CN111100039A

CN111100039A - Preparation method and application of argininyl zinc

Info

Publication number: CN111100039A
Application number: CN201911348855.2A
Authority: CN
Inventors: 刘伟; 黄逸强; 邓敏; 王赏初; 陈光伟
Original assignee: Changsha Xingjia Biological Engineering Co Ltd
Current assignee: Changsha Xingjia Biological Engineering Co Ltd
Priority date: 2019-12-24
Filing date: 2019-12-24
Publication date: 2020-05-05

Abstract

The invention discloses a preparation method and application of arginine biotin zinc, wherein the preparation method comprises the following steps: reacting arginine biotin with carbonate or bicarbonate in water, stirring until no air bubbles emerge, heating, and adding inorganic zinc salt to react for a period of time to obtain arginine biotin zinc. The prepared arginine biotin zinc is applied as an animal feed additive, and the animal is a pig, a bird, a ruminant or an aquatic animal. The preparation method of the arginine biotin zinc has the advantages of simple process, low cost, less energy consumption, higher yield and high product quality. The argininyl zinc has the advantages of high biological value, low addition amount, obvious growth promoting effect, less side effect of animals, long-term use, safety, no drug resistance and the like.

Description

Preparation method and application of argininyl zinc

Technical Field

The invention relates to preparation and application of a feed additive, in particular to application of an amino acid complex as the feed additive and a preparation method thereof.

Background

The arginine biotin can participate in the urea circulation of the organism in the animal body, has long half-life period and stable metabolism, and has wide biological functions. The research shows that arginine biotin can be used as a metabolic activator to participate in the activation of dihydropyrrole-5-carboxylic acid synthetase (P5CS) and carbamyl phosphate synthetase I (CPS-I), promotes the synthesis of citrulline from glutamine or proline, and further promotes the synthesis of arginine, so that the arginine biotin is also called as arginine endogenous activator. Nitric oxide and polyamines (including putrescine, spermidine and spermine) have a wide range of actions in the body, and can promote local blood circulation, regulate and control the nutritional metabolism and immune response of the body. The arginins can also have significant effects on multiparous animals such as pigs, dogs, rabbits and the like, and can improve litter size and birth weight of female animals.

Arginine is indispensable as an essential precursor for synthesizing the arginine, but the cost of adding effective dose of arginine in feed is too high, and other amino acids such as lysine and tryptophan have antagonistic effect, so the application in animal production is greatly limited. The argininol can effectively increase the content of arginine in animals, and is very important for maintaining the arginine level and the normal physiological function of organisms. The research of the applicant shows that compared with the direct use of arginine, the use of arginine in animal production has the following advantages: 1) comparison of effective dose versus cost: tests on sows and piglets show that the same effect can be achieved by adding about 1/10 parts of arginine in the arginine biotin, and the cost is obviously superior; 2) comparison of metabolic processes in vivo: arginine is very easy to hydrolyze by arginase in animals and cannot maintain a higher concentration level, while arginine biotin is stably metabolized in the bodies, and the production of arginine is continuously promoted through a urea circulation passage, so that the arginine of the bodies can be maintained at a certain physiological concentration level for a long time; 3) influence on the amino acid balance: arginine with drug dosage is required to be added to play the reproductive potential of the multiparous animals, but the addition of excessively high arginine in daily ration can interfere the balance of amino acid, and a small amount of arginine biotin can be added to synthesize in the organism and maintain a higher arginine level, thereby playing the important function of the multiparous animals and avoiding the antagonism with lysine and the like; 4) effect on other amino acid uptake: oral administration of arginine to animals interferes with the absorption of lysine, histidine and tryptophan in the small intestine, resulting in a decrease in the plasma concentration of these amino acids; the arginine biotin does not interfere with the absorption of the amino acids and does not influence the concentration of the arginine biotin in blood plasma, thereby playing a role in saving essential amino acids to a certain extent; 5) storage and ease of use comparison: the natural active L-arginine is white powder, but is easy to absorb moisture and difficult to store, so that L-arginine hydrochloride is mostly used in the production, the chlorine content of the product is 16.8-17%, chloride ions can be increased when the L-arginine hydrochloride is added into animal ration, the electrolyte balance of an organism is influenced, arginine is stable in physicochemical properties at normal temperature and normal pressure, the L-arginine hydrochloride can be stored for a long time, and the L-arginine hydrochloride cannot interfere with the electrolyte balance when the L-arginine hydrochloride is added into the animal ration.

Since the first confirmation of zinc as one of the essential trace elements for animal nutrition by rat test in 1934 by Todd et al, zinc is a component of more than 200 kinds of metalloenzymes, hormones and insulin in animal organism, and has been gradually found to promote growth and tissue regeneration of organism, maintain normal metabolism of organism, promote appetite, maintain normal sexual organs and functions, accelerate wound healing, protect skin health, enhance immune mechanism and improve resistance.

The preparation of zinc arginate mentioned in the Synthesis, Crystal Structure and Spectroscopic Properties of Zn (II) with N-carbamyglutamate Ligand by SHU Xu-Gang et al is carried out by reacting zinc oxide with arginate. The preparation method has certain defects that zinc oxide is insoluble, and arginine biotin can dissolve the zinc oxide to release zinc ions only by releasing hydrogen ions with certain concentration, and then the zinc ions react with the zinc ions to generate the arginine biotin zinc. When the concentration of the arginine biotin is reduced to a certain value, the concentration of ionized hydrogen ions is reduced, the zinc oxide is difficult to dissolve again, the reaction is incomplete, the zinc oxide remained in the product cannot be removed by the subsequent process, and the yield and the purity of the product are reduced. Similarly, the same applies to insoluble alkaline materials.

Disclosure of Invention

The invention aims to overcome the defects of the prior art, provides a preparation method of arginine biotin zinc with simple process, low cost, low energy consumption, higher yield and high product quality, and also provides application of the arginine biotin zinc as an animal feed additive, wherein the arginine biotin zinc has the advantages of high biological value, low addition amount, obvious growth promoting effect, few animal side effects, long-term use, safety and no drug resistance.

In order to solve the technical problems, the invention also provides a preparation method of the arginine biotin zinc used as the animal feed additive, which comprises the following steps: reacting arginine biotin with carbonate or bicarbonate in water, stirring until no air bubbles emerge, heating, and adding inorganic zinc salt to react for a period of time to obtain arginine biotin zinc. Specifically, the arginine biotin can be added into water, stirring is started, carbonate or bicarbonate is added, stirring is carried out until no bubbles emerge, then the temperature is raised, inorganic zinc salt is added, reaction is carried out for a period of time, and after the reaction is finished, the product is obtained through post-treatment.

In the above preparation method, preferably, the carbonate is sodium carbonate and/or potassium carbonate, and the bicarbonate is sodium bicarbonate and/or potassium bicarbonate.

In the preparation method, the mol ratio of the carbonate to the argininyl is preferably (1.01-1.02) to 1; the molar ratio of the bicarbonate to the argininyl is (2.02-2.04) to 1.

The arginino is affected by its structure, and under the action of high temp. and strong alkali (pH value is greater than 12), the-NHCO on arginino^-Hydrolysis reaction to form-NH may occur₂and-COO^-. In order to avoid the reaction as much as possible, the arginine biotin is firstly reacted with the carbonate or bicarbonate with weaker basicity at normal temperature. Since arginins react mildly with strong bases and weak acid salts and are exothermic, no heating is required. As the reaction proceeds, the carbonate or bicarbonate is continuously consumed, the generated carbon dioxide is continuously escaped, and the pH value is gradually changed from alkalinity to alkalescence. However, at pH < 7.5, the arginine biotin may undergo intermolecular and intramolecular dehydration condensation reactions. Therefore, in the present invention, although the adjustment of the pH is not emphasized, a slight excess of the base is required to reduce the occurrence of side reactions. However, if the carbonate or bicarbonate is too excessive, the alkaline concentration in the solution is too high, and the carbonate formed by the escaped carbon dioxide dissolved in water can generate carbonate ions in the solution with higher pH, and the carbonate ions react with the subsequent zinc ions to generate zinc carbonate, which is not beneficial to the subsequent reaction, so the molar ratio of the carbonate or bicarbonate to the arginino is selected from the above range.

In addition, the carbonate or bicarbonate does not generate decomposition reaction by itself in a lower-temperature aqueous solution to generate carbon dioxide to escape, so the bubble emission is a byproduct generated by the reaction of the carbonate or bicarbonate and the arginine biotin. Stirring until no air bubbles are emitted, and then considering that the arginine biotin and the carbonate or bicarbonate have basically completely reacted, even if the arginine biotin and the carbonate or bicarbonate are not completely reacted, a small amount of residual arginine biotin and carbonate or bicarbonate can continue to react, and the subsequent reaction is not influenced.

In the above preparation method, preferably, the inorganic zinc salt includes zinc chloride and/or zinc sulfate. Zinc sulfate also includes forms containing crystalline water such as zinc sulfate monohydrate.

In the above preparation method, the molar ratio of the inorganic zinc salt to the argininyl is preferably (0.97-1.03): 1.

In the above production method, it is preferable that the reaction temperature is not more than 60 ℃. The reaction temperature is too high, and side reactions such as condensation and the like of arginine biotin salt can occur, so the reaction temperature cannot be too high.

In the above production method, the reaction time is preferably 1.1 to 2 hours. In the preparation method, the time is too short, incomplete reaction is easily caused, and the product conversion rate is low; the reaction conversion rate is not obviously increased after the reaction time is too long, and part of water is evaporated, so that the material is too viscous and agglomerated, the crystallization granularity is influenced, and the discharging and centrifuging are not facilitated. It is also considered that even if the solution contains a trace amount of zinc carbonate, zinc argininate can be produced by reaction with the argininyl salt. The proper reaction time can ensure complete reaction and higher yield, and reduce the raw material waste caused by insufficient reaction time and the energy consumption caused by long-time reaction.

In the above preparation method, preferably, the solid content of the reaction system is 18 wt% to 45 wt%.

In the preparation method, when the solid content of the reaction system is 18-28%, after the reaction is finished, an organic solvent is added, the mixture is filtered and dried, the organic solvent is one or more of ethanol, propanol and isopropanol, and the volume ratio of the organic solvent to the reaction solution is 0.05-0.2: 1.

Because the solubility of the argininyl zinc in water is lower than that of sodium salt/potassium salt, the argininyl zinc can be separated out firstly under the condition of proper solid content. The solid content is too high, and partial sodium salt/potassium salt can be separated out while arginine biotin zinc is separated out, so that co-crystal is formed.

The solid content of the reaction system can be divided into two gradients, wherein the first gradient is low solid content, and the solid content is 18-28%; the second gradient is high solid content, with a solid content of less than 45% and greater than 28%. The preparation method with low solid content adopts the subsequent process of adding organic solvent for crystallization and then filtering and drying, and is suitable for high-grade monomer products with higher purity. The preparation method with high solid content adopts a direct concentration spray drying method subsequently, the yield is close to 100%, the product contains by-product sodium salt/potassium salt, and the product is mainly a medium-grade product with lower purity and is even used in premixed feed. In addition, the preparation method with high solid content is suitable for areas with strict environmental protection control, does not need to discharge salt-containing wastewater, or treats the salt-containing wastewater after concentration and drying, and saves the cost.

In the invention, arginine biotin firstly reacts with carbonate or bicarbonate to generate arginine biotin salt, and then reacts with inorganic zinc salt under the heating condition to generate arginine biotin zinc (C)₆H₈N₂O₅Zn·H₂O), the chemical reaction equation is as follows:

the first step is as follows:

C₆H₁₀N₂O₅+Na₂CO₃＝C₆H₈N₂O₅Na₂·H₂O+CO₂×) ×; or

C₆H₁₀N₂O₅+2NaHCO₃＝C₆H₈N₂O₅Na₂·H₂O+2CO₂×) ×; or

C₆H₁₀N₂O₅+K₂CO₃＝C₆H₈N₂O₅K₂·H₂O+CO₂×) ×; or

C₆H₁₀N₂O₅+2KHCO₃＝C₆H₈N₂O₅K₂·H₂O+2CO₂↑；

The second step is that:

C₆H₈N₂O₅Na₂·H₂O+ZnSO₄＝C₆H₈N₂O₅Zn·H₂O↓+Na₂SO₄(ii) a Or

C₆H₈N₂O₅Na₂·H₂O+ZnCl₂＝C₆H₈N₂O₅Zn·H₂O ↓ +2 NaCl; or

C₆H₈N₂O₅K₂·H₂O+ZnSO₄＝C₆H₈N₂O₅Zn·H₂O↓+K₂SO₄(ii) a Or

C₆H₈N₂O₅K₂·H₂O+ZnCl₂＝C₆H₈N₂O₅Zn·H₂O↓+2KCl。

In the above method, the reaction principle is as follows: firstly, reacting strong alkali and weak acid salt with argininyl to generate argininyl salt and carbon dioxide at normal temperature; secondly, under heating, the arginine biotin salt reacts with the zinc salt to generate arginine biotin zinc and sodium salt/potassium salt.

If argininol reacts directly with zinc salt and then strong base is added to adjust the pH value to generate precipitate to prepare argininol zinc, firstly, the reaction rate of argininol and zinc salt is very slow at low temperature, the time is too long, and the industrial production is not facilitated, so the reaction needs to be heated, and secondly, the strong base can cause-NHCO on the argininol zinc^-Hydrolysis reaction occurs, impurities increase, and the yield becomes low.

As a general technical concept, the invention also provides an application of the arginine biotin zinc prepared by the preparation method as an animal feed additive.

In the above application, the animals are pigs, birds, ruminants and aquatic animals.

In the application, preferably, the addition amount of the argininyl zinc in each ton of pig feed is 20-100 ppm calculated by zinc element; the addition amount of the zinc element in each ton of poultry feed is 20-100 ppm; the addition amount of the zinc element in each ton of aquatic feed is 20-120 ppm.

The arginine zinc with equal zinc or low zinc level is added into the feed to replace the high-dose inorganic zinc sulfate used in the conventional daily ration, and the arginine zinc with equal zinc or low zinc level is applied to the pig feed, so that the feed has the effects of obviously improving the reproductive performance of sows, improving the intestinal health of weaned piglets, promoting growth and improving the immunity of the pigs at each stage; the feed has the effects of obviously improving the meat quality of meat poultry, promoting the growth, improving the antibody level, improving the eggshell quality and improving the breeding of breeding poultry and the hatching performance of hatching eggs; the composition is applied to aquatic animals, and has the effects of improving oxidation resistance and immunity and promoting growth.

Compared with the traditional method, the preparation method of the arginine zinc has the advantages of simple process, low cost, less energy consumption, higher yield and high product quality. In particular, the present invention has the following advantages:

(1) the carbonate or bicarbonate and zinc salt added during the reaction are soluble, the argininol is also soluble, and even if the raw materials are not reacted completely, the argininol can be dissolved in the aqueous solution and cannot be precipitated in the product to cause the reduction of the product purity.

(2) The reaction product of the invention is soluble in water, but insoluble or slightly soluble in organic solvent, so that better crystallization can be realized by adding the organic solvent, the precipitated product particles are more dispersed, the crystal form is better, and the crystallized product obtained after filtration has less impurities, higher purity and higher yield.

(3) The reaction only needs to calculate the weight of the added carbonate or bicarbonate, does not need to adjust the pH value in the reaction, and reduces the workload.

When the arginine biotin zinc prepared by the method is used as an animal feed additive to be applied, the arginine biotin zinc does not generate antagonistic reaction and redox reaction with other components in the feed, and the stability, high quality and high efficiency of the feed can be ensured; and can enter the body in ways of endocytosis, active transportation and the like, has small stimulation to intestines and stomach, and can ensure that the animal body can simultaneously and better absorb arginine biotin and zinc. The zinc L-arginine can also obviously improve the utilization rate of elements, greatly reduce the discharge amount of zinc elements in animal excreta and effectively reduce the pollution to the environment. The argininyl zinc has the advantages of high biological value, low addition amount, obvious growth promoting effect, less side effect of animals, long-term use, safety, no drug resistance and the like.

Detailed Description

The invention is further described with reference to specific examples.

In order to facilitate an understanding of the present invention, the present invention will be described more fully and in detail with reference to the preferred embodiments, but the scope of the present invention is not limited to the specific embodiments below.

Unless otherwise defined, all terms of art used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention.

Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by existing methods.

The zinc content of the product of the following examples was determined by xylenol orange EDTA titration, arginine biotin was determined by azotometry, sodium was determined by atomic absorption spectrophotometer, potassium was determined by atomic absorption spectrophotometer, chloride ion was determined by silver nitrate titration, sulfate radical was determined by barium chloride precipitation, water loss at 180 ℃ was lost as lost crystal water and at 104 ℃ was lost as lost free water.

Example 1: preparation of argininyl zinc

Adding 388Kg of arginine biotin with the purity of 98 percent into 2.39t of water, starting stirring, then adding 218.5Kg of sodium carbonate with the purity of 98 percent, stirring until no air bubbles emerge, heating to 40 ℃, then adding 369.9Kg of zinc sulfate monohydrate with the purity of 98 percent, reacting for 2h, concentrating and spray-drying after the reaction is finished, and obtaining 853.6Kg of a product.

The content of argininol in the argininol zinc product is 43.9 percent by measurement, and Zn is obtained²⁺15.3% of Na⁺10.7% SO₄ ^2-22.3%, the water loss at 180 ℃ is 4.2%, the water loss at 104 ℃ is 2.3%, i.e. the purity is 96.4%, the yield is 99.7% calculated by arginine biotin, i.e. the molar ratio of arginine biotin to zinc is about 1:1, the molecular formula is C₆H₈N₂O₅Zn·H₂O。

Example 2: preparation of argininyl zinc

Adding 388Kg of arginine biotin with the purity of 98 percent into 1.5t of water, starting stirring, then adding 416.8Kg of potassium bicarbonate with the purity of 98 percent, stirring until no air bubbles appear, heating to 30 ℃, then adding 598.6Kg of zinc sulfate heptahydrate with the purity of 98 percent, reacting for 1.1h, concentrating and spray-drying after the reaction is finished, and obtaining 560.0Kg of a product.

The content of the argininol in the argininol zinc product is measured to be 40.7 percent, and Zn is measured²⁺14.2% by weight, K⁺17.1% SO₄ ^2-20.9%, the water loss rate at 180 ℃ is 3.9%, the water loss rate at 104 ℃ is 2.3%, namely the purity is 96.8%, the yield is 99.8% calculated by arginine biotin, namely the molar ratio of arginine biotin to zinc is about 1:1, and the molecular formula is C₆H₈N₂O₅Zn·H₂O。

Example 3: preparation of argininyl zinc

Adding 388Kg of arginine biotin with the purity of 98 percent into 4.6t of water, starting stirring, adding 346.3Kg of sodium bicarbonate with the purity of 98 percent, stirring until no bubbles emerge, heating to 60 ℃, adding 278.2Kg of zinc chloride with the purity of 98 percent, reacting for 1.4h, cooling to below 40 ℃ after the reaction is finished, adding 920L of ethanol, crystallizing, filter-pressing, flash evaporating and drying to obtain 511.5Kg of a product.

The content of argininol in the argininol zinc product is 68.0 percent and Zn is measured²⁺23.6%, the water loss at 180 ℃ is 6.5%, the water loss at 104 ℃ is 1.3%, i.e. the purity is 98.1%, the yield is 92.4% calculated by arginine biotin, i.e. the molar ratio of arginine biotin to zinc is about 1:1, the molecular formula is C₆H₈N₂O₅Zn·H₂O。

Example 4: preparation of argininyl zinc

Adding 388Kg of arginine biotin with the purity of 98 percent into 2.5t of water, starting stirring, adding 287.7Kg of potassium carbonate with the purity of 98 percent, stirring until no bubbles emerge, heating to 50 ℃, adding 272.6Kg of zinc chloride with the purity of 98 percent, reacting for 1.6h, cooling to below 40 ℃ after the reaction is finished, adding 125L of propanol, crystallizing, filter-pressing, flash evaporating and drying to obtain 511.7Kg of product.

Measuring arginine in the arginine biotin zinc productBiotin content of 68.2%, Zn²⁺23.7%, the water loss at 180 ℃ is 6.5%, the water loss at 104 ℃ is 1.3%, i.e. the purity is 98.4%, the yield is 92.7% calculated by arginine biotin, i.e. the molar ratio of arginine biotin to zinc is about 1:1, the molecular formula is C₆H₈N₂O₅Zn·H₂O。

Application example 1: the application of the argininyl zinc as a feed additive for sows.

The zinc argininate prepared in example 3 was used as a feed additive for feeding sows.

The test selects healthy long x sow with pregnancy (106 +/-1) d and similar weight and gestation times, 40 sows are randomly divided into 5 groups, each group has 8 repetitions, and each repetition has 1 repetition. Zinc sulfate (100ppm in terms of zinc) was added to the basal diet in 1 group, and arginine biotin zinc (20ppm in terms of zinc), arginine biotin zinc (40ppm in terms of zinc), arginine biotin zinc (60ppm in terms of zinc), and arginine biotin zinc (100ppm in terms of zinc) were added to the basal diet in 2, 3, 4, and 5 groups, respectively. The basal ration was formulated with reference to the NRC (2102) standard. The feeding mode and daily management procedures are normally carried out according to the pig farm regulations.

Index detection: (1) production achievement: calculating litter weight, average weaning weight, average 21d weight gain and daily gain of the weaned piglets. The days between weaning and re-estrus of the sows were recorded. (2) Blood index: collecting 10ml of blood from ear vein during production of sow, anticoagulating with heparin sodium, standing for 20min, centrifuging at 3000r/min for 15min, taking supernatant EP tube, subpackaging, and immediately placing in-20 deg.C refrigerator for storage. The immunoglobulin G (IgG) and the immunoglobulin M (IgM) of the plasma of the sow are measured by a Beckmann CX4 full-automatic biochemical analyzer. The kit is provided by Beijing Lidman Biochemical technology, Inc. The test results are shown in Table 1.

TABLE 1 Effect of different treatments on sow Productivity and serum Immunity index

Item	Group 1	2 groups of	Group 3	4 groups of	5 groups of
						Weight of weaning litter (kg)	48.1±4.5^b	49.2±9.7^b	53.4±6.7^b	53.9±7.8^b	57.2±7.7^a
Weight average weaning (kg)	5.3±0.6	5.7±0.8	5.9±0.6	6.0±0.5	5.9±0.4
						Daily gain (g/d)	179.3±18.3^b	182.8±25.9^b	204.8±29.9^ab	213.0±12.8^a	206.6±11.4^ab
Estrus interval (d)	4.8±1.5	5.3±1.0	5.3±1.0	5.6±1.1	5.6±1.4
						IgG(mg/dl)	279.62±30.24^b	306.90±27.47^ab	309.89±18.93^ab	322.96±29.96^a	324.45±24.66^a
IgM(mg/dl)	20.55±6.55	23.56±5.78	23.46±5.32	25.61±2.55	24.09±3.83

Note: the case where the letters in the same row are completely different indicates that the difference is significant (P < 0.05).

As can be seen from the above table 1, the average weight of weaning litter and the daily gain tend to increase with the increase of the addition amount of zinc argininate in the sow ration. Wherein the weight of the weaned litter of the sow fed with the additive argininol zinc (80ppm calculated by zinc) is the highest and is obviously higher than that of the sow fed with the additive zinc sulfate (80ppm calculated by zinc). The daily gain of the suckling piglets fed with the zinc L-arginine (60ppm calculated by zinc) is the highest and is obviously higher than that of the control group fed with the zinc sulfate (80ppm calculated by zinc). The serum IgG of the sows fed with the additive argininyl zinc (80ppm calculated by zinc) and argininyl zinc (60ppm calculated by zinc) is obviously higher than that of the group 1. There was no significant difference from the other groups. The fact that a certain amount of arginine zinc is added can improve the production performance of sows and enhance the immunity.

Application example 2: the application of arginine biotin zinc as a weaned pig feed additive.

The zinc argininate prepared in example 3 was used as a feed additive for feeding weaned piglets.

200 28-day-old weaned three-element piglets 'Du multiplied by long multiplied by big' are selected in the test, and are randomly divided into 5 groups according to the principle that the weights (about 7.73 kg) are similar and the proportion of male to female is basically consistent, each group has 5 repetitions, and each repetition has 8 repetitions. Zinc sulfate (80ppm in terms of zinc) was added to the basal diet in 1 group, and arginine biotin zinc (20ppm in terms of zinc), arginine biotin zinc (40ppm in terms of zinc), arginine biotin zinc (60ppm in terms of zinc), and arginine biotin zinc (100ppm in terms of zinc) were added to the basal diet in 2, 3, 4, and 5 groups, respectively. The basal ration was formulated with reference to the NRC (2102) standard. The feeding mode and daily management procedures are normally carried out according to the pig farm regulations. The pilot run was 7 days and the positive run was 21 days.

Index detection: (1) and (3) measuring the growth performance: piglets were weighed on empty stomach on test day 1 and day 28 at 08:30, respectively, daily feed intake was recorded, and average daily gain and feed-weight ratio were calculated. (2) And (3) determining the intestinal structure: on the 21 st day of the experiment, 4 piglets are randomly selected from each group, killed by bloodletting through jugular vein after no feeding for 24h, the middle section of jejunum about 3cm is taken, the content is washed by normal saline, the jejunum is fixed in 4% paraformaldehyde solution, and slices are made for determining the intestinal morphological structure. Each group of slices selects 8 longest villi with complete crypt shape, and the average height, width and depth of the villi of each group are calculated. The test results are shown in Table 2.

TABLE 2 Effect of different treatments on growth Performance and intestinal morphology of weaned piglets

From the above table 2, it can be seen that the addition of zinc argininate of more than 40ppm in terms of zinc can significantly improve the average daily gain of weaned pigs, reduce the feed-weight ratio, and significantly reduce the crypt depth of jejunum and increase the villus height. The effect is optimal when 100ppm of zinc is calculated by adding arginine zinc, and the daily gain is obviously higher than that of other groups. The fact that a proper amount of arginine zinc is added into the feed of the weaned pigs to replace high-dose zinc sulfate can improve the growth performance of the weaned pigs and promote the establishment of intestinal tracts.

Application example 3: the application of the argininyl biotin zinc as a broiler breeder feed additive.

The arginine zinc prepared in example 3 was used as a feed additive for broiler breeder chickens.

The test adopts a single-factor random grouping design, 750 fast-growing south yellow-feather broiler breeder parental breeding hens with 37 weeks old and the same health and genetic background are selected as test chickens, and the test chickens are randomly divided into 5 treatment groups according to the principle that the body weight and the laying rate are basically consistent, each group is divided into 5 times, each group is divided into 15 times, and each cage is divided into 2 chickens. Zinc sulfate (100ppm in terms of zinc) was added to the basal diet in 1 group, and arginine biotin zinc (40ppm in terms of zinc), arginine biotin zinc (60ppm in terms of zinc), arginine biotin zinc (80ppm in terms of zinc), and arginine biotin zinc (100ppm in terms of zinc) were added to the basal diet in 2, 3, 4, and 5 groups, respectively. The test adopts corn-soybean meal type basic feed, and the nutrition level of the basic feed is mainly prepared according to chicken feeding standard (NY/T33-2004). Feeding in a stepped cage-feeding limited feeding mode, feeding test chickens 125 g/time every day at a ratio of 09:00, freely drinking water, and keeping constant illumination for 16 hours every day. All the chickens were prefed with the feed of group 1 for 10 days, and after the egg laying performance was stable, the test period was 10 weeks.

The indexes are determined as follows (1) egg laying performance: during the test period, eggs are picked up 1 time at 15:00 every day, the number of each repeated egg and the number of unqualified eggs are recorded, the total egg weight is weighed, and the egg laying rate, the average egg weight, the total feed consumption, the feed-egg ratio and the unqualified egg rate are calculated. (2) Eggshell quality: and (3) randomly selecting 3 qualified hatching eggs every time when the test is finished, and finishing the egg quality determination within 48 hours, wherein the egg quality determination comprises the eggshell strength and the eggshell thickness. Measuring the thicknesses of 3 points of the blunt end, the middle end and the sharp end of the eggshell after the eggshell membrane is removed by adopting a micrometer, and taking the average value to obtain an eggshell thickness value; the eggshell strength is measured by adopting an eggshell strength tester. (3) Hatching performance: and (3) repeatedly selecting 60 qualified hatching eggs 1 week before the test, marking, incubating in an incubator, and measuring the fertility rate, the hatching rate, the healthy chick rate and the average hatched weight of the healthy chick.

TABLE 3 influence of different treatments on the productivity and egg quality of yellow-feather broiler breeder breeders

As can be seen from the above Table 3, the egg laying rate and the feed conversion rate of various groups added with arginine zinc of more than 60ppm calculated by zinc are higher than those of the control group added with high-dose inorganic zinc sulfate, but no significant difference exists. The egg rate, the eggshell strength and the eggshell thickness of the arginine-biotin-zinc combined lattice added by 80ppm and 100ppm calculated by zinc are obviously higher than those of other groups.

TABLE 4 Effect of different treatments on hatching performance of yellow-feathered broiler breeder eggs

As can be seen from Table 4, the fertilization rates of the hatching eggs of the group added with 80ppm and 100ppm of zinc argininate are obviously higher than those of the control group, and the hatching egg hatching rate of the group added with 100ppm of zinc argininate is obviously higher than that of the control group. The healthy chick rate is increased along with the increase of the addition amount of the arginine biotin.

In conclusion, arginine zinc with the concentration of more than 60ppm calculated by zinc is added to the daily feed of the broiler breeder to replace 100ppm zinc sulfate with high dosage calculated by zinc, so that the broiler breeder feed has the effects of obviously improving the egg laying performance of the broiler breeder, improving the eggshell quality, reducing the unqualified egg rate and improving the hatching performance of hatching eggs.

Application example 4: the application of arginine biotin zinc as a broiler feed additive.

Selecting 900 feather young chickens which are hatched from the same hatchery and are healthy and have similar physical quality and age of 1 day. A single-factor completely random design is adopted, and the random division is carried out into 5 groups, each treatment group has 6 repetitions, and each repetition has 30 feathers. Wherein zinc sulfate (90ppm in terms of zinc) is added to the basic ration in 1 group, and arginine zinc (20ppm in terms of zinc), arginine zinc (50ppm in terms of zinc), arginine zinc (70ppm in terms of zinc), and arginine zinc (90ppm in terms of zinc) are added to the basic ration in 2, 3, 4, and 5 groups, respectively. Refer to broiler chicken nutritional requirements design formula recommended by U.S. NRC (1994) and NY/T33-2004). The test period was 42 days. The broiler chickens are placed in the henhouse in a semi-open type, are raised in 2 layers of cages and are fed with water and water freely. Immunization was performed according to conventional immunization protocols. Daily management is carried out according to the manual of AA broiler chicken feeding management.

The detection indexes comprise (1) growth performance measurement, namely weighing all chickens after fasting for 12 hours at 1 day age and 42 days age, counting feed intake, calculating a feed consumption and weight gain ratio, (2) measurement of oxidation resistance and antibody titer, wherein at 42 days age, 2 chickens are randomly drawn for each repetition and subjected to wing vein blood collection to prepare serum, the serum is stored at 20 ℃ for later use, the kit is used for measuring the concentration of Malondialdehyde (MDA) and the activities of glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD), the kit is purchased from Nanjing Biotechnology limited, and the chicken newcastle disease antibody titer is measured by using β -trace hemagglutination inhibition method, (3) meat quality measurement, namely, right-side pectoral muscles are taken after slaughtering, placed in sealing plastic after weighing (W1), and are placed in a refrigerator at 4 ℃ for 24 hours, wiped by using filter paper and weighed (W2), drip loss (W1-W2)/W1%, and the following table 5 shows the results.

TABLE 5 Effect of different treatments on the growth Performance of AA broilers

Remarking: different lower case letters in the same row indicate significant difference (P <0.05), and different upper case letters indicate significant difference (P < 0.01). The following table is the same.

As can be seen from table 5 above, the average daily gain weight of the broilers of 3, 4 and 5 groups added with 50ppm, 70ppm and 90ppm arginino zinc calculated as zinc was significantly higher than that of 1 group added with high dose of 90ppm zinc sulfate calculated as zinc. The weight gain ratio of broiler feeds in groups 4 and 5 to which 70ppm and 90ppm arginine biotin zinc calculated as zinc were added was significantly lower than that in group 1 to which 90ppm zinc sulfate calculated as zinc was added at a high dose. With different treatments, the growth performance of the broiler chicken shows an increasing trend along with the increase of the addition amount of arginine biotin zinc.

TABLE 6 Effect of different treatments on AA broiler serum antioxidant enzyme Activity and Newcastle disease titer

As can be seen from the above Table 6, the total superoxide dismutase and glutathione peroxidase of the broilers in the group of 70ppm and 90ppm arginine, zinc and the antibody titer of Newcastle disease added are both significantly higher than that in the group of zinc sulfate added with high dose, 90ppm zinc. The indexes among other groups have no obvious difference.

Table 7 effect of different treatments on meat quality of AA broilers

As can be seen from table 7 above, the loss of water dripping from the dorsum muscles of the broilers of groups 3, 4 and 5 to which 50ppm, 70ppm and 90ppm argininyl zinc, calculated as zinc, was added was significantly lower than that of group 1 to which 90ppm zinc sulfate, calculated as zinc, was added at a high dose and group 2 to which 20ppm argininyl zinc, calculated as zinc, was added at a low dose.

In conclusion, the addition of a proper amount of argininol in the conventional basic ration instead of high-dose inorganic zinc sulfate can obviously improve the daily gain and feed conversion rate of the broiler chickens, and simultaneously has the important effects of improving the oxidation resistance and antibody level of organisms and improving the meat quality.

Application example 5: the application of argininyl zinc as feed additive for Penaeus vannamei Boone is provided.

The argininol zinc prepared in example 3 was added to a penaeus vannamei feed.

The test adopts a single-factor random grouping design, 800 tails of healthy young penaeus vannamei boone with the initial weight of about 0.350g are selected and randomly divided into 5 groups, each group has 4 repetitions, and each repetition has 40 tails. Wherein zinc sulfate (100ppm in terms of zinc) is added into basic daily ration in 1 group, and zinc sulfate is added into basic daily ration in 2 groups, 3 groups, 4 groups and 5 groups respectivelyArginine zinc (20ppm, calculated as zinc), arginine zinc (60ppm, calculated as zinc), arginine zinc (90ppm, calculated as zinc) and arginine zinc (120ppm, calculated as zinc) were added to the ration. The basic daily ration of each group takes fish meal and wheat protein powder as main protein sources. Test prawn, breeding in a volume of 0.38m³In the fiberglass steel drum. The cultivation process is carried out by referring to methods such as Yangqi Hui (2013) and the like. The test period was 8 weeks.

Detection indexes are as follows: (1) and (3) measuring growth performance indexes: the number and total weight of each shrimp replicate before the start were recorded. During the test, daily feeding and number of deaths, as well as body weight at death, were recorded. After the test was completed, the weight was repeated for each time after fasting for 24 hours. Survival (%) (100 x final shrimp mantissa/initial shrimp mantissa); the weight gain (%) was 100 × (final average weight-initial average weight)/initial average weight. (2) Blood index: weighing, collecting 10 shrimps in each case, collecting blood from heart, placing the blood sample in refrigerator at 4 deg.C overnight, centrifuging at 4 deg.C at 5000r/min for 10min, collecting serum, and storing at-80 deg.C. Serum lysozyme, phenoloxidase, and superoxide dismutase were performed according to the methods of LinShimei (2011) and the like.

TABLE 8 Effect of different treatments on Penaeus vannamei growth Performance and serum non-specific immune indices

As can be seen from Table 8 above, the weight gain of the 3, 4, 5 groups with 60ppm, 90ppm and 120ppm arginine zinc calculated as zinc was the highest, which was significantly higher than that of the 1 group with 100ppm zinc sulfate calculated as zinc (P <0.05), and there was no significant difference between the groups. And with the increase of the addition amount of the zinc argininate, the weight gain rate and the survival rate show an increasing trend. The 5-component activity of 120ppm arginino-zinc calculated as zinc was 2.92% higher (P > 0.05) than that of 1 group to which 100ppm zinc sulfate was added. The activities of lysozyme, phenol oxidase and SOD enzyme of 3, 4 and 5 groups of serum added with 60ppm, 90ppm and 120ppm arginine biotin zinc calculated by zinc are all obviously higher than those of 1 group and 2 group (P is less than 0.05). The activities of lysozyme, phenol oxidase and SOD enzyme of 5 groups of serum added with 120ppm argininyl zinc calculated by zinc are all significant for other groups (P is less than 0.05). And each blood index is increased along with the increase of the addition amount of the arginine biotin zinc. The result shows that the addition of 60-120 ppm arginine zinc calculated by zinc into the daily ration of the penaeus vannamei boone has obvious effects of enhancing the immunity of the penaeus vannamei boone, improving the survival rate and the growth performance compared with the addition of 100ppm zinc sulfate calculated by zinc.

The foregoing is considered as illustrative of the preferred embodiments of the invention and is not to be construed as limiting the invention in any way. Therefore, any simple modification, equivalent change and modification made to the above embodiments according to the technical spirit of the present invention should fall within the protection scope of the technical scheme of the present invention, unless the technical spirit of the present invention departs from the content of the technical scheme of the present invention.

Claims

1. A preparation method of arginine zinc capable of being used as an animal feed additive is characterized by comprising the following steps: reacting arginine biotin with carbonate or bicarbonate in water, stirring until no air bubbles emerge, heating, and adding inorganic zinc salt to react for a period of time to obtain arginine biotin zinc.

2. The method of claim 1, wherein the carbonate salt comprises sodium carbonate and/or potassium carbonate and the bicarbonate salt comprises sodium bicarbonate and/or potassium bicarbonate.

3. The preparation method according to claim 1 or 2, wherein the molar ratio of the carbonate to the argininyl is (1.01-1.02) to 1; the molar ratio of the bicarbonate to the argininol is (2.02-2.04) to 1.

4. The method of preparation according to claim 1 or 2, characterized in that the inorganic zinc salt comprises zinc chloride and/or zinc sulphate.

5. The preparation method according to claim 4, wherein the molar ratio of the inorganic zinc salt to the argininyl is (0.97-1.03) to 1.

6. The method according to claim 1 or 2, wherein the temperature is raised to not more than 60 ℃ and the period of time is 1.1 to 2 hours.

7. The production method according to claim 1 or 2, wherein the solid content of the reaction system is controlled to 18 to 45 wt%.

8. The preparation method according to claim 7, wherein when the solid content of the reaction system is 18-28%, after the reaction is completed, an organic solvent is added, and the mixture is filtered and dried, wherein the organic solvent is one or more of ethanol, propanol or isopropanol.

9. The application of the arginine biotin zinc prepared by the preparation method of any one of claims 1 to 8 as an animal feed additive, wherein the animal is a pig, a bird, a ruminant or an aquatic animal.

10. The use of claim 9, wherein the addition amount of the arginino-biotin zinc in the pig feed is 20-100 ppm calculated by zinc element; the addition amount of the zinc element in the poultry feed is 20-100 ppm; the addition amount of the zinc element in the aquatic feed is 20-120 ppm.