Preparation method and application of W1 / O / W2 double emulsion based on embedding brain polypeptide freeze-dried powder

Abstract

The application discloses a preparation method and application of a W1 / O / W2 type double emulsion based on embedding brain polypeptide freeze-dried powder. The preparation method comprises the following steps: taking soybean oil loaded with polyglycerol ricinoleate (PGPR) as an oil phase, taking an aqueous solution loaded with brain polypeptide freeze-dried powder as an inner water phase, mixing the oil phase and the inner water phase and carrying out homogenization treatment to obtain W1 / O primary emulsion; taking a solution loaded with Tween 80, xanthan gum and sucrose fatty acid ester as an outer water phase, mixing the W1 / O primary emulsion and the outer water phase and carrying out homogenization treatment, adding fruit juice concentrate, and after stirring, tanking and sterilization, fruit-flavored brain polypeptide oral liquid is obtained. The double emulsion can significantly improve the release and absorption of brain polypeptide in the intestinal tract, improve the oral bioavailability, and can be used for small intestine targeted delivery of hydrophilic bioactive substances. The obtained emulsion has good stability at room temperature. The obtained fruit-flavored brain polypeptide oral liquid has strong palatability, and greatly improves the consumer acceptance.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of food, and relates to a preparation method of a W1 / O / W2 double emulsion suitable for embedding brain polypeptide freeze-dried powder and application thereof. BACKGROUND

[0002] Brain polypeptide is mainly a mixture of polypeptide obtained by enzymolysis and quantitative ultrafiltration of pig brain and 16 animal-derived amino acids. It was first developed by Ebewe Company, and the active ingredient is N-PEP-12 (Windisch M, Hutter-Paier B, Grygar E, et al. Journal of Neural Transmission, 2005, 112(10): 1331-1343.), which is composed of 70% peptides and 30% free amino acids. Brain polypeptide has the functions of regulating and improving neuron metabolism, activating brain neuron regeneration, promoting synapse formation, preventing neuron apoptosis, improving the functions of brain and cognitive, language and motor related regions, and protecting nerve cells from damage by various ischemia and neurotoxins. During the process of enzymatic hydrolysis of protein to form polypeptide, the hydrophobic amino acids are often exposed to directly bind with bitter receptors in the oral cavity, resulting in bitterness. Due to its poor sensory properties and easy degradation in the gastrointestinal tract, its wide application as a functional ingredient is limited. At the same time, the unsatisfactory sensory experience reduces the consumer's acceptance of peptide food.

[0003] Chinese patent CN103445263A discloses an enzymolysis egg white protein complex nutritional oral liquid. Egg white is subjected to enzymolysis to obtain small molecule polypeptides with biological activities such as antioxidant, anti-fatigue and blood pressure lowering, and then the enzymolysis egg white protein complex oral nutritional liquid with rich nutrition, enhanced immunity and suitable taste for adults is prepared by adding Chinese wolfberry extract. In the prior art, the enzymolysis egg white protein is mainly compounded with plant extracts to achieve the effects of antioxidant and immune enhancement, but there are still problems of poor product stability and palatability in the processing process.

[0004] In order to maintain the biological activity of brain polypeptide and the stability of small molecule polypeptide during storage, processing and intake, and finally obtain a product meeting the taste of consumers, the present application adopts the method of constructing a water-in-oil-in-water (W1 / O / W2) double emulsion to mask the bitterness of brain polypeptide, improve its bad taste and improve the consumer's acceptance.

[0005] Chinese patent CN 117838633 B discloses a method for preparing and applying a W1 / O / W2 type dual emulsion based on IgY encapsulation. The method uses a mixed solution of butter and corn oil loaded with polyglycerol ricinoleate (PGPR) as the oil phase and an IgY-loaded solution as the inner aqueous phase. The oil and aqueous phases are then mixed and sheared to obtain a W1 / O emulsion. A polysaccharide dispersion, serving as the outer aqueous phase, is mixed with the W1 / O emulsion and sheared to obtain a dual emulsion with co-encapsulated IgY, which improves the intestinal adhesion and survival rate of IgY in the gastrointestinal tract. However, the IgY release rate of the dual emulsion prepared by this method is only 40% when exposed to simulated intestinal fluid, and the release rate needs further improvement to facilitate nutrient absorption.

[0006] Dual emulsion carriers offer advantages such as flavor masking, controlled release of active ingredients, and improved stability. However, in actual production, the emulsifiers used in dual emulsions can introduce unpleasant flavors, affecting the product's taste. Oral liquid products not only require high nutritional value but also good sensory quality and palatability. Therefore, this invention further optimizes the flavor of W1 / O / W2 type dual emulsions encapsulating lyophilized brain peptide powder to address issues of poor palatability and monotonous flavor. Furthermore, adding concentrated fruit juice to the W1 / O / W2 type dual emulsion significantly masks unpleasant flavors, enhances the beverage's sensory quality, and the resulting emulsion does not separate after three months of storage at room temperature. Summary of the Invention

[0007] In view of the problems existing in the prior art, the present invention provides a method for preparing a W1 / O / W2 type dual emulsion using lyophilized brain polypeptide powder as raw material and its application, aiming to solve some of the problems in the prior art or at least alleviate some of the problems in the prior art.

[0008] This invention provides a method for preparing lyophilized brain polypeptide powder.

[0009] The brain protein hydrolysate was placed in a freeze dryer and freeze-dried under vacuum to obtain brain polypeptide lyophilized powder.

[0010] Further specifying the preparation method of the brain polypeptide lyophilized powder, the method is as follows: 0.5g of porcine brain protein powder is dissolved in 15mL of ultrapure water, 0.1g of trypsin is added for enzymatic hydrolysis for 2h, the enzyme is inactivated by boiling water bath for 10min, the pH is adjusted to 7.0, and the mixture is filtered to obtain the brain protein hydrolysate.

[0011] Further specifying, the method for obtaining the pig brain protein powder is as follows:

[0012] Thaw frozen pig brains (with fascia removed) and crush them using a high-speed blender to obtain a pig brain homogenate. Mix the pig brain homogenate with edible alcohol at a mass ratio of 1:2, and extract in a 78℃ water bath with stirring for 2 hours. Let it stand for 10 minutes, and filter the extract to obtain a filter cake. Repeat the above operation 5 times for a total of 10 hours. Place the final filter cake in a 45℃ vacuum drying oven and dry for 12 hours to obtain pig brain protein powder.

[0013] This invention develops a novel emulsion system for encapsulating lyophilized brain polypeptide powder, which solves the problem of easy inactivation of lyophilized brain polypeptide powder in the gastrointestinal tract, improves the release rate and storage stability of lyophilized brain polypeptide powder in the intestine, and improves the bitterness and fishy taste of the enzymatic hydrolysate.

[0014] The first aspect of this invention is to provide a method for preparing a highly stable W1 / O / W2 type dual emulsion for encapsulating lyophilized brain peptide powder, comprising the following steps:

[0015] S1: Preparation of inner aqueous phase W1: After mixing the lyophilized brain peptide powder with ultrapure water, the mixture is vortexed to disperse it evenly, and then filtered through a 0.22μm filter membrane to form inner aqueous phase W1;

[0016] S2: Preparation of oil phase O: Dissolve a lipophilic emulsifier in edible oil and stir magnetically to form oil phase O;

[0017] S3: Preparation of colostrum W1 / O: The oil phase O and the inner aqueous phase W1 are initially mixed, and then homogenized by stirring, homogenizing and ultrasonication to obtain water-in-oil colostrum W1 / O.

[0018] S4: Preparation of external aqueous phase W2: Add hydrophilic emulsifier and sucrose fatty acid ester to ultrapure water, stir and dissolve in a 50℃ water bath for 30 min to obtain external aqueous phase W2;

[0019] S5: Preparation of W1 / O / W2 type dual emulsion: The primary emulsion W1 / O is added dropwise to the external aqueous phase W2, and after stirring, high-speed shearing, homogenization emulsification and ultrasonic emulsification, the W1 / O / W2 type dual emulsion encapsulating brain polypeptide lyophilized powder can be obtained.

[0020] S6: Preparation of highly stable W1 / O / W2 type dual emulsion: Add stabilizer to the W1 / O / W2 type dual emulsion, and after stirring, homogenization and ultrasonic emulsification, a highly stable W1 / O / W2 type dual emulsion can be obtained.

[0021] In one specific embodiment, the loading of the brain polypeptide lyophilized powder in the inner aqueous phase W1 in step S1 is 25 mg / mL.

[0022] In another specific embodiment, the edible oil mentioned in step S2 is soybean oil.

[0023] In another specific embodiment, the lipophilic emulsifier in step S2 includes, but is not limited to, polyglycerol ricinoleate (PGPR), Span 80, mono- and diglycerides of fatty acids, soybean lecithin, modified soybean lecithin, lactic acid fatty acid glycerides, etc.; preferably, polyglycerol ricinoleate (PGPR).

[0024] In another specific embodiment, the amount of lipophilic emulsifier added in step S2 is 1% to 10% of the mass of oil phase O; most preferably 6%.

[0025] In another specific embodiment, step S3 involves adding the inner aqueous phase W1 to the oil phase O while stirring, wherein the volume ratio of the oil phase O to the inner aqueous phase W1 is one of 5:5, 6:4, 7:3, 8:2, or 9:1.

[0026] In another specific embodiment, in S4, W2 is prepared by adding a hydrophilic emulsifier and sucrose fatty acid ester to deionized water and stirring until fully dissolved. The hydrophilic emulsifier includes, but is not limited to, at least one of sodium carboxymethyl cellulose (CMC-Na), gum arabic, modified soybean lecithin, soluble soybean polysaccharide, Tween 80, and sucrose fatty acid ester; most preferably, the hydrophilic emulsifier is Tween 80.

[0027] In another specific embodiment, the concentration of the hydrophilic emulsifier in W2 in step S4 is 1-10%, preferably 4%; the concentration of the sucrose fatty acid ester is 0.1%; and the dissolution is carried out by stirring at 50°C for 30 minutes.

[0028] In another specific embodiment, in step S5, the volume ratio of the external aqueous phase W2 to the internal phase W1 / O is 5:5, 6:4, 7:3, 8:2, or 9:1; preferably 6:4.

[0029] In another specific embodiment, the stabilizer mentioned in step S6 is xanthan gum.

[0030] In another specific embodiment, the concentration of the stabilizer in W2 mentioned in step S6 is 0.2-1%; preferably 0.6-1%, and more preferably 0.8%.

[0031] A second aspect of the present invention also provides a highly stable W1 / O / W2 type dual emulsion based on lyophilized brain polypeptide powder, characterized in that it is prepared by any of the methods described in the first aspect of the present invention.

[0032] The third aspect of this invention is to provide the application of the highly stable W1 / O / W2 type dual emulsion based on encapsulated brain polypeptide lyophilized powder described in the second aspect in the preparation of fruit-flavored brain polypeptide oral liquid; the preparation steps are as follows:

[0033] a. Adjust the pH of the above-mentioned highly stable W1 / O / W2 type dual emulsion to 7.0;

[0034] b. Mix the highly stable W1 / O / W2 type double emulsion and fruit juice concentrate in a certain proportion, stir evenly, and adjust the pH to 7.0;

[0035] c. Filter the emulsion obtained in step b through a 500-mesh filter cloth, and then fill it into containers;

[0036] d. The emulsion obtained in step c is subjected to high temperature and high pressure sterilization. The sterilization method is: high pressure steam sterilization at 121℃ for 15 minutes, and cooling to the core temperature of less than 35℃ to obtain fruit flavor brain polypeptide oral liquid.

[0037] e. Sensory evaluation of the above-mentioned fruit-flavored brain polypeptide oral liquid.

[0038] In one specific embodiment, the fruit juice concentrate is one or more of apple concentrate, jujube concentrate, peach concentrate, mango concentrate, grape concentrate, lemon concentrate, and orange concentrate.

[0039] The advantages of this invention are:

[0040] (1) The established W1 / O / W2 system improves the sensitivity of brain peptides to gastrointestinal digestion, and functional components can be encapsulated in the internal aqueous phase and released into the intestine at a controlled rate.

[0041] (2) A W1 / O / W2 system with high encapsulation efficiency and high stability, suitable for encapsulating lyophilized brain peptide powder, was successfully established. The formulation and preparation method of this invention, by compounding Tween 80 and xanthan gum in a specific ratio, helps to improve the stability of the W1 / O / W2 emulsion, which does not separate after being placed at room temperature for 3 months.

[0042] (3) This study utilizes highly stable W1 / O / W2 type double emulsion to develop fruit-flavored peptide beverages, masking their unpleasant flavors. The products have superior sensory quality, strong palatability, and improved consumer acceptance.

[0043] (4) The fruit-flavored brain polypeptide oral liquid provided by the present invention is easy to take, has a good color and is rich in nutrients, which expands the application of brain polypeptide in the field of health drinks and increases the added value of deep processing of pig brain.

[0044] (5) The preparation conditions of this invention are mild, the preparation process is simple, the raw materials are readily available, and it is suitable for large-scale production.

[0045] (6) The oral liquid of the present invention is more convenient to carry and easier to absorb after being filled. High-pressure sterilization can extend the shelf life and make the oral liquid more hygienic and safe. Attached Figure Description

[0046] Figure 1 This is an HPLC chromatogram of brain protein hydrolysate.

[0047] Figure 2 The image shows the appearance of the W1 / O / W2 type dual emulsion prepared in Example 8. The xanthan gum content from left to right is 0%, 0.2%, 0.4%, 0.6%, 0.8%, and 1.0%.

[0048] Figure 3 The image shows the microscopic morphology of the W1 / O / W2 type double emulsion prepared in Example 8 under an optical microscope. The xanthan gum addition amounts from a to f were 0%, 0.2%, 0.4%, 0.6%, 0.8%, and 1.0%, respectively.

[0049] Figure 4 The embedding rate of the W1 / O / W2 type dual emulsion prepared in Example 8.

[0050] Figure 5 Emulsifying activity (EAI) and emulsifying stability (ESI) of the W1 / O / W2 type dual emulsion prepared in Example 8.

[0051] Figure 6 The storage TSI value of the W1 / O / W2 type dual emulsion prepared in Example 8.

[0052] Figure 7 Centrifugal retention rate of the W1 / O / W2 type dual emulsion prepared in Example 8.

[0053] Figure 8 The free fatty acid (FFA) release rate of the W1 / O / W2 type dual emulsion prepared in Example 8 after in vitro simulation of gastrointestinal fluid.

[0054] Figure 9 The chemical stability (S) of the W1 / O / W2 type dual emulsion prepared in Example 8 after in vitro simulation of gastrointestinal fluid was determined. * ) and biological accessibility (B * ). Detailed Implementation

[0055] The objects and functions of the present invention, as well as the methods for achieving these objects and functions, will be explained below with reference to exemplary embodiments. However, the present invention is not limited to the exemplary embodiments disclosed below; it can be implemented in various forms. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0056] Example 1: Preparation and Detection of Lyophilized Brain Polypeptide Powder

[0057] 1. Preparation of porcine brain protein powder

[0058] Thaw frozen pig brains (with fascia removed) and crush them using a high-speed blender to obtain a homogenate. Mix the pig brain homogenate with edible alcohol at a ratio of 1:2, and extract by stirring in a 78℃ water bath for 2 hours. Let it stand for 10 minutes, then filter the extract to obtain a filter cake. Repeat the above operation 5 times for a total of 10 hours. Place the final filter cake in a 45℃ vacuum drying oven and dry for 12 hours to obtain pig brain protein powder.

[0059] 2. Detection of nitrogen content in porcine brain protein powder

[0060] Take 0.200g of dried pig brain protein powder, add two digestive tablets (4g / tablet), slowly add 12mL of concentrated sulfuric acid, digest for 3h, and determine the nitrogen content using a FOSS 8400 fully automatic Kjeldahl nitrogen analyzer. The results are shown in Table 1.

[0061] 3. Preparation of brain protein hydrolysate

[0062] Dissolve 1g of porcine brain protein powder in 15mL of ultrapure water, add 0.3g of trypsin and hydrolyze at 50℃ for 1h. Inactivate the enzyme in a boiling water bath for 10min, adjust the pH to 7.0, filter to remove residue, and collect the filtrate to obtain brain protein hydrolysate.

[0063] 4. Detection of nitrogen content and molecular weight in brain protein hydrolysate

[0064] (1) Take 2 mL of brain protein hydrolysate, add two digestion tablets (4 g / tablet), slowly add 12 mL of concentrated sulfuric acid, digest for 3 h, and determine the nitrogen content using a FOSS8400 fully automatic Kjeldahl nitrogen analyzer. The results are shown in Table 1.

[0065] (2) The molecular weight of the brain protein hydrolysate was determined by high performance liquid chromatography (HPLC) according to the national standard GB31645—2018 "Collagen Peptides". The results are as follows: Figure 1 As shown: molecular weight 0-159 Da accounted for 39.07%, 159-389 Da accounted for 42.02%, and 389-2700 Da accounted for 18.91%.

[0066] 5. Preparation and nitrogen content detection of lyophilized brain polypeptide powder

[0067] The brain protein hydrolysate was freeze-dried at -60℃ in a freeze dryer to obtain lyophilized brain polypeptide powder. 0.200g of the lyophilized brain polypeptide powder was taken, and two digestive tablets (4g / tablet) were added. 12mL of concentrated sulfuric acid was slowly added, and the mixture was digested for 3 hours. The nitrogen content was determined using a FOSS 8400 fully automated Kjeldahl nitrogen analyzer. The results are shown in Table 1.

[0068] Table 1 Nitrogen content determination

[0069] Sample Pig brain protein powder Brain protein hydrolysate Brain polypeptide freeze-dried powder Nitrogen content 138.73 mg N / g 8.34 mg N / mL 120.86 mg N / g

[0070] Example 2: Effect of different lipophilic emulsifiers on the encapsulation efficiency of W1 / O / W2 type dual emulsion for encapsulating brain peptide lyophilized powder

[0071] (1) Preparation of inner aqueous phase W1: The lyophilized brain peptide powder was mixed with ultrapure water and vortexed to disperse it evenly. The mixture was then filtered through a 0.22 μm filter membrane to form inner aqueous phase W1. The loading concentration of the lyophilized brain peptide powder in inner aqueous phase W1 was 25 mg / mL.

[0072] (2) Preparation of oil phase O: Add 6% lipophilic emulsifier (polyglycerol ricinoleate (PGPR) or modified soybean lecithin or soybean lecithin or Span 80 or mono- or diglyceride fatty acid ester or lactic acid fatty acid glyceride) to a certain amount of soybean oil, and stir magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, which is oil phase O.

[0073] (3) Preparation of primary emulsion W1 / O: The entire aqueous phase W1 was added dropwise to the entire oil phase O while stirring. The emulsification was carried out by high-energy emulsification at a speed of 15000 rpm for 2 min, and then homogenized by ultrasonication at 450W for 15 min to obtain primary emulsion W1 / O. The volume ratio of oil phase O to aqueous phase W1 was 5:5.

[0074] (4) Preparation of external aqueous phase W2: A certain volume of ultrapure water is preheated in a water bath at 50°C for 15 min, and 4% modified soybean lecithin and 0.1% sucrose fatty acid ester are added to it. Stir at 360 rpm for 30 min to fully dissolve it, which is the external aqueous phase W2.

[0075] (5) Preparation of W1 / O / W2 type double emulsion: The primary emulsion W1 / O and the external aqueous phase W2 (internal-to-external ratio) were mixed at a volume ratio of 5:5, and emulsified by shearing at 10,000 rpm for 2 minutes using a high-energy emulsification method to obtain the double emulsion W1 / O / W2. After storage at room temperature, the W1 / O / W2 type double emulsion containing encapsulated brain peptide lyophilized powder was obtained.

[0076] The encapsulation efficiency (EE%) of the lyophilized brain polypeptide powder in the double emulsion was calculated by the following formula, and the experimental results are shown in Table 2.

[0077]

[0078] C — Concentration of the lyophilized brain polypeptide powder that migrated into the external aqueous phase;

[0079] V – Volume of the supernatant;

[0080] C0 – Concentration of lyophilized brain peptide powder added to the double emulsion;

[0081] —The volume fraction of the internal aqueous phase in the double emulsion;

[0082] V0 – Volume of the double emulsion.

[0083] Table 2. Effect of different types of lipophilic emulsifiers on the encapsulation efficiency of W1 / O / W2 emulsions for encapsulating brain peptide lyophilized powder.

[0084]

[0085] As shown in Table 2, the W1 / O / W2 type double emulsion prepared by PGPR as a lipophilic emulsifier has the highest encapsulation rate.

[0086] Example 3: Effect of PRPR addition amount on the encapsulation efficiency of W1 / O / W2 type dual emulsion for encapsulated brain peptide lyophilized powder

[0087] (1) Preparation of inner aqueous phase W1: The lyophilized brain peptide powder was mixed with ultrapure water and vortexed to disperse it evenly. The mixture was then filtered through a 0.22 μm filter membrane to form inner aqueous phase W1. The loading concentration of the lyophilized brain peptide powder in inner aqueous phase W1 was 25 mg / mL.

[0088] (2) Preparation of oil phase O: Add different proportions of PGPR (1% or 2% or 4% or 6% or 8% or 10%) to a certain amount of soybean oil and stir magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, which is oil phase O.

[0089] (3) Preparation of primary emulsion W1 / O: The aqueous phase W1 prepared in step (1) is added dropwise to the oil phase O prepared in step (2) while stirring. The emulsification is carried out by high-energy emulsification at a speed of 15000 rpm for 2 min, and then the emulsion is homogenized by ultrasonication at 450W for 15 min to obtain primary emulsion W1 / O; the volume ratio of the oil phase O to the aqueous phase W1 is 5:5.

[0090] (4) Preparation of external aqueous phase W2: A certain volume of ultrapure water is preheated in a water bath at 50°C for 15 min, and 4% modified soybean lecithin and 0.1% sucrose fatty acid ester are added to it. Stir at 360 rpm for 30 min to fully dissolve it, which is the external aqueous phase W2.

[0091] (5) Preparation of W1 / O / W2 type double emulsion: The primary emulsion W1 / O and the external aqueous phase W2 were mixed at a volume ratio of 5:5 and emulsified by shearing at 10,000 rpm for 2 minutes using a high-energy emulsification method to obtain the double emulsion W1 / O / W2. After storage at room temperature, the W1 / O / W2 type double emulsion containing the encapsulated brain peptide lyophilized powder was obtained.

[0092] The experimental results are shown in Table 3.

[0093] Table 3. Effect of PGPR addition amount on the encapsulation efficiency of W1 / O / W2 emulsion of encapsulated brain peptide lyophilized powder.

[0094] PGPR addition amount (%) 1 2 4 6 8 10 Embedding rate (%) 49.1 67.88 68.36 69.3 67.47 64.24

[0095] As shown in Table 3, the W1 / O / W2 type double emulsion prepared by using 6% PGPR as a lipophilic emulsifier has the highest encapsulation rate.

[0096] Example 4: Effect of oil-water ratio on the encapsulation efficiency of W1 / O / W2 type dual emulsion for encapsulated brain peptide lyophilized powder

[0097] (1) Preparation of inner aqueous phase W1: The lyophilized brain peptide powder was mixed with ultrapure water and vortexed to disperse it evenly. The mixture was then filtered through a 0.22 μm filter membrane to form inner aqueous phase W1. The loading concentration of the lyophilized brain peptide powder in inner aqueous phase W1 was 25 mg / mL.

[0098] (2) Preparation of oil phase O: Add 6% PGPR to a certain amount of soybean oil and stir magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, which is oil phase O.

[0099] (3) Preparation of primary emulsion W1 / O: The entire aqueous phase W1 was added dropwise to the entire oil phase O while stirring. The emulsification was carried out by high-energy emulsification at a speed of 15000 rpm for 2 min, and then homogenized by ultrasonication at 450W for 15 min to obtain primary emulsion W1 / O. The volume ratio of the oil phase O to the aqueous phase W1 was 5:5, 6:4, 7:3, 8:2, or 9:1.

[0100] (4) Preparation of external aqueous phase W2: A certain volume of ultrapure water is preheated in a water bath at 50°C for 15 min, and 4% modified soybean lecithin and 0.1% sucrose fatty acid ester are added to it. Stir at 360 rpm for 30 min to fully dissolve it, which is the external aqueous phase W2.

[0101] (5) Preparation of W1 / O / W2 type double emulsion: The primary emulsion W1 / O and the external aqueous phase W2 were mixed at a volume ratio of 5:5 and emulsified by high-energy emulsification at 10,000 rpm for 2 min to obtain the double emulsion W1 / O / W2. After storage at room temperature, the W1 / O / W2 type double emulsion containing encapsulated brain peptide lyophilized powder was obtained.

[0102] The experimental results are shown in Table 4.

[0103] Table 4. Effect of oil-water ratio on the encapsulation efficiency of W1 / O / W2 emulsions for encapsulated brain peptide lyophilized powder.

[0104] Oil-water ratio (O:W1) 5:5 6:4 7:3 8:2 9:1 Embedding rate (%) 69.97 72.31 73.14 77.45 74.9

[0105] As shown in Table 4, the W1 / O / W2 type double emulsion prepared with an oil-water ratio of 8:2 has the highest encapsulation rate.

[0106] Example 5: Effect of Hydrophilic Emulsifier Type on Encapsulation Efficiency of W1 / O / W2 Type Dual Emulsion for Encapsulating Brain Peptide Lyophilized Powder

[0107] (1) Preparation of inner aqueous phase W1: The lyophilized brain peptide powder was mixed with ultrapure water and vortexed to disperse it evenly. The mixture was then filtered through a 0.22 μm filter membrane to form inner aqueous phase W1. The loading concentration of the lyophilized brain peptide powder in inner aqueous phase W1 was 25 mg / mL.

[0108] (2) Preparation of oil phase O: Add 6% PGPR to a certain amount of soybean oil and stir magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, which is oil phase O.

[0109] (3) Preparation of the primary emulsion W1 / O: The entire aqueous phase W1 was added dropwise to the entire oil phase O while stirring. The emulsification was carried out by high-energy emulsification at 15000 rpm for 2 min, and then homogenized by ultrasonication at 450 W for 15 min to obtain the primary emulsion W1 / O. The volume ratio of the oil phase O to the aqueous phase W1 was 8:2.

[0110] (4) Preparation of external aqueous phase W2: A certain volume of ultrapure water is preheated in a water bath at 50°C for 15 min. 4% of hydrophilic emulsifier (modified soybean lecithin or soybean phospholipid or Tween 80 or soluble soybean polysaccharide or sodium carboxymethyl cellulose (CMC-Na) or gum arabic) and 0.1% sucrose fatty acid ester are added to it and stirred at 360 rpm for 30 min to fully dissolve it, which is the external aqueous phase W2.

[0111] (5) Preparation of W1 / O / W2 type double emulsion: The primary emulsion W1 / O and the external aqueous phase W2 were mixed at a volume ratio of 5:5 and emulsified by shearing at 10,000 rpm for 2 minutes using a high-energy emulsification method to obtain the double emulsion W1 / O / W2. After storage at room temperature, the W1 / O / W2 type double emulsion containing the encapsulated brain peptide lyophilized powder was obtained.

[0112] The experimental results are shown in Table 5.

[0113] Table 5. Effect of hydrophilic emulsifier type on the encapsulation efficiency of W1 / O / W2 emulsions for encapsulating brain peptide lyophilized powder.

[0114]

[0115] As shown in Table 5, Tween 80, as a hydrophilic emulsifier, produced the W1 / O / W2 type double emulsion with the highest encapsulation rate.

[0116] Example 6: Effect of Tween 80 addition amount on the encapsulation efficiency of W1 / O / W2 type dual emulsion for encapsulated brain peptide lyophilized powder

[0117] (1) Preparation of inner aqueous phase W1: The lyophilized brain peptide powder was mixed with ultrapure water and vortexed to disperse it evenly. The mixture was then filtered through a 0.22 μm filter membrane to form inner aqueous phase W1. The loading concentration of the lyophilized brain peptide powder in inner aqueous phase W1 was 25 mg / mL.

[0118] (2) Preparation of oil phase O: Add 6% PGPR to a certain amount of soybean oil and stir magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, which is oil phase O.

[0119] (3) Preparation of primary emulsion W1 / O: The entire aqueous phase W1 was added dropwise to the entire oil phase O while stirring. The emulsification was carried out by high-energy emulsification at a speed of 15000 rpm for 2 min, and then homogenized by ultrasonication at 450W for 15 min to obtain primary emulsion W1 / O. The volume ratio of oil phase O to aqueous phase W1 was 8:2.

[0120] (4) Preparation of external aqueous phase W2: A certain volume of ultrapure water is preheated in a water bath at 50°C for 15 min. Different proportions of Tween80 (1% or 2% or 4% or 6% or 8% or 10%) and 0.1% sucrose fatty acid ester are added to it and stirred at 360 rpm for 30 min to fully dissolve it, which is the external aqueous phase W2.

[0121] (5) Preparation of W1 / O / W2 type double emulsion: The primary emulsion W1 / O and the external aqueous phase W2 were mixed at a volume ratio of 5:5 and emulsified by shearing at 10,000 rpm for 2 minutes using a high-energy emulsification method to obtain the double emulsion W1 / O / W2. After storage at room temperature, the W1 / O / W2 type double emulsion containing the encapsulated brain peptide lyophilized powder was obtained.

[0122] The experimental results are shown in Table 6.

[0123] Table 6. Effect of Tween 80 addition on the encapsulation efficiency of W1 / O / W2 emulsion for encapsulated brain peptide lyophilized powder.

[0124] Tween 80 addition amount (%) 1 2 4 6 8 10 Embedding rate (%) 32.61 67.02 79.7 66.19 60.68 46.31

[0125] As shown in Table 6, the W1 / O / W2 type double emulsion prepared by using 4% Tween 80 as a hydrophilic emulsifier has the highest encapsulation rate.

[0126] Example 7: Effect of internal and external comparison on the encapsulation efficiency of W1 / O / W2 type dual emulsion for encapsulated brain peptide lyophilized powder

[0127] (1) Preparation of inner aqueous phase W1: The lyophilized brain peptide powder was mixed with ultrapure water and vortexed to disperse it evenly. The mixture was then filtered through a 0.22 μm filter membrane to form inner aqueous phase W1. The loading concentration of the lyophilized brain peptide powder in inner aqueous phase W1 was 25 mg / mL.

[0128] (2) Preparation of oil phase O: Add 6% PGPR to a certain amount of soybean oil and stir magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, which is oil phase O.

[0129] (3) Preparation of the primary emulsion W1 / O: The entire aqueous phase W1 was added dropwise to the entire oil phase O while stirring. The emulsification was carried out by high-energy emulsification at 15000 rpm for 2 min, and then homogenized by ultrasonication at 450 W for 15 min to obtain the primary emulsion W1 / O. The volume ratio of the oil phase O to the aqueous phase W1 was 8:2.

[0130] (4) Preparation of external aqueous phase W2: A certain volume of ultrapure water is preheated in a water bath at 50°C for 15 min. 4% of hydrophilic emulsifier Tween 80 and 0.1% sucrose fatty acid ester are added to it and stirred at 360 rpm for 30 min to fully dissolve it, which is the external aqueous phase W2.

[0131] (5) Preparation of W1 / O / W2 type dual emulsion: The primary emulsion W1 / O and the external aqueous phase W2 are mixed at a certain volume ratio (5:5, 4:6, 3:7, 2:8, or 1:9), and emulsified by high-energy emulsification at 10,000 rpm for 2 min to obtain the dual emulsion W1 / O / W2. After storage at room temperature, the W1 / O / W2 type dual emulsion containing encapsulated brain polypeptide lyophilized powder is obtained.

[0132] The experimental results are shown in Table 7.

[0133] Table 7. Effect of internal and external comparison on the encapsulation efficiency of W1 / O / W2 emulsions for encapsulated brain peptide lyophilized powder.

[0134] Inner and outer ratio 5:5 4:6 3:7 2:8 1:9 Embedding rate (%) 79.69 80.77 76.2 61.13 57.82

[0135] As shown in Table 7, the W1 / O / W2 type double emulsion with an internal-to-external ratio of 4:6 has the highest encapsulation efficiency.

[0136] Example 8: Preparation of a Highly Stable W1 / O / W2 Type Dual Emulsion

[0137] (1) Preparation of inner aqueous phase W1: The lyophilized brain peptide powder was mixed with ultrapure water and vortexed to disperse it evenly. The mixture was then filtered through a 0.22 μm filter membrane to form inner aqueous phase W1. The loading concentration of the lyophilized brain peptide powder in inner aqueous phase W1 was 25 mg / mL.

[0138] (2) Preparation of oil phase O: Add 6% PGPR to a certain amount of soybean oil and stir magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, which is oil phase O.

[0139] (3) Preparation of the primary emulsion W1 / O: The entire aqueous phase W1 was added dropwise to the entire oil phase O while stirring. The emulsification was carried out by high-energy emulsification at 15000 rpm for 2 min, and then homogenized by ultrasonication at 450 W for 15 min to obtain the primary emulsion W1 / O. The volume ratio of the oil phase O to the aqueous phase W1 was 8:2.

[0140] (4) Preparation of external aqueous phase W2: A certain volume of ultrapure water is preheated in a water bath at 50°C for 15 min. 4% Tween 80, 0.1% sucrose fatty acid ester and xanthan gum in different proportions (0.2% or 0.4% or 0.6% or 0.8% or 1.0%) are added to it and stirred at 360 rpm for 30 min to fully dissolve it, which is the external aqueous phase W2.

[0141] (5) Preparation of highly stable W1 / O / W2 type dual emulsion: The primary emulsion W1 / O and the external aqueous phase W2 were mixed at a volume ratio of 4:6, and emulsified by high-energy emulsification at 10,000 rpm for 2 min to obtain the dual emulsion W1 / O / W2. After storage at room temperature, the highly stable W1 / O / W2 type dual emulsion encapsulating brain peptide lyophilized powder was obtained.

[0142] Example 9: Evaluation of the high stability of W1 / O / W2 type dual emulsion for encapsulated brain peptide lyophilized powder

[0143] 1. Observation of the apparent morphology of the emulsion

[0144] Take the W1 / O / W2 type dual emulsions with different xanthan gum additions prepared in Example 8, add them to a 20mL vial, seal, cap, and let stand at room temperature for 30 days to observe the changes in the appearance of the emulsion. Figure 2 As shown: Freshly prepared emulsions are uniform and fine, free of impurities and flocculation, and do not separate into layers. After standing at room temperature for 30 days, the emulsion without xanthan gum showed obvious separation, the emulsion with 0.2% xanthan gum showed slight separation at the bottom, and the other emulsions showed no obvious changes.

[0145] 2. Observation of droplet morphology

[0146] The highly stable W1 / O / W2 dual emulsion was observed using an optical microscope. 2.5 μL of each sample was placed on a slide and diluted 5-fold directly onto the microscope slide with ultrapure water. A coverslip was then gently placed over the emulsion. The W1 / O / W2 emulsion was observed using a 40x objective lens. Results are as follows: Figure 3 As shown, the emulsion without xanthan gum has a larger particle size, which is detrimental to emulsion stability, while the emulsion with xanthan gum has a smaller particle size. Among them, the emulsion with 0.8% xanthan gum has the smallest particle size and more double-encapsulated structures, making it the optimal choice.

[0147] 3. Determination of the encapsulation efficiency of lyophilized brain polypeptide powder

[0148] Take 1 mL of each of the W1 / O / W2 double emulsions prepared in Example 8 with different xanthan gum additions, centrifuge at 1800 rpm for 10 min, collect the supernatant, centrifuge at 10000 rpm for 30 min, collect the supernatant, dilute 100 times with 0.15 M NaCl solution, and measure the absorbance at 268 nm to calculate the concentration of lyophilized brain peptide powder in the supernatant. Prepare a standard curve using lyophilized brain peptide powder of known concentration. The encapsulation efficiency (EE) of lyophilized brain peptide powder in the double emulsion is calculated by the following formula.

[0149]

[0150] C — Concentration of the lyophilized brain polypeptide powder that migrated into the external aqueous phase;

[0151] V – Volume of the supernatant;

[0152] C0 – Concentration of lyophilized brain peptide powder added to the double emulsion;

[0153] —The volume fraction of the internal aqueous phase in the double emulsion;

[0154] V0 – Volume of the double emulsion.

[0155] The results are as follows Figure 4 As shown, from Figure 4 It can be seen that the emulsion encapsulation rate is high for different amounts of xanthan gum, indicating that xanthan gum helps stabilize the W1 / O / W2 dual emulsion system. Among them, the encapsulation rate is the highest when 0.8% xanthan gum is added.

[0156] 4. Measurement of emulsifying activity (EAI) and emulsifying stability (ESI)

[0157] Take 20 μL of each of the emulsions prepared in Example 8 with different xanthan gum additions, and add them to 4 mL of 0.1% sodium dodecyl sulfate (SDS) solution and mix thoroughly. Measure the absorbance A0 at 500 nm, and after standing for 60 min, measure the absorbance A10. 60 The emulsion activity index (EAI) and emulsion stability index (ESI) are calculated using the following formulas.

[0158]

[0159] In the formula: A0 is the absorbance value at 0 min;

[0160] N is the dilution factor;

[0161] C represents the sample concentration, in g / mL;

[0162] θ is the volume of the oil phase;

[0163] ΔT is the time difference, in min;

[0164] ΔA represents the difference in absorbance within ΔT.

[0165] The results are as follows Figure 5 As shown, the EAI and ESI of the emulsion with added xanthan gum were significantly higher than those without. The emulsifying properties of proteins depend on their ability to reduce interfacial tension between two phases. With increasing xanthan gum content, the trends in EAI and ESI were roughly similar, with the highest EAI and ESI (15.282 m) observed when the xanthan gum content was 0.8%. 2 / g and 452.2min, which demonstrate superior interfacial properties and stability compared to other xanthan gum additions.

[0166] 5. Storage stability determination

[0167] Double emulsions may separate into layers during storage, with the upper layer being the emulsion retention layer and the lower layer being a clear or cloudy layer. Freshly prepared double emulsions were bottled and stored at room temperature for 0, 0.5, 1, 3, 7, 14, and 30 days, and the emulsion separation was observed. The results are as follows: Figure 6 As shown. Its stability index calculation formula is:

[0168]

[0169] Where: TSI—Emulsion stability index, %

[0170] H0 — Height of the emulsion sample, in cm

[0171] H1 — Height of the lower layer of the emulsion after separation, in cm

[0172] After 30 days of observation at room temperature, it was found that the emulsion without xanthan gum separated into layers after 12 hours. From day 0 to day 7, the double emulsions with 0.2% to 1.0% xanthan gum remained in good condition without separation. On day 30, it was found that, except for the double emulsion with 0.2% xanthan gum, which showed slight separation, the other emulsions remained in good condition without separation.

[0173] 6. Determination of centrifugal retention rate

[0174] Centrifugal retention rate is an important indicator for evaluating the stability of double emulsions. The method for determining the centrifugal retention rate of double emulsions involves centrifugation, where the prepared double emulsion is separated into layers under centrifugal force. The calculated emulsion layer retention rate serves as a measure of the centrifugal stability of the double emulsion. 1 mL of freshly prepared double emulsion is placed in a 1.5 mL centrifuge tube and centrifuged at 2000 rpm for 10 min. The double emulsion separates into layers, with an aqueous phase precipitating at the bottom. The total volume of the emulsion and the volume of the precipitated aqueous phase in the centrifuge tube are measured. The results are as follows: Figure 7 As shown. The centrifugal retention rate of the W1 / O / W2 type double emulsion was calculated according to the formula.

[0175]

[0176] In the formula, V t V represents the total volume of the liquid layer in a centrifuge tube. c This indicates the volume of the aqueous phase that has separated after stratification.

[0177] The emulsion without xanthan gum showed a centrifugal retention rate of only 50.2%, indicating that the emulsion system without xanthan gum is unstable and prone to stratification. This is consistent with the results of static observation and storage stability. The emulsion with 0.2% xanthan gum showed a centrifugal retention rate of 53.5%, which is not significantly different from the emulsion without xanthan gum. The emulsions with other xanthan gum additions showed a centrifugal retention rate of 100%, indicating good stability.

[0178] Example 10: In vitro evaluation of W1 / O / W2 type dual emulsions containing encapsulated brain peptide lyophilized powder

[0179] In vitro stability of brain polypeptide lyophilized powder under simulated gastrointestinal fluid conditions.

[0180] (1) Simulated gastric juice (SGF) digestion: The simulated gastric juice consisted of 3.2 mg / mL pepsin, 2.0 mg / mL NaCl, and 0.7% (v / v) HCl, with a pH of 1.2. 1 mL of the sample was mixed with 3 mL of SGF, and the pH was adjusted to 2.0 using 1 M NaOH. The temperature was maintained at 37°C, and the magnetic stirring speed was 100 rpm for 2 hours. The pH of the 4 mL of chyme produced during the simulated gastric juice digestion stage was adjusted to 7.0.

[0181] (2) Simulated Intestinal Fluid (SIF) Digestion: The simulated intestinal fluid contained 1.5 mL of salt solution (36.7 mg / mL CaCl2 and 218.7 mg / mL NaCl), 2.5 mL of enzyme solution (24 mg / mL lipase and 24 mg / mL pancreatin), and 3.5 mL of bile salt solution (54 mg / mL). The enzyme and bile salt were dissolved in phosphate buffer solution (pH 7.0). 1 mL of SIF was added to the pH-adjusted chyme, and the pH of the system was monitored. The pH was maintained at 7.0 using 0.1 M NaOH solution, and digestion was continued at 37°C with magnetic stirring at 100 rpm for 2 hours. The volume of NaOH consumed was recorded. The percentage of free fatty acids (FFA) released was calculated using the following formula:

[0182] %FFA = (V NaOH ×C NaOH ×M Lipid ) / (2×W Liqid )×100

[0183] In the formula, V NaOH The volume of NaOH consumed (mL), C NaOH M represents the molar concentration of NaOH (0.1M). Lipid W represents the average molecular weight of soybean oil. Lipid The initial mass (g) of soybean oil in the emulsion.

[0184] After passing through simulated gastric juice, the unencapsulated lyophilized brain peptide powder lost its activity due to pepsin and low pH conditions, while the brain peptide lyophilized powder encapsulated in a double emulsion retained its activity. After the emulsion enters the small intestine for digestion, the oil phase is decomposed into glycerol and FFA under the action of lipase. The FFA release rate of the W1 / O / W2 emulsion initially increases and then levels off during digestion, as shown in the results below. Figure 8 After 120 minutes of digestion, the free fatty acid (FFA) release rate of the hydrolysate from the unencapsulated brain peptide lyophilized powder, simulating intestinal digestion, was only 5.44%. The FFA release rate of the W1 / O / W2 emulsion without xanthan gum was 63.03%. The FFA release rate of the dual emulsion with xanthan gum reached 80%–90%, with the W1 / O / W2 emulsion with 0.8% xanthan gum showing the highest FFA release rate of 91.5%. This indicates that the brain peptide lyophilized powder encapsulated in the dual emulsion can stably reach the intestines after passing through gastric juices and be efficiently released in the intestines, with a release rate 45.17% higher than that of the W1 / O / W2 emulsion without xanthan gum.

[0185] (3) Chemical stability (S) * ) and biological accessibility (B * The final chyme remaining after small intestinal digestion was centrifuged at 10000×g for 30 min and separated into three layers: a bottom precipitate phase, a middle micelle phase, and an upper oil phase. The chemical stability (S) of the lyophilized brain polypeptide powder was also assessed. * ) and biological accessibility (B * The measurement is performed according to the following formula. The result is as follows: Figure 9 As shown.

[0186] %S*=C Digesta / C Initial ×100

[0187] %B*=C Micelle / C Initial ×100

[0188] Where C Initial This is the initial concentration of the lyophilized brain peptide powder in the emulsion, C. Digesta It is the concentration of lyophilized brain polypeptide powder in the total chyme collected after the small intestine stage, C Micelle It is the concentration of the lyophilized brain polypeptide powder in the mixed micelle phase.

[0189] Studies simulating gastrointestinal digestion showed that W1 / O / W2 type double emulsions with different xanthan gum additions exhibited improved chemical stability and bioavailability compared to unencapsulated hydrolysates. Specifically, the 0.8% xanthan gum sample showed a 39.03% improvement in chemical stability and an 86.60% improvement in bioavailability compared to the hydrolysate. These results indicate that encapsulating lyophilized brain peptides in W1 / O / W2 type double emulsions can prevent their release in the stomach. The reduced bitterness and improved gastrointestinal digestibility are attributed to the peptides being dispersed in the aqueous phase, with a lipid layer surrounding the water droplets. W1 / O / W2 type double emulsions effectively mask bitterness and improve gastrointestinal stability, potentially expanding the application of bioactive ingredients in food.

[0190] In summary, a highly stable W1 / O / W2 type dual emulsion was prepared by adding 0.8% xanthan gum.

[0191] Example 11 Preparation of Fruit-Flavored Brain Polypeptide Oral Liquid

[0192] (1) Take 20 mL of the high-stability W1 / O / W2 type double emulsion with xanthan gum added at 0.8% in Example 8 and adjust the pH to 7.0;

[0193] (2) Add 4% peach concentrate and 0.1% lemon concentrate, stir well, adjust the pH to 7.0, and obtain the prepared mixture;

[0194] (3) Filter the prepared mixture through a 500-mesh filter cloth, and then fill it to obtain an oral liquid;

[0195] (4) The oral liquid filled in step (3) is sterilized by high temperature and high pressure. The sterilization method is: high pressure steam sterilization at 121℃ for 15 minutes, and then cooled to a core temperature of less than 35℃ to obtain the finished product.

[0196] (5) Sensory evaluation was conducted on the brain polypeptide oral liquid using a sensory evaluation scoring method. Four indicators were selected: odor and taste, color, texture and impurities. Ten trained students were asked to conduct sensory evaluation of the obtained fruit-flavored brain polypeptide oral liquid. The evaluation criteria are shown in Table 8, and the evaluation results are shown in Table 9.

[0197] The difference between Example 12 and Example 11 is limited to the fact that in step (2), the mango concentrate is 3% and the orange concentrate is 0.5%.

[0198] The difference between Example 13 and Example 11 is limited to the fact that in step (2), apple concentrate is 4% and peach concentrate is 1%.

[0199] The difference between Example 14 and Example 11 is limited to the 3% grape concentrate in step (2).

[0200] Table 8 Flavor Evaluation Criteria for Brain Polypeptide Oral Liquid

[0201]

[0202] Table 9 Sensory evaluation results of brain polypeptide oral liquid

[0203]

[0204] Sensory evaluation results show that the lyophilized brain polypeptide powder aqueous solution exhibits a bitter taste due to the exposure of hydrophobic amino acids during the hydrolysis of porcine brain protein powder, which directly bind to bitter taste receptors in the oral cavity, resulting in a significant unpleasant odor and making it difficult to swallow. Furthermore, a small amount of microparticles from the lyophilized brain polypeptide powder aqueous solution that permeated the filter membrane swelled and expanded in the water, transforming into visible floating matter, leading to a foreign body sensation and resulting in the lowest overall score of only 59 points. The W1 / O / W2 type double emulsion (Example 8) effectively masked this unpleasant odor, but the use of emulsifiers introduced a slight peculiar odor. To reduce the impact of the unpleasant flavor of the emulsifiers and enrich the flavor of the brain polypeptide oral liquid, this invention optimizes the flavor by adding concentrated fruit juice. Sensory evaluation results show that the various fruit-flavored brain polypeptide oral liquids (Examples 11-14) have a better taste, strong palatability, a sweet and sour flavor that is not cloying, significantly masking unpleasant flavors, improving the sensory quality of the beverage, and increasing consumer acceptance.

Claims

1. A method for preparing a highly stable W1 / O / W2 type dual emulsion for encapsulating lyophilized brain polypeptide powder, characterized in that, Includes the following steps: S1: Preparation of inner aqueous phase W1: After mixing the lyophilized brain peptide powder with ultrapure water, the mixture is vortexed to disperse it evenly, and then filtered through a 0.22μm filter membrane to form inner aqueous phase W1, wherein the loading of the lyophilized brain peptide powder is 25mg / mL. S2: Preparation of oil phase O: Dissolve a lipophilic emulsifier in edible oil and stir magnetically to form oil phase O; S3: Preparation of water-in-oil primary emulsion W1 / O: The oil phase O and the inner aqueous phase W1 are initially mixed, and then homogenized by stirring, homogenizing, and ultrasonication to obtain a water-in-oil primary emulsion W1 / O; the mass ratio of oil phase O to inner aqueous phase W1 is 8:2; the homogenization conditions in step S3 are 10000-15000 rpm shearing for 2 min, and the ultrasonic homogenization conditions are 450W ultrasonication for 15 min; S4: Preparation of external aqueous phase W2: Add 4% Tween 80 hydrophilic emulsifier, 0.1% sucrose fatty acid ester and 0.8% xanthan gum to ultrapure water at 50℃, preheat for 15 minutes, stir at 360 rpm for 30 minutes to dissolve, and the external aqueous phase W2 is obtained. S5: Preparation of high-stability W1 / O / W2 type dual emulsion: The primary emulsion W1 / O and the external aqueous phase W2 are mixed in a volume ratio of 4:6, and emulsified by high-energy emulsification method at 10000 rpm for 2 min to obtain the high-stability W1 / O / W2 type dual emulsion for encapsulating brain peptide lyophilized powder. In step S2, the edible oil is soybean oil, the lipophilic emulsifier is polyglycerol ricinoleate (PGPR), and the amount of lipophilic emulsifier added is 6%.

2. The application of the emulsion prepared by the method of claim 1 in the preparation of fruit-flavored brain polypeptide oral liquid.

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