An animal fat tissue analog of lipid-chitosan-based pickering high internal phase emulsion and a preparation method and application thereof

Abstract

The application discloses an animal fat tissue simulator of lipid-chitosan-based Pickering high internal phase emulsion and a preparation method thereof. The preparation method is as follows: (1) chitosan and an emulsifier are dissolved in a solvent by heating to obtain an aqueous phase, food-grade lipids are melted by heating to obtain an oil phase, the aqueous phase is slowly poured into the oil phase and stirred, and then the mixture is subjected to high-speed shearing, ultrasonic treatment and cooling to obtain a chitosan-coated lipid nanoparticle dispersion; (2) the above particle dispersion is mixed with edible vegetable oil, and then subjected to high-speed shearing to obtain the lipid-chitosan-based Pickering high internal phase emulsion. The application has the advantages of convenient operation, simple conditions, and the like. The lipid-chitosan-based Pickering high internal phase emulsion as the animal fat tissue simulator has the characteristics of green health, compact structure, heat melting, close mouth feeling to real animal fat, and the like, and can be widely applied to the food and pharmaceutical industries.

Description

Technical Field

[0001] The invention belongs to the technical field of emulsion processing, and particularly relates to an animal fat tissue mimic of a lipid-chitosan-based Pickering high internal phase emulsion, and a preparation method and application thereof. Background Art

[0002] As an integral component of meat products, animal adipose tissue provides excellent texture, flavor, mouthfeel, and textural properties. It consists of connective tissue composed of collagen and embedded adipocytes containing crystals and liquid triglycerides. When exposed to the heat of cooking, adipose tissue partially melts and flows due to the destruction of collagen and the weakening of the crystal network, making it a prominent contribution to the sensory experience of food, such as juiciness and tenderness. With the growth of the market for green and sustainable biomimetic meat products, the development of animal fat mimics has become increasingly important. Structured emulsions have become an effective strategy for preparing fat mimics, but the general lack of melting properties is one of the issues that limit their ability to simulate animal adipose tissue.

[0003] Pickering high internal phase emulsion is a new type of structured emulsion that uses solid particles to stabilize the oil-water system and has an internal phase volume accounting for more than 74%. The water-in-oil type Pickering high internal phase emulsion is structurally endowed with a good semi-solid plastic state by the network of aggregated oil droplets, and has great application prospects in the market for healthy animal fat tissue simulants. Lipid nanoparticles are formed by heating and melting food-grade lipids, including beeswax, and then cooling and crystallizing. They are usually highly hydrophobic and require surface modification to adsorb and assemble at the oil-water interface, hindering the aggregation and precipitation of oil droplets in the emulsion. However, when the temperature rises, the remelting of the lipid base can also cause reverse instability of the interface. Patent CN202211031308.3 confirms that hydrophilic proteins are more conducive to the stable construction of Pickering high internal phase emulsions with lipid nanoparticles than hydrophilic small molecules. However, they usually have the characteristics of high-temperature denaturation and aggregation. After heating, they stretch and cross-link in the external phase, hindering the precipitation of liquid oil. Therefore, this Pickering high internal phase emulsion cannot further imitate the melting behavior of animal fat tissue. Polysaccharides are mainly hydrophilic macromolecules, among which cationic chitosan is not easily denatured by heat and can also undergo electrostatic assembly with negatively charged lipid nanoparticles, which can effectively improve the dispersibility of the particles. The use of polysaccharide-coated lipid nanoparticles to prepare Pickering high internal phase emulsions is a novel development idea for animal fat tissue mimics, and related research has not yet been reported. However, there are large differences in the molecular structures of polysaccharides, and it is impossible to directly determine whether chitosan can enable Pickering high internal phase emulsions to achieve the oil separation performance and lubricating taste of real animal fat tissue. In addition, under different emulsifier concentration conditions, the crystal structure and interfacial properties of chitosan-coated lipid nanoparticles will change accordingly, and the influence of this on the structure and properties of Pickering high internal phase emulsion fat mimics also needs to be further determined. Summary of the Invention

[0004] In order to overcome the deficiencies and shortcomings of the prior art, the primary purpose of the present application is to provide a lipid-chitosan-based Pickering high internal phase emulsion animal fat tissue simulator, which has the characteristics of green health, tight structure, heat melting and close mouthfeel to real animal fat.

[0005] Another purpose of the present application is to provide a preparation method of the animal fat tissue simulator.

[0006] Another purpose of the present application is to provide the application of the animal fat tissue simulator in the field of food and / or medicine.

[0007] The above purposes of the present application are achieved by the following technical solutions:

[0008] The present application provides a lipid-chitosan-based Pickering high internal phase emulsion animal fat tissue simulator, which uses a lipid-chitosan-based Pickering high internal phase emulsion as an animal fat tissue simulator, mainly comprising:

[0009] (1) a particle dispersion liquid;

[0010] (2) an edible vegetable oil;

[0011] The above animal fat tissue simulator of the present application is prepared by homogenously compounding the particle dispersion liquid and the edible vegetable oil, and has partial structural similarity to real animal fat tissue. After the oil droplets are separated by a thin layer of water phase and tightly packed into polyhedral shape, they become "fat cells" surrounded by a particle interface film. The particle interface film has good mechanical strength and temperature sensitivity, so that the animal fat tissue simulator can more closely approach the mechanical and thermal properties of animal fat tissue under low saturated fatty acid and low surfactant conditions, which can not only meet people's pursuit of nutritional health, but also bring people a unique eating experience.

[0012] The present application also provides a preparation method of the above animal fat tissue simulator, comprising the following steps:

[0013] (1) dissolving chitosan and an emulsifier in a solvent by heating to obtain an aqueous phase, and then melting food-grade lipids to obtain an oil phase, and then adding the aqueous phase into the oil phase and stirring, followed by high-speed shearing, ultrasonic treatment and cooling treatment to obtain a chitosan-coated lipid nanoparticle dispersion liquid;

[0014] (2) adding an edible vegetable oil to the chitosan-coated lipid nanoparticle dispersion liquid, and then performing high-speed shearing and rapid cooling to obtain the lipid-chitosan-based Pickering high internal phase emulsion animal fat tissue simulator;

[0015] The concentration of the chitosan in the lipid-chitosan-based Pickering high-internal-phase emulsion in step (1) is 0.5-0.9 g / 100 mL; the concentration of the emulsifier in the lipid-chitosan-based Pickering high-internal-phase emulsion is 1-2 g / 100 mL; and the concentration of the food-grade lipid in the lipid-chitosan-based Pickering high-internal-phase emulsion is 1-3 g / 100 mL.

[0016] The food-grade lipid in step (1) is a lipid with a melting point ≥ 42 ℃.

[0017] The volume ratio of the particle dispersion liquid to the edible vegetable oil in step (2) is 20-30:70-80.

[0018] Preferably, the emulsifier in step (1) is Tween; the solvent is a 1-3 g / 100 mL ice acetic acid solution; the food-grade lipid is at least one of beeswax, palm stearin, and fully hydrogenated vegetable oil; and the edible vegetable oil is at least one of fish oil, corn oil, rapeseed oil, soybean oil, peanut oil, camellia oil, safflower oil, olive oil, walnut oil, sunflower seed oil, and flaxseed oil.

[0019] Preferably, the food-grade lipid is beeswax.

[0020] Preferably, the heating in step (1) refers to heating the water phase and the oil phase to a similar temperature under the premise of ensuring that the food-grade lipid is completely melted; the heating conditions for the water phase are a temperature of 70-85 ℃ and a time of 60-90 min; and the heating conditions for the oil phase are a temperature of 70-85 ℃ and a time of 20-35 min; the stirring in step (1) is magnetic stirring; and the stirring conditions are a rotation speed of 100-200 rpm and a time of 3-8 min.

[0021] Preferably, the high-speed shearing in step (1) is performed at a rotation speed of 10,000-20,000 rpm for 1-5 min.

[0022] Preferably, the ultrasonic treatment in step (1) is performed at a power of 100-500 W for 5-10 min.

[0023] Preferably, the cooling in step (1) is performed at a temperature of 4-37 ℃ for ≥ 24 h.

[0024] Preferably, the high-speed shearing in step (2) is performed at a rotation speed of 10,000-20,000 rpm for 2-5 min.

[0025] Compared with the prior art, the present application has the following advantages and beneficial effects:

[0026] (1) The animal fat tissue mimic of the lipid-chitosan-based Pickering high internal phase emulsion of the present invention uses food-grade lipid as the particle base material, has convenient and mild preparation conditions, is easy to achieve large-scale production, does not involve toxic and harmful reagents, and expands the application prospects of the Pickering particle-stabilized high internal phase emulsion system in the fields of food, medicine and cosmetics.

[0027] (2) The animal fat tissue mimic of the lipid-chitosan-based Pickering high internal phase emulsion of the present invention uses chitosan-coated lipid nanoparticles that can be controllably self-assembled onto the oil / water surface to form a monolayer or multilayer fat crystal interface film to stably disperse droplets, and the interface structure adsorbed by the lipid Pickering particles will be conducive to the construction of an oil-in-water high internal phase emulsion.

[0028] (3) Compared with the commonly used protein-based Pickering emulsion, the animal fat tissue mimetic of the lipid-chitosan-based Pickering high internal phase emulsion of the present invention has better temperature sensitivity and can be used to simulate the hot melting behavior required by animal fat tissue. It has great development value in the market of healthy food fat mimetic. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Inverted macroscopic images and optical microscopic images of animal adipose tissue mimics of the food-grade Pickering high internal phase emulsions in Examples 1 to 4 and Comparative Examples 1 to 4;

[0030] Figure 2 These are graphs showing the appearance changes of the animal fat tissue simulant of the food-grade Pickering high internal phase emulsion and commercially available pig back fat tissue before and after heating in Examples 1 to 4 and Comparative Example 1;

[0031] Figure 3 The rheological temperature scanning curve of the animal fat tissue simulant of the food-grade Pickering high internal phase emulsion in Example 1 and Comparative Example 1 before cooking;

[0032] Figure 4 The oral soft tribology curves of the animal fat tissue simulant of the food-grade Pickering high internal phase emulsion in Examples 1 to 4 and Comparative Example 1 and commercially available pig back fat tissue after cooking are shown. DETAILED DESCRIPTION

[0033] The present invention will be further described in detail below with reference to specific examples, but the embodiments of the present invention are not limited thereto. The materials used in the examples of the present invention can all be purchased commercially.

[0034] In the following examples, 1 part by mass refers to 1 g. The unit of solution concentration (w / v) is g / mL.

[0035] Example 1

[0036] Preparation of chitosan-coated lipid nanoparticles: 3.5 parts by mass of chitosan and 4 parts by mass of Tween 40 were added to 100 mL of a 2% w / v ice acetic acid solution to obtain an aqueous phase by placing it in a water bath at 80°C for 60-90 min until complete dissolution. 10 parts by mass of beeswax was placed in a water bath at 80°C for 20-35 min until complete melting to obtain an oil phase. Then, the aqueous phase was slowly poured into the oil phase at a similar temperature, stirred at 150 rpm for 5 min, sheared at a speed of 14000 rpm for 1 min using a high-speed shearing machine, and then ultrasonically treated at a power of 450 W for 7 min. It was placed in a 4°C water bath and stirred to cool for 1 h, then taken out and stored in a 4°C refrigerator for 24 h for standby, and chitosan-coated lipid nanoparticles were prepared.

[0037] Preparation of lipid-chitosan-based Pickering high internal phase emulsion: The above particle dispersion liquid was mixed with edible soybean oil at a volume ratio of 25:75, first treated with a high-speed shearing machine at 14000 rpm for 2 min, then quickly cooled and stored in a 4°C refrigerator for 24 h for standby, and an oil-in-water type lipid-chitosan-based Pickering high internal phase emulsion was prepared.

[0038] Optical microscopic observation of the prepared lipid-chitosan-based Pickering high internal phase emulsion: Freshly prepared samples were spotted on a glass slide using a capillary tube, and the morphology was observed using an optical microscope.

[0039] Thermostatic cooking test of the prepared lipid-chitosan-based Pickering high internal phase emulsion: Freshly prepared samples were placed in a convection oven at 70°C and heated for 15 min, and then the melting behavior was recorded.

[0040] Rheological temperature scanning of the prepared lipid-chitosan-based Pickering high internal phase emulsion: A MCR 702e MultiDrive rheometer and a PP25 flat clamp were used to determine the thermal responsiveness of the emulsion, and the temperature range was 4-80°C.

[0041] Oral soft tribological evaluation of the prepared lipid-chitosan-based Pickering high internal phase emulsion cooking: A MCR 702e MultiDrive rheometer and a T-PTD200 ball three-pin accessory were used to simulate oral soft friction to determine the lubrication properties of the cooking at 37°C, and the simulated oral stress was 1.00 N and the sliding speed was 0.1-10 mm / s.

[0042] The food-grade lipid-chitosan-based Pickering high internal phase emulsion of this example was stored in a 4°C refrigerator for 24 h, and the whole system was semi-solid, which could be inverted without flowing in the container. For details, please refer to Figure 1 . The high internal phase emulsion of this example was observed by optical microscope, and the droplet distribution was relatively uniform. The strong interaction between particles caused the emulsion to form a 3D network gel structure, which helped to delay the occurrence of creaming phenomenon Figure 1 . FromFigure 2 As can be seen, the commercially available pork back fat tissue has a significant collapse and oil bleeding behavior after cooking, while the high internal phase emulsion of this example has a similar appearance with the pork back fat at room temperature, and can also transform from a semi-solid to a flowable state after heating, with some soybean oil bleeding out around the cooked sample, due to the fact that the granular matrix, beeswax, will melt at temperatures above 60°C, and the emulsion interface structure will be destroyed, losing the ability to stabilize the internal phase liquid oil. Figure 3 ) However, this oil bleeding behavior gives the cooked sample a good mouth lubrication sensation. Figure 4

[0043] Example 2

[0044] Preparation of chitosan-coated lipid nanoparticles: 3.5 parts by mass of chitosan and 4 parts by mass of Tween 40 were added to a 100 mL 2% w / v glacial acetic acid solution, which was placed in a 80°C water bath for 60-90 min to completely dissolve to obtain an aqueous phase, 5 parts by mass of beeswax was placed in a 80°C water bath for 20-35 min to completely melt to obtain an oil phase, then the aqueous phase was slowly poured into the oil phase at a similar temperature, stirred at 150 rpm for 5 min, sheared at a speed of 14000 rpm for 1 min with a high-speed shearing machine, and then treated with ultrasound at a power of 450 W for 7 min, placed in a 4°C water bath and stirred to cool for 1 h, then taken out and stored in a 4°C refrigerator for 24 h for standby, to prepare chitosan-coated lipid nanoparticles.

[0045] Preparation of lipid-chitosan-based Pickering high internal phase emulsion: the above particle dispersion liquid was mixed with edible soybean oil at a volume ratio of 25:75, first treated with a high-speed shearing machine at 14000 rpm for 2 min, then quickly cooled and stored in a 4°C refrigerator for 24 h for standby, to prepare an oil-in-water type lipid-chitosan-based Pickering high internal phase emulsion.

[0046] Optical microscopic observation of the prepared lipid-chitosan-based Pickering high internal phase emulsion: the freshly prepared sample was spotted on a glass slide using a capillary tube, and the morphology was observed using an optical microscope.

[0047] Thermostatic cooking test of the prepared lipid-chitosan-based Pickering high internal phase emulsion: the freshly prepared sample was placed in a convection oven at 70°C and heated for 15 min, and then the melting behavior was recorded.

[0048] Oral soft tribological evaluation of the cooked sample of the prepared lipid-chitosan-based Pickering high internal phase emulsion: the MCR702e MultiDrive rheometer and T-PTD200 ball three-pin accessory were used to simulate oral soft friction to determine the lubrication properties of the cooked sample at 37°C, with a simulated oral stress of 1.00 N and a sliding speed of 0.1-10 mm / s.

[0049] ​The food-grade lipid-chitosan-based Pickering high internal phase emulsion of the present example was stored in a 4℃ refrigerator for 24h, similar to Example 1, and the whole system was semi-solid and could not flow when the container was inverted. Figure 1 The high internal phase emulsion of the present example was observed by optical microscopy, and the droplet distribution was not as uniform as that of Example 1. The decrease in particle concentration would weaken the density of the interfacial barrier film, resulting in a small amount of oil droplet coalescence during the collision process during storage. Figure 1 Figure 2 The high internal phase emulsion of the present example had temperature responsiveness, and could be converted from semi-solid to flowable state after heating. There was a small amount of soybean oil around the cooked product, similar to the melting behavior of commercially available pork back fat. However, the mouth lubrication of the cooked product of the present example was poorer than that of Example 1, and it could be seen that the decrease in lipid nanoparticle concentration might directly affect the eating experience. Figure 4

[0050] Example 3

[0051] Preparation of chitosan-coated lipid nanoparticles: 3 parts by mass of chitosan and 4 parts by mass of Tween 40 were added to 100 mL of a 2% w / v ice acetic acid solution, which was placed in a 80℃ water bath for 60-90 min to completely dissolve to obtain an aqueous phase. 10 parts by mass of beeswax was placed in a 80℃ water bath for 20-35 min to completely melt to obtain an oil phase. Then the aqueous phase was slowly poured into the oil phase at a similar temperature, stirred at 150 rpm for 5 min, sheared by a high-speed shearing machine at a speed of 14000 rpm for 1 min, and then ultrasonically treated at a power of 450 W for 7 min. It was placed in a 4℃ water bath and stirred to cool for 1 h, then taken out and stored in a 4℃ refrigerator for 24 h for standby, and chitosan-coated lipid nanoparticles were prepared.

[0052] Preparation of lipid-chitosan-based Pickering high internal phase emulsion: The above particle dispersion liquid was mixed with edible soybean oil at a volume ratio of 25:75, first treated by a high-speed shearing machine at 14000 rpm for 2 min, then quickly cooled and stored in a 4℃ refrigerator for 24 h for standby, and an oil-in-water type lipid-chitosan-based Pickering high internal phase emulsion was prepared.

[0053] Optical microscopic observation of the prepared lipid-chitosan-based Pickering high internal phase emulsion: The freshly prepared sample was spotted on a glass slide using a capillary tube, and the morphology was observed using an optical microscope.

[0054] Thermostatic cooking test of the prepared lipid-chitosan-based Pickering high internal phase emulsion: The freshly prepared sample was placed in a convection oven and heated at 70℃ for 15 min, and the melting behavior was recorded.

[0055] ​​Oral soft tribological evaluation of lipid-chitosan-based Pickering high internal phase emulsion cooking materials was performed: an MCR702e MultiDrive rheometer and a T-PTD200 ball tripod fitting were used to simulate oral soft friction to determine the lubrication properties of the cooking materials at 37°C. The simulated oral stress was 1.00 N and the sliding speed was 0.1-10 mm / s.

[0056] The food-grade lipid-chitosan-based Pickering high internal phase emulsion of this embodiment was placed in a refrigerator at 4°C for 24 hours. Similar to Examples 1 and 2, the entire system was in a semi-solid state and could be inverted in the container without flowing ( Figure 1 The high internal phase emulsion of this embodiment was observed under an optical microscope. The oil droplets were larger than those of Example 1 and Example 2 ( Figure 1 ), as the concentration of chitosan decreases, the positive charge on the particle surface decreases, causing it to aggregate, occupying a large interfacial area and resulting in low interfacial adsorption. Figure 2 The thermal behavior of commercially available pork back fat is similar to that of pork back fat in China. When heated, the semi-solid shape easily melts and cannot be maintained, and more liquid soybean oil is present around the cooked food. In this embodiment, the uneven large oil droplets are combined after heating and quickly precipitated from the external polysaccharide network, which helps to improve the oral lubrication of the cooked food ( Figure 4 ).

[0057] Example 4

[0058] Preparation of chitosan-coated lipid nanoparticles: 3 parts by mass of chitosan and 7.5 parts by mass of Tween 40 were added to 100 mL of 2% w / v glacial acetic acid solution, and the mixture was placed in an 80°C water bath for 60 to 90 minutes until completely dissolved to obtain an aqueous phase. 10 parts by mass of beeswax was placed in an 80°C water bath for 20 to 35 minutes until completely melted to obtain an oil phase. The aqueous phase was then slowly poured into the oil phase at a similar temperature, stirred at 150 rpm for 5 minutes, sheared at a speed of 14,000 rpm with a high-speed shearing machine for 1 minute, and then ultrasonically treated at a power of 450 W for 7 minutes. The mixture was placed in a 4°C water bath, stirred, cooled for 1 hour, taken out, and then stored in a 4°C refrigerator for 24 hours for use to obtain chitosan-coated lipid nanoparticles.

[0059] Preparation of lipid-chitosan based Pickering high internal phase emulsion: The above particle dispersion was mixed with edible soybean oil in a volume ratio of 25:75, first treated with a high-speed shearing machine at 14000 rpm for 2 minutes, then rapidly cooled and stored in a refrigerator at 4°C for 24 hours to prepare an oil-in-water type lipid-chitosan based Pickering high internal phase emulsion.

[0060] Optical microscopic observation of the prepared lipid-chitosan based Pickering high internal phase emulsion: Freshly prepared samples were spotted on a glass slide using a capillary tube, and their morphology was observed using an optical microscope.

[0061] The prepared lipid-chitosan based Pickering high internal phase emulsion was subjected to constant temperature cooking test: the freshly prepared samples were heated at 70 °C in a convection oven and their melting behavior was recorded after 15 min.

[0062] Oral soft tribological evaluation of lipid-chitosan-based Pickering high internal phase emulsion cooking materials was performed: an MCR702e MultiDrive rheometer and a T-PTD200 ball tripod fitting were used to simulate oral soft friction to determine the lubrication properties of the cooking materials at 37°C. The simulated oral stress was 1.00 N and the sliding speed was 0.1-10 mm / s.

[0063] In Example 3, the food-grade lipid-chitosan-based Pickering high internal phase emulsion used a 1% w / v Tween solution. As the concentration of the emulsifier Tween increased to 1.875% w / v, the dispersion of the droplets of the high internal phase emulsion in this example was improved, giving the oil droplets a tighter connection, resulting in a stronger network structure of the emulsion and a more uniform overall texture ( Figure 1 ). When the temperature rises, Figure 2 The semi-solid shape of the high internal phase emulsion in this example could not be maintained after heating due to melting. Some liquid oil also flowed out around the commercial pork back fat cooking. It is worth noting that although the droplets in this example were uniform and small, the oral lubrication feeling was not significantly different from that of Example 3 ( Figure 4 ) This is because the small molecule emulsifier Tween 40 desorbs from the oil-water interface upon heating, accelerating the contact and aggregation of oil droplets. Overall, the lipid-chitosan-based Pickering high internal phase emulsions in Examples 1 to 4 have low saturated fat content and can facilitate the development of healthier and safer animal adipose tissue mimics.

[0064] Comparative Example 1

[0065] Preparation of casein-coated lipid nanoparticles: Add 7.5 parts by mass of casein to 100 mL of deionized water, place it in an 80°C water bath for 60 to 90 minutes until it is completely dissolved to obtain an aqueous phase, place 10 parts by mass of beeswax in an 80°C water bath for 20 to 35 minutes until it is completely melted to obtain an oil phase, then slowly pour the aqueous phase into the oil phase at a similar temperature, stir at 150 rpm for 5 minutes, shear at a speed of 14,000 rpm with a high-speed shearing machine for 1 minute, and then ultrasonically treat it at a power of 450 W for 7 minutes, place it in a 4°C water bath, stir and cool for 1 hour, take it out, and then store it in a 4°C refrigerator for 24 hours for use, to obtain casein-coated lipid nanoparticles.

[0066] Preparation of lipid-casein-based Pickering high internal phase emulsion: The above-mentioned particle dispersion was mixed with edible soybean oil in a volume ratio of 25:75, first treated with a high-speed shearing machine at 14000 rpm for 2 minutes, then rapidly cooled and stored in a refrigerator at 4°C for 24 hours before use, to obtain an oil-in-water type lipid-casein-based Pickering high internal phase emulsion.

[0067] Optical microscopic observation of the prepared lipid-casein based Pickering high internal phase emulsion: Freshly prepared samples were spotted on a glass slide using a capillary tube, and their morphology was observed using an optical microscope.

[0068] The lipid-casein based Pickering high internal phase emulsion was subjected to a constant temperature cooking test: the freshly prepared samples were heated at 70°C in a convection oven and their melting behavior was recorded after 15 min.

[0069] The rheological temperature scan of the prepared lipid-casein based Pickering high internal phase emulsion was performed: an MCR702e MultiDrive rheometer and a PP25 flat plate fixture were used to measure the thermal responsiveness of the emulsion in the temperature range of 4 to 80°C.

[0070] Oral soft tribological evaluation of the lipid-casein-based Pickering high internal phase emulsion cooking products was performed: the lubrication properties of the cooking products were determined at 37°C using an MCR702e MultiDrive rheometer and a T-PTD200 ball-and-pin fitting to simulate oral soft friction. The simulated oral stress was 1.00 N and the sliding speed was 0.1-10 mm / s.

[0071] The food-grade lipid-casein-based Pickering high internal phase emulsion of this comparative example was stored in a refrigerator at 4°C for 24 hours. The entire system was semi-solid and the droplet distribution was relatively uniform under optical microscope observation ( Figure 1 ). Figure 2 The high internal phase emulsion of the comparative example at room temperature is similar to the appearance of pig back fat, but Figure 3 The rheological scanning results showed that the external phase protein network was denatured by heat, which hindered the disintegration of the emulsion. After heating, the expected melting and oil separation performance was obviously lacking. Moreover, this would cause the oral lubricity of the cooked product of this comparative example to be far worse than that of pork back fat and Example 1 ( Figure 4 ).

[0072] Comparative Example 2

[0073] Preparation of chitosan-coated lipid nanoparticles: 3.5 parts by mass of chitosan and 4 parts by mass of Tween 40 were added to 100 mL of 2% w / v glacial acetic acid solution, and the mixture was placed in an 80°C water bath for 60 to 90 minutes until completely dissolved to obtain an aqueous phase. 15 parts by mass of beeswax was placed in an 80°C water bath for 20 to 35 minutes until completely melted to obtain an oil phase. The aqueous phase was then slowly poured into the oil phase at a similar temperature, stirred at 150 rpm for 5 minutes, sheared at a speed of 14,000 rpm with a high-speed shearing machine for 1 minute, and then ultrasonically treated at a power of 450 W for 7 minutes. The mixture was placed in a 4°C water bath, stirred, cooled for 1 hour, taken out, and then stored in a 4°C refrigerator for 24 hours for use to obtain chitosan-coated lipid nanoparticles.

[0074] Preparation of lipid-chitosan based Pickering high internal phase emulsion: The above particle dispersion was mixed with edible soybean oil in a volume ratio of 25:75, first treated with a high-speed shearing machine at 14000 rpm for 2 minutes, then rapidly cooled and stored in a refrigerator at 4°C for 24 hours to prepare an oil-in-water type lipid-chitosan based Pickering high internal phase emulsion.

[0075] Optical microscopic observation of the prepared lipid-chitosan based Pickering high internal phase emulsion: The freshly prepared sample was spotted on a glass slide using a capillary tube, and its morphology was observed using an optical microscope.

[0076] The food-grade lipid-chitosan-based Pickering high internal phase emulsion of this comparative example was placed in a refrigerator at 4°C for 24 hours. An unstable phase inversion occurred and the semi-solid state of the high internal phase emulsion could not be obtained. For details, please refer to Figure 1 Because excessively high levels of matrix beeswax will significantly increase the particle size, making it difficult for the high internal phase emulsion to be stable under the same shear conditions, it can be seen that a higher concentration of lipid nanoparticles does not necessarily lead to better stabilization of the high internal phase emulsion.

[0077] Comparative Example 3

[0078] Preparation of chitosan-coated lipid nanoparticles: 3.5 parts by mass of chitosan and 3 parts by mass of Tween 40 were added to 100 mL of 2% w / v glacial acetic acid solution, and the mixture was placed in an 80°C water bath for 60 to 90 minutes until completely dissolved to obtain an aqueous phase. 10 parts by mass of beeswax was placed in an 80°C water bath for 20 to 35 minutes until completely melted to obtain an oil phase. The aqueous phase was then slowly poured into the oil phase at a similar temperature, stirred at 150 rpm for 5 minutes, sheared at a speed of 14,000 rpm with a high-speed shearing machine for 1 minute, and then ultrasonically treated at a power of 450 W for 7 minutes. The mixture was placed in a 4°C water bath, stirred, cooled for 1 hour, taken out, and then stored in a 4°C refrigerator for 24 hours for use to obtain chitosan-coated lipid nanoparticles.

[0079] Preparation of lipid-chitosan based Pickering high internal phase emulsion: The above particle dispersion was mixed with edible soybean oil in a volume ratio of 25:75, first treated with a high-speed shearing machine at 14000 rpm for 2 minutes, then rapidly cooled and stored in a refrigerator at 4°C for 24 hours to prepare an oil-in-water type lipid-chitosan based Pickering high internal phase emulsion.

[0080] Optical microscopic observation of the prepared lipid-chitosan based Pickering high internal phase emulsion: Freshly prepared samples were spotted on a glass slide using a capillary tube, and their morphology was observed using an optical microscope.

[0081] Figure 1 The food-grade lipid-chitosan-based Pickering high internal phase emulsion in this comparative example underwent unstable phase inversion after being stored in a 4°C refrigerator for 24 hours, failing to achieve the semi-solid state of the high internal phase emulsion. Unlike Examples 3 and 4, this comparative example used a 0.75% w / v Tween solution. However, this lower emulsifier concentration had a less pronounced effect on particle modification, resulting in loose adsorption of the particles on the droplet surface, making the high internal phase emulsion difficult to stabilize.

[0082] Comparative Example 4

[0083] Preparation of chitosan-coated lipid nanoparticles: 4 parts by mass of chitosan and 4 parts by mass of Tween 40 were added to 100 mL of 2% w / v glacial acetic acid solution, and the mixture was placed in an 80°C water bath for 60 to 90 minutes until completely dissolved to obtain an aqueous phase. 10 parts by mass of beeswax was placed in an 80°C water bath for 20 to 35 minutes until completely melted to obtain an oil phase. The aqueous phase was then slowly poured into the oil phase at a similar temperature, stirred at 150 rpm for 5 minutes, sheared at a speed of 14,000 rpm with a high-speed shearing machine for 1 minute, and then ultrasonically treated at a power of 450 W for 7 minutes. The mixture was placed in a 4°C water bath, stirred, cooled for 1 hour, taken out, and then stored in a 4°C refrigerator for 24 hours for use to obtain chitosan-coated lipid nanoparticles.

[0084] Preparation of lipid-chitosan based Pickering high internal phase emulsion: The above particle dispersion was mixed with edible soybean oil in a volume ratio of 25:75, first treated with a high-speed shearing machine at 14000 rpm for 2 minutes, then rapidly cooled and stored in a refrigerator at 4°C for 24 hours to prepare an oil-in-water type lipid-chitosan based Pickering high internal phase emulsion.

[0085] Optical microscopic observation of the prepared lipid-chitosan based Pickering high internal phase emulsion: Freshly prepared samples were spotted on a glass slide using a capillary tube, and their morphology was observed using an optical microscope.

[0086] The results of Comparative Examples 2 and 3 are the same. The food-grade lipid-chitosan-based Pickering high internal phase emulsion of this comparative example was placed in a refrigerator at 4°C for 24 hours, and an unstable phase inversion occurred, and the semi-solid state of the high internal phase emulsion could not be presented ( Figure 1 The reason may be that the high chitosan concentration increases the viscosity of the water phase. Under the same shear conditions, the oil phase cannot be evenly mixed into the system, making the high internal phase emulsion unable to exist stably.

[0087] In summary, from Examples 1 to 4 and Comparative Examples 1 to 4, it can be seen that adjusting the properties of the interface layer by designing particles of chitosan, Tween and food-grade lipids will directly determine the stability of the water-in-oil lipid-chitosan-based Pickering high internal phase emulsion; due to the thermosensitivity of the particle matrix beeswax, the lipid-chitosan-based Pickering high internal phase emulsion will also exhibit different degrees of temperature response effects after heating, and can be used as a new type of fat simulated substance with hot melting behavior to enhance the taste appeal of bionic meat products.

[0088] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A method for preparing an animal adipose tissue mimic of a lipid-chitosan based Pickering high internal phase emulsion, characterized in that: The following steps are involved: (1) Chitosan and an emulsifier are heated and dissolved in a solvent to obtain an aqueous phase, and then a food-grade lipid is heated and melted to obtain an oil phase. The aqueous phase is added to the oil phase and stirred, followed by high-speed shearing, ultrasonic treatment, and cooling treatment to obtain a chitosan-coated lipid nanoparticle dispersion; (2) adding edible vegetable oil to the chitosan-coated lipid nanoparticle dispersion, and obtaining an animal adipose tissue mimic of the lipid-chitosan based Pickering high internal phase emulsion through high-speed shearing and rapid cooling; In step (1), the concentration of chitosan in the lipid-chitosan-based Pickering high internal phase emulsion is 0.5-0.9 g / 100 mL; the concentration of the emulsifier in the lipid-chitosan-based Pickering high internal phase emulsion is 1-2 g / 100 mL; the concentration of the food-grade lipid in the lipid-chitosan-based Pickering high internal phase emulsion is 1-3 g / 100 mL; The food-grade lipid in step (1) is a lipid with a melting point ≥42°C; In step (2), the volume ratio of the particle dispersion to the edible vegetable oil is 20-30:70-80.

2. The preparation method according to claim 1, characterized in that The emulsifier in step (1) is Tween; the solvent is a 1-3 g / 100 mL glacial acetic acid solution; the food-grade lipid is at least one of beeswax, palm stearin, and fully hydrogenated vegetable oil; and the edible vegetable oil is at least one of fish oil, corn oil, rapeseed oil, soybean oil, peanut oil, camellia oil, safflower oil, olive oil, walnut oil, sunflower seed oil, and linseed oil.

3. The preparation method according to claim 1, characterized in that The conditions for heating the water phase in step (1) are: temperature of 70-85° C., time of 60-90 min; the conditions for heating the oil phase are: temperature of 70-85° C., time of 20-35 min.

4. The preparation method according to claim 1, characterized in that The stirring in step (1) is magnetic stirring; the stirring conditions are: a rotation speed of 100 to 200 rpm and a time of 3 to 8 minutes.

5. The preparation method according to claim 1, characterized in that The high-speed shearing conditions in step (1) are: a rotation speed of 10,000 to 20,000 rpm, and a time of 1 to 5 minutes.

6. The preparation method according to claim 1, characterized in that The ultrasonic treatment conditions in step (1) are as follows: power of 100 to 500 W and time of 5 to 10 min.

7. The preparation method according to claim 1, characterized in that The cooling conditions in step (1) are as follows: temperature of 4 to 37° C. and time of ≥24 h.

8. The preparation method according to claim 1, characterized in that The high-speed shearing conditions in step (2) are: a rotation speed of 10,000 to 20,000 rpm, and a time of 2 to 5 minutes.

9. An animal adipose tissue mimetic of a lipid-chitosan based Pickering high internal phase emulsion, characterized in that: Prepared according to the preparation method according to any one of claims 1 to 8.

10. Use of the animal adipose tissue mimic of the lipid-chitosan based Pickering high internal phase emulsion according to claim 9 in the fields of food and / or medicine.

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