Multi-channel micro-fluidic chip and method for preparing curcumin-loaded nanoparticles by using multi-channel micro-fluidic chip

Abstract

The invention discloses a multi-channel micro-fluidic chip which comprises a chip body, at least two layers of flow channel networks are arranged in the chip body from top to bottom, the two layers of flow channel networks are both of a binary tree structure, a tip flow channel of the lower layer of flow channel network is communicated with a reaction chamber, a tip flow channel of the upper layer of flow channel network is communicated with the reaction chamber through a reduced-order flow channel, and the lower layer of flow channel network is communicated with the reaction chamber through a second-order flow channel. The fluid in the upper-layer flow channel network and the fluid in the lower-layer flow channel network are mixed and reacted in the reaction chamber; wherein a root flow channel of the upper-layer flow channel network is communicated with a first liquid inlet flow channel penetrating to the surface of the chip body, a root flow channel of the lower-layer flow channel network is communicated with a second liquid inlet flow channel penetrating to the surface of the chip body, and the reaction chamber is communicated with a liquid outlet flow channel penetrating to the surface of the chip body. The invention also discloses a method for preparing curcumin-loaded nanoparticles by using the multi-channel micro-fluidic chip. The curcumin-loaded nanoparticles with good size and form uniformity and high load rate can be prepared.

Description

Technical Field

[0001] The present invention relates to the technical field of nanoparticle preparation, and more specifically, to a multi-channel microfluidic chip and a method for preparing curcumin-loaded nanoparticles using the same. Background Art

[0002] In recent years, microfluidic chip technology has been widely used in biomedicine, chemical analysis, drug development and other fields because it can achieve micro-quantification of samples, high-throughput screening and precise control of reaction conditions. In particular, multi-channel microfluidic chips can process multiple samples or reactions at the same time, greatly improving experimental efficiency and accuracy.

[0003] Nanoparticles, as a new type of drug delivery carrier, have attracted attention because they can improve the solubility, bioavailability and targeting of drugs. However, traditional methods for preparing nanoparticles in batches often have problems such as uneven particle size distribution, complex preparation process, and difficulty in large-scale production. These challenges limit the potential of nanoparticles in clinical applications.

[0004] Curcumin, a natural compound extracted from turmeric rhizomes, has a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer effects. However, its clinical application is limited due to its low water solubility and poor bioavailability. To overcome these problems, it becomes crucial to develop an efficient and stable curcumin delivery system.

[0005] Zein, as a natural biopolymer, has become an ideal material for preparing drug delivery systems due to its good biocompatibility, biodegradability and controllable physicochemical properties. It has been proven that it can effectively load and stabilize curcumin and improve its solubility.

[0006] Although studies have reported methods for preparing curcumin nanoparticles using zein, most technologies still face some limitations, such as the difficulty in precisely controlling the size and morphology of nanoparticles and the lack of efficient and precise preparation platforms. In addition, traditional preparation methods are often time-consuming and difficult to automate and achieve high-throughput production. Summary of the invention

[0007] The purpose of the present invention is to provide a multi-channel microfluidic chip to provide a platform for producing curcumin-loaded nanoparticles with relatively uniform size and morphology.

[0008] Another object of the present invention is to provide a method for preparing curcumin-loaded nanoparticles using the multi-channel microfluidic chip, so as to prepare curcumin-loaded nanoparticles with relatively uniform size and morphology.

[0009] In order to achieve these purposes and other advantages according to the present invention, a multi-channel microfluidic chip is provided, comprising:

[0010] The chip body has at least two layers of flow channel networks arranged from top to bottom, both of which are in a binary tree structure, the terminal flow channels of the lower flow channel network are connected to the reaction chamber, and the terminal flow channels of the upper flow channel network are connected to the reaction chamber through a descending flow channel, so as to make the fluid in the upper flow channel network and the fluid in the lower flow channel network mix and react in the reaction chamber;

[0011] The root channel of the upper channel network is connected to a first liquid inlet channel that penetrates the surface of the chip body, the root channel of the lower channel network is connected to a second liquid inlet channel that penetrates the surface of the chip body, and the reaction chamber is connected to a liquid outlet channel that penetrates the surface of the chip body;

[0012] The width of the distal flow channels of the upper layer flow channel network is 150 to 250 μm, and the width of the distal flow channels of the lower layer flow channel network is 290 to 500 μm.

[0013] Preferably, the chip body includes a first sheet, a second sheet and a third sheet stacked from top to bottom, the upper channel network is arranged between the first sheet and the second sheet, and the lower channel network, the reaction chamber and the liquid outlet channel are arranged between the second sheet and the third sheet.

[0014] Preferably, the terminal flow channel of the upper flow channel network is provided with 2 n The terminal flow channel of the lower flow channel network is provided with 2 n+1 , n>0, the reaction chamber is provided with 2 n Each of the two distal flow channels of the lower layer flow channel network is connected to the same reaction chamber as a group, and the step-down flow channel is provided with 2 n and correspond to the 2 of the upper flow channel network respectively. n The terminal flow channels correspond to 2 n The step-down flow channel is L-shaped, the vertical portion of which is connected to the corresponding terminal flow channel of the upper flow channel network, and the horizontal portion is sandwiched between a group of terminal flow channels of the lower flow channel network and connected to the corresponding reaction chamber. The liquid outlet flow channel includes two corresponding channels connected to each reaction chamber. n The outlet liquid flow channel and 2 n The outlet of the total liquid outlet flow channel is arranged at the end surface of the chip body.

[0015] Preferably, the first sheet material, the second sheet material and the third sheet material are all made of glass.

[0016] The present invention also provides a method for preparing curcumin-loaded nanoparticles using the multi-channel microfluidic chip, comprising the following steps:

[0017] Step 1, dissolving zein and curcumin in ethanol, and dispersing by ultrasonication to obtain a mixed alcohol solution of zein and curcumin;

[0018] Step 2: using the mixed alcohol solution as the inner phase and the ultrapure aqueous solution as the outer phase, passing the inner phase into the first liquid inlet channel of the multi-channel microfluidic chip, passing the outer phase into the second liquid inlet channel of the multi-channel microfluidic chip, and obtaining a dispersion of zein nanoparticles loaded with curcumin from the liquid outlet channel of the multi-channel microfluidic chip;

[0019] Step 3: placing the zein nanoparticle dispersion loaded with curcumin in a dialysis bag for dialysis for a certain period of time, and freeze-drying to obtain the curcumin nanoparticles loaded with curcumin.

[0020] Preferably, in step 1, the concentration of zein is 5-40 mg / mL, the mass ratio of curcumin to zein is 1:5-50, and the volume fraction of ethanol is 60-95%.

[0021] Preferably, in step 2, the flow rate of the inner phase is 4 to 24 mL / h, and the flow rate of the outer phase is 80 to 300 mL / h.

[0022] Preferably, a 10KD dialysis bag is selected and the dialysis time is 24 to 72 hours.

[0023] The present invention also provides curcumin-loaded nanoparticles prepared by the method.

[0024] The present invention also provides the application of the above-mentioned curcumin-loaded nanoparticles in loading and delivering in food, medicine and health products.

[0025] The present invention at least has the following beneficial effects: the multi-channel microfluidic chip provided by the present invention provides a platform for realizing automated and high-throughput production of loaded curcumin nanoparticles with relatively uniform size and morphology and high loading rate. At the same time, the method for preparing loaded curcumin nanoparticles provided by the present invention can prepare loaded curcumin nanoparticles with good uniformity in size and morphology and high loading rate.

[0026] Other advantages, objectives and features of the present invention will be embodied in part through the following description, and in part will be understood by those skilled in the art through study and practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Schematic diagram of the structure of the multi-channel microfluidic chip according to an embodiment of the present invention;

[0028] Figure 2This is a particle size distribution diagram of the curcumin-loaded nanoparticles prepared in Example 1 of the present invention;

[0029] Figure 3 This is a particle size distribution diagram of the curcumin-loaded nanoparticles prepared in Example 2 of the present invention;

[0030] Figure 4 This is a particle size distribution diagram of the curcumin-loaded nanoparticles prepared in Example 3 of the present invention;

[0031] Figure 5 This is a particle size distribution diagram of the product obtained in Comparative Example 1 of the present invention;

[0032] Figure 6 This is a particle size distribution diagram of the product obtained in Comparative Example 2 of the present invention;

[0033] Figure 7 This is a particle size distribution diagram of the product obtained in Comparative Example 3 of the present invention;

[0034] Figure 8 This is a particle size distribution diagram of the product obtained in Comparative Example 4 of the present invention;

[0035] Fig. 9 This is the particle size distribution diagram of the product obtained in Comparative Example 5 of the present invention. DETAILED DESCRIPTION

[0036] The present invention is further described in detail below in conjunction with the accompanying drawings so that those skilled in the art can implement the invention with reference to the description.

[0037] It should be noted that the experimental methods described in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials are commercially available unless otherwise specified; in the description of the present invention, the terms "lateral", "longitudinal", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.

[0038] The present invention provides a multi-channel microfluidic chip, comprising:

[0039] The chip body has at least two layers of flow channel networks arranged from top to bottom, both of which are in a binary tree structure, the terminal flow channels of the lower flow channel network are connected to the reaction chamber, and the terminal flow channels of the upper flow channel network are connected to the reaction chamber through a descending flow channel, so as to make the fluid in the upper flow channel network and the fluid in the lower flow channel network mix and react in the reaction chamber;

[0040] Since the fluid is divided through the flow channel network, the components in the fluid are not easy to agglomerate, which is conducive to the formation of nanoparticles. In addition, the fluid is divided into the terminal flow channels parallel to each other in the flow channel network, and then mixed in the reaction chambers connected by the terminal flow channels, so that the amount of fluid in each reaction chamber is small, and sufficient diffusion and mixing can be carried out, thereby improving the loading rate of nanoparticles.

[0041] The root channel of the upper channel network is connected to a first liquid inlet channel that penetrates the surface of the chip body, the root channel of the lower channel network is connected to a second liquid inlet channel that penetrates the surface of the chip body, and the reaction chamber is connected to a liquid outlet channel that penetrates the surface of the chip body;

[0042] Here, the first liquid inlet flow channel can be used to add the inner phase fluid for preparing the loaded curcumin nanoparticles, the second liquid inlet flow channel can be used to add the outer phase fluid for preparing the loaded curcumin nanoparticles, and the liquid outlet flow channel is used to discharge the prepared loaded curcumin nanoparticles.

[0043] The width of the terminal flow channels of the upper layer flow channel network is 150 to 250 μm. At this width, the terminal flow channels of the upper layer flow channel network can effectively control the flow rate of the inner phase fluid to avoid excessive flow rate leading to excessive number of solvent molecules in the inner phase fluid and agglomeration; the width of the terminal flow channels of the lower layer flow channel network is 290 to 500 μm. At this width, the terminal flow channels of the lower layer flow channel network can just match the flow rate of the external phase fluid with the inner phase fluid to achieve a better dispersion effect.

[0044] Specifically, the chip body includes a first sheet, a second sheet and a third sheet stacked from top to bottom, the upper channel network is arranged between the first sheet and the second sheet, and the lower channel network, the reaction chamber and the liquid outlet channel are arranged between the second sheet and the third sheet.

[0045] When processing the chip body, grooves can be etched on the lower surface of the first sheet and the upper surface of the second sheet to form an upper layer flow channel network, and grooves can be etched on the lower surface of the second sheet and the upper surface of the third sheet to form a lower layer flow channel network. Since the flow channel network is not made by drilling holes in the chip body, but by etching grooves on the surface of the sheet, it is easier to make and it is also easier to control the accuracy of the flow channel width.

[0046] Specifically, the terminal flow channel of the upper flow channel network is provided with 2 n The terminal flow channel of the lower flow channel network is provided with 2 n+1 , n>0, the reaction chamber is provided with 2 n Each of the two distal flow channels of the lower layer flow channel network is connected to the same reaction chamber as a group, and the step-down flow channel is provided with 2 nand correspond to the 2 of the upper flow channel network respectively. n The terminal flow channels correspond to 2 n The step-down flow channel is L-shaped, the vertical portion of which is connected to the corresponding terminal flow channel of the upper flow channel network, and the horizontal portion is sandwiched between a group of terminal flow channels of the lower flow channel network and connected to the corresponding reaction chamber. The liquid outlet flow channel includes two corresponding channels connected to each reaction chamber. n The outlet liquid flow channel and 2 n The outlet of the total liquid outlet flow channel is arranged at the end surface of the chip body.

[0047] In the above chip, since each stepped flow channel cross section is sandwiched between a group of terminal flow channels of the lower flow channel network, the inner phase fluid flowing into the reaction chamber from the terminal flow channels of the upper flow channel network along the stepped flow channels is sandwiched on the left and right by the outer phase fluid flowing into the reaction chamber from a group of terminal flow channels of the lower flow channel network, so that the inner phase fluid can diffuse and mix with the outer phase fluid from both left and right directions, thereby accelerating the mixing efficiency of the inner phase fluid and the outer phase fluid.

[0048] Specifically, the first sheet material, the second sheet material and the third sheet material are all made of glass to improve biocompatibility and chemical stability.

[0049] In one embodiment, if Figure 1 As shown, the multi-channel microfluidic chip comprises:

[0050] The chip body includes a first sheet, a second sheet, and a third sheet stacked from top to bottom. In this embodiment, the first sheet, the second sheet, and the third sheet are all made of glass. Figure 1 Here, layer I refers to the first sheet material, layer II refers to the second sheet material, and layer III refers to the third sheet material.

[0051] Two layers of flow channel networks are arranged inside the chip body from top to bottom, the upper layer of the flow channel network is arranged between the first sheet and the second sheet, and the lower layer of the flow channel network is arranged between the second sheet and the third sheet;

[0052] The two-layer flow channel network is in a binary tree structure. In this embodiment, the upper layer flow channel network is provided with 8 terminal flow channels, and the lower layer flow channel network is provided with 16 terminal flow channels.

[0053] The terminal flow channel of the lower flow channel network is connected to a reaction chamber, and the terminal flow channel of the upper flow channel network is connected to the reaction chamber through a step-down flow channel, so as to make the fluid in the upper flow channel network and the fluid in the lower flow channel network mix and react in the reaction chamber;

[0054] There are 8 reaction chambers, and every two of the terminal flow channels of the lower flow channel network form a group and are connected to the same reaction chamber. There are 8 stepped-down flow channels, which respectively correspond to the 8 terminal flow channels of the upper flow channel network and the 8 reaction chambers. The stepped-down flow channels are L-shaped, and their vertical parts are connected to the corresponding terminal flow channels of the upper flow channel network, and their horizontal parts are sandwiched between a group of terminal flow channels of the lower flow channel network and are connected to the corresponding reaction chambers.

[0055] The root channel of the upper channel network is connected to a first liquid inlet channel 1 penetrating to the surface of the chip body, the root channel of the lower channel network is connected to a second liquid inlet channel 2 penetrating to the surface of the chip body, and the reaction chamber is connected to a liquid outlet channel penetrating to the surface of the chip body;

[0056] The liquid outlet flow channel includes 8 liquid outlet flow channels corresponding to and connected to each reaction chamber, and a total liquid outlet flow channel 3 that aggregates the 8 liquid outlet flow channels, and the outlet of the total liquid outlet flow channel is arranged on the end surface of the chip body;

[0057] Wherein, the width of the terminal flow channels of the upper layer flow channel network is 200 μm, and the width of the terminal flow channels of the lower layer flow channel network is 400 μm.

[0058] The present invention also provides a method for preparing curcumin-loaded nanoparticles using the multi-channel microfluidic chip, as shown in the following examples:

[0059] <Example 1>

[0060] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0061] Step 1: dissolve 10 mg of zein and 0.25 mg of curcumin in 1 mL of 80% ethanol, and disperse by ultrasonic for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0062] Step 2: Using the mixed alcohol solution of zein and curcumin in step 1 as the inner phase and the deionized water solution as the outer phase, the inner phase is introduced into the first liquid inlet channel of the aforementioned multi-channel microfluidic chip at an inner phase flow rate of 8 mL / h, and the outer phase is introduced into the second liquid inlet channel of the multi-channel microfluidic chip at an outer phase flow rate of 160 mL / h, and a dispersion of zein nanoparticles loaded with curcumin is obtained from the liquid outlet channel of the multi-channel microfluidic chip;

[0063] Step 3: Place the curcumin-loaded zein nanoparticle dispersion prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0064] The particle size distribution diagram of the loaded curcumin nanoparticles prepared in Example 1 is shown in the attached figure. Figure 2 As shown, it is a single peak with a particle size of about 177 nm, a polydispersity index (PDI) of 0.072, and a curcumin loading rate of 99.87±0.51%.

[0065] The curcumin loading rate is obtained by the following method: dissolve the curcumin standard in anhydrous ethanol and dilute it to 0-10 μg / mL, and test the absorbance at 426 nm to make a standard curve. Centrifuge the loaded curcumin nanoparticles in a 10kDa ultrafiltration centrifuge tube at 4000rpm for 30min, dilute the filtrate 10 times with anhydrous ethanol, test the absorbance at 426nm, and substitute it into the standard curve to obtain the mass of free curcumin. The amount of loaded curcumin is obtained by subtracting the mass of free curcumin from the total amount of curcumin taken when preparing the loaded curcumin nanoparticles, and the loading rate is the proportion of the amount of loaded curcumin in the total amount of curcumin.

[0066] The curcumin loading rate was obtained by the above method in the following examples and comparative examples.

[0067] <Example 2>

[0068] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0069] Step 1, dissolving 10 mg of zein and 0.5 mg of curcumin in 1 mL of 80% ethanol, and ultrasonically dispersing for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0070] Step 2: Using the mixed alcohol solution of zein and curcumin in step 1 as the inner phase and the deionized water solution as the outer phase, the inner phase is introduced into the first liquid inlet channel of the aforementioned multi-channel microfluidic chip at an inner phase flow rate of 8 mL / h, and the outer phase is introduced into the second liquid inlet channel of the multi-channel microfluidic chip at an outer phase flow rate of 160 mL / h, and a dispersion of zein nanoparticles loaded with curcumin is obtained from the liquid outlet channel of the multi-channel microfluidic chip;

[0071] Step 3: Place the dispersion of the curcumin nanoparticles prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0072] The particle size distribution diagram of the loaded curcumin nanoparticles prepared in Example 2 is shown in the attached figure. Figure 3 As shown, it is a single peak with a particle size of about 182.2 nm, a polydispersity index (PDI) of 0.115, and a curcumin loading rate of 99.16±0.78%.

[0073] <Example 3>

[0074] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0075] Step 1: 10 mg of zein and 1 mg of curcumin were ultrasonically dispersed in 1 mL of 80% ethanol for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0076] Step 2: Using the mixed alcohol solution of zein and curcumin in step 1 as the inner phase and the deionized water solution as the outer phase, the inner phase is introduced into the first liquid inlet channel of the aforementioned multi-channel microfluidic chip at an inner phase flow rate of 8 mL / h, and the outer phase is introduced into the second liquid inlet channel of the multi-channel microfluidic chip at an outer phase flow rate of 160 mL / h, and a dispersion of zein nanoparticles loaded with curcumin is obtained from the liquid outlet channel of the multi-channel microfluidic chip;

[0077] Step 3: Place the dispersion of the curcumin nanoparticles prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0078] The particle size distribution diagram of the curcumin nanoparticles prepared in Example 3 is shown in the attached figure. Figure 4 As shown, it is a single peak with a particle size of about 185 nm, a polydispersity index (PDI) of 0.105, and a curcumin loading rate of 99.09±0.07%.

[0079] <Comparative Example 1>

[0080] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0081] Step 1: dissolve 10 mg of zein and 0.25 mg of curcumin in 1 mL of 80% ethanol, and disperse by ultrasonic for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0082] Step 2, dispersing the mixed alcohol solution of zein and curcumin in step 1 into 20 mL of deionized water to obtain a dispersion of curcumin nanoparticles;

[0083] Step 3: Place the dispersion of the curcumin nanoparticles prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0084] The particle size distribution diagram of the curcumin nanoparticles prepared in Comparative Example 1 is shown in the attached figure. Figure 5 As shown, it was multimodal, the polydispersity index (PDI) was 0.271, and the curcumin loading rate was 96.68±0.52%.

[0085] <Comparative Example 2>

[0086] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0087] Step 1, dissolving 10 mg of zein and 0.5 mg of curcumin in 1 mL of 80% ethanol, and ultrasonically dispersing for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0088] Step 2, dispersing the mixed alcohol solution of zein and curcumin in step 1 into 20 mL of deionized water to obtain a dispersion of curcumin nanoparticles;

[0089] Step 3: Place the dispersion of the curcumin nanoparticles prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0090] The particle size distribution diagram of the loaded curcumin nanoparticles prepared in Comparative Example 2 is shown in the attached figure. Figure 6 As shown, it is multi-peaked, the polydispersity index (PDI) is 0.505, and the curcumin loading rate is 92.77±0.73%.

[0091] <Comparative Example 3>

[0092] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0093] Step 1, dissolving 10 mg of zein and 1 mg of curcumin in 1 mL of 80% ethanol, and ultrasonically dispersing for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0094] Step 2, dispersing the mixed alcohol solution of zein and curcumin in step 1 into 20 mL of deionized water to obtain a dispersion of curcumin nanoparticles;

[0095] Step 3: Place the dispersion of the curcumin nanoparticles prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0096] The particle size distribution diagram of the loaded curcumin nanoparticles prepared in Comparative Example 3 is shown in the attached figure. Figure 7 As shown, it is multimodal, the polydispersity index (PDI) is 0.512, and the curcumin loading rate is 87.67±0.84%.

[0097] From the above embodiments and comparative examples, the difference between Example 1 and Comparative Example 1 is that a multichannel microfluidic chip is used in Example 1, while in Comparative Example 1, the mixed alcohol solution is directly dispersed in deionized water. According to the particle size distribution diagrams of the product of Example 1 and the product of Comparative Example 1, the particle size distribution of the product of Example 1 is unimodal and has a high peak intensity, indicating that the particle size distribution is relatively concentrated and the size and morphology of the prepared nanoparticles are relatively uniform, while the particle size distribution of the product of Comparative Example 1 is wide and multimodal, and the peak intensity is low, indicating that the particle size distribution is relatively extensive and not concentrated, and the size and morphology of the prepared nanoparticles are relatively diverse. The particle size of the product of Example 1 is about 177 nm, the polydispersity index (PDI) is 0.072, and the curcumin loading rate is 99.87±0.51%, while the polydispersity index (PDI) of the product of Comparative Example 1 is 0.271, and the curcumin loading rate is 96.68±0.52%, which further proves that the product of Example 1 has better morphological uniformity and higher loading rate than Comparative Example 1.

[0098] The difference between Example 2 and Comparative Example 2 is that a multi-channel microfluidic chip is used in Example 2, while in Comparative Example 2, the mixed alcohol solution is directly dispersed into deionized water. According to the particle size distribution diagrams of the products of Example 2 and Comparative Example 2, the particle size distribution of the product of Example 2 is unimodal and has a high peak intensity, indicating that the particle size distribution is more concentrated and the size and morphology of the prepared nanoparticles are more uniform, while the particle size distribution of the product of Comparative Example 2 is wide and multimodal, with low peak intensity and some particle sizes reach more than 1000 nm, indicating that the particle size distribution is relatively wide and not concentrated, the size and morphology of the prepared nanoparticles are more diverse, and some particles are agglomerated, resulting in a significant increase in particle size, resulting in a decrease in the yield of nanoparticles. The particle size of the product of Example 2 is about 182.2 nm, the polydispersity index (PDI) is 0.115, and the curcumin loading rate is 99.16±0.78%, while the polydispersity index (PDI) of the product of Comparative Example 2 is 0.505, and the curcumin loading rate is 92.77±0.73%, which further proves that the product of Example 1 has better morphological uniformity and higher loading rate than Comparative Example 1.

[0099] The difference between Example 3 and Comparative Example 3 is that a multi-channel microfluidic chip is used in Example 3, while in Comparative Example 3, the mixed alcohol solution is directly dispersed in deionized water. According to the particle size distribution diagrams of the product of Example 3 and the product of Comparative Example 3, the particle size distribution of the product of Example 3 is unimodal and has a high peak intensity, indicating that the particle size distribution is more concentrated and the size and morphology of the prepared nanoparticles are more uniform, while the particle size distribution of the product of Comparative Example 3 is wide and multimodal, with low peak intensity and some particle sizes reach more than 1000 nm, indicating that the particle size distribution is relatively wide and not concentrated, the size and morphology of the prepared nanoparticles are more diverse, and some particles are agglomerated, resulting in a significant increase in the particle size, resulting in a decrease in the yield of nanoparticles. The particle size of the product of Example 3 is about 185 nm, the polydispersity index (PDI) is 0.105, and the curcumin loading rate is 99.09±0.07%, while the polydispersity index (PDI) of the product of Comparative Example 3 is 0.512, and the curcumin loading rate is 87.67±0.84%, which further proves that the product of Example 1 has better morphological uniformity and higher loading rate than Comparative Example 1.

[0100] <Comparative Example 4>

[0101] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0102] Step 1: 10 mg of zein and 1 mg of curcumin were ultrasonically dispersed in 1 mL of 80% ethanol for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0103] Step 2: Using the mixed alcohol solution of zein and curcumin in step 1 as the inner phase and the deionized water solution as the outer phase, the inner phase is introduced into the first liquid inlet channel of the multichannel microfluidic chip in Example 1 at an inner phase flow rate of 3 mL / h, and the outer phase is introduced into the second liquid inlet channel of the multichannel microfluidic chip at an outer phase flow rate of 60 mL / h, and a dispersion of zein nanoparticles loaded with curcumin is obtained from the liquid outlet channel of the multichannel microfluidic chip;

[0104] Step 3: Place the dispersion of the curcumin nanoparticles prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0105] The particle size distribution diagram of the loaded curcumin nanoparticles prepared in Comparative Example 4 is shown in the attached figure. Figure 8 As shown, it is a single peak with a particle size of about 2613 nm and a polydispersity index (PDI) of 0.422.

[0106] <Comparative Example 5>

[0107] The preparation method of curcumin-loaded nanoparticles comprises the following steps:

[0108] Step 1: 10 mg of zein and 1 mg of curcumin were ultrasonically dispersed in 1 mL of 80% ethanol for 5 min to obtain a mixed alcohol solution of zein and curcumin;

[0109] Step 2: Using the mixed alcohol solution of zein and curcumin in step 1 as the inner phase and the deionized water solution as the outer phase, the inner phase is introduced into the first liquid inlet channel of the multichannel microfluidic chip in Example 1 at an inner phase flow rate of 30 mL / h, and the outer phase is introduced into the second liquid inlet channel of the multichannel microfluidic chip at an outer phase flow rate of 300 mL / h, and a dispersion of zein nanoparticles loaded with curcumin is obtained from the liquid outlet channel of the multichannel microfluidic chip;

[0110] Step 3: Place the dispersion of the curcumin nanoparticles prepared in step 2 in a 10KD dialysis bag for 48 hours, collect the suspension in the dialysis bag, freeze-dry, and collect the particle sample.

[0111] The particle size distribution diagram of the loaded curcumin nanoparticles prepared in Comparative Example 5 is shown in the attached figure. Fig. 9 As shown, it is multi-peaked, the particle size is about 42.6nm, and the polydispersity index (PDI) is 0.917.

[0112] From the above embodiments and comparative examples, the difference between Example 3 and Comparative Example 4 is that in Example 3, the internal phase flow rate is 8 mL / h and the external phase flow rate is 160 mL / h, while in Comparative Example 4, the internal phase flow rate is 3 mL / h and the external phase flow rate is 60 mL / h. According to the particle size distribution diagrams of the product of Example 3 and the product of Comparative Example 4, the particle size distribution of the product of Example 3 is unimodal and has high peak intensity, indicating that the particle size distribution is relatively concentrated and the prepared nanoparticles are of uniform size and shape. Although the particle size distribution of the product of Comparative Example 4 is unimodal, the particle size value reaches 2613 nm and the polydispersity index (PDI) is 0.422, which indicates that the large-scale agglomeration of the particles leads to a substantial increase in the particle size, and the loaded curcumin nanoparticles can no longer be prepared. This also shows that the internal phase flow rate and the external phase flow rate are too slow to prepare the loaded curcumin nanoparticles.

[0113] The difference between embodiment 3 and comparative example 5 is that in embodiment 3, internal phase flow rate is 8mL / h, and external phase flow rate is 160mL / h, and in comparative example 5, internal phase flow rate is 30mL / h, and external phase flow rate is 300mL / h, according to the particle size distribution diagram of embodiment 3 product and comparative example 5 product, the particle size distribution of embodiment 3 product is unimodal, and peak intensity is high, indicating that particle size distribution is more concentrated, and the size form of the prepared nanoparticles is more uniform, while the particle size distribution of comparative example 5 product is wide and multimodal, and peak intensity is low and part of particle size reaches more than 1000nm, indicating that particle size distribution is more widely not concentrated, and the size form of the prepared nanoparticles is more diverse, and some particles have agglomerated and caused particle size to be greatly improved, causing nanoparticle yield to be reduced. The main peak particle diameter of comparative example 5 product is about 42.6nm, and polydispersity index (PDI) is 0.917, which also illustrates that the internal phase flow rate is too fast and can not prepare uniform morphology, and the loaded curcumin nanoparticles with high loading rate.

[0114] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and implementation modes. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and the illustrations shown and described herein.

Claims

1. A multi-channel microfluidic chip, characterized in that: include: The chip body has at least two layers of flow channel networks arranged from top to bottom, both of which are in a binary tree structure, the terminal flow channels of the lower flow channel network are connected to the reaction chamber, and the terminal flow channels of the upper flow channel network are connected to the reaction chamber through a descending flow channel, so as to make the fluid in the upper flow channel network and the fluid in the lower flow channel network mix and react in the reaction chamber; The root channel of the upper channel network is connected to a first liquid inlet channel that penetrates the surface of the chip body, the root channel of the lower channel network is connected to a second liquid inlet channel that penetrates the surface of the chip body, and the reaction chamber is connected to a liquid outlet channel that penetrates the surface of the chip body; The width of the distal flow channels of the upper layer flow channel network is 150 to 250 μm, and the width of the distal flow channels of the lower layer flow channel network is 290 to 500 μm.

2. The multi-channel microfluidic chip according to claim 1, characterized in that: The chip body includes a first sheet, a second sheet and a third sheet stacked from top to bottom, the upper channel network is arranged between the first sheet and the second sheet, and the lower channel network, the reaction chamber and the liquid outlet channel are arranged between the second sheet and the third sheet.

3. The multi-channel microfluidic chip according to claim 1, characterized in that: The terminal flow channel of the upper flow channel network is provided with 2 n The terminal flow channel of the lower flow channel network is provided with 2 n+1 , n>0, the reaction chamber is provided with 2 n Each of the two distal flow channels of the lower layer flow channel network is connected to the same reaction chamber as a group, and the step-down flow channel is provided with 2 n and correspond to the 2 of the upper flow channel network respectively. n The terminal flow channels correspond to 2 n The step-down flow channel is L-shaped, the vertical portion of which is connected to the corresponding terminal flow channel of the upper flow channel network, and the horizontal portion is sandwiched between a group of terminal flow channels of the lower flow channel network and connected to the corresponding reaction chamber. The liquid outlet flow channel includes two corresponding channels connected to each reaction chamber. n The outlet liquid flow channel and 2 n The outlet of the total liquid outlet flow channel is arranged at the end surface of the chip body.

4. The multi-channel microfluidic chip according to claim 2, characterized in that: The first sheet material, the second sheet material and the third sheet material are all made of glass.

5. A method for preparing curcumin-loaded nanoparticles using the multi-channel microfluidic chip according to any one of claims 1 to 4, characterized in that: The following steps are involved: Step 1, dissolving zein and curcumin in ethanol, and dispersing by ultrasonication to obtain a mixed alcohol solution of zein and curcumin; Step 2: using the mixed alcohol solution as the inner phase and the ultrapure aqueous solution as the outer phase, passing the inner phase into the first liquid inlet channel of the multi-channel microfluidic chip, passing the outer phase into the second liquid inlet channel of the multi-channel microfluidic chip, and obtaining a dispersion of zein nanoparticles loaded with curcumin from the liquid outlet channel of the multi-channel microfluidic chip; Step 3: placing the zein nanoparticle dispersion loaded with curcumin in a dialysis bag for dialysis for a certain period of time, and freeze-drying to obtain the curcumin nanoparticles loaded with curcumin.

6. The method for preparing loaded curcumin nanoparticles as claimed in claim 5, characterized in that: In step 1, the concentration of zein is 5-40 mg / mL, the mass ratio of curcumin to zein is 1:5-50, and the volume fraction of ethanol is 60-95%.

7. The method for preparing the loaded curcumin nanoparticles according to claim 5, wherein: In step 2, the flow rate of the inner phase is 4-24 mL / h, and the flow rate of the outer phase is 80-300 mL / h.

8. The method for preparing the loaded curcumin nanoparticles according to claim 5, wherein: A 10KD dialysis bag was used and the dialysis time was 24 to 72 hours.

9. The curcumin-loaded nanoparticles prepared by the method according to any one of claims 5 to 8.

10. Use of the curcumin-loaded nanoparticles as claimed in claim 9 in loading and delivering in food, medicine and health products.

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