A lactation promoting composition containing a limonin extract and uses thereof

Abstract

This invention discloses a lactation-promoting composition containing a lime flavonoid extract, comprising, by weight, the following raw material components: 10-50 parts lime flavonoid extract, 5-30 parts synergistic active ingredients, and 20-85 parts excipients; wherein the lime flavonoid extract contains ≥80% total flavonoids (calculated as hesperidin), of which hesperidin ≥40%, neohesperidin ≥15%, naringin ≥10%, and hesperidin ≥5%. Using the lime flavonoid extract as the core, it achieves highly efficient lactation promotion through multi-target synergistic effects. In livestock applications, it can increase milk yield in dairy cows by 15%-28% and milk yield in sows by 20%-38%, while significantly improving milk fat, milk protein, and other milk nutritional indicators.

Description

Technical Field

[0001] This invention belongs to the field of lactation-promoting compositions, and in particular to a lactation-promoting composition containing flavonoid extract of lime and its application. Background Technology

[0002] Lactation is a core physiological process for mammalian reproduction, directly determining the survival rate and growth status of offspring. It is also a key production indicator for livestock in the livestock industry. Furthermore, breastfeeding is the feeding method recommended by the World Health Organization. Postpartum hypogalactia or insufficient milk production is a common postpartum problem for mothers worldwide, directly impacting the rate of exclusive breastfeeding and the long-term health of both mother and child. Currently, existing lactation-promoting technologies and products still have many unresolved shortcomings, specifically as follows: First, hormonal lactation-inducing products pose serious safety risks. While commonly used clinical and aquaculture preparations such as oxytocin, recombinant growth hormone, and estrogen can rapidly increase milk production, they have serious drug residue problems. Their use in livestock farming can affect the food safety of dairy and meat products. Many countries have explicitly banned the illegal use of hormonal lactation-inducing drugs in food animals.

[0003] Secondly, traditional Chinese medicine galactagogue compositions suffer from uncontrollable quality and unstable efficacy. Classic galactagogue formulas such as Yongquan San and Tongru Dan are mostly composed of multiple Chinese herbs, including Wangbuliuxing, Tongcao, Danggui, and Huangqi. While they have certain galactagogue effects and relatively high safety, they have three major drawbacks: First, their complex composition lacks clear active ingredient and content indicators, resulting in significant batch-to-batch quality variations and making standardized industrial production difficult. Second, they have a slow onset of action, typically requiring 7-14 days of continuous use to show results, failing to quickly improve acute hypogalactia. Third, their mechanism of action is unclear, largely based on traditional Chinese medicine theories of "tonifying qi and blood, promoting menstruation and lactation," lacking modern molecular biological mechanism verification, making it difficult to gain international market acceptance.

[0004] Third, existing lactation-promoting products based on natural plant extracts have low activity and limited effects. Currently, a small number of lactation-promoting products based on phytoestrogens such as soy isoflavones and pueraria isoflavones have been developed. However, these isoflavone components have weak binding ability to estrogen receptors, low bioavailability, and limited lactation-promoting effects, typically only increasing milk production by 5%-10%, which cannot meet the application needs of high-demand scenarios. At the same time, most plant extracts have poor palatability and strong odor, resulting in low feed intake in livestock farming.

[0005] Bitter orange (*Citrus aurantium* L.) is a plant of the genus Citrus in the Rutaceae family. Its dried immature fruit is the traditional Chinese medicine Fructus Aurantii Immaturus, and its dried nearly mature pericarp is Fructus Aurantii. It has a medicinal history of more than 2,000 years in China. Existing research shows that the core active components in bitter orange are flavonoids, including hesperidin, neohesperidin, naringin, hesperetin, nobiletin, etc. The current research and application of it mainly focus on directions such as antioxidant, anti-inflammatory, weight loss, regulation of intestinal flora, and improvement of cardiovascular function. There are no relevant reports at home and abroad on the lactation-promoting activity, mechanism of action of bitter orange flavonoid extract, and lactation-promoting composition based on this extract, presenting a significant technical gap.

[0006] Based on this, there is an urgent need in this field to develop a lactation-promoting composition with clear ingredients, controllable quality, significant lactation-promoting effect, safe without side effects, and wide application range, to fill the application gap of bitter orange flavonoids in the field of lactation promotion and solve the core industry pain points of existing lactation-promoting products. Summary of the Invention

[0007] The primary object of the present invention is to provide a lactation-promoting composition containing bitter orange flavonoid extract. This composition uses the standardized prepared bitter orange flavonoid extract as the core active ingredient, achieving a high-efficiency lactation-promoting effect through multi-target synergistic action, and at the same time having the advantages of clear ingredients, controllable quality, safe without residues, and good palatability. The second object of the present invention is to provide a preparation method for the lactation-promoting composition containing bitter orange flavonoid extract. This method has stable process, high extraction efficiency, and complete retention of active ingredients, and is suitable for large-scale industrial production. The third object of the present invention is to provide the application of the lactation-promoting composition containing bitter orange flavonoid extract, including in lactation-promoting feed additives and veterinary drug preparations for livestock and poultry farming, providing a safe and efficient solution for improving the lactation performance and postpartum lactation induction of livestock and poultry farming.

[0008] The present invention discloses a lactation-promoting composition containing bitter orange flavonoid extract. Calculated by weight份, it includes the following raw material components: 10 - 50 parts of bitter orange flavonoid extract, 5 - 30 parts of synergistic active ingredient, and 20 - 85 parts of excipient; in the bitter orange flavonoid extract, the total flavonoid content calculated by hesperidin is ≥80%, among which hesperidin ≥40%, neohesperidin ≥15%, naringin ≥10%, and hesperetin ≥5%.

[0009] As a further improvement of the above technical solution: The preparation method of the bitter orange flavonoid extract includes the following steps: Raw material pretreatment: Take the dried mature fruit of bitter orange, crush it and pass through a 40-mesh sieve to obtain bitter orange raw material powder; Degreasing pretreatment: The bitter orange raw material powder is refluxed with petroleum ether at 60°C for 2 times, each time for 2 hours, filtered, and the filter residue is dried to remove petroleum ether to obtain degreased bitter orange powder; Ultra-high pressure assisted ethanol extraction: Add 70% ethanol aqueous solution by volume to defatted lime powder, with a material-liquid ratio of 1:15. After mixing evenly, transfer to an ultra-high pressure extraction device. Set the ultra-high pressure to 300 MPa and the extraction temperature to 50℃. Extract 3 times, 40 min each time. After extraction, collect and combine the extracts, and recover the ethanol under reduced pressure to obtain flavonoid concentrate. Macroporous resin purification: Dilute the flavonoid concentrate to a total flavonoid concentration of 2 mg / mL, load the sample onto an AB-8 macroporous resin column at a flow rate of 2 BV / h and a sample volume of 4 BV; wash with 5 BV of deionized water to remove impurities, then elute with 6 BV of 70% ethanol aqueous solution and collect the ethanol eluent. Concentration and drying: Ethanol eluent was recovered under reduced pressure, and the solution was freeze-dried under vacuum to obtain the flavonoid extract of orange.

[0010] The synergistic active ingredients are one or more of the following: Wangbuliuxing flavonoids, Angelica sinensis polysaccharides, Codonopsis pilosula polysaccharides, Astragaloside A, Astragalus polysaccharides, vitamin E, folic acid, β-carotene, and probiotic freeze-dried powder.

[0011] The composition is available in the form of a livestock feed premix, soluble powder, oral liquid, or granules.

[0012] The excipients for the animal husbandry formulation are one or more of the following: corn starch, wheat bran, skim milk powder, limestone powder, maltodextrin, Tween-80, potassium sorbate, white sugar, and sodium carboxymethyl cellulose.

[0013] This invention discloses a method for preparing a lactation-promoting composition containing flavonoid extract of lime, which includes the following steps when the composition is in solid dosage form: Raw material pretreatment: Pulverize the lime flavonoid extract, synergistic active ingredients and excipients separately, pass them through an 80-100 mesh sieve, and weigh them according to the formula weight parts for later use; Mixing: Place the weighed raw materials into a three-dimensional motion mixer and mix for 20-40 minutes until the mixing uniformity RSD ≤ 5% to obtain a mixed powder; Forming and Packaging: Depending on the dosage form requirements, the mixed powder is directly packaged, or it is formed after granulation, drying, sizing, filling / tableting, and then packaged after passing quality inspection to obtain the finished product.

[0014] This invention discloses a method for preparing a lactation-promoting composition containing flavonoid extract of lime, wherein when the composition is in the form of an oral liquid, the method includes the following steps: Raw material pretreatment: Weigh each raw material according to the formula weight parts and set aside; Solution preparation: Take 80% of the prescribed amount of deionized water, add solubilizer, stir evenly, then add lime flavonoid extract and synergistic active ingredients, stir until completely dissolved; then add the remaining excipients, stir until completely dissolved, and add deionized water to the total volume to obtain the drug solution; Filtration, sterilization and packaging: The liquid medicine is filtered through a 0.45μm microporous membrane, filled, capped and sterilized. After passing quality inspection, it is packaged to obtain the finished product.

[0015] As a further improvement to the above technical solution: This invention discloses the application of a lactation-promoting composition containing flavonoid extract of lime in the preparation of lactation-promoting products. The lactation-promoting products are lactation-promoting feed additives or veterinary drug preparations for livestock breeding, including dairy cows, sows, meat sheep, and rabbits.

[0016] Compared with the prior art, the beneficial effects of the present invention are: The composition of this invention uses lime flavonoid extract as its core, achieving highly efficient lactation promotion through multi-target synergistic effects. In livestock applications, it can increase milk yield in dairy cows by 15%-28% and milk yield in sows by 20%-38%, while significantly improving milk fat, milk protein, and other milk nutritional indicators.

[0017] This invention achieves precise quantification of total flavonoids and core monomer components in lime flavonoid extract through standardized extraction and purification processes. The composition formula is fixed with small batch-to-batch differences, solving the pain points of complex composition and uncontrollable quality of traditional Chinese medicine compositions. The preparation process is simple and stable, the raw materials are widely available (my country has a large planting area of ​​lime and high yield), and the production cost is controllable, making it suitable for large-scale industrial production.

[0018] All active ingredients in the composition of this invention are derived from natural edible plants and contain no hormones. Acute toxicity and subchronic toxicity tests have verified that it is practically non-toxic and has no teratogenic, mutagenic, or carcinogenic effects. There is no withdrawal period in livestock applications, and no drug residues will be produced in animal-derived foods, which meets national food safety standards.

[0019] The flavonoid extract of lime of the present invention has no obvious odor and can be prepared into various dosage forms according to the application scenario. It is suitable for the breeding and production of lactating economic livestock such as dairy cows, sows, sheep, and rabbits.

[0020] This invention is the first to discover and verify the lactation-promoting activity of lime flavonoid extract, clarify its molecular mechanism of action, and fill the technological gap in related fields at home and abroad. It breaks through the limitation of single-target action of existing lactation-promoting products, and achieves stable and long-lasting effects through the synergistic action of four major pathways: endocrine regulation, microcirculation improvement, antioxidant and anti-inflammatory effects, and intestinal flora regulation. The effect is stable and long-lasting and not prone to rebound. Detailed Implementation

[0021] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below in conjunction with specific embodiments.

[0022] In the description of this invention, it should be noted that the terms "upper," "lower," "inner," "outer," "front end," "rear end," "both ends," "one end," and "the other end," etc., indicate the orientation or positional relationship based on the shown orientation or positional relationship, and are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0023] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," "connected," etc., should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0024] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0025] The sour orange flavonoid extract of this invention is prepared from dried, ripe sour orange fruit through an ultrasonic-assisted ethanol extraction-macroporous resin purification process. The total flavonoid content in the extract is ≥80% (calculated as hesperidin, detected by ultraviolet spectrophotometry). The proportions of each core active ingredient are: hesperidin ≥40%, neohesperidin ≥15%, naringin ≥10%, and hesperidin ≥5%, thus achieving standardization and quality control of the active ingredients.

[0026] The preparation method of the lime flavonoid extract specifically includes the following steps: 1. Raw material pretreatment: Take dried and ripe sour oranges, remove moldy and impurity raw materials, crush them with a universal pulverizer, pass them through a 40-mesh standard sieve to obtain sour orange raw material powder, and store them in a sealed and light-proof container for later use. 2. Degreasing pretreatment: Take lime raw material powder, add petroleum ether (boiling range 60-90℃) at a material-to-liquid ratio of 1:5 (g:mL), and degrease twice by reflux in a 60℃ water bath for 2 hours each time. After degreasing, filter with a Buchner funnel, place the filter residue in a fume hood to evaporate the petroleum ether, and obtain degreased lime powder. 3. Ultra-high pressure assisted ethanol extraction: Add 70% (v / v) ethanol aqueous solution to defatted lime powder at a material-to-liquid ratio of 1:15. After mixing thoroughly, transfer the mixture to an ultra-high pressure extraction device. Set the ultra-high pressure to 300 MPa and the extraction temperature to 50℃. Extract three times, 40 min each time. After extraction, collect and combine the extracts, and recover the ethanol under reduced pressure to obtain a flavonoid concentrate. After extraction, combine the three extracts and recover the ethanol under reduced pressure using a vacuum rotary evaporator at 55℃ and -0.09 MPa until the extract has no alcohol odor, thus obtaining a flavonoid concentrate. 4. Macroporous resin purification: Dilute the flavonoid concentrate with deionized water to a total flavonoid concentration of 2 mg / mL, stir well, and then load the solution onto a pretreated AB-8 macroporous resin column. Set the loading flow rate to 2 BV / h and the loading volume to 4 times the column volume (BV). After loading, elute with 5 BV of deionized water at a flow rate of 3 BV / h to remove impurities such as water-soluble polysaccharides and proteins. After impurity removal, elute with 6 BV of 70% ethanol aqueous solution at a flow rate of 2 BV / h and collect the ethanol eluent. 5. Concentration and drying: The collected ethanol eluent was placed in a vacuum rotary evaporator and the ethanol was recovered under reduced pressure at 55℃ and -0.09MPa to obtain a purified and concentrated flavonoid solution. The purified and concentrated solution was placed in a vacuum freeze dryer, with a pre-freezing temperature of -40℃, a pre-freezing time of 12h, a cold trap temperature of -55℃, a vacuum degree of 10Pa, and a drying time of 24h to obtain a light yellow powdery orange flavonoid extract, which was sealed and stored in a light-proof environment at 4℃.

[0027] The lactation-promoting composition of the present invention comprises, by weight, the following raw material components: 10-50 parts of lime flavonoid extract, 5-30 parts of synergistic active ingredients, and 20-85 parts of excipients. Furthermore, the synergistic active ingredients are natural active ingredients that have synergistic effects in promoting lactation, replenishing qi and blood, and improving nutrient absorption, including but not limited to Wangbuliuxing flavonoids, Angelica sinensis polysaccharides, Codonopsis pilosula polysaccharides, Astragaloside A, Astragalus polysaccharides, vitamin E, folic acid, β-carotene, and one or more combinations of probiotic freeze-dried powder (one or more of Bifidobacterium animalis, Lactobacillus rhamnosus, and Lactobacillus plantarum).

[0028] Furthermore, the excipients are selected according to the application scenario and dosage form of the composition, wherein: the excipients for livestock feed additives / veterinary drug preparations include one or more of the following: corn starch, wheat bran, skim milk powder, limestone powder, maltodextrin, Tween-80, potassium sorbate, white sugar, and sodium carboxymethyl cellulose.

[0029] Furthermore, the lactation-promoting composition of the present invention can be prepared into various dosage forms, including but not limited to: livestock feed premix, soluble powder, oral liquid, and granules.

[0030] The preparation method of the lactation-promoting composition of the present invention includes the following core processes depending on the dosage form, and all processes are carried out in a clean room that meets GMP requirements: 1. Preparation methods for solid dosage forms (premixes, soluble powders, capsules, tablets, granules): (1) Raw material pretreatment: Crush the lime flavonoid extract, synergistic active ingredients and excipients separately, pass them through an 80-100 mesh standard sieve, and weigh them according to the formula weight parts for later use; (2) Mixing: Place the weighed raw materials in a three-dimensional motion mixer and mix for 20-40 minutes until the mixing uniformity RSD ≤ 5% to obtain mixed powder; (3) Forming: According to the dosage form requirements, the mixed powder is directly packaged to obtain premix / soluble powder; or granules are obtained after granulation, drying and sizing; or capsules are obtained by filling capsules; or tablets are obtained by adding lubricant and then compressing. (4) Quality inspection and packaging: The content, moisture and microbial limit of the molded products are tested. After passing the test, the products are packaged into inner and outer packaging and stored in the warehouse.

[0031] 2. Preparation method of liquid dosage form (oral liquid): (1) Raw material pretreatment: Weigh the lime flavonoid extract, synergistic active ingredients and excipients according to the formula weight parts and set aside; (2) Solution preparation: Take 80% of the prescription amount of deionized water, add solubilizer Tween-80, stir evenly, add lime flavonoid extract and synergistic active ingredients, stir at 300 r / min until completely dissolved; then add sweetener, preservative and other excipients, stir until completely dissolved, add deionized water to the total volume to obtain the drug solution; (3) Filtration and sterilization: The drug solution is filtered through a 0.45μm microporous membrane, the filtrate is filled into oral liquid bottles, the caps are crimped and sealed, and the oral liquid product is sterilized by flowing steam at 105℃ for 30 minutes to obtain the finished oral liquid product; (4) Quality inspection and packaging: The content, pH value and microbial limit of the finished product are tested. After passing the test, the product is packaged and stored in the warehouse.

[0032] (iv) Mechanism of Action of the Lactation-Promoting Composition The lactation-promoting composition containing lime flavonoid extract described in this invention achieves a highly efficient lactation-promoting effect through the synergistic action of multiple targets and pathways. The core mechanism of action is as follows: 1. Regulating the hypothalamus-pituitary-mammary axis and upregulating the expression of lactation-related hormones and receptors: The active ingredients such as hesperidin and hesperidin in lime flavonoids can specifically bind to estrogen receptors ERα and ERβ in mammary tissue, activate the hypothalamus-pituitary-gonadal axis, and promote the pituitary gland to synthesize and secrete prolactin (PRL) and growth hormone (GH). At the same time, it can significantly upregulate the mRNA and protein expression levels of prolactin receptor (PRLR) in mammary tissue, improve the sensitivity of mammary cells to prolactin, promote the proliferation and differentiation of mammary acinar epithelial cells, increase the number of lactating cells, and fundamentally enhance the lactation capacity of the mammary gland.

[0033] 2. Improves mammary microcirculation, providing sufficient material basis for lactation: Lime flavonoid extract can inhibit angiotensin-converting enzyme activity, dilate capillaries in mammary tissue, reduce blood viscosity, and improve blood perfusion in mammary tissue; at the same time, it can upregulate the expression of vascular endothelial growth factor (VEGF), promote angiogenesis in mammary tissue, and improve the efficiency of transport of nutrients and oxygen to mammary lactogenic cells, providing sufficient nutrition and energy support for milk synthesis.

[0034] 3. Antioxidant and anti-inflammatory effects, protecting normal mammary gland function: Lactating mammals are prone to mammary gland inflammation (such as mastitis in dairy cows and mammary edema in postpartum women) due to oxidative stress and bacterial infection, leading to impaired lactation function. Lime flavonoid extract can eliminate excess oxygen free radicals in the body, increase the activity of antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and reduce the content of malondialdehyde (MDA) oxidative damage products. Simultaneously, it can significantly inhibit the expression of pro-inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β), alleviating the inflammatory response in mammary tissue, protecting mammary lactating cells from oxidative and inflammatory damage, and maintaining normal mammary gland lactation function.

[0035] 4. Regulating gut microbiota and controlling lactation through the gut-mammary axis: As a natural prebiotic, lime flavonoid extract can selectively promote the proliferation of beneficial bacteria such as Bifidobacteria and Lactobacillus in the gut, inhibit the growth of harmful bacteria such as Escherichia coli and Salmonella, regulate the balance of gut microbiota, improve intestinal barrier function, and increase the body's absorption rate of lactation-related nutrients such as protein, amino acids, and vitamins. At the same time, through the gut-mammary axis, it can transport active substances such as short-chain fatty acids produced by gut microbiota metabolism to mammary tissue, regulate the immune function of mammary glands and milk synthesis capacity, and achieve an indirect lactation-promoting effect.

[0036] 5. Synergistic effect of active ingredients: The flavonoid glycosides of Vaccaria segetalis in the composition can synergistically promote prolactin secretion and unclog mammary ducts; Angelica polysaccharides and Astragalus polysaccharides can replenish qi and blood, and improve the qi and blood deficiency of lactating animals / parturients; Vitamin E and folic acid can supplement the essential nutrients during lactation, improve the quality of milk, and form a multi-target synergistic effect with the flavonoid extract of Citrus aurantium, further enhancing the lactation-promoting effect. Example 1

[0037] Preparation of lime flavonoid extract. The specific steps for preparing lime flavonoid extract in this embodiment are as follows: 1. Raw material pretreatment: Take ripe sour oranges from Lianyuan, Hunan, remove moldy and impurity raw materials, crush them with a universal pulverizer, pass them through a 40-mesh standard sieve to obtain sour orange raw material powder, and seal and store them in the dark for later use. 2. Degreasing pretreatment: Take 10 kg of lime raw material powder, add 50 L of petroleum ether (boiling range 60-90℃), and degrease twice by reflux in a 60℃ water bath for 2 hours each time. After degreasing, filter with a Buchner funnel, and place the filter residue in a fume hood to evaporate the petroleum ether, to obtain 9.2 kg of degreased lime powder. 3. Ultra-high pressure assisted ethanol extraction: Add 138L of 70% ethanol aqueous solution to 9.2kg defatted lime powder, with a material-to-liquid ratio of 1:15 (g:mL). After thorough mixing, transfer to a food-grade ultra-high pressure extraction vessel, seal, and pressurize to 300MPa at a rate of 20-30MPa / s. Control the extraction temperature at 50℃ and maintain the pressure for 6min. Then, depressurize at a rate of 10-15MPa / s. Repeat the above extraction operation twice, and combine the three extracts (Note: the first extraction is the main extraction, the second extraction is the residue re-extraction, and the total extraction rate is ≥96%). Use a vacuum rotary evaporator at 55℃ and -0.09MPa to recover ethanol under reduced pressure until the extract has no alcohol odor, yielding 12L of flavonoid concentrate.

[0038] 4. Macroporous resin purification: Dilute the flavonoid concentrate with deionized water to a total flavonoid concentration of 2 mg / mL, with a final volume of 45 L. After stirring evenly, load the solution onto a pretreated AB-8 macroporous resin column (resin volume 10 L, column diameter to height ratio 1:8). Set the loading flow rate to 20 L / h and the loading volume to 40 L. After loading, elute with 50 L of deionized water at a flow rate of 30 L / h to remove impurities such as water-soluble polysaccharides and proteins. After impurity removal, elute with 60 L of 70% ethanol aqueous solution at a flow rate of 20 L / h and collect the ethanol eluent. 5. Concentration and drying: The collected ethanol eluent was placed in a vacuum rotary evaporator and the ethanol was recovered under reduced pressure at 55℃ and -0.09MPa to obtain 3.5L of purified flavonoid concentrate. The purified concentrate was placed in a vacuum freeze dryer, with a pre-freezing temperature of -40℃, a pre-freezing time of 12h, a cold trap temperature of -55℃, a vacuum degree of 10Pa, and a drying time of 24h to obtain 862g of light yellow powdered orange flavonoid extract. It was sealed and stored in a light-proof environment at 4℃.

[0039] Content detection: The total flavonoid content was determined by ultraviolet spectrophotometry, using hesperidin as a reference standard. The total flavonoid content in the lime flavonoid extract prepared in this example was found to be 85.2%. The content of each monomer component was determined by high performance liquid chromatography (HPLC). The results were: hesperidin 45.6%, neohesperidin 18.3%, naringin 12.1%, and hesperidin 5.8%, which met the quality requirements of this invention. Example 2

[0040] Preparation of a premixed lactation-promoting feed additive for dairy cows. This embodiment prepares a premixed lactation-promoting feed additive for dairy cows. The formula, by weight, is as follows: 30 parts of lime flavonoid extract prepared in Example 1, 10 parts of Wangbuliuxing flavonoid glycosides, 8 parts of Angelica sinensis polysaccharide, 2 parts of astragaloside A, 3 parts of vitamin E, 0.5 parts of folic acid, and 46.5 parts of corn starch. The preparation method is as follows: 1. Raw material pretreatment: Crush each raw material, pass it through an 80-mesh standard sieve, and weigh it according to the formula weight parts for later use; 2. Mixing: Place the weighed raw materials into a three-dimensional motion mixer and mix for 30 minutes. Take a sample to test the mixing uniformity. RSD ≤ 5% is considered qualified, and the mixed powder is obtained. 3. Packaging and dispensing: The mixed powder is dispensed into 1kg / bag aluminum foil bags, sealed, and packaged to obtain the finished product of the lactation-promoting feed additive premix for dairy cows. Example 3

[0041] Preparation of an oral lactation-promoting solution for sows: This embodiment describes the preparation of an oral lactation-promoting solution for sows. The formula, by weight, is as follows: 20 parts of lime flavonoid extract prepared in Example 1, 8 parts of Wangbuliuxing flavonoid glycosides, 10 parts of Codonopsis pilosula polysaccharide, 2 parts of vitamin E, 0.2 parts of folic acid, 5 parts of Tween-80, 0.3 parts of potassium sorbate, 10 parts of white sugar, and 44.5 parts of deionized water. The preparation method is as follows: 1. Raw material pretreatment: Weigh each raw material according to the formula weight parts and set aside; 2. Solution preparation: Take 80% of the prescribed amount of deionized water, add Tween-80, stir at 300 r / min until well mixed, then add lime flavonoid extract, Wangbuliuxing flavonoid glycosides, and Codonopsis polysaccharide, and stir until completely dissolved; then add vitamin E, folic acid, white sugar, and potassium sorbate, and stir until completely dissolved, then add deionized water to the total volume to obtain the medicinal solution; 3. Filtration and sterilization: The drug solution is filtered through a 0.45μm microporous membrane. The filtrate is filled into oral liquid bottles at 20mL / bottle, capped and sealed, and sterilized by flowing steam at 105℃ for 30min to obtain the finished oral liquid product; 4. Quality inspection and packaging: The content, pH value and microbial limit of the finished product are tested. After passing the test, the product is packaged and stored in the warehouse. Example 4

[0042] Preparation of a lactation-promoting soluble powder for mutton sheep: This embodiment prepares a lactation-promoting soluble powder for mutton sheep. The formula, by weight, is as follows: 15 parts of lime flavonoid extract prepared in Example 1, 5 parts of Wangbuliuxing flavonoid glycosides, 10 parts of Astragalus polysaccharide, 2 parts of vitamin E, and 68 parts of maltodextrin. The preparation method is as follows: 1. Raw material pretreatment: Crush each raw material, pass it through an 80-mesh standard sieve, and weigh it according to the formula weight parts for later use; 2. Mixing: Place the weighed raw materials into a three-dimensional motion mixer and mix for 30 minutes until the mixing uniformity RSD ≤ 5%, to obtain a mixed powder; 3. Packaging and dispensing: The mixed powder is dispensed into 500g bags in aluminum foil bags, sealed, and then packaged to obtain the finished product of lactation-promoting soluble powder for mutton sheep.

[0043] Comparative Example 1 Blank control composition: This comparative example prepared a blank control composition by replacing the lime flavonoid extract in Example 2 with an equal amount of corn starch. The rest of the formulation and preparation method were completely consistent with Example 2.

[0044] Comparative Example 2 Soy isoflavone control composition: This comparative example prepared a soy isoflavone control composition by replacing the lime flavonoid extract in Example 2 with an equal amount of soy isoflavones (total isoflavone content 80%). The rest of the formulation and preparation method are completely consistent with Example 2.

[0045] Experimental Example 1 Acute toxicity test of lime flavonoid extract: This experimental case verifies the acute toxicity of lime flavonoid extract. The experiment was conducted in accordance with the "GB15193.3-2014 National Food Safety Standard Acute Oral Toxicity Test".

[0046] 1. Experimental animals: 40 SPF-grade Kunming mice, half male and half female, weighing 18-22g, were provided by Hunan Slack Jingda Experimental Animal Co., Ltd., animal license number: SCXK (Xiang) 2020-0004; the mice were acclimatized for 3 days before the experiment, with free access to food and water, an ambient temperature of 22±2℃, a relative humidity of 50±5%, and a 12-hour light-dark cycle.

[0047] 2. Experimental Methods: The maximum tolerated dose (MTD) method was used. The lime flavonoid extract prepared in Example 1 was prepared into a suspension of 200 mg / mL with 0.5% sodium carboxymethyl cellulose. Before the experiment, mice were fasted for 16 hours but allowed free access to water. Mice were gavaged at a dose of 20 mL / kg body weight three times within 24 hours, with a cumulative dose of 12 g / kg body weight. After gavage, mice were observed for 14 consecutive days. The general condition, behavior, diet, defecation, and mortality of mice were recorded daily, and their body weight was measured weekly. After the 14-day experiment, the mice were euthanized, and the gross pathological changes of major organs such as the heart, liver, spleen, lungs, kidneys, stomach, and intestines were observed.

[0048] 3. Experimental results: (1) During the 14-day observation period, all 40 mice survived without any deaths; (2) After gavage, the mice did not exhibit any abnormal behavior, had normal eating and defecation, and showed no symptoms of poisoning such as diarrhea, vomiting, convulsions, or drowsiness. Their fur was smooth and their mental state was good; (3) The mice gained weight normally, and there was no significant difference in weight changes between male and female mice and the blank control group (P>0.05); (4) The anatomical observation results showed that the main organs of the mice, such as the heart, liver, spleen, lungs, and kidneys, were normal in morphology, and there were no visible pathological changes such as swelling, congestion, necrosis, or atrophy.

[0049] 4. Experimental conclusion: The maximum tolerated acute oral dose (MTD) of the lime flavonoid extract prepared in this invention in mice is >12 g / kg body weight. According to the acute toxicity classification standard, it belongs to the **practically non-toxic class**, indicating extremely high safety.

[0050] Experimental Example 2 The subchronic toxicity test of lime flavonoid extract was conducted to verify the safety of long-term use of lime flavonoid extract. The test was carried out in accordance with the "GB15193.13-2015 National Food Safety Standard 90-day oral toxicity test".

[0051] 1. Experimental animals: 80 SPF-grade SD rats, half male and half female, weighing 80-100g, were provided by Hunan Slack Jingda Experimental Animal Co., Ltd., animal license number: SCXK (Xiang) 2020-0004; the rats were acclimatized for 3 days before the experiment, with free access to food and water, an ambient temperature of 22±2℃, a relative humidity of 50±5%, and a 12h light-dark cycle.

[0052] 2. Experimental grouping: Rats were randomly divided into 4 groups of 20 each, half male and half female, namely: (1) Blank control group: gavage with an equal volume of 0.5% sodium carboxymethyl cellulose solution; (2) Low-dose group: gavage with orange flavonoid extract at a dose of 100 mg / kg body weight / day; (3) Medium-dose group: gavage with orange flavonoid extract at a dose of 500 mg / kg body weight / day; (4) High-dose group: gavage with orange flavonoid extract at a dose of 2000 mg / kg body weight / day; Each group of rats was gavage once a day for 90 consecutive days, during which they were allowed free access to basic feed and water.

[0053] 3. Detection indicators: (1) General observation: Observe the general condition, behavior, diet, defecation, poisoning symptoms and death of rats every day, and weigh and eat ingestion every week; (2) Blood routine and blood biochemical test: On the 90th day of the experiment, the rats were fasted for 12 hours, blood was taken from the orbital cavity, and blood routine indicators (white blood cells, red blood cells, hemoglobin, platelets, etc.) and blood biochemical indicators (alanine aminotransferase, aspartate aminotransferase, creatinine, urea nitrogen, blood glucose, total cholesterol, triglycerides, etc.) were tested; (3) Organ coefficient and pathological histological examination: After blood collection, the rats were euthanized, and organs such as heart, liver, spleen, lung, kidney, brain, uterus, and ovary were dissected, weighed wet weight, and organ coefficient was calculated; each organ was fixed with 4% paraformaldehyde, embedded in paraffin, sectioned, stained with HE, and the pathological histological changes were observed under an optical microscope.

[0054] 4. Experimental results: (1) General condition: During the 90-day gavage period, there were no deaths or abnormal behaviors in the rats in each group. Their diet and defecation were normal, their fur was smooth, and their mental state was good. There was no significant difference in weight gain and food intake between the rats in each dose group and the blank control group (P>0.05). (2) Blood routine and blood biochemistry: The blood routine and blood biochemistry indicators of the rats in each dose group were all within the normal physiological range and there was no significant difference between them and the blank control group (P>0.05), indicating that long-term use of lime flavonoid extract had no adverse effects on the hematopoietic function and liver and kidney function of the rats. (3) Organ coefficient: There was no significant difference in the coefficient of the main organs of the rats in each dose group and the blank control group (P>0.05), and there were no abnormalities such as organ enlargement or atrophy. (4) Pathological histological examination: Under the light microscope, the tissue structure of the main organs such as heart, liver, spleen, lung, and kidney of the rats in each dose group was normal, and there was no pathological damage such as inflammation, degeneration, necrosis, or hyperplasia. There was no significant difference between them and the blank control group.

[0055] 5. Experimental conclusion: After continuous oral administration of up to 2000 mg / kg body weight of lime flavonoid extract to rats for 90 days, no obvious toxic reactions or target organ damage were found, indicating that the lime flavonoid extract of the present invention is safe and reliable for long-term use and has no cumulative toxicity.

[0056] Experimental Example 3 Experiment on the effect of the composition on the lactation performance of dairy cows: This experiment verifies the effect of the lactation-promoting composition for dairy cows prepared in Example 2 on the lactation performance of dairy cows.

[0057] 1. Experimental animals and grouping: Forty Holstein cows from a large-scale dairy farm in Hunan Province were selected. They were in their 2nd or 3rd parity, in the middle of lactation (90-120 days of lactation), with similar daily milk production, healthy and free from disease, and had no history of mastitis. The cows were randomly divided into 4 groups of 10 each: (1) Blank control group: fed a basic diet without any lactation-promoting additives; (2) Comparative example 1 group: fed a basic diet + the blank control premix of comparative example 1, with an addition amount of 100g / head / day; (3) Comparative example 2 group: fed a basic diet + the soybean isoflavone control premix of comparative example 2, with an addition amount of 100g / head / day; (4) Example 2 group: fed a basic diet + the lactation-promoting premix of example 2, with an addition amount of 100g / head / day. During the experiment, the basic diet and feeding management conditions of each group of cows were completely consistent. They had free access to water and were milked twice a day, morning and evening.

[0058] 2. Experimental period: 7 days for the preliminary test and 60 days for the formal test.

[0059] 3. Detection indicators: (1) Milk production: Record the milk production of each cow in the morning and evening every day, calculate the average daily milk production, and statistically analyze the changes in milk production throughout the experiment; (2) Milk quality indicators: On day 0, day 15, day 30, day 45 and day 60 of the experiment, collect 50 mL of mixed milk samples from each cow in the morning and evening, and use a dairy analyzer to detect the milk fat percentage, milk protein percentage, lactose percentage and non-fat solids content; (3) Milk somatic cell count: On day 60 of the experiment, collect milk samples, use a somatic cell counter to detect the somatic cell count in the milk, and assess the health status of the mammary glands; (4) Serum hormone indicators: On day 60 of the experiment, collect 10 mL of tail vein blood from cows in a fasting state, separate the serum, and use enzyme-linked immunosorbent assay (ELISA) to detect the content of prolactin (PRL), growth hormone (GH) and estradiol (E2) in the serum.

[0060] 4. Statistical analysis of experimental data: SPSS 26.0 software was used for statistical analysis. Quantitative data were expressed as mean ± standard deviation. One-way ANOVA was used for comparison between groups. P < 0.05 was considered statistically significant, and P < 0.01 was considered highly statistically significant.

[0061] 5. Experimental results: (1) Effect on milk production of dairy cows: Before the start of the experiment (day 0), there was no significant difference in the average daily milk production of dairy cows in each group (P>0.05); on day 60 of the experiment, the average daily milk production of dairy cows in each group is shown in Table 1.

[0062] Table 1. Comparison of average daily milk yield of dairy cows in each group on day 60 of the experiment. As shown in Table 1, the average daily milk yield of dairy cows in Example 2 group increased by 28.25% compared with the blank control group, which was significantly higher than that of the blank control group and Comparative Example 1 (P<0.01), and significantly better than the soybean isoflavone control group of Comparative Example 2 (P<0.05). This indicates that the composition of the present invention can significantly increase the milk yield of dairy cows, and the effect is far superior to existing similar products.

[0063] (2) Effect on milk quality of dairy cows: On the 60th day of the experiment, the milk quality test results of each group of dairy cows are shown in Table 2.

[0064] Table 2 Comparison of milk quality indicators among different groups As shown in Table 2, the milk fat percentage, milk protein percentage, lactose percentage, and non-fat solids content of the Example 2 group were significantly higher than those of the blank control group (P < 0.01) and significantly higher than those of the Comparative Example 2 group (P < 0.05), indicating that the composition of the present invention can not only increase milk production but also significantly improve the nutritional quality of milk. (3) Effects on milk somatic cell count and serum hormones: On day 60 of the experiment, the milk somatic cell count of the Example 2 group was 125,000 ± 12,000 / mL, which was significantly lower than that of the blank control group (356,000 ± 28,000 / mL) (P < 0.01), indicating that the composition of the present invention can significantly improve the health of the mammary glands of dairy cows and reduce the risk of mastitis. Serum hormone test results showed that the serum prolactin level in the Example 2 group was 18.6±1.5 ng / mL, which was 82.35% higher than that in the blank control group (10.2±0.8 ng / mL). The levels of growth hormone and estradiol were also significantly higher than those in other groups (P<0.01), indicating that the composition of the present invention can achieve the effect of promoting lactation by upregulating the levels of lactation-related hormones.

[0065] 6. Experimental Conclusion: The lactation-promoting composition for dairy cows prepared in this invention can significantly increase milk yield, improve milk quality, regulate the secretion of lactation-related hormones, and protect mammary gland health. Its effects are significantly better than existing soy isoflavone products, and it has extremely high application value in animal husbandry.

[0066] Experiment Example 4 Experiment on the effect of the composition on the lactation performance of sows: This experimental example verifies the effect of the oral lactation-promoting liquid for sows prepared in Example 3 on the lactation performance of sows.

[0067] 1. Experimental animals and grouping: Forty large-scale crossbred sows from a large-scale pig farm in Hunan Province were selected. They were of 2nd to 4th parity, with similar expected farrowing dates, and were healthy and disease-free. The sows were randomly divided into 4 groups of 10 sows each. The grouping was the same as in Experiment 3. The administration method was as follows: oral liquid was administered by gavage starting from the day of farrowing, 20 mL / sow / day, for 21 consecutive days (during lactation). The basic diet and feeding management conditions of the sows in each group were completely identical.

[0068] 2. Detection indicators: (1) Sow milk production: The difference in piglet weight was used to weigh the difference in litter weight before and after lactation at a fixed time every day to calculate the average daily milk production of the sow; (2) Piglet growth performance: The birth litter weight, weaning litter weight at 21 days of age, survival rate, and average daily weight gain of piglets were recorded; (3) Sow serum hormone indicators: Blood was collected from the anterior vena cava of sows on the 7th, 14th and 21st days after farrowing, serum was separated, and prolactin content was detected by ELISA; (4) Sow postpartum recovery: The interval between postpartum estrus and the incidence of constipation during lactation were recorded.

[0069] 3. Experimental results: (1) Effect on sow milk production: During the experiment, the average daily milk production of sows in Example 3 group was 8.6±0.5kg, while that in the blank control group was 6.2±0.4kg, an increase of 38.71%, which was significantly higher than other groups (P<0.01); the average daily milk production of Comparative Example 2 group was 7.1±0.4kg, an increase of 14.52% compared with the blank control group, which was significantly lower than that in Example 3 group (P<0.05). (2) Effect on piglet growth performance: The growth performance results of piglets in each group are shown in Table 3.

[0070] Table 3 Comparison of growth performance of piglets in each group As shown in Table 3, the litter weight of piglets weaned at 21 days of age in the Example 3 group was 42.12% higher than that in the blank control group, the survival rate of piglets was increased to 98.5%, and the average daily weight gain was increased by 52.20%, which was significantly better than other groups (P<0.01). (3) Effect on postpartum recovery of sows: The estrus interval of sows in the Example 3 group was 5.2±0.3 days, which was significantly shorter than that of the blank control group (7.8±0.5 days) (P<0.05); the incidence of constipation during lactation was 5%, which was significantly lower than that of the blank control group (30%) (P<0.05), indicating that the composition of the present invention can significantly promote the postpartum recovery of sows.

[0071] 4. Experimental conclusions: The oral liquid for promoting lactation in sows prepared in this invention can significantly increase the milk production of sows, promote the growth and development of piglets, improve the survival rate, and accelerate the postpartum recovery of sows. The effect is significantly better than that of existing similar products, and it is suitable for promotion and application in large-scale pig farms.

[0072] Experiment Example 4 This study investigates the molecular mechanism by which the composition promotes lactation using a rat lactation model.

[0073] 1. Experimental animals and grouping: 30 SPF-grade SD female pregnant rats, 18 days pregnant and weighing 280-320g, were selected and provided by Hunan Slack Jingda Experimental Animal Co., Ltd. The rats were randomly divided into 3 groups of 10 rats each: (1) Blank control group: 0.5% sodium carboxymethyl cellulose solution was administered by gavage after birth; (2) Low-dose group: 100mg / kg body weight / day of lime flavonoid extract was administered by gavage after birth; (3) High-dose group: 300mg / kg body weight / day of lime flavonoid extract was administered by gavage after birth. The rats in each group were gavaged by gavage from the day of birth for 7 consecutive days. During this period, they were allowed free access to food and water. The number of pups per litter was adjusted to 8 to ensure consistent lactation load.

[0074] 2. Detection indicators: (1) Rat milk production: The average daily milk production of rats was calculated using the difference in pups' weight. (2) Observation of mammary gland tissue morphology: On the 7th day of the experiment, rats were euthanized, and the 4th pair of mammary gland tissues were taken, fixed with 4% paraformaldehyde, embedded in paraffin, sectioned, stained with HE, and observed under an optical microscope to observe the development of mammary gland alveoli. (3) Real-time quantitative PCR (qRT-PCR): Mammary gland tissue was taken, total RNA was extracted, reverse transcribed into cDNA, and qRT-PCR was used to detect the mRNA expression levels of prolactin receptor (PRLR), estrogen receptor α (ERα), estrogen receptor β (ERβ), and β-casein genes in mammary gland tissue. GAPDH was used as an internal reference gene. (4) Western blot detection: Total protein was extracted from mammary gland tissue, and Western blot was used to detect the protein expression levels of PRLR, ERα, and β-casein.

[0075] 3. Experimental results: (1) Milk production in rats: After 7 days of continuous gavage, the average daily milk production of rats in the low-dose group was 26.5±1.8g and that in the high-dose group was 32.8±2.1g, which were 25.00% and 54.72% higher than that in the blank control group (21.2±1.5g), respectively. The difference between the groups was extremely significant (P<0.01), showing obvious dose dependence. (2) Mammary gland morphology: The number of mammary alveoli in rats in the blank control group was small, the alveolar cavities were small, and the mammary gland interstitial fat tissue was abundant. The number of mammary alveoli in the low-dose group and the high-dose group was significantly increased, the alveolar cavities were significantly enlarged, the alveolar epithelial cells were neatly arranged, the secretory function was vigorous, and the degree of mammary gland development was significantly better than that in the blank control group. The effect of the high-dose group was more obvious. (3) Gene expression results: Compared with the blank control group, the mRNA expression levels of PRLR, ERα, ERβ and β-casein in the mammary tissue of rats in both the low-dose and high-dose groups were significantly upregulated (P<0.05), and the upregulation in the high-dose group was significantly higher than that in the low-dose group (P<0.05), indicating that the flavonoid extract of lime can upregulate the expression of lactation-related genes at the transcriptional level. (4) Protein expression results: Western blot results showed that the protein expression levels of PRLR, ERα and β-casein in the mammary tissue of rats in both the low-dose and high-dose groups were significantly higher than those in the blank control group (P<0.05), which was consistent with the gene expression results, further verifying the target of the flavonoid extract of lime.

[0076] 4. Experimental conclusions: The flavonoid extract of lime of the present invention can promote the development of mammary alveoli by upregulating the gene and protein expression levels of lactation-related receptors and functional proteins in mammary tissue, thereby achieving the effect of promoting lactation at the molecular level, and clarifying the core mechanism of action of the composition of the present invention.

[0077] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific structures and characteristics of the solutions is not described in detail here. It will be apparent to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the present invention is defined by the appended claims rather than the foregoing description. Therefore, it is intended that all variations falling within the meaning and scope of equivalents of the claims be included within the present invention. No markings in the claims should be construed as limiting the scope of the claims.

Claims

1. A lactation-promoting composition containing flavonoid extract of lime, characterized in that, By weight, it includes the following raw material components: 10-50 parts of lime flavonoid extract, 5-30 parts of synergistic active ingredients, and 20-85 parts of excipients; wherein the lime flavonoid extract contains ≥80% total flavonoids based on hesperidin, of which hesperidin ≥40%, neohesperidin ≥15%, naringin ≥10%, and hesperidin ≥5%.

2. The lactation-promoting composition according to claim 1, characterized in that, The preparation method of the lime flavonoid extract includes the following steps: (1) Raw material pretreatment: Take dried and ripe sour oranges, crush them through a 40-mesh sieve to obtain sour orange raw material powder; (2) Degreasing pretreatment: The lime raw material powder was degreased twice by reflux with petroleum ether at 60°C for 2 hours each time. After filtration, the petroleum ether was evaporated from the filter residue to obtain degreased lime powder. (3) Ultra-high pressure assisted ethanol extraction: Add 70% volume fraction of ethanol aqueous solution to defatted lime powder, with a material-liquid ratio of 1:

15. After mixing evenly, transfer to an ultra-high pressure extraction device, set the ultra-high pressure to 300 MPa and the extraction temperature to 50℃, extract 3 times, 40 min each time. After extraction, collect and combine the extracts, and recover ethanol under reduced pressure to obtain flavonoid concentrate. (4) Macroporous resin purification: Dilute the flavonoid concentrate to a total flavonoid concentration of 2 mg / mL, load the sample onto an AB-8 macroporous resin column at a flow rate of 2 BV / h and a loading volume of 4 BV; wash with 5 BV of deionized water to remove impurities, and then wash with 6 BV of 70% ethanol aqueous solution and collect the ethanol eluent. (5) Concentration and drying: The ethanol eluent was recovered under reduced pressure and then freeze-dried under vacuum to obtain the flavonoid extract of orange.

3. The lactation-promoting composition according to claim 1, characterized in that, The synergistic active ingredients are one or more of the following: Wangbuliuxing flavonoids, Angelica sinensis polysaccharides, Codonopsis pilosula polysaccharides, Astragaloside A, Astragalus polysaccharides, vitamin E, folic acid, β-carotene, and probiotic freeze-dried powder.

4. The lactation-promoting composition according to claim 1, characterized in that, The composition is available in the form of a livestock feed premix, soluble powder, oral liquid, or granules.

5. The lactation-promoting composition according to claim 4, characterized in that, The excipients for the animal husbandry formulation are one or more of the following: corn starch, wheat bran, skim milk powder, limestone powder, maltodextrin, Tween-80, potassium sorbate, white sugar, and sodium carboxymethyl cellulose.

6. A method for preparing the lactation-promoting composition containing flavonoid extract of orange as described in claim 1, characterized in that, When the composition is in solid dosage form, the following steps are included: (1) Raw material pretreatment: Pulverize the lime flavonoid extract, synergistic active ingredients and excipients separately, pass them through an 80-100 mesh sieve, and weigh them according to the formula weight parts for later use; (2) Mixing: Place the weighed raw materials in a three-dimensional motion mixer and mix for 20-40 minutes until the mixing uniformity RSD ≤ 5% to obtain mixed powder; (3) Forming and Packaging: According to the dosage form requirements, the mixed powder is directly packaged, or it is formed after granulation, drying, sizing, filling / tableting, and then packaged after passing quality inspection to obtain the finished product.

7. A method for preparing the lactation-promoting composition containing flavonoid extract of orange as described in claim 1, characterized in that, When the composition is in the form of an oral liquid dosage form Includes the following steps: (1) Raw material pretreatment: Weigh each raw material according to the formula weight parts and set aside; (2) Solution preparation: Take 80% of the prescription amount of deionized water, add solubilizer, stir evenly, add lime flavonoid extract and synergistic active ingredients, stir until completely dissolved; then add the remaining excipients, stir until completely dissolved, add deionized water to the total amount to obtain the drug solution; (3) Filtration, sterilization and packaging: The liquid medicine is filtered through a 0.45μm microporous membrane, filled, capped and sterilized. After passing quality inspection, it is packaged to obtain the finished product.

8. The use of the lactation-promoting composition containing flavonoid extract of orange as described in claim 1 in the preparation of lactation-promoting products.

9. The application according to claim 8, characterized in that, The lactation-promoting products are lactation-promoting feed additives and veterinary drug preparations for livestock breeding, and the livestock include dairy cows, sows, meat sheep, and rabbits.

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