Compound containing wild jujube leaves and ethyl acetate as well as preparation and application of compound

Abstract

The invention relates to the technical field of cosmetics, in particular to a compound containing wild jujube leaves and ethyl acetate as well as preparation and application of the compound. The compound containing the wild jujube leaf ethyl acetate phase comprises the following components in percentage by mass: celery seed oil and the wild jujube leaf ethyl acetate phase according to a mass ratio of 1: (0.1-15). According to the invention, the celery seed oil and the wild jujube leaf ethyl acetate are compounded to achieve the effects of synergistically inhibiting bacteria, resisting oxidation, whitening and the like, the celery seed oil has a relatively good transdermal absorption promoting effect and can promote the transdermal absorption amount of the wild jujube leaf ethyl acetate phase, and the celery seed oil and the wild jujube leaf ethyl acetate are compounded and added into a gel mask or cream, so that the skin whitening effect is achieved. And the prepared gel mask and cream (containing the compound containing the wild jujube leaves and the ethyl acetate) have the effects of synergistically inhibiting bacteria, resisting oxidation, whitening and the like, and also have the effect of promoting permeation.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetics, and in particular to a compound containing ethyl acetate phase of wild jujube leaves, and the preparation and application thereof. Background Art

[0002] Natural plants have always been an important resource for mankind. In recent years, research on natural plants has made significant progress.

[0003] Celery seed oil is a yellowish oil extracted from celery seeds. It has the effects of removing edema, promoting skin smoothness, increasing blood flow, lowering blood pressure, and improving brain microcirculation. It has been widely used in the fields of medicine, food, and cosmetics. There are many active ingredients and pharmacological effects in the leaves of the Chinese jujube, mainly including flavonoids, polyphenols, polysaccharides, saponins, alkaloids, triterpenes, etc. It has antioxidant, anti-aging, antibacterial, anti-inflammatory, and protective effects on the nervous system.

[0004] However, there is no research on the combination of celery seed oil and jujube leaf extract. Summary of the invention

[0005] In order to solve the above problems, the purpose of the present invention is to provide a compound containing ethyl acetate phase of sour jujube leaves and its preparation and application.

[0006] The invention provides a composite material containing ethyl acetate phase of jujube leaves, which has synergistic antibacterial, anti-oxidation and whitening effects, especially high sensitivity to antibacterial activity. Compared with its single species, the composite material significantly improves the penetration of functional substances and prolongs the shelf life of the product.

[0007] The invention adds a compound containing ethyl acetate phase of wild jujube leaves to a gel mask and a cream, determines the biological activities of the compound such as synergistic antibacterial, anti-oxidation, whitening, and penetration promotion, prepares the compound containing ethyl acetate phase of wild jujube leaves and a gel mask and a cream with multiple functional effects such as antibacterial, and performs physical and chemical properties and sensory evaluation on the compound with reference to current industry standards; the obtained gel mask and cream containing ethyl acetate phase of wild jujube leaves have the advantages of high safety, obvious synergistic effect, prolonged shelf life, and the like.

[0008] The purpose of the present invention can be achieved by the following technical solutions:

[0009] The first object of the present invention is to provide a composite material containing ethyl acetate phase of jujube leaves, comprising the following components in percentage by mass:

[0010] Celery seed oil: jujube leaf ethyl acetate phase = 1:0.1~15.

[0011] In one embodiment of the present invention, the celery seed oil is prepared by the following method:

[0012] The celery seeds are crushed and soaked in a sodium chloride aqueous solution, and then heated in an oil bath and post-treated to obtain celery seed oil;

[0013] Wherein, the mass fraction of sodium chloride in the sodium chloride aqueous solution is 3%;

[0014] The material-liquid ratio of celery seeds to sodium chloride aqueous solution is 1:25-30;

[0015] During the soaking process, the temperature is room temperature and the time is 3 to 4 hours;

[0016] During the oil bath heating process, the temperature is 130-140°C and the time is 4-5h;

[0017] The post-treatment was to remove water using anhydrous sodium sulfate.

[0018] In one embodiment of the present invention, the ethyl acetate phase of sour jujube leaves is prepared by the following method:

[0019] The wild jujube leaves are crushed and placed in an ethanol solution, heated under reflux, and then post-processed to obtain a wild jujube leaf alcohol extract, and then the wild jujube leaf alcohol extract is dissolved in distilled water, and then extracted with ethyl acetate, and the upper layer solution is evaporated to remove the solvent to obtain a wild jujube leaf ethyl acetate phase;

[0020] Wherein, the mass fraction of ethanol in the ethanol solution is 70-80%;

[0021] The sour jujube leaves are dried sour jujube leaves;

[0022] The solid-liquid ratio of the sour jujube leaves to the ethanol solution is 1:10-15;

[0023] During the reflux process, the temperature is 85-95°C and the time is 2-3h;

[0024] The post-treatment is to filter out the residue and then perform rotary evaporation concentration treatment;

[0025] The usage ratio of distilled water to the alcohol extract of Ziziphus jujuba leaves is 40-60 mL:1 g, and the usage of ethyl acetate (ethyl acetate is extracted for 2-4 times) is the same as that of distilled water.

[0026] In one embodiment of the present invention, the following components are included in percentage by weight:

[0027] Celery seed oil: jujube leaf ethyl acetate phase = 1:0.2~13.

[0028] In one embodiment of the present invention, the following components are included in percentage by weight:

[0029] Celery seed oil: ethyl acetate phase of jujube leaves = 1:5;

[0030] Celery seed oil: ethyl acetate phase of jujube leaves = 1:9;

[0031] Celery seed oil: jujube leaf ethyl acetate phase = 1:11.

[0032] In one embodiment of the present invention, the following components are included in percentage by weight:

[0033] Celery seed oil: ethyl acetate phase of jujube leaves = 1:11 (under this ratio, the synergistic antibacterial, antioxidant, and tyrosinase inhibition effects are the best).

[0034] The second object of the present invention is to provide a method for preparing a composite material containing ethyl acetate phase of wild jujube leaves, comprising the following steps:

[0035] The celery seed oil and the ethyl acetate phase of the wild jujube leaves are mixed evenly to obtain a composite material containing the ethyl acetate phase of the wild jujube leaves.

[0036] The third object of the present invention is to provide a use of a complex containing ethyl acetate phase of wild jujube leaves in the preparation of a facial mask and / or cream with synergistic antibacterial, anti-oxidation, whitening, and penetration-promoting functions.

[0037] The fourth object of the present invention is to provide a facial mask, comprising a compound containing ethyl acetate phase of wild jujube leaves.

[0038] The fifth object of the present invention is to provide a cream comprising a compound containing ethyl acetate of jujube leaves.

[0039] Celery seed oil has the effects of anti-oxidation, anti-tumor, lowering blood lipids, lowering blood pressure, antibacterial, anti-aging, removing edema, and promoting skin smoothness; the wild jujube leaf has the effects of anti-oxidation, anti-aging, antibacterial, anti-inflammatory, and protecting the nervous system. The synergistic effect of celery seed oil and wild jujube leaf ethyl acetate in a certain proportion range can achieve the effects of 1+1>2, such as antibacterial, anti-oxidation, and whitening. The resulting gel mask and cream have better antibacterial and anti-oxidation functions, and at the same time extend the shelf life of the product.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] (1) In the present invention, celery seed oil and ethyl acetate of jujube leaves are compounded, and the optimal compounding ratio of the compound is determined to be 1:11; the compound containing ethyl acetate of jujube leaves is applied to gel masks and creams, which has certain synergistic antioxidant, whitening, antibacterial and other effects, wherein the antibacterial effect reaches high sensitivity. The product also has the properties of promoting transdermal absorption and having a long shelf life.

[0042] (2) The cream and gel mask of the present invention have natural plant extract as the main active ingredients, which are highly safe and mild to the skin without irritation. Celery seed oil has the effects of anti-oxidation, anti-tumor, lowering blood lipids, lowering blood pressure, anti-bacterial, anti-aging, removing edema, and promoting skin smoothness. Jujube leaves have the effects of anti-oxidation, anti-aging, anti-bacterial, anti-inflammatory, and protecting the nervous system.

[0043] The present invention adopts the ethyl acetate phase compound containing jujube leaves as the effective substance. It is not just a simple combination of the two substances, but through reasonable compatibility, the compound achieves better results in biological activities such as synergistic antibacterial, anti-oxidation, whitening, and penetration promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 The antibacterial activity results of the composites containing ethyl acetate phase of jujube leaves at different mass ratios; (A) Staphylococcus aureus; (B) Escherichia coli;

[0045] Figure 2 The results of the scavenging rate of ABTS free radicals by the ethyl acetate phase compound containing jujube leaves at different mass ratios;

[0046] Figure 3 The results of the scavenging rate of DPPH free radicals by the ethyl acetate phase compound containing jujube leaves at different mass ratios;

[0047] Figure 4 The results of inhibition of tyrosinase by ethyl acetate phase compound containing jujube leaves in different mass ratios;

[0048] Figure 5 This is the result of the in vitro skin cumulative penetration of celery seed oil and ethyl acetate phase of jujube leaves;

[0049] Figure 6 This is a schematic diagram of the appearance of a blank matrix mask;

[0050] Figure 7 Schematic diagram of the appearance of the cream; (A) blank matrix cream; (B) cream containing celery seed oil and ethyl acetate of jujube leaves. DETAILED DESCRIPTION

[0051] The present invention provides a composite material containing ethyl acetate phase of wild jujube leaves, comprising the following components in the following mass percentages:

[0052] Celery seed oil: jujube leaf ethyl acetate phase = 1:0.1~15.

[0053] In one embodiment of the present invention, the celery seed oil is prepared by the following method:

[0054] The celery seeds are crushed and soaked in a sodium chloride aqueous solution, and then heated in an oil bath and post-treated to obtain celery seed oil;

[0055] Wherein, the mass fraction of sodium chloride in the sodium chloride aqueous solution is 3%;

[0056] The material-liquid ratio of celery seeds to sodium chloride aqueous solution is 1:25-30;

[0057] During the soaking process, the temperature is room temperature and the time is 3 to 4 hours;

[0058] During the oil bath heating process, the temperature is 130-140°C and the time is 4-5h;

[0059] The post-treatment was to remove water using anhydrous sodium sulfate.

[0060] In one embodiment of the present invention, the ethyl acetate phase of sour jujube leaves is prepared by the following method:

[0061] The wild jujube leaves are crushed and placed in an ethanol solution, heated under reflux, and then post-processed to obtain a wild jujube leaf alcohol extract, and then the wild jujube leaf alcohol extract is dissolved in distilled water, and then extracted with ethyl acetate, and the upper layer solution is evaporated to remove the solvent to obtain a wild jujube leaf ethyl acetate phase;

[0062] Wherein, the mass fraction of ethanol in the ethanol solution is 70-80%;

[0063] The sour jujube leaves are dried sour jujube leaves;

[0064] The solid-liquid ratio of the sour jujube leaves to the ethanol solution is 1:10-15;

[0065] During the reflux process, the temperature is 85-95°C and the time is 2-3h;

[0066] The post-treatment is to filter out the residue and then perform rotary evaporation concentration treatment;

[0067] The usage ratio of distilled water to the alcohol extract of Ziziphus jujuba leaves is 40-60 mL:1 g, and the usage of ethyl acetate (ethyl acetate is extracted for 2-4 times) is the same as that of distilled water.

[0068] In one embodiment of the present invention, the following components are included in percentage by weight:

[0069] Celery seed oil: jujube leaf ethyl acetate phase = 1:0.2~13.

[0070] In one embodiment of the present invention, the following components are included in percentage by weight:

[0071] Celery seed oil: ethyl acetate phase of jujube leaves = 1:5;

[0072] Celery seed oil: ethyl acetate phase of jujube leaves = 1:9;

[0073] Celery seed oil: jujube leaf ethyl acetate phase = 1:11.

[0074] In one embodiment of the present invention, the following components are included in percentage by weight:

[0075] Celery seed oil: ethyl acetate phase of jujube leaves = 1:11 (under this ratio, the synergistic antibacterial, antioxidant, and tyrosinase inhibition effects are the best).

[0076] The present invention provides a method for preparing a composite material containing ethyl acetate phase of wild jujube leaves, comprising the following steps:

[0077] The celery seed oil and the ethyl acetate phase of the wild jujube leaves are mixed evenly to obtain a composite material containing the ethyl acetate phase of the wild jujube leaves.

[0078] The present invention provides an application of a compound containing ethyl acetate phase of wild jujube leaves in preparing a facial mask and / or cream with synergistic antibacterial, anti-oxidation, whitening and penetration-promoting functions.

[0079] The invention provides a facial mask, comprising a compound containing ethyl acetate phase of wild jujube leaves.

[0080] The invention provides a cream comprising a compound containing ethyl acetate phase of wild jujube leaves.

[0081] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0082] In the following examples, unless otherwise specified, all reagents used are commercially available reagents, and all detection means and methods used are conventional detection means and methods in the art.

[0083] Example 1

[0084] The present embodiment provides a method for extracting celery seed oil (steam distillation method), comprising the following steps:

[0085] Celery seeds (crushed celery seeds) were mixed with distilled water containing sodium chloride (3% by mass) at a solid-liquid ratio of 1:30, soaked at room temperature for 3 hours, heated and extracted at 130°C for 5 hours, water was absorbed with anhydrous sodium sulfate, weighed, celery seed oil was obtained, and stored in a refrigerator for later use.

[0086] Example 2

[0087] This embodiment provides a method for extracting ethyl acetate phase from wild jujube leaves, comprising the following steps:

[0088] The dried wild jujube leaves were crushed, 130 g was weighed for standby use, and added into a round-bottom flask filled with 70% ethanol solvent at a solid-liquid ratio of 1:10 (M / V), heated and refluxed at 90°C for 2 hours, filtered while hot, and the residue was discarded. After rotary evaporation concentration at 80°C for 4 hours (it can be adjusted according to actual conditions, and rotary evaporation concentration at 75-85°C for 4-5 hours is acceptable), an alcohol extract of wild jujube leaves was obtained;

[0089] Dissolve 3 g of jujube leaf alcohol extract in 150 mL of distilled water, then extract with 150 mL of ethyl acetate three times, combine the upper solutions, and concentrate by rotary evaporation at 30°C for 1 h to remove the solvent to obtain the ethyl acetate phase of jujube leaves, which is refrigerated for later use.

[0090] Example 3

[0091] This embodiment provides a method for preparing a composite material containing ethyl acetate phase of wild jujube leaves, comprising the following steps:

[0092] The celery seed oil prepared in Example 1 and the ethyl acetate phase of wild jujube leaves prepared in Example 2 are mixed to obtain a composite containing ethyl acetate phase of wild jujube leaves;

[0093] Among them, the mass ratios of celery seed oil and ethyl acetate phase of jujube leaves were 9:1, 5:1, 1:1, 1:5, 1:9, 1:11, 1:13 and 1:15, respectively.

[0094] Example 4

[0095] This example provides the antibacterial activity determination of the ethyl acetate phase compound containing the wild jujube leaves (prepared in Example 3).

[0096] Using the filter paper method, accurately draw 100 μL of bacterial suspension (1×10 6 CFU / mL concentration of Staphylococcus aureus and 1×10 6coli with a CFU / mL concentration), added to a solid culture medium (nutrient agar), spread evenly, used sterilized tweezers to pick up a round filter paper (6 mm in diameter, sterilized) to which 15 μL of sample solution had been added (the sample solution was a mixture of celery seed oil, ethyl acetate phase of wild jujube leaves, or a compound containing ethyl acetate phase of wild jujube leaves and a solvent, wherein the concentrations of celery seed oil, ethyl acetate phase of wild jujube leaves, and the compound containing ethyl acetate phase of wild jujube leaves were all 6.25 mg / mL, and the solvents were all DMSO), placed it on a culture dish, and cultured in a constant temperature incubator at 37°C for 24 h (the whole process was under sterile conditions), took it out, and measured the diameter of the inhibition zone with a vernier caliper (using the cross method), wherein the blank control was DMSO, the addition amount was 15 μL, and the average value was taken for 3 parallel measurements. The antibacterial effect of the samples was compared by comparing the size of the inhibition zone diameter. The judgment criteria were: inhibition zone diameter > 20 mm was extremely sensitive, 15 < inhibition zone diameter ≤ 20 mm was highly sensitive, 9 < inhibition zone diameter ≤ 15 was moderately sensitive, 7 < inhibition zone diameter ≤ 9 mm was lowly sensitive, and no inhibition zone indicated insensitivity.

[0097] The antibacterial activity results of the mixtures of celery seed oil and ethyl acetate of jujube leaves at different mass ratios are shown in Table 1 and Figure 1 shown.

[0098] Table 1 Inhibition zone results of different mass ratios of the mixtures against two bacteria

[0099]

[0100]

[0101] pass Figure 1 It can be found from Table 1 that the antibacterial activity varies with the different compounding ratios. For Staphylococcus aureus, when the compounding ratio of celery seed oil and ethyl acetate phase of jujube leaves is 9:1 and 5:1, the size of the inhibition zone is less than 9 mm and smaller than the size of the inhibition zone of the ethyl acetate phase of jujube leaves alone, which is low sensitivity; as the proportion of ethyl acetate phase of jujube leaves increases, the size of the inhibition zone increases, and the inhibition zone reaches the maximum at the compounding ratio of 1:11, which is 17.5 mm, which is high sensitivity. As the proportion of ethyl acetate phase of jujube leaves continues to increase, it is found that the inhibition zone begins to decrease, and it is concluded that the compounding ratio (the mass ratio of celery seed oil to ethyl acetate phase of jujube leaves, the same below) is synergistic or increasing in the range of 1:1 to 1:15; for Escherichia coli, except for the compounding ratios of 9:1 and 5:1, the range from 1:1 to 1:15 is synergistic or increasing, and the best antibacterial effect is achieved at 1:11, with the inhibition zone reaching 12.5 mm, which is medium sensitivity. Combining celery seed oil with ethyl acetate from jujube leaves, the antibacterial activity of the two bacteria was studied at different mass ratios, and it was concluded that the range of the ratio for the synergistic effect was 1:1 to 1:15.

[0102] Example 5

[0103] This example provides the determination of the antioxidant activity (ABTS free radical and DPPH free radical) of the composite containing ethyl acetate phase of sour jujube leaves (prepared in Example 3).

[0104] Mix 1 mL of sample solution (the sample solution is a mixture of celery seed oil, ethyl acetate phase of jujube leaves, or a mixture containing ethyl acetate phase of jujube leaves and a solvent, wherein the concentrations of celery seed oil, ethyl acetate phase of jujube leaves, and the mixture containing ethyl acetate phase of jujube leaves are all 0.01 mg / mL, and the solvent is anhydrous ethanol) with 3 mL of ABTS working solution, place it at room temperature away from light for 10 minutes, and measure the absorbance at 734 nm. The blank control is anhydrous ethanol. Test three times in parallel. Calculate the sample's response to ABTS according to the formula + Clearance rate:

[0105]

[0106] In the formula, A i : 1mL sample solution and 3mL ABTS + The absorbance value of the solution; A j A is the absorbance value of 1 mL sample solution and 3 mL anhydrous ethanol; c 1mL of anhydrous ethanol and 3mL of ABTS + The absorbance value of the solution.

[0107] The synergistic coefficient (SE) is the ratio between the actual clearance rate (or inhibition rate, i.e., ESC) and the theoretical clearance rate (or inhibition rate, i.e., TSC) of the compound, i.e., SE = ESC / TSC.

[0108] The calculation formula of TSC is as follows:

[0109] TSC(%)=((ESC1+ESC2)-(ESC1×ESC2)) / 100

[0110] Where: ESC is the actual clearance rate or actual inhibition rate of the compound, ESC1 is the actual clearance rate or actual inhibition rate of celery seed oil / %, and ESC2 is the actual clearance rate or actual inhibition rate of the ethyl acetate phase of jujube leaves / %;

[0111] When SE>1, it means that the compound has a synergistic effect; when SE=1, the compound shows an additive effect; when SE<1, it is an antagonistic effect.

[0112] The results of ABTS free radical scavenging ability test are as follows Figure 2 As shown, through Figure 2It can be found that the SE value of the complex of celery seed oil and ethyl acetate of wild jujube leaves ranges from 0.51 to 1.03, and the complex ratio ranges from 1:5 to 1:9, which belongs to synergistic effect. When the ratio is 1:11, SE=0.99, which can also be judged as an additive effect.

[0113] An equal volume of sample solution (the sample solution is a mixture of celery seed oil, ethyl acetate phase of wild jujube leaves or a mixture containing ethyl acetate phase of wild jujube leaves and a solvent, wherein the concentrations of celery seed oil, ethyl acetate phase of wild jujube leaves and the mixture containing ethyl acetate phase of wild jujube leaves are all 0.01 mg / mL, and the solvent is all anhydrous ethanol) was uniformly mixed with an anhydrous ethanol solution containing 0.16 mmol / L DPPH (hereinafter referred to as "DPPH solution"), placed at room temperature away from light for 40 min, and the absorbance was measured at 517 nm. The blank control was anhydrous ethanol.

[0114] Calculate the DPPH of the sample according to the following formula + Clearance rate:

[0115]

[0116] In the formula, A i : absorbance value of 2mL sample solution and 2mL DPPH solution; A j : absorbance value of 2mL sample solution and 2mL anhydrous ethanol; A c : Absorbance value of 2mL anhydrous ethanol and 2mL DPPH solution.

[0117] DPPH free radical scavenging ability test results are as follows Figure 3 As shown, through Figure 3 It can be found that the SE value of celery seed oil and jujube leaf ethyl acetate phase ranges from 0.55 to 1.25, and the compound ratio ranges from 1:5 to 1:13, which is a synergistic effect. The synergistic effect is best when the ratio is 1:5, and the actual clearance rate is 25%, SE = 1.25.

[0118] Example 6

[0119] This example provides a determination of the inhibition of tyrosinase activity of a compound containing ethyl acetate phase of wild jujube leaves (prepared in Example 3).

[0120] The experimental group reaction solution includes 1.5mL of 0.05mM PBS buffer solution (pH 6.8), 0.6mL of sample solution (the sample solution is a mixture of celery seed oil, ethyl acetate phase of jujube leaves or a mixture of ethyl acetate phase containing jujube leaves and a solvent, wherein the concentrations of celery seed oil, ethyl acetate phase of jujube leaves and the mixture of ethyl acetate phase containing jujube leaves are all 0.2mg / mL, and the solvents are all deionized water), 0.3mL of tyrosinase solution with a concentration of 0.3mg / mL, stand at room temperature for 10min, add 0.6mL of L-dopa (L-DOPA, 0.01M) solution, mix and react for 10min, and detect the absorbance value at a wavelength of 475nm. The control group is replaced with an equal volume of deionized water, and the parallel determination is repeated 3 times to take the average value. The tyrosinase inhibition rate (%) is calculated according to the following formula:

[0121]

[0122] Table 2 Composition of reaction solution

[0123]

[0124] The results of tyrosinase inhibition test were as follows Figure 4 As shown by Figure 4 It can be seen that celery seed oil and ethyl acetate phase of jujube leaves have a synergistic effect within a certain range of compounding ratios, with SE values ​​ranging from 0.81 to 1.54. In the compounding ratio range of 1:1 to 1:15, the inhibitory effect on tyrosinase is synergistic, and the synergistic effect is best at a ratio of 1:5, with an actual inhibition rate of 48.98%.

[0125] According to the three groups of experimental measurements, the compound ratios with synergistic antioxidant and enzyme inhibition activities were obtained, as shown in Table 3.

[0126] Table 3 Compound ratios with optimal antioxidant and enzyme inhibition activities

[0127]

[0128] As shown in Table 3, the ratios of 1:5, 1:9 and 1:11 have antibacterial, antioxidant and tyrosinase inhibition activities at the same time, and the ratio of 1:11 is the best for antibacterial activity, and the antioxidant and tyrosinase inhibition activities also have good synergistic and additive effects at this ratio. Therefore, the compounding ratio of 1:11 was selected for the next experiment. (The compounding materials mentioned later are all in the mass ratio of celery seed oil: ethyl acetate phase of jujube leaves = 1:11)

[0129] Example 7

[0130] This example provides the determination of MIC, MBC and FIC values ​​of a composite containing ethyl acetate phase of wild jujube leaves (the mass ratio of celery seed oil to ethyl acetate phase of wild jujube leaves is 1:11).

[0131] (1) Determination of MIC and MBC

[0132] The micro-broth dilution method was used. Each column of the 96-well plate was marked as AL, each row was marked as 1-8, and every three columns were three parallel groups of the same sample. First, 100 μL of sterilized nutrient broth was added to each well, and then 100 μL of sample solution with a concentration of 100 mg / mL was added to the first row (the sample solution was a mixture of celery seed oil, ethyl acetate phase of jujube leaves, or a mixture of ethyl acetate phase of jujube leaves and a solvent, wherein the concentrations of celery seed oil, ethyl acetate phase of jujube leaves, and ethyl acetate phase of jujube leaves were all 6.25 mg / mL, and the solvents were all DMSO), mixed evenly, 100 μL was drawn to the second row, mixed evenly, and the same steps were repeated to rows 3-8, 100 μL of the solution in row 8 was drawn and discarded, and finally 100 μL of 1×10 6 CFU / mL Staphylococcus aureus suspension, mix well, culture in a 37℃ constant temperature incubator for 24h, add 15μL of 5mg / mL MTT solution to each well, and take the lowest concentration of sterile growth as the minimum inhibitory concentration (MIC) of the sample according to the color change of each well (if bacteria grow, the color of the well turns black, otherwise there is no change). Then take 100μL of the sterile growth sample solution on the solid culture medium (nutrient agar), spread evenly, and the lowest concentration with a colony count of ≤5 after 24h of constant temperature culture at 37℃ is defined as the minimum bactericidal concentration (MBC) of the sample. Among them, sterile nutrient broth without sample solution and Staphylococcus aureus suspension were used as controls.

[0133] (2) FIC determination

[0134] The combined antibacterial activity (FIC) of the compound was tested by the chessboard dilution method. Celery seed oil was recorded as A, and the ethyl acetate phase of jujube leaves was recorded as B. The single sample to be tested was diluted with sterilized nutrient broth to concentrations of MIC, 1 / 2MIC, 1 / 4MIC and 1 / 8MIC. The samples of the above concentrations were compounded in each test tube as shown in the chessboard below for later use; then 100 μL of the solution was drawn and added to the 96-well plate in the same order, and then 100 μL of a concentration of 1×10 6 CFU / mL of bacterial suspension so that its distribution in a 96-well plate is as shown in the checkerboard below.

[0135] Table 4 Schematic diagram of chessboard distribution

[0136]

[0137]

[0138] Sterile nutrient broth was used as blank control. After mixing evenly, the antibacterial activity of the compound was determined according to the MIC method. FIC was used as the basis for determining the combined antibacterial effect, and the calculation formula of FIC was:

[0139]

[0140] Where MIC A Combination refers to the concentration of substance A in the compound, MIC A Single use refers to the MIC value of a single substance A; MIC B Combination refers to the concentration of substance B in the compound, MIC B Single use refers to the MIC value of single substance B.

[0141] The criteria for determining the FIC value are: when 0<FIC≤0.5, the combined antibacterial efficacy after compounding is synergistic; when 0.5<FIC≤1, it is additive; when 1<FIC≤2, it is irrelevant; when FIC>2, it is antagonistic.

[0142] The results are shown in Tables 5 and 6.

[0143] Table 5 MIC and MBC values ​​of single substances and compound substances against two bacteria

[0144]

[0145] Table 6 Combined antibacterial effect of the compound

[0146]

[0147]

[0148] As shown in Table 5, the MIC value of the compound against Staphylococcus aureus is 0.39 mg / mL, and the MBC value is 0.78 mg / mL. The MIC value against Escherichia coli is 1.56 mg / mL, and the MBC value is 6.25 mg / mL, which are all stronger than the antibacterial activity of the single substance. This once again shows that the two monomer substances (celery seed oil and ethyl acetate phase of jujube leaves) have a synergistic effect after compounding, and the synergistic effect against Escherichia coli is more obvious than that against Staphylococcus aureus.

[0149] As can be seen from Table 6, the combination of celery seed oil and ethyl acetate of jujube leaves has a good combined antibacterial effect. For Staphylococcus aureus, the MIC value of celery seed oil alone was 12.50 mg / mL, which was reduced to 1.56 mg / mL in the combination, and the ethyl acetate phase of jujube leaves also decreased from 0.78 mg / mL to 0.10 mg / mL, and the FIC value was 0.25 (<0.5), showing a synergistic effect; for Escherichia coli, the MIC values ​​of these two substances in the combination were lower than the MIC values ​​in the single substance, and the FIC value was 0.375 (<0.5), showing a synergistic effect.

[0150] Example 8

[0151] This example provides a skin permeability measurement of a compound containing ethyl acetate phase of jujube leaves (the mass ratio of celery seed oil to ethyl acetate phase of jujube leaves is 1:11).

[0152] The improved Franz diffusion cell method was used to detect the percutaneous absorption promoting effect of each sample. Fresh pig skin was used to simulate human skin, subcutaneous fat was removed, and the dermis was retained. It was washed with 0.01 mol / L PBS (pH=7.4) for use. PBS was used as the receiving solution, and the sample solution with a mass concentration of 1 mg / mL (the sample solution was a mixture of celery seed oil, ethyl acetate phase of jujube leaves, or a mixture of ethyl acetate phase of jujube leaves and a solvent, and the solvent was deionized water) was used as the diffusion solution. The pig skin was sandwiched between the diffusion cell and the receiving cell. The experimental temperature was 32°C and the magnetic speed was 550r / min. The receiving solution taken after 12 hours was tested to determine the sample mass concentration, and the cumulative permeation per unit area was calculated according to the following formula:

[0153]

[0154] Where: Q is the cumulative permeability per unit area, mg / cm 2 ; ρ is the sample mass concentration at 12h, mg / mL; V is the volume of the receiving pool, 6.0mL; A is the effective skin area for penetration, 2.26cm 2 .

[0155] Depend on Figure 5 It can be seen that when acting as a single substance, the in vitro cumulative skin permeation of celery seed oil is much higher than that of the ethyl acetate phase of jujube leaves; when acting on a compound containing the ethyl acetate phase of jujube leaves, the permeation of the ethyl acetate phase of jujube leaves is significantly increased compared with the single substance, increasing by nearly 3 times, and there is not much change in the ratio of celery seed oil to the single substance. The results show that celery seed oil in the compound has the effect of promoting the penetration of functional substances.

[0156] Examples 9-12

[0157] Embodiments 9-12 provide a gel mask comprising a compound containing ethyl acetate phase of jujube leaves.

[0158] The total amount of each formula is 100g, calculated by mass percentage.

[0159] (S1) 0.4 g konjac gum and 0.6 g pectin are completely dissolved in 80° C. deionized water and set aside; 0.4 g glycerol is then added, heated in a water bath with stirring at a stirring rate of 300 r / min, and finally 0.4 g sodium hyaluronate is added, and deionized water is added to make the total mass 100 g, and after mixing, a colloid liquid with uniform material distribution and suitable viscosity is obtained; the gel matrix mask (such as Figure 6 As shown), the surface of the gel matrix mask is smooth, uniform and transparent, and has good stretchability and hardness; it is recorded as Example 9.

[0160] Using a facial mask matrix process, 80°C deionized water, 0.4g konjac gum, and 0.6g pectin are completely dissolved and set aside; then 0.4g glycerol is added, heated in a water bath and stirred at a stirring rate of 300r / min, and finally 0.4g sodium hyaluronate is added, and deionized water is added to make the total mass 99g, and after mixing, a colloid liquid with uniform material distribution and suitable viscosity is obtained;

[0161] 1 g of celery seed oil, ethyl acetate phase of wild jujube leaves, and a mixture of celery seed oil and ethyl acetate phase of wild jujube leaves (celery seed oil: ethyl acetate phase of wild jujube leaves = 1:11) were added to the obtained colloidal liquid to make a total mass of 100 g, and stirred evenly to obtain a gel solution;

[0162] (S2) Place a gel solution of celery seed oil (prepared in Example 1), ethyl acetate phase of jujube leaves (prepared in Example 2), and a composite containing ethyl acetate phase of jujube leaves (celery seed oil: ethyl acetate phase of jujube leaves = 1:11) in a mask mold, cool to room temperature, and press into a film to obtain a gel mask. Among them, the gel mask containing celery seed oil is recorded as Example 10, the gel mask containing ethyl acetate phase of jujube leaves is recorded as Example 11, and the gel mask containing the composite of ethyl acetate phase of jujube leaves is recorded as Example 12.

[0163] Table 7 Effects of different extract samples on gel mask

[0164]

[0165] Note: The pH (25°C) of gel masks in the current industry standard is in the range of 3.5 to 8.5 and meets the requirements.

[0166] It can be seen from Table 7 that the plant extracts (celery seed oil, ethyl acetate phase of jujube leaves) have no effect on the appearance of the gel mask matrix. In terms of aroma, Examples 10 and 12 containing celery seed oil have a faint celery fragrance, but meet the specified fragrance type and are gentle and natural; in terms of pH, different additive formulas have different pH values, but they are all within the industry standard range and will not cause irritation to the skin; at the same time, the cold resistance, heat resistance and centrifugation experiments of the gel masks with different additives were strictly tested, and it was found that no stratification occurred, indicating that the texture and performance of the gel mask after adding the sample can remain stable under different temperature conditions, and no stratification or precipitation will occur during transportation and use; in terms of shelf life, the shelf life of the formulas containing additives is extended, and the formula with the compound containing ethyl acetate phase of jujube leaves has the longest shelf life, reaching 14 days, which is more than twice that of the blank matrix, which may be attributed to the good antibacterial effect of the compound.

[0167] Examples 13-16

[0168] Embodiments 13-16 provide a cream containing a compound of ethyl acetate phase of Ziziphus jujuba leaves.

[0169] A cream containing a compound of ethyl acetate phase of jujube leaves mainly comprises the following components in percentage by weight: 3% APG, 4% stearic acid, 1% castor oil, 0.5% monoglyceride, 1% dimethicone, 4% polydimethylsiloxane, 8% coconut oil, 3% 1,3-propylene glycol, 5% glycerol, 0.02% hyaluronic acid, 0.2% hydroxyethyl cellulose, and 0.4% xanthan gum. The preparation method thereof comprises the following steps:

[0170] (S1) APG, stearic acid, castor oil, monoglyceride, dimethyl silicone oil (100 cs), polydimethylsiloxane and coconut oil were stirred and dissolved uniformly at 75°C and 350 r / min, and the mixture was kept warm for 10 min. The mixture was recorded as phase A and set aside.

[0171] (S2) The temperature was raised to 90° C., 1,3-propylene glycol, glycerol, hyaluronic acid and deionized water were stirred and mixed at 300 r / min, maintained for 15 min, and then cooled to 75° C., which was recorded as phase B.

[0172] (S3) Slowly add phase B into phase A at 75°C and 300 r / min and stir until uniform.

[0173] (S4) When the temperature is cooled to 40° C. under stirring, xanthan gum and hydroxyethyl cellulose are added and stirred evenly. After they are completely dissolved, heating is stopped, and the mixture is cooled to room temperature under stirring. The preparation is completed, which is Example 13;

[0174] (S5) After the above (S4) was completely dissolved, 1% celery seed oil, ethyl acetate phase of jujube leaves and a composite of ethyl acetate phase containing jujube leaves (celery seed oil: ethyl acetate phase of jujube leaves = 1:11) were added respectively, stirred and cooled to room temperature, and they were defined as Example 14, Example 15 and Example 16 respectively. The changes in their properties were observed, and the results are shown in Table 8.

[0175] Table 8 Effects of different extract samples on creams

[0176]

[0177]

[0178] Note: The pH (25°C) of creams in the current industry standards meets the requirements within the range of 4.0 to 8.5.

[0179] From Table 8 and Figure 7 It can be seen that the three plant extracts do not have a significant effect on the appearance and physicochemical properties of the cream formula. The absorption effect of the cream containing celery seed oil (Examples 14 and 16) becomes better, and the shelf life of the formulas with added extract samples (celery seed oil, ethyl acetate phase of Ziziphus jujuba leaves and composites containing ethyl acetate phase of Ziziphus jujuba leaves) is extended. The cream with the composite containing ethyl acetate phase of Ziziphus jujuba leaves has the longest shelf life, which increases to 83 days. This may be related to the good antibacterial effect of the composite containing ethyl acetate phase of Ziziphus jujuba leaves.

[0180] In summary, the ethyl acetate phase compound containing jujube leaves, which has synergistic antioxidant, whitening, antibacterial and other functions, was added to the gel mask and cream with good texture and appearance. The physical and chemical properties and sensory evaluation were carried out according to the current industry standards, and all met the requirements, and its properties had no adverse effects on the blank group. After adding the ethyl acetate phase compound containing jujube leaves to the cream and mask, celery seed oil was conducive to the penetration of the functional substances, and the shelf life of the prepared product was significantly extended. The shelf life of the gel mask can be extended to 3 times that of the blank group; the shelf life of the cream is extended to 2.3 times that of the blank group.

[0181] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the explanation of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. A composite material containing ethyl acetate phase of wild jujube leaves, characterized in that: The following components are included in mass percentage: Celery seed oil: jujube leaf ethyl acetate phase = 1:0.1~15.

2. The composite material containing ethyl acetate phase of wild jujube leaves according to claim 1, characterized in that: The celery seed oil is prepared by the following method: The celery seeds are crushed and soaked in a sodium chloride aqueous solution, and then heated in an oil bath and post-treated to obtain celery seed oil; The material-liquid ratio of celery seeds to sodium chloride aqueous solution is 1:25-30; During the soaking process, the temperature is room temperature and the time is 3 to 4 hours; During the oil bath heating process, the temperature is 130-140°C and the time is 4-5 hours.

3. The composite material containing ethyl acetate phase of wild jujube leaves according to claim 1, characterized in that: The ethyl acetate phase of sour jujube leaves is prepared by the following method: The wild jujube leaves are crushed and placed in an ethanol solution, heated under reflux, and then post-processed to obtain a wild jujube leaf alcohol extract, and then the wild jujube leaf alcohol extract is dissolved in distilled water, and then extracted with ethyl acetate, and the upper layer solution is evaporated to remove the solvent to obtain a wild jujube leaf ethyl acetate phase; The solid-liquid ratio of the sour jujube leaves to the ethanol solution is 1:10-15; During the reflux process, the temperature is 85-95°C and the time is 2-3h; The usage ratio of distilled water to the alcohol extract of Ziziphus jujuba leaves is 40-60 mL:1 g, and the usage of ethyl acetate is the same as that of distilled water.

4. The composite material containing ethyl acetate phase of wild jujube leaves according to claim 1, characterized in that: The following components are included in mass percentage: Celery seed oil: jujube leaf ethyl acetate phase = 1:0.2~13.

5. The composite material containing ethyl acetate phase of wild jujube leaves according to claim 1, characterized in that: The following components are included in mass percentage: Celery seed oil: ethyl acetate phase of jujube leaves = 1:5; Celery seed oil: ethyl acetate phase of jujube leaves = 1:9; Celery seed oil: jujube leaf ethyl acetate phase = 1:

11.

6. The composite material containing ethyl acetate phase of wild jujube leaves according to claim 1, characterized in that: The following components are included in mass percentage: Celery seed oil: jujube leaf ethyl acetate phase = 1:

11.

7. A method for preparing a composite material containing ethyl acetate phase of wild jujube leaves as claimed in any one of claims 1 to 6, characterized in that: The following steps are involved: The celery seed oil and the ethyl acetate phase of the wild jujube leaves are mixed evenly to obtain a composite material containing the ethyl acetate phase of the wild jujube leaves.

8. Use of the ethyl acetate phase compound containing wild jujube leaves as claimed in any one of claims 1 to 6 in the preparation of facial masks and / or creams with synergistic antibacterial, anti-oxidation, whitening, and penetration-promoting functions.

9. A facial mask, characterized in that: The invention comprises the ethyl acetate phase compound containing jujube leaves as described in any one of claims 1 to 6.

10. A cream, characterized in that: The invention comprises the ethyl acetate phase compound containing jujube leaves as described in any one of claims 1 to 6.

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