Dual-load food-grade janus particles, methods of making and using the same

Abstract

The application discloses a preparation method of double-load food-grade Janus particles, and comprises the following steps: S1, dissolving zein, shellac, curcumin and resveratrol in ethanol, and performing ultrasonic dispersion to obtain a mixed alcohol solution of zein, shellac, curcumin and resveratrol; adding Tween 80 into deionized water, and performing ultrasonic dispersion to obtain a Tween 80 aqueous solution; S2, taking the mixed alcohol solution as an inner phase, taking the Tween 80 aqueous solution as an outer phase, and passing through a coaxial flow focusing type chip to obtain a dispersion liquid of the double-load food-grade Janus particles in the Tween 80 aqueous solution; and S3, placing the dispersion liquid into a dialysis bag to dialyze for a certain time, and performing freeze-drying to obtain the Janus particles. The preparation method has low cost, good particle loading effect and easy scale production. The application further provides the double-load food-grade Janus particles and application of the double-load food-grade Janus particles in loading and delivering curcumin and resveratrol. The Janus particles have the beneficial effect of high loading rate.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of new particle materials. More particularly, the present application relates to a dual-loaded food-grade Janus particle and a preparation method and application thereof. BACKGROUND

[0002] Curcumin and Resveratrol are both bioactive compounds of natural origin, with antioxidant, anti-inflammatory, anticancer and cardiovascular protection, and other health benefits. However, due to the low solubility in water, the bioavailability is not high, which limits the effective absorption of the human body.

[0003] In order to overcome these limitations and achieve synergistic therapeutic effect, researchers have studied the use of nanoparticles, liposomes, Pickering emulsion and other carriers to simultaneously load the two active substances. A co-loaded zein particle of curcumin-resveratrol uses nanotechnology to load two functional factors in biodegradable proteins. This carrier can protect the drug from the environment, increase the solubility, and improve the loading efficiency of functional factors. In addition, using liposome preparation technology, a double or multi-layer vesicle structure can be formed to encapsulate curcumin and resveratrol, significantly improving their water solubility and bioavailability, and delivering drugs to the cell interior by mechanisms such as cell membrane fusion.

[0004] Janus particles, as a material with double or multi-region different physical and chemical properties, have significant advantages in loading two or more active substances. Its asymmetric characteristics allow different active ingredients (such as drugs, enzymes, catalysts, etc.) to be loaded on both sides or both sides, thereby achieving spatial isolation and avoiding incompatible reactions or non-target effects between them, allowing each substance to be released independently or synergistically under specific conditions. In addition, due to the different hydrophilicity, charge distribution or other characteristics of each region, the release rate and order of each loaded substance can be precisely controlled according to the needs. However, it is difficult to achieve high loading rate of dual loading when loading two target active substances. SUMMARY

[0005] An object of the present application is to solve at least the above problems and to provide at least the advantages to be described later.

[0006] In order to achieve these objects and other advantages in accordance with the present application, a preparation method of a dual-loaded food-grade Janus particle is provided, comprising the following steps:

[0007] S1, dissolving zein, shellac, curcumin and resveratrol in ethanol, ultrasonic dispersion, to obtain a mixed alcohol solution of zein, shellac, curcumin and resveratrol;

[0008] Tween 80 is added to deionized water, ultrasonic dispersion, to obtain Tween 80 aqueous solution;

[0009] S2, with mixed alcohol solution as the inner phase, Tween 80 aqueous solution as the outer phase, coaxial flow focusing chip is passed, to obtain the dispersion of double-loaded food-grade Janus particles in Tween 80 aqueous solution;

[0010] S3, the dispersion is placed in the dialysis bag for a certain time, and the suspension in the dialysis bag is collected and freeze-dried to obtain the Janus particles.

[0011] Preferably, the concentration of zein and shellac in step S1 is 6-96 mg / mL, the mass ratio of curcumin and resveratrol to zein and shellac is 1:10-80; the mass ratio of curcumin and resveratrol is 4:1-16, and the ethanol is 80-95% ethanol.

[0012] Preferably, the mass-volume ratio of Tween 80 to deionized water is 1g:100mL.

[0013] Preferably, the coaxial flow focusing chip is passed at 70-95 DEG C, with an inner phase flow rate of 1-3 mL / h and an outer phase flow rate of 15-50 mL / h.

[0014] Preferably, a dialysis bag with a molecular weight of 10KD is selected, and the dialysis time is 24-72h.

[0015] A double-loaded food-grade Janus particle is provided.

[0016] Preferably, the double-loaded food-grade Janus particle is dumbbell-shaped or snowman-shaped, with zein at one end and shellac at the other end.

[0017] In the double-loaded food-grade Janus particle, curcumin tends to be inside the shellac end of the food-grade Janus particle, and resveratrol tends to be inside the zein end.

[0018] The application provides a double-loaded food-grade Janus particle for loading and delivering curcumin and resveratrol in food, medicine and health products.

[0019] The present application at least includes the following beneficial effects:

[0020] First, the present application uses natural biological macromolecule zein and shellac as the construction raw material of Janus particles, which is green and natural. The two hemispheres of the prepared Janus particles are zein and shellac, respectively, so that the Janus particles have the properties of both materials.

[0021] Secondly, the present application prepares particles by a coaxial flow focusing type chip of microfluidics, and the preparation method is simple, easy to operate, and can be continuously and mass-produced.

[0022] Thirdly, in the present application, a series of Janus particles can be obtained by regulating the concentration of zein and shellac, the flow rate of inner and outer phases, and the volume concentration of ethanol, so as to precisely regulate the particle morphology and size and stabilize emulsions of different sizes and types.

[0023] Fourthly, the raw materials for preparing the Janus particles are natural biological macromolecules, which are non-toxic, harmless, environmentally friendly, and biodegradable green materials, so that the Janus particles can be applied in the fields of food or medicine.

[0024] Other advantages, objects, and features of the present application will be partly embodied in the following description, and will be partly understood by those skilled in the art through research and practice of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 A scanning electron microscope image of the double-loaded food-grade Janus particles prepared in Example 1 of the present application;

[0026] Figure 2 A scanning electron microscope image of the double-loaded food-grade Janus particles prepared in Example 2 of the present application;

[0027] Figure 3 A scanning electron microscope image of the double-loaded food-grade Janus particles prepared in Example 3 of the present application;

[0028] Figure 4 A scanning electron microscope image of the particles prepared in Comparative Example 1 of the present application;

[0029] Figure 5 A scanning electron microscope image of the particles prepared in Comparative Example 2 of the present application;

[0030] Figure 6 A scanning electron microscope image of the particles prepared in Comparative Example 3 of the present application. DETAILED DESCRIPTION

[0031] The present application will be further described in detail below with reference to the accompanying drawings, so that those skilled in the art can implement the present application according to the description.

[0032] It should be noted that the experimental methods in the following embodiments are conventional methods, and the reagents and materials can be obtained from commercial channels unless otherwise specified.

[0033] <Example 1>

[0034] The preparation method of the double-loaded food-grade Janus particles comprises the following steps:

[0035] Step one, 5 mg of zein, 15 mg of shellac, 1 mg of curcumin and 1 mg of resveratrol were dissolved in 1 mL of 90% ethanol, and ultrasonic dispersion was performed for 5 min to obtain a mixed alcohol solution of zein, shellac, curcumin and resveratrol;

[0036] 1 g of Tween 80 was added to 100 mL of deionized water, and ultrasonic dispersion was performed for 5 min to obtain a 1% Tween 80 aqueous solution;

[0037] Step two, the mixed alcohol solution of zein, shellac, curcumin and resveratrol in step one was used as the inner phase, and the Tween 80 aqueous solution was used as the outer phase, and a coaxial flow focusing type chip was passed in at an inner phase flow rate of 1.5 mL / h and an outer phase flow rate of 30 mL / h at 80°C to obtain a dispersion of double-loaded food-grade Janus particles in the Tween 80 aqueous solution;

[0038] Step three, the dispersion of double-loaded food-grade Janus particles in the Tween 80 aqueous solution prepared in step two was placed in a 10KD dialysis bag for dialysis for 48 h, the suspension in the dialysis bag was collected, freeze-dried, and the particle sample was collected.

[0039] The scanning electron microscope image of the Janus particles prepared in Example 1 is shown in FIG. 1. Figure 1 The Janus particles are in the shape of a snowman, with a particle size of about 700 nm, a curcumin loading rate of 92.05±0.56%, a resveratrol loading rate of 98.71±0.78%, and a total loading amount of 8.67±0.15%.

[0040] <Example 2>

[0041] The preparation method of the double-loaded food-grade Janus particles comprises the following steps:

[0042] Step one, 10 mg of zein, 10 mg of shellac, 1 mg of curcumin and 1 mg of resveratrol were dissolved in 1 mL of 90% ethanol, and ultrasonic dispersion was performed for 5 min to obtain a mixed alcohol solution of zein, shellac, curcumin and resveratrol;

[0043] 1 g of Tween 80 was added to 100 mL of deionized water, and ultrasonic dispersion was performed for 5 min to obtain a 1% Tween 80 aqueous solution;

[0044] Step 2: Using the mixed alcoholic solution of zein, shellac, curcumin and resveratrol from Step 1 as the inner phase and Tween 80 aqueous solution as the outer phase, the solution is passed into a coaxial flow focusing chip at 80°C at an inner phase flow rate of 1.5 mL / h and an outer phase flow rate of 30 mL / h to obtain a dispersion of dual-loaded food-grade Janus particles in Tween 80 aqueous solution.

[0045] Step 3: The dispersion of the dual-loaded food-grade Janus particles obtained in Step 2 in Tween 80 aqueous solution was placed in a 10KD dialysis bag and dialyzed for 48 hours. The suspension in the dialysis bag was collected, freeze-dried, and the particle sample was collected.

[0046] Scanning electron microscope images of the Janus particles prepared in Example 2 are attached. Figure 2 As shown, the particles are dumbbell-shaped with a diameter of approximately 600 nm, a curcumin loading rate of 92.53±0.26%, a resveratrol loading rate of 97.94±0.58%, and a total loading of 8.65±0.13%.

[0047] <Example 3>

[0048] A method for preparing dual-loaded food-grade Janus granules includes the following steps:

[0049] Step 1: Dissolve 15mg zein, 5mg shellac, 1mg curcumin and 1mg resveratrol in 1mL of 90% ethanol and sonicate for 5min to obtain a mixed alcoholic solution of zein, shellac, curcumin and resveratrol.

[0050] Add 1g of Tween 80 to 100mL of deionized water and sonicate for 5min to obtain a 1% Tween 80 aqueous solution;

[0051] Step 2: Using the mixed alcoholic solution of zein, shellac, curcumin and resveratrol from Step 1 as the inner phase and Tween 80 aqueous solution as the outer phase, the solution is passed into a coaxial flow focusing chip at 80°C at an inner phase flow rate of 1.5 mL / h and an outer phase flow rate of 30 mL / h to obtain a dispersion of dual-loaded food-grade Janus particles in Tween 80 aqueous solution.

[0052] Step 3: The dispersion of the dual-loaded food-grade Janus particles obtained in Step 2 in Tween 80 aqueous solution was placed in a 10KD dialysis bag and dialyzed for 48 hours. The suspension in the dialysis bag was collected, freeze-dried, and the particle sample was collected.

[0053] Scanning electron microscope (SEM) images of the Janus particles prepared in Example 3 are attached. Figure 3 As shown, the particles are snowman-shaped with a diameter of approximately 200 nm. The curcumin loading rate is 93.36±0.16%, the resveratrol loading rate is 99.29±0.47%, and the total loading rate is 8.75±0.17%.

[0054] <Comparative Example 1>

[0055] Step 1: Dissolve 20 mg of zein, 1 mg of curcumin and 1 mg of resveratrol in 1 mL of 90% ethanol and sonicate for 5 min to obtain a mixed alcoholic solution of zein, curcumin and resveratrol.

[0056] Step 2: Using the mixed alcoholic solution of zein, curcumin, and resveratrol from Step 1 as the inner phase and deionized water as the outer phase, the solution is passed into a coaxial flow focusing chip at 80°C at an inner phase flow rate of 1.5 mL / h and an outer phase flow rate of 30 mL / h to obtain a dispersion.

[0057] Step 3: Place the dispersion obtained in Step 2 into a 10KD dialysis bag and dialyze for 48 hours. Collect the suspension in the dialysis bag, freeze-dry it, and collect the particulate sample.

[0058] Scanning electron microscope images of the particles prepared in Comparative Example 1 are attached. Figure 4 As shown, the particles are spherical with a diameter of approximately 180 nm, and have a curcumin loading rate of 92.25±0.14%, a resveratrol loading rate of 68.97±0.85%, and a total loading of 7.32±0.28%.

[0059] <Comparative Example 2>

[0060] Step 1: Dissolve 20mg shellac, 1mg curcumin and 1mg resveratrol in 1mL of 90% ethanol and sonicate for 5min to obtain a mixed alcoholic solution of shellac, curcumin and resveratrol.

[0061] Step 2: Using the mixed alcoholic solution of shellac, curcumin, and resveratrol from Step 1 as the inner phase and deionized water as the outer phase, the solution is passed into a coaxial flow focusing chip at 80°C at an inner phase flow rate of 1.5 mL / h and an outer phase flow rate of 30 mL / h to obtain a dispersion.

[0062] Step 3: Place the dispersion obtained in Step 2 into a 10KD dialysis bag and dialyze for 48 hours. Collect the suspension in the dialysis bag, freeze-dry it, and collect the particulate sample.

[0063] Scanning electron microscope (SEM) images of the particles prepared in Comparative Example 2 are attached. Figure 5 As shown, the particles are spherical with a diameter of approximately 145 nm, and have a curcumin loading rate of 92.93±0.08%, a resveratrol loading rate of 60.44±0.45%, and a total loading of 6.97±0.19%.

[0064] <Comparative Example 3>

[0065] Step one, 10 mg of zein, 10 mg of shellac, 1 mg of curcumin and 1 mg of resveratrol were dissolved in 1 mL of 90% ethanol, ultrasonic dispersion for 5 min, to obtain a mixed alcohol solution of zein, shellac, curcumin and resveratrol;

[0066] Step two, the mixed alcohol solution of zein, shellac, curcumin and resveratrol in step one was used as the inner phase, and deionized water was used as the outer phase, and the coaxial flow focusing chip was passed at 80°C with an inner phase flow rate of 1.5 mL / h and an outer phase flow rate of 30 mL / h to obtain a dispersion;

[0067] Step three, the dispersion prepared in step two was placed in a 10KD dialysis bag and dialyzed for 48h, the suspension in the dialysis bag was collected and freeze-dried to collect the granular sample.

[0068] The scanning electron microscope image of the granules prepared in Comparative Example 3 is shown in FIG. 1, which is spherical with a particle size of about 4500 nm, the curcumin loading rate is 93.04±0.21%, the resveratrol loading rate is 72.28±0.31%, and the total loading capacity is 7.51±0.24%. Figure 6

[0069] Table 1 loading rate and total loading capacity

[0070]

[0071] Comparative Examples 1-3, the loading rate and total loading capacity of curcumin and resveratrol are significantly improved, indicating that the composite Janus particles can significantly improve the loading effect of the two functional factors. This is because zein can self-assemble into nanoparticles through non-covalent interactions such as hydrophobic interaction and hydrogen bonding, and curcumin and resveratrol, as hydrophobic polyphenolic compounds, can be physically loaded by embedding into the hydrophobic core of zein nanoparticles. Shellac is a natural resin containing various phenolic and acidic compounds, which can form stable complexes with zein in a similar way, thereby achieving encapsulation of hydrophobic functional factors. The loading effect of the composite nanoparticles is relatively improved, and the loading rate of the composite Janus particles on curcumin and resveratrol is significantly improved, especially resveratrol, which may be due to the presence of surfactant in the outer phase, which can delay the nucleation process of the inner phase polymer and functional components, helping to encapsulate the functional factors, so that the total loading capacity of the double-loaded composite Janus particles is significantly improved. In addition, different morphologies of composite Janus particles can stabilize different types of emulsions (oil-in-water or water-in-oil), and the cell uptake effect will also be different. And Comparative Examples 1-3 cannot form composite Janus particles, even if Tween is added in Comparative Examples 1 and 2, composite Janus particles cannot be formed, therefore, they also do not have the structural basis for simultaneously loading curcumin and resveratrol. ​

[0072] While embodiments of the application have been disclosed in connection with the above specification and drawings this description is not intended to limit the scope of the application and many modifications, enhancements, alternatives, and variations will become apparent to those skilled in the art from this disclosure. Accordingly, it is expressly intended that the description should not limit the application as claimed but rather the only limitation placed on the scope of the application be the scope of the claims as set out below and equivalents thereof.

Claims

1. A method of preparing dual-loaded food-grade Janus particles, characterized in that, The method comprises the following steps: S1, dissolving zein, shellac, curcumin and resveratrol in ethanol, ultrasonic dispersion, obtaining a mixed alcohol solution of zein, shellac, curcumin and resveratrol; Tween 80 is added to deionized water, ultrasonic dispersion, obtaining a Tween 80 aqueous solution; S2, taking the mixed alcohol solution as the inner phase, the Tween 80 aqueous solution as the outer phase, passing through a coaxial flow focusing chip, obtaining a dispersion of the double-loaded food-grade Janus particles in the Tween 80 aqueous solution; S3, placing the dispersion in a dialysis bag for dialysis for a certain time, collecting the suspension in the dialysis bag and freeze-drying, obtaining the Janus particles.

2. The method of making dual-load food-grade Janus particles of claim 1, wherein, In step S1, the concentrations of zein and shellac are both 6-96 mg / mL, the mass ratio of curcumin and resveratrol to zein and shellac is 1:10-80, the mass ratio of curcumin and resveratrol is 4:1-16, and the ethanol is 80-95% ethanol.

3. The method of making dual-load food-grade Janus particles of claim 2, wherein, The mass-volume ratio of Tween 80 to deionized water is 1g:100 mL.

4. The method of making dual-load food-grade Janus particles of claim 3, wherein, The coaxial flow focusing chip is passed at 70-95°C, with an inner phase flow rate of 1-3 mL / h and an outer phase flow rate of 15-50 mL / h.

5. The method of making dual-load food-grade Janus particles of claim 4, wherein, A dialysis bag with a molecular weight of 10 KD is selected, and the dialysis time is 24-72 h.

6. The double-loaded food-grade Janus particles prepared by the preparation method according to any one of claims 1-5.

7. The dual-load food-grade Janus particle of claim 6, wherein, The double-loaded food-grade Janus particles are dumbbell-shaped or snowman-shaped, with zein at one end and shellac at the other end. In the double-loaded food-grade Janus particles, curcumin tends to be inside the shellac end of the food-grade Janus particles, and resveratrol tends to be inside the zein end.

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