Preparation method and application of a controlled-release bacteriostatic nanofiber hydrogel film

Abstract

The application belongs to the technical field of food processing, and particularly relates to a preparation method of a controlled-release antibacterial nanofiber hydrogel film and application thereof. The nanofiber hydrogel film is prepared by using gelatin and corn alcohol-soluble protein as basic raw materials, mixing, forming microporous microspheres by using supercritical action, taking the microspheres as carriers, and activating surface groups of the carriers by EDC / NHS to effectively load different perillaldehydes, so that the overall stability and antibacterial performance are maintained, and the effect is outstanding. The nanofiber hydrogel film provides a new and effective antibacterial packaging for the food industry.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology, specifically relating to a method for preparing a controlled-release antibacterial nanofiber hydrogel membrane and its application. Background Technology

[0002] Lamb is rich in nutrients and is loved by people all over the world for its unique flavor and high protein content. Currently, the market mainly offers chilled, frozen, and fresh lamb. Chilled lamb, with its more tender texture, is gradually becoming the mainstream product. However, chilled lamb is susceptible to microbial contamination and spoilage during production and transportation. Therefore, the main challenges are extending the shelf life of chilled lamb and improving product quality, making the search for a new and effective preservation technology imperative.

[0003] Zeatin, chitosan, sodium alginate, starch, and methylcellulose are commonly used carrier matrices for preparing edible films. Active films made from single biopolymers, due to their hydrophilic and easily decomposed properties, cannot meet the actual needs of food packaging and preservation. This deficiency is often compensated for by combining different biopolymers to form highly efficient water vapor barrier and hydrophobic properties. It has been reported that chitosan or zeatin, through enhanced interactions such as electrostatics and hydrogen bonds between functional groups, can prevent the rapid decomposition of gelatin-based polymer films upon contact with water. However, these films cannot continuously inhibit microbial growth during meat preservation and storage, leading to a "burst release" problem of antibacterial substances in the film. Since essential oils are volatile substances, current research has failed to effectively control the release of essential oil-based active substances, resulting in weak controlled release of antibacterial active substances, low antibacterial efficiency, and poor barrier performance in bio-based packaging films. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides a method for preparing a nanofiber hydrogel membrane that can effectively load perillaldehyde and effectively release controlled-release antibacterial agents. Simultaneously, the nanofiber hydrogel membrane exhibits significant antibacterial properties and stable preservation effects, significantly extending the shelf life of fresh mutton and making it suitable for packaging and preserving fresh meats such as mutton.

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

[0006] (1) Mix the carrier particles with acetic acid solution, add EDC / NHS, and activate the mixed solution;

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

[0008] In one embodiment, the method for preparing the carrier particles in step (1) is as follows:

[0009] S1. Mix zein and 80% ethanol at a solid-liquid g / mL ratio of 1:6, microwave at a power of 600W for 10s to obtain a zein solution. Mix sodium stearoyl lactylate, water and gelatin 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, place them in a supercritical CO2 device, set the temperature to 36.5-39.5℃, maintain the pressure at 6.4-7.3MPa, and maintain the pressure for 30-40 minutes to 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 amount added 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 to 8:100.

[0013] In one embodiment, the coupling agent is aminoheptapolyethylene glycolamine, and the coupling agent accounts for 0.3 to 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 a preparation method.

[0017] The third aspect of this invention provides an application of a nanofiber hydrogel membrane in food preservation, wherein the food is any one or a combination of several of the following: livestock, eggs, poultry, aquatic products, dairy products, and vegetables.

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

[0019] Beneficial effects:

[0020] This invention uses gelatin and zein as basic raw materials. Through mixing and supercritical fluid extraction, microporous microspheres are formed. These microspheres are then used as carriers, and EDC / NHS is used to activate the surface groups of the carriers, effectively loading different perillaldehydes. This maintains the overall stability and antibacterial properties, while also providing excellent preservation effects. This nanofiber hydrogel membrane provides a novel and effective antibacterial packaging for the food industry. Detailed Implementation

[0021] Unless otherwise specified, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In case of any discrepancy, the definitions in this specification shall prevail.

[0022] Unless otherwise stated, all percentages, portions, proportions, etc. are by weight.

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

[0024] When quantities, parts by weight, or other numerical values ​​or parameters are given as ranges, preferred ranges, or a series of upper and lower preferred values, it should be understood that they specifically disclose all ranges formed by any pair of values ​​of any larger or preferred range limit and any smaller or preferred range limit, regardless of whether the ranges are disclosed separately. For example, when describing a range of "1 to 5", the described range should be understood to include ranges such as "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, where numerical ranges are described herein, the range is intended to include the range endpoints as well as all integers, fractions, decimals, etc., within that range.

[0025] Furthermore, the indefinite articles “a” and “an” preceding the elements or components of this disclosure are intended to indicate that there is no limitation on the number of times the said element or component appears (i.e., occurs). Therefore, “a” or “an” should be understood to include one or at least one, and unless the quantity is explicitly stated to be singular, the singular form of the said element or component also includes the plural case.

[0026] Unless otherwise specified, the materials, methods, and examples described herein are exemplary and not limiting. While similar or equivalent methods and materials may be used in implementing or testing this disclosure, suitable methods and materials are also described herein.

[0027] This disclosure is described in detail below.

[0028] Example 1

[0029] The method for preparing the carrier particles is as follows:

[0030] S1. Mix zein and 80% ethanol at a solid-liquid g / mL ratio of 1:6, microwave at a power of 600W for 10s to obtain a zein solution. Mix sodium stearoyl lactylate, water and gelatin at a mass ratio of 1:20:10 to obtain a gelatin solution.

[0031] S2. Mix the corn gliadin solution and gelatin solution in a volume ratio of 1:7, place them in a supercritical CO2 device, set the temperature to 36.5℃, maintain the pressure at 6.4MPa, and maintain the pressure for 30 minutes to obtain carrier particles.

[0032] Example 2

[0033] The method for preparing the carrier particles is as follows:

[0034] S1. Mix zein and 80% ethanol at a solid-liquid g / mL ratio of 1:6, microwave at a power of 600W for 10s to obtain a zein solution. Mix sodium stearoyl lactylate, water and gelatin at a mass ratio of 1:20:10 to obtain a gelatin solution.

[0035] S2. Mix the corn gliadin solution and gelatin solution in a volume ratio of 1:9, place them in a supercritical CO2 device, set the temperature to 37.5℃, maintain the pressure at 6.8MPa, and maintain the pressure for 35 minutes to obtain carrier particles.

[0036] Example 3

[0037] The method for preparing the carrier particles is as follows:

[0038] S1. Mix zein and 80% ethanol at a solid-liquid g / mL ratio of 1:6, microwave at a power of 600W for 10s to obtain a zein solution. Mix sodium stearoyl lactylate, water and gelatin at a mass ratio of 1:20:10 to obtain a gelatin solution.

[0039] S2. Mix the corn gliadin solution and gelatin solution at a volume ratio of 1:12, place them in a supercritical CO2 device, set the temperature to 39.5℃, maintain the pressure at 7.3MPa, and maintain the pressure for 40 minutes to obtain carrier particles.

[0040] Example 4

[0041] The carrier particles used were those prepared in Example 2.

[0042] The coupling agent is aminoheptapolyethylene glycolamine.

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

[0044] (1) Take materials according to the mass ratio of carrier particles to 70% acetic acid solution of 12:100, EDC / NHS concentration of 0.5mM, the amount added is 3% of the volume of 70% acetic acid solution, mix the carrier particles with acetic acid solution, add EDC / NHS, and activate the mixed solution at 55℃ for 20h.

[0045] (2) The materials were taken according to the mass ratio of perillaldehyde to carrier particles of 1:100 and the coupling agent of 0.3% of the mass of carrier particles. After activation, perillaldehyde and coupling agent were added to carry out coupling reaction. The coupling reaction temperature was 50℃ and the reaction time was 3h. Electrospinning was carried out at a voltage of 19kV to obtain a controlled-release antibacterial nanofiber hydrogel membrane.

[0046] Example 5

[0047] The carrier particles used were those prepared in Example 2.

[0048] The coupling agent is aminoheptapolyethylene glycolamine.

[0049] A method for preparing a controlled-release antibacterial nanofiber hydrogel membrane, characterized in that the preparation method includes 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 0.5mM, and the amount added is 5% of the volume of the 70% acetic acid solution. The carrier particles and the acetic acid solution are mixed, and EDC / NHS is added. The mixed solution is activated at 55°C for 20h.

[0051] (2) Perillaldehyde and carrier particles were added at a mass ratio of 5:100, and the coupling agent was 3% of the mass of the carrier particles. After activation, perillaldehyde and coupling agent were added to carry out the coupling reaction. The coupling reaction temperature was 50℃ and the reaction time was 3h. Electrospinning was performed at a voltage of 20kV to obtain a controlled-release antibacterial nanofiber hydrogel membrane.

[0052] Example 6

[0053] The carrier particles used were those prepared in Example 2.

[0054] The coupling agent is aminoheptapolyethylene glycolamine.

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

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

[0057] (2) According to the mass ratio of perillaldehyde to carrier particles of 8:100, the coupling agent is 5% of the mass of carrier particles. After activation, perillaldehyde and coupling agent are added to carry out coupling reaction. The coupling reaction temperature is 50℃ and the reaction time is 3h. Electrospinning is performed at a voltage of 22kV 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 zein and gelatin in the corresponding proportions as in Example 2.

[0060] Comparative Example 2

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

[0062] Comparative Example 3

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

[0064] The nanofiber hydrogel membranes prepared in Examples 4-6 and Comparative Examples 1-3 were subjected to the following tests:

[0065] The conductivity of the nanofiber hydrogel membrane was measured using a conductivity meter. The measurement was 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 membranes: Nanofiber hydrogel membranes were 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 beakers 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 change in beaker weight (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 membranes: Nanofiber membranes were fixed on 20mL beakers, each containing 10g of oxygen absorber, and placed in an environment of 25℃ and 75% humidity. The total weight of each beaker was measured for 7 consecutive days. The OP of the nanofiber hydrogel membrane is calculated using the following formula:

[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 inside and outside the beaker.

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

[0076] Table 2

[0077] 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 aseptic conditions, mutton was cut into pieces of approximately 30g each and packaged using nanofiber hydrogel membranes (Examples 4-6, Comparative Examples 1-3), with 5 pieces of mutton wrapped in each package. The packaged samples were then stored in a refrigerator at 4°C for 0, 2, 4, 6, and 8 days. The total bacterial count (log CFU / g) was measured, and the results are shown in Table 3.

[0079] Table 3

[0080] 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 bacterial count and pH analysis: 10g of chopped mutton was added to 90mL of PBS, and the pH value of the refrigerated mutton during storage was analyzed using the determination method of GB 4789.2-2022. The test results are shown in Table 4.

[0082] Table 4

[0083]

[0084]

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

[0086]

[0087] This invention provides a method for preparing and applying a nanofiber preservation hydrogel film. Many methods and approaches exist for implementing this technical solution; the above description is merely a preferred embodiment of the invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this invention, and these improvements and modifications should also be considered within the scope of protection of this invention. All components not explicitly stated in this embodiment can be implemented using existing technologies.

Claims

1. A method for preparing a controlled-release antibacterial nanofiber hydrogel membrane, characterized in that, The preparation method includes the following steps: (1) Mix the carrier particles with acetic acid solution, add EDC / NHS, and activate the mixed solution; (2) After activation, perillaldehyde and coupling agent are added to carry out coupling reaction, and controlled-release antibacterial nanofiber hydrogel membrane is obtained by electrospinning. The method for preparing the carrier particles in step (1) is as follows: S1. Mix zein and 80% ethanol at a solid-liquid g / mL ratio of 1:6, microwave at a power of 600W for 10s to obtain a zein solution. Mix sodium stearoyl lactylate, water and gelatin 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, place them in a supercritical CO2 device, set the temperature to 36.5-39.5℃, maintain the pressure at 6.4-3MPa, and maintain the pressure for 30-40 minutes to obtain carrier particles; The coupling agent is aminoheptapolyethylene glycolamine, and the coupling agent accounts for 0.3 to 5% of the mass of the carrier particles.

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

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

100.

4. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 1, characterized in that, The coupling reaction temperature is 50°C.

5. The method for preparing the controlled-release antibacterial nanofiber hydrogel membrane according to claim 4, wherein the voltage of electrospinning is 19-22 kV.

6. A nanofiber hydrogel membrane prepared using the preparation method according to any one of claims 1 to 5.

7. The application of the nanofiber hydrogel membrane as described in claim 6 in food preservation, characterized in that, The food is any one or a combination of several of the following: livestock, eggs, poultry, aquatic products, dairy products, and vegetables.

8. The application according to claim 7, characterized in that, The food in question is mutton.

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