Copper-zinc chelated composite material, preparation method and application of copper-zinc chelated composite material in feed

Abstract

The invention relates to the technical field of composite materials, and discloses a copper-zinc chelated composite material, a preparation method thereof and an application of the copper-zinc chelated composite material in a feed. The preparation method comprises the following steps: modifying rice bran meal by adopting physical and chemical methods to obtain modified rice bran meal; further modifying the composition of the rice bran meal by adopting a biological enzymolysis method to obtain functionalized rice bran meal; reacting copper sulfate and zinc sulfate with glycine and calcium carbonate, ending the reaction when no bubbles are generated in the system, and centrifuging to obtain a copper ion solution and a zinc ion solution; and mixing the functionalized rice bran meal with a copper ion solution and a zinc ion solution, adding a lactic acid bacteria solution for fermentation, and after the reaction is finished, obtaining the efficient copper-zinc chelating composite material. The high-efficiency copper-zinc chelated composite material can be used as a trace element supplement in a feed.

Description

Technical Field

[0001] This invention relates to the field of composite material technology, specifically to a method for preparing a high-efficiency copper-zinc chelate composite material and its application in feed. Background Technology

[0002] Copper and zinc are essential trace elements in animals, participating in numerous biochemical reactions. Copper is present in almost every tissue of the body, participating in biochemical reactions such as acting as a catalyst for redox reactions, participating in neurotransmission and immune system function, maintaining bone and connective tissue health, and participating in pigment synthesis. Zinc has multiple functions in the body, including promoting enzyme activity, participating in DNA and protein synthesis, maintaining normal immune system function, and promoting cell differentiation and growth. Furthermore, copper and zinc are important components of copper-zinc superoxide dismutase, which helps reduce free radical damage and maintain cell stability.

[0003] For a long time, the form of adding copper and zinc trace elements to feed has mainly been inorganic salts such as zinc sulfate, zinc chloride, copper sulfate, and basic copper chloride. Because inorganic trace elements have low absorption rates in animals, they are often supplemented in excessive amounts. This leads to a series of problems, including high dosage, strong irritation, high acid-binding capacity, large emissions, soil pollution, and resource scarcity. Therefore, finding trace element forms with higher absorption rates, reducing the amount of trace elements added to feed, and ensuring animal health is a problem facing researchers.

[0004] Rice bran meal is the product of rice bran after oil extraction during the processing of rice into polished rice. my country's annual rice bran meal production exceeds 3 million tons. Rice bran protein has low allergenicity and a balanced amino acid composition, making it a potential nutritional resource. Enzymatic hydrolysis can produce physiologically active rice bran peptides, such as antioxidant peptides, opioid antagonist peptides, and angiotensin-converting enzyme (ACE) inhibitory peptides, but metal chelate peptides are rarely reported. Rice bran polysaccharides are important functional components of rice bran meal, a class of complex heteropolymers with various effects including antioxidant activity, improved immunity, hypoglycemia, hypolipidemia, antibacterial activity, anti-radiation properties, and preservation. Existing technologies, such as Chinese patent application CN2021104814437, disclose a method for preparing and applying a zinc chelate of rice bran polysaccharide. By extracting rice bran polysaccharide, a high-content zinc chelate of polysaccharide is obtained, exhibiting significant antioxidant effects. However, this additive is only a single chelating ligand, and its absorption channels in the body are limited. If a composite material can be developed to achieve multiple chelation of various metal ions and realize target release and multi-channel absorption, it will be of great significance to the effective application of supplements. Summary of the Invention

[0005] The purpose of this invention is to achieve high-value utilization of biomass resources by applying composite material design concepts and chemical chelation technology to provide a method for preparing a high-efficiency copper-zinc chelate composite material and its application in feed. This high-efficiency copper-zinc chelate composite material has excellent synergistic performance. Rice bran meal alone has limited adsorption capacity, while copper and zinc inorganic salts alone have low bioavailability and strong irritation. By combining the two through the technical means of this invention, a brand-new material is produced. Its performance (chelation stability, bioavailability, and safety) is far superior to the simple sum of the performance of each individual component, and it can be used in feed as a trace element supplement.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] A method for preparing a high-efficiency copper-zinc chelate composite material includes the following steps:

[0008] Step (1): Crush the rice bran meal through a 20-60 mesh sieve, and take the powder that passes through the sieve for steam explosion treatment;

[0009] Step (2): Add 1.0-1.5 mol / L hydrochloric acid to the steam-exploded rice bran meal for acidification treatment to obtain modified rice bran meal;

[0010] Step (3): Mix the modified rice bran meal with phytase, xylanase, cellulase, glucoamylase and acidic protease until uniform, and react. After the reaction is completed, dry at 65-95℃ for 6-24 hours to obtain functionalized rice bran meal.

[0011] Step (4): Mix copper sulfate, zinc sulfate, glycine, calcium carbonate and water, stir and react. After the reaction is complete, centrifuge at 3000-8000 r / min for 5-15 min, collect the supernatant to obtain a copper ion + zinc ion solution.

[0012] Step (5): Mix the functionalized rice bran meal with copper ion + zinc ion solution evenly, add lactic acid bacteria liquid, ferment for 12-72h, after the reaction is completed, dry at 50-65℃ for 4-24h to obtain a high-efficiency copper-zinc chelate composite material.

[0013] Preferably, in step (1): the rice bran meal is pulverized through a 20-60 mesh sieve, and the reaction conditions for steam explosion treatment are: steam pressure 0.5-1.2 MPa, temperature 120-180℃, pressure holding time 1-5 min, venting (explosion) time 150-350 ms, and the moisture content of the rice bran meal during steam explosion treatment is 15-30%; preferably, the steam pressure is 0.6-1.0 MPa (more preferably 0.7-0.9 MPa), temperature 130-170℃ (more preferably 140-160℃), pressure holding time 2-4 min (more preferably 2.5-3.5 min), venting (explosion) time 180-320 ms (more preferably 220-280 ms), and the moisture content of the rice bran meal is 20-25% (more preferably 21.5-23.5%).

[0014] Preferably, in step (2): during the acidification treatment, 1.1-1.4 mol / L (more preferably 1.2-1.3 mol / L) hydrochloric acid is added, and the mass ratio of rice bran meal after steam explosion treatment to hydrochloric acid is 4:3-8, preferably 4:4-7 (more preferably 4:5-6); the reaction conditions are: reacting at room temperature for 30-60 min; preferably reacting at room temperature for 40-50 min (more preferably 42-48 min).

[0015] Preferably, in step (3): the amount of phytase added is 100-250 U / g modified rice bran meal, the amount of xylanase added is 10-800 U / g modified rice bran meal, the amount of cellulase added is 1500-3500 U / g modified rice bran meal, the amount of saccharifying enzyme added is 50-200 U / g modified rice bran meal, and the amount of acidic protease added is 250-2000 U / g modified rice bran meal. Preferably, the amount of phytase added is 150-200 U / g modified rice bran meal (more preferably 160-180 U / g), the amount of xylanase added is 50-500 U / g modified rice bran meal (more preferably 100-300 U / g), the amount of cellulase added is 2200-2800 U / g modified rice bran meal (more preferably 2400-2600 U / g), and the amount of saccharifying enzyme added is 100-150 U / g modified rice bran meal (more preferably 120-140 U / g). The amount of acidic protease added is 500-1000 U / g of modified rice bran (more preferably 600-800 U / g); the reaction conditions are: reacting at 40-60℃ for 3-7h, preferably at 45-55℃ (more preferably 47-53℃) for 4-6h (more preferably 4.5-5.5h); after the reaction, drying is preferably at 75-85℃ (more preferably 78-82℃), and the drying time is preferably 8-20h (more preferably 10-16h).

[0016] Preferably, in step (4), the mass ratio of copper ions, zinc ions, glycine, calcium carbonate, and water is 1:2-15:6-15:4-10:50-200, and more preferably, the mass ratio of copper ions, zinc ions, glycine, calcium carbonate, and water is 1:4-12:8-13:5-9:80-150 (more preferably 1:5-10:9-12:6-8:100-120); the reaction conditions are 30-60℃, preferably 40-50℃ (more preferably 42-48℃); after the reaction is completed, centrifuge at a speed of 4000-7000 r / min (more preferably 5000-6000 r / min) and a centrifugation time of 6-12 min (more preferably 8-10 min), and collect the supernatant.

[0017] Preferably, in step (4): the reaction ends when no more bubbles are generated in the solution, and the time is usually 0.5-2.5h (preferably 1-2h).

[0018] Preferably, in step (4): copper sulfate is anhydrous copper sulfate and / or copper sulfate pentahydrate, and zinc sulfate is zinc sulfate monohydrate and / or zinc sulfate heptahydrate.

[0019] Preferably, in step (5): the mass ratio of functionalized rice bran meal, copper ion + zinc ion solution, and lactic acid bacteria liquid is 20:15-30:2-6, preferably 20:20-25:3-5 (more preferably 20:21-24:3.5-4.5); the pH value of the lactic acid bacteria liquid is 3.5-5.0, preferably 4.0-4.5 (more preferably 4.1-4.4); the fermentation temperature is 25-40℃, preferably 30-37℃ (more preferably 32-35℃); the fermentation time is 12-72h, preferably 24-48h (more preferably 28-42h); after the reaction is completed, it is preferably dried at 55-65℃ (more preferably 57-63℃), and the drying time is preferably 6-20h (more preferably 8-12h) to obtain a high-efficiency copper-zinc chelate composite material.

[0020] Preferably, in step (5), the fermentation strain of the lactic acid bacteria liquid is a commonly used lactic acid bacteria in production, including but not limited to one or more of Lactobacillus, Streptococcus, Enterococcus, and Pediococcus; preferably, the fermentation strain of the lactic acid bacteria liquid is Pediococcus, such as Pyrococcus lactis.

[0021] The high-efficiency copper-zinc chelate composite material described above can be used in animal feed, either directly as animal feed or as an animal feed additive.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0023] 1. Rice bran meal alone has limited adsorption capacity. This patent modifies and functionalizes rice bran meal to completely change its composition, making it rich in disaccharides, oligosaccharides, polysaccharides, and small peptides, which can efficiently chelate copper and zinc ions. Pure copper and zinc inorganic salts have low bioavailability and strong irritation. This patent produces a brand-new material through metal ion activation reaction, microbial fermentation and other technical means. Its performance (chelation stability, bioavailability and safety) is far superior to the simple sum of the performance of each single component.

[0024] 2. An indicator for the metal ion activation reaction system was designed to make the activation reaction of metal ions intuitive and observable.

[0025] 3. The composite material prepared by this invention achieves multiple chelation of various metal ions, enabling target release and multi-channel absorption, thereby improving the absorption and utilization rate of copper and zinc trace elements and achieving a highly efficient and safe effect. Attached Figure Description

[0026] Figure 1 The graph shows the determination of polysaccharide and protein indicators of rice bran meal before and after modification at room temperature for 1 hour.

[0027] Figure 2 The graph shows the determination of polysaccharide and protein indicators of rice bran meal before and after modification under heating conditions for 1 hour.

[0028] Figure 3 The graph shows the determination of polysaccharide and protein indicators of rice bran meal before and after modification under heating conditions for 2 hours.

[0029] Figure 4 The graph shows the determination of polysaccharide indexes of rice bran meal before and after functionalization treatment in each embodiment.

[0030] Figure 5 The graph shows the determination of protein peptide indicators before and after functionalization treatment of rice bran meal in each embodiment. Detailed Implementation

[0031] The following examples are intended to illustrate the invention and not to further limit it.

[0032] In the examples and comparative examples: phytase was from Inner Mongolia Yiduoli Biotechnology Co., Ltd., model Yiduolin 10000, enzyme activity ≥10000U / g; xylanase was from Henan Ferment Doctor Bioengineering Co., Ltd., model III, enzyme activity ≥100000U / g; cellulase was from Ningxia Xiasheng Industrial Group Co., Ltd., model H, enzyme activity ≥100000U / g; glucoamylase was from Shandong Longkete Enzyme Preparation Co., Ltd., food grade, enzyme activity ≥100000U / g; acidic protease was from Henan Wanbang Chemical Technology Co., Ltd., trade name Yuwanbang, enzyme activity ≥50000U / g.

[0033] The lactic acid bacteria used were Pediococcus lactis (Shandong Baolai Lailai Biotechnology Co., Ltd., ≥

[0034] 3.0×10 9 CFU / g).

[0035] Example 1

[0036] 1) Crush the rice bran meal through a 20-mesh sieve, and take the powder that passes through the sieve for steam explosion treatment (Changchun Peterford Technology Co., Ltd., BTF-088). The reaction conditions are: steam pressure 0.5MPa, temperature 120℃, pressure holding time 1min, gas release (explosion) time 150ms, and rice bran meal moisture content 15%.

[0037] 2) Add 1.0 mol / L hydrochloric acid to the steam-exploded rice bran for acidification treatment. The mass ratio of the steam-exploded rice bran to hydrochloric acid is 4:3. React at room temperature for 30 min to obtain modified rice bran.

[0038] 3) The modified rice bran meal was mixed with phytase, xylanase, cellulase, saccharifying enzyme, and acidic protease and stirred evenly. The mixture was reacted at 40℃ for 3 hours. The amount of phytase added was 100 U / g modified rice bran meal, the amount of xylanase added was 10 U / g modified rice bran meal, the amount of cellulase added was 1500 U / g modified rice bran meal, the amount of saccharifying enzyme added was 50 U / g modified rice bran meal, and the amount of acidic protease added was 250 U / g modified rice bran meal. After the reaction was completed, the mixture was dried at 65℃ for 6 hours to obtain functionalized rice bran meal.

[0039] 4) Mix copper sulfate pentahydrate, zinc sulfate monohydrate, glycine, calcium carbonate and water in a mass ratio of copper ions, zinc ions, glycine, calcium carbonate and water of 1:2:6:4:50. Stir the mixture at 30°C until no more bubbles are produced, indicating the end of the reaction (the reaction time is 0.5 h). Centrifuge at 3000 r / min for 5 min and collect the supernatant to obtain a copper ion + zinc ion solution.

[0040] 5) Mix functionalized rice bran meal with copper ion + zinc ion solution evenly, add lactic acid bacteria liquid, the mass ratio of functionalized rice bran meal, copper ion + zinc ion solution and lactic acid bacteria liquid is 20:15:2, ferment at 25℃ for 12h, after the reaction is completed, dry at 50℃ for 4h to obtain high-efficiency copper-zinc chelate composite material.

[0041] The process of activating and obtaining lactic acid bacteria culture:

[0042] Add 10% brown sugar to sterile distilled water by weight, stir until completely dissolved, then add 1% bacterial powder by weight of the sugar solution, stir slowly and evenly, seal, and incubate at 37℃ for 5 hours to obtain a lactic acid bacteria solution. The pH of the activated lactic acid bacteria solution is 3.5. The concentration of live lactic acid bacteria in the solution is 6 × 10⁻⁶. 8 CFU / mL.

[0043] Example 2 (Preferred Example)

[0044] 1) Crush the rice bran meal through a 30-mesh sieve, and take the powder that passes through the sieve for steam explosion treatment (Changchun Peterford Technology Co., Ltd., BTF-088). The reaction conditions are: steam pressure 0.6MPa, temperature 130℃, pressure holding time 2min, gas release (explosion) time 180ms, and rice bran meal moisture content 20%.

[0045] 2) Add 1.1 mol / L hydrochloric acid to the steam-exploded rice bran for acidification treatment. The mass ratio of the steam-exploded rice bran to hydrochloric acid is 4:4. React at room temperature for 40 min to obtain modified rice bran.

[0046] 3) The modified rice bran meal was mixed with phytase, xylanase, cellulase, saccharifying enzyme, and acidic protease and stirred evenly. The mixture was reacted at 45℃ for 4 hours. The amount of phytase added was 150 U / g modified rice bran meal, the amount of xylanase added was 50 U / g modified rice bran meal, the amount of cellulase added was 2200 U / g modified rice bran meal, the amount of saccharifying enzyme added was 100 U / g modified rice bran meal, and the amount of acidic protease added was 500 U / g modified rice bran meal. After the reaction was completed, the mixture was dried at 75℃ for 8 hours to obtain functionalized rice bran meal.

[0047] 4) Mix copper sulfate pentahydrate, zinc sulfate monohydrate, glycine, calcium carbonate and water in a mass ratio of copper ions, zinc ions, glycine, calcium carbonate and water of 1:4:8:5:80. Stir the mixture at 40°C until no more bubbles are produced in the system, indicating that the reaction is complete (the reaction time is 1 hour). Centrifuge at 4000 rpm for 6 minutes and collect the supernatant to obtain a copper ion + zinc ion solution.

[0048] 5) Mix functionalized rice bran meal with copper ion + zinc ion solution evenly, add lactic acid bacteria liquid, the mass ratio of functionalized rice bran meal, copper ion + zinc ion solution and lactic acid bacteria liquid is 20:20:3, ferment at 30℃ for 24h, after the reaction is completed, dry at 55℃ for 6h to obtain high-efficiency copper-zinc chelate composite material.

[0049] The process of activating and obtaining lactic acid bacteria culture:

[0050] Add 10% brown sugar to sterile distilled water by weight and stir until completely dissolved. Add 1% bacterial powder by weight of the sugar solution, stir slowly and evenly, seal, and incubate at 37℃ for 4.5 hours to obtain a lactic acid bacteria solution. The pH of the activated lactic acid bacteria solution is 4. The concentration of live lactic acid bacteria in the solution is 1×10⁻⁶. 8 CFU / mL.

[0051] Example 3 (More preferred example)

[0052] 1) The rice bran meal was crushed and passed through a 40-mesh sieve. The powder that passed through the sieve was subjected to steam explosion treatment (Changchun Peterford Technology Co., Ltd., BTF-088). The reaction conditions were: steam pressure 0.8MPa, temperature 150℃, pressure holding time 3min, venting (explosion) time 250ms, and the moisture content of the rice bran meal was 22.5%.

[0053] 2) Add 1.25 mol / L hydrochloric acid to the steam-exploded rice bran for acidification treatment. The mass ratio of the steam-exploded rice bran to hydrochloric acid is 4:5.5. React at room temperature for 45 min to obtain modified rice bran.

[0054] 3) The modified rice bran meal was mixed with phytase, xylanase, cellulase, saccharifying enzyme, and acidic protease and stirred evenly. The mixture was reacted at 50℃ for 5 hours. The amount of phytase added was 170 U / g modified rice bran meal, the amount of xylanase added was 200 U / g modified rice bran meal, the amount of cellulase added was 2500 U / g modified rice bran meal, the amount of saccharifying enzyme added was 130 U / g modified rice bran meal, and the amount of acidic protease added was 700 U / g modified rice bran meal. After the reaction was completed, the mixture was dried at 80℃ for 12 hours to obtain functionalized rice bran meal.

[0055] 4) Mix copper sulfate pentahydrate, zinc sulfate monohydrate, glycine, calcium carbonate and water in a mass ratio of copper ions, zinc ions, glycine, calcium carbonate and water of 1:8:11:7:110. Stir the mixture at 45°C until no more bubbles are produced, indicating the end of the reaction (the reaction time is 1.5 h). Centrifuge at 5000 r / min for 9 min and collect the supernatant to obtain a copper ion + zinc ion solution.

[0056] 5) Mix functionalized rice bran meal with copper ion + zinc ion solution evenly, add lactic acid bacteria liquid, the mass ratio of functionalized rice bran meal, copper ion + zinc ion solution and lactic acid bacteria liquid is 20:22:4, ferment at 34℃ for 30h, after the reaction is completed, dry at 60℃ for 10h to obtain high-efficiency copper-zinc chelate composite material.

[0057] The process of activating and obtaining lactic acid bacteria culture:

[0058] Add 10% brown sugar to sterile distilled water by weight, stir until completely dissolved, then add 1% bacterial powder by weight of the sugar solution, stir slowly and evenly, seal, and incubate at 37℃ for 4 hours to obtain a lactic acid bacteria solution. The pH of the activated lactic acid bacteria solution is 4.2. The concentration of live lactic acid bacteria in the solution is 4 × 10⁻⁶. 7 CFU / mL.

[0059] Example 4 (Preferred Example)

[0060] 1) The rice bran meal was crushed and passed through a 50-mesh sieve. The powder that passed through the sieve was subjected to steam explosion treatment (Changchun Peterford Technology Co., Ltd., BTF-088). The reaction conditions were: steam pressure 1.0 MPa, temperature 170℃, pressure holding time 4 min, gas release (explosion) time 320 ms, and rice bran meal moisture content 25%.

[0061] 2) Add 1.4 mol / L hydrochloric acid to the steam-exploded rice bran meal for acidification treatment. The mass ratio of the steam-exploded rice bran meal to hydrochloric acid is 4:7. React at room temperature for 50 min to obtain modified rice bran meal.

[0062] 3) The modified rice bran meal was mixed with phytase, xylanase, cellulase, saccharifying enzyme, and acidic protease and stirred evenly. The mixture was reacted at 55℃ for 6 hours. The amount of phytase added was 200 U / g modified rice bran meal, the amount of xylanase added was 500 U / g modified rice bran meal, the amount of cellulase added was 2800 U / g modified rice bran meal, the amount of saccharifying enzyme added was 150 U / g modified rice bran meal, and the amount of acidic protease added was 1000 U / g modified rice bran meal. After the reaction was completed, the mixture was dried at 85℃ for 20 hours to obtain functionalized rice bran meal.

[0063] 4) Mix copper sulfate pentahydrate, zinc sulfate monohydrate, glycine, calcium carbonate and water in a mass ratio of copper ions, zinc ions, glycine, calcium carbonate and water of 1:12:13:9:150. Stir the mixture at 50°C until no more bubbles are produced in the system, indicating that the reaction is complete (the reaction time is 2 hours). Centrifuge at 7000 r / min for 12 minutes and collect the supernatant to obtain a copper ion + zinc ion solution.

[0064] 5) Mix functionalized rice bran meal with copper ion + zinc ion solution evenly, add lactic acid bacteria liquid, the mass ratio of functionalized rice bran meal, copper ion + zinc ion solution and lactic acid bacteria liquid is 20:25:5, ferment at 37℃ for 48h, after the reaction is completed, dry at 65℃ for 20h to obtain high-efficiency copper-zinc chelate composite material.

[0065] The process of activating and obtaining lactic acid bacteria culture:

[0066] Add 10% brown sugar to sterile distilled water by weight and stir until completely dissolved. Add 1% bacterial powder by weight of the sugar solution, stir slowly and evenly, seal, and incubate at 37℃ for 3.5 hours to obtain a lactic acid bacteria solution. The pH of the activated lactic acid bacteria solution is 4.5. The concentration of live lactic acid bacteria in the solution is 1×10⁻⁶. 7 CFU / mL.

[0067] Example 5

[0068] 1) The rice bran meal was crushed and passed through a 60-mesh sieve. The powder that passed through the sieve was subjected to steam explosion treatment (Changchun Peterford Technology Co., Ltd., BTF-088). The reaction conditions were: steam pressure 1.2MPa, temperature 180℃, pressure holding time 5min, venting (explosion) time 350ms, and rice bran meal moisture content 30%.

[0069] 2) Add 1.5 mol / L hydrochloric acid to the steam-exploded rice bran for acidification treatment. The mass ratio of the steam-exploded rice bran to hydrochloric acid is 4:8. React at room temperature for 60 min to obtain modified rice bran.

[0070] 3) The modified rice bran meal was mixed with phytase, xylanase, cellulase, saccharifying enzyme, and acidic protease and stirred evenly. The mixture was reacted at 60℃ for 7 hours. The amount of phytase added was 250 U / g modified rice bran meal, the amount of xylanase added was 800 U / g modified rice bran meal, the amount of cellulase added was 3500 U / g modified rice bran meal, the amount of saccharifying enzyme added was 200 U / g modified rice bran meal, and the amount of acidic protease added was 2000 U / g modified rice bran meal. After the reaction was completed, the mixture was dried at 95℃ for 24 hours to obtain functionalized rice bran meal.

[0071] 4) Mix copper sulfate pentahydrate, zinc sulfate monohydrate, glycine, calcium carbonate and water in a mass ratio of copper ions, zinc ions, glycine, calcium carbonate and water of 1:15:15:10:200. Stir the mixture at 60°C until no more bubbles are produced, indicating the end of the reaction (the reaction time is 2.5 h). Centrifuge at 8000 r / min for 15 min and collect the supernatant to obtain a copper ion + zinc ion solution.

[0072] 5) Mix functionalized rice bran meal with copper ion + zinc ion solution evenly, add lactic acid bacteria liquid, the mass ratio of functionalized rice bran meal, copper ion + zinc ion solution and lactic acid bacteria liquid is 20:30:6, ferment at 40℃ for 72h, after the reaction is completed, dry at 65℃ for 24h to obtain high-efficiency copper-zinc chelate composite material.

[0073] The process of activating and obtaining lactic acid bacteria culture:

[0074] Add 10% brown sugar to sterile distilled water by weight, stir until completely dissolved, then add 1% bacterial powder by weight of the sugar solution, stir slowly and evenly, seal, and incubate at 37℃ for 3 hours to obtain a lactic acid bacteria solution; the pH of the activated lactic acid bacteria solution is 5. The concentration of live lactic acid bacteria in the solution is 5 × 10⁻⁶. 6 CFU / mL.

[0075] Indicator Test

[0076] 1. Determination of polysaccharide and protein indicators in rice bran meal before and after modification

[0077] Method: The modified rice bran meal prepared in steps 1)-2) of each embodiment was dried at 85℃ for 12 hours, and then simultaneously subjected to water dissolution treatment with the rice bran meal raw material. The operation steps are as follows:

[0078] Take 10g of rice bran meal before modification and 10g of rice bran meal after modification (steam explosion (i.e., step 1 in the example, the same below) + acidification treatment (i.e., step 2 in the example, the same below)). Add 200mL of water and dissolve in water at room temperature for 1h, boil at 90℃ for 1h, and boil at 90℃ for 2h, respectively. Centrifuge, and add amylase (Henan Zhongchen Biotechnology Co., Ltd., high-temperature resistant α-amylase, 1000g pack) to the supernatant at 1% of the raw material mass ratio for starch and dextrin removal treatment at 65℃. Every 0.5 hours, take 1 mL of supernatant and add 2 drops of KI-I2 reagent. Observe the color change. After the color changes from blue to purple or red, take 5 mL of supernatant, add 20 mL of anhydrous ethanol, refrigerate overnight (12 hours), centrifuge, and dilute the precipitate with 50 mL of water. Take 1 mL to determine the polysaccharide content according to the standard "SN / T4260-2015 Determination of Crude Polysaccharides in Exported Plant-Derived Foods - Phenol-Sulfuric Acid Method". Calculate the polysaccharide content in the supernatant. At the same time, take 2 mL of supernatant to determine the protein content according to "GB 5009.5-2025 National Food Safety Standard - Determination of Protein in Food".

[0079] After drying the precipitates corresponding to each treatment, residue samples were obtained. 5g of each residue sample was taken, 50mL of water was added, and the mixture was boiled at 90℃ for 2 hours. The solution was then diluted to 100mL in a volumetric flask, centrifuged, and the supernatant was used to determine the polysaccharide and protein content again using the method described above. Data comparison is shown in the appendix. Figure 1 The determination of polysaccharide and protein in rice bran meal before and after modification at room temperature for 1 hour, and related data are attached. Figure 2 Determination of polysaccharide and protein in rice bran meal after 1 hour of boiling under heating conditions before and after modification in each embodiment. Figure 3 The polysaccharide and protein indices of rice bran meal were determined by boiling in water for 2 hours before and after modification in each embodiment.

[0080] Depend on Figure 1The data shows that the structure of rice bran meal changed significantly after steam explosion and acidification modification. This treatment allows the key active substances that play a chelating role in rice bran meal, namely polysaccharides and proteins, to be transferred from the residue to the supernatant. After water dissolution at room temperature for 1 hour, the polysaccharide content in the supernatant increased from 0.63% to 3.05%, and the protein content increased from 2.87% to 4.41%.

[0081] Depend on Figure 2 The data shows that heat treatment can promote the dissolution of polysaccharides and proteins. After boiling in water at 90℃ for 1 hour, the polysaccharide content in the supernatant increased from 1.58% to 4.68%, and the protein content increased from 3.78% to 4.95%.

[0082] Depend on Figure 3 The data shows that extending the heating time can further promote the dissolution of polysaccharides and proteins. After boiling in water at 90℃ for 2 hours, the polysaccharide content in the supernatant increased from 1.75% to 5.59%, and the protein content increased from 4.82% to 6.09%.

[0083] 2. Determination of polysaccharide and protein peptide indices before and after functionalization treatment of rice bran meal

[0084] The sample processing method is as follows:

[0085] The rice bran meal raw material (rice bran meal is crushed and passed through a 40-mesh sieve, and the powder that passes through the sieve is the rice bran meal raw material) is marked as the control sample;

[0086] Rice bran meal without steam explosion and acidification modification was processed according to the functionalization steps in each embodiment (the process and conditions are the same as step 3 in each embodiment), except that rice bran meal raw material was used instead of modified rice bran meal, and samples 1 of each embodiment were obtained respectively.

[0087] The rice bran meal raw material was processed according to the steam explosion + acidification modification steps in each embodiment (i.e., steps 1)-2) to obtain the modified rice bran meal, and samples 2 of each embodiment were obtained respectively.

[0088] The rice bran meal raw material was processed according to the steam explosion + acidification modification steps and functionalization treatment steps in each embodiment (i.e., steps 1)-3 in each embodiment to obtain functionalized rice bran meal), and samples 3 of each embodiment were obtained respectively.

[0089] Methods for determining polysaccharide indexes:

[0090] The above samples were extracted at a material-to-water mass ratio of 1:20 at room temperature for 10 min. After centrifugation, the supernatant was obtained. Four volumes of anhydrous ethanol were added to the supernatant, and the mixture was refrigerated overnight (12 hours). After centrifugation, the precipitate was dried at 60℃ for 8 h to obtain crude rice bran polysaccharide. The yield (polysaccharide yield was calculated according to the formula below) and content (polysaccharide content was determined according to the standard "SN / T 4260-2015 Determination of Crude Polysaccharides in Exported Plant-Derived Foods by Phenol-Sulfuric Acid Method") of rice bran polysaccharide extracted from different samples at room temperature for a short time were determined. The polysaccharide extraction rate was calculated (polysaccharide extraction rate = yield × content / 100).

[0091] Polysaccharide yield = m1 / m2 × 100%

[0092] In the formula: m1 is the mass of crude rice bran polysaccharide, g; m2 is the mass of the sample, g.

[0093] To eliminate the interference of starch and dextrin on the determination of polysaccharide content, samples containing starch and dextrin were treated to remove starch and dextrin before the determination of polysaccharide content (the specific operation is the same as the starch and dextrin removal process in section 1 above for the determination of polysaccharide and protein indicators before and after rice bran meal modification). Data comparison is shown in [link to data]. Figure 4 Determination of polysaccharide indexes of rice bran meal before and after functionalization treatment in each embodiment.

[0094] After functionalization, the nutritional composition of modified rice bran meal changes further, providing a material basis for a significant improvement in chelation capacity.

[0095] Depend on Figure 4 The data shows that rice bran meal that has undergone steam explosion + acidification modification and functionalization treatment can achieve the following results after only 10 minutes of extraction at room temperature: Example 1: Polysaccharide extraction rate increased from 0% to 7.14%; Example 2: Polysaccharide extraction rate increased to 10.9%; Example 3: Polysaccharide extraction rate increased to 14.5%; Example 4: Polysaccharide extraction rate increased to 12.9%; Example 5: Polysaccharide extraction rate increased to 8.46%. This demonstrates that both modification and functionalization treatments are indispensable.

[0096] Methods for determining protein peptide indicators:

[0097] The acid-soluble protein content and peptide molecular weight composition of the above samples were determined according to GB / T 22492-2008 Soybean Peptide Powder. Data comparison is shown below. Figure 5 Determination of protein peptide indexes before and after functionalization treatment of rice bran meal in each embodiment.

[0098] Depend on Figure 5The data shows that, in rice bran meal that has undergone steam explosion + acidification modification and functionalization treatment simultaneously, the acid-soluble protein content in Example 1 increased from 1.33% in the control to 12.4%, and the proportion of small peptides below 1000 Da increased from 8.9% in the control to 80.7%; in Example 2, the acid-soluble protein content increased to 13.1%, and the proportion of small peptides below 1000 Da increased to 86.6%; in Example 3, the acid-soluble protein content increased to 14.2%, and the proportion of small peptides below 1000 Da increased to 90.3%; in Example 4, the acid-soluble protein content increased to 13.6%, and the proportion of small peptides below 1000 Da increased to 88.2%; and in Example 5, the acid-soluble protein content increased to 12.7%, and the proportion of small peptides below 1000 Da increased to 83.8%. This indicates that both modification and functionalization treatments are indispensable.

[0099] Comparative Example 1

[0100] Treatment Group 1 (chelation of rice bran meal raw material, without modification, functionalization, metal ion activation, or fermentation treatment)

[0101] Copper sulfate pentahydrate and zinc sulfate monohydrate were mixed with water at a mass ratio of copper ions, zinc ions, and water of 1:2:50. The mixture was stirred at 30°C for 0.5 hours, centrifuged at 3000 rpm for 5 minutes, and the supernatant was collected to obtain a copper ion + zinc ion solution. Rice bran was pulverized and passed through a 20-mesh sieve. The powder that passed through the sieve was mixed evenly with the copper ion + zinc ion solution at a mass ratio of 20:15. The mixture was reacted at 25°C for 12 hours. After the reaction, the mixture was dried at 50°C for 4 hours to obtain sample No. 1 of treatment group 1 (Comparative Example 1).

[0102] Treatment Group 2 (functionalized treatment without rice bran meal)

[0103] The process and conditions are the same as steps 1), 2), 4), and 5) in Example 1, except that step 3) is not performed, and the functionalized rice bran meal in step 5) is replaced with the modified rice bran meal in step 2) to obtain sample No. 2 of the treatment group of Comparative Example 1.

[0104] Treatment Group 3 (Modification treatment without rice bran meal)

[0105] The rice bran meal was crushed and passed through a 20-mesh sieve. The powder that passed through the sieve was then processed according to step 3 (using rice bran meal as raw material instead of modified rice bran meal), step 4), and step 5 in Example 1 to obtain sample No. 3 of the treatment group of Comparative Example 1.

[0106] Treatment group 4 (activation treatment without metal ions)

[0107] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 1, except that glycine and calcium carbonate are not added in step 4), resulting in sample No. 4 of the treatment group of Comparative Example 1.

[0108] Treatment group 5 (no fermentation treatment)

[0109] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 1, except that no lactic acid bacteria solution is added in step 5), resulting in sample No. 5 of the treatment group of Comparative Example 1.

[0110] Comparative Example 2

[0111] Treatment Group 1 (chelation of rice bran meal raw material, without modification, functionalization, metal ion activation, or fermentation treatment)

[0112] Copper sulfate pentahydrate and zinc sulfate monohydrate were mixed with water at a mass ratio of copper ions, zinc ions, and water of 1:4:80. The mixture was stirred at 40°C for 1 hour, centrifuged at 4000 rpm for 6 minutes, and the supernatant was collected to obtain a copper ion + zinc ion solution. Rice bran was pulverized and passed through a 30-mesh sieve. The powder that passed through the sieve was mixed evenly with the copper ion + zinc ion solution at a mass ratio of 20:20. The mixture was reacted at 30°C for 24 hours. After the reaction was completed, the mixture was dried at 55°C for 6 hours to obtain sample No. 1 of treatment group 2.

[0113] Treatment Group 2 (functionalized treatment without rice bran meal)

[0114] The process and conditions are the same as steps 1), 2), 4), and 5) in Example 2, except that step 3) is not performed, and the functionalized rice bran meal in step 5) is replaced with the modified rice bran meal in step 2) to obtain sample No. 2 of the treatment group of Comparative Example 2.

[0115] Treatment Group 3 (Modification treatment without rice bran meal)

[0116] The rice bran meal was crushed and passed through a 30-mesh sieve. The powder that passed through the sieve was then processed according to step 3 (using rice bran meal as raw material instead of modified rice bran meal), step 4), and step 5 in Example 2 to obtain sample No. 3 of the treatment group of Comparative Example 2.

[0117] Treatment group 4 (activation treatment without metal ions)

[0118] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 2, except that glycine and calcium carbonate are not added in step 4), resulting in sample No. 4 of the treatment group of Comparative Example 2.

[0119] Treatment group 5 (no fermentation treatment)

[0120] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 2, except that no lactic acid bacteria solution is added in step 5), resulting in sample No. 5 of the treatment group of Comparative Example 2.

[0121] Comparative Example 3

[0122] Treatment Group 1 (chelation of rice bran meal raw material, without modification, functionalization, metal ion activation, or fermentation treatment)

[0123] Copper sulfate pentahydrate and zinc sulfate monohydrate were mixed with water at a mass ratio of copper ions, zinc ions, and water of 1:8:110. The mixture was stirred at 45°C for 1.5 hours, centrifuged at 5000 rpm for 9 minutes, and the supernatant was collected to obtain a copper ion + zinc ion solution. Rice bran meal was pulverized and passed through a 40-mesh sieve. The powder that passed through the sieve was mixed evenly with the copper ion + zinc ion solution at a mass ratio of 20:22. The mixture was reacted at 34°C for 30 hours. After the reaction, the mixture was dried at 60°C for 10 hours to obtain sample No. 1 of treatment group 3.

[0124] Treatment Group 2 (functionalized treatment without rice bran meal)

[0125] The process and conditions are the same as steps 1), 2), 4), and 5) in Example 3, except that step 3) was not performed, and the functionalized rice bran meal in step 5) was replaced with the modified rice bran meal in step 2) to obtain sample No. 2 of the treatment group of Comparative Example 3.

[0126] Treatment Group 3 (Modification treatment without rice bran meal)

[0127] The rice bran meal was crushed and passed through a 40-mesh sieve. The powder that passed through the sieve was then processed according to step 3 (using rice bran meal as raw material instead of modified rice bran meal), step 4), and step 5 in Example 3 to obtain sample No. 3 of the treatment group of Comparative Example 3.

[0128] Treatment group 4 (activation treatment without metal ions)

[0129] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 3, except that glycine and calcium carbonate are not added in step 4), resulting in sample No. 4 of the treatment group of Comparative Example 3.

[0130] Treatment group 5 (no fermentation treatment)

[0131] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 3, except that no lactic acid bacteria solution is added in step 5), resulting in sample No. 5 of the treatment group of Comparative Example 3.

[0132] Comparative Example 4

[0133] Treatment Group 1 (chelation of rice bran meal raw material, without modification, functionalization, metal ion activation, or fermentation treatment)

[0134] Copper sulfate pentahydrate and zinc sulfate monohydrate were mixed with water at a mass ratio of copper ions, zinc ions, and water of 1:12:150. The mixture was stirred at 50°C for 2 hours, centrifuged at 7000 rpm for 12 minutes, and the supernatant was collected to obtain a copper ion + zinc ion solution. Rice bran was pulverized and passed through a 50-mesh sieve. The powder that passed through the sieve was mixed evenly with the copper ion + zinc ion solution at a mass ratio of 20:25. The mixture was reacted at 37°C for 48 hours. After the reaction, the mixture was dried at 65°C for 20 hours to obtain sample No. 1 of treatment group 4.

[0135] Treatment Group 2 (functionalized treatment without rice bran meal)

[0136] The process and conditions are the same as steps 1), 2), 4), and 5) in Example 4, except that step 3) was not performed, and the functionalized rice bran meal in step 5) was replaced with the modified rice bran meal in step 2) to obtain sample No. 2 of the treatment group in Comparative Example 4.

[0137] Treatment Group 3 (Modification treatment without rice bran meal)

[0138] The rice bran meal was crushed and passed through a 50-mesh sieve. The powder that passed through the sieve was then processed according to step 3 (using rice bran meal as raw material instead of modified rice bran meal), step 4, and step 5 in Example 4 to obtain sample No. 3 of the treatment group of Comparative Example 4.

[0139] Treatment group 4 (activation treatment without metal ions)

[0140] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 4, except that glycine and calcium carbonate are not added in step 4), resulting in sample No. 4 of the treatment group of Comparative Example 4.

[0141] Treatment group 5 (no fermentation treatment)

[0142] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 4, except that no lactic acid bacteria solution is added in step 5), resulting in sample No. 5 of the treatment group of Comparative Example 4.

[0143] Comparative Example 5

[0144] Treatment Group 1 (chelation of rice bran meal raw material, without modification, functionalization, metal ion activation, or fermentation treatment)

[0145] Copper sulfate pentahydrate and zinc sulfate monohydrate were mixed with water at a mass ratio of copper ions, zinc ions, and water of 1:15:200. The mixture was stirred at 60°C for 2.5 hours, centrifuged at 8000 rpm for 15 minutes, and the supernatant was collected to obtain a copper ion + zinc ion solution. Rice bran meal was pulverized and passed through a 60-mesh sieve. The powder that passed through the sieve was mixed evenly with the copper ion + zinc ion solution at a mass ratio of 20:30. The mixture was reacted at 40°C for 72 hours. After the reaction, the mixture was dried at 65°C for 24 hours to obtain sample No. 1 of treatment group 5 (Comparative Example 5).

[0146] Treatment Group 2 (functionalized treatment without rice bran meal)

[0147] The process and conditions are the same as steps 1), 2), 4), and 5) in Example 5, except that step 3) is not performed, and the functionalized rice bran meal in step 5) is replaced with the modified rice bran meal in step 2) to obtain sample No. 2 of treatment group 5.

[0148] Treatment Group 3 (Modification treatment without rice bran meal)

[0149] The rice bran meal was crushed and passed through a 60-mesh sieve. The powder that passed through the sieve was then processed according to step 3 (using rice bran meal as raw material instead of modified rice bran meal), step 4), and step 5 in Example 5 to obtain sample No. 3 of the treatment group of Comparative Example 5.

[0150] Treatment group 4 (activation treatment without metal ions)

[0151] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 5, except that glycine and calcium carbonate are not added in step 4), resulting in sample No. 4 of the treatment group of Comparative Example 5.

[0152] Treatment group 5 (no fermentation treatment)

[0153] The process and conditions are the same as steps 1), 2), 3), 4), and 5) in Example 5, except that no lactic acid bacteria solution is added in step 5), resulting in sample No. 5 of the treatment group of Comparative Example 5.

[0154] Test case

[0155] (1) Chelation rate test

[0156] Test Method Description

[0157] The commonly used method in the industry for determining chelation rate is the organic solvent dissolution-EDTA titration method. This method is based on the principle that trace element chelates are insoluble in organic solvents, while free metal ions are soluble in organic solvents, and uses this property to calculate the chelation rate of the sample. When a metal ion itself is insoluble in an organic solvent, it will be included in V. 螯合 This can interfere with the measurement results. During the experiment, the inventors discovered that among many influencing factors, the drying temperature of the chelated sample was the most significant factor interfering with the chelation rate measurement. Therefore, in order to accurately reflect the chelation rate of the sample, this invention limited the drying temperature of the high-efficiency copper-zinc chelated composite material to 50-65℃. Above this temperature, the measurement results cannot indicate the true chelation rate. The copper-zinc chelation rate measurement results of the comparative examples and embodiments are shown in the table below:

[0158] Table 1 Comparison of copper chelation rates of composite materials prepared in comparative examples and embodiments

[0159]

[0160] The data in the table shows that in the preparation process of high-efficiency copper-zinc chelate composite materials, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are all missing, the copper chelation rate is <30%; if any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the copper chelation rate cannot reach more than 90%, indicating that the steps described in the examples are indispensable in the preparation process of high-efficiency copper-zinc chelate composite materials.

[0161] Table 2 Comparison of zinc chelation rates of composite materials prepared in comparative examples and embodiments

[0162]

[0163] The data in the table shows that in the preparation of high-efficiency copper-zinc chelate composite materials, if the steps of rice bran modification, rice bran functionalization, metal ion activation, and fermentation are missing, the zinc chelation rate can only be maintained at about 20%. If any one of the steps of rice bran modification, rice bran functionalization, metal ion activation, or fermentation is missing, the zinc chelation rate cannot reach more than 90%. This indicates that the steps described in the examples are indispensable in the preparation of high-efficiency copper-zinc chelate composite materials.

[0164] (2) Copper-zinc ion release rate test

[0165] The in vitro simulated gastrointestinal digestion experiment followed the method described in Li Congmiao's Master's Thesis, "Preparation of Oligopeptide-Copper / Zinc Chelates by Solid-State Fermentation of Soybean Meal and Characterization of Their Structural Properties," published in 2023 at Nanchang University. 120 mg of pepsin was dissolved in 100 mL of ultrapure water to prepare simulated gastric juice (SGF), and 144 mg of trypsin and 864 mg of bile salts were dissolved in 0.1 mol / L NaHCO3 solution to prepare simulated intestinal juice (SIF). A solution of the high-efficiency copper-zinc chelate composite material was prepared at a concentration of 2.5 mg / mL. The pH of the solution was adjusted to 2.0 with 1 mol / L HCl. SGF (enzyme to substrate mass ratio 1:50, w / w) was added, and the solution was incubated at 37°C with shaking for 120 min. The pH was then adjusted to 7.5, and SIF (enzyme to substrate mass ratio 1:25, w / w) was added to initiate the intestinal digestion stage. The solution was incubated at 37°C with shaking for 120 min. Gastric and intestinal digestive fluids collected at 60 and 120 min were heated at 100℃ for 10 min to inactivate enzymes. The fluids were then aliquoted and centrifuged at 8000 r / min for 15 min. The supernatant was used to determine the copper and zinc content.

[0166] Method for determining metal release rate

[0167] Take 5 mL of supernatant and mix it with 10 mL of nitric acid in a digestion tube, and digest it using a graphite digester. After removing the acid and making up the volume, the copper and zinc contents of the digested solution are determined by an ICP-MS analyzer. The solution is filtered through a 0.22 μm filter membrane before being processed.

[0168] Metal release rate (%) = m1 / m × 100

[0169] In the formula: m represents the total copper and total zinc content (mg) in the high-efficiency copper-zinc chelate composite material, and m1 is the free copper and zinc content (mg) in the supernatant.

[0170] Table 3 Comparison of copper release rates of the composite materials prepared in the comparative examples and the actual examples after 60 min in simulated gastric fluid.

[0171]

[0172] Highly effective mineral supplements require protection for metal ions as they pass through the stomach. If chelates dissociate significantly in the stomach, the released metal ions will, on the one hand, combine with other substances such as phytic acid and oxalic acid to form insoluble precipitates that cannot be absorbed; on the other hand, different metal ions compete with each other for absorption, and premature dissociation increases this competition, reducing the absorption rate of the target mineral. Furthermore, free metal ions can irritate the gastric mucosa, producing a series of side effects.

[0173] The data in the table shows that in the preparation process of the high-efficiency copper-zinc chelate composite material, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are all missing, the copper release rate after 60 minutes of simulated gastric juice is as high as 70% or more. If any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the copper release rate after 60 minutes of simulated gastric juice cannot be reduced to below 10%. This indicates that the steps described in the examples are indispensable in the preparation process of the high-efficiency copper-zinc chelate composite material.

[0174] Table 4 Comparison of copper release rates of the composite materials prepared in the comparative examples and the actual examples after 120 min from simulated gastric fluid.

[0175]

[0176]

[0177] The data in the table shows that in the preparation process of the high-efficiency copper-zinc chelate composite material, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are missing, the copper release rate after 120 minutes of simulated gastric juice is as high as 80% or more. If any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the copper release rate after 120 minutes of simulated gastric juice cannot be controlled at around 10%. This indicates that the steps described in the examples are indispensable in the preparation process of the high-efficiency copper-zinc chelate composite material.

[0178] Table 5 Comparison of copper release rates of the composite materials prepared in the comparative examples and the actual examples after 60 min of simulated intestinal fluid.

[0179]

[0180] The small intestine (especially the duodenum and jejunum) is the primary site of mineral absorption. Effective mineral supplements require targeted release and absorption within the intestines.

[0181] The data in the table shows that in the preparation process of high-efficiency copper-zinc chelate composite materials, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are missing, the copper release rate after 60 minutes of simulated intestinal fluid is only about 20%. If any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the copper release rate after 60 minutes of simulated intestinal fluid cannot reach more than 80%. This indicates that the steps described in the examples are indispensable in the preparation process of high-efficiency copper-zinc chelate composite materials.

[0182] Table 6 Comparison of copper release rates of the composite materials prepared in the comparative examples and the actual examples after 120 min of simulated intestinal fluid.

[0183]

[0184]

[0185] The data in the table shows that in the preparation process of high-efficiency copper-zinc chelate composite materials, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are missing, the copper release rate after 120 minutes of simulated intestinal fluid will still remain at around 20%. If any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the copper release rate after 120 minutes of simulated intestinal fluid cannot reach more than 80%, indicating that the steps described in the examples are indispensable in the preparation process of high-efficiency copper-zinc chelate composite materials.

[0186] Table 7 Comparison of zinc release rates of the composite materials prepared in the comparative examples and the actual examples after 60 min of simulating gastric fluid.

[0187]

[0188] The data in the table shows that in the preparation process of the high-efficiency copper-zinc chelate composite material, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are missing, the zinc release rate after 60 minutes of simulated gastric juice is as high as 65% or more. If any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the zinc release rate after 60 minutes of simulated gastric juice cannot be reduced to below 10%. This indicates that the steps described in the examples are indispensable in the preparation process of the high-efficiency copper-zinc chelate composite material.

[0189] Table 8 Comparison of zinc release rates of the composite materials prepared in the comparative examples and embodiments after 120 min from simulated gastric fluid.

[0190]

[0191]

[0192] The data in the table shows that in the preparation process of the high-efficiency copper-zinc chelate composite material, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are missing, the zinc release rate after 120 minutes of simulated gastric juice is as high as 75% or more. If any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the zinc release rate after 120 minutes of simulated gastric juice cannot be reduced to below 10%. This indicates that the steps described in the examples are indispensable in the preparation process of the high-efficiency copper-zinc chelate composite material.

[0193] Table 9 Comparison of zinc release rates of simulated intestinal fluid prepared from comparative examples and embodiments after 60 min.

[0194]

[0195] The data in the table shows that in the preparation process of high-efficiency copper-zinc chelate composite materials, if the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment steps are missing, the zinc release rate after 60 minutes of simulated intestinal fluid is only about 30%. If any one of the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, or fermentation treatment steps is missing, the zinc release rate after 60 minutes of simulated intestinal fluid cannot reach more than 75%. This indicates that the steps described in the examples are indispensable in the preparation process of high-efficiency copper-zinc chelate composite materials.

[0196] Table 10 Comparison of zinc release rates of simulated intestinal fluid prepared from comparative examples and embodiments after 120 min.

[0197]

[0198]

[0199] The data in the table show that, in the preparation process of the high-efficiency copper-zinc chelate composite material, even without the rice bran meal modification treatment, rice bran meal functionalization treatment, metal ion activation treatment, and fermentation treatment, the zinc release rate after 120 minutes of simulated intestinal fluid remains at around 35%. Furthermore, without any one of these steps, the zinc release rate after 120 minutes of simulated intestinal fluid cannot reach over 80%, indicating that the steps described in the examples are indispensable in the preparation process of the high-efficiency copper-zinc chelate composite material.

[0200] In summary, all the technical steps in this invention are indispensable.

[0201] This invention utilizes physical, chemical, and biological techniques to modify rice bran, a biomass resource, and combines metal ion activation and microbial fermentation technologies to produce a composite material capable of efficiently chelating and targeting the release of copper and zinc metal ions. This composite material integrates the chelating abilities of various active substances, including polysaccharides, small peptides, oligosaccharides, disaccharides, and organic acids. Through a visualized ion activation process, it achieves multiple chelations of various metal ions, enabling targeted release and multi-channel absorption, thus improving the absorption and utilization rate of copper and zinc trace elements, achieving a highly efficient and safe effect.

Claims

1. A method for preparing a copper-zinc chelate composite material, characterized in that, Includes the following steps: Step (1): Crush the rice bran meal through a 20-60 mesh sieve, and take the powder that passes through the sieve for steam explosion treatment; The reaction conditions are as follows: water vapor pressure 0.5-1.2MPa, temperature 120-180℃, pressure holding time 1-5min, and venting (explosion) time 150-350ms. Step (2): Add 1.0-1.5 mol / L hydrochloric acid to the steam-exploded rice bran meal for acidification treatment to obtain modified rice bran meal; The reaction conditions are as follows: react at room temperature for 30-60 minutes; Step (3): Mix the modified rice bran meal with phytase, xylanase, cellulase, glucoamylase and acidic protease until uniform, and react. After the reaction is completed, dry at 65-95℃ for 6-24 hours to obtain functionalized rice bran meal. The reaction conditions are as follows: react at 40-60℃ for 3-7 hours; Step (4): Mix copper sulfate, zinc sulfate, glycine, calcium carbonate and water, stir and react. After the reaction is complete, centrifuge at 3000-8000 r / min for 5-15 min, collect the supernatant to obtain a copper ion + zinc ion solution. The reaction conditions are: 30-60℃, and the reaction is complete when no more bubbles are produced in the system. Step (5): Mix the functionalized rice bran meal with copper ion + zinc ion solution evenly, add lactic acid bacteria liquid, ferment for 12-72h, after the reaction is completed, dry at 50-65℃ for 4-24h to obtain a high-efficiency copper-zinc chelate composite material. The pH value of the lactic acid bacteria culture is 3.5-5.0, and the fermentation temperature is 25-40℃.

2. The preparation method according to claim 1, characterized in that, In step (1), the moisture content of the rice bran meal during steam explosion treatment is 15-30%.

3. The preparation method according to claim 1, characterized in that, In step (2): during the acidification treatment, the mass ratio of rice bran meal after steam explosion treatment to hydrochloric acid is 4:3-8.

4. The preparation method according to claim 1, characterized in that, In step (3): the amount of phytase added is 100-250 U / g modified rice bran meal, the amount of xylanase added is 10-800 U / g modified rice bran meal, the amount of cellulase added is 1500-3500 U / g modified rice bran meal, the amount of saccharifying enzyme added is 50-200 U / g modified rice bran meal, and the amount of acidic protease added is 250-2000 U / g modified rice bran meal.

5. The preparation method according to claim 1, characterized in that, In step (4), the mass ratio of copper ions, zinc ions, glycine, calcium carbonate, and water is 1:2-15:6-15:4-10:50-200.

6. The preparation method according to claim 1, characterized in that, In step (4): copper sulfate is one or two of anhydrous copper sulfate or copper sulfate pentahydrate, and zinc sulfate is one or two of zinc sulfate monohydrate or zinc sulfate heptahydrate.

7. The preparation method according to claim 1, characterized in that, In step (5): the mass ratio of functionalized rice bran meal, copper-zinc ion solution, and lactic acid bacteria solution is 20:15-30:2-6; the concentration of live lactic acid bacteria in the lactic acid bacteria solution is 10. 6 -10 9 CFU / mL.

8. The preparation method according to claim 1 or 7, characterized in that, In step (5), the fermentation strains of the lactic acid bacteria liquid are commonly used lactic acid bacteria in production, including but not limited to one or more of the genera Lactobacillus, Streptococcus, Enterococcus, and Pediococcus.

9. A copper-zinc chelate composite material prepared by the preparation method according to any one of claims 1-8.

10. The application of the copper-zinc chelate composite material as described in claim 9 in animal feed.

Images