Preparation method of micro-nano multi-compartment polysaccharide colloid material

Abstract

The invention belongs to the field of new materials, and particularly relates to a preparation method of a micro-nano multi-compartment polysaccharide colloid material. According to the method, a solvent replacement mechanism and microfluidic shear regulation are combined, precipitation, spontaneous aggregation and interface coating of polysaccharide polymers are sequentially achieved through a multi-stage reaction path, and multi-compartment particles which are stable in structure and clear in space partition are constructed. The used raw materials are water-soluble polysaccharides or fat-soluble polysaccharides, functional molecules can be accurately loaded in different areas of the particles, cross interference of components is avoided, and the particles have good environmental adaptability and functional modularization characteristics. The method does not need an emulsion template or a surfactant, the operation process is mild, simple and convenient, the particle structure is uniform, and the method is suitable for large-scale continuous preparation and multi-scene application.

Description

Technical Field

[0001] The present invention belongs to the field of new materials, and relates to multi-compartment polysaccharide colloidal materials and their preparation technologies. In particular, it is a preparation method of micro-nano multi-compartment polysaccharide colloidal materials based on a multi-stage continuous nanoprecipitation method and microfluidic technology, which is applicable to fields such as biocatalysis, drug delivery, and functional materials. Background Art

[0002] In the fields of drug delivery and biomaterial construction, in recent years, there has been extensive attention on how to achieve the co-encapsulation and directional release of different functional components through carriers with complex structures. Especially in aspects such as mimicking organelle structures, co-loading and precise delivery of multiple drugs, multi-compartment colloidal materials have shown great potential. However, most of the currently widely used methods rely on colloidal particles constructed from synthetic polymers or lipid materials, which have certain limitations in terms of material source and biocompatibility, and are difficult to meet the specific biological application requirements in terms of size regulation and structural stability.

[0003] To overcome these drawbacks, different preparation methods have been proposed in the prior art: For example, patent application CN201980067275.8 discloses a technical solution for constructing multi-compartment nanocapsules using a two-stage emulsion method. By constructing a double emulsion system of water-in-oil and then oil-in-water, the co-encapsulation of hydrophilic and lipophilic components is achieved. This method can prepare nanostructures with an outer diameter of less than 1 μm and has a certain interfacial stability. Although this technology performs well in constructing interfacial configurations, due to the high sensitivity of the emulsion structure to external perturbations, further optimization is still needed in terms of encapsulation efficiency and stability improvement. Patent application CN202110226751.5 provides a method for constructing multi-compartment microspheres based on microfluidic gas shearing technology. The structural scale prepared by this method mainly focuses on the millimeter level. Patent application CN202310103893.1 discloses a preparation method of calcium alginate multi-compartment hydrogel microcarriers based on biphasic droplet fusion. The raw materials used in this method have good biocompatibility and are suitable for constructing carrier structures from millimeters to centimeters. Patent application CN202310738189.3 proposes a method for constructing a multi-compartment ferritin delivery system using pH response and genipin cross-linking reaction. This method does not rely on templates or emulsions, but the construction materials are limited to animal proteins. Patent application CN201080031390.9 discloses a method for constructing multi-compartment nanoparticles based on multiple thermal cycles of functional amphiphiles. The material structure constructed by this method has good stability and is suitable for multi-drug combination delivery. However, when heat treatment is involved, the thermal sensitivity of bioactive substances needs to be considered, and at the same time, the safety of the metal complexes used in vivo needs to be comprehensively evaluated.

[0004] Although the existing preparation technologies of multi-compartment colloidal materials have made certain progress in functional loading and structure construction, there are generally problems such as poor structural stability, inaccurate size control, and poor compartment repeatability. In particular, it is difficult to achieve fine regulation and functional partitioning of compartments at the micro-nano scale. At the same time, the existing methods often rely on emulsion templates, cross-linking agents, or surfactants, resulting in complex system operations and easy cross-interference of components, making it difficult to meet the requirements of high stability and repeatability in biomedical scenarios. To overcome the above technical bottlenecks, the present invention proposes a continuous preparation method of multi-compartment colloidal materials based on the synergistic effect of microfluidics and solution replacement mechanism, introducing a three-stage reaction process, and gradually constructing a multi-compartment structure with clear boundaries through the sequential precipitation, aggregation, and coating of materials. This method can adopt a series or parallel microfluidic path according to requirements, realizing the orderly introduction and spatial positioning of functional components at different reaction stages, thereby improving the structural resolution and functional modularization level between compartments. Compared with traditional preparation methods, the method of the present invention does not require template or surfactant assistance, has strong system stability, and the construction process is mild and controllable, suitable for the precise assembly of polysaccharide colloids at the micro-nano scale. In addition, the formed multi-compartment structure can achieve the directional loading and coexistence of different components in each compartment according to functional requirements, significantly improving the practical performance of the material in complex application scenarios such as biological signal transmission, drug delivery, and multi-modal response, and having good scalability and industrial transformation potential. Summary of the Invention

[0005] The present invention aims to provide a continuous preparation method of micro-nano multi-compartment polysaccharide colloidal materials to solve the problems of poor structural stability, insufficient size uniformity, inaccurate compartment control, and easy cross-interference of functional components existing in the prior art. The method is based on a microfluidic control system, utilizes the principle of solution replacement between organic / water phases, and induces the interfacial response behavior of polymer materials through liquid-phase exchange under continuous flow conditions to construct multi-compartment colloidal particles. This method adopts a three-stage reaction path to gradually form a stable structure with spatial partitioning and functional separation. Compared with traditional emulsion template methods or physical encapsulation means, the present invention does not require the introduction of surfactants or particle templates, and the structure formation process relies on solvent conversion between liquid phases and interfacial tension regulation. The operation process is highly controllable, has good size adjustment ability, component adaptability, and the potential for large-scale continuous preparation. The technical solution adopted by the present invention is as follows: The present invention provides a method for preparing micro-nano (100 nm - 10 μm) multi-compartment polysaccharide colloidal particles based on a three-stage continuous nano-precipitation and microfluidic self-assembly strategy. The colloidal particles include an internal multi-compartment structure and a shell encapsulating the multi-compartment structure. The preparation process and features are as follows: The first, second, and third multi-channel micromixers are connected in series. In the first stage, the polysaccharide and the anti-solvent are rapidly mixed in the multi-channel micromixer to induce the nucleation of polysaccharide polymers and form primary nanoparticles. At the same time, the directional loading of functional substances in the compartments can be achieved by co-mixing functional molecules (such as proteins, active molecules, or drug molecules, etc.). In the second stage, the products are output in parallel through multiple multi-channel micromixers. In a salt-containing system, the electrostatic repulsion between particles is weakened by ion compression of the double electric layer, promoting the spontaneous aggregation of primary particles to construct heterogeneous particle aggregates. In the third stage, the aggregates are mixed with free polysaccharide again to trigger interfacial deposition, forming a coating structure to prevent the further growth of aggregates, thereby constructing a stable multi-compartment structure. Different functional components can be loaded within the same compartment, different compartments, or between multiple compartments of the multi-compartment colloidal particles. The method uses water-soluble polysaccharides and lipid-soluble polysaccharides to construct the multi-compartment structure. The method precisely regulates the phase separation behavior of the polymer through microfluidic technology, and adopts the solvent replacement method and shear force control to make the formed multi-compartment structure have good stability.

[0006] The preparation method of the water-soluble multi-compartment polysaccharide colloidal material involved in the present invention includes the following process: In the first stage, the water-soluble polysaccharide is added to water, stirred and heated to dissolve to obtain a polysaccharide aqueous solution. In the first-stage multi-channel micromixer, the water-soluble polysaccharide solution and the organic solvent are rapidly mixed through a multi-channel injection pump to trigger the nucleation and aggregation of water-soluble polysaccharide chains, and in-situ cross-linking is achieved through ion complexation to finally obtain water-soluble polysaccharide-based nanoparticles.

[0007] In the second stage, multiple first-stage multi-channel micromixers are connected in parallel, and their products are synchronously introduced into the second-stage multi-channel micromixer through different channels and rapidly mixed with the salt ion solution. The shielding effect of the salt ions induces the spontaneous aggregation of the nanoparticles, and aggregates of the nanoparticles are formed in the outlet flow of the second-stage multi-channel micromixer.

[0008] In the third stage, the second-stage multi-channel micromixer and the third-stage multi-channel micromixer are connected in series, and the particle aggregates are mixed with the polysaccharide aqueous solution and the organic reagent in the third-stage multi-channel micromixer to promote the supersaturated precipitation of free water-soluble polysaccharide chains at the interface of the aggregates, thereby further stabilizing the compartment structure. The organic solvent is removed by dialysis to obtain stable glycosyl multi-compartment polysaccharide colloidal particles.

[0009] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: In the first stage, an aqueous polysaccharide solution (channel 1), an aqueous solution with cross - linking function (channel 3), and organic reagents (channels 2 and 4) are rapidly mixed in a multi - channel micro - mixer at the first stage using a multi - channel syringe pump to obtain an aqueous solution of polysaccharide - based nanoparticles in the outlet flow of the multi - channel micro - mixer at the first stage.

[0010] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: In the second stage, multiple multi - channel micro - mixers at the first stage are installed in parallel, and their outflow products are simultaneously fed into a multi - channel micro - mixer at the second stage through different inlets (channels 1, 2, 3) and rapidly mixed with a solution containing salt ions (channel 4). The shielding effect of salt ions induces the spontaneous aggregation of nanoparticles, and an aqueous solution of polysaccharide - based nanoparticle aggregates is obtained in the outlet flow of the multi - channel micro - mixer at the second stage.

[0011] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: In the third stage, the multi - channel micro - mixer at the second stage and the multi - channel micro - mixer at the third stage are connected in series. Nanoparticle aggregates (channels 1 and 3), an aqueous polysaccharide solution (channel 2), and organic reagents (channel 4) are finally mixed in the multi - channel micro - mixer at the third stage, triggering the supersaturation of free water - soluble polysaccharide chains and their precipitation at the aggregate interface.

[0012] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: The flow rate is kept constant at 5 - 40 mL / min in all three stages of preparation.

[0013] The preparation method of the fat - soluble multi - compartment polysaccharide colloidal material involved in the present invention includes the following processes: In the first stage, fat - soluble polysaccharide is added to an organic solvent to dissolve and obtain a polysaccharide solution. In a multi - channel micro - mixer at the first stage, the fat - soluble polysaccharide solution is rapidly mixed with an aqueous phase through a multi - channel syringe pump. The formation of fat - soluble polysaccharide - based nanoparticles is promoted by the solvent replacement and self - assembly mechanisms, and the particle structure is stabilized by hydrogen - bonding and hydrophobic interactions to obtain a uniformly dispersed fat - soluble polysaccharide - based nanoparticles.

[0014] In the second stage, multiple multi - channel micro - mixers at the first stage are connected in parallel, and their products are synchronously fed into a multi - channel micro - mixer at the second stage through different channels and rapidly mixed with a salt - ion solution. The shielding effect of salt ions induces the spontaneous aggregation of nanoparticles, and aggregates of nanoparticles are formed in the outlet flow of the multi - channel micro - mixer at the second stage.

[0015] In the third stage, the multi - channel micro - mixer at the second stage and the multi - channel micro - mixer at the third stage are connected in series, so that the particle aggregates are mixed with the fat - soluble polysaccharide solution and the aqueous phase in the multi - channel micro - mixer at the third stage, promoting the supersaturation precipitation of free fat - soluble polysaccharide chains at the aggregate interface, thereby further stabilizing the compartment structure. The organic solvent and salt ions are removed by dialysis to obtain stable fat - soluble polysaccharide - based multi - compartment polysaccharide colloidal particles.

[0016] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: In the first stage, the fat - soluble polysaccharide organic solution (channel 1), organic solvent (channel 3), and water (channels 2 and 4) are rapidly mixed in a multi - channel micro - mixer at the first stage using a multi - channel syringe pump to obtain fat - soluble polysaccharide - based nanoparticles in the outlet flow of the multi - channel micro - mixer at the first stage.

[0017] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: In the second stage, multiple multi - channel micro - mixers at the first stage are installed in parallel. The outflow products are simultaneously merged into a multi - channel micro - mixer at the second stage through different inlets (channels 1, 2, 3) and rapidly mixed with a solution containing salt ions (channel 4). The shielding effect of the salt ions induces the spontaneous aggregation of the nanoparticles, and aggregates of fat - soluble polysaccharide - based nanoparticles are obtained in the outlet flow of the multi - channel micro - mixer at the second stage.

[0018] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: In the third stage, the multi - channel micro - mixer at the second stage and the multi - channel micro - mixer at the third stage are connected in series. The nanoparticle aggregates (channels 1 and 3), fat - soluble polysaccharide organic solution (channel 2), and water (channel 4) are mixed in the multi - channel micro - mixer at the third stage, triggering the supersaturation of free fat - soluble polysaccharide chains and precipitation at the interface of the aggregates.

[0019] Based on the above - mentioned solution and as a preferred solution of the above - mentioned solution: In all three stages of the preparation, the flow rate is maintained at 5 - 40 mL / min.

[0020] The beneficial effects of the present invention are as follows: 1. The raw materials used in the present invention are composed of water - soluble polysaccharides or fat - soluble polysaccharides, which have good hydrophilicity and hydrophobicity respectively, and can achieve the precise distribution and efficient loading of multiple functional components in a multi - compartment structure. When applied to complex environments, different components can be reasonably separated in space to avoid cross - interference.

[0021] 2. The process method of the present invention is simple and efficient. By using three - stage continuous nanoprecipitation combined with microfluidic self - assembly technology, the construction of multi - compartment particles is realized under mild conditions. Through ion - induced aggregation and solvent replacement, a dense shell structure is formed, enabling the particles to have good structural integrity and molding stability. Compared with traditional batch methods, this method has higher controllability and repeatability, and the structure can be customized by adjusting microfluidic parameters.

[0022] 3. The multi - compartment colloidal particles constructed by the present invention have high stability in structural design. The shell is formed by polysaccharide - induced interfacial precipitation, combined with the shielding effect of salt ions, which can significantly improve the integrity of the particles in different environments. When placed in an environment with a pH of 3 - 11 for a long time, the structure shows no obvious collapse or rupture, meeting the usage requirements in complex application scenarios.

[0023] 4. The multi-compartment structure constructed by the present invention has a high degree of spatial controllability, and can position different components in the same compartment, different compartments, or between multiple compartments according to the required functional requirements. The method of the present invention realizes the organic integration of the loading path of functional components and the spatial structure during the structure construction process, providing stronger structural adaptability and functional flexibility for subsequent applications such as signal response, biological delivery, or multi-functional material integration. Description of the Drawings

[0024] Figure 1 is a multi-channel micromixer; Figure 2 is a schematic flow chart of the preparation method of the micro-nano multi-compartment polysaccharide colloidal particles involved in the present invention; Figure 3 is a transmission electron microscope characterization diagram of the structure of the prepared acetylated dextran-based multi-compartment polysaccharide colloidal particles; Figure 4 is a transmission electron microscope characterization diagram of the structure of the prepared sodium carboxymethyl cellulose-based multi-compartment polysaccharide colloidal particles. Detailed Embodiments

[0025] The technical solution of the present invention will be further described in conjunction with the accompanying drawings and specific embodiments. In the description of the solution, some exemplary embodiments of the present invention are described by way of illustration. Those of ordinary skill in the art can recognize that the described embodiments can be modified in various different ways without departing from the claims of the present invention.

[0026] At the same time, similar words such as "including" and "comprising" in the specification should be interpreted as having an inclusive meaning rather than an exclusive or exhaustive meaning; that is, it means including but not limited to. Therefore, the drawings and the description are illustrative in nature and are not used to limit the protection scope of the claims.

[0027] The present invention proposes to precisely construct multi-compartment polysaccharide colloidal particles based on a three-stage continuous nanoprecipitation process, and realize the directional loading of functional components at different stages. The preparation process of the colloidal particles proposed by the present invention is as Figure 2 shown. The first stage is in a multi-channel micromixer ( Figure 1)Rapid mixing is carried out to complete the preliminary nucleation of nanoparticles and the encapsulation of payloads such as proteins, active molecules or drug molecules, etc., to achieve loading inside the compartments; the second stage induces the aggregation of heterogeneous particles through a multi-channel parallel device to construct a multi-functional compartment system; the third stage further introduces functional materials on the surface of the aggregates or between the membranes to achieve loading between compartments. By adjusting the types of raw materials and channel configurations, the precise distribution of different functional components alone or in combination in different compartments can be regulated, so as to meet the requirements of complex functions such as cascade reactions, signal amplification and energy conduction inside the material particles and between particles. This method demonstrates the advantages of staged controllable loading in the construction of multi-functional nano-systems.

[0028] Unless otherwise specified, the experimental methods described in the examples are all conventional methods; the reagents and materials, unless otherwise specified, can be obtained from commercial channels.

[0029] Example 1: Preparation of acetylated dextran-based multi-compartment polysaccharide colloidal particles In the first stage, an acetone solution of acetylated dextran (1 mg / mL, channel 1), acetone (channel 3), and water (channels 2 and 4) are rapidly mixed in a one-stage multi-channel micromixer using a multi-channel syringe pump to prepare acetylated dextran-based nanoparticles. In the second stage, two one-stage multi-channel micromixers are installed in parallel, and the effluents are simultaneously introduced into a two-stage multi-channel micromixer through different inlets (channels 1 and 3), and rapidly mixed with a 600 mmol / L aqueous sodium chloride solution (channel 2) and acetone (channel 4) to form aggregates of nanoparticles in the outlet stream of the two-stage multi-channel micromixer. In the third stage, the two-stage and three-stage multi-channel micromixers are connected in series. The nanoparticle aggregates (channels 1 and 3), an acetylated dextran solution (1 mg / mL, channel 2), and water (channel 4) are mixed in the three-stage multi-channel micromixer to obtain acetylated dextran-based multi-compartment polysaccharide colloidal particles. After removing acetone and sodium chloride by dialysis, the particle size and dispersibility of the particles are detected by dynamic light scattering analysis; the morphology of the particles is characterized by transmission electron microscopy. The large particle size of the acetylated dextran-based nanoparticles characterized by dynamic light scattering is 120 nm, and the dispersibility is 0.11. The particle size of the multi-compartment polysaccharide colloidal particles is 813 nm, and the dispersibility is 0.21. In the transmission electron microscopy image, the acetylated dextran-based multi-compartment polysaccharide colloidal particles exhibit a uniform spherical structure. Due to the collapse of the polymer shell during the dehydration process, the inner core acetylated dextran nanoparticles of the acetylated dextran-based multi-compartment polysaccharide colloidal particles can be clearly seen ( Figure 3 ). There is a certain degree of aggregation between the particles, and obvious multi-compartment characteristics can be observed on the surface and inside of the particles. These compartment structures present stable interfaces and have good morphological integrity.

[0030] Example 2: Preparation of sodium carboxymethyl cellulose-based multi-compartment polysaccharide colloidal particles At the first stage, an aqueous solution of sodium carboxymethylcellulose (1 mg / mL, channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), and ethanol (channels 2 and 4) were rapidly mixed in a one-stage multi-channel micromixer using a multi-channel syringe pump to prepare sodium carboxymethylcellulose-based nanoparticles. At the second stage, two one-stage multi-channel micromixers were installed in parallel, and the effluents were simultaneously introduced into the second-stage multi-channel micromixer through different inlets (channels 1 and 3), and rapidly mixed with an aqueous solution of 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the outlet stream of the second-stage multi-channel micromixer. At the third stage, the second-stage and third-stage multi-channel micromixers were connected in series. The nanoparticle aggregates (channels 1 and 3), the sodium carboxymethylcellulose solution (1 mg / mL, channel 2), and ethanol (channel 4) were finally mixed in the third-stage multi-channel micromixer to obtain sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersity were detected by dynamic light scattering analysis; the morphology of the particles was characterized by transmission electron microscopy. The particle size of the sodium carboxymethylcellulose-based nanoparticles was 365 nm and the dispersity was 0.21, and the particle size of the multi-compartment polysaccharide colloidal particles was 987 nm and the dispersity was 0.33 as characterized by dynamic light scattering. In the transmission electron micrograph, the sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles exhibited a uniform spherical structure ( Figure 4 )

[0031] Example 3: Loading of the Lipophilic Compound Nile Red into Sodium Carboxymethylcellulose-Based Multi-Compartment Polysaccharide Colloidal Particles At the first stage, an aqueous solution of sodium carboxymethylcellulose (1 mg / mL, channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), a Nile red ethanol solution (channel 2), and ethanol (channel 4) were rapidly mixed in a one-stage multi-channel micromixer using a multi-channel syringe pump to prepare sodium carboxymethylcellulose-based nanoparticles. At the second stage, two one-stage multi-channel micromixers were installed in parallel, and the effluents were simultaneously introduced into the second-stage multi-channel micromixer through different inlets (channels 1 and 3), and rapidly mixed with an aqueous solution of 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the outlet stream of the second-stage multi-channel micromixer. At the third stage, the second-stage and third-stage multi-channel micromixers were connected in series. The nanoparticle aggregates (channels 1 and 3), the sodium carboxymethylcellulose solution (1 mg / mL, channel 2), and ethanol (channel 4) were finally mixed in the third-stage multi-channel micromixer to obtain sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersity were detected by dynamic light scattering analysis; the morphology of the particles was characterized by transmission electron microscopy.

[0032] Example 4: Acetylated dextran-based multi-compartment polysaccharide colloidal particles loaded with lipophilic compound Nile red In the first stage, an acetone solution of acetylated dextran (1 mg / mL, channel 1) and an acetone solution of Nile red (channel 3), and water (channels 2 and 4) were rapidly mixed in a multi-channel micro-mixer at the first stage using a multi-channel syringe pump to prepare acetylated dextran-based nanoparticles. In the second stage, two multi-channel micro-mixers at the first stage were installed in parallel, and the effluents were simultaneously introduced into the multi-channel micro-mixer at the second stage through different inlets (channels 1 and 3), and rapidly mixed with an aqueous solution of 600 mmol / L sodium chloride (channel 2) and acetone (channel 4) to form aggregates of nanoparticles in the outlet stream of the multi-channel micro-mixer at the second stage. In the third stage, the multi-channel micro-mixer at the second stage and the multi-channel micro-mixer at the third stage were connected in series. The nanoparticle aggregates (channels 1 and 3), an acetylated dextran solution (1 mg / mL, channel 2), and water (channel 4) were mixed in the multi-channel micro-mixer at the third stage to obtain acetylated dextran-based multi-compartment polysaccharide colloidal particles. After removing acetone and sodium chloride by dialysis, the particle size and dispersity of the particles were detected by dynamic light scattering analysis; the morphology of the particles was characterized by transmission electron microscopy.

[0033] Example 5: Sodium carboxymethyl cellulose-based multi-compartment polysaccharide colloidal particles loaded with protein glucose oxidase In the first stage, an aqueous solution of sodium carboxymethyl cellulose (1 mg / mL) and glucose oxidase (channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), ethanol (channel 2), and ethanol (channel 4) were rapidly mixed in a multi-channel micro-mixer at the first stage using a multi-channel syringe pump to prepare sodium carboxymethyl cellulose-based nanoparticles. In the second stage, two multi-channel micro-mixers at the first stage were installed in parallel, and the effluents were simultaneously introduced into the multi-channel micro-mixer at the second stage through different inlets (channels 1 and 3), and rapidly mixed with an aqueous solution containing 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the outlet stream of the multi-channel micro-mixer at the second stage. In the third stage, the multi-channel micro-mixer at the second stage and the multi-channel micro-mixer at the third stage were connected in series. The nanoparticle aggregates (channels 1 and 3), a sodium carboxymethyl cellulose solution (1 mg / mL, channel 2), and ethanol (channel 4) were mixed in the multi-channel micro-mixer at the third stage to obtain sodium carboxymethyl cellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersity of the particles were detected by dynamic light scattering analysis; the morphology of the particles was characterized by transmission electron microscopy.

[0034] Example 6: Sodium carboxymethyl cellulose-based multi-compartment polysaccharide colloidal particles loaded with protein catalase In the first stage, sodium carboxymethylcellulose (1 mg / mL) and an aqueous solution of catalase (channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), ethanol (channel 2), and ethanol (channel 4) were rapidly mixed in a multi-channel micro-mixer at the first stage using a multi-channel syringe pump to prepare sodium carboxymethylcellulose-based nanoparticles. In the second stage, two multi-channel micro-mixers at the first stage were installed in parallel, and the effluents were simultaneously introduced into the multi-channel micro-mixer at the second stage through different inlets (channels 1 and 3), and rapidly mixed with an aqueous solution of 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the outlet stream of the multi-channel micro-mixer at the second stage. In the third stage, the multi-channel micro-mixer at the second stage was connected in series with the multi-channel micro-mixer at the third stage. The nanoparticle aggregates (channels 1 and 3), a sodium carboxymethylcellulose solution (1 mg / mL, channel 2), and ethanol (channel 4) were mixed in the multi-channel micro-mixer at the third stage to obtain sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersibility of the particles were detected by dynamic light scattering analysis; the morphology of the particles was characterized using a transmission electron microscope.

[0035] Example 7: Sodium Carboxymethylcellulose-Based Multi-Compartment Polysaccharide Colloidal Particles Loaded with Protein Horseradish Peroxidase and Peroxidase Fluorescent Probe In the first stage, sodium carboxymethylcellulose (1 mg / mL) and an aqueous solution of horseradish peroxidase (channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), an ethanol solution of peroxidase fluorescent probe (channel 2), and ethanol (channel 4) were rapidly mixed in a multi-channel micro-mixer at the first stage using a multi-channel syringe pump to prepare sodium carboxymethylcellulose-based nanoparticles. In the second stage, two multi-channel micro-mixers at the first stage were installed in parallel, and the effluents were simultaneously introduced into the multi-channel micro-mixer at the second stage through different inlets (channels 1 and 3), and rapidly mixed with an aqueous solution of 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the outlet stream of the multi-channel micro-mixer at the second stage. In the third stage, the multi-channel micro-mixer at the second stage was connected in series with the multi-channel micro-mixer at the third stage. The nanoparticle aggregates (channels 1 and 3), a sodium carboxymethylcellulose solution (1 mg / mL, channel 2), and ethanol (channel 4) were mixed in the multi-channel micro-mixer at the third stage to obtain sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersibility of the product were detected by dynamic light scattering analysis.

[0036] Example 8: Sodium Carboxymethylcellulose-Based Multi-Compartment Polysaccharide Colloidal Particles Loaded with Glucose Oxidase (inside the Compartment) and Catalase (between the Compartments) In the first stage, sodium carboxymethylcellulose (1 mg / mL) and an aqueous solution of glucose oxidase (channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), ethanol (channel 2), and ethanol (channel 4) were rapidly mixed in a first-stage multi-channel micromixer using a multi-channel syringe pump to prepare sodium carboxymethylcellulose-based nanoparticles. In the second stage, two first-stage multi-channel micromixers were installed in parallel, and their effluent products were simultaneously introduced into a second-stage multi-channel micromixer through different inlets (channels 1 and 3) and rapidly mixed with an aqueous solution containing 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the outlet stream of the second-stage multi-channel micromixer. In the third stage, the second-stage and third-stage multi-channel micromixers were connected in series. The nanoparticle aggregates (channels 1 and 3), sodium carboxymethylcellulose (1 mg / mL), and an aqueous solution of catalase (channel 2) and ethanol (channel 4) were mixed in the third-stage multi-channel micromixer to obtain sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersity of the particles were detected by dynamic light scattering analysis, and the morphology of the particles was characterized using a transmission electron microscope.

[0037] Example 9: Sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles loaded with glucose oxidase (inside the compartment), horseradish peroxidase / peroxidase fluorescent probe (in the adjacent compartment), and catalase (between the compartments) In the first stage, sodium carboxymethylcellulose (1 mg / mL) and an aqueous solution of glucose oxidase (channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), ethanol (channel 2), and ethanol (channel 4) were rapidly mixed using a multi-channel syringe pump in a first-stage multi-channel micromixer to prepare sodium carboxymethylcellulose-based nanoparticles loaded with glucose oxidase. Sodium carboxymethylcellulose (1 mg / mL) and an aqueous solution of horseradish peroxidase (channel 1), an aqueous solution of calcium chloride (0.05 mg / mL, channel 3), an ethanol solution of a peroxidase fluorescent probe (channel 2), and ethanol (channel 4) were rapidly mixed using a multi-channel syringe pump in a first-stage multi-channel micromixer to prepare sodium carboxymethylcellulose-based nanoparticles loaded with horseradish peroxidase / peroxidase fluorescent probe. In the second stage, two first-stage multi-channel micromixers were installed in parallel, and the effluents passed through different inlets. The sodium carboxymethylcellulose-based nanoparticles loaded with glucose oxidase (channel 1) and the sodium carboxymethylcellulose-based nanoparticles loaded with horseradish peroxidase / peroxidase fluorescent probe (channel 3) were simultaneously introduced into a second-stage multi-channel micromixer and rapidly mixed with an aqueous solution of 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the effluent stream of the second-stage multi-channel micromixer. In the third stage, the second-stage and third-stage multi-channel micromixers were connected in series. The nanoparticle aggregates (channels 1 and 3), sodium carboxymethylcellulose (1 mg / mL), and an aqueous solution of catalase (channel 2) and ethanol (channel 4) were mixed in a third-stage multi-channel micromixer to obtain sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersibility of the particles were detected using dynamic light scattering analysis.

[0038] Example 10: Sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles loaded with glucose oxidase (inside the compartment) and horseradish peroxidase / peroxidase fluorescent probe (in the adjacent compartment) At the first stage, sodium carboxymethylcellulose (1 mg / mL) and an aqueous glucose oxidase solution (channel 1), an aqueous calcium chloride solution (0.05 mg / mL, channel 3), ethanol (channel 2), and ethanol (channel 4) were rapidly mixed in a first-stage multi-channel micromixer using a multi-channel syringe pump to prepare sodium carboxymethylcellulose-based nanoparticles loaded with glucose oxidase. Sodium carboxymethylcellulose (1 mg / mL) and an aqueous horseradish peroxidase solution (channel 1), an aqueous calcium chloride solution (0.05 mg / mL, channel 3), an ethanol solution of a catalase fluorescent probe (channel 2), and ethanol (channel 4) were rapidly mixed in a first-stage multi-channel micromixer using a multi-channel syringe pump to prepare sodium carboxymethylcellulose-based nanoparticles loaded with horseradish peroxidase / catalase fluorescent probe. At the second stage, two first-stage multi-channel micromixers were installed in parallel, and the effluents passed through different inlets. The sodium carboxymethylcellulose-based nanoparticles loaded with glucose oxidase (channel 1) and the sodium carboxymethylcellulose-based nanoparticles loaded with horseradish peroxidase / catalase fluorescent probe (channel 3) were simultaneously introduced into a second-stage multi-channel micromixer and rapidly mixed with an aqueous solution containing 600 mmol / mL sodium chloride and 0.1 mg / mL calcium chloride (channel 2) and ethanol (channel 4) to form aggregates of nanoparticles in the outlet stream of the second-stage multi-channel micromixer. At the third stage, the second-stage and third-stage multi-channel micromixers were connected in series. The nanoparticle aggregates (channels 1 and 3), a sodium carboxymethylcellulose solution (1 mg / mL, channel 2), and ethanol (channel 4) were mixed in a third-stage multi-channel micromixer to obtain sodium carboxymethylcellulose-based multi-compartment polysaccharide colloidal particles. After removing ethanol and sodium chloride by dialysis, the particle size and dispersity of the particles were detected using dynamic light scattering analysis.

[0039] Some specific embodiments of the present invention have been described above. It should be understood that those of ordinary skill in the art can make many modifications to the method of the present invention without creative work. Therefore, all technical solutions that can be obtained by those skilled in the art in the technical field of the present invention based on the method of the present invention through logical analysis, reasoning, or limited experiments on the basis of the prior art are within the protection scope determined by the claims.

Claims

1. A method for preparing a micro-nano multi-compartment polysaccharide colloid material, characterized in that: Stable micro-nano multi-compartment structured particles are prepared using multi-stage continuous nanoprecipitation and microfluidic self-assembly technology.

2. The method for preparing multi-compartment polysaccharide colloidal particles according to claim 1, characterized in that: The multi-stage multi-channel micromixers are connected in series.

3. The method for preparing multi-compartment polysaccharide colloidal particles according to claim 1, characterized in that: In the first stage, the polysaccharide solution and the antisolvent phase are rapidly mixed in a multi-channel micromixer to induce the nucleation of polysaccharide polymers and form primary nanoparticles.

4. The method for preparing multi-compartment polysaccharide colloidal particles according to claim 1, characterized in that: The second stage outputs the product through multiple multi-channel micromixers in parallel. In the salt-containing system, the ions compress the double electric layer to weaken the electrostatic repulsion between particles, promote spontaneous aggregation between primary particles, and construct nanoparticle aggregates.

5. The method for preparing multi-compartment polysaccharide colloidal particles according to claim 1, characterized in that: In the third stage, the aggregates are mixed with free polysaccharides again to induce interfacial deposition and form a membrane structure, which prevents the further growth of the aggregates and thus constructs a stable multi-compartment structure.

6. The method for preparing multi-compartment polysaccharide colloidal particles according to claim 1, characterized in that: The first, second and third stages can achieve directional loading of functional substances within the compartments by blending loads, such as proteins, active molecules or drug molecules.

7. A method for preparing micro-nano multi-compartment polysaccharide colloidal particles, characterized in that: Different components can be loaded in the same compartment, in different compartments, or between multiple compartments of the multi-compartment colloidal particles.

8. A method for preparing micro-nano multi-compartment polysaccharide colloidal particles, characterized in that: The method utilizes water-soluble polysaccharides or fat-soluble polysaccharides to construct a multi-compartment structure.

9. A method for preparing micro-nano multi-compartment polysaccharide colloidal particles, characterized in that: Microfluidic technology is combined with the solvent replacement process to precisely control the phase separation behavior of the polymer and construct a structurally stable multi-compartment structure.

10. The method for preparing multi-compartment polysaccharide colloidal particles according to claim 9, characterized in that: The size of the prepared multi-compartment structure can be controlled by regulating the microfluidic parameters.

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