Camellia nitidissima leaf saponin extract as well as extraction method and application thereof

Abstract

The invention discloses a camellia nitidissima leaf saponin extract as well as an extraction method and application thereof, and relates to the technical field of natural products. The invention develops application of the camellia nitidissima leaf saponin in the field of cosmetics, further in the field of whitening and in promoting absorption of effective components, and tests show that the extraction purity of the camellia nitidissima leaf saponin extracted by the extraction method of the camellia nitidissima leaf saponin extract provided by the invention can be as high as 83%; a melanoma cell activity inhibition test is carried out on an aqueous extract obtained by water extraction through further comparison, when the dosage is equal, the melanoma cell activity inhibition rate of the saponin extract of the camellia nitidissima leaves is higher than that of the aqueous extract, and in addition, the application further finds that the saponin of the camellia nitidissima leaves has the effect of promoting absorption and permeation of functional components, and has the effect of inhibiting melanoma cell activity. The camellia nitidissima saponin also has a relatively remarkable effect on polypeptide absorption, and test results prove that the skin transmittance of the polypeptide can be remarkably increased by adding the camellia nitidissima saponin, so that the camellia nitidissima saponin has a good application prospect in the field of cosmetics.

Description

Technical Field

[0001] The present application relates to the technical field of natural products, and in particular to a golden camellia saponin extract, an extraction method and applications thereof. Background Art

[0002] Yellow Camellia is an evergreen shrub or small tree of the Camellia genus of the Theaceae family, growing in limestone mountains at an altitude of 130 to 460 meters. It is subdivided into Chrysanthae Chang and Camellia aurea Chang according to the number of styles. Yellow Camellia was first discovered in 1924. Unlike the red or white petals of other Camellia plants, the petals of Yellow Camellia are waxy golden yellow, hence the name.

[0003] According to previous reports, golden camellia tea is used in various important compositions, such as the "Double Golden Camellia Chinese Medicine Composition and Its Preparation Method and Application" disclosed in the Chinese patent "CN115025163B", which uses golden camellia tea as one of the main effective components, and the prepared composition is used to prepare drugs for treating diabetic kidney damage and for preparing drugs for inhibiting kidney swelling and pain; it can be seen that golden camellia tea has a high development value in the field of medical care. At present, there are few reports on the application of saponin extracts of golden camellia tea, especially in the fields of cosmetics and whitening. This application aims to further study golden camellia tea extracts in this field to fill the gap in this field. Summary of the invention

[0004] In view of this, the purpose of the present application is to provide a camellia chrysantha saponin extract, an extraction method and application thereof.

[0005] In a first aspect, the present application provides a method for extracting a golden camellia saponin extract, the extraction method comprising the following steps:

[0006] S1. Preheat the ultrasonic cleaning machine to 45-55°C, put the golden camellia tea powder into a conical flask, and add ethanol;

[0007] S2. Add an appropriate amount of 0.1 mol / L dilute hydrochloric acid to adjust the pH to 4, then add cellulase and shake well;

[0008] S3, place the conical flask in an ultrasonic cleaning machine, extract under ultrasound for 45-55 minutes, take the macroporous resin wet method column, and load the sample;

[0009] S4. After adsorption for 24 hours, the mixture was eluted with 75% ethanol, the effluent was collected, and evaporated to dryness in a water bath to obtain the golden camellia saponin extract.

[0010] Preferably, in step S1, the mass of golden camellia tea is 1 g;

[0011] In step S2, the volume fraction of ethanol is 55%-58%; 4%-4.5% of cellulase is added;

[0012] In step S3, the ultrasonic power is 400 W; the macroporous resin is 10 g of D101 macroporous resin, and the flow rate of the extract during loading is 0.5 mL min -1 , volume 1BV;

[0013] In step S4, 1BV of liquid is discharged.

[0014] In a second aspect, the present application provides a delivery system comprising a golden camellia saponin extract, wherein the golden camellia saponin extract is extracted by any of the extraction methods described above;

[0015] The delivery systems include liposomes, cyclodextrins, Pickering emulsions, and hydrogels.

[0016] In a third aspect, the present application provides a method for preparing golden camellia saponin liposomes, the preparation method comprising the following steps:

[0017] Weigh golden camellia tea saponin, add PBS buffer with a pH of 7.5-8.5, and ultrasonically dissolve for 4-6 minutes before use; the golden camellia tea saponin is extracted by any of the extraction methods described above;

[0018] Weigh lecithin and cholesterol, dissolve them in methanol in a round-bottom flask, and evaporate them at 40-50°C until a uniform film forms;

[0019] The mass ratio of the golden camellia saponin, the lecithin and the cholesterol is 2:4:1.

[0020] In a fourth aspect, the present application provides a method for preparing golden camellia saponin cyclodextrin, the preparation method comprising the following steps:

[0021] Weighing cyclodextrin and golden camellia saponin, placing the cyclodextrin in a culture dish, and adding water to fully dissolve it; the golden camellia saponin is extracted by any of the extraction methods described above;

[0022] Dissolve golden camellia saponin in water, then slowly drop the golden camellia saponin solution into the cyclodextrin solution, stir, centrifuge the obtained solution at 10°C and 14000r / min for 1h, remove the supernatant, and then freeze in a freeze dryer until completely dry to obtain a white solid powder;

[0023] The mass ratio of the cyclodextrin to the golden camellia tea is 2:1, the inclusion time is 2.5 hours, and the inclusion temperature is 4°C.

[0024] In a fifth aspect, the present application provides a golden camellia saponin pearl cream, comprising the following components:

[0025] Phase A: Camellia seed oil 3%, isononyl isononanoate 5%, caprylic / capric triglyceride 3%, bisabolol 0.2%, tocopheryl acetate 0.5%, lemon essential oil 0.1%, emulsifier A165 1.89%, emulsifier M82 1.11%;

[0026] Item B: 3.5% glycerol, 2% propylene glycol, 1.5% trehalose, 2% 1-3 propanediol, 0.35% carbomer U10, 0.1% β-glucan, 0.1% glabridin, 0.02% EDTA-2Na, add water to 100%;

[0027] Phase C: 0.5% PEG-40 hydrogenated castor oil, 4% pearl hydrolyzate, 0.5% phenoxyethanol, 0.05% golden camellia saponin; the golden camellia saponin is extracted by any of the extraction methods described above;

[0028] Preferably, the golden camellia tea saponin pearl cream is prepared by the following method:

[0029] The components of the phase A, the phase B, and the phase C are mixed and placed in different beakers to be evenly dispersed for later use;

[0030] Take 2g of sodium hydroxide solid in a conical flask, add 18g of ultrapure water to make a 10% sodium hydroxide solution, which is used as the acid-base regulator in this experiment;

[0031] Place the beaker of phase C in a constant temperature water bath at 55-65℃, stir and dissolve until uniform, and set aside;

[0032] Add phase A into the beaker of phase B, turn on the homogenizer, and homogenize for 5 to 10 minutes at a speed of 3000 r / min;

[0033] Add acid-base regulator to adjust pH value and continue homogenizing and emulsifying for 2-4 minutes;

[0034] When the cream is cooled to 40-60℃ and kept for more than 30 minutes, add the evenly mixed phase C to the cream, homogenize and emulsify at 3000r / min for 3-5 minutes, stir evenly and keep warm for 20 minutes to obtain the base cream.

[0035] In the sixth aspect, the present application provides a kind of golden camellia saponin extracted by the extraction method of any of the golden camellia saponin extracts described above, or golden camellia saponin liposomes prepared by the preparation method of golden camellia saponin liposomes described above, or golden camellia saponin cyclodextrin prepared by the preparation method of flower tea saponin cyclodextrin described above, or the use of golden camellia saponin pearl cream described above in promoting the absorption and penetration of effective ingredients.

[0036] In the seventh aspect, the present application provides a golden camellia saponin extracted by the extraction method of any of the above-mentioned golden camellia saponin extracts, or golden camellia saponin liposomes prepared by the method for preparing golden camellia saponin liposomes described above, or golden camellia saponin cyclodextrin prepared by the method for preparing flower tea saponin cyclodextrin described above, or the use of golden camellia saponin pearl cream based on any of the above-mentioned methods in inhibiting tyrosinase activity.

[0037] In the eighth aspect, the present application provides a golden camellia saponin extracted by the extraction method of any of the above-mentioned golden camellia saponin extracts, or golden camellia saponin liposomes prepared by the method for preparing golden camellia saponin liposomes described above, or golden camellia saponin cyclodextrin prepared by the method for preparing flower tea saponin cyclodextrin described above, or golden camellia saponin pearl cream based on any of the above-mentioned for use in repairing photoaging damage caused by ultraviolet radiation.

[0038] Compared with the prior art, the beneficial effects of the present application are as follows: the present application develops the application of golden camellia saponins in the field of cosmetics, and further applies them to the whitening field; among them, it has been tested that the extraction purity of golden camellia saponins extracted by the extraction method of golden camellia saponin extract provided by the present application can be as high as 83%; by further comparing the water extract obtained by water extraction for melanoma cell activity inhibition test, under the condition of equal dose, the inhibition rate of golden camellia saponin extract on melanoma cell activity is significantly higher than that of water extract; in addition, the present application further finds that golden camellia saponins have a more significant effect on promoting the absorption and penetration of effective ingredients, such as polypeptide absorption. The test results show that the addition of golden camellia saponins can significantly increase the skin permeability of polypeptides, so it has good application prospects in the field of cosmetics. The present application fills the gap in golden camellia related extracts in this field, and provides a good research basis and reference for subsequent in-depth research. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0040] Figure 1 This is a graph showing the test results of the effect of golden camellia saponin extract on the catalytic activity of tyrosine monophenolase in one embodiment of the present application;

[0041] Figure 2 This is a graph showing the test results of the effect of golden camellia saponin extract on the catalytic activity of tyrosine polyphenolase in one embodiment of the present application;

[0042] Figure 3 This is a graph showing the test results of the effect of golden camellia saponin extract on the activities of IL-1, IL-6, MDA and SOD in one embodiment of the present application;

[0043] Figure 4 This is a graph showing the test results of the effect of golden camellia saponin extract on A375 melanoma cells in one embodiment of the present application;

[0044] Figure 5 This is a graph showing the test results of the effect of the golden camellia saponin extract in promoting the absorption and penetration of polypeptides in one embodiment of the present application. DETAILED DESCRIPTION

[0045] In order to make the purpose, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with embodiments. Several embodiments of the present invention are given below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present invention more thorough and comprehensive:

[0046] The present application provides a method for extracting a golden camellia saponin extract, the extraction method comprising the following steps:

[0047] S1. Preheat the ultrasonic cleaning machine to 45-55°C, put the golden camellia powder in a conical flask, and add ethanol; it is understandable that preheating the ultrasonic machine first can provide favorable preconditions for the extraction of the golden camellia target extract, allowing it to be kept at a suitable temperature during the ultrasonic reaction, which is helpful to improve the extraction rate and extraction purity of the target extract.

[0048] S2. Add an appropriate amount of 0.1 mol / L dilute hydrochloric acid to adjust the pH to 4, then add cellulase and shake well; it can be understood that in this scheme, dilute hydrochloric acid is added and the pH is adjusted to 4, so that the golden camellia tea is first placed in an acidic condition, which is conducive to destroying the cell structure of the golden camellia tea, thereby allowing the cell contents to flow out, thereby improving the extraction rate and extraction purity of the target product; this scheme further adds cellulase and shakes well. The addition of cellulase can further accelerate the destruction of the plant cell wall, which not only effectively improves the extraction efficiency, but also assists in improving the extraction rate and extraction purity.

[0049] S3. Place the conical flask in an ultrasonic cleaning machine, extract under ultrasound for 45-55 minutes, take a macroporous resin wet column, and load the sample; it can be understood that this scheme first uses ultrasound for extraction, combined with the acid treatment and cellulase treatment of golden camellia tea in the pre-reaction, which can further improve the outflow of the target extract.

[0050] S4. After adsorption for 24 hours, the mixture was eluted with 75% ethanol, the effluent was collected, and evaporated to dryness in a water bath to obtain the golden camellia saponin extract.

[0051] Further, in one embodiment, in step S2, the volume fraction of ethanol is 55%-58%; 4%-4.5% of cellulase is added;

[0052] In step S3, the ultrasonic power is 400 W; the macroporous resin is 10 g of D101 macroporous resin, and the flow rate of the extract during loading is 0.5 mL min -1 , volume 1BV;

[0053] In step S4, 1BV of liquid is discharged.

[0054] The present application also provides a delivery system comprising a golden camellia saponin extract, wherein the golden camellia saponin extract is extracted by the extraction method described above.

[0055] Furthermore, the delivery system includes but is not limited to liposomes, cyclodextrins, Pickering emulsions, and hydrogels.

[0056] The present application also provides a method for preparing golden camellia saponin liposomes, the preparation method comprising the following steps: weighing golden camellia saponin, adding PBS buffer with a pH of 7.5-8.5, ultrasonically dissolving for 4-6 minutes and then setting aside; wherein the golden camellia saponin is extracted by the extraction method described above;

[0057] Weigh lecithin and cholesterol, dissolve them in methanol in a round-bottom flask, and evaporate them at 40-50°C until a uniform film forms;

[0058] The mass ratio of the golden camellia saponin, the lecithin and the cholesterol is 2:4:1.

[0059] The present application also provides a method for preparing golden camellia saponin cyclodextrin, the preparation method comprising the following steps:

[0060] Weighing cyclodextrin and golden camellia saponin, placing cyclodextrin in a culture dish, and adding water to fully dissolve it; the golden camellia saponin is extracted by the above-mentioned extraction method;

[0061] Dissolve golden camellia saponin in water, then slowly drop the golden camellia saponin solution into the cyclodextrin solution, stir, centrifuge the obtained solution at 10°C and 14000r / min for 1h, remove the supernatant, and then freeze in a freeze dryer until completely dry to obtain a white solid powder;

[0062] Furthermore, in one embodiment, the mass ratio of cyclodextrin to golden camellia tea is 2:1, the inclusion time is 2.5h, and the inclusion temperature is 4°C.

[0063] The present application also provides a golden camellia saponin pearl cream, comprising the following components:

[0064] Phase A: Camellia seed oil 3%, isononyl isononanoate 5%, caprylic / capric triglyceride 3%, bisabolol 0.2%, tocopheryl acetate 0.5%, lemon essential oil 0.1%, emulsifier A165 1.89%, emulsifier M82 1.11%;

[0065] Item B: 3.5% glycerol, 2% propylene glycol, 1.5% trehalose, 2% 1-3 propanediol, 0.35% carbomer U10, 0.1% β-glucan, 0.1% glabridin, 0.02% EDTA-2Na, add water to 100%;

[0066] Phase C: 0.5% PEG-40 hydrogenated castor oil, 4% pearl hydrolyzate, 0.5% phenoxyethanol, 0.05% golden camellia saponin; the golden camellia saponin is extracted by the above extraction method.

[0067] Furthermore, the golden flower tea saponin pearl cream is prepared by the following method:

[0068] Mix the above-mentioned components of phase A, phase B and phase C respectively, place them in different beakers, mix and disperse them evenly for later use;

[0069] Place the oil phase beaker and the water phase beaker in a constant temperature water bath at 75-85℃, and use a glass cup to stir and dissolve in the beaker until uniform, and set aside;

[0070] Take 2g of sodium hydroxide solid in a conical flask, add 18g of ultrapure water to make a 10% sodium hydroxide solution, which is used as the acid-base regulator in this experiment;

[0071] Place the beaker of phase C in a constant temperature water bath at 55-65℃, stir and dissolve until uniform, and set aside;

[0072] Add phase A into the beaker of phase B, turn on the homogenizer, and homogenize for 5 to 10 minutes at a speed of 3000 r / min;

[0073] Add acid-base regulator to adjust pH value and continue homogenizing and emulsifying for 2-4 minutes;

[0074] When the cream is cooled to 40-60℃ and kept for more than 30 minutes, add the evenly mixed phase C to the cream, homogenize and emulsify at 3000r / min for 3-5 minutes, stir evenly and keep warm for 20 minutes to obtain the base cream.

[0075] The present application also provides a golden camellia saponin extracted by the extraction method of any of the golden camellia saponin extracts described above, or a golden camellia saponin liposome prepared by the method for preparing golden camellia saponin liposomes described above, or a golden camellia saponin cyclodextrin prepared by the method for preparing flower tea saponin cyclodextrin described above, or a golden camellia saponin pearl cream based on any of the above for use in promoting the absorption and penetration of effective ingredients.

[0076] The present application also provides a golden camellia saponin extracted by the extraction method of any of the above-mentioned golden camellia saponin extracts, or a golden camellia saponin liposome prepared by the above-mentioned method for preparing golden camellia saponin liposomes, or a golden camellia saponin cyclodextrin prepared by the above-mentioned method for preparing flower tea saponin cyclodextrin, or a golden camellia saponin pearl cream based on any of the above-mentioned methods for inhibiting tyrosinase activity.

[0077] The present application also provides a golden camellia saponin extracted by the extraction method of any of the above-mentioned golden camellia saponin extracts, or golden camellia saponin liposomes prepared by the above-mentioned method for preparing golden camellia saponin liposomes, or golden camellia saponin cyclodextrin prepared by the above-mentioned method for preparing flower tea saponin cyclodextrin, or a golden camellia saponin pearl cream based on any of the above-mentioned methods for repairing photoaging damage caused by ultraviolet radiation.

[0078] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0079] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0080] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0081] Example 1

[0082] This embodiment provides a method for extracting a golden camellia saponin extract, the extraction method comprising the following steps:

[0083] S1. Preheat the ultrasonic cleaning machine to 50°C, weigh 1g of golden camellia tea powder and put it into a conical flask, add 50mL of 56.12% volume fraction ethanol and shake well;

[0084] S2. After adding an appropriate amount of 0.1 mol / L dilute hydrochloric acid to adjust the pH to 4, add 4.09% cellulase and shake well;

[0085] S3, placing the conical flask in an ultrasonic cleaning machine, extracting at an ultrasonic power of 400 W for 49.02 min, and loading 10 g of D101 macroporous resin into a column by wet method;

[0086] S4, the extraction solution flows at a rate of 0.5 mL min -1 The sample was loaded at a volume of 1 BV. After adsorption for 24 hours, it was eluted with 75% ethanol. The effluent was 1 BV. The effluent was collected and evaporated to dryness in a water bath.

[0087] The total saponin yield in this example is 5.1% and the purity is 83%.

[0088] Comparative Example 1

[0089] This comparative example provides a method for extracting golden camellia saponin extract.

[0090] Accurately weigh 1.g of golden camellia tea powder and put it in a conical flask, add 80% ethanol at a solid-liquid ratio of 1:30 (g / ml), extract at 35°C for 30 minutes. Take 10g of AB macroporous resin and wet-pack it into a column. Load the extract at a flow rate of 0.5mL·min-1 and a volume of 1BV. After adsorption for 24 hours, add 50% ethanol, 1BV of the effluent, collect the effluent, and evaporate it in a water bath.

[0091] The total saponin yield in this comparative example is 3.2% and the purity is 69%.

[0092] Example 2

[0093] This embodiment provides a method for preparing golden camellia saponin liposomes, which comprises the following steps:

[0094] Weigh 0.1g of golden camellia saponin, add 15ml of PBS buffer with a pH of 8.0, sonicate for 5min and set aside. Weigh 0.2g of lecithin and 0.05g of cholesterol, dissolve them in 20ml of methanol in a 250ml round-bottom flask, and evaporate them at 45°C until a uniform film is formed.

[0095] The encapsulation efficiency in this example is 66.14%.

[0096] Example 3

[0097] This embodiment provides a method for preparing golden camellia saponin cyclodextrin, which comprises the following steps:

[0098] Weigh cyclodextrin and golden camellia saponin, place the former in a culture dish, add water to fully dissolve it; slowly drop the latter into the former solution, stir for a certain time at a certain temperature, centrifuge the obtained solution at 10°C and 14000r / min for 1h, remove the supernatant, place in a freeze dryer and freeze until completely dry to obtain the product (white solid powder).

[0099] Among them, the feed ratio is 2:1, the inclusion time is 2.5h, and the inclusion temperature is 4℃.

[0100] The inclusion rate in this embodiment can reach 55.06%.

[0101] Example 4

[0102] This embodiment provides a golden camellia tea saponin pearl cream and a preparation method thereof, wherein the golden camellia tea saponin pearl cream comprises the following components:

[0103] Phase A: Camellia seed oil 3%, isononyl isononanoate 5%, caprylic / capric triglyceride 3%, bisabolol 0.2%, tocopheryl acetate 0.5%, lemon essential oil 0.1%, emulsifier A165 1.89%, emulsifier M82 1.11%;

[0104] Item B: glycerol 3.5%, propylene glycol 2%, trehalose 1.5%, 1-3 propanediol 2%, carbomer U10 0.35%, β-glucan 0.1%, glabridin 0.1%, EDTA-2Na 0.02%, water added to 100%;

[0105] Phase C: PEG-40 hydrogenated castor oil 0.5%, pearl hydrolyzate 4%, phenoxyethanol 0.5%, golden camellia saponin 0.05%.

[0106] The preparation method specifically comprises the following steps:

[0107] Mix the components of phase A, phase B and phase C separately, place them in different beakers, mix and disperse them evenly for later use.

[0108] Take 2g of sodium hydroxide solid in a conical flask, add 18g of ultrapure water to make a 10% sodium hydroxide solution, which is used as the acid-base regulator in this experiment.

[0109] Place the beaker of phase C in a constant temperature water bath of 55-65℃, stir and dissolve until uniform, and set aside.

[0110] Add phase A into the beaker of phase B, turn on the homogenizer, and homogenize for 5 to 10 minutes at a speed of 3000 r / min.

[0111] Add acid-base regulator to adjust pH value, and continue homogenizing and emulsifying for 2-4 minutes. When the cream cools to 40-60℃, keep it for more than 30 minutes.

[0112] Then add the evenly mixed phase C into the cream, homogenize and emulsify at 3000r / min for 3-5min, stir evenly and keep warm for 20min to obtain the base cream.

[0113] Example 5

[0114] This study examined the inhibitory activity of tyrosinase and monophenolase on the saponin extract of Camellia chrysantha

[0115] Using L-tyrosine as substrate, the inhibitory effect of golden camellia saponins on the monophenolase activity of tyrosinase was determined. 0.3 mL of phosphate buffer and 2.0 mL of 1 mmol / L tyrosine solution were added to the test tubes, mixed thoroughly, and incubated at 25°C for 10 min. Then, 0.3, 0.6, 0.9, 1.2, and 1.5 mg / mL of golden camellia saponins solution were added, and 0.1 mL of 200 U / mL tyrosinase solution was added. The wavelength of the microplate reader was 475 nm, and the absorbance change within 5 to 10 min was measured at 1 min intervals.

[0116] For detailed results, see Figure 1 Among them, the activity of tyrosine monophenolase can be significantly inhibited by 0.3-1.5 mg / mL saponin. As the mass concentration of saponin increased from 0.9 mg / mL to 1.2 mg / mL, the inhibition rate increased from 45.42% to 51.05%, and the IC50 value was 1.067 mg / mL.

[0117] Example 6

[0118] This study examined the inhibitory effect of golden camellia saponin extract on tyrosinase and polyphenolase activity.

[0119] Using L-DOPA as substrate, the inhibitory effect of golden camellia tea saponins on the polyphenolase activity of tyrosinase was determined. 0.8 mL of phosphate buffer and 1.5 mL of L-DOPA were added to the test tube, incubated at 25°C for 10 min, 0.1 mL of 0.3-3.0 mg / mL saponin solution and 0.1 mL of tyrosinase solution (activity of 200 U / mL) were added, and mixed. The absorbance change within 0-5 min was measured at 1 min intervals at a wavelength of 475 nm using an ELISA reader, and the IC50 value was calculated.

[0120] For detailed results, see Figure 2As can be seen from the figure, with the increase of saponin mass concentration, the inhibitory effect was significantly enhanced, and the IC50 value was 1.508 mg / mL.

[0121] Example 7

[0122] This example provides a test for the application effect of golden camellia saponin extract in repairing photoaging damage

[0123] Hacat in the growth phase was implanted into a 6-well plate and cultured in an incubator for 24 h. The supernatant was discarded and culture medium containing golden camellia saponins and golden camellia water extract was added to the culture medium. After 2 h, UVB irradiation of 300 mJ / cm 2 , lasting 15 minutes, and set up a blank group and a model group, and added equal volumes of culture medium to the two groups. The blank group was not irradiated, and the model group was irradiated. The IL-1, IL-6 and MDA levels were detected by the kit method.

[0124] For detailed results, please see Figure 3 The results showed that golden camellia saponins can significantly reduce the levels of IL-1, IL-6, and MDA, and repair skin inflammation and oxidative damage caused by photoaging. In addition, the results showed that under equal dosages, its effect is better than that of golden camellia tea water extract.

[0125] Example 8

[0126] This embodiment provides a detection method for the inhibition of melanoma cells by extracts of golden camellia saponins

[0127] A375 melanoma cells in the growth phase were implanted into a 96-well plate and cultured in an incubator for 24 hours. The supernatant was discarded and culture medium containing Camellia chrysantha saponins and Camellia chrysantha water extract was added to the culture medium. After 24 hours, the activity of melanoma cells was detected by CCK-8, and a blank group was set up, to which an equal volume of culture medium was added.

[0128] For detailed results, please see Figure 4 The results showed that golden camellia saponins can reduce the proliferation activity of A375 melanoma cells, and at equal doses, their effect is better than golden camellia saponins water extract.

[0129] Example 9

[0130] This example provides a test for the effect of golden camellia saponin extract on promoting the absorption and penetration of polypeptides

[0131] The ex vivo skin was thawed at room temperature, cut into appropriate size (effective skin area of ​​0.636 cm2), fixed between the supply pool and the receiving pool, with the stratum corneum facing the supply pool, placed in the receiving solution [0.9% saline-ethanol (4:1) mixture], 0.5 g of the golden camellia tea pearl cream prepared in Example 6 was applied to the skin surface, the time was recorded as 0h, 200 μl of samples were taken at 0.5, 1, 2, 4, 6, 8, 10, and 12h, and an equal volume of fresh receiving solution was added in time. The polypeptide content was detected by the biuretic method. The cream without golden camellia tea saponin was used as a control.

[0132] For detailed results, please see Figure 5 The results showed that adding golden camellia saponin extract can significantly increase the skin permeability of peptides.

[0133] In summary, the present application develops the application of golden camellia saponins in the field of cosmetics, and further in the field of whitening; among them, it has been tested that the extraction purity of golden camellia saponins extracted by the extraction method of golden camellia saponin extract provided by the present application can be as high as 83%; by further comparing the water extract obtained by water extraction for melanoma cell activity inhibition test, under the condition of equal dose, the inhibition rate of golden camellia saponin extract on melanoma cell activity is significantly higher than that of water extract; in addition, the present application further finds that golden camellia saponins have a more significant effect on promoting the absorption and penetration of effective ingredients, such as polypeptide absorption. The test results show that the addition of golden camellia saponins can significantly increase the skin permeability of polypeptides, so it has good application prospects in the field of cosmetics. The present application fills the gap in golden camellia related extracts in this field, and provides a good research basis and reference for subsequent in-depth research.

[0134] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application may be combined with each other.

[0135] The above description is only a preferred embodiment of the present application and does not constitute any form of limitation to the present application. Any simple modification, equivalent changes and modifications made to the above embodiments based on the technical essence of the present application are within the scope of the technical solution of the present application.

Claims

1. A method for extracting golden camellia saponin extract, characterized in that: The extraction method comprises the following steps: S1. Preheat the ultrasonic cleaning machine to 45-55°C, put the golden camellia tea powder into a conical flask, and add ethanol; S2. Add an appropriate amount of 0.1 mol / L dilute hydrochloric acid to adjust the pH to 4, then add cellulase and shake well; S3, place the conical flask in an ultrasonic cleaning machine, extract under ultrasound for 45-55 minutes, take the macroporous resin wet method column, and load the sample; S4. After adsorption for 24 hours, the mixture was eluted with 75% ethanol, the effluent was collected, and evaporated to dryness in a water bath to obtain the golden camellia saponin extract.

2. The extraction method according to claim 1, characterized in that: In step S2, the volume fraction of ethanol is 55%-58%; 4%-4.5% of cellulase is added; In step S3, the ultrasonic power is 400 W; the macroporous resin is 10 g of D101 macroporous resin, and the flow rate of the extract during loading is 0.5 mL min -1 , volume 1BV; In step S4, 1BV of liquid is discharged.

3. A delivery system comprising a saponin extract of Camellia chrysantha, characterized in that: The golden camellia saponin extract is obtained by the extraction method according to claim 1 or 2; The delivery systems include liposomes, cyclodextrins, Pickering emulsions, and hydrogels.

4. A method for preparing golden camellia saponin liposomes, characterized in that: The preparation method comprises the following steps: Weigh golden camellia tea saponin, add PBS buffer with a pH of 7.5-8.5, and ultrasonically dissolve for 4-6 minutes before use; the golden camellia tea saponin is extracted by the extraction method according to claim 1 or 2; Weigh lecithin and cholesterol, dissolve them in methanol in a round-bottom flask, and evaporate them at 40-50°C until a uniform film forms; The mass ratio of the golden camellia saponin, the lecithin and the cholesterol is 2:4:

1.

5. A method for preparing golden camellia saponin cyclodextrin, characterized in that: The preparation method comprises the following steps: Weighing cyclodextrin and golden camellia saponin, placing the cyclodextrin in a culture dish, and adding water to fully dissolve it; the golden camellia saponin is extracted by the extraction method according to claim 1 or 2; Dissolve golden camellia saponin in water, then slowly drop the golden camellia saponin solution into the cyclodextrin solution, stir, centrifuge the obtained solution at 10°C and 14000r / min for 1h, remove the supernatant, and then freeze in a freeze dryer until completely dry to obtain a white solid powder; The mass ratio of the cyclodextrin to the golden camellia tea is 2:1, the inclusion time is 2.5 hours, and the inclusion temperature is 4°C.

6. A golden camellia saponin pearl cream, characterized in that: Includes the following components: Phase A: Camellia seed oil 3%, isononyl isononanoate 5%, caprylic / capric triglyceride 3%, bisabolol 0.2%, tocopheryl acetate 0.5%, lemon essential oil 0.1%, emulsifier A165 1.89%, emulsifier M82 1.11%; Item B: 3.5% glycerol, 2% propylene glycol, 1.5% trehalose, 2% 1-3 propanediol, 0.35% carbomer U10, 0.1% β-glucan, 0.1% glabridin, 0.02% EDTA-2Na, add water to 100%; Phase C: 0.5% PEG-40 hydrogenated castor oil, 4% pearl hydrolyzate, 0.5% phenoxyethanol, 0.05% golden camellia saponin; the golden camellia saponin is extracted by the extraction method according to claim 1 or 2.

7. The golden camellia tea saponin pearl cream according to claim 6, characterized in that: The golden camellia saponin pearl cream is prepared by the following method: The components of the phase A, the phase B, and the phase C are mixed and placed in different beakers for uniform dispersion and standby use; Take 2g of sodium hydroxide solid in a conical flask, add 18g of ultrapure water to make a 10% sodium hydroxide solution, which is used as the acid-base regulator in this experiment; Place the beaker of phase C in a constant temperature water bath at 55-65°C, stir and dissolve until uniform, and set aside; Add the phase A into the beaker of the phase B, turn on the homogenizer, and homogenize for 5 to 10 minutes at a speed of 3000 r / min; Add acid-base regulator to adjust pH value and continue homogenizing and emulsifying for 2-4 minutes; When the cream is cooled to 40-60℃ and kept for more than 30 minutes, add the evenly mixed phase C to the cream, homogenize and emulsify at 3000r / min for 3-5 minutes, stir evenly and keep warm for 20 minutes to obtain the base cream.

8. Use of golden camellia saponin extracted by the extraction method according to any one of claims 1-2, or golden camellia saponin liposome prepared by the preparation method according to claim 4, or golden camellia saponin cyclodextrin prepared by the preparation method according to claim 5, or golden camellia saponin pearl cream according to any one of claims 6-7 in promoting the absorption and penetration of effective ingredients.

9. Use of golden camellia saponin extracted by the extraction method according to any one of claims 1-2, or golden camellia saponin liposome prepared by the preparation method according to claim 4, or golden camellia saponin cyclodextrin prepared by the preparation method according to claim 5, or golden camellia saponin pearl cream according to any one of claims 6-7 in inhibiting tyrosinase activity.

10. Use of golden camellia saponin extracted by the extraction method according to any one of claims 1-2, or golden camellia saponin liposome prepared by the preparation method according to claim 4, or golden camellia saponin cyclodextrin prepared by the preparation method according to claim 5, or golden camellia saponin pearl cream according to any one of claims 6-7 in repairing photoaging damage caused by ultraviolet radiation.

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