Ampelopsis grossedentata extract compound premix as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of feeds, and particularly relates to an ampelopsis grossedentata extract compound premix as well as a preparation method and application thereof. The compound premix is prepared from the following raw materials in parts by weight: 9 to 15 parts of ampelopsis grossedentata extract, 1 to 5 parts of monostroma extract, 2 to 4 parts of palmitoyl pomegranate peel extract, 1 to 3 parts of modified bentonite, 0.3 to 0.5 part of amino acid and 0.1 to 0.5 part of compound vitamin. The compound premix can significantly improve the animal immunity and the feed conversion rate and reduce the feed conversion ratio.

Description

Technical Field

[0001] This invention belongs to the field of feed technology, specifically relating to a compound premix of Viola yedoensis extract, its preparation method, and its application. Background Technology

[0002] In the current global wave of pursuing green, efficient, and sustainable development in animal husbandry, the feed additive industry is facing unprecedented opportunities and challenges. In the post-antibiotic era, how to ensure animal health and improve production performance through nutritional means has become a core issue for industry development. Among these, the gut, as the largest digestive and absorptive organ and an important immune barrier, directly determines the efficiency of nutrient conversion and the overall immunity of animals. Therefore, developing novel feed additives that can precisely maintain gut health and optimize the gut microbiota is of crucial strategic significance for promoting cost reduction and efficiency improvement in modern animal husbandry.

[0003] In traditional farming practices, farmers often face two key challenges: First, frequent intestinal health problems. Stress, pathogens, and toxins easily damage the integrity of the intestinal mucosa, disrupting the stable microbial community and leading to diseases such as diarrhea and inflammation. This not only increases treatment costs but also severely hinders the growth potential of animals. Second, feed costs remain high, and conversion efficiency needs improvement. Feed accounts for approximately 60%-70% of total farming costs, but a significant portion of nutrients is not effectively digested and absorbed before being excreted, resulting in a high feed conversion ratio that directly erodes farming profits. Previous feed additives have relatively limited functions, focusing either solely on antibacterial activity or supplementing with exogenous probiotics, often failing to systematically address the complex relationship between intestinal structure, microbial balance, and digestion and absorption. To address these industry pain points, improvements to existing compound feed additives are urgently needed. Summary of the Invention

[0004] To overcome the shortcomings of the prior art, the first objective of this invention is to provide a compound premix of Agrostis dentata extract, which can reduce antibiotic use, improve feed conversion rate, reduce feed conversion ratio, increase nutrition, improve meat quality and taste, improve breeding efficiency and reduce costs.

[0005] The second objective of this invention is to provide a simple method for preparing a compound premix of *Agrostis dentata* extract.

[0006] The third objective of this invention is to provide an application of a compound premix of *Agrostis dentata* extract, which has broad prospects.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A compound premix of *Agrostis dentata* extract, comprising the following raw materials in parts by weight: 9-15 parts of *Agrostis dentata* extract, 1-5 parts of reef extract, 2-4 parts of palmitoyl argentea peel extract, 1-3 parts of modified bentonite, 0.3-0.5 parts of amino acids, and 0.1-0.5 parts of compound vitamins.

[0008] Furthermore, the preparation process of the modified bentonite is as follows: (1) Add bentonite and zinc chloride to deionized water, stir at room temperature, and then sonicate. After filtration, washing, and drying, collect product 1. (2) Add potassium hydroxide to deionized water, then add hydroxypropyl guar gum and glycerol in sequence, stir at room temperature and freeze dry, and collect product 2; (3) Add product 1 to deionized water, then add product 2, sonicate and let stand, centrifuge and wash and collect the precipitate; add the precipitate to the crosslinking agent solution for room temperature treatment, and dry to obtain the modified bentonite.

[0009] Furthermore, in step (1), the mass ratio of bentonite, zinc chloride, and deionized water is 1:(0.05-0.08):(6-10); the stirring time is 1-3 hours; the ultrasonic treatment time is 20-30 minutes; and the drying temperature is 110-120°C for 3-5 hours.

[0010] Furthermore, in step (2), the mass ratio of potassium hydroxide, hydroxypropyl guar gum, glycerol, and deionized water is 1:(0.6-1.2):(1.2-2):(20-25); and the stirring time is 40-60 min.

[0011] Further, in step (3), the mass ratio of product 1, product 2, and deionized water is 1:(0.3-0.5):(20-25); the volume ratio of the precipitate to the crosslinking agent solution is 1g:(20-25)mL; the crosslinking agent solution is a calcium chloride solution with a concentration of 10-12wt%; the ultrasonic treatment time is 2.5-3.5h; the standing time is 7-10h; and the room temperature treatment time is 20-30min.

[0012] Furthermore, the preparation process of the palmitoyl pomegranate peel extract is as follows: Pomegranate peel powder was added to an ethanol-water solution and stirred. The solution was filtered and the filtrate was collected. Palmitic acid was added to the filtrate and heated to react. The solution was concentrated under reduced pressure and then freeze-dried to obtain the palmitoyl pomegranate peel extract.

[0013] Furthermore, the ratio of pomegranate peel powder to ethanol aqueous solution is 1g:(25-35)mL; the volume fraction of the ethanol aqueous solution is 75-85%; the amount of palmitic acid added is 15-25% of the weight of the filtrate; the stirring temperature is 60-70℃ and the time is 40-60min; the heating reaction temperature is 70-80℃ and the time is 3-5h.

[0014] Furthermore, the complex vitamin is composed of vitamin A and vitamin E in a mass ratio of 1:(1-1.5); the amino acid is selected from lysine, methionine, and threonine.

[0015] The preparation method of the above-mentioned *Avicennia marina* extract compound premix includes the following steps: Weigh each ingredient according to the stated weight proportions, stir and mix thoroughly, and it is ready.

[0016] The above-mentioned compound premix of *Vitis pulcherrima* extract is used in the preparation of feed that promotes animal growth.

[0017] The beneficial technical effects of this invention are as follows: 1. The premix of this invention incorporates modified bentonite, which can increase the daily feed intake and daily weight gain of piglets. Specifically, this invention first treats bentonite with zinc chloride, allowing zinc ions to enter the bentonite through ion exchange and improve its pore structure. Treatment of hydroxypropyl guar gum with potassium hydroxide and glycerol deprotonates the hydroxyl groups on its molecular chains, disrupting the hydrogen bond network structure and enhancing reactivity. After freeze-drying, it exhibits a loose, porous structure. The molecular chains of hydroxypropyl guar gum can interpenetrate, entwine, and coat the interlayers and surface of bentonite. Under the action of a cross-linking agent, the interlayer ions of hydroxypropyl guar gum and bentonite coordinate, forming a more stable network. The modified bentonite, utilizing its porous network structure, can better bind mycotoxins in feed or animal intestines. Hydroxypropyl guar gum may enhance its adsorption in the intestines, protecting the intestinal mucosa, promoting the proliferation of beneficial intestinal bacteria, and improving the intestinal microecological balance. Furthermore, its porous adsorption properties can be used as a carrier to increase the residence time of feed in the animal's gastrointestinal tract, promote the digestion and absorption of nutrients, improve feed conversion rate, reduce feed conversion ratio, improve breeding efficiency and reduce costs.

[0018] 2. The premix of this invention also contains palmitoyl pomegranate peel extract, which can increase the levels of IgG, IgA, and IgM in animal serum. Specifically, pomegranate peel extract is rich in polyphenols, which can scavenge free radicals in animals, improve serum antioxidant indicators, and enhance animal immunity. Palmitoyl palmitate oxidase further improves its lipid solubility, allowing it to more easily penetrate the lipid cell membranes of animal cells, thus increasing bioavailability and stability.

[0019] 3. Through the synergistic effect of the above components, this invention comprehensively enhances the intestinal health and immune function of animals, reduces the risk of disease, thereby significantly reducing dependence on antibiotics and hormones, promoting the realization of antibiotic-free farming, and conforming to the industry development trend of green and healthy farming. Attached Figure Description

[0020] Figure 1 SEM image of the modified bentonite prepared in Example 1. Detailed Implementation

[0021] The following is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it should not be construed that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, various simple deductions or substitutions can be made without departing from the concept of the present invention, and all such modifications and substitutions should be considered within the scope of protection of the present invention. Specific conditions not specified in the embodiments are performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, all reagents or instruments used are conventional products obtained through commercial channels.

[0022] The preparation method of the extract from *Agrostis phalloides* of the present invention is as follows: Dissolve Amaryllis in water at a material-to-liquid ratio of 1g:60mL, slowly heat to a gentle boil and beat for 1 hour, change the extraction solvent and beat twice, sonicate at 300W, 40kHz for 50 minutes, filter and freeze dry to obtain Amaryllis extract. The preparation method of reef membrane extract is as follows: The purified reef membrane was dried and pulverized, then passed through a 40-mesh sieve and refluxed with 4 times its weight of water for 2 hours. The reflux liquid was collected and concentrated to 80% of its original volume, then 95% ethanol solution was added and stirred until homogeneous. The mixture was allowed to stand at room temperature for 22 hours, filtered, and the precipitate was collected and dried to obtain the reef membrane extract.

[0023] Preparation Example 1 Preparation Example 1 provides a modified bentonite, prepared by the following method: (1) Bentonite, zinc chloride and deionized water were added to deionized water at a mass ratio of 1:0.06:8. After stirring at room temperature for 2 hours, the mixture was ultrasonically treated for 25 minutes, filtered and washed, and dried at 115°C for 4 hours. Product 1 was collected. (2) According to the mass ratio of potassium hydroxide, hydroxypropyl guar gum, glycerol and deionized water of 1:1:1.5:24, potassium hydroxide was added to deionized water, and then hydroxypropyl guar gum and glycerol were added in sequence. After stirring at room temperature for 50 min, the mixture was freeze-dried and product 2 was collected. (3) According to the mass ratio of product 1, product 2, and deionized water of 1:0.4:22, product 1 was added to deionized water, followed by product 2. After ultrasonic treatment for 3 hours, the mixture was allowed to stand for 8 hours. After centrifugation and washing, the precipitate was collected. According to the volume ratio of precipitate to calcium chloride solution of 1 g:22 mL, the precipitate was added to a calcium chloride solution with a concentration of 11 wt%. The mixture was treated at room temperature for 25 minutes and dried to obtain the modified bentonite. The SEM image of the modified bentonite is shown below. Figure 1 .

[0024] Preparation Example 2 Preparation Example 2 provides a modified bentonite, prepared by the following method: (1) Bentonite, zinc chloride and deionized water were added to deionized water at a mass ratio of 1:0.05:6. After stirring at room temperature for 1 hour, the mixture was ultrasonically treated for 20 minutes, filtered and washed, and dried at 110°C for 5 hours. Product 1 was collected. (2) According to the mass ratio of potassium hydroxide, hydroxypropyl guar gum, glycerol and deionized water 1:0.6:1.2:20, potassium hydroxide was added to deionized water, and then hydroxypropyl guar gum and glycerol were added in sequence. After stirring at room temperature for 40 min, the mixture was freeze-dried and product 2 was collected. (3) According to the mass ratio of product 1, product 2 and deionized water 1:0.3:20, product 1 is added to deionized water, and then product 2 is added. After ultrasonic treatment for 2.5h, the mixture is allowed to stand for 7h. After centrifugation and washing, the precipitate is collected. According to the ratio of precipitate to calcium chloride solution 1g:20mL, the precipitate is added to a 10wt% calcium chloride solution. After treatment at room temperature for 20min, the modified bentonite is obtained after drying.

[0025] Preparation Example 3 Preparation Example 3 provides a modified bentonite, prepared by the following method: (1) Bentonite, zinc chloride and deionized water were added to deionized water at a mass ratio of 1:0.08:10. After stirring at room temperature for 3 hours, the mixture was ultrasonically treated for 30 minutes, filtered and washed, and dried at 120°C for 3 hours. Product 1 was collected. (2) According to the mass ratio of potassium hydroxide, hydroxypropyl guar gum, glycerol and deionized water 1:1.2:2:25, potassium hydroxide was added to deionized water, and then hydroxypropyl guar gum and glycerol were added in sequence. After stirring at room temperature for 60 min, the mixture was freeze-dried and product 2 was collected. (3) According to the mass ratio of product 1, product 2 and deionized water 1:0.5:25, product 1 is added to deionized water, then product 2 is added, ultrasonic treatment is performed for 3.5 h, and then the mixture is allowed to stand for 10 h. After centrifugation and washing, the precipitate is collected. According to the ratio of precipitate to calcium chloride solution 1 g:25 mL, the precipitate is added to a calcium chloride solution with a concentration of 12 wt%, treated at room temperature for 30 min, and dried to obtain the modified bentonite.

[0026] Preparation Example 4 Preparation Example 4 provides a palmitoyl pomegranate peel extract, prepared by the following method: According to the ratio of pomegranate peel powder to ethanol aqueous solution 1g:30mL, pomegranate peel powder was added to 80% ethanol aqueous solution, stirred at 65℃ for 50min, filtered and the filtrate was collected; palmitic acid was added to the filtrate at 20% of the weight of the filtrate, reacted at 75℃ for 4h, concentrated under reduced pressure and freeze-dried to obtain the palmitoyl pomegranate peel extract.

[0027] Preparation Example 5 Preparation Example 5 provides a palmitoyl pomegranate peel extract, prepared by the following method: According to the ratio of pomegranate peel powder to ethanol aqueous solution 1g:25mL, pomegranate peel powder was added to 75% ethanol aqueous solution by volume, stirred at 60℃ for 60min, filtered and the filtrate was collected; palmitic acid was added to the filtrate at 15% of the weight of the filtrate, reacted at 70℃ for 5h, concentrated under reduced pressure and freeze-dried to obtain the palmitoyl pomegranate peel extract.

[0028] Preparation Example 6 Preparation Example 6 provides a palmitoyl pomegranate peel extract, prepared by the following method: According to the ratio of pomegranate peel powder to ethanol aqueous solution 1g:35mL, pomegranate peel powder was added to an 85% ethanol aqueous solution, stirred at 70℃ for 40min, filtered and the filtrate was collected; palmitic acid was added to the filtrate at 15% of the weight of the filtrate, reacted at 80℃ for 3h, concentrated under reduced pressure and freeze-dried to obtain the palmitoyl pomegranate peel extract.

[0029] Example 1 Example 1 provides a compound premix of *Agrostis dentata* extract, comprising the following raw materials in parts by weight: 12 parts of *Agrostis dentata* extract, 3 parts of reef extract, 2 parts of modified bentonite from Preparation Example 1, 3 parts of palmitoyl pomegranate peel extract from Preparation Example 4, 0.4 parts of lysine, and 0.2 parts of compound vitamins; wherein the compound vitamins are composed of vitamin A and vitamin E in a mass ratio of 1:1.2.

[0030] Example 1 also provides a method for preparing the above-mentioned *Ampelopsis thunbergii* extract compound premix, as follows: Weigh each ingredient according to the stated weight proportions, stir and mix thoroughly, and it is ready.

[0031] Example 2 Example 2 provides a compound premix of *Agrostis dentata* extract, comprising the following raw materials in parts by weight: 9 parts *Agrostis dentata* extract, 1 part *Agrostis dentata* extract, 1 part modified bentonite from Preparation Example 2, 2 parts palmitoyl pomegranate peel extract from Preparation Example 5, 0.3 parts methionine, and 0.1 parts compound vitamins; wherein the compound vitamins are composed of vitamin A and vitamin E in a mass ratio of 1:1.

[0032] Example 2 also provides a method for preparing the above-mentioned *Avicennia marina* extract composite premix, as follows: Weigh each ingredient according to the stated weight proportions, stir and mix thoroughly, and it is ready.

[0033] Example 3 Example 3 provides a compound premix of *Agrostis dentata* extract, comprising the following raw materials in parts by weight: 15 parts of *Agrostis dentata* extract, 5 parts of reef extract, 3 parts of modified bentonite from Preparation Example 3, 4 parts of palmitoyl garnet peel extract from Preparation Example 6, 0.5 parts of threonine, and 0.5 parts of compound vitamins; wherein the compound vitamins are composed of vitamin A and vitamin E in a mass ratio of 1:1.5.

[0034] Example 3 also provides a method for preparing the above-mentioned *Avicennia marina* extract compound premix, as follows: Weigh each ingredient according to the stated weight proportions, stir and mix thoroughly, and it is ready.

[0035] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that bentonite and hydroxypropyl guar gum were used instead of the modified bentonite in Preparation Example 1, and the amounts of both were the same as in Preparation Example 1.

[0036] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that pomegranate peel extract was used instead of palmitoyl pomegranate peel extract in Preparation Example 4.

[0037] Experimental Example 1 Sixty healthy Duroc, Landrace, and Large White piglets aged 21 days were selected and randomly divided into the following six groups: control group, Example 1-3 groups, and Comparative Example 1-2 groups, with 10 piglets in each group. The control group was fed a basal diet, while Example 1-3 groups and Comparative Example 1-2 groups were fed a basal diet plus 0.5% of the premixed feed from Example 1-3 groups and Comparative Example 1-2 groups, for 12 consecutive days.

[0038] At the beginning and end of the formal experiment, the fasting weight of piglets in each group was weighed. The amount of feed supplied and the amount of feed remaining were collected and recorded daily. The average daily feed intake (kg / d), average daily weight gain (kg / d), and feed conversion ratio (average daily feed intake / average daily weight gain) were calculated. The results are shown in Table 1.

[0039] After the last feeding, 5 piglets were selected from each group, and 10 mL of blood was collected from the anterior vena cava. The serum was obtained by centrifugation, and the levels of IgG, IgA, and IgM in the serum were then detected using a kit. The results are shown in Table 2.

[0040] Table 1 Table 2 As shown in Table 1, the average daily feed intake and daily weight gain of the Example 1-3 groups were higher than those of the Comparative Example 1-2 groups.

[0041] Compared to Example 1, the premixed feed obtained by replacing modified bentonite with bentonite in Comparative Example 1 resulted in lower daily feed intake and daily weight gain for piglets compared to Example 1. This indicates that modified bentonite can increase the daily feed intake and daily weight gain of piglets. Further analysis reveals that the present invention first treats bentonite with zinc chloride, allowing zinc ions to enter the bentonite through ion exchange, thus improving its pore structure. Treatment of hydroxypropyl guar gum with potassium hydroxide and glycerol deprotonates the hydroxyl groups on its molecular chains, disrupting the hydrogen bond network structure and enhancing reactivity. After freeze-drying, it exhibits a loose, porous structure. The molecular chains of hydroxypropyl guar gum can interpenetrate, entwine, and coat the interlayers and surface of bentonite. Under the action of a crosslinking agent, the interlayer ions of hydroxypropyl guar gum and bentonite coordinate, forming a more stable network. Modified bentonite, with its porous network structure, can better bind mycotoxins in feed or animal intestines. Hydroxypropyl guar gum may enhance its adsorption in the intestines, protect the intestinal mucosa, promote the proliferation of beneficial intestinal bacteria, and improve the balance of the intestinal microecology. Furthermore, its porous adsorption properties allow it to act as a carrier, increasing the residence time of feed in the animal's gastrointestinal tract, promoting the digestion and absorption of nutrients, improving feed conversion ratio, reducing feed conversion ratio, increasing breeding efficiency, and reducing costs.

[0042] As shown in Table 2, the serum levels of IgG, IgA, and IgM in Examples 1-3 were all higher than those in Comparative Examples 1-2.

[0043] Compared to Example 1, in Comparative Example 2, the levels of IgG, IgA, and IgM in the serum of piglets fed with premixed feed obtained by replacing palmitoyl pomegranate peel extract with pomegranate peel extract were lower than in Example 1. This indicates that palmitoyl pomegranate peel extract can increase the levels of IgG, IgA, and IgM in the serum of piglets. Further analysis revealed that pomegranate peel extract is rich in polyphenols, which can scavenge free radicals in animals, improve serum antioxidant indicators, and enhance animal immunity. Palmitoylation with palmitate improves lipid solubility, facilitates penetration through the lipid cell membranes of animal cells, increases bioavailability, and enhances stability.

[0044] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. Based on the present invention, some modifications or substitutions can be made, but these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of protection claimed by the present invention.

Claims

1. A compound premix of *Avicennia marina* extract, characterized in that, The ingredients include the following parts by weight: 9-15 parts of *Ampelopsis grossedentata* extract, 1-5 parts of *Cervi cornuta* extract, 2-4 parts of palmitoyl pomegranate peel extract, 1-3 parts of modified bentonite, 0.3-0.5 parts of amino acids, and 0.1-0.5 parts of complex vitamins.

2. The composite premix of *Ampelopsis grossedentata* extract according to claim 1, characterized in that, The preparation process of the modified bentonite is as follows: (1) Add bentonite and zinc chloride to deionized water, stir at room temperature, and then sonicate. After filtration, washing, and drying, collect product 1. (2) Add potassium hydroxide to deionized water, then add hydroxypropyl guar gum and glycerol in sequence, stir at room temperature and freeze dry, and collect product 2; (3) Add product 1 to deionized water, then add product 2, sonicate and let stand, centrifuge and wash and collect the precipitate; add the precipitate to the crosslinking agent solution for room temperature treatment, and dry to obtain the modified bentonite.

3. The composite premix of *Ampelopsis grossedentata* extract according to claim 2, characterized in that, In step (1), the mass ratio of bentonite, zinc chloride, and deionized water is 1:(0.05-0.08):(6-10); the stirring time is 1-3 hours; the ultrasonic treatment time is 20-30 minutes; and the drying temperature is 110-120°C for 3-5 hours.

4. The composite premix of *Ampelopsis grossedentata* extract according to claim 2, characterized in that, In step (2), the mass ratio of potassium hydroxide, hydroxypropyl guar gum, glycerol, and deionized water is 1:(0.6-1.2):(1.2-2):(20-25); the stirring time is 40-60 min.

5. The composite premix of *Ampelopsis grossedentata* extract according to claim 2, characterized in that, In step (3), the mass ratio of product 1, product 2, and deionized water is 1:(0.3-0.5):(20-25); the volume ratio of the precipitate to the crosslinking agent solution is 1g:(20-25)mL; the crosslinking agent solution is a calcium chloride solution with a concentration of 10-12wt%; the ultrasonic treatment time is 2.5-3.5h; the standing time is 7-10h; and the room temperature treatment time is 20-30min.

6. The composite premix of *Ampelopsis grossedentata* extract according to claim 2, characterized in that, The preparation process of the palmitoyl pomegranate peel extract is as follows: Pomegranate peel powder was added to an ethanol-water solution and stirred. The solution was filtered and the filtrate was collected. Palmitic acid was added to the filtrate and heated to react. The solution was concentrated under reduced pressure and then freeze-dried to obtain the palmitoyl pomegranate peel extract.

7. The composite premix of *Ampelopsis grossedentata* extract according to claim 6, characterized in that, The ratio of pomegranate peel powder to ethanol aqueous solution is 1g:(25-35)mL; the volume fraction of ethanol aqueous solution is 75-85%; the amount of palmitic acid added is 15-25% of the weight of the filtrate; the stirring temperature is 60-70℃ and the time is 40-60min; the heating reaction temperature is 70-80℃ and the time is 3-5h.

8. The compound premix of *Ampelopsis grossedentata* extract according to claim 1, characterized in that, The complex vitamin is composed of vitamin A and vitamin E in a mass ratio of 1:(1-1.5); the amino acid is selected from lysine, methionine, and threonine.

9. A method for preparing a composite premix of *Agrostis phalloides* extract according to any one of claims 1-8, characterized in that, Includes the following steps: Weigh each ingredient according to the stated weight proportions, stir and mix thoroughly, and it is ready.

10. The application of a compound premix of *Avicennia marina* extract according to any one of claims 1-8, characterized in that, It is used in the preparation of feed that promotes animal growth.

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