Preparation method and application of controlled-release antibacterial nanofiber hydrogel film

Abstract

The invention belongs to the technical field related to food processing, and particularly relates to a preparation method and application of a controlled-release antibacterial nanofiber hydrogel film. Gelatin and zein are used as basic raw materials, microporous microspheres are formed by mixing and utilizing a supercritical effect, then the microspheres are used as carriers, EDC / NHS is used for activating groups on the surfaces of the carriers, different perillaldehyde is effectively loaded, the overall stability and antibacterial performance are kept, meanwhile, the insurance effect is outstanding, and the nano-fiber hydrogel film has a good application prospect. A novel and effective antibacterial package is provided for the food industry.

Description

Technical Field

[0001] The invention belongs to the technical field related to food processing, and specifically relates to a preparation method of a controlled-release antibacterial nanofiber hydrogel membrane and application thereof. Background Art

[0002] Mutton is rich in nutrients and is loved by people all over the world for its unique flavor and high protein content. At present, the market is mainly composed of cold fresh meat, frozen meat, and hot fresh meat. Cold fresh mutton has gradually become the mainstream sales because of its more tender taste. However, during production and transportation, cold fresh mutton is easily contaminated by microorganisms and spoiled. Therefore, the main problem currently faced is to extend the shelf life of cold fresh mutton and improve product quality. It is imperative to seek a new and effective preservation technology.

[0003] Zein, chitosan, sodium alginate, starch, methylcellulose, etc. are commonly used carrier matrices for preparing edible films. Active films prepared from single biopolymers cannot meet the actual needs of food packaging and preservation due to their hydrophilic and easily degradable properties. This defect is often compensated by compounding different biopolymers to form efficient water vapor barriers and hydrophobic properties. It has been reported that chitosan or zein and gelatin can enhance interactions through electrostatic and hydrogen bonds between functional groups, which can avoid the phenomenon that single gelatin polymer films are rapidly decomposed when they come into contact with water. However, the above-mentioned films cannot continuously inhibit the reproduction of microorganisms during the storage of meat, resulting in the problem of "sudden release" of antibacterial substances in the films; since essential oils are volatile substances, existing studies have failed to control the release of essential oil active substances well, resulting in the problems of weak controlled release of antibacterial active substances, low antibacterial efficiency, and poor barrier properties in bio-based packaging films. Summary of the invention

[0004] In response to the above technical problems, the present invention provides a method for preparing a nanofiber hydrogel membrane that can effectively load perillaldehyde and effectively release controlled-release antibacterial agents. At the same time, the nanofiber hydrogel membrane exhibits significant antibacterial properties and stable preservation effects, which significantly extends the shelf life of fresh mutton and is suitable for the packaging and preservation of fresh meat such as mutton.

[0005] To this end, the first aspect of the present invention provides a method for preparing a controlled-release antibacterial nanofiber hydrogel membrane, the preparation method comprising the following steps:

[0006] (1) mixing the carrier particles with an acetic acid solution, adding EDC / NHS, and activating the mixed solution;

[0007] (2) After the activation is completed, perillaldehyde and a coupling agent are added to carry out a coupling reaction, and a controlled-release antibacterial nanofiber hydrogel membrane is obtained by electrospinning.

[0008] In one embodiment, the preparation method of the carrier particles in step (1) is:

[0009] S1. Zein and 80% ethanol by mass fraction were treated with microwaves at a solid-liquid g / mL ratio of 1:6, with a wave power of 600 W and a microwave time of 10 s to obtain a zein solution, and sodium stearoyl lactylate, water and gelatin were mixed at a mass ratio of 1:20:10 to obtain a gelatin solution;

[0010] S2. Mix the zein solution and gelatin solution in a volume ratio of 1:7-12, put into a supercritical CO2 device, set the temperature to 36.5-39.5°C, maintain the pressure at 6.4-7.3 MPa, maintain the pressure for 30-40 min, and obtain carrier particles.

[0011] In one embodiment, in step (1), the mass ratio of the carrier particles to the acetic acid solution is 12-21:100, the EDC / NHS concentration is 0.5 mM, and the added amount is 3-7% of the volume of the acetic acid solution.

[0012] In one embodiment, the mass ratio of perillaldehyde to carrier particles is 1-8:100.

[0013] In one embodiment, the coupling agent is aminoheptapolyethylene glycol amine, and the coupling agent accounts for 0.3-5% of the mass of the carrier particles.

[0014] In one embodiment, the coupling reaction temperature is 50°C.

[0015] In one embodiment, the voltage of the electrospinning is 19-22 kV.

[0016] A second aspect of the present invention provides a nanofiber hydrogel membrane prepared using the preparation method.

[0017] The third aspect of the present invention provides an application of a nanofiber hydrogel film in food preservation, wherein the food is any one or a combination of livestock, eggs, poultry, aquatic products, dairy products, and vegetable foods.

[0018] In one embodiment, the food is mutton.

[0019] Beneficial effects:

[0020] The present invention uses gelatin and zein as basic raw materials, mixes them, and uses supercritical action to form microporous microspheres, then uses the microspheres as carriers, uses EDC / NHS to activate the surface groups of the carriers, and effectively loads different perillaldehydes, thereby maintaining overall stability and antibacterial properties, while having outstanding insurance effects. The nanofiber hydrogel film provides a new and effective antibacterial packaging for the food industry. DETAILED DESCRIPTION

[0021] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present disclosure belongs. In the event of a conflict, the definitions in this specification shall prevail.

[0022] Unless otherwise indicated, all percentages, parts, ratios, etc. are by weight.

[0023] As used herein, the terms "comprises," "comprising," "having," "having," "containing," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that includes a list of elements is not necessarily limited to only those elements but may also include other elements not expressly listed or inherent to the composition, process, method, article, or apparatus.

[0024] When quantity, weight parts or other numerical value or parameter are given with range, preferred range or a series of upper preferred value and lower preferred value, it should be understood that it specifically discloses all ranges formed by any pair of numerical values ​​of any larger range limit or preferred value and any smaller range limit or preferred value, regardless of whether the range is disclosed respectively. For example, when describing the range of "1 to 5", the described range should be understood to include the range of "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5" etc. Unless otherwise stated, in the description of numerical range herein, the range is intended to include the range end value and all integers, fractions, decimals, etc. within the range.

[0025] In addition, the indefinite articles "a" and "an" before the elements or components of the present disclosure are intended to indicate that the number of occurrences (i.e., occurrences) of the elements or components is not limited. Therefore, "a" or "an" should be understood to include one or at least one, unless it is clearly indicated that the number is singular, otherwise the elements or components in the singular form also include the plural case.

[0026] Unless specifically stated, the materials, methods, and examples described herein are illustrative only and not limiting. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are still described herein.

[0027] The present disclosure is described in detail below.

[0028] Example 1

[0029] The preparation method of the carrier particles is as follows:

[0030] S1. Zein and 80% ethanol by mass fraction were treated with microwaves at a solid-liquid g / mL ratio of 1:6, with a wave power of 600 W and a microwave time of 10 s to obtain a zein solution, and sodium stearoyl lactylate, water and gelatin were mixed at a mass ratio of 1:20:10 to obtain a gelatin solution;

[0031] S2. Mix the zein solution and gelatin solution in a volume ratio of 1:7, put into a supercritical CO2 device, set the temperature to 36.5°C, maintain the pressure at 6.4 MPa, and maintain the pressure for 30 minutes to obtain carrier particles.

[0032] Example 2

[0033] The preparation method of the carrier particles is as follows:

[0034] S1. Zein and 80% ethanol by mass fraction were treated with microwaves at a solid-liquid g / mL ratio of 1:6, with a wave power of 600 W and a microwave time of 10 s to obtain a zein solution, and sodium stearoyl lactylate, water and gelatin were mixed at a mass ratio of 1:20:10 to obtain a gelatin solution;

[0035] S2. Mix the zein solution and gelatin solution in a volume ratio of 1:9, put into a supercritical CO2 device, set the temperature to 37.5°C, maintain the pressure at 6.8 MPa, and maintain the pressure for 35 minutes to obtain carrier particles.

[0036] Example 3

[0037] The preparation method of the carrier particles is as follows:

[0038] S1. Zein and 80% ethanol by mass fraction were treated with microwaves at a solid-liquid g / mL ratio of 1:6, with a wave power of 600 W and a microwave time of 10 s to obtain a zein solution, and sodium stearoyl lactylate, water and gelatin were mixed at a mass ratio of 1:20:10 to obtain a gelatin solution;

[0039] S2. Mix the zein solution and gelatin solution in a volume ratio of 1:12, put into a supercritical CO2 device, set the temperature to 39.5°C, maintain the pressure at 7.3 MPa, and maintain the pressure for 40 minutes to obtain carrier particles.

[0040] Example 4

[0041] The carrier particles used are the carrier particles prepared in Example 2.

[0042] The coupling agent is amino heptapolyethylene glycol amine.

[0043] A method for preparing a controlled-release antibacterial nanofiber hydrogel membrane, characterized in that the preparation method comprises the following steps:

[0044] (1) According to the mass ratio of carrier particles to 70% acetic acid solution of 12:100, the material is taken, the concentration of EDC / NHS is 0.5mM, and the amount added is 3% of the volume of the 70% acetic acid solution of mass fraction, the carrier particles are mixed with the acetic acid solution, and EDC / NHS is added, and the mixed solution is activated at 55°C for 20h;

[0045] (2) According to the mass ratio of perillaldehyde to carrier particles of 1:100 and the coupling agent of 0.3% of the mass of the carrier particles, the material was taken. After the activation was completed, perillaldehyde and the coupling agent were added for coupling reaction. The coupling reaction temperature was 50°C and the reaction time was 3 hours. The electrospinning was performed at a voltage of 19 kV to obtain a controlled-release antibacterial nanofiber hydrogel membrane.

[0046] Example 5

[0047] The carrier particles used are the carrier particles prepared in Example 2.

[0048] The coupling agent is amino heptapolyethylene glycol amine.

[0049] A method for preparing a controlled-release antibacterial nanofiber hydrogel membrane, characterized in that the preparation method comprises the following steps:

[0050] (1) The carrier particles and the acetic acid solution with a mass ratio of 17:100, the EDC / NHS concentration is 0.5 mM, and the amount added is 5% of the volume of the acetic acid solution with a mass fraction of 70%, the carrier particles are mixed with the acetic acid solution, EDC / NHS is added, and the mixed solution is activated at 55° C. for 20 hours;

[0051] (2) According to the mass ratio of perillaldehyde to carrier particles of 5:100, the coupling agent is 3% of the mass of the carrier particles. After the activation is completed, perillaldehyde and the coupling agent are added for coupling reaction. The coupling reaction temperature is 50°C, the reaction time is 3h, and electrospinning is performed at a voltage of 20kV to obtain a controlled-release antibacterial nanofiber hydrogel membrane.

[0052] Example 6

[0053] The carrier particles used are the carrier particles prepared in Example 2.

[0054] The coupling agent is amino heptapolyethylene glycol amine.

[0055] A method for preparing a controlled-release antibacterial nanofiber hydrogel membrane, characterized in that the preparation method comprises the following steps:

[0056] (1) The carrier particles and the acetic acid solution with a mass ratio of 21:100, the EDC / NHS concentration is 0.5 mM, and the amount added is 7% of the volume of the acetic acid solution with a mass fraction of 70%, the carrier particles are mixed with the acetic acid solution, EDC / NHS is added, and the mixed solution is activated at 55° C. for 20 hours;

[0057] (2) According to the mass ratio of perillaldehyde to carrier particles being 8:100 and the coupling agent being 5% of the mass of the carrier particles, after the activation, perillaldehyde and the coupling agent were added for coupling reaction. The coupling reaction temperature was 50°C and the reaction time was 3 h. The electrospinning was performed at a voltage of 22 kV to obtain a controlled-release antibacterial nanofiber hydrogel membrane.

[0058] Comparative Example 1

[0059] It is basically the same as Example 5, except that the carrier particles are made by mixing the zein and gelatin in Example 2 in corresponding proportions.

[0060] Comparative Example 2

[0061] It is basically the same as Example 5, except that the supercritical pressure of the carrier particles in Example 2 is 5.9 MPa.

[0062] Comparative Example 3

[0063] It is basically the same as Example 5, except that the supercritical pressure of the carrier particles in Example 2 is 7.6 MPa.

[0064] The nanofiber hydrogel membranes prepared in Examples 4 to 6 and Comparative Examples 1 to 3 were tested as follows:

[0065] Conductivity of nanofiber hydrogel membrane liquid: The conductivity was measured using a conductivity meter, repeated three times, and the average value was taken.

[0066] The test results are shown in Table 1:

[0067] Table 1

[0068]

[0069] Water vapor transmission rate (WVP) of nanofiber hydrogel membrane: The nanofiber hydrogel membrane was fixed on 20 ml beakers, each containing 10 g of anhydrous calcium carbonate, and placed in an environment of 25°C and 75% humidity. The total weight of the beaker was measured for 7 consecutive days. The WVP values ​​of different nanofiber hydrogel membranes were calculated as follows:

[0070] WVP=(Δm·δ) / (S·T·ΔP)(kg / (m·d·Pa))

[0071] where m is the weight change of the beaker (kg), δ is the thickness of the nanofiber membrane (m), S is the area of ​​the nanofiber hydrogel membrane covering the mouth of the beaker (m2), T is the test time (d), and p is the pressure difference between the inside and outside of the beaker.

[0072] Oxygen permeability (OP) of nanofiber hydrogel membrane: The nanofiber membrane was fixed on 20 mL beakers, each containing 10 g of deoxidizer, and placed in an environment of 25°C and 75% humidity. The total weight of each beaker was measured for 7 consecutive days. The OP calculation formula of the nanofiber hydrogel membrane is as follows:

[0073] OP=(Δm·δ) / (S·T·ΔP)(kgm-1d-1Pa-1)

[0074] Where Δm is the weight change of the beaker (kg), δ is the thickness of the nanofiber membrane (m), S is the area of ​​the nanofiber hydrogel membrane covering the mouth of the beaker (m2), T is the test time (d), and p is the pressure difference between the inside and outside of the beaker.

[0075] The test results are shown in Table 2:

[0076] Table 2

[0077] Example 4 Example 5 Example 6 Comparative Example 1 Comparative Example 2 Comparative Example 3 WVP 2.1 1.5 1.7 2.9 2.7 3.0 OP 1.1 0.5 0.8 4.0 3.1 3.5

[0078] Fresh mutton preservation experiment: Under sterile conditions, mutton was cut into pieces of about 30g and packaged using the nanofiber hydrogel films of Examples 4 to 6 and Comparative Examples 1 to 3, with 5 pieces of mutton in each group. Then, the packaged samples were stored in a refrigerator at 4°C for 0, 2, 4, 6 and 8 days. The total number of colonies was detected, log CFU / g, and the results are shown in Table 3:

[0079] Table 3

[0080] 0 days / TVC 2 days / TVC 4 days / TVC 6 days / TVC 8 days / TVC Example 4 1.43 1.51 2.53 3.62 5.01 Example 5 1.43 1.47 2.34 3.02 4.13 Example 6 1.43 1.50 2.51 3.45 4.89 Comparative Example 1 1.43 2.98 4.55 5.94 7.14 Comparative Example 2 1.43 2.86 3.96 5.21 6.78 Comparative Example 3 1.43 2.88 3.98 5.33 6.88

[0081] Total colony count and pH value analysis: chopped mutton (10 g) was added to 90 mL PBS, and the pH value of refrigerated mutton during storage was analyzed using the determination method of GB 4789.2-2022 standard. The test results are shown in Table 4:

[0082] Table 4

[0083]

[0084]

[0085] Analysis of volatile basic nitrogen (TVB-N) content: 5 g of chopped mutton was added to 25 mL of PBS and then homogenized. After soaking for 30 min, the filtrate should be carefully filtered and collected. According to GB 5009.228-2016 standard, the volatile basic nitrogen content of chopped mutton was determined at different storage periods. The test results are shown in Table 5:

[0086]

[0087] The present invention provides a method for preparing and applying a nanofiber fresh-keeping hydrogel film. There are many methods and ways to implement the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the protection scope of the present invention. All components not specified in this embodiment can be implemented by existing technologies.

Claims

1. A method for preparing a controlled-release antibacterial nanofiber hydrogel membrane, characterized in that: The preparation method comprises the following steps: (1) Mixing the carrier particles with acetic acid solution, adding EDC / NHS, and activating the mixed solution; (2) After activation, perillaldehyde and a coupling agent are added to carry out a coupling reaction, and a controlled-release antibacterial nanofiber hydrogel membrane is obtained by electrospinning.

2. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 1, characterized in that: The preparation method of the carrier particles in step (1) is: S1. Zein and 80% ethanol by mass fraction were treated with microwaves at a solid-liquid g / mL ratio of 1:6, with a wave power of 600 W and a microwave time of 10 s to obtain a zein solution, and sodium stearoyl lactylate, water and gelatin were mixed at a mass ratio of 1:20:10 to obtain a gelatin solution; S2. Mix the zein solution and gelatin solution in a volume ratio of 1:7-12, put into a supercritical CO2 device, set the temperature to 36.5-39.5°C, maintain the pressure at 6.4-7.3 MPa, maintain the pressure for 30-40 min, and obtain carrier particles.

3. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 1, characterized in that: In the step (1), the mass ratio of the carrier particles to the acetic acid solution is 12-21:100, the EDC / NHS concentration is 0.5 mM, and the added amount is 3-7% of the volume of the acetic acid solution.

4. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 1, characterized in that: The mass ratio of the perillaldehyde to the carrier particles is 1 to 8:

100.

5. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 4, characterized in that: The coupling agent is amino heptapolyethylene glycol amine, and the coupling agent accounts for 0.3-5% of the mass of the carrier particles.

6. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 5, characterized in that: The coupling reaction temperature was 50°C.

7. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 6, wherein the voltage of the electrospinning is 19 to 22 kV.

8. A nanofiber hydrogel membrane prepared by the preparation method according to any one of claims 1 to 7.

9. An application of the nanofiber hydrogel film as claimed in claim 8 in food preservation, characterized in that: The food is any one or a combination of livestock, eggs, poultry, aquatic products, dairy products, and vegetable foods.

10. The use according to claim 9, characterized in that: The food is mutton.

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