Composite embedded particle containing polylysine and continuous membrane emulsification preparation method thereof

Abstract

The invention provides a polylysine-containing composite embedded particle and a continuous membrane emulsification preparation method thereof, and the method comprises the following steps: dissolving epsilon-polylysine and galactose araban in water to obtain an internal water phase, mixing mixed MCT oil, phospholipid, polyglycerol fatty acid ester and hinokiol to obtain an external membrane oil phase, and carrying out continuous membrane emulsification on the external membrane oil phase to obtain the polylysine-containing composite embedded particle. Mixing the inner water phase and the outer membrane oil phase in a membrane emulsification mode to obtain W / O type primary emulsion; mixing polyglycerol fatty acid ester, cetostearyl alcohol, glycerol and water to obtain an out-of-membrane water phase, and mixing the W / O primary emulsion and the out-of-membrane water phase in a secondary membrane emulsification manner to obtain W / O / W multiple emulsion; the W / O / W multiple emulsion and maltodextrin are uniformly mixed in water and then spray-dried to obtain composite embedded particles containing polylysine, and stable encapsulation of epsilon-polylysine is realized by adopting an efficient and mild encapsulation technology.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of complexes, in particular to a polylysine-containing composite embedding particle and a continuous membrane emulsification preparation method thereof. BACKGROUND

[0002] As a natural antibacterial peptide, ε-polylysine (ε-PL) has a broad application prospect in the field of food preservation due to its safety, non-toxicity and good water solubility. It can effectively inhibit the growth and reproduction of various bacteria and provide reliable protection for food preservation. It has become a potential natural preservative in the food industry.

[0003] However, in practical application, the inherent characteristics of ε-polylysine pose many problems that need to be solved, which seriously limit the expansion of its application range and the exertion of its use effect. On the one hand, ε-polylysine has strong water solubility and cannot be uniformly dispersed when directly applied to high-fat food systems, resulting in uneven control of microorganisms in food and failure to fully exert its preservative effect. At the same time, ε-polylysine crystals or powders are prone to moisture absorption and caking, and the caked material is difficult to break again, which brings great inconvenience to production and processing, storage and transportation, and actual use, and increases production cost and operation difficulty.

[0004] On the other hand, the antibacterial spectrum of ε-polylysine is limited, and its inhibitory ability against fungi is weak. When used alone, it cannot meet the comprehensive prevention and control needs of various microorganisms (including bacteria and fungi) in complex food systems.

[0005] To solve the above problems, the industry attempts to modify ε-polylysine by coating technology to improve its dispersibility, stability and expand its antibacterial spectrum. However, when preparing the coated system by traditional homogenization emulsification method, long-time treatment is required at high temperature and high speed. This preparation method not only has the problems of high energy consumption and low efficiency, but also leads to large average particle size and uneven distribution of the coated system, high polydispersity index (PDI) and poor stability. More importantly, the high temperature and pressure treatment conditions can damage the heat-sensitive and light-sensitive auxiliary ingredients in the system, resulting in a decrease in their content and loss of activity, and can also cause structural changes in ε-polylysine itself, affecting its antibacterial effect. In addition, the existing coating technology cannot effectively coat oil-soluble ingredients with water-soluble ingredients, and cannot compensate for the defects of ε-polylysine antibacterial spectrum by compounding different active ingredients. During the coating process, the active ingredients are easily exposed and the coating rate is insufficient, so that the coated ε-polylysine is still prone to moisture absorption and adhesion during storage and use, which cannot fundamentally solve the application problems.

[0006] Therefore, the application of epsilon-polylysine on the market is limited, and there are many technical problems to be solved in the popularization and application of epsilon-polylysine. SUMMARY

[0007] The present application aims to provide a polylysine-containing composite embedding particle and a continuous membrane emulsification preparation method thereof, which realizes stable wrapping of epsilon-polylysine by using an efficient and mild wrapping technology, effectively improves the poor dispersibility of epsilon-polylysine in a high-fat system and improves the storage stability, and realizes the composite wrapping of oil-soluble antibacterial ingredients and water-soluble epsilon-polylysine to expand the antibacterial spectrum.

[0008] To achieve the above-mentioned purpose, the technical scheme provides a continuous membrane emulsification preparation method of a polylysine-containing composite embedding particle, which comprises the following steps: S1: preparing a W / O primary emulsion: dissolving epsilon-polylysine and galactoarabinan in water to obtain an inner water phase, mixing MCT oil, phospholipid, polyglycerol fatty acid ester and hinokitiol to obtain an outer oil phase of the membrane, and obtaining a W / O primary emulsion by using a membrane emulsification method; S2: preparing a W / O / W multiple emulsion: mixing polyglycerol fatty acid ester, cetyl stearyl alcohol, glycerol and water to obtain an outer water phase of the membrane, and mixing the W / O primary emulsion and the outer water phase of the membrane by using a secondary membrane emulsification method to obtain a W / O / W multiple emulsion; S3: preparing a polylysine-containing composite embedding particle: uniformly mixing the W / O / W multiple emulsion and maltodextrin in water, and then performing spray drying to obtain a polylysine-containing composite embedding particle.

[0009] The present scheme solves the problem of the difficulty of uniform dispersion of water-soluble epsilon-polylysine in a high-fat food by encapsulating the epsilon-polylysine in an oil phase system through a layered embedding technology of continuous membrane emulsification, realizes the uniformity of the control of microbial effects, avoids the moisture absorption and caking of polylysine crystals / powder, simplifies the production, storage and use processes, and the preparation conditions are more mild to minimize the color change and degradation of heat-sensitive ingredients (such as hinokitiol).

[0010] In step S1: epsilon-polylysine and galactoarabinan are used as the inner water phase, wherein the arabinan is used as a water-soluble polysaccharide, which can moderately increase the viscosity of the inner water phase when dissolved in water to avoid the loss or agglomeration of the inner water phase at the membrane holes, and the molecular chain of the galactoarabinan can form a mild protective film on the surface of the epsilon-polylysine molecules to prevent the inner water phase droplets from being damaged by shear force when passing through the membrane holes, thereby ensuring that the epsilon-polylysine is completely wrapped into the oil phase and laying a foundation for the subsequent high encapsulation rate.

[0011] In some embodiments, the galactoarabinan is a mixture of galactose and arabinan, and the molar ratio of galactose to arabinan is 6:1.

[0012] In some embodiments, 8% ε-polylysine and 1% galactoarabinogalactan are added to 91% deionized water.

[0013] Furthermore, this scheme uses a mixed solution of MCT oil, phospholipids, polyglycerol fatty acid esters, and pine phenol as the outer oil phase of the membrane. The emulsifying synergistic effect of the mixed MCT oil with phospholipids and polyglycerol fatty acid esters can reduce the oil-water interfacial tension, allowing the inner aqueous phase to form uniform droplets when passing through the hydrophobic membrane. Phospholipids and polyglycerol fatty acid esters act as composite emulsifiers to form a stable membrane structure at the oil-water interface, reducing the leakage of ε-polylysine.

[0014] In addition, as mentioned earlier, ε-polylysine has a poor inhibitory effect on fungi. Therefore, this solution adds physalisol to form an "oil-water phase" synergistic antibacterial system with polylysine in the inner aqueous phase (which has an antibacterial spectrum that focuses on bacteria). This is to make up for the weakness of polylysine in inhibiting fungi and improve the overall microbial control effect of the product in food.

[0015] In some embodiments, the mixed MCT oil uses a C8 to C10 ratio of 6:4, meaning the mixed MCT oil consists of 6 parts by mass of C8 (octanoic acid) and 4 parts by mass of C10 (decanoic acid). Since MCT oil is a saturated fatty acid extracted and processed from coconut oil or palm kernel oil, primarily composed of C6, C8, C10, and C12 triglycerides, its viscosity increases with the carbon chain. For membrane emulsification, choosing a 6:4 combination of C8 and C10 with lower viscosity is crucial. This is because, for example, pure C6 has too low a viscosity, resulting in excessively fine emulsion particles, introducing more surfactants, and a lower loading of active ingredients. Conversely, C12 has too high a viscosity, requiring excessively high gas pressure, which is detrimental to production and equipment maintenance. The 6:4 ratio of C8 to C10 depends on the required gas pressure, the type of emulsifier, and the resulting emulsion particle size. It is also a relatively energy-efficient choice, and the cost of the mixed oil is lower than that of pure oil.

[0016] In some embodiments, 50-90% by weight of mixed MCT oil, 1-10% by weight of phospholipids, 2-12% by weight of polyglycerol fatty acid esters and 5-12% by weight of pine phenol are mixed to obtain an external oil phase, and the internal aqueous phase and the external oil phase are mixed by membrane emulsification to obtain a W / O type primary emulsion.

[0017] Preferably, an extracellular oil phase is obtained by mixing 79% MCT oil, 3% phospholipids, 8% polyglycerol fatty acid esters and 10% pine phenol by volume.

[0018] Furthermore, the phospholipid is selected as any or a combination of lecithin, soy lecithin, etc.

[0019] Furthermore, the polyglycerol fatty acid value is selected as any or a combination of polyglycerol-10-laurate and polyglycerol-10-myristate.

[0020] It should be noted that this scheme uses homogenization to separately prepare the internal aqueous phase and the external oil phase. Furthermore, this scheme uses membrane emulsification to mix the internal aqueous phase and the external oil phase to obtain a W / O type primary emulsion. A schematic diagram of the preparation process for the W / O type primary emulsion is shown below. Figure 2 As shown.

[0021] In some embodiments, a hydrophobic membrane with a specific pore size is installed in a membrane emulsifier, the inner aqueous phase is placed inside the hydrophobic membrane, the outer oil phase is placed outside the hydrophobic membrane, and an inert gas is introduced into the inner aqueous phase side to push the inner aqueous phase through the hydrophobic membrane and disperse it into the outer oil phase to form a W / O type primary emulsion.

[0022] Furthermore, a 0.2 μm hydrophobic membrane is installed inside the membrane emulsifier. This approach selects a 0.2 μm hydrophobic membrane to strictly limit the size of droplets formed in the internal aqueous phase, avoiding excessively large or uneven droplets. This results in a minimum average particle size of 458 nm for the primary emulsion and a polydispersity index (PDI) as low as 0.12, significantly better than the particle size dispersion effect of traditional homogenization, providing high-quality intermediate products for subsequent secondary membrane emulsification.

[0023] In some embodiments, the hydrophobic membrane is selected as a silanized modified SPG membrane or a polymer membrane. Further, the material of the hydrophobic membrane is selected from one or more of the following: polyvinylidene fluoride, polytetrafluoroethylene, polypropylene, polyethylene, and siloxane-modified ceramics.

[0024] In some embodiments, nitrogen gas is introduced into the aqueous phase side to control the pressure at 30-100 kPa. Preferably, the control pressure is 80-85 kPa.

[0025] In some embodiments, the flow rate of the extracellular oil phase is controlled at 50~150 mL / min, and the temperature is controlled at 25~32℃.

[0026] It should be noted that when the flow rate of the oil phase outside the membrane is too low, droplets formed by the water phase penetrating the membrane pores are prone to agglomeration and merging on the membrane surface, leading to larger particle size, poorer particle size uniformity, and increased PDI. Conversely, excessively high flow rates will generate excessive turbulence, disrupting the newly formed micro-droplets and similarly affecting uniformity. In other words, an appropriate flow rate allows phospholipids and polyglycerol fatty acid esters in the oil phase to quickly form a complete emulsion film at the water-oil interface, reducing leakage of the water phase.

[0027] In addition, temperature control of the outer oil phase can moderately reduce the viscosity of the mixed MCT oil, improve the oil phase fluidity, and weaken the water-oil interfacial tension, making it easier for the inner aqueous phase to penetrate the hydrophobic membrane pores to form tiny droplets, reducing the pressure required for emulsification and avoiding a sharp drop in emulsification efficiency due to excessive viscosity.

[0028] Preferably, the flow rate of the oil phase outside the membrane is controlled at 100 mL / min, and the temperature is controlled at 28~29℃.

[0029] In some embodiments, the temperature of the internal aqueous phase is controlled at 27~29°C. Preferably, the temperature of the internal aqueous phase is controlled at 28°C.

[0030] In some embodiments, the ratio of the internal aqueous phase to the external oil phase is 1:(1~3).

[0031] Preferably, the ratio of the internal aqueous phase to the external oil phase is 1:1.2.

[0032] In step S2: Step S1 of this scheme yields a W / O type colostrum. Although the inner aqueous phase has been initially encapsulated by an external oil phase, if directly applied to a water-containing food system (such as beverages, sauces, etc.), it is prone to layering and aggregation due to oil-water immiscibility, making uniform dispersion impossible. Furthermore, the ε-polylysine in the inner aqueous phase may still leak due to oil phase damage, and the W / O type colostrum has weak resistance to moisture absorption and external interference. Therefore, this scheme further employs a secondary membrane emulsification using an external oil phase to encapsulate the W / O type colostrum, forming a W / O / W double-layer encapsulation structure, namely, an encapsulation structure consisting of an external aqueous phase, an external oil phase, and an inner aqueous phase.

[0033] In some embodiments, a mixture of 5-25% polyglycerol fatty acid ester, 1-10% cetearyl alcohol, 1-10% glycerol and 50-90% water by volume is obtained to obtain an extramembrane aqueous phase.

[0034] Polyglycerol fatty acids, as key emulsifiers, possess both hydrophilic and lipophilic groups, which can reduce the interfacial tension between the outer oil phase and the outer aqueous phase of the membrane. At the same time, this component works synergistically with the emulsifier in the outer oil phase of the W / O primary emulsion to induce the formation of a stable liquid crystal "mesh" structure in the re-emulsion particles, firmly locking the internal oil phase and the internal aqueous phase, preventing the leakage of active ingredients, and laying the foundation for high encapsulation efficiency and long-term stability.

[0035] Cetearyl alcohol, as a co-emulsifier, can moderately increase the viscosity of the aqueous phase outside the membrane, preventing the primary oil droplets from agglomerating or settling too quickly during secondary membrane emulsification. At the same time, its molecules can insert into the interfacial membrane formed by polyglycerol fatty acid esters, strengthening the membrane structure, reducing oil droplet breakage during subsequent spray drying, and improving the integrity of the encapsulated particles.

[0036] Glycerin has strong moisturizing properties, which can prevent excessive water loss of the aqueous phase outside the membrane during the process, thus preventing the system from becoming viscous and ensuring the fluidity of the membrane during emulsification. At the same time, its water-soluble properties can improve the compatibility of polyglycerol fatty acid esters, cetearyl alcohol and water, making the aqueous phase outside the membrane more uniformly mixed, avoiding excessively high or low concentrations of emulsifier in some areas, and ensuring stable secondary emulsification effect.

[0037] Preferably, an extracellular aqueous phase is obtained by mixing 8% polyglycerol fatty acid ester, 2% cetearyl alcohol, 3% glycerol and 87% water by volume.

[0038] In some embodiments, the types of polyglycerol fatty acids include at least one or more of the following: PEG-4 polyglycerol-2 distearate, PEG-4 polyglycerol-2 stearate, PPG-14 polyglycerol-2 ether, PPG-70 polyglycerol-10 ether, PPG-9 dipolyglycerol ether, polyglycerol-10 dimyristate, polyglycerol-10 eicosodecarboxylate / tetradecanoic acid ester, polyglycerol-10 eicosodecarboxylate / tetradecanoic acid esters, polyglycerol-10 diisostearate, polyglycerol-10 distearate, polyglycerol-10 dioleate, polyglycerol-10 dilaurate, polyglycerol-10 dipalmitate, polyglycerol-10 myristate, polyglycerol-10 tristearate, polyglycerol-10 trioleate, polyglycerol-10 isostearate, polyglycerol-10 stearate, polyglycerol-10 oleate, polyglycerol-10 laurate, polyglycerol-2 sesquicaprate, polyglycerol-2 Dimeric hydroxystearate, polyglycerol-2 diisostearate, polyglycerol-2 diisostearate / IPDI copolymer, polyglycerol-2 distearate, polyglycerol-2 dioleate, polyglycerol-2 decanoate, polyglycerol-2 isopalmitate, polyglycerol-2 stearate, polyglycerol-2 oleate, polyglycerol-2 lauryl ester, polyglycerol-3 diisostearate, polyglycerol-3 distearate, polyglycerol-3 beeswax ester, polyglycerol-3 decanoate, polyglycerol-3 methylglucose diisostearate, polyglycerol-3 methylglucose distearate, polyglycerol-3 cetyl ether, polyglycerol-3 polyricinoleate, polyglycerol-3 hydroxylaurate, polyglycerol-3 myristate, polyglycerol-3 pentaricinoleate, polyglycerol-3 caprylate, polyglycerol-3 cocoate, polyglycerol-3 isostearate, polyglycerol-3 stearate, polyglycerol-3 Oleate, Polyglycerol-3 Laurate, Polyglycerol-3 Palmitate, Polyglycerol-4 Diisostearate / Polyhydroxystearate / Sebacic Acid, Polyglycerol-4 Dilaurate, Polyglycerol-4 Decanoate, Polyglycerol-4 Isostearate, Polyglycerol-4 Stearate, Polyglycerol-4 Oil Ether, Polyglycerol-4 Oleate, Polyglycerol-4 Lauryl Ether, Polyglycerol-4 Laurate, Polyglycerol-5 Polyricinoleate, Polyglycerol-5 Myristate, Polyglycerol-5 Trioleate, Polyglycerol-5 Isostearate, Polyglycerol-5 Stearate, Polyglycerol-5 Oleate, Polyglycerol-5 Laurate, Polyglycerol-6 Octostearate, Polyglycerol-6 Ricinoleate, Polyglycerol-6 Didecanoate, Polyglycerol-6 Distearate, Polyglycerol-6 Dioleate, Polyglycerol-6 Polyricinoleate, Polyglycerol-6 Polyhydroxystearate, Polyglycerol-6 Myristate, polyglycerol-6 tristearate, polyglycerol-6 behenate, polyglycerol-6 pentastearate, polyglycerol-6 caprylate, polyglycerol-6 isostearate, polyglycerol-6 stearate, polyglycerol-6Oleate, polyglycerol-6 lauryl ester, polyglycerol-8 decaerucic acid ester / decaisostearate / decaricinoleate, polyglycerol-8 stearate, polyglycerol-8 stearate, polyglycerol sorbitol.

[0039] The types of cetearyl alcohol include at least one or more of the following: cetearyl alcohol, isocetearyl alcohol, phytosterol / isostearyl alcohol / cetearyl alcohol / stearyl / behenol di-linoleate, cetearyl alcohol, isostearyl alcohol, stearyl alcohol, behenol, phytosterol / behenol / octyldodecyl lauroyl glutamate, palmitic acid, stearic acid, phytosterols, brassosterols, daidzeinols, lanosterol, tallol, dihydrolanosterol, rice bran sterol, cholesterol, β-sitosterol, 7-dehydrocholesterol, and cetearyl alcohol polyether.

[0040] Similarly, step S2 of this scheme still uses a secondary membrane emulsification method to mix the W / O primary emulsion and the aqueous phase outside the membrane to obtain a W / O / W composite emulsion. Specifically, this scheme uses a secondary membrane emulsification method to mix the aqueous phase outside the membrane and the W / O primary emulsion to obtain a W / O / W composite emulsion. A schematic diagram of the preparation process of the W / O / W composite emulsion is shown below. Figure 3 As shown.

[0041] In some embodiments, a hydrophilic membrane is installed inside the membrane emulsifier, and the aqueous phase outside the membrane and the W / O pre-emulsion are placed on opposite sides of the hydrophilic membrane. An inert gas is introduced into the W / O pre-emulsion side to push the W / O pre-emulsion through the hydrophilic membrane and disperse it into the aqueous phase outside the membrane to form a W / O / W double emulsion.

[0042] In some embodiments, the hydrophilic membrane is selected as a 1μm hydrophilic membrane. The surface of the hydrophilic membrane is compatible with the aqueous phase outside the membrane, allowing the aqueous phase outside the membrane to adhere to the surface of the hydrophilic membrane. At the same time, the 1μm pore size precisely matches the dispersion requirements of W / O colostrum. This allows the W / O colostrum to penetrate the membrane pores under pressure and disperse into uniform droplets that are encapsulated by the aqueous phase outside the membrane. It also avoids the colostrum clogging the membrane pores due to excessively small pore size, or droplet aggregation due to excessively large pore size. Ultimately, a complete W / O / W structure of "aqueous phase outside the membrane - oil phase - aqueous phase inside the membrane" is formed, and composite emulsions containing multinucleated water-in-oil microparticles can be prepared in a directional manner.

[0043] In some embodiments, the hydrophilic membrane is selected as an SPG membrane or a ceramic membrane.

[0044] Furthermore, the material of the hydrophilic membrane is selected from at least one or more of the following types: alumina, zirconium oxide, titanium dioxide, hydrophilic polyvinylidene fluoride, hydrophilic polyethersulfone, cellulose, polyamide, and metal.

[0045] In some embodiments, nitrogen gas is introduced into the W / O type colostrum side to control the pressure at 10-30 kPa. Preferably, the control pressure is 15 kPa. The W / O type colostrum has a relatively small average particle size, and the low pressure of 10-30 kPa is just enough to push the colostrum through the hydrophilic membrane pores and disperse it into uniform droplets. If the pressure is too high, the colostrum particles will pass through the membrane pores at high speed and high shear, easily breaking and leaking active ingredients. If the pressure is too low, it will not be able to effectively penetrate the membrane pores or the emulsification efficiency will be low, and the re-emulsion particles will be too large and easily break.

[0046] In some embodiments, the flow rate of the aqueous phase outside the membrane is controlled at 10~50 mL / min, and the temperature is controlled at 27~29°C. Similarly, if the flow rate is too low, the aqueous phase outside the membrane will be replaced slowly, the emulsification efficiency will be low, and the W / O primary emulsion will easily aggregate on the membrane surface after penetrating the pores of the hydrophilic membrane; if the flow rate is too high, turbulence will be generated, which will destroy the newly formed droplets.

[0047] Preferably, the flow rate of the aqueous phase outside the membrane is controlled at 30 mL / min, and the temperature is controlled at 28 °C.

[0048] In some embodiments, the temperature of the W / O type colostrum is controlled at 25~32°C. Preferably, the temperature of the oil phase (colostrum) in the membrane is controlled at 28~29°C.

[0049] In some embodiments, the ratio of W / O type colostrum to extracellular aqueous phase is 1:(1~10).

[0050] Preferably, the ratio of W / O type colostrum to extracellular aqueous phase is 1:3.

[0051] In step S3, the W / O / W double emulsion prepared in step S2 of this scheme is then spray-granulated to obtain composite encapsulated particles containing polylysine.

[0052] In some embodiments, maltodextrin is added to a W / O / W double emulsion and uniformly dispersed to obtain a mixture. The mixture is then spray-granulated and sieved to obtain polylysine-containing composite encapsulated particles.

[0053] In some embodiments, 5-25% maltodextrin is added to the W / O / W double emulsion and uniformly dispersed by mechanical dispersion to obtain a mixture.

[0054] Because maltodextrin (DE value around 15) has good film-forming ability, it can quickly encapsulate W / O / W double emulsion droplets during spray drying, forming a dense solid shell that firmly locks in the internal ε-polylysine (soluble in the inner aqueous phase) and physalisol (soluble in the oil phase), preventing the active ingredients from leaking or degrading during high-temperature drying. At the same time, it forms regular particles, which are convenient for subsequent applications.

[0055] Preferably, 12.5% ​​maltodextrin is added to the W / O / W double emulsion and dispersed at 200 r / min for half an hour to obtain a mixture.

[0056] In some embodiments, the conditions for spray granulation are an inlet air temperature of 140~170°C and an outlet air temperature of 60~80°C.

[0057] In some embodiments, the particle size of the polylysine-containing composite encapsulated particles prepared is 100-300 mesh.

[0058] Secondly, this solution provides a composite encapsulated particle containing polylysine, comprising: The maltodextrin shell and the composite particles encapsulated within the maltodextrin shell; wherein the composite particles are a composite structure consisting of multiple W / O droplets encapsulated by an external aqueous phase, and the internal aqueous phase of the W / O droplets contains polylysine.

[0059] In some embodiments, the W / O droplet is a droplet formed by an aqueous phase and an oil phase, wherein the aqueous phase contains polylysine and galactoarabinogalactan, and the oil phase includes MCT oil, phospholipids, polyglycerol fatty acid esters, and cypressin.

[0060] In some embodiments, the outer aqueous phase of a W / O / W double emulsion comprises polyglycerol fatty acid ester, cetearyl alcohol, glycerol, and water.

[0061] In some embodiments, a polyglycerol mediated liquid crystal mesh structure is formed between the outer aqueous phase of the W / O / W type double emulsion and the W / O type primary emulsion to further enhance encapsulation stability and prevent leakage or reaction of the phase components.

[0062] In other words, the composite particles are multi-core oil-in-water microparticles formed through secondary continuous membrane emulsification; that is, the composite particles consist of multiple W / O droplets encapsulated by the external aqueous phase. A schematic diagram of the polylysine-containing composite encapsulated particles in this scheme is shown below. Figure 4 As shown.

[0063] Compared with existing technologies, this technical solution has the following characteristics and beneficial effects: This solution employs a continuous membrane emulsification technology that combines primary hydrophobic membrane emulsification with secondary hydrophilic membrane emulsification. By precisely controlling parameters such as membrane pore size, nitrogen pressure, phase flow rate, and temperature, it directionally prepares W / O / W double emulsions containing multinucleated water-in-oil (W / O) microparticles. This allows for precise control of the emulsification process and product particle size, resulting in higher repeatability and better suitability for large-scale production compared to traditional homogenization processes. Furthermore, in terms of composition, the inner aqueous phase of this solution uses ε-polylysine combined with galactosarabinogalactan, while the outer oil phase comprises a mixture of MCT oil, phospholipids, polyglycerol fatty acid esters, and physalisol. The outer aqueous phase is further combined with polyglycerol fatty acid esters and cetearyl alcohol to form a stable system. Finally, the mixture is spray-dried with maltodextrin to form a shell, achieving highly efficient composite encapsulation of water-soluble and oil-soluble active ingredients. Attached Figure Description

[0064] Figure 1 This is a schematic flowchart of the continuous film emulsification preparation method for polylysine-containing composite embedded particles according to this scheme.

[0065] Figure 2 This is a schematic diagram of the preparation process for W / O type colostrum.

[0066] Figure 3 This is a schematic diagram of the preparation process for W / O / W double emulsion.

[0067] Figure 4 This is a schematic diagram of the structure of polylysine-containing composite encapsulated particles.

[0068] Figure 5 This is an optical polarizing microscope liquid crystal image of Example 10 on the 7th day after secondary film emulsification.

[0069] Figure 6 This is an optical polarizing microscope liquid crystal image of Example 18 taken on the 7th day after secondary film emulsification.

[0070] Figure 7 This is a microscopic observation image of the polylysine-containing composite embedded particles of this scheme. Detailed Implementation

[0071] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention are within the scope of protection of the present invention.

[0072] I. Examples and comparative examples of W / O type colostrum: Example 1 An inner aqueous phase was obtained by dissolving 8% ε-polylysine and 1% galactosarabinogalactan in 91% deionized water. An outer oil phase was obtained by mixing 79% of a C8 and C10 (6:4) mixed MCT oil, 3% lecithin, 8% polyglycerol-10-laurate, and 10% pine phenol. 0.2 μm silanized modified SPG membranes were installed at intervals in a membrane emulsifier as hydrophobic membranes. The inner aqueous phase was placed inside the spaced-apart hydrophobic membranes, and the outer oil phase was placed outside the hydrophobic membranes. An inert gas was introduced into the inner aqueous phase to push it through the hydrophobic membranes and disperse it into the outer oil phase to form a W / O type primary emulsion. The pressure of the inert gas was controlled at 32.5 kPa, the flow rate of the outer oil phase outside the hydrophobic membrane was 50 mL / min, the temperature of the outer oil phase outside the hydrophobic membrane was 25.5 °C, and the temperature of the inner aqueous phase inside the hydrophobic membrane was 28 °C.

[0073] Examples 2 to 9: To verify the effect of different homogenization conditions on the formed W / O type colostrum, the applicant adjusted the specific homogenization conditions to obtain Examples 2 to 9. The homogenization conditions of Examples 2 to 9 are shown in Table 1 below. Table 1. Homogenization conditions for Examples 2 to 9

[0074] The performance test results of the W / O colostrum obtained in Examples 1 to 9 are shown in Table 2. Table 2: Indicator Results of Examples 1 to 9 .

[0075] The "average particle size" in the index was obtained by wet laser diffraction, with pure water as the dispersion medium, a temperature of 25.0℃, a duration of 60s, a material refractive index of 1.59, a medium refractive index of 1.33, and a material absorptivity of 0.010. Regarding the "ultrafiltration encapsulation rate" in the index, the ε-polylysine content was determined by ultrafiltration centrifugation: 1 mL of W / O colostrum was added to ultrafiltration centrifuge tubes with molecular weight cutoffs of 30 kD, 50 kD, and 100 kD. The tubes were centrifuged at a low speed (4000 r / min) for 10 min. The filtrate was periodically collected for high performance liquid chromatography to determine the ε-polylysine content.

[0076] High performance liquid chromatography method conditions: Liquid Chromatography (HPLC): Diode array detector; Column: C18 (4.6 mm × 250 mm, 5 µm); Mobile phase: Phosphate buffer: Acetonitrile = 92:8; Flow rate: 0.5 mL / min; Detection wavelength: 210 nm The "polydispersity index (PDI)" in the index was obtained by wet laser diffraction with pure water as the dispersion medium, at a temperature of 25.0℃, for a duration of 60s. The material refractive index was 1.59, the medium refractive index was 1.33, and the material absorptivity was 0.010.

[0077] In addition, Table 2 shows that the PDI index monitoring after 21 days is used to illustrate the stability of the emulsion particles. The smaller the PDI, the more uniform the emulsion; the smaller the change in PDI between the two time points, the more stable the emulsion.

[0078] Comparative Examples 1 to 9: The inner aqueous phase was obtained by dissolving 8% ε-polylysine and 1% galactoarabinogalactan in 91% deionized water. The outer oil phase was obtained by mixing 79% of a C8 and C10 (6:4) mixed MCT oil, 3% lecithin, 8% polyglycerol-10-laurate, and 10% pine phenol. The inner aqueous phase and the outer oil phase were homogenized using a traditional homogenization method, which involved shearing and emulsifying the aqueous and oil phases at a high pressure homogenizer of 5000-12000 r / min to prepare the primary emulsion. Comparative Examples 1 to 9 were obtained by adjusting the homogenization conditions. The homogenization conditions and corresponding primary emulsion indices of Comparative Examples 1 to 9 are shown in Table 3 below. Table 3. Homogenization conditions and corresponding colostrum indices for Comparative Examples 1 to 9

[0079] As shown in Table 3, increasing the homogenization temperature significantly improves emulsification; insufficient temperature greatly reduces emulsification efficiency. However, higher temperatures are not favorable for heat-sensitive active ingredients (such as physalisol), potentially leading to discoloration and degradation. Furthermore, the temperature typically increases with prolonged homogenization time, making precise control difficult, and higher temperatures also increase the risk of active ingredient degradation. Simultaneously, the quality of emulsions prepared using this process, including particle size uniformity, is not ideal.

[0080] In addition, the content of physalisol in the W / O type colostrum of Examples 1 to 9 and Comparative Examples 1 to 9 was detected by high performance liquid chromatography (HPLC), and the results are shown in Table 4. The HPLC detection conditions were as follows: Liquid Chromatography (HPLC): Diode array detector; Column: C18 (4.6 mm × 250 mm, 5 µm); Mobile phase: Acetonitrile: Water: Trifluoroacetic acid (500:500:0.5); Flow rate: 1 mL / min; Detection wavelength: 310 nm Table 4. Results of High Performance Liquid Chromatography (HPLC) Detection

[0081] As shown in Table 4, physalisol dissolves in the oil phase. During high-pressure shear emulsification using a homogenizer, the temperature gradually increases to over 50 degrees Celsius. This high temperature poses a risk of degradation and loss to heat-sensitive active ingredients such as physalisol. Membrane emulsification allows for precise temperature control and is a gentler, lower-temperature process, which is beneficial for the stability of heat-sensitive or photosensitive components. Data shows that membrane emulsification better maintains the stability of active ingredients and has a lower attenuation rate.

[0082] II. Examples and Comparative Examples of W / O / W Composite Emulsions: Examples 10 to 18: The W / O type primary emulsion obtained in Example 8 was mixed with 6% polyglycerol-6-distearate, 2% polyglycerol-10-stearate, 2% cetearyl alcohol, 3% glycerol, and 87% water to obtain an extra-membrane aqueous phase. A 1 μm SPG membrane was installed in a membrane emulsifier as a hydrophilic membrane. The extra-membrane aqueous phase and the W / O primary emulsion were placed on opposite sides of the hydrophilic membrane. An inert gas was introduced through the W / O primary emulsion side to push the W / O primary emulsion through the hydrophilic membrane and disperse it into the extra-membrane aqueous phase to form a W / O / W complex emulsion. The preparation conditions and corresponding index results of Examples 10 to 18 are shown in Table 5 below: Table 5. Preparation conditions and corresponding index results for Examples 10-18

[0083] Table 5 illustrates the initial PDI index and the PDI index after 21 days. A lower PDI value indicates better particle size uniformity, and a smaller change in PDI indicates less change in particle size uniformity and better stability. Additionally, the average particle size should not be too large, as excessively large particle size will reduce the stability of the emulsion particles.

[0084] In addition, the W / O / W composite emulsions prepared in Examples 10 and 18 were observed under an optical polarizing microscope on the 7th day, and the W / O / W composite emulsion corresponding to Example 10 was obtained as follows: Figure 5 As shown, the W / O / W composite emulsion corresponding to Example 18 is as follows: Figure 6 As shown, the arrangement of different emulsifiers on the surface of emulsion particles forms an intermediate phase between liquid and solid, known as liquid crystal. Liquid crystal emulsion particles display a "Maltese cross" pattern under a polarized light microscope, caused by birefringence.

[0085] Comparative Examples 10 to 18: The W / O type primary emulsion prepared in Comparative Example 6 was taken and mixed with 6% polyglycerol-6-distearate, 2% polyglycerol-10-stearate, 2% cetearyl alcohol, 3% glycerol and 87% water to obtain an extra-membrane aqueous phase. The extra-membrane aqueous phase and the W / O type primary emulsion were homogenized at a ratio of 3:1. The rotation speed, temperature and time were adjusted to monitor the technical indicators of re-emulsification. The preparation conditions and corresponding index results of Comparative Examples 10 to 18 are shown in Table 6 below: Table 6. Preparation conditions and corresponding index results for Comparative Examples 10 to 18

[0086] From the perspectives of average particle size and polydispersity index, secondary homogenization emulsification can only partially achieve W / O / W type encapsulation, rarely forming complex emulsions containing multiple water-in-oil particles. A smaller average particle size indicates that some water-in-oil mononuclear particles are re-encapsulated by the emulsifier, forming W / O / W type encapsulations, but with less oil phase content. A poor polydispersity index indicates that some oil phase is directly encapsulated within the emulsifier shell; these encapsulations lack an internal aqueous phase containing polylysine, leading to uneven active ingredient content during spray drying. Some primary emulsion particles break down under homogenization shear, and the internal aqueous phase containing polylysine dissolves into the aqueous phase during secondary emulsification and participates in the emulsification process. In this case, polylysine appears in the encapsulation shell, easily adhering upon moisture, and the exposed active ingredient lacks the encapsulation advantage.

[0087] III. Spray granulation: Maltodextrin (DE value of approximately 15) was added at a ratio of 12.5% ​​to the W / O / W type double emulsion of Example 14. The mixture was mechanically dispersed at 200 rpm for half an hour until uniformly dissolved, and then spray-dried. The preferred spray-drying conditions were an inlet air temperature of 140-170℃ and an outlet air temperature of 60-80℃. The mixture was sieved to obtain composite encapsulated shell particles with a 100-300 mesh maltodextrin outer shell. The micrograph is shown below. Figure 7 As shown.

[0088] Those skilled in the art should understand that the technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments have been described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0089] The above embodiments are merely illustrative of several implementation methods of this application, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of this application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this application should be determined by the appended claims.

Claims

1. A continuous film emulsification preparation method for composite encapsulated particles containing polylysine, characterized in that, Includes the following steps: S1: Preparation of W / O type colostrum: ε-polylysine and galactoarabinogalactan are dissolved in water to obtain an inner aqueous phase. Mixed MCT oil, phospholipids, polyglycerol fatty acid esters and pine phenol are mixed to obtain an outer oil phase. W / O type colostrum is obtained by membrane emulsification. S2: Preparation of W / O / W double emulsion: Mix polyglycerol fatty acid ester, cetearyl alcohol, glycerol and water to obtain an aqueous phase outside the membrane. Mix the W / O primary emulsion and the aqueous phase outside the membrane using a two-stage membrane emulsification method to obtain a W / O / W double emulsion. S3: Preparation of polylysine-containing composite encapsulated particles: W / O / W double emulsion and maltodextrin were mixed evenly in water and then spray-dried to obtain polylysine-containing composite encapsulated particles.

2. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 1, characterized in that, The MCT oil is selected with a mass ratio of C8 and C10 of 6:

4.

3. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 1, characterized in that, The phospholipid is selected as any or a combination of lecithin and soybean lecithin; the polyglycerol fatty acid value is selected as any or a combination of polyglycerol-10-laurate, polyglycerol-6-distearate, and polyglycerol-10-stearate.

4. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 1, characterized in that, A hydrophobic membrane with a specific pore size is installed inside a membrane emulsifier. The inner aqueous phase is placed inside the hydrophobic membrane, and the outer oil phase is placed outside the hydrophobic membrane. An inert gas is introduced into the inner aqueous phase side to push the inner aqueous phase through the hydrophobic membrane and disperse it into the outer oil phase to form a W / O type primary emulsion.

5. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 4, characterized in that, A 0.2 μm hydrophobic membrane is installed in the membrane emulsifier. Nitrogen gas is introduced into the inner aqueous phase side to control the pressure at 30~100 kPa. The flow rate of the outer oil phase is controlled at 50~150 mL / min, and the temperature of the inner aqueous phase is controlled at 27~29℃.

6. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 1, characterized in that, The ratio of the internal aqueous phase to the external oil phase is 1:(1~3), and the ratio of the W / O type colostrum to the external aqueous phase is 1:(1~10).

7. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 1, characterized in that, A hydrophilic membrane is installed inside the membrane emulsifier. The aqueous phase outside the membrane and the W / O pre-emulsion are placed on opposite sides of the hydrophilic membrane. An inert gas is introduced into the W / O pre-emulsion side to push the W / O pre-emulsion through the hydrophilic membrane and disperse it into the aqueous phase outside the membrane to form a W / O / W double emulsion.

8. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 7, characterized in that, A 1μm hydrophilic membrane was selected. Nitrogen gas was introduced into the W / O type colostrum side to control the pressure at 10~30KPa. The flow rate of the aqueous phase outside the membrane was controlled at 10~50mL / min. The temperature of the W / O type colostrum was controlled at 25~32℃.

9. The continuous film emulsification preparation method of polylysine-containing composite embedded particles according to claim 1, characterized in that, Mixing 50-90% by weight of mixed MCT oil, 1-10% by weight of phospholipids, 2-12% by weight of polyglycerol fatty acid esters and 5-12% by weight of pine phenols yields an external oil phase. The internal aqueous phase and the external oil phase are then mixed using membrane emulsification to obtain a W / O type primary emulsion.

10. A composite encapsulated particle containing polylysine, characterized in that, The composite encapsulated particles containing polylysine are prepared by a continuous membrane emulsification method according to any one of claims 1 to 9, comprising: a maltodextrin shell and composite particles encapsulated within the maltodextrin shell; wherein the composite particles are a composite structure consisting of multiple W / O droplets encapsulated by an external aqueous phase, and the internal aqueous phase of the W / O droplets contains polylysine.

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