Preparation method and application of W1 / O / W2 type dual emulsion based on embedded brain polypeptide freeze-dried powder

Abstract

The invention discloses a preparation method and application of a W1 / O / W2 type dual emulsion based on embedded 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 internal water phase, mixing the oil phase and the internal water phase, and homogenizing to obtain W1 / O primary emulsion; and taking a solution loaded with Tween 80, xanthan gum and sucrose fatty acid ester as an external water phase, mixing the external water phase with the W1 / O primary emulsion, performing homogenization treatment, adding fruit juice concentrated juice, and performing stirring, canning and sterilization to obtain the fruity brain polypeptide oral liquid. The double emulsion provided by the invention can significantly improve the release and absorption of brain polypeptide in intestinal tracts, improves the oral bioavailability, and can be used for small intestine targeted delivery of hydrophilic bioactive substances. The obtained emulsion has good stability after being placed at normal temperature. The obtained fruity brain polypeptide oral liquid is high in palatability, and the acceptability of consumers is greatly improved.

Description

Technical Field

[0001] The invention 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 an application thereof. Background Art

[0002] Brain peptides are mainly obtained from pig brain by enzymatic hydrolysis and quantitative ultrafiltration, and a mixture of 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 peptides have the functions of regulating and improving neuronal metabolism, activating brain neuron regeneration, promoting synapse formation, preventing neuronal apoptosis, improving brain functions related to cognition, language, movement, etc., and protecting nerve cells from various ischemic and neurotoxin damage. In the process of enzymatic hydrolysis of proteins to form peptides, hydrophobic amino acids are often exposed, causing them to directly bind to bitter taste receptors in the mouth, resulting in bitter taste. Due to its poor sensory properties and easy degradation under gastrointestinal conditions, its wide application as a functional ingredient is subject to certain restrictions. At the same time, this uncomfortable sensory experience reduces consumers' acceptance of peptide foods.

[0003] Chinese patent CN103445263A discloses an enzymatic ovalbumin compound nutritious oral liquid, wherein egg white is enzymatically hydrolyzed to obtain small molecule polypeptides with biological activities such as anti-oxidation, anti-fatigue, and blood pressure reduction, and then wolfberry extract is added to prepare an enzymatic ovalbumin compound oral nutritious liquid that is rich in nutrition, enhances the body's immunity, and is suitable for the taste of adults. In the prior art, enzymatic albumin is mainly compounded with plant extracts, which mainly achieves the effects of anti-oxidation and enhancing immunity, but there are often problems of poor product stability and palatability during the processing.

[0004] In order to maintain the biological activity of brain polypeptides and the stability of small molecule polypeptides during storage, processing and ingestion, and ultimately obtain products that meet the taste of consumers, the present invention adopts a method of constructing a water-in-oil-in-water (W1 / O / W2) double emulsion to mask the bitterness of brain polypeptides, improve their unpleasant taste, and enhance consumer acceptability.

[0005] Chinese patent CN 117838633 B discloses a preparation method and application of a W1 / O / W2 type double emulsion based on embedding IgY, wherein a mixed solution of butter and corn oil loaded with polyglycerol ricinoleate (PGPR) is used as the oil phase, and a solution loaded with IgY is used as the internal aqueous phase. The oil phase is then mixed with the internal aqueous phase and subjected to shear homogenization to obtain a W1 / O emulsion. The polysaccharide dispersion as the external aqueous phase is mixed with the W1 / O emulsion and subjected to shear homogenization, and stored at low temperature to obtain a double emulsion co-embedded with IgY, which can improve the intestinal adhesion of IgY and the survival rate in the gastrointestinal tract. However, the double emulsion prepared by this method has an IgY release rate of only 40% when passing through simulated intestinal fluid, and the release rate needs to be further improved to facilitate the absorption of nutrients.

[0006] Double emulsion carriers have the characteristics of flavor masking, controlling the release of active ingredients, and improving stability. However, in actual production, the emulsifier used in the double emulsion will produce certain unpleasant flavors, which have a certain impact on the taste of the product. Oral liquid products not only need to have high nutritional value, but also need to have good sensory quality and palatability. Therefore, the present invention also optimizes the flavor of the W1 / O / W2 double emulsion that encapsulates brain polypeptide freeze-dried powder to solve the problems of poor product palatability and single taste. The present invention further adds fruit juice concentrate to the W1 / O / W2 double emulsion to significantly mask the unpleasant flavor and improve the sensory quality of the beverage. The resulting emulsion is stored at room temperature for 3 months without stratification. 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 double emulsion using brain polypeptide freeze-dried 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] The invention provides a method for preparing freeze-dried brain polypeptide powder.

[0009] The brain protein hydrolysate is placed in a freeze dryer for vacuum freeze drying to obtain brain polypeptide freeze-dried powder.

[0010] It is further specified that the preparation method of the brain polypeptide freeze-dried powder is as follows: after dissolving 0.5g of pig brain protein powder in 15mL of ultrapure water, adding 0.1g of trypsin for enzymatic hydrolysis for 2h, inactivating the enzyme in a boiling water bath for 10min, adjusting the pH to 7.0, and filtering to obtain brain protein hydrolyzate.

[0011] It is further defined that the method for obtaining the pig brain protein powder is as follows:

[0012] Thaw the frozen pig brain (without fascia) and crush it with a wall breaker to obtain pig brain homogenate. Mix the pig brain homogenate with edible alcohol in a mass ratio of 1:2, stir and extract in a 78℃ water bath for 2h, let stand for 10min, and filter the extract to obtain a filter cake. Repeat the above operation for 5 times for a total of 10h, and place the final filter cake in a 45℃ vacuum drying oven for 12h to obtain pig brain protein powder.

[0013] The present invention develops a novel emulsion system for encapsulating freeze-dried brain polypeptide powder, which solves the problem that freeze-dried brain polypeptide powder is easily inactivated in the gastrointestinal tract, improves the release rate and storage stability of freeze-dried brain polypeptide powder in the intestine, and improves the bitterness and fishy smell of the enzymatic hydrolyzate.

[0014] The first aspect of the present invention is to provide a method for preparing a high-stability W1 / O / W2 double emulsion for embedding freeze-dried brain polypeptide powder, comprising the following steps:

[0015] S1: Preparation of inner aqueous phase W1: After mixing the brain polypeptide freeze-dried powder with ultrapure water, vortex to make it evenly dispersed, and filter through a 0.22 μm filter membrane to form the inner aqueous phase W1;

[0016] S2: Preparation of oil phase O: dissolving the lipophilic emulsifier in edible oil and forming oil phase O through magnetic stirring;

[0017] S3: Preparation of colostrum W1 / O: Preliminarily mix the oil phase O and the internal water phase W1, and then stir, homogenize with a homogenizer and homogenize with ultrasound to obtain water-in-oil colostrum W1 / O;

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

[0019] S5: Preparation of W1 / O / W2 type double emulsion: adding the colostrum W1 / O dropwise into the external aqueous phase W2, and obtaining the W1 / O / W2 type double emulsion encapsulating the freeze-dried brain polypeptide powder through stirring, high-speed shearing, homogenization and ultrasonic emulsification;

[0020] S6: Preparation of a high-stability W1 / O / W2 double emulsion: A stabilizer is added to the W1 / O / W2 double emulsion, and after stirring, homogenizing, and ultrasonic emulsification, a high-stability W1 / O / W2 double emulsion can be obtained.

[0021] In a specific embodiment, the loading amount 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 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 di-glycerol fatty acid esters, soybean lecithin, modified soybean lecithin, lactic acid fatty acid glyceride and other lipophilic emulsifiers; preferably polyglycerol ricinoleate (PGPR).

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

[0025] In another specific embodiment, in step S3, the inner water phase W1 is dripped into the oil phase O while stirring, and the volume ratio of the oil phase O to the inner water phase W1 is one of 5:5, 6:4, 7:3, 8:2, and 9:1.

[0026] In another specific embodiment, in S4, W2 is prepared by adding a hydrophilic emulsifier and sucrose fatty acid ester into deionized water and stirring to fully dissolve, wherein 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 contained in W2 in step S4 is 1-10%, preferably 4%. The concentration of the sucrose fatty acid ester is 0.1%. The dissolution is performed by stirring at 50° C. for 30 min.

[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, 9:1; preferably 6:4.

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

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

[0031] The second aspect of the present invention also provides a high-stability W1 / O / W2 double emulsion based on embedded brain polypeptide freeze-dried powder, characterized in that it is prepared by any one of the methods described in the first aspect of the present invention.

[0032] The third aspect of the present invention is to provide the use of the high-stability W1 / O / W2 double emulsion based on embedded brain polypeptide freeze-dried powder described in the second aspect in the preparation of fruity brain polypeptide oral liquid; the preparation steps are as follows:

[0033] a. The pH of the high stability W1 / O / W2 double emulsion is adjusted to 7.0;

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

[0035] c. The emulsion obtained in step b is filtered through a 500-mesh filter cloth and then filled;

[0036] d. The emulsion obtained in step c is sterilized by high temperature and high pressure sterilization: high pressure steam sterilization at 121 ° C for 15 min, cooled to a center temperature of less than 35 ° C, and the fruity brain polypeptide oral solution is obtained;

[0037] e. Conduct sensory evaluation on the above-mentioned fruity brain polypeptide oral liquid.

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

[0039] The present invention is beneficial in that:

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

[0041] (2) A W1 / O / W2 system with high embedding rate and high stability suitable for embedding brain polypeptide freeze-dried powder was successfully established. The formula and preparation method of the present invention, by compounding Tween 80 and xanthan gum in a specific ratio, helps to improve the stability of the W1 / O / W2 emulsion, and it still does not stratify after being placed at room temperature for 3 months.

[0042] (3) This study used a high-stability W1 / O / W2 double emulsion to develop a fruit-flavored peptide beverage to mask its unpleasant flavor. The product had better sensory quality and stronger palatability, which improved consumer acceptance.

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

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

[0045] (6) The oral liquid of the present invention is more convenient to carry and easy to absorb after being filled, and high-pressure sterilization can extend the shelf life, making the oral liquid more hygienic and safe. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 This is the HPLC chart of brain protein hydrolysate.

[0047] Figure 2 This is the apparent diagram of the W1 / O / W2 double emulsion prepared in Example 8. The amounts of xanthan gum added are 0%, 0.2%, 0.4%, 0.6%, 0.8% and 1.0% from left to right.

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

[0049] Figure 4 It is the embedding efficiency of the W1 / O / W2 double emulsion prepared in Example 8.

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

[0051] Figure 6 It is the storage TSI value of the W1 / O / W2 double emulsion prepared in Example 8.

[0052] Figure 7 It is the centrifugal retention rate of the W1 / O / W2 double emulsion prepared in Example 8.

[0053] Figure 8 The free fatty acid (FFA) release rate of the W1 / O / W2 double emulsion prepared in Example 8 after being subjected to simulated gastrointestinal fluid in vitro.

[0054] Fig. 9 The chemical stability (S) of the W1 / O / W2 double emulsion prepared in Example 8 after being subjected to simulated gastrointestinal fluid in vitro * ) and bioaccessibility (B * ). DETAILED DESCRIPTION

[0055] The purposes and functions of the present invention and methods for achieving these purposes 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 different forms. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

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

[0057] 1. Preparation of pig brain protein powder

[0058] Thaw the frozen pig brain (without fascia) and crush it with a wall breaker to obtain a homogenate. Mix the pig brain homogenate with edible alcohol in a ratio of 1:2, stir and extract in a 78℃ water bath for 2h, let stand for 10min, and filter the extract to obtain a filter cake. Repeat the above operation for 5 times for a total of 10h, and place the final filter cake in a 45℃ vacuum drying oven to dry for 12h to obtain pig brain protein powder.

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

[0060] Take 0.200 g of dried pig brain protein powder, add two digestion tablets (4 g / tablet), slowly add 12 mL of concentrated sulfuric acid, digest for 3 h, and determine the nitrogen content on 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 pig brain protein powder in 15mL of ultrapure water, add 0.3g of trypsin and hydrolyze at 50℃ for 1h. Inactivate the enzyme in boiling water bath for 10min, adjust the pH to 7.0, filter to remove the residue, and collect the filtrate to obtain the brain protein hydrolyzate.

[0063] 4. Detection of nitrogen content and molecular weight of 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 measure the nitrogen content using a FOSS8400 fully automatic Kjeldahl nitrogen analyzer. The results are shown in Table 1.

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

[0066] 5. Preparation of Brain Peptide Lyophilized Powder and Detection of Nitrogen Content

[0067] The brain protein hydrolysate was placed in a freeze dryer and vacuum-freeze dried at -60°C to obtain brain polypeptide freeze-dried powder. 0.200 g of brain polypeptide freeze-dried powder was added with two digestion tablets (4 g / tablet), and 12 mL of concentrated sulfuric acid was slowly added. The mixture was digested for 3 h, and the nitrogen content was determined by a FOSS 8400 fully automatic 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 138.73mgN / g 8.34mgN / mL 120.86mgN / g

[0070] Example 2 Effect of different lipophilic emulsifiers on the embedding efficiency of W1 / O / W2 double emulsions for embedding brain polypeptide freeze-dried powder

[0071] (1) Preparation of internal aqueous phase W1: After the brain polypeptide freeze-dried powder is mixed with ultrapure water, vortexed to make it evenly dispersed, and filtered through a 0.22 μm filter membrane to form an internal aqueous phase W1. The loading concentration of the brain polypeptide freeze-dried powder in the internal aqueous phase W1 is 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- and di-glycerol fatty acid esters 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, i.e., oil phase O.

[0073] (3) Preparation of colostrum W1 / O: The entire internal water phase W1 is added dropwise into the entire oil phase O while stirring, and emulsified by high-energy emulsification at a speed of 15,000 rpm for 2 minutes, followed by ultrasonic homogenization at 450 W for 15 minutes to obtain colostrum W1 / O; the volume ratio of the oil phase O to the internal water phase W1 is 5:5.

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

[0075] (5) Preparation of W1 / O / W2 type double emulsion: Colostrum W1 / O and external aqueous phase W2 (internal and external ratio) were mixed at a volume ratio of 5:5, and emulsified by high-energy emulsification method at a rotation speed of 10,000 rpm for 2 minutes to obtain double emulsion W1 / O / W2. The mixture was stored at room temperature to obtain W1 / O / W2 type double emulsion containing brain polypeptide freeze-dried powder.

[0076] The embedding efficiency EE (%) of brain polypeptide freeze-dried powder in double emulsion was calculated by the following formula. The experimental results are shown in Table 2.

[0077]

[0078] C——Concentration of brain polypeptide freeze-dried powder that migrated to the external aqueous phase;

[0079] V——the volume of the supernatant;

[0080] C0——the concentration of brain polypeptide freeze-dried powder added to the double emulsion;

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

[0082] V0 – volume of double emulsion.

[0083] Table 2 Effect of different lipophilic emulsifier types on the embedding efficiency of W1 / O / W2 emulsions for embedding brain polypeptide freeze-dried powder

[0084]

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

[0086] Example 3 Effect of PRPR Addition Amount on the Embedding Rate of W1 / O / W2 Double Emulsion Encapsulating Brain Polypeptide Lyophilized Powder

[0087] (1) Preparation of internal aqueous phase W1: After the brain polypeptide freeze-dried powder is mixed with ultrapure water, vortexed to make it evenly dispersed, and filtered through a 0.22 μm filter membrane to form an internal aqueous phase W1. The loading concentration of the brain polypeptide freeze-dried powder in the internal aqueous phase W1 is 25 mg / mL.

[0088] (2) Preparation of oil phase O: Different proportions of PGPR (1%, 2%, 4%, 6%, 8%, or 10%) were added to a certain amount of soybean oil, and the mixture was stirred magnetically at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, i.e., oil phase O.

[0089] (3) Preparation of colostrum W1 / O: The internal water phase W1 prepared in step (1) is dripped into the oil phase O prepared in step (2) while stirring, and emulsified by high-energy emulsification method at a rotation speed of 15,000 rpm for 2 minutes, and then ultrasonically homogenized at 450W for 15 minutes to obtain colostrum W1 / O; the volume ratio of the oil phase O to the internal water phase W1 is 5:5.

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

[0091] (5) Preparation of W1 / O / W2 double emulsion: Colostrum W1 / O and external water phase W2 were mixed at a volume ratio of 5:5, and emulsified by high-energy emulsification method at a rotation speed of 10,000 rpm for 2 minutes to obtain double emulsion W1 / O / W2. The mixture was stored at room temperature to obtain W1 / O / W2 double emulsion containing brain polypeptide freeze-dried powder.

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

[0093] Table 3 Effect of PGPR addition on the embedding efficiency of W1 / O / W2 emulsions encapsulating brain polypeptide freeze-dried 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 double emulsion prepared with 6% PGPR as the lipophilic emulsifier has the highest embedding rate.

[0096] Example 4 Effect of Oil-Water Ratio on the Embedding Efficiency of W1 / O / W2 Double Emulsion for Embedding Brain Polypeptide Lyophilized Powder

[0097] (1) Preparation of internal aqueous phase W1: After the brain polypeptide freeze-dried powder is mixed with ultrapure water, vortexed to make it evenly dispersed, and filtered through a 0.22 μm filter membrane to form an internal aqueous phase W1. The loading concentration of the brain polypeptide freeze-dried powder in the internal aqueous phase W1 is 25 mg / mL.

[0098] (2) Preparation of oil phase O: 6% PGPR was added to a certain amount of soybean oil and magnetically stirred at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, i.e., oil phase O.

[0099] (3) Preparation of colostrum W1 / O: The entire internal aqueous phase W1 is added dropwise into the entire oil phase O while stirring, and emulsified by high-energy emulsification at a rotation speed of 15,000 rpm for 2 minutes, followed by ultrasonic homogenization at 450 W for 15 minutes to obtain colostrum W1 / O; the volume ratio of the oil phase O to the internal aqueous phase W1 is 5:5 or 6:4 or 7:3 or 8:2 or 9:1.

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

[0101] (5) Preparation of W1 / O / W2 double emulsion: Colostrum W1 / O and external water phase W2 were mixed at a volume ratio of 5:5, and emulsified by high-energy emulsification method at 10,000 rpm for 2 min to obtain double emulsion W1 / O / W2. The mixture was stored at room temperature to obtain W1 / O / W2 double emulsion containing brain polypeptide freeze-dried powder.

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

[0103] Table 4 Effect of oil-water ratio on the embedding efficiency of W1 / O / W2 emulsions for embedding brain polypeptide freeze-dried 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 double emulsion prepared with an oil-water ratio of 8:2 has the highest embedding rate.

[0106] Example 5 Effect of the Type of Hydrophilic Emulsifier on the Encapsulation Efficiency of W1 / O / W2 Double Emulsion Encapsulating Brain Polypeptide Lyophilized Powder

[0107] (1) Preparation of internal aqueous phase W1: After the brain polypeptide freeze-dried powder is mixed with ultrapure water, vortexed to make it evenly dispersed, and filtered through a 0.22 μm filter membrane to form an internal aqueous phase W1. The loading concentration of the brain polypeptide freeze-dried powder in the internal aqueous phase W1 is 25 mg / mL.

[0108] (2) Preparation of oil phase O: 6% PGPR was added to a certain amount of soybean oil and magnetically stirred at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, i.e., oil phase O.

[0109] (3) Preparation of colostrum W1 / O: The entire internal aqueous phase W1 was added dropwise into the entire oil phase O while stirring, and emulsified by high-energy emulsification at 15,000 rpm for 2 min, followed by ultrasonic homogenization at 450 W for 15 min to obtain colostrum W1 / O; the volume ratio of the oil phase O to the internal aqueous phase W1 was 8:2.

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

[0111] (5) Preparation of W1 / O / W2 double emulsion: Colostrum W1 / O and external water phase W2 were mixed at a volume ratio of 5:5, and emulsified by high-energy emulsification method at a rotation speed of 10,000 rpm for 2 minutes to obtain double emulsion W1 / O / W2. The mixture was stored at room temperature to obtain W1 / O / W2 double emulsion containing brain polypeptide freeze-dried powder.

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

[0113] Table 5 Effect of hydrophilic emulsifier type on the embedding efficiency of W1 / O / W2 emulsions for embedding brain polypeptide freeze-dried powder

[0114]

[0115] As shown in Table 5, the W1 / O / W2 double emulsion prepared with Tween 80 as the hydrophilic emulsifier has the highest embedding rate.

[0116] Example 6 Effect of Tween 80 Addition Amount on the Embedding Efficiency of W1 / O / W2 Double Emulsion Encapsulating Brain Polypeptide Lyophilized Powder

[0117] (1) Preparation of internal aqueous phase W1: After the brain polypeptide freeze-dried powder is mixed with ultrapure water, vortexed to make it evenly dispersed, and filtered through a 0.22 μm filter membrane to form an internal aqueous phase W1. The loading concentration of the brain polypeptide freeze-dried powder in the internal aqueous phase W1 is 25 mg / mL.

[0118] (2) Preparation of oil phase O: 6% PGPR was added to a certain amount of soybean oil and magnetically stirred at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, i.e., oil phase O.

[0119] (3) Preparation of colostrum W1 / O: The entire internal water phase W1 is added dropwise into the entire oil phase O while stirring, and emulsified by high-energy emulsification at a rotation speed of 15,000 rpm for 2 minutes, followed by ultrasonic homogenization at 450 W for 15 minutes to obtain colostrum W1 / O; the volume ratio of the oil phase O to the internal water phase W1 is 8:2.

[0120] (4) Preparation of external aqueous phase W2: Preheat a certain volume of ultrapure water in a 50°C water bath for 15 min, add different proportions of Tween 80 (1%, 2%, 4%, 6%, 8%, or 10%) and 0.1% sucrose fatty acid ester, and stir at 360 rpm for 30 min to fully dissolve them, which is the external aqueous phase W2.

[0121] (5) Preparation of W1 / O / W2 double emulsion: Colostrum W1 / O and external water phase W2 were mixed at a volume ratio of 5:5, and emulsified by high-energy emulsification method at a rotation speed of 10,000 rpm for 2 minutes to obtain double emulsion W1 / O / W2. The mixture was stored at room temperature to obtain W1 / O / W2 double emulsion containing brain polypeptide freeze-dried powder.

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

[0123] Table 6 Effect of Tween 80 addition on the embedding efficiency of W1 / O / W2 emulsions for embedding brain polypeptide freeze-dried powder

[0124] Tween80 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 double emulsion prepared with 4% Tween 80 as the hydrophilic emulsifier has the highest embedding rate.

[0126] Example 7 Effect of the internal-external ratio on the embedding efficiency of W1 / O / W2 double emulsion for embedding brain polypeptide freeze-dried powder

[0127] (1) Preparation of internal aqueous phase W1: After the brain polypeptide freeze-dried powder is mixed with ultrapure water, vortexed to make it evenly dispersed, and filtered through a 0.22 μm filter membrane to form an internal aqueous phase W1. The loading concentration of the brain polypeptide freeze-dried powder in the internal aqueous phase W1 is 25 mg / mL.

[0128] (2) Preparation of oil phase O: 6% PGPR was added to a certain amount of soybean oil and magnetically stirred at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, i.e., oil phase O.

[0129] (3) Preparation of colostrum W1 / O: The entire internal aqueous phase W1 was added dropwise into the entire oil phase O while stirring, and emulsified by high-energy emulsification at 15,000 rpm for 2 min, followed by ultrasonic homogenization at 450 W for 15 min to obtain colostrum W1 / O; the volume ratio of the oil phase O to the internal aqueous phase W1 was 8:2.

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

[0131] (5) Preparation of W1 / O / W2 double emulsion: Colostrum W1 / O and external water phase W2 were mixed at a certain volume ratio (5:5 or 4:6 or 3:7 or 2:8 or 1:9), and emulsified by high-energy emulsification method at 10,000 rpm for 2 min to obtain double emulsion W1 / O / W2. The mixture was stored at room temperature to obtain W1 / O / W2 double emulsion containing brain polypeptide freeze-dried powder.

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

[0133] Table 7 Effect of internal and external ratio on the embedding efficiency of W1 / O / W2 emulsions for embedding brain polypeptide freeze-dried powder

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

[0135] It can be seen from Table 7 that the W1 / O / W2 double emulsion prepared with an internal-external ratio of 4:6 has the highest embedding rate.

[0136] Example 8 Preparation of High Stability W1 / O / W2 Double Emulsion

[0137] (1) Preparation of internal aqueous phase W1: After the brain polypeptide freeze-dried powder is mixed with ultrapure water, vortexed to make it evenly dispersed, and filtered through a 0.22 μm filter membrane to form an internal aqueous phase W1. The loading concentration of the brain polypeptide freeze-dried powder in the internal aqueous phase W1 is 25 mg / mL.

[0138] (2) Preparation of oil phase O: 6% PGPR was added to a certain amount of soybean oil and magnetically stirred at 320 rpm for 10 min to obtain 6% lipophilic emulsifier, i.e., oil phase O.

[0139] (3) Preparation of colostrum W1 / O: The entire internal aqueous phase W1 was added dropwise into the entire oil phase O while stirring, and emulsified by high-energy emulsification at 15,000 rpm for 2 min, followed by ultrasonic homogenization at 450 W for 15 min to obtain colostrum W1 / O; the volume ratio of the oil phase O to the internal aqueous phase W1 was 8:2.

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

[0141] (5) Preparation of high-stability W1 / O / W2 double emulsion: Colostrum W1 / O and external aqueous phase W2 were mixed at a volume ratio of 4:6, and emulsified by high-energy emulsification method at 10,000 rpm for 2 min to obtain double emulsion W1 / O / W2. After storage at room temperature, a high-stability W1 / O / W2 double emulsion containing brain polypeptide lyophilized powder was obtained.

[0142] Example 9 Evaluation of High Stability W1 / O / W2 Double Emulsion Encapsulating Lyophilized Brain Polypeptide Powder

[0143] 1. Observation of emulsion appearance

[0144] The W1 / O / W2 double emulsions with different xanthan gum addition amounts prepared in Example 8 were added into 20 mL vials, sealed, capped, and left to stand at room temperature for 30 days to observe the changes in the emulsion appearance. Figure 2 As shown: the freshly prepared emulsion is uniform and fine, without impurities, flocculation, and stratification. After standing at room temperature for 30 days, the emulsion without xanthan gum showed obvious stratification, the emulsion with 0.2% xanthan gum had a slight stratification at the bottom, and the other emulsions showed no obvious changes.

[0145] 2. Droplet morphology observation

[0146] Use an optical microscope to observe the highly stable W1 / O / W2 double emulsion. Pipette 2.5μL of each sample onto a glass slide, dilute 5 times with ultrapure water directly on the microscope slide, and then gently cover the emulsion with a cover glass. Use a 40x objective lens to observe the W1 / O / W2 emulsion. The results are as follows: Figure 3 As shown, the particle size of the emulsion without xanthan gum is large, which is not conducive to the stability of the emulsion, and the particle size of the emulsion with xanthan gum is reduced. Among them, the emulsion with 0.8% xanthan gum has a small particle size and more double-pack structures, which is the best choice.

[0147] 3. Determination of embedding rate of brain polypeptide freeze-dried powder

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

[0149]

[0150] C——Concentration of brain polypeptide freeze-dried powder that migrated to the external aqueous phase;

[0151] V——the volume of the supernatant;

[0152] C0——the concentration of brain polypeptide freeze-dried powder added to the double emulsion;

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

[0154] V0 – volume of double emulsion.

[0155] The results are as follows Figure 4 As shown, from Figure 4 It can be seen that the emulsion embedding rate of different xanthan gum addition amounts is high, indicating that xanthan gum helps to stabilize the W1 / O / W2 double emulsion system, among which the embedding rate is the highest when 0.8% xanthan gum is added.

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

[0157] 20 μL of each of the emulsions prepared in Example 8 with different amounts of xanthan gum added was taken and added to 4 mL of 0.1% sodium dodecyl sulfate (SDS) solution and mixed evenly. The absorbance A0 at a wavelength of 500 nm was measured. After standing for 60 min, the absorbance A 60 The emulsion activity index (EAI) and emulsion stability index (ESI) are calculated using the following formulas.

[0158]

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

[0160] N is the dilution multiple;

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

[0162] θ is the volume of oil phase;

[0163] ΔT is the time difference, min;

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

[0165] The results are as follows Figure 5 As shown in the figure, the EAI and ESI of the emulsion with xanthan gum added are significantly higher than those without xanthan gum. The emulsification property of protein depends on its ability to reduce the interfacial tension between the two phases. With the increase of xanthan gum addition in the system, the change trends of EAI and ESI are roughly similar. When the xanthan gum addition is 0.8%, EAI and ESI are both the largest, reaching 15.282m 2 / g and 452.2min, which shows that the interfacial properties and stability are better than those of other xanthan gum addition amounts.

[0166] 5. Storage stability determination

[0167] Double emulsions may be stratified during storage, with the upper layer being the emulsion retention layer and the lower layer being the transparent or turbid layer. The newly prepared double emulsions were placed in vials and stored at room temperature for 0 days, 0.5 days, 1 day, 3 days, 7 days, 14 days, and 30 days, and the emulsion stratification was observed. Figure 6 The stability index calculation formula is:

[0168]

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

[0170] H0——latex sample height, cm

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

[0172] After standing at room temperature for 30 days, it was found that the emulsion without xanthan gum was stratified within 12 hours. From day 0 to day 7, the double emulsions with 0.2% to 1.0% xanthan gum were in good condition without stratification. On day 30, it was found that except for the double emulsion with 0.2% xanthan gum, which was slightly stratified, the other emulsions were in good condition without stratification.

[0173] 6. Centrifugal Retention Rate Determination

[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 is to use centrifugal means to separate the prepared double emulsions under the action of centrifugal force. The calculated retention rate of the emulsion layer is used as a measure of the centrifugal stability of the double emulsion. Take 1mL of the freshly prepared double emulsion and put it into a 1.5mL centrifuge tube. Centrifuge it at 2000rpm for 10min. The double emulsion will separate into layers and the outer water phase will precipitate at the bottom. Measure the total volume of the emulsion in the centrifuge tube and the volume of the precipitated water phase. The results are as follows: Figure 7 The centrifugal retention rate of W1 / O / W2 double emulsion was calculated according to the formula.

[0175]

[0176] Where V t Represents the total volume of the liquid layer in the centrifuge tube, V c It indicates the volume of the aqueous phase separated after separation.

[0177] The centrifugal retention rate of the emulsion without xanthan gum was only 50.2%, indicating that the emulsion system without xanthan gum was unstable and prone to stratification, which was consistent with the results of the static observation of the surface diagram and storage stability. The centrifugal retention rate of the emulsion with 0.2% xanthan gum added was 53.5%, which was not much different from the emulsion without xanthan gum added. The centrifugal retention rate of the emulsion with other xanthan gum additions was 100%, indicating good stability.

[0178] Example 10 In vitro evaluation of W1 / O / W2 double emulsions encapsulating lyophilized brain polypeptide powder

[0179] In vitro stability of brain peptide freeze-dried powder under simulated gastrointestinal fluid conditions.

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

[0181] (2) Simulated intestinal fluid (SIF) digestion: The simulated intestinal fluid contained 1.5 mL of saline 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 enzymes and bile salts were dissolved in phosphate buffered saline (pH 7.0). 1 mL of SIF was added to the pH-adjusted chyme, and then the pH value of the system was monitored and maintained at 7.0 by 0.1 M NaOH solution. The digestion was continued for 2 h at 37 °C with a magnetic stirring speed of 100 rpm. The volume of NaOH consumed was recorded. The percentage of free fatty acid (FFA) release was calculated by the following formula:

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

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

[0184] After passing through simulated gastric juice, the unencapsulated brain polypeptide freeze-dried powder lost its activity due to pepsin and low pH conditions, while the brain polypeptide freeze-dried powder encapsulated by the double emulsion retained its activity. After the emulsion enters the small intestine digestion stage, under the action of lipase, the oil phase will decompose into glycerol and FFA. The FFA release rate of the W1 / O / W2 emulsion increases first and then tends to be flat as the digestion process progresses. The results are shown in Figure 8 . After 120 minutes of digestion, the FFA release rate of the unencapsulated brain polypeptide freeze-dried powder in the hydrolyzate of simulated 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 double emulsion with xanthan gum can reach 80% to 90%, among which the FFA release rate of the W1 / O / W2 emulsion with 0.8% xanthan gum added is the highest, which is 91.5%, indicating that the brain polypeptide freeze-dried powder encapsulated in the double emulsion can stably reach the intestine after passing through the gastric juice and is efficiently released in the intestine. The release rate is 45.17% higher than that of the W1 / O / W2 emulsion without xanthan gum.

[0185] (3) Chemical stability (S * ) and bioaccessibility (B * ): The final chyme remaining after small intestinal digestion was centrifuged at 10,000 × g for 30 min and separated into three layers: bottom sediment phase, middle micelle phase and upper oil phase. * ) and bioaccessibility (B * ) is measured according to the following formula. The results are as follows Fig. 9 shown.

[0186] %S*=C Digesta / C Initial ×100

[0187] %B*=C Micelle / C Initial ×100

[0188] Among them C Initial is the initial concentration of brain polypeptide freeze-dried powder in the emulsion, C Digesta is the concentration of brain peptide freeze-dried powder in the total digesta collected after the small intestine stage, C Micelle It is the concentration of brain polypeptide freeze-dried powder in the mixed micelle phase.

[0189] The results of the study on simulated gastrointestinal digestion showed that the chemical stability and bioaccessibility of W1 / O / W2 double emulsions with different amounts of xanthan gum added were improved compared with the unencapsulated hydrolyzate. Among them, the chemical stability of the sample with 0.8% xanthan gum was improved by 39.03% and the bioaccessibility was improved by 86.60% compared with the hydrolyzate. The results showed that W1 / O / W2 double emulsions encapsulating brain peptide freeze-dried powder can prevent them from being released in the stomach. The reason for reducing bitterness and improving gastrointestinal digestion stability is that the peptides are dispersed in the aqueous phase, and a layer of fat is wrapped around the water droplets. W1 / O / W2 double emulsions have the effect of masking bitterness and improving gastrointestinal stability, which can expand the application of bioactive ingredients in food.

[0190] In summary, 0.8% xanthan gum was selected to prepare a high-stability W1 / O / W2 double emulsion.

[0191] Example 11 Preparation of Fruity Brain Polypeptide Oral Liquid

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

[0193] (2) adding 4% concentrated peach juice and 0.1% concentrated lemon juice, stirring evenly, and adjusting the pH to 7.0 to obtain a prepared mixed solution;

[0194] (3) filtering the prepared mixed solution through a 500-mesh filter cloth, and then filling it to obtain an oral solution;

[0195] (4) sterilizing the oral liquid filled in step (3) at high temperature and high pressure, wherein the sterilization method is: sterilizing with high-pressure steam at 121° C. for 15 min, and cooling until the core temperature is less than 35° C. to obtain the finished product.

[0196] (5) The sensory evaluation scoring method was used to conduct sensory evaluation on the brain polypeptide oral liquid. Four indicators, namely smell and taste, color, tissue morphology and impurities, were selected, and 10 trained students were asked to conduct sensory evaluation on the obtained fruity brain polypeptide oral liquid. The evaluation criteria are shown in Table 2, and the evaluation results are shown in Table 3.

[0197] The difference between Example 12 and Example 11 is that in step (2), 3% mango concentrated juice and 0.5% orange concentrated clear juice are used.

[0198] The difference between Example 13 and Example 11 is only that in step (2), the apple concentrated juice is 4% and the peach concentrated juice is 1%.

[0199] The difference between Example 14 and Example 11 is only that the grape concentrated juice in step (2) is 3%.

[0200] Table 2 Flavor evaluation criteria for brain polypeptide oral solution

[0201]

[0202] Table 3 Sensory evaluation results of brain polypeptide oral liquid

[0203]

[0204] It can be seen from the sensory evaluation results that the brain polypeptide freeze-dried powder aqueous solution has a bitter taste due to the exposure of hydrophobic amino acids in pig brain protein powder during hydrolysis, which causes it to directly bind to bitter receptors in the mouth, resulting in a significant bad smell and difficult to swallow. In addition, due to the small amount of microparticles in the brain polypeptide freeze-dried powder aqueous solution that permeates the filter membrane, they are foamed and expanded in the water, turning into floating objects visible to the naked eye, which leads to a foreign body sensation, and the comprehensive score is the lowest, only 59 points. The W1 / O / W2 type double emulsion (Example 8) effectively masks this bad smell, but due to the use of emulsifiers, it brings a slight special smell. In order to reduce the influence of the bad flavor of the emulsifier and enrich the flavor of the brain polypeptide oral liquid, the present invention optimizes its flavor by adding concentrated fruit juice. It can be seen from the sensory evaluation results that various fruit-flavored brain polypeptide oral liquids (Examples 11 to 14) have a good taste, strong palatability, sweet and sour without being greasy, and obviously cover up the bad flavor, improve the sensory quality of the beverage, and improve consumer acceptance.

Claims

1. A method for preparing a high-stability W1 / O / W2 double emulsion for embedding freeze-dried brain polypeptide powder, characterized in that: The following steps are involved: S1: Preparation of inner aqueous phase W1: After mixing the brain polypeptide lyophilized powder with ultrapure water, vortex to make it evenly dispersed, and filter through a 0.22 μm filter membrane to form an inner aqueous phase W1, wherein the loading amount of the brain polypeptide lyophilized powder is 25 mg / mL; S2: Preparation of oil phase O: dissolving the lipophilic emulsifier in edible oil and forming oil phase O through magnetic stirring; S3: Preparation of colostrum W1 / O: Preliminarily mix the oil phase O and the internal water phase W1, and then stir, homogenize with a homogenizer and homogenize with ultrasound to obtain water-in-oil colostrum W1 / O; S4: Preparation of external aqueous phase W2: Add hydrophilic emulsifier and sucrose fatty acid ester into ultrapure water, stir and dissolve at 50°C for 30 minutes to obtain external aqueous phase W2; S5: Preparation of W1 / O / W2 type double emulsion: adding the colostrum W1 / O described in S3 dropwise into the external aqueous phase W2, and obtaining a W1 / O / W2 type double emulsion encapsulating the freeze-dried brain polypeptide powder through stirring, high-speed shearing, homogenization and ultrasonic emulsification; S6: Preparation of high-stability W1 / O / W2 double emulsion: Add a stabilizer to the W1 / O / W2 double emulsion described in S5, and after stirring, homogenizing, and ultrasonic emulsification, a high-stability W1 / O / W2 double emulsion can be obtained.

2. The method according to claim 1, characterized in that In step S2, the edible oil is soybean oil, the lipophilic emulsifier is polyglycerol ricinoleate (PGPR), and the amount of the lipophilic emulsifier added is 1-10%.

3. The method according to claim 1, characterized in that The mass ratio of the oil phase O to the internal water phase W1 in step S3 is one of 5:5, 6:4, 7:3, 8:2, and 9:1; the homogenization condition in step S3 is shearing at 10000-15000 rpm for 2 min, and the ultrasonic condition is 450 W ultrasonication for 15 min.

4. The method according to claim 1, characterized in that The preparation of W2 in step S4 is to add a hydrophilic emulsifier and sucrose fatty acid ester into ultrapure water and stir to fully dissolve, wherein 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; the amount of the hydrophilic emulsifier added to W2 in step S4 is 1-10%, and the concentration of the sucrose fatty acid ester is 0.1%.

5. The method according to claim 1, characterized in that In step S5, the volume ratio of the external water phase W2 to the colostrum W1 / O is one of 5:5, 6:4, 7:3, 8:2, and 9:

1.

6. The method according to claim 1, characterized in that The stabilizer in step S6 is xanthan gum, and the concentration of the xanthan gum is 0.2-1%.

7. A high-stability W1 / O / W2 double emulsion for embedding freeze-dried brain polypeptide powder, characterized in that: The compound is prepared by the preparation method according to any one of claims 1 to 6.

8. Use of the emulsion according to any one of claims 1 to 6 in the preparation of fruity brain polypeptide oral liquid.

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