Ozonized oil microcapsule for inhibiting aspergillus flavus as well as preparation method and application of ozonized oil microcapsule
By preparing ozone oil into microcapsules, the problems of instability and difficulty in storage are solved, the effect of slowly releasing ozone is achieved, effectively inhibiting the growth of Aspergillus aflatoxin and the production of toxins, extending the shelf life of food, and being non-toxic and harmless to the human body and the environment, meeting the requirements of green and environmental protection.
Patent Information
- Application Number
- CN202510223832.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-27
- Publication Date
- 2025-05-27
AI Technical Summary
The prior art has problems of food safety hazards and nutrient damage when inhibiting the growth and production of aspergillus aflatoxin, and the instability of ozone oil and difficulty in storage and transportation.
By preparing ozonated oil into microcapsules, gelatin and gum acacia are used as composite wall materials, and the stability of ozone is improved by CaCl2 curing treatment, and ozone is slowly released under different conditions.
It achieves long-term stability and slow release of ozone oil, effectively inhibits the growth and toxin production of Aspergillus aflatoxin, extends the shelf life of food, and is non-toxic and harmless to the human body and the environment, and meets the requirements of green and environmental protection.
Smart Images
Figure CN120036374A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of functional materials, relates to a video preservative, and specifically relates to an ozonated oil microcapsule capable of inhibiting aflatoxin, and a preparation method and application thereof. Background Art
[0002] Aspergillus flavus is a fungus that can produce aflatoxin, a strong carcinogen. It is a hidden danger that needs to be prevented in foods such as nuts and grains. If nuts and grains are not stored properly, they are easily contaminated by aflatoxin, causing food spoilage and reduced nutritional value. Eating food contaminated with aflatoxin may cause acute poisoning, and long-term low-dose intake will increase the risk of cancers such as liver cancer.
[0003] At present, there are many methods to inhibit the growth of Aspergillus flavus and the production of toxins, such as chemical preservatives, physical sterilization technology, etc. However, the use of chemical preservatives may bring food safety risks, and some consumers are resistant to it; although physical sterilization technology is relatively safe, it may cause certain damage to the quality and nutritional content of food.
[0004] Ozone is a strong oxidant that can destroy the cell structure of Aspergillus flavus, damage the biomacromolecules of Aspergillus flavus and inhibit the synthesis of aflatoxin. Therefore, it has a strong inhibitory effect on the growth of Aspergillus flavus. However, ozone is in gaseous state, unstable and has a difficult odor. Ozonized oil is derived from ozone and has good antibacterial properties, but it is in liquid state, chemically unstable and easy to decompose. It is difficult to maintain the effect of direct use and is inconvenient to store and transport. Summary of the invention
[0005] In view of the shortcomings of the prior art, the purpose of the present invention is to provide an ozonated oil microcapsule for inhibiting Aspergillus flavus and a preparation method and application thereof, so as to improve the stability of the ozonated oil, facilitate storage and transportation, slowly release ozone without obvious odor, be non-toxic and harmless, and effectively inhibit the growth of Aspergillus flavus for a long time.
[0006] In order to achieve the above object, the present invention adopts the following technical solutions:
[0007] A method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus comprises the following steps:
[0008] Step 1, adding (4.5-5) g of ozonated oil to 100 ml of deionized water, and adding Tween-80 according to the volume ratio of ozonated oil to Tween-80 of 3:(1-2), firstly subjecting to high-speed shearing of a homogenizer, and then subjecting to high-pressure homogenization to obtain a stable and uniform ozonated oil emulsion;
[0009] Step 2: Sodium alginate and gelatin are added into deionized water respectively, heated in a water bath at 50-60° C. and stirred until dissolved, to obtain a sodium alginate solution A with a mass fraction of 1-2% and a gelatin solution B with a mass fraction of 3-4%;
[0010] Step 3: Mix the ozonated oil emulsion prepared in step 1 with the sodium alginate solution A and gelatin solution B prepared in step 2 in a volume ratio of 12:5:(5-15), stir until uniform in a 50-60°C water bath, then slowly drop 5% acetic acid under stirring to adjust the pH to 3-5, continue stirring for 30-50 minutes, then stop heating, and quickly cool the reaction system to below 10°C in an ice water bath;
[0011] Step 4: According to the total volume of sodium alginate solution A and gelatin solution B and CaCl 2 The solution volume ratio is (1.6-4): 1. Add CaCl with a mass concentration of 1-3% to the reaction system in step 3. 2 The solution was stirred for 30-50 min, and then centrifuged, washed and dried to obtain ozonated oil microcapsules.
[0012] The present invention also has the following technical features:
[0013] Preferably, the ozonated oil in step 1 is a vegetable oil-based ozonated oil, specifically including any one of olive oil-based ozonated oil, sunflower oil-based ozonated oil and sweet almond oil-based ozonated oil.
[0014] Preferably, the ozone content in the ozonated oil is 100-500 mg / L.
[0015] Preferably, the speed of the homogenizer during the high-speed shearing process in step 1 is (18000-20000) rpm, and the shearing time is 10-30 min.
[0016] Preferably, the rotation speed of the high pressure homogenization in step 1 is 10000 rpm, and the homogenization time is 1 to 3 minutes.
[0017] Preferably, the stirring in step 2, step 3 and step 4 is carried out by using a magnetic stirrer at a rotation speed of 400 to 500 rpm.
[0018] Preferably, the centrifugal washing in step 4 is to add deionized water and centrifuge at 6000 rpm for 3 minutes, which is repeated 3 to 5 times.
[0019] Preferably, the washing in step 4 is washing with deionized water for 3 to 5 times.
[0020] Preferably, the drying in step 4 is freeze drying for 36 to 48 hours.
[0021] The present invention also protects an ozonated oil microcapsule for inhibiting aflatoxin prepared by the method as described above and its application as a food preservative.
[0022] Compared with the prior art, the present invention has the following technical effects:
[0023] The present invention prepares ozonated oil into microcapsules, so that the stability of the ozonated oil is improved. During storage, the ozonated oil microcapsules can slowly release ozone, continuously inhibit the growth of aflatoxin and the production of toxins, and can effectively extend the shelf life of food as a food preservative.
[0024] The present invention uses gelatin and gum arabic as composite wall materials, and uses CaCl 2 The curing treatment makes the microcapsules have good physical and chemical stability. When stored under different temperature and humidity conditions, the morphology and performance of the microcapsules remain basically unchanged, which can effectively prevent the decomposition and leakage of ozonated oil and has good stability.
[0025] The ozonated oil microcapsules prepared by the present invention are easily accepted by users due to their slow ozone release and no obvious ozone smell, and are non-toxic and harmless to human body and environment. No harmful chemical reagents are used in the preparation process, which meets the requirements of green environmental protection and can be widely used in the food field to ensure food safety. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] Figure 1 This is a physical picture of the ozonated oil microcapsule powder prepared in Example 1;
[0027] Figure 2 A photo showing that ozonated oil microcapsule powder is put into a non-woven bag to prepare an antibacterial bag for inhibiting aflatoxin;
[0028] Figure 3 This is a comparison chart of the antibacterial effect of ozonated oil microcapsule antibacterial package on Aspergillus flavus. DETAILED DESCRIPTION
[0029] The specific contents of the present invention are further explained in detail below in conjunction with embodiments.
[0030] The reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. The experimental methods in the following examples where specific experimental conditions are not specified are usually carried out under conventional experimental conditions. Unless otherwise specified, the reagents and raw materials used in the present invention can be obtained commercially.
[0031] The ozonated oil used in the present invention is a commercially available vegetable oil-based ozonated oil, which may be an olive oil-based ozonated oil, a sunflower oil-based ozonated oil or a sweet almond oil-based ozonated oil; wherein the ozone content is 100-500 mg / L.
[0032] Embodiment 1:
[0033] This embodiment provides a method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus, comprising the following steps:
[0034] Step 1: 4.5 g of ozonated oil was added to 100 ml of deionized water, and 3 ml of Tween-80 was added. The mixture was first subjected to high-speed shearing (19000 rpm, 20 min) of a homogenizer, and then subjected to high-pressure homogenization (10000 rpm, 2 min) to obtain a stable and uniform ozonated oil emulsion.
[0035] Step 2: Take 0.22g sodium alginate and 0.68g gelatin, add them into 20ml deionized water respectively, heat in a 50℃ water bath and stir magnetically at 450rpm until dissolved, to obtain sodium alginate solution A and gelatin solution B;
[0036] Step 3: 48 ml of ozonated oil emulsion, 20 ml of solution A and 20 ml of solution B were fully mixed under magnetic stirring (50°C, 450 rpm) and set aside; 5% acetic acid was slowly added dropwise while maintaining the stirring speed to adjust the pH to 3.5, and then stirring was continued for 30 min; heating was stopped, and the reaction system was quickly cooled to below 10°C in an ice water bath;
[0037] Step 4: Add 20 ml of 2.5% CaCl 2 The solution solidified the microcapsules, and stirring was continued for 30 minutes after solidification; the microcapsules were washed three times by centrifugation at 6000 rpm for 3 minutes to obtain precipitates, i.e., wet microcapsules, which were freeze-dried for 48 hours to obtain dry ozonated oil microcapsules.
[0038] Embodiment 2:
[0039] This embodiment provides a method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus, comprising the following steps:
[0040] Step 1: 4.5 g of ozonated oil was added to 100 ml of deionized water, and 3 ml of Tween-80 was added. The mixture was first subjected to high-speed shearing (18000 rpm, 30 min) of a homogenizer, and then subjected to high-pressure homogenization (10000 rpm, 1 min) to obtain a stable and uniform ozonated oil emulsion.
[0041] Step 2: Take 0.22g sodium alginate and 1.25g gelatin, add them into 20ml and 40ml deionized water respectively, heat in a 55℃ water bath and stir at 450rpm until dissolved, to obtain sodium alginate solution A and gelatin solution B;
[0042] Step 3: 48 ml of ozonated oil emulsion, 20 ml of sodium alginate solution A and 40 ml of gelatin solution B were fully mixed under magnetic stirring (55°C, 450 rpm) and set aside; 5% acetic acid was slowly added dropwise to adjust the pH to 4 while maintaining the stirring speed, and then stirring was continued for 50 min; heating was stopped, and the reaction system was quickly cooled to below 10°C in an ice water bath;
[0043] Step 4: Add 25 ml of 3% CaCl 2 The solution solidified the microcapsules, and stirring was continued for 50 minutes after solidification; the microcapsules were washed by centrifugation at 6000 rpm for 3 minutes for 4 times to obtain precipitates, i.e., wet microcapsules, which were freeze-dried for 36 hours to obtain dry ozonated oil microcapsules.
[0044] Embodiment 3:
[0045] This embodiment provides a method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus, comprising the following steps:
[0046] Step 1: 6 g of ozonated oil was added to 100 ml of deionized water, and 3.3 ml of Tween-80 was added, and the mixture was first subjected to high-speed shearing (20,000 rpm, 10 min) of a homogenizer, and then subjected to high-pressure homogenization (10,000 rpm, 3 min) to obtain a stable and uniform ozonated oil emulsion;
[0047] Step 2: Take 0.44 g of sodium alginate and 2.5 g of gelatin, add them into 20 ml and 60 ml of deionized water respectively, heat in a 60°C water bath and stir at 400 rpm until dissolved, to obtain sodium alginate solution A and gelatin solution B;
[0048] Step 3: 48 ml of ozonated oil emulsion, 20 ml of sodium alginate solution A and 60 ml of gelatin solution B were fully mixed under magnetic stirring (60° C., 400 rpm) and set aside; 5% acetic acid was slowly added dropwise while maintaining the stirring speed to adjust the pH to 3, and then stirring was continued for 40 min; heating was stopped, and the reaction system was quickly cooled to below 10° C. in an ice water bath;
[0049] Step 4: Add 20 ml of 1% CaCl 2 The solution solidifies the microcapsules, and stirring is continued for 40 minutes after solidification; the microcapsules are washed by centrifugation for 5 times at 6000 rpm for 3 minutes to obtain precipitates, i.e., wet microcapsules; and the microcapsules are freeze-dried for 40 hours to obtain dry ozonated oil microcapsules.
[0050] Embodiment 4:
[0051] This embodiment provides a method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus, comprising the following steps:
[0052] Step 1: 4.5 g of ozonated oil was added to 100 ml of deionized water, and 1.54 ml of Tween-80 was added, and the mixture was first subjected to high-speed shearing (19000 rpm, 20 min) of a homogenizer, and then subjected to high-pressure homogenization (10000 rpm, 2 min) to obtain a stable and uniform ozonated oil emulsion;
[0053] Step 2: Take 0.3 g of sodium alginate and 0.8 g of gelatin, add them into 20 ml of deionized water respectively, heat in a 40°C water bath and stir at 500 rpm until dissolved, to obtain sodium alginate solution A and gelatin solution B;
[0054] Step 3: 48 ml of ozonated oil emulsion, 20 ml of sodium alginate solution A and 20 ml of gelatin solution B were fully mixed under magnetic stirring (55°C, 500 rpm) and set aside; 5% acetic acid was slowly added dropwise while maintaining the stirring speed to adjust the pH to 5, and then stirring was continued for 30 min; heating was stopped, and the reaction system was quickly cooled to below 10°C in an ice water bath;
[0055] Step 4: Add 25 ml of 2% CaCl 2 The solution solidifies the microcapsules, and stirring is continued for 50 minutes after solidification; the microcapsules are washed three times by centrifugation at 6000 rpm for 3 minutes to obtain precipitates, i.e., wet microcapsules; and the microcapsules are freeze-dried for 48 hours to obtain dry ozonated oil microcapsules.
[0056] Figure 1 This is a physical picture of the ozonated oil microcapsule powder prepared in Example 1;
[0057] Figure 2 A photo showing that ozonated oil microcapsule powder is put into a non-woven bag to prepare an antibacterial bag for inhibiting aflatoxin;
[0058] The antibacterial bag prepared by using the ozonated oil microcapsules prepared in Example 1 was tested for its low inhibition on the growth of Aspergillus flavus. The specific steps are as follows:
[0059] 1. Preparation of bacterial suspension: Inoculate the Aspergillus flavus strain into liquid culture medium and culture it under appropriate conditions. After the culture is completed, collect the bacteria by centrifugation.
[0060] 2. Inoculation of Aspergillus flavus: In the clean bench, use a pipette to absorb 3 mL of 105 / mL bacterial suspension and evenly spread it on the surface of the solidified PDA culture medium to make the Aspergillus flavus evenly distributed in the culture dish;
[0061] 3. Place ozonated oil microcapsules: Use sterile tweezers to place the non-woven bag without ozonated oil microcapsules, the non-woven bag antibacterial bag containing 300 mg of ozonated oil microcapsules, and the non-woven bag antibacterial bag containing 600 mg of ozonated oil microcapsules in the culture dish coated with Aspergillus flavus;
[0062] 4. Cultivation and observation: Place the culture dish in a constant temperature incubator and culture it at a temperature of 37°C and a humidity of 95%. Observe the growth of Aspergillus flavus after one week.
[0063] Figure 3 This is a comparison chart of the antibacterial effect of ozonated oil microcapsule antibacterial package on Aspergillus flavus. Figure 3 As shown in the figure, the growth of Aspergillus flavus in the culture dish where the non-woven bag antibacterial bag containing 600 mg of ozonated oil microcapsules was placed was significantly weakened compared with the non-woven bag antibacterial bag without ozonated oil microcapsules and the non-woven bag antibacterial bag containing 300 mg of ozonated oil microcapsules, proving that ozonated oil microcapsules have the effect of inhibiting the growth of Aspergillus flavus.
[0064] The ozonated oil microcapsules prepared by the present invention are easily accepted by users due to their slow ozone release and no obvious ozone smell, and are non-toxic and harmless to human body and environment. No harmful chemical reagents are used in the preparation process, which meets the requirements of green environmental protection and can be widely used in the food field to ensure food safety.
[0065] It should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it; without departing from the concept of the present invention, the deduction or replacement made by those skilled in the art shall fall within the protection scope of the present invention.
Claims
1. A method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus, characterized in that: The following steps are involved: Step 1, adding (4.5-5) g of ozonated oil to 100 ml of deionized water, and adding Tween-80 according to the volume ratio of ozonated oil to Tween-80 of 3:(1-2), firstly subjecting to high-speed shearing of a homogenizer, and then subjecting to high-pressure homogenization to obtain a stable and uniform ozonated oil emulsion; Step 2: Sodium alginate and gelatin are added into deionized water respectively, heated in a water bath at 50-60° C. and stirred until dissolved, to obtain a sodium alginate solution A with a mass fraction of 1-2% and a gelatin solution B with a mass fraction of 3-4%; Step 3: Mix the ozonated oil emulsion prepared in step 1 with the sodium alginate solution A and gelatin solution B prepared in step 2 in a volume ratio of 12:5:(5-15), stir until uniform in a 50-60°C water bath, then slowly drop 5% acetic acid under stirring to adjust the pH to 3-5, continue stirring for 30-50 minutes, then stop heating, and quickly cool the reaction system to below 10°C in an ice water bath; Step 4: Add 1-3% CaCl2 solution to the reaction system of step 3 at a volume ratio of (1.6-4):1 of the total volume of sodium alginate solution A and gelatin solution B to the volume of CaCl2 solution and continue stirring for 30-50 minutes, then centrifuge, wash and dry to obtain ozonated oil microcapsules.
2. The method for preparing the ozonated oil microcapsules for inhibiting Aspergillus flavus according to claim 1, characterized in that: The ozonated oil described in step 1 is a vegetable oil-based ozonated oil, specifically including any one of olive oil-based ozonated oil, sunflower oil-based ozonated oil and sweet almond oil-based ozonated oil.
3. The method for preparing the ozonated oil microcapsules for inhibiting Aspergillus flavus according to claim 1, characterized in that: The ozone content in the ozonated oil is 100-500 mg / L.
4. The method for preparing the ozonated oil microcapsules for inhibiting Aspergillus flavus according to claim 1, characterized in that: The speed of the homogenizer during the high-speed shearing process in step 1 is (18000-20000) rpm, and the shearing time is 10-30 min.
5. The method for preparing the ozonated oil microcapsules for inhibiting Aspergillus flavus according to claim 1, characterized in that: The rotation speed of the high pressure homogenizer described in step 1 is 10000 rpm, and the homogenization time is 1 to 3 minutes.
6. The method for preparing the ozonated oil microcapsules for inhibiting Aspergillus flavus according to claim 1, characterized in that: The stirring in step 2, step 3 and step 4 is carried out by using a magnetic stirrer at a rotation speed of 400 to 500 rpm.
7. The method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus according to claim 1, characterized in that: The centrifugal washing described in step 4 is to add deionized water and centrifuge at 6000 rpm for 3 minutes, and repeat 3 to 5 times.
8. The method for preparing ozonated oil microcapsules for inhibiting Aspergillus flavus according to claim 1, characterized in that: The drying described in step 4 is freeze drying for 36 to 48 hours.
9. Ozoned oil microcapsules for inhibiting aflatoxin prepared by the method according to any one of claims 1 to 8.
10. Use of the ozonated oil microcapsules for inhibiting aflatoxin as claimed in claim 9 as a food preservative.