CN118160831A - Rumen-bypass melatonin product and preparation method and application thereof - Google Patents

Rumen-bypass melatonin product and preparation method and application thereof Download PDF

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Publication number
CN118160831A
CN118160831A CN202410398999.3A CN202410398999A CN118160831A CN 118160831 A CN118160831 A CN 118160831A CN 202410398999 A CN202410398999 A CN 202410398999A CN 118160831 A CN118160831 A CN 118160831A
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melatonin
rumen
wall material
bypass
product
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马慧
麻柱
王桂英
吕晓莲
阚远征
白沥冰
初晓宁
杨宇泽
李凯扬
付瑶
王俊
陈少康
罗维真
潘学峰
张红涛
刘春海
陈凤林
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Beijing Sanyuan Seed Industry Technology Co ltd
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Beijing Sanyuan Seed Industry Technology Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/116—Heterocyclic compounds
    • A23K20/132—Heterocyclic compounds containing only one nitrogen as hetero atom
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/02—Breeding vertebrates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/158—Fatty acids; Fats; Products containing oils or fats
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/184—Hormones
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A23K40/10—Shaping or working-up of animal feeding-stuffs by agglomeration; by granulation, e.g. making powders
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Zoology (AREA)
  • Food Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Husbandry (AREA)
  • Environmental Sciences (AREA)
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  • Animal Behavior & Ethology (AREA)
  • Health & Medical Sciences (AREA)
  • Endocrinology (AREA)
  • Fodder In General (AREA)

Abstract

The invention provides a rumen-bypass melatonin product, and a preparation method and application thereof, and belongs to the technical field of livestock breeding, wherein the rumen-bypass melatonin product comprises a core material, a first wall material and a second wall material, and the first wall material and the second wall material are sequentially coated outside the core material; the core material is melatonin, and the first wall material and the second wall material both comprise stearic acid and hypromellose; the preparation method comprises the steps of firstly preparing melatonin fluidization pellets, coating a first wall material, and coating a second wall material; the prepared rumen-bypass melatonin product is used for feeding cows, so that the number of somatic cells of cow milk is reduced, the protein content of the cow milk is increased, and the cow milk yield is increased. The rumen-bypass melatonin product prepared by the invention has high rumen bypass rate, high small intestine release rate and good application effect of melatonin.

Description

Rumen-bypass melatonin product and preparation method and application thereof
Technical Field
The invention relates to melatonin and application thereof, in particular to a rumen-bypass melatonin product and application thereof, and belongs to the technical field of livestock breeding.
Background
With the increasing demand of people for dairy products, the dairy industry is gradually growing, and large-scale dairy farms are increasing. However, as the size of the cow farm is enlarged, the cow breeding problem is gradually highlighted, wherein the most harmful problem is the problem of high somatic cell count of the cow. The high and low of the number of the somatic cells of the dairy cows closely relate to the health condition of the breast and the milk yield, and huge economic loss is brought to dairy cow breeding enterprises each year due to the problem of the number of the somatic cells of the dairy cows. Milk components in milk in China, especially milk with high milk protein content, have higher nutritive value than milk with the milk protein content in developed countries.
Melatonin is an amine neuroendocrine hormone secreted by the pine cone body and derived from tryptophan, also called melanine, melanostatin, pineal pancreas extract and the like, is an endogenous substance which is self-produced by animals, plants and microorganisms, is insoluble in water, can be dissolved in organic solvents such as acetone and the like, can act on a plurality of organs, tissues and cells of the whole body of the animal body, and has various physiological and pharmacological effects of regulating biological rhythm, improving sleep, regulating reproductive activity, resisting oxidation, resisting tumors, scavenging free radicals in the body and the like.
The main source of melatonin is chemical synthesis or natural secretion of organism, but the self secretion of organism is little, can only satisfy the needs of animal organism itself. Because melatonin synthesis and extraction processes are complicated, the cost is high, the application in livestock production is limited to test animals, and the batch production of livestock products is limited. Meanwhile, the existing melatonin used in livestock production is mostly given to animals in an injection mode, so that the operation is inconvenient and the cost is high.
Because of the special digestive system, nutrients are degraded in large amounts in the rumen of ruminants, and therefore, the direct addition of melatonin to ruminants cannot achieve the desired purpose. In order for melatonin to exert its intended effects and effects in ruminant production practice, rumen-protected melatonin is required. The rumen bypass technology protects some nutrient substances which are easy to be destroyed by rumen microorganisms by using a physical or chemical method from being decomposed by the rumen microorganisms, well passes through the rumen, reaches abomasum and intestinal tracts and is released again, and plays a role in the small intestine, so that the requirement of organisms on the nutrient substances is met.
The patent CN 110447778A-rumen-bypass melatonin is applied to improving the structure of rumen microorganism flora of ruminants and improving milk quality, and the rumen-bypass melatonin can reduce the number of somatic cells in milk of ruminants, can improve the content of milk protein, can improve the milk fat percentage, can reduce the lactose content and the like. The rumen-bypass melatonin consists of a coating and a cladding, wherein the coating is rumen-bypass fat powder, and the active ingredient of the cladding is melatonin; the coating core is particles prepared from active ingredients melatonin and auxiliary materials; the auxiliary materials comprise a first auxiliary material and a second auxiliary material. The first auxiliary material is calcium stearate or silicon dioxide, and the second auxiliary material is composed of starch, dextrin and sodium carboxymethyl cellulose. The mass ratio of the coating core to the coating is 45:55-50:50, and through a fistula cow experiment, the rumen bypass melatonin has the rumen bypass rate of 83-85%. The rumen-bypass melatonin product has various auxiliary materials, low rumen-bypass rate, high melatonin loss and unknown release rate in small intestine.
Disclosure of Invention
Aiming at the problems, the invention provides a rumen-bypass melatonin product, a preparation method and application thereof, and the rumen-bypass melatonin product can improve the rumen-bypass rate of rumen-bypass melatonin and the small intestine release rate of rumen-bypass melatonin, so that the application effect of melatonin is improved.
In order to achieve the above purpose, the technical scheme of the invention is as follows: the rumen-bypass melatonin product comprises a core material, a first wall material and a second wall material, wherein the first wall material and the second wall material are sequentially coated outside the core material; the core material is melatonin, and the first wall material and the second wall material both comprise stearic acid and hypromellose.
Further, the mass ratio of melatonin to the first wall material to the second wall material in the rumen-bypass melatonin product is 0.9-1.1:0.9-1.1:0.9-1.1.
Further, the mass ratio of the stearic acid to the hypromellose in the first wall material is 1:2-3, and the mass ratio of the stearic acid to the hypromellose in the second wall material is 2-3:1.
The preparation method of the rumen-bypass melatonin product comprises the following steps:
s1, taking melatonin, adding a proper amount of water, uniformly mixing, preparing into strip-shaped particles in a granulator, transferring into a shot blasting machine, rotating, boiling and blasting, and drying to obtain melatonin pellets; transferring melatonin pellets into a fluidized bed for fluidization treatment to obtain fluidized pellets;
s2, heating the first wall material to 120 ℃ for melting, cooling to 80-100 ℃, and coating the first wall material on the surface of the fluidized pellets to obtain rumen melatonin intermediates;
and S3, heating the second wall material to 120 ℃ for melting, cooling to 80-100 ℃, and coating the second wall material on the surface of the rumen-bypass melatonin intermediate to obtain the rumen-bypass melatonin product.
Further, the moisture content of the melatonin pellets after drying in the step S1 is less than or equal to 10wt%.
Further, in the step S1, the atomization pressure of the fluidized bed is 3.5kg/cm 2, the rotating speed is 90-95r/min, the air inlet temperature is 35-45 ℃, and the air outlet temperature is 25-45 ℃.
The rumen-bypass melatonin product is applied to feeding cows, so that the number of somatic cells of cow milk is reduced, the protein content of the cow milk is increased, and the cow milk yield is increased.
Further, the variety of the dairy cows is Holstein dairy cows.
Further, each cow is fed 400-450mg rumen-bypass melatonin product daily.
Furthermore, after the rumen-bypass melatonin product is fed to the dairy cows, the protein content of the dairy cows is improved by 30-40%, the number of somatic cells is reduced by 60-70%, and the yield of the dairy cows is improved by 10-15% compared with the dairy cows without the rumen-bypass melatonin product.
The rumen-bypass melatonin product and the preparation method and application thereof have the beneficial effects that:
The rumen-bypass melatonin product prepared by the invention has high rumen bypass rate, high small intestine release rate and good application effect of melatonin. The rumen-bypass melatonin product prepared by the invention is used for feeding cows, so that the number of somatic cells of cow milk can be reduced, the protein content of the cow milk can be increased, and the cow milk yield can be increased.
The wall material used by the rumen by-pass melatonin is stearic acid and hypromellose, wherein the stearic acid is also called octadecanoic acid, is produced by oil hydrolysis, and is pure white crystals, insoluble in water and soluble in organic solvents; belongs to long-chain saturated fatty acid and has stable chemical property. The hydroxypropyl methylcellulose is prepared by specially etherifying high-purity cotton cellulose under alkaline conditions, is white powder, can be dissolved in water, and has the characteristics of certain film forming property, enzyme resistance and the like. The invention uses stearic acid or hypromellose as wall material to coat melatonin, which has extremely high degradation rate in rumen, and has great melatonin loss and great influence on intestinal absorption of melatonin.
The rumen-bypass melatonin is coated by two layers of wall materials, so that the rumen-bypass rate of the melatonin in the rumen is improved, and the release rate of the melatonin in the small intestine is also improved. The first wall material adopts the combination of relatively high-content hypromellose and relatively low-content stearic acid, the second wall material of the outermost layer adopts the combination of relatively high-content stearic acid and relatively low-content hypromellose, as the rumen of the cattle is obviously different from the small intestine environment, the pH value is greatly different (the rumen environment is close to neutral and the small intestine environment is strongly acidic), the second wall material of the outermost layer has high stearic acid content and stable chemical property, has a certain coating effect in combination with the hypromellose and has certain tackiness, and the interaction of the two promotes the coating effect of the second wall material, reduces the degradation rate in the rumen of the cattle, and simultaneously overcomes the defect that the effect of singly taking stearic acid as the wall material coating is poor; the second wall material is degraded to a certain extent after passing through the rumen of the cattle, and after melatonin reaches the small intestine of the cattle, the wall material is required to be rapidly degraded to explain the change of environments such as pH value and the like, the melatonin placed in the wall material is required to be absorbed by the small intestine, the first wall material is positioned on the inner side of the second wall material, the second wall material is degraded firstly, the first wall material is degraded secondly, the content of hydroxypropyl methylcellulose in the first wall material is higher, and the combination of the hydroxypropyl methylcellulose with stearic acid with high content is easier to degrade in a strong acid environment, so that the melatonin is rapidly released, and the effect of improving the release rate of the melatonin in the small intestine is achieved.
The wall material coated with melatonin is easy to obtain and low in cost; the rumen-bypass melatonin product has high melatonin content and low wall material content, so that the coating cost is reduced; in addition, the coating material has a lower content, but the coating effect is much better than that of a single coating material.
The rumen-bypass melatonin prepared by the method has the advantages that the rumen-bypass melatonin is furthest protected by the wall materials, the melatonin loss is extremely small, the melatonin can be rapidly and completely released in the bovine intestinal environment, and the two wall materials are completely degraded and completely released, so that the melatonin can be fully absorbed by the intestinal tract, and the dairy quality of the dairy cows can be improved.
In addition, the preparation method is simple, has high production efficiency and is suitable for large-scale production.
Detailed Description
The following description of the technical solution in the embodiments of the present invention is clear and complete. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways other than those described herein, and persons skilled in the art will readily appreciate that the present invention is not limited to the specific embodiments disclosed below.
The rumen-bypass melatonin product comprises a core material, a first wall material and a second wall material, wherein the first wall material and the second wall material are sequentially coated outside the core material; the core material is melatonin, and the first wall material and the second wall material both comprise stearic acid and hypromellose. Preferably, the mass ratio of melatonin to the first wall material to the second wall material in the rumen-bypass melatonin product is 0.9-1.1:0.9-1.1:0.9-1.1. Further preferably, the mass ratio of the stearic acid to the hypromellose in the first wall material is 1:2-3, and the mass ratio of the stearic acid to the hypromellose in the second wall material is 2-3:1.
The preparation method of the rumen-bypass melatonin product comprises the following steps:
S1, taking melatonin, adding a proper amount of water, uniformly mixing, preparing into strip-shaped particles in a granulator, transferring into a shot blasting machine, rotating, boiling and blasting, and drying to obtain melatonin pellets; transferring melatonin pellets into a fluidized bed for fluidization treatment to obtain fluidized pellets; preferably, the moisture content of the dried melatonin micropills is less than or equal to 10wt%; the atomization pressure of the fluidized bed is 3.5kg/cm 2, the rotating speed is 90-95r/min, the air inlet temperature is 35-45 ℃, and the air outlet temperature is 25-45 ℃ during fluidization treatment;
s2, heating the first wall material to 120 ℃ for melting, cooling to 80-100 ℃, and coating the first wall material on the surface of the fluidized pellets to obtain rumen melatonin intermediates;
and S3, heating the second wall material to 120 ℃ for melting, cooling to 80-100 ℃, and coating the second wall material on the surface of the rumen-bypass melatonin intermediate to obtain the rumen-bypass melatonin product.
The rumen-bypass melatonin product is applied to feeding cows, so that the number of somatic cells of cow milk is reduced, the protein content of the cow milk is increased, and the cow milk yield is increased. Preferably, the variety of the dairy cows is Holstein dairy cows; further preferably, 400-450mg by-pass rumen melatonin product is fed per cow per day; further preferably, after the rumen-bypass melatonin product is fed to cows, the content of milk protein of cows is increased by 30-40%, the number of somatic cells is reduced by 60-70%, and the yield of cows is increased by 10-15% compared with cows not fed with the rumen-bypass melatonin product.
Example 1
Mixing 35kg melatonin and 7kg water uniformly, preparing strip-shaped particles in a granulator, then rotating boiling and shot blasting in a shot blasting machine, and drying for 80 minutes at the temperature of 50 ℃ until the water content is less than or equal to 10wt% to obtain dried melatonin pellets. Pouring the dried melatonin pellets into a storage tank of a fluidized bed, adjusting the atomization pressure to 3.5kg/cm < 2 >, and obtaining fluidized pellets, wherein the rotating speed is 90 turns/min, the air inlet temperature is 35 ℃, and the air outlet temperature is 25 ℃.
Taking 11kg of stearic acid and 22kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 80 ℃, and coating the mixture on the surface of the fluidized pellets to obtain the rumen melatonin intermediate.
Taking 23.3kg of stearic acid and 11.7kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 80 ℃, and coating the mixture on the surface of the fluidized pellets to obtain rumen-protected melatonin, wherein the mass concentration of the melatonin is about 30%.
Example 2
Mixing 35kg melatonin and 7kg water uniformly, preparing strip-shaped particles in a granulator, then rotating boiling and shot blasting in a shot blasting machine, and drying for 80 minutes at the temperature of 50 ℃ until the water content is less than or equal to 10wt% to obtain dried melatonin pellets. Pouring the dried melatonin pellets into a storage tank of a fluidized bed, adjusting the atomization pressure to 3.5kg/cm < 2 >, and obtaining fluidized pellets, wherein the rotating speed is 90 turns/min, the air inlet temperature is 35 ℃, and the air outlet temperature is 25 ℃.
Taking 11.7kg of stearic acid and 23.3kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 100 ℃, and coating the mixture on the surface of the fluidized pellets to obtain the rumen melatonin intermediate.
Taking 22kg of stearic acid and 11kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 100 ℃, and coating the mixture on the surface of the fluidized pellets, wherein the rumen-protected melatonin is about 30% in mass concentration.
Example 3
Taking 38kg of melatonin and 8kg of water, uniformly mixing, preparing into strip-shaped particles in a granulator, then rotating in a shot blasting machine to boil and throw balls, and drying for 80 minutes at the temperature of 50 ℃ until the water content is less than or equal to 10wt%, thereby obtaining the dried melatonin pellets. Pouring the dried melatonin pellets into a storage tank of a fluidized bed, adjusting the atomization pressure to 3.5kg/cm < 2 >, and obtaining fluidized pellets, wherein the rotating speed is 90 turns/min, the air inlet temperature is 35 ℃, and the air outlet temperature is 25 ℃.
Taking 12kg of stearic acid and 24kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 110 ℃, and coating the mixture on the surface of the fluidized pellets to obtain the rumen melatonin intermediate.
Taking 23.3kg of stearic acid and 11.7kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 110 ℃, and coating the mixture on the surface of the fluidized pellets to obtain rumen-protected melatonin, wherein the mass concentration of the melatonin is about 34%.
Example 4
Taking 38kg of melatonin and 8kg of water, uniformly mixing, preparing into strip-shaped particles in a granulator, then rotating in a shot blasting machine to boil and throw balls, and drying for 80 minutes at the temperature of 50 ℃ until the water content is less than or equal to 10wt%, thereby obtaining the dried melatonin pellets. Pouring the dried melatonin pellets into a storage tank of a fluidized bed, adjusting the atomization pressure to 3.5kg/cm < 2 >, and obtaining fluidized pellets, wherein the rotating speed is 90 turns/min, the air inlet temperature is 35 ℃, and the air outlet temperature is 25 ℃.
Taking 13kg of stearic acid and 26kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 90 ℃, and coating the mixture on the surface of the fluidized pellets to obtain the rumen melatonin intermediate.
Heating 26kg of stearic acid and 13kg of hypromellose to 120 ℃ for melt mixing, cooling to 90 ℃, and coating the mixture on the surface of the fluidized pellets to obtain rumen-protected melatonin, wherein the mass concentration of the melatonin is about 30%.
Example 5
Mixing 35kg melatonin and 7kg water uniformly, preparing strip-shaped particles in a granulator, then rotating boiling and shot blasting in a shot blasting machine, and drying for 80 minutes at the temperature of 50 ℃ until the water content is less than or equal to 10wt% to obtain dried melatonin pellets. Pouring the dried melatonin pellets into a storage tank of a fluidized bed, adjusting the atomization pressure to 3.5kg/cm < 2 >, and obtaining fluidized pellets, wherein the rotating speed is 90 turns/min, the air inlet temperature is 35 ℃, and the air outlet temperature is 25 ℃.
Taking 8kg of stearic acid and 24kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 80 ℃, and coating the mixture on the surface of the fluidized pellets to obtain the rumen melatonin intermediate.
Taking 24kg of stearic acid and 8kg of hypromellose, heating to 120 ℃ for melt mixing, cooling to 80 ℃, and coating the mixture on the surface of the fluidized pellets, wherein the rumen-protected melatonin is about 34% in mass concentration.
Example 6
Mixing 40kg melatonin and 8kg water uniformly, preparing strip-shaped particles in a granulator, then rotating boiling and shot blasting in a shot blasting machine, and drying for 80 minutes at the temperature of 50 ℃ until the water content is less than or equal to 10wt% to obtain dried melatonin pellets. Pouring the dried melatonin pellets into a storage tank of a fluidized bed, adjusting the atomization pressure to 3.5kg/cm < 2 >, and obtaining fluidized pellets, wherein the rotating speed is 90 turns/min, the air inlet temperature is 35 ℃, and the air outlet temperature is 25 ℃.
Heating 10kg stearic acid and 30kg hypromellose to 120deg.C, mixing, cooling to 110deg.C, and coating on the surface of fluidized pellet to obtain rumen melatonin intermediate.
Heating 30kg stearic acid and 10kg hypromellose to 120deg.C, mixing, cooling to 110deg.C, and coating on the surface of fluidized pellet to obtain rumen melatonin with mass concentration of about 30%.
Test example 1 effect verification
In order to verify the performance of the rumen protected melatonin prepared by the invention, the stability test and evaluation are performed by simulating the digestive tract of ruminant animals by an in vitro method (InVitro).
1. Materials and methods
The rumen environment and the duodenum environment of ruminants were simulated respectively using a buffer with a pH of 6.6 and a buffer with a pH of 2.4. The formulations of the buffers at different pH values are shown in Table 1.
TABLE 1 buffer formulations (unit: g) for different pH values
Reagent(s) PH value of 6.6 PH value is 2.4
Citric acid 5.7225 19.698
Disodium hydrogen phosphate 52.089 4.4392
The materials listed in Table 1 were dissolved in a small amount of distilled water to a volume of 1000 ml.
2. Test sample
The rumen-protected melatonin prepared in examples 1-6 was selected for a total of 6 samples. Each sample was prepared in 3 production batches of 400g each.
3. Stability test of rumen melatonin in different pH buffers
Respectively accurately weighing 1.00g of each of the 6 samples, placing at the bottom of a 50ml test tube with a plug, adding 20ml buffer solution, and tightly covering the test tube plug; digesting for 2, 4, 8, 12, 24, 48 and 72 hours in a constant temperature water bath shaker at 39 ℃; taking out, washing, filtering, fixing the volume of the filtrate, and measuring the content of melatonin in the filtrate, thereby calculating the rumen bypass rate and the small intestine release rate of the melatonin. Three replicates were set for each coated rumen melatonin at each time point.
4. Melatonin detection method
Melatonin content was determined with reference to GB/T5009.170. The calculation formula is as follows:
rumen bypass rate (W1) = (A1-A2)/a1×100% of the product
Wherein: a1, melatonin content in the product;
A2-melatonin content of the product in the buffer filtrate at pH 6.6.
Product small intestine release rate (W2) =a3/a1×100%
Wherein: a1, melatonin content in the product;
a3-melatonin content of the product in the filtrate of the buffer with pH value of 2.4.
The statistical method comprises the following steps: data analysis was performed using SPSS 19.0.
5. Analysis of results
(1) Rumen bypass rate (pH 6.6) at various time points for each product is shown in table 2.
TABLE 2 rumen bypass Rate (%)
Time/sample 2h 4h 8h 12h 24h 48h 72h
Example 1 99.88 99.21 97.89 96.78 95.47 93.26 91.57
Example 2 99.92 99.31 98.01 96.89 95.34 93.19 91.73
Example 3 99.34 99.02 97.45 96.23 94.79 93.01 90.98
Example 4 99.36 99.11 97.39 96.43 94.58 93.07 91.06
Example 5 99.67 99.29 97.67 96.35 95.12 93.38 91.27
Example 6 99.33 99.01 97.82 96.44 95.16 93.09 90.11
The results shown in Table 2 were obtained by simulating the rumen environment of ruminants with a pH 6.6 buffer and incubating each test sample in a 39℃constant temperature water bath. From table 2, it can be seen that the rumen bypass melatonin prepared by the embodiment of the invention has high rumen bypass rate, the rumen bypass rate after 72 hours is still more than ninety percent, and the rumen bypass melatonin has stable properties.
(2) The small intestine release rate (ph=2.4) at various time points for each product is shown in table 3.
TABLE 3 Small intestine Release Rate (%)
The results shown in Table 3 were obtained by simulating the duodenal environment of ruminants with a pH 2.4 buffer and incubating each test sample in a 39℃constant temperature water bath. From table 3, it can be seen that the rumen-bypass melatonin prepared by the embodiment of the invention has high release rate of small intestine, and each sample basically realizes complete release at 4 hours.
Test example 2 methodological verification
The effect was verified by the following method for measuring the rumen bypass rate and the small intestine release rate in the effect verification of test example 1, and only the rumen bypass rate and the small intestine release rate at 2 hours, 4 hours, 8 hours and 12 hours were examined.
1) Selection of wall material
Stearic acid is a wall material commonly used in the existing rumen bypass coating technology, is produced by grease hydrolysis, is insoluble in water and organic solvent, belongs to long-chain saturated fatty acid, and has stable chemical property. In the prior art, the stearic acid is used for coating the lysine, the passing rate is higher after the stearic acid is cultured in the rumen, the stearic acid is used as the wall material of the rumen-bypass melatonin in the experiment,
The condensing method (namely the preparation method of the embodiment of the invention) commonly used in the field is adopted for coating the melatonin, the mass ratio of the melatonin to the stearic acid is set to be 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 and 1:5, and the rumen bypass rate results show (table 4) that the rumen bypass rate tends to be increased along with the increase of the stearic acid ratio when the mass ratio of the melatonin to the stearic acid is 5:1 to 1:2, and the high wall material content can slow down the exposure of the core material, so that the rumen bypass rate is improved; the rumen bypass rate is not greatly different from that of 1:2 when the mass ratio of melatonin to stearic acid is 1:3, and the rumen bypass rate is not increased when the mass ratio reaches 1:4 and 1:5, which indicates that the coating effect of the wall material can be improved by increasing the content of the wall material without taste, and the coating effect is related to the property of the wall material, such as degradation difficulty, pH sensitivity and the like; however, when the ratio of melatonin to stearic acid is 1:2 or 1:3 with better rumen bypass effect, the rumen bypass rate is still less than 60% at 2h, and the rumen bypass rate is reduced to below 50% at 8h, which indicates that the effect of stearic acid serving as a wall material on the rumen bypass rate of melatonin is poor and still needs to be optimized.
TABLE 4 rumen bypass Rate (%)
Proportion of 2h 4h 8h 12h
5:1 21.33 16.51 9.73 5.38
4:1 22.46 19.72 15.35 9.47
3:1 25.72 20.31 18.72 15.31
2:1 30.44 25.36 21.33 18.42
1:1 41.79 39.26 32.16 24.78
1:2 57.89 51.35 47.92 43.26
1:3 58.02 51.77 48.56 42.17
1:4 57.99 52.06 48.13 42.69
1:5 58.10 51.33 47.96 42.78
The problem of low rumen bypass rate also occurs when the methionine is coated by stearic acid by the existing researchers, the rumen bypass rate is obviously increased after a certain amount of bentonite is added, the bentonite is non-metal mineral products taking montmorillonite as a main mineral component, the bentonite has certain viscosity, and the viscosity of the bentonite is considered to improve the coating effect of the stearic acid.
2) Screening of adhesives
Based on the test results, bentonite, starch and hypromellose with certain adhesiveness are selected and respectively added into the wall material, and the adhesive with better screening effect is selected. And adding a certain amount of the three binders on the basis of the mass ratio of melatonin to stearic acid being 1:2, and performing screening test according to the mass ratio of melatonin to stearic acid to binder being 1:2 (0.5 or 1 or 1.5). The test results (table 5 below) show that the rumen bypass rate is improved to a certain extent after bentonite and starch are added, but the improvement range is not large, and the melatonin degradation is still serious; the rumen bypass rate is obviously improved after the hypromellose is added, the rumen bypass rate is firstly improved and then reduced along with the increase of the adding amount of the hypromellose, and when the melatonin is prepared from the following components: 2: the rumen bypass rate is relatively highest in the step 1, and the rumen bypass rate is still higher after 12 hours, which indicates that the property of the wall material is stable; in addition, under the condition of the optimal proportion, the wall material occupies a relatively high proportion, which is probably one reason for longer property stability time, and the content of the wall material can be further reduced, so that the content of melatonin is improved; the hydroxypropyl methylcellulose can be dissolved in water, has a wider pH value stability range (the pH value is stable when the pH value is 4-10), has certain cohesiveness, and is hypothesized that the cooperation of the cohesiveness and stearic acid improves the rumen bypass rate of stearic acid serving as a wall material, and the pH value stability of the hydroxypropyl methylcellulose further improves the rumen bypass rate, so that the cooperation of stearic acid and the hydroxypropyl methylcellulose is selected as the wall material.
TABLE 5 rumen bypass Rate (%)
Wherein, the proportion of the addition amount in the column is melatonin: stearic acid: and (3) an adhesive.
3) Wall material addition amount screening
Preparing a wall material according to the mass ratio of stearic acid to hypromellose of 2:1, setting the mass ratio of melatonin to the wall material to be 1:0.5, 1:1, 1:1.5, 1:2 and 1:3, preparing rumen-bypass melatonin according to the method of the embodiment, verifying the rumen-bypass rate, and as shown in the following table 6, the result shows that the rumen-bypass melatonin has higher rumen-bypass rate when the mass ratio of melatonin to the wall material reaches 1:1-1:1.5, the increase of the rumen-bypass rate of the wall material is not obvious, and the conclusion is consistent with the conclusion that the content of the wall material is consistent when the stearic acid is used as the wall material alone in part 1) (the rumen-bypass rate is affected but is not always increased along with the increase of the content).
TABLE 6 rumen bypass Rate (%)
Additive amount 2h 4h 8h 12h
1:0.5 95.47 94.36 92.73 90.76
1:1 99.13 98.74 97.03 96.47
1:1.5 99.09 98.96 96.93 96.01
1:2 99.23 98.77 97.62 96.41
1:3 99.22 98.65 97.66 96.03
4) Small intestine release rate verification
The rumen-bypass melatonin with good effect has higher rumen-bypass rate and also has higher small intestine release rate, so that the effect of protecting the melatonin from being degraded by rumen and promoting the small intestine to absorb in time can be achieved.
The rumen-bypass melatonin samples with the mass ratio of melatonin to wall of 1:1 and 1:1.5 and the mass ratio of stearic acid to hypromellose of 2:1 in 3) were selected to detect the release rate of the small intestine, and the results (table 7 below) show that the above samples have a high rumen-bypass rate but a low release rate in the small intestine. Considering the difference of the sensitivity of different wall materials to the pH value, the small intestine release rate of the 2) part stearic acid and the hydroxypropyl methylcellulose in different proportions is further verified, namely, the small intestine release rate of the stearic acid and the hydroxypropyl methylcellulose prepared when the ratio of the stearic acid to the hydroxypropyl methylcellulose is 2:0.5, 2:1 and 2:1.5 is shown by the result (table 8 below) that the small intestine release rate of the rumen-passing melatonin prepared according to the conditions that the ratio of the melatonin to the wall materials is 1:1 and 1:1.5 has no obvious change along with the increase of the content of the wall materials under the condition that the ratio of the stearic acid to the hydroxypropyl methylcellulose is the same, the small intestine release rate of the same condition that the ratio of the melatonin to the wall materials is the same has obvious change along with the change of the ratio of the stearic acid to the hydroxypropyl methylcellulose in the wall materials, and the specific small intestine release rate is obviously increased along with the use amount of the hydroxypropyl methylcellulose, so that the content of the hydroxypropyl methylcellulose has obvious positive correlation with the small intestine release rate is indicated. Further verifying the relation between the content of the hypromellose and the release rate of the small intestine, selecting stearic acid and the hypromellose as 1:2, 1:2.5, 1:3, 1:3.5 and 1:4, and performing effect verification that the mass ratio of the melatonin to the wall material is 1:1, wherein the result (table 9 below) shows that the release rate of the small intestine is obviously improved along with the increase of the content of the hypromellose, when the mass ratio of the stearic acid to the hypromellose in the wall material is 1:2-1:3, the release rate of the small intestine is basically completely released within 2 hours, and the effect of improving the content of the hypromellose is not obvious any more; in combination with the above test process, the rumen bypass rate of the rumen bypass melatonin is high when the stearic acid content is higher than that of hypromellose, but the small intestine release rate is lower; the rumen bypass rate of the rumen bypass melatonin is lower when the stearic acid content is lower than that of the hydroxypropyl methylcellulose, but the small intestine release rate is higher, as the rumen environment of the ruminant and the small intestine environment are obviously different, the pH value difference is huge, the rumen bypass melatonin firstly passes through the rumen and then reaches the small intestine after being eaten by the ruminant, the rumen bypass melatonin is suggested to be designed into a wall material with high rumen bypass rate on the outer layer and a wall material with high small intestine release rate on the inner layer during manufacturing, and the two wall materials are matched to protect the melatonin, so that the rumen bypass can be better and intestinal absorption of the co-animal can be effectively released.
TABLE 7 Small intestine Release Rate (%)
TABLE 8 Small intestine Release Rate (%)
TABLE 9 Small intestine Release Rate (%)
5) Screening of the dosage of two wall materials
The experiment shows that when the mass ratio of stearic acid to hypromellose is 2:1-3:1, the rumen bypass rate is high, and the stearic acid and hypromellose are used as an outer wall material; when the mass ratio of stearic acid to hypromellose is 1:2-1:3, the release rate of the small intestine is high, and the small intestine is used as an inner wall material; when the mass ratio of melatonin to wall material is 1:1, the rumen bypass rate and the small intestine release rate of the wall material are both higher, the following design test is used for screening the proper proportion of two layers of wall materials, the mass ratio of melatonin to inner layer wall material and outer layer wall material is 1:0.5:0.5, 1:1:1, 1:0.5:1 and 1:1:0.5, the mass ratio of stearic acid to hypromellose of the inner layer wall material is 1:2, the mass ratio of stearic acid to hypromellose of the outer layer wall material is 2:1, and the results (table 10 and table 11 below) show that when the mass ratio of melatonin to inner layer wall material and outer layer wall material is 1:1:1 or 1:0.5:1, the rumen bypass rate is the highest, the property is the most stable, and the mass ratio of the outer layer wall material to melatonin is the same under the two proportions; the release rate of the small intestine is highest when the mass ratio of melatonin to the inner wall material to the outer wall material is 1:1:0.5, and then is 1:0.5:0.5, the mass ratio of melatonin to the outer wall material is the same under the two proportion conditions, and the small intestine release rate verification test shows that the rumen-passing melatonin is directly placed in a small intestine simulation environment and is not subjected to rumen environment treatment, so that the release rate of the outer wall material is greatly influenced, the release rate of the small intestine is increased along with the reduction of the content of the outer wall material when the content of the outer wall material is low for 2 hours, the release rate of the small intestine is rapidly increased along with the increase of the content of the small intestine when the content of the outer wall material is low under the same wall material proportion condition, the results of the table 10 and the table 11 are synthesized, and finally, the optimal proportion is selected when the mass ratio of the melatonin, the inner wall material and the outer wall material is about 1:1:1 is selected, the inner wall material is preferably stearic acid, the hydroxypropyl methylcellulose is 1:2-1:3, and the outer wall material is preferably stearic acid, and the hydroxypropyl methylcellulose is preferably the hydroxypropyl methylcellulose is 1:2-1:3.
Table 10 rumen bypass ratio (%)
TABLE 11 Small intestine Release Rate (%)
Proportion of 2h 4h 8h 12h
1:0.5:0.5 88.35 90.38 99.93 100.00
1:1:1 87.38 89.13 97.38 100.00
1:0.5:1 87.42 90.03 98.05 100.00
1:1:0.5 91.45 94.83 100.00 100.00
Test example 3 practical application test
In order to verify the product performance of the rumen-protected melatonin prepared by the invention, the rumen-protected melatonin prepared by the invention in example 1, example 3 and example 5 is fed to cows, and the following specific steps are as follows:
1. 40 Holstein cows with average number of fetuses, close lactation days and higher somatic cells (the somatic cells are 40-50 ten thousand/ml) are selected, and are divided into 5 groups by adopting a single-factor test design and a random group method, wherein the number of the control groups and the number of the test groups are respectively 2, and 8 are each. Wherein, the 1 st group is only fed with basic ration, and is a control group 1; group 2 control group 2 with uncoated melatonin 18mg (daily per cow) added to the basal diet; 400mg of rumen-bypass melatonin prepared in example 1, example 2 and example 3, test group 1, test group 2 and test group 3, and 14 per day were added to each cow in groups 3 to 5 each day: 00 oral cavity feeding is carried out once, and the feeding time is 28d.
Table 4 average number of days of lactation in test cattle
Group of Control group 1 Control group 2 Test group 1 Test group 2 Test group 3
Average days of lactation 171.48 172.31 172.33 171.76 173.41
2. Test design
After the above grouping, 1 group is randomly selected from the above groupings to be used as a control group 1, the uncoated melatonin is fed to the 1 group, and the rumen-protected melatonin prepared by the invention is fed to the 3 groups of oral cavities.
Daily ration and nutrition management
During the test period, the test cows and normal lactating cows are all fed in the same colony house, milking for 3 times per day, and freely taking food and drinking water, and freely moving in the colony house, and cleaning manure at regular time per day, so that the colony house is kept clean. Feeding for 3 times a day, wherein the feeding time is respectively 7: 00. 14: 00. 19:00. the test animals were free to eat and drink water, and the daily ration ingredients and feed ingredients of the test animals during the test period were as shown in tables 5 and 6 below.
TABLE 5 cow ration composition (air dried basis) and nutrient level (DM basis)
Note that: in Table 5
1) The main component of the beauty force is rumen bypass fatty acid calcium.
2) Each kilogram of premix comprises: stone powder 426.633g, baking soda 225g, table salt 25g, rumen bypass fat 25g, methionine 8.75g, selenium enrichment 7.5g, manganese 3.313g, copper 2.75g, calcium 97.481g, cobalt 1.75g, zinc 8.188g, magnesium 87.5g, biotin 1.406g, VA 0.275g, VD 3.11 g and VE 3.094g.
3) The nutrient level was found. The levels of nutrients such as dry matter, crude protein, crude fat, crude fiber, etc. were determined with reference to the international method.
Table 6 nutritional composition and content of feed raw materials (DM basis)
2. Statistical analysis and test results
The statistical method comprises the following steps: data analysis was performed using SPSS 19.0.
TABLE 7 average bovine milk cell number (ten thousand/ml)
Control group 1 Control group 2 Test group 1 Test group 2 Test group 3
Feeding for 7 days 41.31 40.43 25.76 24.92 23.58
Feeding for 14 days 42.53 41.35 19.38 19.67 18.93
Feeding for 21 days 41.92 41.78 16.63 17.14 15.78
Feeding for 28 days 41.66 41.13 17.19 17.47 15.44
Table 8 test cow milk protein content average (%)
Control group 1 Control group 2 Test group 1 Test group 2 Test group 3
Feeding for 7 days 3.10 3.14 4.12 4.09 4.18
Feeding for 14 days 3.14 3.26 4.33 4.23 4.26
Feeding for 21 days 3.09 3.32 4.41 4.39 4.35
Feeding for 28 days 3.04 3.38 4.16 4.40 4.37
Table 9 test average daily milk yield (kg)
Control group 1 Control group 2 Test group 1 Test group 2 Test group 3
Feeding for 7 days 18.71 18.82 21.36 22.19 21.79
Feeding for 14 days 18.63 18.79 21.79 22.32 22.31
Feeding for 21 days 18.36 19.12 22.32 22.41 22.18
Feeding for 28 days 17.43 19.09 22.27 22.33 22.72
From the above, the rumen-bypass melatonin of the invention can effectively improve the milk quality, improve the milk protein content, reduce the number of somatic cells and improve the daily output of cow milk.
It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Claims (10)

1. The rumen-bypass melatonin product is characterized by comprising a core material, a first wall material and a second wall material, wherein the first wall material and the second wall material are sequentially coated outside the core material; the core material is melatonin, and the first wall material and the second wall material both comprise stearic acid and hypromellose.
2. The rumen-bypass melatonin product of claim 1, wherein the mass ratio of melatonin to the first wall material to the second wall material in the rumen-bypass melatonin product is 0.9-1.1:0.9-1.1:0.9-1.1.
3. The rumen-bypass melatonin product of claim 1 or 2, wherein the mass ratio of stearic acid to hypromellose in the first wall material is 1:2-3, and the mass ratio of stearic acid to hypromellose in the second wall material is 2-3:1.
4. A method for preparing the rumen-bypass melatonin product of claims 1-3, comprising the steps of:
s1, taking melatonin, adding a proper amount of water, uniformly mixing, preparing into strip-shaped particles in a granulator, transferring into a shot blasting machine, rotating, boiling and blasting, and drying to obtain melatonin pellets; transferring melatonin pellets into a fluidized bed for fluidization treatment to obtain fluidized pellets;
s2, heating the first wall material to 120 ℃ for melting, cooling to 80-100 ℃, and coating the first wall material on the surface of the fluidized pellets to obtain rumen melatonin intermediates;
and S3, heating the second wall material to 120 ℃ for melting, cooling to 80-100 ℃, and coating the second wall material on the surface of the rumen-bypass melatonin intermediate to obtain the rumen-bypass melatonin product.
5. The method for preparing a rumen-bypass melatonin product according to claim 4, wherein the moisture content of the melatonin pellets after drying in step S1 is less than or equal to 10wt%.
6. The method for preparing the rumen-bypass melatonin according to claim 4, wherein the atomizing pressure of the fluidized bed in the step S1 is 3.5kg/cm 2, the rotating speed is 90-95r/min, the air inlet temperature is 35-45 ℃, and the air outlet temperature is 25-45 ℃.
7. The use of the rumen-bypass melatonin product of claims 1-6, for feeding cows, thereby reducing the number of somatic cells in cows ' milk, increasing the protein content in cows ' milk, and increasing the milk production of cows ' milk.
8. The use according to claim 7, wherein the breed of cows is a holstein cow.
9. The use according to claim 7, wherein 400-450mg rumen by-pass melatonin product is fed per cow per day.
10. The use according to claim 7, wherein after feeding cows with the rumen-protected melatonin product, the milk protein content of cows is increased by 30-40%, the number of somatic cells is reduced by 60-70% and the milk yield is increased by 10-15% compared to cows not fed with the rumen-protected melatonin product.
CN202410398999.3A 2024-04-03 2024-04-03 Rumen-bypass melatonin product and preparation method and application thereof Pending CN118160831A (en)

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