[0068] A3, the solution in step A2 is controlled to be at 40°C, and extracted by stirring for 3h; after the extraction is completed, the temperature of the extract is allowed to decrease to room temperature, and then filtered to obtain a camellia leaf extract, which is concentrated and dried to obtain a crude extract of camellia saponin A, with an extraction rate of 5.16% and a purity of 9.28%;

[0069] A4, the crude extract of camellia saponin A obtained in step A3 is added to water in a mass of 50 times the volume, 1.5% of the mass of dry substance A of α-amylase is added to simulate the oral environment, and reacted at 50°C and pH 7.0 for 2h; 1.5% of acid protease is added to simulate the gastric environment module, and reacted at 50°C and pH 3.5 for 2h; 1.5% of trypsin and 1.5% of lipase are added to simulate the intestinal environment module, and reacted at 50°C and pH 7.5 for 2h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and filtration is performed to obtain a camellia saponin A solution.

[0070] A5, the biomimetic extract obtained in step A4 is purified by macroporous resin X-5, the loading amount is 2BV (BV is the bed volume of the resin), then 2BV of water is used for elution, and the eluate is discarded; then 2BV of 20% ethanol is used for elution, and the eluate is discarded, then 60% of the extract fraction is anhydrous ethanol, and elution is performed for 4BV, and the eluate rich in camellia saponin A is collected.

[0071] A6, the above concentrated solution is concentrated to a solid content of 10%, then diluted with anhydrous ethanol to a solid content of 8%, and then left still in an environment of 5°C for 12h, then filtered, and finally the filtered concentrated solution is concentrated and dried to obtain camellia saponin A, the extraction rate is 0.92%, and the purity is 41.28%.

[0072] Example 5

[0073] A1, 100g of camellia leaves are crushed to pass through a 100-mesh sieve, and placed in a supercritical extraction device, and defatted and degelatinized at an extraction pressure of 40MPa, an extraction temperature of 45°C, and an extraction time of 80min, and after the extraction is completed, the camellia leaves are taken out, and camellia leaf powder A is obtained;

[0074] A2, the supercritically extracted camellia leaf powder is crushed, and 30 times the amount of an ethanol aqueous solution is added, wherein the ethanol content is 60%; the ethanol aqueous solution of the sample is controlled to be below 40°C, and then ultrasonicated for 60min;

[0075] A3, the solution in step A2 is controlled to be at 50°C, and extracted by stirring for 2h; after the extraction is completed, the temperature of the extract is lowered to room temperature, filtration is performed to obtain a camellia leaf extract, and concentrated and dried to obtain a crude extract of camellia saponin A, the extraction rate is 5.53%, and the purity is 8.63%;

[0076] A4, the crude extract of camellia saponin A obtained in step A3 is added to water in a mass of 40 times the volume, 3% of the mass of dry substance A of α-amylase is added to simulate the oral environment, and reacted at 50°C and pH 7.0 for 2h; 3% of acid protease is then added to simulate the gastric environment module, and reacted at 50°C and pH 3.5 for 2h; 3% of trypsin and 3% of lipase are then added to simulate the intestinal environment module, and reacted at 50°C and pH 7.5 for 2h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and filtration is performed to obtain a camellia saponin A solution.

[0077] A5, the biomimetic extract obtained in step A4 is purified by macroporous resin X-5, the loading amount is 2BV (BV is the bed volume of the resin), then 2BV of water is used for elution and the eluate is discarded; then 2BV of 30% ethanol is used for elution and the eluate is discarded, and then 80% of the extract fraction is eluted with 2BV of anhydrous ethanol, and the eluate rich in camellia saponin A is collected.

[0078] A6, the above concentrated solution is concentrated to a solid content of 16%, then diluted with anhydrous ethanol to a solid content of 8%, and then left still at 4°C for 12h, then filtered, and finally the filtered concentrated solution is concentrated and dried to obtain camellia saponin A, the extraction rate is 0.72%, and the purity is 48.12%.

[0079] Example 6

[0080] A1, 100g of camellia leaves are crushed to pass through a 100-mesh sieve, and placed in a supercritical extraction device, and defatted and degelatinized at an extraction pressure of 30MPa, an extraction temperature of 45°C, and an extraction time of 100min, and after the extraction is completed, the camellia leaves are removed, and camellia leaf powder A is obtained;

[0081] A2, the supercritically extracted camellia leaf powder is crushed, and 20 times the amount of an ethanol aqueous solution is added, wherein the ethanol content is 60%, and the ethanol aqueous solution of the sample is controlled to be below 40°C, and then ultrasonicated for 60min;

[0082] A3, the solution in step A2 is controlled to be at 60°C, and extracted by stirring for 3h; after the extraction is completed, the temperature of the extract is lowered to room temperature, and filtration is performed to obtain a camellia leaf extract, which is concentrated and dried to obtain a crude extract of camellia saponin A, the extraction rate is 5.16%, and the purity is 9.24%;

[0083] A4, the crude extract of camellia saponin A obtained in step A3 is added to water in a mass-volume ratio of 20 times, 1.5% of the mass of dry substance A of α-amylase is added to simulate the oral environment, and reacted at 50°C and pH 7.0 for 2h; 1.5% of acid protease is added to simulate the gastric environment module, and reacted at 50°C and pH 3.5 for 2h; 1.5% of trypsin and 1.5% of lipase are added to simulate the intestinal environment module, and reacted at 50°C and pH 7.5 for 2h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and filtration is performed to obtain a camellia saponin A solution.

[0084] A5, the biomimetic extract obtained in step A4 is purified by macroporous resin NKA-9, the loading amount is 2BV (BV is the bed volume of the resin), then 2BV of water is used for elution and the eluate is discarded; 2BV of 20% ethanol is used for elution and the eluate is discarded, and 2BV of 70% ethanol is used for elution, and the camellia saponin A eluate is collected.

[0085] A6, the above concentrated solution is concentrated to a solid content of 16%, then diluted with anhydrous ethanol to a solid content of 8%, and then placed in an environment at 4°C for 12h, then filtered, and finally the filtered concentrated solution is concentrated and dried to obtain camellia saponin A, the extraction rate is 0.84%, and the purity is 45.14%.

[0086] Example 7

[0087] A1, 100g of camellia flowers is crushed to pass through a 100-mesh sieve, and placed in a supercritical extraction device, and defatted and degelatinized at an extraction pressure of 25MPa, an extraction temperature of 52°C, and an extraction time of 45min, and then the camellia flowers are taken out, and camellia flower powder A is obtained;

[0088] A2, the supercritical camellia flower powder is crushed, and 20 times of an ethanol aqueous solution is added to the supercritical camellia flower powder, wherein the ethanol content is 60%; the ethanol aqueous solution of the sample is controlled below 40°C, and then ultrasonic treatment is performed for 60min;

[0089] A3, the solution in step A2 is controlled at 50°C, and stirred for extraction for 2h; after the extraction is completed, the temperature of the extract is lowered to room temperature, filtration is performed to obtain a camellia extract, and concentration and drying are performed to obtain a crude extract of camellia saponin A, the extraction rate is 5.52%, and the purity is 8.55%;

[0090] A4, the crude extract of camellia saponin A obtained in step A3 is added to water in a mass-volume ratio of 20 times, 2% of the mass of dry substance A of α-amylase is added to simulate the oral environment, and reacted at 50°C and pH 7.0 for 2h; 2% of acid protease is then added to simulate the gastric environment module, and reacted at 50°C and pH 3.5 for 2h; 2% of trypsin and 2% of lipase are then added to simulate the intestinal environment module, and reacted at 50°C and pH 7.5 for 2h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and filtration is performed to obtain a camellia saponin A solution.

[0091] A5, the biomimetic extract obtained in step A4 is purified by macroporous resin D-101, the loading amount is 2BV (BV is the bed volume of the resin), then 2BV of water is used for elution and the eluate is discarded; then 2BV of 30% ethanol is used for elution and the eluate is discarded, and then 2BV of 60% ethanol is used for elution, and the eluate rich in camellia saponin A is collected.

[0092] A6, the above concentrated solution is concentrated to a solid content of 16%, then diluted with anhydrous ethanol to a solid content of 8%, and then left still in an environment of 2°C for 12h, then filtered, and finally the filtered concentrated solution is concentrated and dried to obtain camellia saponin A (see Figure 2 , Figure, Figure 3 , Figure 4 ), the extraction rate is 0.52%, and the purity is 83.12%.

[0093] Example 8

[0094] A1, 100g of camellia flowers is pulverized through a 100-mesh sieve and placed in a supercritical extraction device, and defatted and degelatinized at an extraction pressure of 20MPa, an extraction temperature of 55°C, and an extraction time of 60min; after the extraction is completed, the camellia flowers are taken out, and camellia flower powder A is obtained;

[0095] A2, the supercritically extracted camellia flower powder is pulverized, and 20 times of an ethanol aqueous solution is added to the supercritically extracted camellia flower powder, wherein the ethanol content is 60%; the ethanol aqueous solution of the sample is then controlled below 40°C, and then ultrasonicated for 60min;

[0096] A3, the solution in step A2 is controlled at 60°C, and stirred and extracted for 2.5h; after the extraction is completed, the temperature of the extract is lowered to room temperature, and filtration is performed to obtain a camellia extract, which is concentrated and dried to obtain a crude extract of camellia saponin A, the extraction rate is 5.61%, and the purity is 9.17%;

[0097] A4. The crude extract of camellia saponin A obtained in step A3 was added to 20 volumes of water according to the material-liquid ratio, and 1% α-amylase of the mass of dry material A was added to simulate the oral environment, and the reaction was carried out at 50°C and pH 7.0 for 2 hours; then 1% acid protease was added to simulate the gastric environment module, and the reaction was carried out at 50°C and pH 3.5 for 2 hours; then 1% trypsin and 1% lipase were added to simulate the intestinal environment module, and the reaction was carried out at 50°C and pH 7.5 for 2 hours; then the mixture was heated to 90°C, the enzyme was inactivated for 10 minutes, and the mixture was filtered to obtain a camellia saponin A solution.

[0098] A5. Purify the biomimetic extract obtained in step A4 using macroporous resin AB-8 with a sample load of 2 BV (BV is the bed volume of the resin), then elute with 2 BV of water and discard the eluate; then elute with 2 BV of 30% ethanol and discard the eluate, then elute with 70% extraction fraction anhydrous ethanol for 2 BV, and collect the eluate rich in camellia saponin A.

[0099] A6. The concentrated solution was concentrated to a solid content of 13%, then diluted with anhydrous ethanol to a solid content of 8%, allowed to stand at 8°C for 12 hours, and then filtered. Finally, the filtered concentrated solution was concentrated and dried to obtain camellia saponin A with an extraction yield of 0.56% and a purity of 73.12%.

[0100] Example 9

[0101] A1. 100 g of camellia flowers were crushed and passed through a 100-mesh sieve. The powder was placed in a supercritical extraction apparatus and subjected to degreasing and degelling at an extraction pressure of 30 MPa, an extraction temperature of 55°C, and an extraction time of 120 min. After the extraction, the camellia flowers were removed to obtain camellia powder A.

[0102] A2. Grind the supercritical camellia to obtain a powder; add 20 times the amount of ethanol aqueous solution, wherein the ethanol content is 60%, to the supercritical camellia powder; control the ethanol aqueous solution of the sample below 40° C. and then ultrasonicate for 60 minutes;

[0103] A3. The solution in step A2 was controlled at 60°C and stirred for extraction for 2 hours. After the extraction was completed, the extract was cooled to room temperature and filtered to obtain a camellia extract. The extract was concentrated and dried to obtain a crude extract of camellia saponin A with an extraction yield of 5.62% and a purity of 8.95%.

[0104] A4, the crude extract of camellia saponin A obtained in step A3 was added to water in a mass of 20 times the volume, 3% of the mass of dry substance A of α-amylase was added to simulate the oral environment, and reacted at 50°C and pH 7.0 for 2h; 3% of acid protease was then added to simulate the gastric environment module, and reacted at 50°C and pH 3.5 for 2h; 3% of trypsin and 3% of lipase were then added to simulate the intestinal environment module, and reacted at 50°C and pH 7.5 for 2h; then heated to 90°C to inactivate the enzyme for 10 minutes, and filtered to obtain a camellia saponin A solution.

[0105] A5, the biomimetic extract obtained in step A4 was purified by macroporous resin D-101, and the loading amount was 2BV (BV is the bed volume of the resin), then 2BV of water was used to elute and discard the eluate; then 2BV of 40% ethanol was used to elute and discard the eluate, and then 60% ethanol was used to elute 4BV, and the eluate rich in camellia saponin A was collected.

[0106] A6, the above concentrated solution was concentrated to a solid content of 10%, then diluted with anhydrous ethanol to a solid content of 8%, and then left still at 4°C for 12h, then filtered, and finally the filtered concentrated solution was concentrated and dried to obtain camellia saponin A, with an extraction rate of 0.54% and a purity of 81.25%.

[0107] Comparative Example 1

[0108] The preparation method of this comparative example is basically the same as that of Example 1, except that in step A1 of this comparative example, camellia flowers that have not been subjected to supercritical extraction are used as the raw material. In step A3, a crude extract of camellia saponin A is obtained, with an extraction rate of 0.8% and a purity of 8.5%; because this comparative example has not been subjected to supercritical extraction to remove fat and gel, the solution becomes a gel-like solid after concentration in step A6, and is still a gel-like solid after the addition of ethanol, as shown in FIG. 1, and cannot be subjected to subsequent crystallization and purification, so the gel-like solid after the addition of ethanol is directly dried to obtain camellia saponin A, and the camellia saponin A thus prepared has an extraction rate of 0.67% and a purity of 33.25%. Figure 1

[0109] Comparative Example 2

[0110] ​The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A4 of the present comparative example is different, and the specific operation is as follows: the crude extract of camellia saponin A obtained in step A3 is added into water with a mass of 50 times the volume, heated to 50°C, and pH is controlled at 7.0 for 2h; then the pH is controlled at 3.5 for 2h; then the pH is controlled at 7.5 for 2h; then heated to 90°C for 10 minutes, and filtered to obtain a camellia saponin A solution. Because the present comparative example does not go through the whole biomimetic process, the solution becomes a gel-like solid after concentration in step A6, and still is a gel-like solid after adding ethanol, as shown in Fig. 2, which cannot be crystallized and purified, so the gel-like solid after adding ethanol in the present comparative example is directly dried to obtain camellia saponin A. The extraction rate of camellia saponin A prepared by the present comparative example is 1.15%, and the purity is 34.16%. Figure 1

[0111] Comparative Example 3

[0112] The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A4 of the present comparative example is different, and the specific operation is as follows: 4% of α-amylase is added to simulate the oral environment, and the reaction is carried out at 50°C and pH 7.0 for 2h; no acidic protease is added to simulate the stomach environment module, and the reaction is carried out at 50°C and pH 3.5 for 2h; then 4% of trypsin and 4% of lipase are added to simulate the intestinal environment module, and the reaction is carried out at 50°C and pH 7.5 for 2h; then heated to 90°C for 10 minutes to inactivate the enzyme, and filtered to obtain a camellia saponin A solution. The extraction rate of camellia saponin A prepared by the present comparative example is 1.12%, and the purity is 39.12%.

[0113] Comparative Example 4

[0114] The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A4 of the present comparative example is different, and the specific operation is as follows: no α-amylase is added to simulate the oral environment, and the reaction is carried out at 50°C and pH 7.0 for 2h; 4% of acidic protease is added to simulate the stomach environment module, and the reaction is carried out at 50°C and pH 3.5 for 2h; then 4% of trypsin and 4% of lipase are added to simulate the intestinal environment module, and the reaction is carried out at 50°C and pH 7.5 for 2h; then heated to 90°C for 10 minutes to inactivate the enzyme, and filtered to obtain a camellia saponin A solution. The extraction rate of camellia saponin A prepared by the present comparative example is 1.05%, and the purity is 38.96%.

[0115] Comparative Example 5

[0116] ​The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A4 of the present comparative example is different, and the specific operation is as follows: 6% of the mass of dry matter A is added to simulate the oral environment of α-amylase, and the reaction is carried out at 50°C and pH 7.0 for 3h; 6% of the acid protease is added to simulate the stomach environment module, and the reaction is carried out at 50°C and pH 3.5 for 2h; no trypsin and lipase is added to simulate the intestinal environment module, and the reaction is carried out at 50°C and pH 7.5 for 3h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and then filtered to obtain a solution of camellia saponin A. The extraction rate of camellia saponin A prepared by this method is 1.08%, and the purity is 39.64%.

[0117] Comparative Example 6

[0118] The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A4 of the present comparative example is different, and the specific operation is as follows: 6% of the mass of dry matter A is added to simulate the oral environment of α-amylase, and the reaction is carried out at 50°C and pH 7.0 for 3h; 6% of the acid protease is added to simulate the stomach environment module, and the reaction is carried out at 50°C and pH 3.5 for 2h; no trypsin and lipase is added to simulate the intestinal environment module, and the reaction is carried out at 50°C and pH 7.5 for 3h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and then filtered to obtain a solution of camellia saponin A. The extraction rate of camellia saponin A prepared by this method is 1.08%, and the purity is 39.64%. Figure 1

[0119] Comparative Example 7

[0120] The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A4 of the present comparative example is different, and the specific operation is as follows: 6% of the mass of dry matter A is added to simulate the oral environment of α-amylase, and the reaction is carried out at 50°C and pH 7.0 for 3h; 6% of the acid protease is added to simulate the stomach environment module, and the reaction is carried out at 50°C and pH 3.5 for 2h; no trypsin and lipase is added to simulate the intestinal environment module, and the reaction is carried out at 50°C and pH 7.5 for 3h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and then filtered to obtain a solution of camellia saponin A. The extraction rate of camellia saponin A prepared by this method is 1.08%, and the purity is 39.64%. Figure 1

[0121] Comparative Example 8 ​​

[0122] The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A4 of the present comparative example is different, specifically: no a-amylase is added, and the reaction is carried out at 50°C and pH 7.0 for 3h; no acid protease is added, and the reaction is carried out at 50°C and pH 3.5 for 2h; 6% of trypsin and 6% of lipase are added to simulate the intestinal environment module, and the reaction is carried out at 50°C and pH 7.5 for 3h; then heated to 90°C, and the enzyme is inactivated for 10 minutes, and then filtered to obtain a camellia saponin A solution. Because the present comparative example does not undergo three-step enzymatic hydrolysis, the solution becomes a gel-like solid after concentration in step A6, and is still a gel-like solid after the addition of ethanol, as shown in FIG. 5, and cannot be subjected to subsequent crystallization and purification, so the gel-like solid after the addition of ethanol is directly dried to obtain camellia saponin A. The extraction rate of camellia saponin A prepared in this way is 0.74%, and the purity is 31.76%. Figure 1

[0123] Comparative Example 9

[0124] The preparation method of the present comparative example is basically the same as that of Example 1, except that the step A5 of the present comparative example is different, specifically:

[0125] In step A5, different concentrations of ethanol are not used for stepwise sample addition, but only 80% of the extraction fraction is eluted with anhydrous ethanol for 4BV, and the eluent rich in camellia saponin A is collected. Because the present comparative example does not undergo stepwise elution, the color of the subsequent sample is deep and the purity is low, and the extraction rate of camellia saponin A prepared in this way is 0.95%, and the purity is 40.76%.

[0126] Efficacy verification experiment

[0127] (1) Antioxidant capacity test: all the samples prepared in the above examples and comparative examples are subjected to antioxidant test of DPPH free radical scavenging rate, and the test method refers to the method published by Shanghai Daily Cosmetics Industry Association, i.e. Team Standard T / SHRH 006-2018, Cosmetics- Free Radical (DPPH) Scavenging Experimental Method. Shanghai Daily Cosmetics Industry Association, 2018. The concentration of free radical scavenger when the free radical scavenging rate is 50% is IC50 value, and IC50 value is a commonly used index for evaluating the effect of free radical scavenger. The smaller the value, the smaller the concentration of free radical scavenger required to achieve 50% free radical scavenging rate, and the better the free radical scavenging effect.

[0128] Table 2: Antioxidant test results of examples and comparative examples

[0129]

[0130]

[0131] ​(ii) Anti-glycation test: Advanced glycation end products have two sources: 1. Glycation reaction accompanied by oxidation; 2. Lipid peroxidation products and protein reaction. Advanced glycation end products are numerous, most of which have fluorescence, with an absorption peak at 370 nm and an emission peak at 445 nm. Using this feature, advanced glycation end products can be tested by fluorescence method. The test method refers to the literature Anti-Glycation Activities of Phenolic Constituents from Silybum marianum (Milk Thistle) Flower in Vitro and on Human Explants; Isoferulic acid, a new anti-glycation agent, inhibits fructose- and glucose-mediated protein glycation in vitro (Meeprom, Sompong et al. 2013)

[0132] Table 3 Example and comparative example anti-glycation test results

[0133]

[0134] (iii) Repair efficacy test: Epidermal keratinocytes are important cells that make up the epidermal layer. When the skin surface is damaged, keratinocytes will be stimulated to migrate and repair the damaged part. When the in vitro cells grow to a single layer state, an empty area is artificially created on the monolayer cells, and the cells at the edge of the scratch will gradually enter the empty area to heal the scratch, to some extent simulating the in vivo cell migration process. By measuring the cell migration rate after sample treatment, it is evaluated whether the test substance has repair efficacy. The efficacy of the composition is verified by testing the migration rate of epidermal keratinocytes of all examples and comparative examples described in the composition.

[0135] Table 4 Example and comparative example cell migration rate test results

[0136] Sample Added Concentration (mg / mL) Cell Migration Rate (%) Example 1 1.0 35.49％ Example 2 1.0 31.19％ Example 3 1.0 46.49％ Example 4 1.0 26.57％ Example 5 1.0 32.96％ Example 6 1.0 31.26％ Example 7 1.0 65.35％ Example 8 1.0 58.94％ Example 9 1.0 63.59％ Comparative Example 1 1.0 9.64％ Comparative Example 2 1.0 13.48％ Comparative Example 3 1.0 17.25％ Comparative Example 4 1.0 16.98％ Comparative Example 5 1.0 19.23％ Comparative Example 6 1.0 6.59％ Comparative Example 7 1.0 8.49％ Comparative Example 8 1.0 7.64％ Comparative Example 9 1.0 23.49％

[0137] (IV) Whitening efficacy test: Melanin is a high-molecular biological pigment, essentially a protein, usually in a polymerized form in animal skin or hair, even in plants and protists, its content and distribution will directly affect the color of the skin, hair and eyes. Melanin is mainly composed of two quinone polymers, eumelanin and pheomelanin, and its biosynthesis is a series of biochemical reaction processes initiated by the hydroxylation of tyrosine catalyzed by tyrosinase. Tyrosinase is the rate-limiting enzyme of melanin production, and its quantity and activity determine the speed and yield of melanin production. B16-F10 melanocyte cell line is a typical cell line for studying cell melanin production. In this experiment, the relative content of melanin production was determined to evaluate the melanin inhibition ability of the test sample, i.e. whitening efficacy. The test method refers to Kim J H, Baek S H, Kim D H, et al. Downregulation of Melanin Synthesis by Haginin A and Its Application to In Vivo Lightening Model [J]. Journal of Investigative Dermatology, 2008, 128(5): 1227-35.

[0138] Table 5 Test results of cell melanin inhibition rate of examples and comparative examples

[0139]

[0140] (VI) Anti-wrinkle efficacy test: all the above example and comparative example compositions are verified by promoting fibroblast type I, III and V collagen synthesis experiment to verify the anti-wrinkle effect of the composition, the experimental method refers to T / SHRH 031-2020, Cosmetics tightening, anti-wrinkle efficacy test - in vitro fibroblast type I collagen content determination. Shanghai daily use cosmetics industry association, 2018; Cansn Güngrmü, Dürdane Kolankaya. Characterization of type I, III and V collagens in high-density cultured tenocytes by triple-immunofluorescence technique [J]. Cytotechnology, 2008, 58(3): 145-152.

[0141] Table 6 Test results of type I, III and V collagen promotion rate of examples and comparative examples

[0142]

[0143] Application Example 1

[0144] The application example provides application and preparation method of camellia saponin A in skin care cream formula, and the specific formula is as follows:

[0145] Phase A

[0146]

[0147]

[0148] Phase B

[0149]

[0150] Phase C

[0151] SIMULGEL NS 0.8%;

[0152] Phase D

[0153] Camellia saponin A 5.0%;

[0154] Fragrance 0.1%;

[0155] The preparation method is as follows:

[0156] A, under normal temperature, the A item is added into the water pot in turn, fully stirred and dispersed uniformly, then heated to 90±2℃, stirred uniformly, and then pumped into the emulsifying pot and homogenized for 5 minutes;

[0157] B, under normal temperature, the B item is added into the oil pot in turn, then heated to 85±2℃, completely dissolved, and then pumped into the emulsifying pot;

[0158] C, homogenized for 10 minutes, the C item is added, and kept for 15 minutes, and stirred uniformly;

[0159] D, 50±2℃, the D item is added, and stirred uniformly.

[0160] Application Example 2

[0161] The application example provides application and preparation method of camellia saponin A in emulsion formula, and the specific formula is as follows:

[0162] Phase A

[0163]

[0164]

[0165] Phase B

[0166] Caprylic / capric triglyceride 5%;

[0167] MONTANOV 68MB 0.5%;

[0168] C phase

[0169] Trometamol 0.04%

[0170] D phase

[0171] SIMULGEL NS 0.3%;

[0172] E phase

[0173] Camellia saponin A 5.0%;

[0174] Fragrance 0.06%;

[0175] The preparation method is as follows:

[0176] A. At room temperature, the A item is added into a water pot in turn, fully stirred and dispersed uniformly, and then heated to 90±2℃, stirred uniformly, and then pumped into an emulsifying pot and homogenized for 5 minutes.

[0177] B. At room temperature, the B item is added into an oil pot in turn, and then heated to 85±2℃, completely dissolved, and then pumped into an emulsifying pot.

[0178] C. Homogenized for 10 minutes, the C item is added, and kept at 15 minutes, and then stirred uniformly.

[0179] D. At 50±2℃, the D item is added, and then stirred uniformly.

[0180] Application Example 3

[0181] The application example provides an application and a preparation method of camellia saponin A in a water agent formula, and the specific formula is as follows:

[0182] A phase

[0183]

[0184]

[0185] B phase

[0186] Trometamol 0.01%

[0187] C phase

[0188] Camellia saponin A 5.0%;

[0189] D phase

[0190] Fragrance solubilizer 0.05%

[0191] Fragrance 0.01%;

[0192] A, at room temperature, the A item is added into the water pot in turn, fully stirred and dispersed uniformly, then heated to 90±2℃, stirred uniformly, and then pumped into the emulsifying pot and homogenized for 5 minutes;

[0193] B, at room temperature, the B item is added into the oil pot in turn, then heated to 85±2℃, fully dissolved, and then pumped into the emulsifying pot.

[0194] C, homogenized for 10 minutes, the C item is added, and kept for 15 minutes, and then stirred uniformly;

[0195] D, at 50±2℃, the D item is added, and then stirred uniformly.

[0196] To sum up, the application provides a method for extracting anti-aging component sasanqua saponin A from sasanqua based on full bionic targeting technology and application, and particularly relates to a method for realizing efficient enrichment and structure protection of sasanqua saponin A by simulating the whole process of human oral cavity-gastric-intestinal decomposition and absorption. The saponin yield is ≤1.0% by using the traditional alcohol extraction method (CN 107280993A); and the microwave-assisted method can cause degradation of heat-sensitive components. The existing technology of biological enzymatic hydrolysis method mainly uses single cellulase treatment, which cannot effectively break the complex structure (containing lignin-pectin covalent cross-linking network) of sasanqua flower or leaf cell wall, and the enzymatic hydrolysis efficiency is low. The application uses supercritical extraction technology to pretreat sasanqua flowers or leaves, removes lipids and gel impurities which are easy to form gelatinous substances, and provides convenience for subsequent purification; and applies full bionic enzymatic hydrolysis technology to solve the problems of low sasanqua saponin A yield and easy degradation, and the full bionic extraction can degrade lipids, proteins, tannins, sugars and other impurities, thereby providing convenience for subsequent crystallization; then macroporous resin adsorption technology is used to purify sasanqua saponin A to obtain a purified sasanqua saponin A solution; and finally, sasanqua saponin A is obtained through crystallization purification. The sasanqua saponin A can achieve the effects of anti-wrinkle, repair and whitening through multiple pathways and multiple directions, and can better achieve the effects of improving wrinkles, repairing skin, brightening skin color and delaying aging, and can be widely applied to various types of cosmetics. The application realizes the enrichment of sasanqua saponin A in the extract by conventional extraction and subsequent full bionic extraction simulating the oral cavity-gastric-intestinal tract, so that the content of sasanqua saponin A in the final extract is significantly improved, and the purity is more than 60%.

[0197] The specific embodiments of the application are described above. It should be understood that the application is not limited to the above specific embodiments, and those skilled in the art can make various changes or modifications within the scope of the claims, which do not affect the essential content of the application. The embodiments of the present application and the features in the embodiments can be combined with each other arbitrarily without conflict.

Claims

1. A method for extracting camellia saponin A from camellia, characterized in that: The method comprises the following steps: A1. Take camellia flowers or leaf powder and perform supercritical extraction; The powders obtained by extraction of A2 and A1 were ultrasonically treated with anhydrous ethanol, stirred and extracted, filtered to obtain a camellia extract, and concentrated and dried to obtain a crude extract of camellia saponin A; A3, adding the crude extract of camellia saponin A to water, adding α-amylase to simulate the oral environment, and reacting at 30-50° C. with the pH range controlled between 6.0-7.0 for 0.5-2 h; then adding an acidic protease module to simulate the gastric environment, and reacting at 40-50° C. with the pH range controlled between 1.5-3.5 for 0.5-2 h; then adding trypsin and lipase to simulate the intestinal environment, and reacting at 40-50° C. with the pH range controlled between 6.5-7.5 for 0.5-2 h; then heating to inactivate the enzymes, and filtering to obtain a biomimetic extract of camellia saponin A; A4, purifying the camellia saponin A biomimetic extract with a macroporous resin, and collecting the eluate rich in camellia saponin A; A5. Concentrate the camellia saponin A eluate to a solid content of 10-16%, then dilute it with anhydrous ethanol to a solid content of 5-8%, let it stand at 2-8° C. for 10-15 hours, and then filter it; concentrate and dry the filtrate to obtain camellia saponin A.

2. The method for extracting camellia saponin A from camellia according to claim 1, characterized in that In step A1, the supercritical extraction has an extraction pressure of 15-40 MPa, an extraction temperature of 45-55° C., and an extraction time of 30 min-120 min.

3. The method for extracting camellia saponin A from camellia according to claim 1, wherein In step A1, the camellia flower or leaf powder is prepared by crushing the camellia flower or leaf through a 100-160 mesh sieve.

4. The method for extracting camellia saponin A from camellia according to claim 1, wherein In step A2, the powder obtained by extraction in step A1 is added to 30-80% volume fraction anhydrous ethanol at a mass ratio of 10-30 times that of the solid-liquid ratio, the solution temperature is controlled below 40° C., and ultrasonic treatment is performed.

5. The method for extracting camellia saponin A from camellia according to claim 1 or 4, characterized in that The ultrasonic power of the ultrasonic treatment is 500W-3000W, and the ultrasonic time is 10min-60min.

6. The method for extracting camellia saponin A from camellia according to claim 1, characterized in that In step A2, the temperature of the solution during the stirring extraction is controlled at 40° C.-70° C., and the stirring extraction time is 1 h-4 h.

7. The method for extracting camellia saponin A from camellia according to claim 1, characterized in that Step A3 includes at least one of the following technical features: B1. Add the crude extract of camellia saponin A to water in a volume of 10-50 times its mass according to the solid-liquid ratio; B2, the addition amount of the α-amylase is 0.5-3% of the mass of the crude extract of camellia saponin A; B3, the amount of the acidic protease added is 0.5-3% of the crude extract of camellia saponin A; B4, the amount of trypsin added is 0.5-3% of the mass of the crude extract of camellia saponin A; B5, the amount of lipase added is 0.5-3% of the crude extract of camellia saponin A; B6. Heat the enzyme to inactivate it at 80-100°C for 5-20 minutes.

8. The method for extracting camellia saponin A from camellia according to claim 1, characterized in that In step A4, The macroporous resin is one of YKDH-5, D-101, AB-8, X-5, and NKA-9; And / or, in the macroporous resin purification, the loading amount of the camellia saponin A biomimetic extract is 1-3BV, where BV is the bed volume of the resin; then eluted with 1-3BV of water and the eluate is discarded; then eluted in sections with 30-80% extraction fraction anhydrous ethanol for 1-6BV, and the eluate rich in camellia saponin A is collected.

9. A camellia saponin A extract obtained by the method according to any one of claims 1 to 8.

10. Use of the camellia saponin A extract according to claim 9 in cosmetics.

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