Preparation method of camellia yunnanensis flower extract, product and application thereof

Abstract

This invention relates to a method for preparing an extract of Camellia yunnanensis flower, its product, and its applications. The preparation method includes: mixing Camellia yunnanensis flower raw material with water and an enzyme, performing enzymatic hydrolysis, and then inactivating the enzyme; subjecting the enzymatic hydrolysis solution to ultrasonic treatment, performing solid-liquid separation, concentrating and drying the supernatant to obtain a crude extract; mixing the crude extract with an aqueous two-phase extractant for extraction, concentrating and drying the supernatant to obtain the Camellia yunnanensis flower extract. This extraction process can maximize the yield of extracts with tyrosinase inhibitory activity at lower temperatures; the use of aqueous two-phase extraction enriches multiple target active ingredients, ensuring that the extracted product has superior tyrosinase inhibitory activity. This invention also develops a formulation with excellent stability containing the aforementioned Camellia yunnanensis flower extract, wherein the Camellia yunnanensis flower extract is encapsulated in cyclodextrin, the inclusion complex is loaded in a liposome structure, and the addition of Camellia yunnanensis seed oil and Prickly pear fruit oil can further improve the stability of the obtained formulation.

Description

Technical Field

[0001] This invention belongs to the field of plant extraction technology, and relates to a method for preparing Yunnan camellia flower extract, its products, and applications. Background Technology

[0002] Skin color is primarily determined by the amount and distribution of skin pigment, with melanin being the most significant factor. Melanocytes in the skin produce melanin, and melanin granules are transferred to keratinocytes via the dendritic processes of melanocytes. The melanin granules transferred to keratinocytes then travel upwards with the epidermal cells to the stratum corneum, thus affecting skin color or forming age spots.

[0003] Currently, most skin whitening agents work by inhibiting tyrosinase activity or blocking the synthesis pathway of melanin from tyrosine, thereby reducing melanin production and achieving a whitening effect. These skin whitening agents mainly include arbutin, hydroquinone and its derivatives, L-ascorbic acid and its derivatives, kojic acid and its derivatives, vitamin C ethyl ether and its derivatives, niacinamide, tranexamic acid, hydroquinone and its derivatives, etc. With the increasing demand from consumers for safe and natural ingredients, highly effective and low-irritant natural tyrosinase inhibitors will become an important development direction for the cosmetics industry.

[0004] Camellia yunnanensis flowers are rich in polyphenolic compounds and water-soluble pigments, among other active ingredients. These components possess tyrosinase inhibitory activity, effectively blocking the synthesis of melanin both in vivo and in vitro, making them natural and highly effective whitening active ingredients. However, there are few reports in existing technologies on how to extract Camellia yunnanensis flowers to obtain extracts with higher tyrosinase inhibitory activity. Furthermore, the color stability of Camellia yunnanensis flower extracts is significantly affected by the chemical composition and structure, as well as environmental factors (such as light, heat, pH, and oxygen), which limits its application in cosmetic formulations. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a method for preparing Camellia yunnanensis extract, its products and applications, specifically involving a method for preparing Camellia yunnanensis extract with tyrosinase inhibitory activity and whitening effect, a highly stable preparation containing the aforementioned Camellia yunnanensis extract and its preparation method, and the application of the aforementioned Camellia yunnanensis extract or preparation in the preparation of tyrosinase inhibitors or products with whitening effect.

[0006] To achieve this objective, the present invention adopts the following technical solution:

[0007] In a first aspect, the present invention provides a method for preparing Camellia yunnanensis flower extract, the method comprising the following steps:

[0008] (1) Mix the Yunnan camellia flower raw material with water and enzymes, perform enzymatic hydrolysis, and then inactivate the enzyme to obtain an enzymatic hydrolysis solution; perform ultrasonic treatment on the enzymatic hydrolysis solution, then separate the solid and liquid, concentrate and dry the clear liquid to obtain the crude extract of Yunnan camellia flower;

[0009] (2) The crude extract of Camellia yunnanensis was mixed with a two-phase extractant for extraction. The upper phase after extraction was concentrated and dried to obtain the Camellia yunnanensis extract.

[0010] The upper phase of the aqueous two-phase extractant is an aqueous solution containing ethanol, and the lower phase of the aqueous two-phase extractant is an aqueous solution containing water-soluble salts.

[0011] This invention creatively employs enzymatic hydrolysis combined with ultrasonic treatment to extract Yunnan camellia flower raw materials, enabling the extraction of extracts with tyrosinase inhibitory activity to be maximized at lower temperatures. Compared with single enzymatic hydrolysis, single ultrasonic extraction, or alcohol extraction, the resulting product exhibits superior tyrosinase inhibitory activity. Furthermore, an aqueous two-phase extraction method is used to enrich multiple target active ingredients instead of traditional column chromatography purification, ensuring that the extracted product has even better tyrosinase inhibitory activity.

[0012] Preferably, the enzyme includes pectinase and / or cellulase.

[0013] A more preferred combination is pectinase and cellulase.

[0014] This invention discovers that using a combination of pectinase and cellulase to enzymatically hydrolyze Camellia yunnanensis raw materials results in an extract with superior tyrosinase inhibitory activity compared to other enzymatic hydrolysis methods.

[0015] Preferably, the mass ratio of pectinase to cellulase is (1-3):1, for example, 1:1, 1.2:1, 1.5:1, 1.8:1, 2:1, 2.2:1, 2.5:1, 2.8:1, 3:1, etc.

[0016] Preferably, the amount of enzyme added is 0.1-1% of the enzymatic hydrolysis system, such as 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, etc.

[0017] Preferably, the mass ratio of the Yunnan camellia flower raw material to water is 1:(5-20), such as 1:5, 1:7, 1:8, 1:9, 1:10, 1:12, 1:13, 1:14, 1:16, 1:18, 1:20, etc.

[0018] Preferably, the pH value of the enzymatic hydrolysis system is 4-7, such as pH=4, pH=4.5, pH=5, pH=5.5, pH=6, pH=6.5, pH=7, etc.; the temperature of the enzymatic hydrolysis treatment is 30-50℃, such as 30℃, 35℃, 40℃, 45℃, 50℃, etc.; and the time of the enzymatic hydrolysis treatment is 30-60 min, such as 30 min, 35 min, 40 min, 45 min, 50 min, 55 min, 60 min, etc.

[0019] Preferably, the power of the ultrasonic treatment is 100-400 W, such as 100 W, 150 W, 200 W, 250 W, 300 W, 350 W, 400 W, etc.; the ultrasonic treatment time is 30-60 min, such as 30 min, 40 min, 50 min, 60 min, etc.; and the ultrasonic treatment temperature is 30-50℃, such as 30℃, 35℃, 40℃, 45℃, 50℃, etc.

[0020] Preferably, the concentration in step (1) is carried out by vacuum concentration.

[0021] Preferably, the drying in step (1) is performed by freeze drying.

[0022] Preferably, the upper phase of the aqueous two-phase extractant is an aqueous solution containing 20-30 wt% (e.g., 20 wt%, 22 wt%, 23 wt%, 24 wt%, 26 wt%, 28 wt%, 30 wt%, etc.) of ethanol.

[0023] Preferably, the lower phase of the aqueous two-phase extractant is an aqueous solution containing 10-20% (e.g., 10%, 12%, 14%, 15%, 16%, 18%, 20%, etc.) of water-soluble salt.

[0024] This invention employs an upper aqueous phase with a specific alcohol concentration and a lower aqueous phase with a specific salt concentration for crude extraction, which can obtain the active ingredient with the optimal tyrosinase inhibitory effect.

[0025] Preferably, the water-soluble salt includes any one or a combination of at least two of sodium citrate, potassium citrate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, or ammonium sulfate; more preferably, a combination of sodium citrate and ammonium sulfate.

[0026] This invention also creatively discovers that selecting the combination of sodium citrate and ammonium sulfate as the salt in the lower aqueous phase results in better extraction and superior product quality compared to other water-soluble salt components.

[0027] Preferably, the mass ratio of sodium citrate to ammonium sulfate is (1-3):1, such as 1:1, 1.2:1, 1.5:1, 1.8:1, 2:1, 2.2:1, 2.5:1, 2.8:1, 3:1, etc.

[0028] Preferably, the aqueous two-phase extractant system further uses citric acid to adjust the pH value to 5.0-7.0, such as pH=5.0, pH=5.5, pH=6.0, pH=6.5, pH=7.0, etc.

[0029] Preferably, the ratio of the crude extract of Camellia yunnanensis to the aqueous two-phase extractant is 1:(5-20) g / mL, for example, 1:5 g / mL, 1:7 g / mL, 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, 1:11 g / mL, 1:12 g / mL, 1:13 g / mL, 1:14 g / mL, 1:15 g / mL, etc.

[0030] Preferably, the extraction process is carried out at 30-50℃ (e.g., 30℃, 35℃, 40℃, 45℃, 50℃, etc.) and stirred for 30-60 min (e.g., 30 min, 40 min, 50 min, 60 min, etc.) before being allowed to stand and separate into layers.

[0031] Preferably, the concentration in step (2) is carried out by vacuum concentration.

[0032] Preferably, the drying in step (2) is performed by freeze drying.

[0033] In a second aspect, the present invention provides a Camellia yunnanensis flower extract prepared according to the preparation method described in the first aspect.

[0034] Thirdly, the present invention provides a preparation containing Camellia yunnanensis flower extract, wherein the raw materials for preparing the preparation include the following components: lecithin, phytosterols, Camellia yunnanensis seed oil, Prickly pear fruit oil, the Camellia yunnanensis flower extract described in the second aspect, and cyclodextrin.

[0035] To overcome the drawback of the color instability of Camellia yunnanensis extract, which limits its application in cosmetic formulations, this invention also develops a formulation with excellent stability containing the aforementioned Camellia yunnanensis flower extract. In this formulation, the Camellia yunnanensis flower extract is encapsulated in cyclodextrin, and the inclusion complex is then loaded into a liposome structure. The addition of Camellia yunnanensis seed oil and Prickly pear fruit oil further enhances the stability of the formulation, and both have a synergistic effect in improving stability.

[0036] Preferably, the lecithin includes any one or a combination of at least two of soybean lecithin, egg yolk lecithin, hydrogenated lecithin, dipalmitoylphosphatidylcholine, or dioleoylphosphatidylcholine.

[0037] Preferably, the phytosterols include any one or a combination of at least two of β-sitosterol, stigmasterol, campesterol, or sitosterol.

[0038] Preferably, the cyclodextrin comprises hydroxypropyl-β-cyclodextrin.

[0039] Preferably, the raw materials for preparing the preparation include the following components by mass fraction: 35-45% lecithin, 2-4% phytosterols, 2-4% Camellia yunnanensis seed oil, 1-4% Citrus aurantium fruit oil, 6-8% Camellia yunnanensis flower extract as described in the second aspect, and 12-16% cyclodextrin.

[0040] The mass fraction of the lecithin can be selected from 35%, 38%, 40%, 42%, 43%, 44%, 45%, etc.; the mass fraction of the phytosterols can be selected from 2%, 2.5%, 3%, 3.5%, 4%, etc.; the mass fraction of the Camellia yunnanensis seed oil can be selected from 2%, 2.5%, 3%, 3.5%, 4%, etc.; the mass fraction of the Prickly pear fruit oil can be selected from 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, etc.; the mass fraction of the Camellia yunnanensis flower extract can be selected from 6%, 6.5%, 7%, 7.5%, 8%, etc.; the mass fraction of the cyclodextrin can be selected from 12%, 12.5%, 13%, 14%, 14.5%, 15%, 16%, etc.

[0041] Preferably, the raw materials for preparing the formulation further include a freeze-drying protectant, including any one or a combination of at least two of trehalose, sucrose, mannose, mannitol, sorbitol, glycerol, polyvinylpyrrolidone, or polyethylene glycol.

[0042] Fourthly, the present invention provides a method for preparing a formulation containing Camellia yunnanensis extract according to the third aspect, the preparation method comprising the following steps:

[0043] (1) Dissolve the Yunnan camellia flower extract in a buffer solution and then mix it with cyclodextrin to obtain solution A;

[0044] Lecithin, phytosterols, Camellia yunnanensis seed oil, and Prickly pear fruit oil were dissolved in an ethanol solution and then evaporated to obtain a lipid film.

[0045] (2) Solution A is added to the lipid film for hydration treatment, and then homogenized to obtain the preparation containing the Yunnan camellia extract.

[0046] Preferably, the pH of the buffer solution is 4-7, such as pH=4, pH=4.5, pH=5, pH=5.5, pH=6, pH=6.5, pH=7, etc.

[0047] When the pH of the buffer solution used is in the range of 4-7, the resulting formulation has better stability.

[0048] Preferably, the mixing and stirring is carried out at 20-40℃ (e.g., 20℃, 25℃, 30℃, 35℃, 40℃, etc.) for 2-6 hours (e.g., 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, etc.).

[0049] Preferably, the hydration treatment is carried out at 40-60℃ (e.g., 40℃, 45℃, 50℃, 55℃, 60℃, etc.) for 1-3 hours (e.g., 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, etc.).

[0050] Preferably, the homogenization process is performed 1 to 5 cycles (e.g., 1, 2, 3, 4 or 5) at 8000-10000 psi (e.g. 8000 psi, 8500 psi, 9000 psi, 9500 psi, 10000 psi, etc.), each cycle lasting 2 to 5 minutes (e.g., 2 minutes, 3 minutes, 4 minutes, 5 minutes, etc.).

[0051] Preferably, the homogenized product is further subjected to freeze-drying after homogenization.

[0052] Fifthly, the present invention provides the use of the Camellia yunnanensis extract described in the second aspect or the preparation containing the Camellia yunnanensis extract described in the third aspect in the preparation of products with whitening effects.

[0053] In a sixth aspect, the present invention provides the use of the Camellia yunnanensis extract described in the second aspect or the formulation containing the Camellia yunnanensis extract described in the third aspect in the preparation of tyrosinase inhibitors.

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

[0055] This invention creatively employs enzymatic hydrolysis combined with ultrasonic treatment to extract Yunnan camellia flower raw materials, enabling the extraction of extracts with tyrosinase inhibitory activity to be maximized at lower temperatures. Compared with single enzymatic hydrolysis, single ultrasonic extraction, or alcohol extraction, the resulting product exhibits superior tyrosinase inhibitory activity. Furthermore, an aqueous two-phase extraction method is used to enrich multiple target active ingredients instead of traditional column chromatography purification, ensuring that the extracted product has even better tyrosinase inhibitory activity.

[0056] To overcome the drawback of the color instability of Camellia yunnanensis extract, which limits its application in cosmetic formulations, this invention also develops a formulation with excellent stability containing the aforementioned Camellia yunnanensis flower extract. In this formulation, the Camellia yunnanensis flower extract is encapsulated in cyclodextrin, and the inclusion complex is then loaded into a liposome structure. The addition of Camellia yunnanensis seed oil and Prickly pear fruit oil further enhances the stability of the resulting formulation, and both have a synergistic effect in improving stability. Attached Figure Description

[0057] Figure 1 This is a liquid chromatogram of the Camellia yunnanensis extract prepared in Example 1. Detailed Implementation

[0058] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0059] The pectinase, cellulase, phytosterols (a mixture of campesterol, stigmasterol, and β-sitosterol) mentioned below were purchased from Nanning Pangbo Biotechnology Co., Ltd.; the cellulase was purchased from Nanning Pangbo Biotechnology Co., Ltd.; the phytosterols were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; the camellia seed oil was from Yunnan Specialty Plant Extraction Laboratory Co., Ltd.; the prickly ash fruit oil was from Yunnan Specialty Plant Extraction Laboratory Co., Ltd.; and the hydroxypropyl-β-cyclodextrin was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0060] Preparation Example 1

[0061] This preparation example provides a Camellia yunnanensis flower extract, which is prepared by the following method:

[0062] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 10 times the mass of water and enzyme (pectinase and cellulase in a mass ratio of 2:1, which is 0.5% of the enzymatic hydrolysis system). The pH of the solvent was adjusted to 5 with citric acid and enzymatic hydrolysis was carried out at 40℃ for 45 min. After the enzymatic hydrolysis was completed, the enzyme was inactivated at 75℃ for 10 min to obtain the enzymatic hydrolysis solution.

[0063] (2) The enzymatic hydrolysate was subjected to ultrasonic treatment at a power of 200 W for 45 min and a temperature of 40℃; then the solution was filtered under reduced pressure, the filtrate was concentrated under reduced pressure, and freeze-dried to obtain crude extract of Camellia yunnanensis.

[0064] (3) The crude extract of Camellia yunnanensis was mixed with the aqueous two-phase extractant at a ratio of 1:10 g / mL, stirred at 40℃ for 40 min, allowed to stand and separated, and the upper phase after extraction was concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract. The upper phase of the aqueous two-phase extractant was an aqueous solution containing 20wt% ethanol, and the lower phase of the aqueous two-phase extractant was an aqueous solution containing 10% water-soluble salt (sodium citrate and ammonium sulfate in a mass ratio of 3:2), and the pH was adjusted to 6.0 using citric acid.

[0065] Preparation Example 2

[0066] This preparation example provides a Camellia yunnanensis flower extract, which is prepared by the following method:

[0067] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 15 times the mass of water and enzyme (pectinase and cellulase in a mass ratio of 3:1, which is 0.8% of the enzymatic hydrolysis system). The pH of the solvent was adjusted to 6 with citric acid and enzymatic hydrolysis was carried out at 45℃ for 40 min. After the enzymatic hydrolysis was completed, the enzyme was inactivated at 75℃ for 10 min to obtain the enzymatic hydrolysis solution.

[0068] (2) The enzymatic hydrolysate was subjected to ultrasonic treatment at a power of 300 W for 40 min and a temperature of 50℃; then the solution was filtered under reduced pressure, the filtrate was concentrated under reduced pressure, and freeze-dried to obtain crude extract of Camellia yunnanensis.

[0069] (3) The crude extract of Camellia yunnanensis was mixed with the aqueous two-phase extractant at a ratio of 1:15 g / mL, stirred at 45℃ for 50 min, allowed to stand and separated, and the upper phase after extraction was concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract. The upper phase of the aqueous two-phase extractant was an aqueous solution containing 25wt% ethanol, and the lower phase of the aqueous two-phase extractant was an aqueous solution containing 15% water-soluble salt (potassium citrate and disodium hydrogen phosphate in a mass ratio of 2:1), and the pH was adjusted to 7.0 with citric acid.

[0070] Preparation Example 3

[0071] This preparation example provides a Camellia yunnanensis flower extract, which is prepared by the following method:

[0072] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 8 times the mass of water and enzyme (pectinase and cellulase in a mass ratio of 1:1, which is 0.3% of the enzymatic hydrolysis system). The pH of the solvent was adjusted to 5.5 with citric acid and enzymatic hydrolysis was carried out at 35℃ for 50 min. After the enzymatic hydrolysis was completed, the enzyme was inactivated at 75℃ for 10 min to obtain the enzymatic hydrolysis solution.

[0073] (2) The enzymatic hydrolysate was subjected to ultrasonic treatment at a power of 250 W for 50 min and a temperature of 45℃; then the solution was filtered under reduced pressure, the filtrate was concentrated under reduced pressure, and freeze-dried to obtain crude extract of Camellia yunnanensis.

[0074] (3) The crude extract of Camellia yunnanensis was mixed with the aqueous two-phase extractant at a ratio of 1:8 g / mL, stirred at 35°C for 60 min, allowed to stand and separated, and the upper phase after extraction was concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract. The upper phase of the aqueous two-phase extractant was an aqueous solution containing 30 wt% ethanol, and the lower phase of the aqueous two-phase extractant was an aqueous solution containing 10% water-soluble salt (dipotassium hydrogen phosphate and ammonium sulfate in a mass ratio of 3:2), and the pH was adjusted to 5.5 with citric acid.

[0075] Preparation Example 4

[0076] This preparation example provides a Yunnan camellia flower extract, the preparation method of which differs from that of preparation example 1 only in that the enzyme used in step (1) is a single pectinase, and the total amount of enzyme added remains unchanged, while other operations remain unchanged.

[0077] Preparation Example 5

[0078] This preparation example provides a Yunnan camellia flower extract, the only difference between its preparation method and that of Preparation Example 1 is that the enzyme used in step (1) is a single cellulase, and the total amount of enzyme added remains unchanged, while other operations remain unchanged.

[0079] Preparation Example 6

[0080] This preparation example provides a Yunnan camellia flower extract, the preparation method of which differs from that of preparation example 1 only in that: in step (3), the upper phase of the aqueous two-phase extractant is an aqueous solution containing 10wt% ethanol, and all other operations remain unchanged.

[0081] Preparation Example 7

[0082] This preparation example provides a Yunnan camellia flower extract, the preparation method of which differs from that of Preparation Example 1 only in that: in step (3), the upper phase of the aqueous two-phase extractant is an aqueous solution containing 40wt% ethanol, and all other operations remain unchanged.

[0083] Preparation Example 8

[0084] This preparation example provides a Yunnan camellia flower extract, the preparation method of which differs from that of Preparation Example 1 only in that: in step (3), the lower phase of the aqueous two-phase extractant is an aqueous solution containing 30% water-soluble salt (sodium citrate and ammonium sulfate in a mass ratio of 3:2), and all other operations remain unchanged.

[0085] Preparation Example 9

[0086] This preparation example provides a Yunnan camellia flower extract, the preparation method of which differs from that of Preparation Example 1 only in that: in step (3), the lower phase of the aqueous two-phase extractant is an aqueous solution containing 10% water-soluble salt (sodium citrate), and all other operations remain unchanged.

[0087] Preparation Example 10

[0088] This preparation example provides a Yunnan camellia flower extract, the preparation method of which differs from that of Preparation Example 1 only in that: in step (3), the lower phase of the aqueous two-phase extractant is an aqueous solution containing 10% water-soluble salt (ammonium sulfate), and all other operations remain unchanged.

[0089] Comparative Preparation Example 1

[0090] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1 is that the enzymatic hydrolysis solution is not subjected to ultrasonic treatment, and the enzymatic hydrolysis time is extended, as detailed below:

[0091] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 10 times the mass of water and enzymes (pectinase and cellulase in a mass ratio of 2:1, which accounted for 0.5% of the enzymatic hydrolysis system). The pH of the solvent was adjusted to 5 with citric acid and enzymatic hydrolysis was carried out at 40℃ for 90 min. After the enzymatic hydrolysis was completed, the enzymes were inactivated at 75℃ for 10 min to obtain the enzymatic hydrolysis solution. Then, the solution was filtered under reduced pressure, the filtrate was concentrated under reduced pressure, and freeze-dried to obtain the crude extract of Camellia yunnanensis flowers.

[0092] (2) Same as step (3) of preparation example 1.

[0093] Comparative Preparation Example 2

[0094] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1. Instead of enzymatic hydrolysis of the raw materials, only ultrasonic treatment is performed, and the ultrasonic treatment time is extended, as detailed below:

[0095] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 10 times the mass of water. The mixture was subjected to ultrasonic treatment at a power of 200 W for 90 min and a temperature of 40℃. The mixture was then filtered under reduced pressure, the filtrate was concentrated under reduced pressure, and freeze-dried to obtain crude extract of Camellia yunnanensis flowers.

[0096] (2) Same as step (3) of preparation example 1.

[0097] Comparative preparation example 3

[0098] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1, except that the raw materials are subjected to enzymatic hydrolysis and ultrasonic treatment simultaneously, as detailed below:

[0099] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 10 times the mass of water and enzymes (pectinase and cellulase in a mass ratio of 2:1, which accounted for 0.5% of the enzymatic hydrolysis system). The pH of the solvent was adjusted to 5 with citric acid and enzymatic hydrolysis was carried out at 40°C for 45 min. At the same time, ultrasonic treatment was carried out at a power of 200 W for 45 min. After the enzymatic hydrolysis was completed, the enzyme was inactivated at 75°C for 10 min to obtain the enzymatic hydrolysis solution. Then, the solution was filtered under reduced pressure, the filtrate was concentrated under reduced pressure, and freeze-dried to obtain the crude extract of Camellia yunnanensis flowers.

[0100] (2) Same as step (3) of preparation example 1.

[0101] Comparative preparation example 4

[0102] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1, except that the raw materials are first subjected to ultrasonic treatment followed by enzymatic hydrolysis, as detailed below:

[0103] (1) Powder 100 g of Yunnan camellia flowers and mix them with 10 times the weight of water. Then, perform ultrasonic treatment. The ultrasonic treatment power is 200 W, the time is 45 min, and the ultrasonic treatment temperature is 40℃.

[0104] (2) Add enzyme (pectinase and cellulase in a mass ratio of 2:1, which is 0.5% of the enzymatic hydrolysis system) to the treated mixture, adjust the pH of the solvent to 5 with citric acid, and carry out enzymatic hydrolysis at 40℃ for 45 min. After the enzymatic hydrolysis is completed, inactivate the enzyme at 75℃ for 10 min to obtain the enzymatic hydrolysis solution; then filter under reduced pressure, concentrate the filtrate under reduced pressure, freeze dry, and obtain crude extract of Camellia yunnanensis.

[0105] (3) Same as step (3) in preparation example 1.

[0106] Comparative preparation example 5

[0107] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1, except that the raw materials are not subjected to enzymatic hydrolysis and ultrasonic treatment, but instead undergo water extraction and reflux treatment, as detailed below:

[0108] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 10 times the weight of water, and heated under reflux for 90 min; then filtered under reduced pressure, the filtrate was concentrated under reduced pressure and freeze-dried to obtain crude extract of Camellia yunnanensis flowers.

[0109] (2) The crude extract of Camellia yunnanensis was mixed with the aqueous two-phase extractant at a ratio of 1:10 g / mL, stirred at 40℃ for 40 min, allowed to stand and separated, and the upper phase after extraction was concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract. The upper phase of the aqueous two-phase extractant was an aqueous solution containing 20wt% ethanol, and the lower phase of the aqueous two-phase extractant was an aqueous solution containing 10% water-soluble salt (sodium citrate and ammonium sulfate in a mass ratio of 3:2), and the pH was adjusted to 6.0 with citric acid.

[0110] Comparative preparation example 6

[0111] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1, except that the raw materials are not subjected to enzymatic hydrolysis and ultrasonic treatment, but instead undergo alcohol extraction and reflux treatment, as detailed below:

[0112] (1) 100 g of Camellia yunnanensis flowers were crushed and mixed with 10 times the mass of 70% ethanol aqueous solution, and heated under reflux for 90 min; then filtered under reduced pressure, the filtrate was concentrated under reduced pressure and freeze-dried to obtain crude extract of Camellia yunnanensis flowers;

[0113] (2) The crude extract of Camellia yunnanensis was mixed with the aqueous two-phase extractant at a ratio of 1:10 g / mL, stirred at 40℃ for 40 min, allowed to stand and separated, and the upper phase after extraction was concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract. The upper phase of the aqueous two-phase extractant was an aqueous solution containing 20wt% ethanol, and the lower phase of the aqueous two-phase extractant was an aqueous solution containing 10% water-soluble salt (sodium citrate and ammonium sulfate in a mass ratio of 3:2), and the pH was adjusted to 6.0 with citric acid.

[0114] Comparative preparation example 7

[0115] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1, except that resin elution is used instead of aqueous two-phase extraction in step (3), as follows:

[0116] (3) The crude extract of Camellia yunnanensis was eluted and purified using an AB-8 resin column. First, it was eluted with water for 5 BV, and then eluted with 60% ethanol aqueous solution. The eluent of 60% ethanol aqueous solution was collected, concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract.

[0117] Comparative Preparation Example 8

[0118] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1, except that resin elution is used instead of aqueous two-phase extraction in step (3), as follows:

[0119] (3) The crude extract of Camellia yunnanensis was eluted and purified using an AB-8 resin column. First, 5 BV of water was used to elute, then 4 BV of 30% ethanol aqueous solution was used to elute, and then 60% ethanol aqueous solution was used to elute. The eluent of 60% ethanol aqueous solution was collected, concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract.

[0120] Comparative preparation example 9

[0121] This preparation example provides a Camellia yunnanensis flower extract, the only difference between which is the preparation method and that of Preparation Example 1, except that resin elution is used instead of aqueous two-phase extraction in step (3), as follows:

[0122] (3) The crude extract of Camellia yunnanensis was eluted and purified using an AB-8 resin column. First, it was eluted with water for 5 BV, and then eluted with 30% ethanol aqueous solution. The eluent of 30% ethanol aqueous solution was collected, concentrated under reduced pressure and freeze-dried to obtain the Camellia yunnanensis extract.

[0123] Test Example 1

[0124] The contents of proanthocyanidins B2, epicatechin, proanthocyanidins C1, 1,2,3,6-tetra-O-galloyl-β-D-glucose (TeGG), and tyrosinase inhibitory activity of the Camellia yunnanensis flower extracts prepared in Preparation Examples 1-10 and Comparative Preparation Examples 1-9 were evaluated.

[0125] (1) Determination of the contents of proanthocyanidins B2, epicatechin, proanthocyanidins C1, and TeGG: HPLC-DAD (Agilent, 1260 Infinity II) was used for determination. The chromatographic column was an InfinityLab Poroshell 120 EC-C18 (4.6 x 100 mm, 2.7 μM), the injection volume was 5 μL, the column temperature was 25℃, and the mobile phases were A (0.1% formic acid water) and B (acetonitrile). The elution program was: 0 min, 5% B phase; 5 min, 15% B phase; 15 min, 20% B phase; 20 min, 25% B phase; 21 min, 100% B phase; 26 min, 100% B phase. The detection wavelength was 280 nm. The HPLC chromatogram of the Camellia yunnanensis extract prepared in Preparation Example 1 is shown below. Figure 1 As shown.

[0126] (2) Tyrosinase activity test:

[0127] (2.1) Preparation of tyrosinase solution (≥500 U / mg): Accurately weigh 0.01 g, dissolve in 50 mL buffer solution to obtain 100 U / mL tyrosinase solution, and store at 4℃ for later use.

[0128] (2.2) Preparation of L-tyrosine solution: Accurately weigh L-tyrosine, dissolve it in pH 6.8 phosphate buffer solution to prepare a 0.5 mg / mL solution, shake well and store at 4℃ for later use.

[0129] (2.3) Preparation of sample solution: Accurately weigh the Yunnan camellia flower extract and add deionized water to prepare a 1 mg / mL sample to be tested.

[0130] (2.4) Enzyme inhibition activity test: The sample addition reaction was carried out in a 96-well plate, and the sample addition order was as follows:

[0131] Control group B: 100 μL tyrosine solution + 50 μL tyrosinase + 50 μL buffer solution;

[0132] Blank group B0: 100 μL of tyrosine solution + 100 μL of buffer solution;

[0133] Experimental group A: 100 μL tyrosine solution + 50 μL tyrosinase solution + 50 μL samples of different concentrations;

[0134] Color matching group A0: 100 μL tyrosine solution + 50 μL buffer + 50 μL samples of different concentrations;

[0135] After sample addition, the mixture was allowed to react at room temperature for 30 min, and the absorbance (OD) value was measured at 475 nm. The formula for calculating the tyrosinase inhibition rate (IR) is as follows: IR = (1 - (A - A0) / (B - B0)) × 100%. The test results are shown in Table 1.

[0136] Table 1

[0137]

[0138] As shown in Table 1, the Camellia yunnanensis extract prepared by the method of the present invention has excellent tyrosinase inhibitory activity, that is, excellent whitening potential.

[0139] Comparing the data results of Preparation Example 1 with those of Comparative Preparation Examples 1-6, it can be seen that the method of combining enzymatic hydrolysis with subsequent ultrasonic treatment in this invention has superior tyrosinase inhibitory activity compared with single enzymatic hydrolysis, single ultrasonic extraction, ultrasonic followed by enzymatic hydrolysis, simultaneous ultrasonic and enzymatic hydrolysis, alcohol extraction, and water extraction.

[0140] Comparing the data from Preparation Example 1 with those from Comparative Preparation Examples 7-9, it can be seen that the present invention uses a two-phase extraction method for purification instead of the traditional column chromatography purification method, which can ensure that the extracted product has superior tyrosinase inhibitory activity.

[0141] Example 1

[0142] This embodiment provides a formulation whose raw materials are: 40% soybean lecithin, 2% phytosterols, 2% Camellia yunnanensis seed oil, 1% Cirsium japonicum fruit oil, 8% Camellia yunnanensis flower extract of Preparation Example 1, 16% hydroxypropyl-β-cyclodextrin, 25% trehalose, and 6% mannitol.

[0143] The preparation method is as follows:

[0144] (1) Prepare a citrate-disodium hydrogen phosphate buffer solution with pH=5.0. Dissolve the Yunnan camellia flower extract in the buffer solution to prepare a Yunnan camellia flower extract of 10 mg / mL. Add hydroxypropyl-β-cyclodextrin and stir at 25℃ for 4 h to obtain solution A. Let stand for later use.

[0145] (2) Soybean lecithin, phytosterols, Camellia yunnanensis seed oil and Prickly pear fruit oil were dissolved in anhydrous ethanol, evaporated under reduced pressure, and purged with nitrogen until the ethanol was completely removed to obtain a lipid film.

[0146] (3) Dissolve trehalose and mannitol in a citrate-disodium hydrogen phosphate buffer solution at pH 5.0, mix with solution A, and heat and stir at 50°C to obtain solution B;

[0147] (4) Add solution B to the lipid film obtained in step (2) and stir at 50°C for 2 h to obtain solution C;

[0148] (5) The solution C was homogenized under high pressure at 8000 psi for 3 min, and homogenized 3 times in total. The solution was then freeze-dried to obtain the final product.

[0149] Example 2

[0150] This embodiment provides a formulation whose raw materials are: 30% soybean lecithin, 5% hydrogenated lecithin, 4% phytosterols, 3% Camellia yunnanensis seed oil, 2% Citrus aurantium fruit oil, 7% Camellia yunnanensis flower extract of Preparation Example 2, 14% hydroxypropyl-β-cyclodextrin, 25% mannose, and 10% sorbitol.

[0151] The preparation method is as follows:

[0152] (1) Prepare a citrate-disodium hydrogen phosphate buffer solution with pH=6.0. Dissolve the Yunnan camellia flower extract in the buffer solution to prepare a Yunnan camellia flower extract of 10 mg / mL. Add hydroxypropyl-β-cyclodextrin and stir at 30℃ for 3 h to obtain solution A. Let stand for later use.

[0153] (2) Soybean lecithin, hydrogenated lecithin, phytosterols, Camellia yunnanensis seed oil and Prickly pear fruit oil were dissolved in anhydrous ethanol, evaporated under reduced pressure, and purged with nitrogen until the ethanol was completely removed to obtain a lipid film.

[0154] (3) Dissolve mannose and sorbitol in citrate-disodium hydrogen phosphate buffer solution at pH 6.0, mix with solution A, heat and stir at 50°C to obtain solution B;

[0155] (4) Add solution B to the lipid film obtained in step (2) and stir at 55°C for 1 h to obtain solution C;

[0156] (5) The solution C was homogenized under high pressure at 9000 psi for 4 min, and homogenized 3 times in total. The solution was then freeze-dried to obtain the final product.

[0157] Example 3

[0158] This embodiment provides a formulation whose raw materials are: 35% soybean lecithin, 10% hydrogenated lecithin, 2% phytosterols, 4% Camellia yunnanensis seed oil, 4% Citrus aurantium fruit oil, 6% Camellia yunnanensis flower extract of Preparation Example 2, 12% hydroxypropyl-β-cyclodextrin, 20% sucrose, and 7% glycerol.

[0159] The preparation method is as follows:

[0160] (1) Prepare a citrate-disodium hydrogen phosphate buffer solution with pH=6.5. Dissolve the Yunnan camellia flower extract in the buffer solution to prepare a Yunnan camellia flower extract of 10 mg / mL. Add hydroxypropyl-β-cyclodextrin and stir at 35℃ for 2 h to obtain solution A. Let stand for later use.

[0161] (2) Soybean lecithin, hydrogenated lecithin, phytosterols, Camellia yunnanensis seed oil and Prickly pear fruit oil were dissolved in anhydrous ethanol, evaporated under reduced pressure, and purged with nitrogen until the ethanol was completely removed to obtain a lipid film.

[0162] (3) Dissolve sucrose and glycerol in citrate-disodium hydrogen phosphate buffer solution at pH 6.5, mix with solution A, heat and stir at 50°C to obtain solution B;

[0163] (4) Add solution B to the lipid film obtained in step (2) and stir at 45°C for 3 h to obtain solution C;

[0164] (5) The solution C was homogenized under high pressure at 10000 psi for 4 min, and homogenized 3 times in total. The solution was then freeze-dried to obtain the final product.

[0165] Example 4

[0166] This embodiment provides a formulation whose raw materials are: 30% soybean lecithin, 1% phytosterols, 8% Camellia yunnanensis seed oil, 6% Citrus aurantium fruit oil, 8% Camellia yunnanensis flower extract of Preparation Example 1, 16% hydroxypropyl-β-cyclodextrin, 25% trehalose, and 6% mannitol. The preparation method is the same as in Example 1.

[0167] Example 5

[0168] This embodiment provides a formulation whose raw materials are the same as those in Example 1. The only difference in its preparation method is that the pH value of the citrate-disodium hydrogen phosphate buffer solution is 8 in steps (1) and (3), while other conditions remain unchanged.

[0169] Comparative Example 1

[0170] This comparative example provides a formulation whose raw materials differ from those in Example 1 only in that "2% Yunnan camellia seed oil and 1% prickly pear fruit oil" are replaced with "3% Yunnan camellia seed oil," while other components and their contents remain unchanged. The preparation method is the same as in Example 1.

[0171] Comparative Example 2

[0172] This comparative example provides a formulation whose raw materials differ from those in Example 1 only in that "2% Camellia yunnanensis seed oil and 1% Cirsium japonicum fruit oil" are replaced with "3% Cirsium japonicum fruit oil," while other components and their contents remain unchanged. The preparation method is the same as in Example 1.

[0173] Comparative Example 3

[0174] This comparative example provides a formulation whose raw materials differ from those in Example 1 only in that it lacks "2% Camellia yunnanensis seed oil and 1% Cirsium japonicum fruit oil," the content of which is made up by soybean lecithin, while the other components and their contents remain unchanged. The preparation method is the same as in Example 1.

[0175] Comparative Example 4

[0176] This comparative example provides a formulation whose raw materials differ from those in Example 1 only in the absence of "hydroxypropyl-β-cyclodextrin 16%". The reduced amount is proportionally allocated to the other raw materials. The preparation method is as follows:

[0177] (1) Prepare a citrate-disodium hydrogen phosphate buffer solution with pH=5.0. Dissolve the Yunnan camellia flower extract in the buffer solution to prepare a 10 mg / mL Yunnan camellia flower extract solution, and let it stand for later use.

[0178] (2) Soybean lecithin, phytosterols, Camellia yunnanensis seed oil and Prickly pear fruit oil were dissolved in anhydrous ethanol, evaporated under reduced pressure, and purged with nitrogen until the ethanol was completely removed to obtain a lipid film.

[0179] (3) Dissolve trehalose and mannitol in a citrate-disodium hydrogen phosphate buffer solution at pH 5.0, mix with solution A, and heat and stir at 50°C to obtain solution B;

[0180] (4) Add solution B to the lipid film obtained in step (2) and stir at 50°C for 2 h to obtain solution C;

[0181] (5) The solution C was homogenized under high pressure at 8000 psi for 3 min, and homogenized 3 times in total. The solution was then freeze-dried to obtain the final product.

[0182] Application Example 1

[0183] This application example provides a moisturizing essence with whitening effects, the formula of which is shown in Table 2:

[0184] Table 2

[0185]

[0186] The preparation process is as follows: raw materials A, B, and C are premixed evenly, then raw materials D, E, and J are added, and stirred at 85°C for 10 minutes for homogenization at a speed of 6000 rpm for 5 minutes. Stirring is then started, and after cooling to 75°C, raw material F is added and stirred. After cooling to 45°C, raw materials G and H are added and stirred. Then, phase I raw materials are added and stirred until evenly mixed to obtain the final product.

[0187] Application Example 2-5

[0188] Four moisturizing essences are provided. The only difference between the formula and application example 1 is that the preparation obtained in example 1 is replaced with the preparation obtained in examples 2-5 in turn. Other components and contents remain unchanged, and the preparation process is the same as that in application example 1.

[0189] Compare and contrast examples 1-4

[0190] Four moisturizing essences are provided. The only difference between the formula and application example 1 is that the preparation obtained in example 1 is replaced with the preparation obtained in comparative examples 1-4 in turn. Other components and contents remain unchanged, and the preparation process is the same as that in application example 1.

[0191] Comparative Application Example 5

[0192] A moisturizing essence is provided, the only difference between its formulation and application example 1 is that the preparation obtained in example 1 is replaced with the camellia yunnanensis extract obtained in preparation example 1 (the actual amount added is consistent with the camellia yunnanensis extract in example 1), the reduced amount is made up by water, and the other components and contents remain unchanged. Its preparation process is the same as that of application example 1.

[0193] Test Example 2

[0194] Appearance stability test:

[0195] The products obtained from Application Examples 1-5 and Comparative Application Examples 1-5 were stored in the dark at 25°C and 40°C, respectively, and their appearance and color were observed after 30 days. The results are shown in Table 3.

[0196] Table 3

[0197]

[0198] As shown in Table 3, compared with the products of Comparative Application Examples 4-5, the formulation containing Camellia yunnanensis extract developed in this invention exhibits excellent color stability at both room temperature and high temperature by encapsulating Camellia yunnanensis extract with cyclodextrin and then loading the inclusion complex into a liposome structure.

[0199] By comparing the results of Application Example 1 with those of Comparative Application Examples 1-3, it can be seen that the addition of Camellia yunnanensis seed oil and Cirsium japonicum fruit oil can improve the stability of the formulation, and the two have a synergistic effect in improving stability.

[0200] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0201] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0202] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A method for preparing an extract of Camellia yunnanensis, characterized in that, The preparation method of the Yunnan camellia flower extract includes the following steps: (1) Mix the Yunnan camellia flower raw material with water and enzymes, and perform enzymatic hydrolysis. The enzymes are pectinase and cellulase in a mass ratio of (1-3):

1. Then, the enzymes are inactivated to obtain an enzymatic hydrolysis solution. The enzymatic hydrolysis solution is subjected to ultrasonic treatment, and then solid-liquid separation is performed. The clear liquid is concentrated and dried to obtain crude extract of Yunnan camellia flower. (2) Mix the crude extract of Camellia yunnanensis with a two-phase extractant, stir at 30-50℃ for 30-60 min, let stand for layering and extract, take the upper phase after extraction treatment for concentration and drying to obtain the Camellia yunnanensis extract. The upper phase of the aqueous two-phase extractant is an aqueous solution containing 20-30 wt% ethanol, and the lower phase of the aqueous two-phase extractant is an aqueous solution containing 10-20% water-soluble salt, with the pH value adjusted to 5.0-7.0 using citric acid; the water-soluble salt is a combination of sodium citrate and ammonium sulfate in a mass ratio of (1-3):

1.

2. The method for preparing the Yunnan camellia flower extract according to claim 1, characterized in that, The amount of enzyme added is 0.1-1% of the enzymatic hydrolysis system.

3. The method for preparing the Yunnan camellia flower extract according to claim 1, characterized in that, The mass ratio of the Yunnan camellia flower raw material to water is 1:(5-20).

4. The method for preparing the Camellia yunnanensis flower extract according to claim 1, characterized in that, The pH value of the enzymatic hydrolysis system is 4-7, the temperature of the enzymatic hydrolysis treatment is 30-50℃, and the time of the enzymatic hydrolysis treatment is 30-60 min.

5. The method for preparing the Camellia yunnanensis flower extract according to claim 1, characterized in that, The ultrasonic treatment power is 100-400 W, the ultrasonic treatment time is 30-60 min, and the ultrasonic treatment temperature is 30-50℃.

6. The method for preparing the Yunnan camellia flower extract according to claim 1, characterized in that, The ratio of crude extract of Camellia yunnanensis to aqueous two-phase extractant is 1:(5-20) g / mL.

7. The method for preparing the Yunnan camellia flower extract according to claim 1, characterized in that, The concentration in steps (1) and (2) is carried out by vacuum concentration.

8. The method for preparing the Yunnan camellia flower extract according to claim 1, characterized in that, The drying process described in steps (1) and (2) is freeze-drying.

9. Camellia yunnanensis extract prepared by any one of claims 1-8.

10. A preparation containing an extract of Camellia yunnanensis, characterized in that, The raw materials for preparing the preparation include the following components: lecithin, phytosterols, Camellia yunnanensis seed oil, Prickly pear fruit oil, Camellia yunnanensis flower extract as described in claim 9, and cyclodextrin.

11. The formulation according to claim 10, characterized in that, The lecithin includes any one or a combination of at least two of soybean lecithin, egg yolk lecithin, hydrogenated lecithin, dipalmitoylphosphatidylcholine, or dioleoylphosphatidylcholine; The phytosterols include any one or a combination of at least two of β-sitosterol, stigmasterol, campesterol, or sitosterol. The cyclodextrin includes hydroxypropyl-β-cyclodextrin.

12. The formulation according to claim 10, characterized in that, The raw materials for preparing the preparation include the following components by mass fraction: 35-45% lecithin, 2-4% phytosterols, 2-4% Camellia yunnanensis seed oil, 1-4% Citrus aurantium fruit oil, 6-8% Camellia yunnanensis flower extract as described in claim 9, and 12-16% cyclodextrin.

13. The formulation according to claim 10, characterized in that, The raw materials for preparing the formulation also include a freeze-drying protectant selected from any one or a combination of at least two of trehalose, sucrose, mannose, mannitol, sorbitol, glycerol, polyvinylpyrrolidone, or polyethylene glycol.

14. The method for preparing a formulation containing Camellia yunnanensis flower extract according to any one of claims 10-13, characterized in that, The preparation method includes the following steps: (1) Dissolve the Yunnan camellia flower extract in a buffer solution and then mix it with cyclodextrin to obtain solution A; Lecithin, phytosterols, Camellia yunnanensis seed oil, and Prickly pear fruit oil were dissolved in anhydrous ethanol and then evaporated to obtain a lipid film. (2) Solution A is added to the lipid film for hydration treatment, and then homogenized to obtain the preparation containing the Yunnan camellia extract.

15. The preparation method according to claim 14, characterized in that, The pH of the buffer solution is 4-7.

16. The preparation method according to claim 14, characterized in that, The mixing and stirring are carried out at 20-40℃ for 2-6 hours.

17. The preparation method according to claim 14, characterized in that, The hydration treatment is carried out at 40-60℃ for 1-3 hours.

18. The preparation method according to claim 14, characterized in that, The homogenization process was performed in 1-5 cycles at 8000-10000 psi, each cycle lasting 2-5 minutes.

19. The preparation method according to claim 14, characterized in that, The homogenized product is then freeze-dried after homogenization.

20. The use of the Camellia yunnanensis extract of claim 9 or any of the preparations containing Camellia yunnanensis extract according to any one of claims 10-13 in the preparation of products with whitening effects.

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