Preparation method of camellia yunnanensis flower extract as well as product and application of camellia yunnanensis flower extract

Abstract

The invention relates to a preparation method of a camellia Yunnan flower extract and a product and application thereof, the preparation method comprises the following steps: mixing a camellia Yunnan flower raw material with water and enzyme, carrying out enzymolysis treatment, and then carrying out enzyme deactivation treatment; performing ultrasonic treatment, solid-liquid separation, clear liquid concentration and drying on the enzymolysis solution to obtain a crude flower extract; mixing the crude extract with an aqueous two-phase extraction agent for extraction, and concentrating and drying an upper phase to obtain the camellia yunnanensis flower extract. According to the extraction process, the extract with tyrosinase inhibitory activity can be obtained to the greatest extent at a relatively low temperature; a two-aqueous-phase extraction method is adopted to enrich multiple target active components, and it is ensured that the extracted product has more excellent tyrosinase inhibitory activity. The invention further develops a preparation containing the camellia Yunnan flower extract and having excellent stability, the camellia Yunnan flower extract is entrapped by cyclodextrin, the inclusion compound is loaded in a liposome structure, and the stability of the obtained preparation can be further improved by adding the camellia Yunnan seed oil and the prinsepia utilis royle oil.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of plant extraction, and relates to a preparation method of Camellia reticulata Lindl flower extract, a product thereof and application. BACKGROUND

[0002] The color of skin is mainly determined by the content and distribution of skin pigments, and melanin is the most important determinant. Melanocytes in the skin produce melanin, and melanin granules are transferred to keratinocytes through the dendritic processes of melanocytes. The melanin granules transferred to keratinocytes move up with the epidermal cells to the stratum corneum, thereby affecting the color of the skin or forming a color spot.

[0003] At present, general skin whitening agents mainly inhibit tyrosinase activity or block the synthesis pathway of tyrosine to generate melanin, thereby reducing the generation of melanin to achieve the effect of whitening. Such skin whitening agents mainly include arbutin, hydroquinone and derivatives, L-ascorbic acid and derivatives, kojic acid and derivatives, vitamin C ethyl ether and derivatives, nicotinamide, trisaminic acid, benzenediol and derivatives, and the like. With the increasing demand of consumers for safe and natural ingredients, high-efficiency and low-irritation natural tyrosinase inhibitors will become an important development direction of the cosmetic industry.

[0004] Camellia reticulata Lindl flowers are rich in active ingredients such as polyphenolic compounds and water-soluble pigments, which have tyrosinase inhibitory activity and can effectively block the synthesis of melanin in vivo and in vitro, and are some natural and efficient whitening active ingredients. However, how to extract Camellia reticulata Lindl flowers to obtain extracts with higher tyrosinase inhibitory activity is rarely reported in the prior art. Meanwhile, the color stability of Camellia reticulata Lindl flower extract is greatly affected by the structure of chemical components and environmental factors (such as light, heat, pH, and oxygen), which limits its application in cosmetic formulations to some extent. SUMMARY

[0005] In view of the deficiencies of the prior art, the present application aims to provide a preparation method of Camellia reticulata Lindl flower extract, a product thereof and application, and particularly relates to a preparation method of Camellia reticulata Lindl flower extract with tyrosinase inhibitory activity and whitening effect, a product thereof, a preparation method of a high-stability preparation containing the aforementioned Camellia reticulata Lindl flower extract, and application of the aforementioned Camellia reticulata Lindl flower extract or preparation in the preparation of tyrosinase inhibitors or products with whitening effect.

[0006] To achieve the purpose of the present application, the following technical solutions are adopted:

[0007] In a first aspect, the present application provides a preparation method of Camellia reticulata Lindl flower extract, which comprises 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 limitation of Camellia yunnanensis extract in cosmetic formulation due to its color instability, 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 lecithin mass fraction can be selected from 35%, 38%, 40%, 42%, 43%, 44%, 45%, etc.; the phytosterol mass fraction can be selected from 2%, 2.5%, 3%, 3.5%, 4%, etc.; the Yunnan camellia seed oil mass fraction can be selected from 2%, 2.5%, 3%, 3.5%, 4%, etc.; the prickly ash fruit oil mass fraction can be selected from 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, etc.; the Yunnan camellia flower extract mass fraction can be selected from 6%, 6.5%, 7%, 7.5%, 8%, etc.; the cyclodextrin mass fraction 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 this preparation method and 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 enzymes (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 reaction was carried out in a 96-well plate, and the order of sample addition 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 Camellia reticulata Lindl. flower extract, characterized by, The preparation method of the Camellia reticulata Lindl flower extract comprises the following steps: (1) mixing Camellia reticulata Lindl flower raw materials with water and enzymes, performing enzymatic hydrolysis treatment, then performing enzyme inactivation treatment to obtain an enzymatic hydrolysis solution; performing ultrasonic treatment on the enzymatic hydrolysis solution, then performing solid-liquid separation, concentrating and drying the clear liquid to obtain a Camellia reticulata Lindl flower crude extract; (2) mixing the Camellia reticulata Lindl flower crude extract with a two-water-phase extraction agent to perform extraction, taking the upper phase after the extraction treatment to perform concentration and drying to obtain the Camellia reticulata Lindl flower extract; The upper phase of the two-water-phase extraction agent is an aqueous solution containing ethanol, and the lower phase of the two-water-phase extraction agent is an aqueous solution containing a water-soluble salt.

2. The method of claim 1, wherein the Camellia reticulata extract is prepared by the steps of, The enzymes include pectinase and cellulase; the mass ratio of the pectinase to the cellulase is (1-3):1; The addition amount of the enzymes is 0.1-1% of the enzymatic hydrolysis system; The mass ratio of the Camellia reticulata Lindl flower raw materials to water is 1:(5-20); 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.

3. The method of claim 1, wherein the Camellia reticulata Lindl. flower extract is prepared by the steps of: The power of the ultrasonic treatment is 100-400 W, the time of the ultrasonic treatment is 30-60 min, and the temperature of the ultrasonic treatment is 30-50℃; The upper phase of the two-water-phase extraction agent is an aqueous solution containing 20-30 wt% ethanol, and the lower phase of the two-water-phase extraction agent is an aqueous solution containing 10-20% water-soluble salt; 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, and ammonium sulfate.

4. The method of claim 1, wherein the Camellia reticulata extract is prepared by the steps of, The water-soluble salt is a combination of sodium citrate and ammonium sulfate, and the mass ratio of the sodium citrate to the ammonium sulfate is (1-3):

1.

5. The method of claim 1, wherein the Camellia reticulata extract is prepared by the steps of: The solid-liquid ratio of the Camellia reticulata Lindl flower crude extract to the two-water-phase extraction agent is 1:(5-20) g / mL; The extraction treatment is stirred at 30-50℃ for 30-60 min, and then static layering is performed; The two-water-phase extraction agent system further adopts citric acid to adjust the pH value to 5.0-7.0; The concentration in steps (1) and (2) adopts a reduced-pressure concentration mode; The drying in steps (1) and (2) adopts a freeze-drying mode.

6. The Camellia reticulata Lindl flower extract prepared by the preparation method in any one of claims 1-5.

7. A preparation comprising an extract of Camellia reticulata Lindl. flowers, characterized in that, The preparation raw materials of the preparation include the following components: lecithin, phytosterol, Camellia reticulata seed oil, Prinsepia uniflora seed oil, the Camellia reticulata Lindl flower extract in claim 6, and cyclodextrin.

8. The formulation of claim 6, wherein, The lecithin includes any one or a combination of at least two of soybean lecithin, egg yolk lecithin, hydrogenated lecithin, dipalmitoyl phosphatidylcholine, or dioleoyl phosphatidylcholine; The phytosterol includes any one or a combination of at least two of β-sitosterol, stigmasterol, campesterol, or sitostanol; The cyclodextrin includes hydroxypropyl-β-cyclodextrin; The preparation raw materials of the preparation include the following components in a mass fraction: lecithin 35-45%, phytosterol 2-4%, Camellia reticulata seed oil 2-4%, Prinsepia uniflora seed oil 1-4%, the Camellia reticulata Lindl flower extract in claim 6 6-8%, and cyclodextrin 12-16%; The raw materials for preparing the preparation also include optional lyophilization protectants selected from any one or a combination of at least two of trehalose, sucrose, mannose, mannitol, sorbitol, glycerol, polyvinylpyrrolidone or polyethylene glycol.

9. The method for preparing a formulation containing Camellia yunnanensis flower extract according to claim 7 or 8, characterized in that, The preparation method comprises the following steps: (1) dissolving Camellia pitardii flower extract in a buffer solution and then mixing and stirring with cyclodextrin to obtain solution A; dissolving lecithin, phytosterol, Camellia pitardii seed oil and Prinsepia uniflora oil in an ethanol solution for evaporation to obtain a lipid film; (2) adding solution A to the lipid film for hydration treatment and then homogenization treatment to obtain the preparation containing Camellia pitardii flower extract; the pH of the buffer solution is 4-7; the mixing and stirring is performed at 20-40°C for 2-6 h; the hydration treatment is performed at 40-60°C for 1-3 h; the homogenization treatment is performed at 8000-10000 psi for 1-5 cycles, each cycle being 2-5 min; the homogenization treatment is further optionally followed by freeze-drying treatment of the homogenization product.

10. Use of Camellia pitardii flower extract of claim 6 or the preparation containing Camellia pitardii flower extract of claim 7 or 8 in the preparation of a product having a whitening effect or in the preparation of a tyrosinase inhibitor.

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