Preparation method of waxberry extract as well as product and application of waxberry extract

Abstract

The invention relates to a preparation method of a waxberry extract as well as a product and application of the waxberry extract. The preparation method comprises the following steps: mixing a waxberry raw material with a compound enzyme for enzymolysis, adding a solution containing a deep-eutectic solvent into an enzymolysis reaction solution after enzymolysis is completed, performing high-voltage pulsed electric field extraction, centrifugation and filtration, and collecting filtrate to obtain the waxberry extract, the compound enzyme comprises a combination of cellulase, pectinase and ficin. According to the method, a high-voltage pulsed electric field is combined with a deep eutectic solvent to carry out synergistic extraction on a waxberry treating fluid subjected to enzymolysis, effective components in waxberries can be efficiently extracted, a specific enzyme combination is further adopted, and it is found that the extraction effect can be synergistically improved through combination of three kinds of enzymes; the prepared waxberry extract has excellent moisturizing and antioxidant effects, and is suitable for skin care products.

Description

Technical Field

[0001] The invention belongs to the technical field of plant extracts and relates to a preparation method of a bayberry extract and a product and application thereof. Background Art

[0002] Bayberry (Morella rubra Lour.) is an evergreen tree of the genus Myrica in the Myricaceae family. The crown is neat and spherical, and the branches are brittle and easy to break; the leaves are leathery, alternate, lanceolate or obovate; dioecious or occasionally monoecious; the female inflorescence is a catkin, which is bright red and filamentous; the fruit is a drupe-shaped ball with nipple-like protrusions on the outer surface; the fruit color is red, purple, white, pink, etc.; it likes warmth and is cold-resistant, cannot tolerate extreme heat, likes humidity and tolerates shade; Bayberry is also known as Longqing and Zhuhong, because it looks like hawthorn and tastes like plum, so it is named bayberry.

[0003] Bayberry has high medicinal value. The Compendium of Materia Medica records that "salt can be used to store food, eliminate phlegm and stop vomiting, and swallowing juice can benefit the five internal organs and lower the qi". It has the effects of harmonizing the stomach and digesting food, relieving heat and stopping diarrhea. Bayberry is rich in various nutrients, such as vitamin C, vitamin E and flavonoids. It also contains some antioxidants and anti-inflammatory substances such as myricetin, which have significant antioxidant, free radical elimination, soothing, prevention of vascular inflammation and cholesterol reduction effects. Myricetin is a natural skin care ingredient. It is rich in vitamin C and is a powerful antioxidant that can neutralize free radicals and prevent them from damaging skin cells. In addition, myricetin can also promote the synthesis of collagen in the skin, which is a key protein to maintain skin elasticity and firmness. Therefore, myricetin can help improve skin quality, nourish and protect the skin, and prevent and alleviate symptoms related to skin problems. Therefore, it is very necessary to provide a new, safe, environmentally friendly and efficient preparation method for bayberry extract. Summary of the invention

[0004] In view of the deficiencies of the prior art, the object of the present invention is to provide a method for preparing a bayberry extract and its product and application.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a method for preparing a bayberry extract, the preparation method comprising the following steps:

[0007] The bayberry raw material is mixed with a composite enzyme for enzymolysis, and after the enzymolysis is completed, a solution containing a low eutectic solvent is added to the enzymolysis reaction liquid, and high-voltage pulse electric field extraction is performed, and after centrifugation and filtration, the filtrate is collected to obtain the bayberry extract;

[0008] The complex enzyme comprises a combination of cellulase, pectinase and ficin.

[0009] The present invention creatively discovers that the composite enzyme composed of three enzymes, cellulase, pectinase and ficin, can interact with each other and positively influence each other, thereby improving the extraction efficiency of bayberry raw materials and thus increasing the active ingredients in the extract product.

[0010] In addition, the present invention adopts a high-voltage pulse electric field combined with a low eutectic solvent to synergistically extract the bayberry treated liquid after enzymatic hydrolysis, which can efficiently extract the effective ingredients in the bayberry, prepare a highly active bayberry extract product, and further apply it to cosmetics to improve the moisturizing and antioxidant effects of the product.

[0011] Preferably, the mass ratio of the cellulase, pectinase and ficin is (1-5):(2-6):(0.5-2);

[0012] The specific values ​​of "1-5" mentioned above may be, for example, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, etc.;

[0013] The specific values ​​in the above “2-6” may be, for example, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, etc.;

[0014] The specific numerical value in the above "0.5-2" can be, for example, 0.5, 0.8, 1, 1.2, 1.5, 1.8, 2, etc. Other specific point values ​​within the numerical range can be selected, and they will not be described one by one here.

[0015] In the present invention, when the dosage ratio of each enzyme in the composite enzyme combination is within the above-mentioned specific range, the enzymatic extraction effect on bayberry is better.

[0016] Preferably, the bayberry raw material is mixed with water before the enzymolysis, and the solid-to-liquid ratio is 1: (15-50) g / mL, for example, it can be 1: 15g / mL, 1: 18g / mL, 1: 20g / mL, 1: 22g / mL, 1: 25g / mL, 1: 28g / mL, 1: 30g / mL, 1: 32g / mL, 1: 35g / mL, 1: 38g / mL, 1: 40g / mL, 1: 42g / mL, 1: 45g / mL, 1: 50g / mL, etc. Other specific point values ​​within this numerical range can be selected, and they will not be repeated here one by one.

[0017] In the present invention, when the material-liquid ratio of the bayberry raw material and water is within the above-mentioned specific range, a better enzymatic hydrolysis effect is achieved.

[0018] Preferably, the added amount of the complex enzyme is 0.1‰-0.5‰ of the mass of the bayberry raw material, for example, it can be 0.1‰, 0.15‰, 0.2‰, 0.25‰, 0.3‰, 0.35‰, 0.4‰, 0.45‰, 0.5‰, etc. Other specific point values ​​within this numerical range can be selected, which will not be repeated here.

[0019] In the present invention, when the added amount of the complex enzyme is within the above specific range, the extraction effect on the bayberry raw material is better.

[0020] Preferably, the temperature of the enzymatic hydrolysis is 40-60°C, for example, it can be 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 52°C, 55°C, 58°C, 59°C, 60°C, etc.; the time is 1-3h, for example, it can be 1h, 1.2h, 1.5h, 1.8h, 2h, 2.3h, 2.5h, 2.8h, 3h, etc. Other specific point values ​​within this numerical range can be selected, which will not be repeated here.

[0021] Preferably, the deep eutectic solvent is obtained by reacting a hydrogen bond acceptor and a hydrogen bond donor.

[0022] Preferably, the hydrogen bond donor is selected from any one or a combination of at least two of propylene glycol, glycerol, glycerol, glucose, glycolic acid, citric acid, L-ascorbic acid, polyethylene glycol, fructose or L-proline, preferably a combination of propylene glycol and glycerol.

[0023] Preferably, the hydrogen bond acceptor is selected from any one or a combination of at least two of betaine, choline chloride or acetamide, preferably betaine.

[0024] Preferably, the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is (2-8):(1-3), for example, it can be 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 2:3, etc. Other specific point values ​​within this numerical range can be selected and will not be described one by one here.

[0025] Preferably, the solution containing the low eutectic solvent also includes water, and the mass ratio of the low eutectic solvent to water in the mixed solvent is 1:1-10:1, for example, it can be 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described one by one here.

[0026] Preferably, the ratio of the bayberry raw material to the solution containing the low eutectic solvent is 1: (4-12) g / mL, for example, it can be 1: 4g / mL, 1: 5g / mL, 1: 6g / mL, 1: 7g / mL, 1: 8g / mL, 1: 9g / mL, 1: 10g / mL, 1: 11g / mL, 1: 12g / mL, etc., and other specific point values ​​within this numerical range can be selected, which will not be repeated here.

[0027] Preferably, the field strength of the high-voltage pulse electric field is 250-420kv / m, for example, it can be 250kv / m, 280kv / m, 300kv / m, 320kv / m, 350kv / m, 380kv / m, 400kv / m, 420kv / m, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described here one by one.

[0028] Preferably, the number of electric shocks of the high-voltage pulse electric field is 50-180 times, for example, it can be 50 times, 80 times, 100 times, 120 times, 150 times, 160 times, 180 times, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described one by one here.

[0029] In the present invention, the high-voltage pulse electric field extraction using the field strength and electric shock times within the above-mentioned specific parameter range can not only gently inactivate the enzymes remaining in the enzymatic hydrolysis process, but also further extract the active ingredients in the bayberry, thereby achieving a better extraction effect.

[0030] In a second aspect, the present invention provides a bayberry extract, which is prepared by the preparation method described in the first aspect.

[0031] In a third aspect, the present invention provides a use of the bayberry extract according to the second aspect in cosmetics.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] The present invention develops a new preparation method of bayberry extract, creatively adopts enzymatic hydrolysis and high-voltage pulse electric field technology combined with a low eutectic solvent to extract the bayberry raw material, and further adopts a composite enzyme combination including cellulase, pectinase and ficin for enzymatic hydrolysis. The three enzymes are used in combination, which can interact and positively affect each other, thereby improving the extraction efficiency of bayberry, thereby improving the active ingredients in the extract product. The extraction method of the present invention can efficiently extract the effective ingredients in bayberry, prepare a highly active bayberry extract product, and can be further applied to cosmetics to improve the moisturizing and anti-oxidation effects of the product. DETAILED DESCRIPTION

[0034] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0035] The cellulase, pectinase and ficin involved in the following examples are purchased from Shanghai Yuanye Biotechnology Co., Ltd., and the models are S10041, S10007 and V30095 respectively.

[0036] Example 1

[0037] This embodiment provides a bayberry extract, and the preparation method thereof is as follows:

[0038] (1) The washed and dried bayberry slurry was crushed until there were no obvious large particles, and the bayberry crushed material was mixed with pure water at a solid-liquid ratio of 1:30 g / mL, and a composite enzyme (composed of cellulase, pectinase and ficin in a mass ratio of 3:4:1) was added for enzymatic hydrolysis at a temperature of 50°C for 2 h. The amount of the composite enzyme added was 0.3‰ of the bayberry crushed material, and an enzymatic hydrolyzate was obtained after the reaction was completed.

[0039] (2) adding a solution containing a low eutectic solvent [(propylene glycol + glycerol): betaine = 3:1, propylene glycol: glycerol = 1:6, molar ratio] to the enzymatic hydrolyzate and mixing for leaching, wherein the mass ratio of the low eutectic solvent to water in the solution is 5:1, and the solid-liquid ratio of the bayberry crushed material to the solution containing the low eutectic solvent is 1:8 g / mL, and the leaching is carried out under the condition of a high-voltage pulse electric field, the leaching condition is 350 kv / m, the number of electric shocks is 150 times, and after the extract is centrifuged, the supernatant is filtered (three-layer gauze), and the filtrate is collected and freeze-dried to obtain the bayberry extract.

[0040] Example 2

[0041] This embodiment provides a bayberry extract, and the preparation method thereof is as follows:

[0042] (1) The washed and dried bayberry slurry was crushed until there were no obvious large particles, and the bayberry crushed material was mixed with pure water at a solid-liquid ratio of 1:20 g / mL, and a composite enzyme (composed of cellulase, pectinase and ficin in a mass ratio of 1:5:0.5) was added for enzymatic hydrolysis at a temperature of 45°C for 2.6 h, wherein the amount of the composite enzyme added was 0.1‰ of the bayberry crushed material, and an enzymatic hydrolyzate was obtained after the reaction was completed.

[0043] (2) adding a solution containing a low eutectic solvent [(fructose + L-proline): choline chloride = 1:1, fructose: L-proline = 1:1, molar ratio] to the enzymatic hydrolyzate and mixing for leaching, wherein the mass ratio of the low eutectic solvent to water in the solution is 8:1, and the solid-liquid ratio of the bayberry crushed material to the solution containing the low eutectic solvent is 1:5 g / mL, and the leaching is carried out under the condition of a high-voltage pulse electric field, the leaching condition is 250 kv / m, the number of electric shocks is 180 times, and after the extract is centrifuged, the supernatant is filtered (three-layer gauze), and the filtrate is collected and freeze-dried to obtain the bayberry extract.

[0044] Example 3

[0045] This embodiment provides a bayberry extract, and the preparation method thereof is as follows:

[0046] (1) The washed and dried bayberry slurry was crushed until there were no obvious large particles, and the bayberry crushed material was mixed with pure water at a solid-liquid ratio of 1:45 g / mL, and the mixture was hydrolyzed at 58°C for 1.2 h with a composite enzyme (composed of cellulase, pectinase and ficin in a mass ratio of 5:2:0.8). The amount of the composite enzyme added was 0.4‰ of the bayberry crushed material, and an enzymatic solution was obtained after the reaction was completed.

[0047] (2) adding a solution containing a low eutectic solvent [(glucose + glycolic acid): betaine = 2:3, glucose: glycolic acid = 3:2, molar ratio] to the enzymatic hydrolyzate and mixing for leaching, wherein the mass ratio of the low eutectic solvent to water in the solution is 2:1, and the solid-liquid ratio of the bayberry crushed material to the solution containing the low eutectic solvent is 1:12 g / mL, and the leaching is carried out under the condition of a high-voltage pulse electric field, the leaching condition is 420 kv / m, the number of electric shocks is 50 times, and after the extract is centrifuged, the supernatant is filtered (three-layer gauze), and the filtrate is collected and freeze-dried to obtain the bayberry extract.

[0048] Example 4

[0049] This embodiment provides a bayberry extract, which uses the crushed bayberry of Example 1. The only difference between its preparation method and that of Example 1 is that the amount of compound enzyme added is 2‰, and the remaining steps and process parameters are all referenced to Example 1.

[0050] Example 5

[0051] This embodiment provides a bayberry extract, which uses the bayberry crushed material of Example 1. The only difference between its preparation method and that of Example 1 is that the addition amount of the complex enzyme is 0.05‰, and the remaining steps and process parameters are all referred to Example 1.

[0052] Example 6

[0053] This embodiment provides a bayberry extract, which uses the enzymatic hydrolyzate after the reaction in Example 1. The only difference between its preparation method and that in Example 1 is that the field strength of the high-voltage electric field is different, which is 200 kv / m. The remaining steps and process parameters are all referred to in Example 1.

[0054] Example 7

[0055] This embodiment provides a bayberry extract, which uses the enzymatic hydrolyzate after the reaction in Example 1. The only difference between its preparation method and that in Example 1 is that the field strength of the high-voltage electric field is different, which is 450 kv / m. The remaining steps and process parameters are all referenced to Example 1.

[0056] Example 8

[0057] This embodiment provides a bayberry extract, which uses the enzymatic hydrolyzate after the reaction in Example 1. The preparation method thereof is different from that in Example 1 only in that the number of electric shocks in the high-voltage electric field is different, which is 30 times. The remaining steps and process parameters are all referred to Example 1.

[0058] Example 9

[0059] This embodiment provides a bayberry extract, which uses the enzymatic hydrolyzate after the reaction in Example 1. The preparation method thereof is different from that in Example 1 only in that the number of electric shocks in the high-voltage electric field is different, which is 200 times. The remaining steps and process parameters are all referred to Example 1.

[0060] Comparative Example 1

[0061] This comparative example provides a bayberry extract, which uses the bayberry crushed material of Example 1. The only difference between its preparation method and that of Example 1 is that the complex enzyme does not contain cellulase, and its reduced amount is proportionally distributed to pectinase and ficin. The remaining steps and process parameters are all referred to Example 1.

[0062] Comparative Example 2

[0063] This comparative example provides a bayberry extract, which uses the bayberry crushed material of Example 1. The only difference between its preparation method and that of Example 1 is that the complex enzyme does not contain pectinase, and its reduced portion is proportionally distributed to cellulase and ficin. The remaining steps and process parameters are all referred to Example 1.

[0064] Comparative Example 3

[0065] This comparative example provides a bayberry extract, which uses the bayberry crushed material of Example 1. The only difference between its preparation method and that of Example 1 is that the complex enzyme does not contain ficin, and its reduced portion is proportionally distributed to cellulase and pectinase. The remaining steps and process parameters are all referred to Example 1.

[0066] Comparative Example 4

[0067] This comparative example provides a bayberry extract, which uses the bayberry crushed material of Example 1. The preparation method thereof is different from that of Example 1 only in that no enzymatic hydrolysis is performed. After the bayberry crushed material is mixed with pure water, it is heated in a water bath at 50° C. for 2 h, and then step (2) is directly performed. The relevant treatment and process parameters are all referred to Example 1 to obtain the bayberry extract.

[0068] Comparative Example 5

[0069] This comparative example provides a bayberry extract, which uses the enzymatic hydrolysate after the reaction in Example 1. The preparation method thereof is different from that in Example 1 only in that high-voltage pulse electric field extraction is not performed, and the enzymatic hydrolysate is mixed with a solution containing a low eutectic solvent and then extracted. The extraction temperature is 80° C., the extraction time is 60 min, and the bayberry extract is obtained by centrifugation, filtering the supernatant, collecting the filtrate, and freeze-drying.

[0070] Comparative Example 6

[0071] This comparative example provides a bayberry extract, which uses the enzymatic hydrolyzate after the reaction in Example 1. The preparation method thereof is different from that in Example 1 only in that the solution containing the low eutectic solvent in step (2) is replaced with an equal volume of pure water, and high-voltage pulse electric field leaching and subsequent treatment are performed. The relevant process parameters refer to Example 1 to prepare the bayberry extract.

[0072] Test Example 1

[0073] Antioxidant effect test

[0074] (1) DPPH-free radical scavenging

[0075] A 96-well plate was used, and three replicates were set for each group, with a system of 200 μL. Sample group: 100 μg of the extract samples prepared in Examples 1-9 and Comparative Examples 1-6 were taken, respectively, mixed with 100 μL of pure water, and 100 μL of DPPH solution (0.1 mM) was added to the reaction system; control group: 100 μL of pure water was taken, and then 100 μL of DPPH solution (0.1 mM) was added. After the reaction system was constructed, it was shaken in the dark for 10 minutes, and the absorbance at 517 nm was tested using an ELISA instrument.

[0076] The calculation method of clearance rate is as follows: Clearance rate (%) = [(A0-Ax) / A0] × 100%, A0 is the absorbance of the control group, and Ax is the absorbance of the sample group.

[0077] (2) ABTS-free radical scavenging

[0078] Preparation of ABTS working solution: Mix ABTS and potassium persulfate solution to prepare 7 mM ABTS stock solution, store in dark for 12 h, shake well and mix with 10 mM PBS (pH=7.4) so ​​that the absorbance of the solution at 734 nm is 0.8, to obtain ABTS working solution.

[0079] A 96-well plate was used, with three replicates in each group, and the system was 200 μL. Sample group: 100 μg of the extract samples prepared in Examples 1-9 and Comparative Examples 1-6 were taken, mixed with 10 μL of pure water, and 190 μL of ABTS working solution was added to the reaction system; control group: 10 μL of pure water was taken, and then 190 μL of ABTS working solution was added. After the reaction system was constructed, it was fully shaken and incubated for 6 minutes, and the absorbance at 734 nm was tested with an enzyme marker.

[0080] The calculation method of clearance rate is as follows: Clearance rate (%) = [(A0-Ax) / A0] × 100%, A0 is the absorbance of the control group, and Ax is the absorbance of the sample group.

[0081] The test results are shown in Table 1.

[0082] Table 1

[0083] Group DPPH-free radical scavenging rate ABTS-free radical scavenging rate Example 1 98.16％ 96.32％ Example 2 95.32％ 94.06％ Example 3 94.19％ 93.79％ Example 4 92.76％ 91.24％ Example 5 91.28％ 89.96％ Example 6 91.75％ 90.06％ Example 7 91.44％ 89.94％ Example 8 93.01％ 92.03％ Example 9 92.32％ 91.46％ Comparative Example 1 88.76％ 86.54％ Comparative Example 2 87.49％ 85.79％ Comparative Example 3 86.16％ 84.23％ Comparative Example 4 87.53％ 86.32％ Comparative Example 5 86.92％ 85.14％ Comparative Example 6 86.65％ 85.26％

[0084] From the data in the table, we can see that

[0085] (1) It can be seen from Examples 1 to 3 that the bayberry extract prepared by the present invention has excellent DPPH-free radical and ABTS-free radical scavenging effects and exhibits significant antioxidant effects, among which Example 1 has the best effect.

[0086] (2) By comparing Example 1 with Examples 4-5, it can be seen that when the amount of the complex enzyme added is within a specific range, the antioxidant effect of the bayberry extract prepared therefrom is better.

[0087] (3) By comparing Example 1 with Examples 6-9, it can be seen that when the parameters of the electric field strength and the number of electric shocks of the high-voltage pulse electric field used in the extraction are selected within a specific range, the extraction effect on bayberry is better, and a bayberry extract with higher antioxidant effect is prepared.

[0088] (4) By comparing Example 1 with Comparative Examples 1-3, it can be seen that the present invention has an excellent enzymatic hydrolysis and extraction effect on bayberry by using the three enzymes of cellulase, pectinase and ficin in combination. When Comparative Examples 1-3 do not contain any of the three enzymes of cellulase, pectinase and ficin, the antioxidant effect of the bayberry extracts prepared therefrom is lower than that of Example 1, indicating that the three enzymes of cellulase, pectinase and ficin have a synergistic effect when applied to the extraction of active ingredients in bayberry.

[0089] (6) By comparing Example 1 with Comparative Examples 4-6, it can be seen that the present invention adopts enzymatic hydrolysis and low eutectic solvent combined with high-voltage pulsed electric field technology to extract the bayberry raw material, and the synergistic enhancement further improves the extraction effect of bayberry, increases the content of active ingredients in the final bayberry extract, and improves the antioxidant activity of the product.

[0090] Test Example 2

[0091] Moisturizing effect test

[0092] (1) In vitro testing

[0093] Take 0.5 mL of the diluted samples of Examples 1-9 and Comparative Examples 1-6 (the bayberry extract is mixed with pure water, and the final concentration of the extract in the diluted sample is 5%), apply them on a glass plate with 3M tape, record the mass m of the glass plate with 3M tape and the mass m0 after applying the sample, put them in a desiccator filled with a saturated potassium carbonate solution (relative humidity is 48%), weigh and record the data m after leaving for 2h and 8h t Calculate the moisturizing rate at this time, repeat three times and calculate the average value. The moisturizing rate calculation method is: Moisturizing rate (%) = (m t -m) / (m0-m)×100%, the results are shown in Table 2.

[0094] (2) Human trials

[0095] 120 subjects were selected, with an age range of 28-40 years old and an average age of 34 years old. They were randomly divided into 15 groups. The subjects uniformly cleaned the curved side of the left and right forearms. After gently blotting with a paper towel, each group of samples marked 6 areas of 3cm×3cm as the test area, with at least 2cm between each area, and the test results were statistically averaged. The test area was smeared with an appropriate amount of the diluted samples of Example 1-9 and Comparative Example 1-6 (the bayberry extract was mixed with pure water, and the final concentration of the extract in the diluted sample was 1%). Before using the sample and after using it for 2 hours, the subject data was arranged for collection, and skin parameters were collected each time using a skin test instrument (Corneometer CM825), and the skin stratum corneum moisture content growth rate (%) was statistically calculated, and the growth rate = (moisture content after using it for 2 hours-moisture content before use) / moisture content before use, and the results are shown in Table 2.

[0096] Table 2

[0097]

[0098]

[0099] It can be seen from the data in the table that the bayberry extract prepared by the preparation method provided by the present invention has excellent moisturizing effect and can maintain the moisture content of the stratum corneum of the skin. The moisturizing effect of Example 1 is better than that of Examples 4-9 and Comparative Examples 1-6, indicating that the preparation conditions of the specific bayberry extract adopted by the present invention have different degrees of influence on the moisturizing effect of the final extraction product.

[0100] Test Example 3

[0101] Product safety testing

[0102] According to the subject inclusion criteria, 45 females and 45 males were selected to participate in the test, and randomly divided into 9 groups (5 males and 5 females in each group) to receive the trial. Take an appropriate amount of the extract sample prepared in Examples 1-9 (the extract is mixed with pure water to adjust the extract concentration to 5%, and then tested) and place it in the drug chamber of the spot test tape. The spot tester with the sample is applied to the skin of the curved side of the forearm of the subject with medical tape for 24 hours. 30 minutes after removing the test spot tester, observe the skin reaction after the indentation disappears. For those with negative results, observe again 24 hours and 48 hours after the test, and record the skin reaction results. The reaction degree grading standard is as shown in the following table.

[0103]

[0104] The results show that the products prepared in Examples 1-9 showed negative reactions 24h and 48h after the test, and there was no irritation or erythema. It can be seen that the bayberry extract product prepared by the present invention has no irritation to the skin and no positive reaction, which proves that the product is highly safe, less irritating, and milder.

[0105] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0106] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0107] 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 further describe various possible combinations.

Claims

1. A method for preparing a bayberry extract, characterized in that: The preparation method comprises the following steps: The bayberry raw material is mixed with a composite enzyme for enzymolysis, and after the enzymolysis is completed, a solution containing a low eutectic solvent is added to the enzymolysis reaction solution, and high-voltage pulse electric field extraction is performed, and after centrifugation and filtration, the filtrate is collected to obtain the bayberry extract; The complex enzyme comprises a combination of cellulase, pectinase and ficin.

2. The preparation method according to claim 1, characterized in that: The mass ratio of the cellulase, pectinase and ficin is (1-5):(2-6):(0.5-2).

3. The preparation method according to claim 1, characterized in that: Before the enzymatic hydrolysis, the bayberry raw material is mixed with water, and the solid-liquid ratio is 1:(15-50) g / mL.

4. The preparation method according to claim 1, characterized in that: The added amount of the complex enzyme is 0.1‰-0.5‰ of the weight of the bayberry raw material; Preferably, the enzymatic hydrolysis temperature is 40-60°C and the time is 1-3h.

5. The preparation method according to claim 1, characterized in that: The deep eutectic solvent is obtained by reacting a hydrogen bond acceptor and a hydrogen bond donor; Preferably, the hydrogen bond donor is selected from any one or a combination of at least two of propylene glycol, glycerol, glycerol, glucose, glycolic acid, citric acid, L-ascorbic acid, polyethylene glycol, fructose or L-proline; Preferably, the hydrogen bond acceptor is selected from any one of betaine, choline chloride or acetamide, or a combination of at least two thereof.

6. The preparation method according to claim 5, characterized in that: The molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is (2-8):(1-3); Preferably, the solution containing the low eutectic solvent also includes water, and the mass ratio of the low eutectic solvent to water in the mixed solvent is 1:1-10:

1.

7. The preparation method according to claim 1, characterized in that: The ratio of the bayberry raw material to the solution containing the low eutectic solvent is 1:(4-12) g / mL.

8. The preparation method according to claim 1, characterized in that: The field strength of the high voltage pulse electric field is 250-420 kv / m; Preferably, the number of electric shocks of the high-voltage pulse electric field is 50-180 times.

9. A bayberry extract, characterized in that The bayberry extract is prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the bayberry extract according to claim 9 in cosmetics.

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