25-hydroxyvitamin D3 premix as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of feed additives, and particularly relates to a 25-hydroxyvitamin D3 premix as well as a preparation method and application thereof, and the 25-hydroxyvitamin D3 premix comprises the following components in percentage by weight: 0.008%-0.08% of 25-hydroxyvitamin D3, 0.08%-0.7% of an emulsifier, 0.3%-1.2% of a stabilizer, 0.8%-1.7% of a synergist and the balance of a diluent. According to the 25-hydroxyvitamin D3 premixing agent, non-water-soluble raw materials are emulsified and coated into water-soluble raw materials, and the 25-hydroxyvitamin D3 premixing agent has the advantages of being simple in process, uniform in water dispersion, better in water solubility and stability and the like. The additive has the effects of improving the bone density of poultry, reducing the occurrence rate of special-shaped eggs in the later egg laying period of laying hens and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of feed additives, and specifically relates to a 25-hydroxyvitamin D3 premix, a preparation method and an application thereof. Background Art

[0002] At present, my country is the world's largest egg-laying chicken breeding and consumption country. As egg-laying chickens age, vitamin D3 metabolism becomes disordered, which will affect calcium absorption and deposition. Long-term egg-laying will also affect the thickness and strength of the eggshell. Calcium is the main component of the eggshell, accounting for about 38%-40% of the eggshell weight. Calcium deficiency in laying hens can easily lead to reduced egg production and thinner eggshells.

[0003] Vitamin D3 is involved in the entire process of calcium-phosphorus metabolism in the body. It is necessary for synthesizing calcium-binding proteins, activating bone calcium metabolism, and enhancing intestinal absorption of phosphorus metabolism. It is beneficial to adjust the calcium-phosphorus balance in the body.

[0004] 25-hydroxyvitamin D3 can directly enter the liver and then become active 1,25-2-hydroxyvitamin D3 in the kidneys, promoting calcium absorption and deposition in the eggshell. The biological efficacy of 25-hydroxyvitamin D3 is 3-5 times that of ordinary vitamin D3.

[0005] At present, most of the commercially available 25-hydroxyvitamin D3 products are liquid or water-insoluble powder products. Liquid products are easy to leak and unstable during transportation and storage. Insoluble products will bring troubles to farmers in the process of on-site use. Secondly, the addition amount is low and it is difficult to disperse evenly when mixed with feed. In addition, 25-hydroxyvitamin D3 is very sensitive to light and heat, and is easily oxidized and decomposed in the air, which will seriously affect the use of 25-hydroxyvitamin D3 products.

[0006] Therefore, it is very necessary to develop a stable water-soluble 25-hydroxyvitamin D3 premix product as a feed additive to improve eggshell quality in the later stage of egg laying and cope with high-temperature transportation and storage. Summary of the invention

[0007] The present invention aims to solve the technical problems in the prior art and provides a 25-hydroxyvitamin D3 premix and a preparation method and application thereof.

[0008] To achieve the above purpose, the specific technical solution of the present invention is as follows:

[0009] In one aspect, the present invention provides a 25-hydroxyvitamin D3 premix, which comprises 25-hydroxyvitamin D3, an emulsifier, a stabilizer, a synergist, and a diluent.

[0010] The components and mass percentages in the premix are: 25-hydroxyvitamin D3 0.008%-0.08%, emulsifier 0.08%-0.7%, stabilizer 0.3%-1.2%, synergist 0.8%-1.7%, and the diluent is supplemented to 100%.

[0011] Preferably, the components and mass percentages in the premix are: 25-hydroxyvitamin D3 0.01%-0.06%, emulsifier 0.1%-0.5%, stabilizer 0.5%-1%, synergist 0.9%-1.3%, and the diluent is supplemented to 100%.

[0012] More preferably, the components and mass percentages in the premix are: 25-hydroxyvitamin D3 0.03%, emulsifier 0.2%, stabilizer 0.8%, synergist 1.0%, and the diluent is supplemented to 100%.

[0013] The emulsifier is one or more of olive oil, Tween 80, tea oil, almond oil, glycerol, propylene glycol, and glyceryl monolaurate, preferably one or a combination of olive oil and Tween 80, and more preferably olive oil.

[0014] The stabilizer is one or more of tea polyphenols, vitamin E, tert-butylhydroquinone, ascorbic acid, glutathione, and taurine, preferably one or a combination of vitamin E and tert-butylhydroquinone, and more preferably vitamin E.

[0015] The synergist is one or more of vitamin D3, vitamin A, vitamin B2, crude extract of eucommia leaves, astragalus polysaccharide, and lipoic acid, preferably one or a combination of vitamin A, vitamin B2, crude extract of eucommia leaves, and lipoic acid.

[0016] The diluent is one or more of sodium sulfate, sodium bicarbonate, lactose, maltodextrin, and anhydrous glucose, preferably one or a combination of sodium bicarbonate and maltodextrin.

[0017] In another aspect, the present invention provides a method for preparing a 25-hydroxyvitamin D3 premix, comprising the following steps:

[0018] (1) Weigh 25-hydroxyvitamin D3, emulsifier, stabilizer, synergist, and diluent according to the mass fractions and set aside;

[0019] (2) heating the stabilizer and emulsifier to 50-90° C. to dissolve, adding the 25-hydroxyvitamin D3 and stirring to form an oil phase;

[0020] (3) putting the reaction solution obtained in step (2) into a homogenizer, adding a diluent and stirring to mix it evenly until it is in a moist state, then transferring it to a fluidized bed, blowing an air flow of 40° C. to 60° C., and continuing to spray the diluent to dry it to obtain a powder for use, and the homogenization pressure is not less than 30 MPa;

[0021] (4) The powder obtained in step (3) is passed through a 24-mesh sieve and mixed evenly with the synergist and the remaining diluent to obtain the product.

[0022] In another aspect, the present invention provides a 25-hydroxyvitamin D3 premix for use in improving the egg shell breakage rate of poultry in the late stage, improving the egg shell thickness, reducing the soft shell egg rate, and reducing the incidence of deformed eggs in laying hens in the late stage of egg production.

[0023] Preferably, the heating temperature in step (2) is 70°C.

[0024] The use method of the premix of the present invention is as follows: adding the premix to feed, 500 g of the product is added per ton of feed; adding the premix to drinking water, 250 g of the product is added per ton of water. Long-term addition of the premix has a better effect.

[0025] Beneficial effects:

[0026] 1. The present invention prepares 25-hydroxyvitamin D3 into a water-soluble premix after emulsification and coating, forming an oil-in-water state in water, which not only has higher stability but also has higher dispersibility, and is convenient for transportation and group administration.

[0027] 2. The premix obtained in the present invention can cope with high temperature and light conditions in the air, has higher stability under extreme conditions, reduces the impact of high temperature environments during production, storage, and transportation, and achieves a degradation rate far lower than that of commercially available products.

[0028] 3. The present invention has a significant effect on improving the egg shell breakage rate, egg shell thickness, and soft-shell eggs of laying hens in the later stage. DETAILED DESCRIPTION

[0029] The above contents will be further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter being limited to the following embodiments. All technologies realized based on the contents of the present invention are within the scope.

[0030] Example 1

[0031] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 802 g of Tween, 8 g of vitamin E, 2 g of eucommia leaf crude extract, 3 g of lipoic acid, 4 g of vitamin A, 1 g of vitamin B2, 600 g of sodium bicarbonate and 379.7 g of maltodextrin per 1 kg of the premix.

[0032] Example 2

[0033] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 2 g of olive oil, 8 g of tert-butylhydroquinone, 2 g of eucommia leaf crude extract, 3 g of lipoic acid, 4 g of vitamin A, 1 g of vitamin B2, 600 g of sodium bicarbonate and 379.7 g of maltodextrin in each 1 kg of the premix.

[0034] Example 3

[0035] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 2 g of olive oil, 8 g of vitamin E, 2 g of eucommia leaf crude extract, 3 g of lipoic acid, 4 g of vitamin A, 1 g of vitamin B2, 600 g of sodium bicarbonate and 379.7 g of maltodextrin per 1 kg of the premix.

[0036] Example 4

[0037] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 2 g of olive oil, 8 g of vitamin E, 2 g of eucommia leaf crude extract, 1 g of vitamin B2, 4 g of vitamin A, 600 g of sodium bicarbonate and 379.7 g of maltodextrin per 1 kg of the premix.

[0038] Example 5

[0039] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 1 g of olive oil, 1 g of Tween 80, 8 g of vitamin E, 4 g of lipoic acid, 1 g of vitamin B2, 4 g of vitamin A, 600 g of sodium bicarbonate and 379.7 g of maltodextrin per 1 kg of the premix.

[0040] Example 6

[0041] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 2 g of olive oil, 8 g of vitamin E, 2 g of eucommia leaf crude extract, 3 g of lipoic acid, 600 g of sodium bicarbonate and 379.7 g of maltodextrin per 1 kg of the premix.

[0042] Example 7

[0043] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 2 g of olive oil, 8 g of vitamin E, 2 g of astragalus polysaccharide, 3 g of lipoic acid, 4 g of vitamin A, 1 g of vitamin B2, 600 g of sodium bicarbonate and 379.7 g of maltodextrin per 1 kg of the premix.

[0044] Example 8

[0045] A 25-hydroxyvitamin D3 premix contains 0.3 g of 25-hydroxyvitamin D3, 2 g of olive oil, 8 g of ascorbic acid, 2 g of eucommia leaf crude extract, 3 g of lipoic acid, 4 g of vitamin A, 1 g of vitamin B2, 600 g of sodium bicarbonate and 379.7 g of maltodextrin in every 1 kg of the premix.

[0046] Example 9

[0047] A 25-hydroxyvitamin D3 premix contains 0.1 g of 25-hydroxyvitamin D3, 1 g of olive oil, 5 g of vitamin E, 2 g of eucommia leaf crude extract, 2 g of lipoic acid, 4 g of vitamin A, 1 g of vitamin B2, 600 g of sodium bicarbonate and 384.9 g of maltodextrin per 1 kg of the premix.

[0048] Example 10

[0049] A 25-hydroxyvitamin D3 premix contains 0.6 g of 25-hydroxyvitamin D3, 5 g of olive oil, 10 g of vitamin E, 3 g of eucommia leaf crude extract, 5 g of lipoic acid, 4 g of vitamin A, 1 g of vitamin B2, 600 g of sodium bicarbonate and 371.4 g of maltodextrin per 1 kg of the premix.

[0050] The method for preparing the 25-hydroxyvitamin D3 premix described in Examples 1-10 above comprises the following steps:

[0051] (1) Weigh 25-hydroxyvitamin D3, emulsifier, stabilizer, synergist, and diluent according to the mass fractions and set aside;

[0052] (2) After the stabilizer and emulsifier are heated to 70° C. to dissolve, the 25-hydroxyvitamin D3 is added and stirred to form an oil phase;

[0053] (3) putting the reaction solution obtained in step (2) into a homogenizer, adding a diluent and stirring to mix it evenly until it is in a moist state, then transferring it to a fluidized bed, blowing an air flow of 40° C. to 60° C., and continuing to spray the diluent to dry it to obtain a powder for use, and the homogenization pressure is not less than 30 MPa;

[0054] (4) The powder obtained in step (3) is passed through a 24-mesh sieve and mixed evenly with the synergist and the remaining diluent to obtain the product.

[0055] Comparative Example 1

[0056] An emulsion composition containing 25-hydroxyvitamin D3 was prepared by referring to the formulation and preparation process of Example 1 of Patent CN 107296152 A.

[0057] Test Example 1 25-Hydroxyvitamin D3 Water Solubility Dispersion Test

[0058] Experimental methods:

[0059] 1000 g of the products of Examples 1, 3, 5, 9, 10 and Comparative Example 1 were dissolved in 20 L of tap water at room temperature. After dissolution, 5 ml of the aqueous solution was taken from the upper, middle and lower points of the solution at 0, 8 and 24 hours, respectively. The solution was ultrasonicated for 30 min, the volume was made up to 50 ml with methanol, filtered with a 0.22 μm organic filter membrane, and the content was detected by liquid phase.

[0060] Liquid chromatography reference conditions:

[0061] aChromatographic column: C18 column, 4.6mm×250mm, particle size 5μm or equivalent.

[0062] b Mobile phase: Phase A is water, phase B is methanol, filtered through a 0.22 μm microporous organic filter membrane, and the gradient elution program is shown in Table 1.

[0063] cFlow rate: 1.0 mL / min.

[0064] d Column temperature: 40℃.

[0065] eInjection volume: 20μL.

[0066] f Wavelength: 265nm.

[0067] Table 1 Gradient elution program

[0068] Time, min Phase A, % Phase B, % 0.00 12 88 10.00 5 95 17.00 2 98 29.00 12 88 32.00 12 88

[0069] Determination:

[0070] Standard curve drawing

[0071] Accurately pipette the standard series solutions, inject the samples from low to high concentrations, and draw a standard curve with concentration as the horizontal axis and peak area as the vertical axis.

[0072] Quantitative determination:

[0073] Inject the test solution into the liquid chromatograph, substitute the result into the standard curve, and calculate the content by peak area according to the external standard method.

[0074] Calculation results:

[0075] The 25-hydroxyvitamin D3 content in the sample is calculated according to the following formula:

[0076]

[0077] Where:

[0078] W—25-hydroxyvitamin D3 in the sample, in milligrams per gram (mg / g);

[0079] c—the concentration of 25-hydroxyvitamin D3 in the sample solution obtained from the standard curve, in micrograms per milliliter, converted to milligrams per milliliter (mg / mL) according to the dilution multiple;

[0080] V—the final volume of the sample solution, in milliliters (mL);

[0081] m—sample mass, in grams (g);

[0082] The calculated result shall be rounded to three significant figures, and the absolute difference between two independent determination results obtained under repeatability conditions shall not exceed 10% of the arithmetic mean.

[0083] 1.2 Experimental results:

[0084] Table 2 Water-soluble dispersibility test

[0085]

[0086] As can be seen from the above table, the average decrease in 25-hydroxyvitamin D3 in Examples 3, 5, 9, and 10 of the present invention within 24 hours is relatively advantageous, all below 1%. In particular, the decreases in Example 3, Example 9, and Example 10 are the smallest, at 0.22%, 0.67%, and 0.22%, respectively. The maximum use time limit of the breeding site can be met. In addition, the difference in content between the upper and lower layers in Example 3 is 0.01 mg / kg, which is far better than 0.76 mg / kg in Comparative Example 1. It can be explained that after standing for 24 hours, the process Examples 3, 9-10 of the present invention can be evenly dispersed in water, meeting the requirements for water line use at each breeding farm.

[0087] Test Example 2 Comparison of the stability of various embodiments of the present invention

[0088] Experimental method: The products of Examples 1-3, 8-10 and Comparative Example 1 and commercially available 25-hydroxyvitamin D3 powder (with the same content as in Example 3) were placed in an oven at 60°C, and samples were taken on the 5th and 10th days to detect the content, thereby verifying the stability comparison between each example and Comparative Example 1.

[0089] Experimental Log:

[0090] Table 3 Stability test

[0091]

[0092]

[0093] From the above experimental results, it can be seen that the embodiment group of the present invention is relatively stable at 60°C for 5 days, without obvious degradation, and the degradation rate does not exceed 30% on the 10th day. The degradation rate of Example 3 is the lowest, which is 13.3%, followed by Example 9 and Example 10, which are 20% and 15%, respectively. Comparative Example 1 and the commercially available product both showed obvious degradation on the 5th day, and the degradation rate was particularly obvious on the 10th day. It can be shown that the process of the present invention can improve the effect of 25-hydroxy D3 at high temperature, which is more advantageous than commercially available products.

[0094] Test Example 3 Clinical Application Experiment

[0095] Experimental animals and groups:

[0096] In this experiment, a building of 540-day-old Hy-Line brown laying hens, about 40,500 chickens, were selected and randomly divided into 8 groups. Groups 1-7 were given the products of the present invention prepared in Examples 3-7, 9, and 10, respectively, mixed with drinking water, adding 50 g of the product of the present invention per ton of water, for 21 consecutive days; Group 8 was a blank control group, which was fed according to the normal feeding procedure, and the external environment of each group was the same.

[0097] Detection indicators:

[0098] Observe egg production rate and egg breakage rate

[0099] Experimental records and analysis:

[0100] Table 4 Improvement of eggshell by the additive of the present invention

[0101]

[0102]

[0103] As can be seen from the above table, the embodiments of the present invention have no obvious improvement effect on the egg production rate, but can maintain a stable egg production rate compared with the blank control group; however, there are relatively obvious effects on the egg breaking rate, among which the shell breaking rates of Example 3, Example 9, and Example 10 were reduced by 49.6%, 35.9%, and 47.1% respectively after 14 days of use compared with before use, while Examples 4-7 were reduced by 4.2%, 0.7%, 4.6%, and 16.4%, respectively, and the egg production rate and egg breaking rate of the blank control group were not improved.

[0104] Experimental Example 4: Clinical comparative test on the improvement effect of eggshell in laying hen

[0105] Experimental method: A 540-day-old Hailan brown laying hen was randomly divided into 6 groups. Groups 1-3 were the premixes prepared in Examples 3, 9, and 10, Group 4 was the commercially available product 1, and Group 5 was the commercially available product 2. The main ingredients of the experimental groups were all 25-hydroxyvitamin D3. Group 6 was the control group, which was given a normal diet. The experimental period was 30 days, and the dosage of each experimental group was the same. The mortality rate, soft-shelled egg rate, and feed-egg ratio were recorded before and after the experiment.

[0106] Table 5 Improvement of eggshell by the additive of the present invention

[0107] Group Mortality rate / % Soft-shelled egg rate / % Feed to Egg Ratio 1 group <![CDATA[1.95±0.5 ** ]]> <![CDATA[3.93±0.27 ** ]]> <![CDATA[2.31±1.46 * ]]> 2 groups <![CDATA[2.01±0.2 * ]]> <![CDATA[4.12±0.35 * ]]> 2.38±1.28 3 groups <![CDATA[1.96±0.6 ** ]]> <![CDATA[4.02±0.26 * ]]> 2.37±1.62 4 groups <![CDATA[2.01±0.4 * ]]> 4.44±0.47 2.39±1.33 5 groups 2.21±0.1 <![CDATA[4.35±0.36 * ]]> 2.39±1.56 6 groups 2.52±1.2 5.25±0.42 2.49±1.62

[0108] Note: * indicates significant difference between data in the same column (P<0.05), ** indicates extremely significant difference between data in the same column (P<0.01)

[0109] Experimental conclusion:

[0110] As can be seen from the above table, the first three experimental groups are more advantageous than the blank control group in terms of mortality rate, soft-shelled egg rate and feed-egg ratio, and the three indicators corresponding to the premixes prepared in Examples 3, 9 and 10 of the present invention (experimental groups 1-3) are significantly different from those of the other groups (P < 0.05), which shows that the product of the present invention is superior to the commercially available product group and can effectively reduce the mortality rate and feed-egg ratio.

[0111] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the technical principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A 25-hydroxyvitamin D3 premix, characterized in that: The components and mass percentages in the premix are: 25-hydroxyvitamin D3 0.008%-0.08%, emulsifier 0.08%-0.7%, stabilizer 0.3%-1.2%, synergist 0.8%-1.7%, and the diluent is supplemented to 100%.

2. The 25-hydroxyvitamin D3 premix according to claim 1, characterized in that The components and mass percentages in the premix are: 25-hydroxyvitamin D3 0.01%-0.06%, emulsifier 0.1%-0.5%, stabilizer 0.5%-1%, synergist 0.9%-1.3%, and the diluent is supplemented to 100%.

3. The 25-hydroxyvitamin D3 premix according to claim 1 or 2, characterized in that The components and mass percentages in the premix are: 25-hydroxyvitamin D3 0.03%, emulsifier 0.2%, stabilizer 0.8%, synergist 1.0%, and the diluent is supplemented to 100%.

4. The 25-hydroxyvitamin D3 premix according to any one of claims 1 to 3, characterized in that The emulsifier is one or more of olive oil, Tween 80, tea oil, almond oil, glycerol, propylene glycol, and monolaurin.

5. The 25-hydroxyvitamin D3 premix according to any one of claims 1 to 3, characterized in that: The stabilizer is one or more of tea polyphenols, vitamin E, tert-butylhydroquinone, ascorbic acid, glutathione and taurine.

6. The water-soluble 25-hydroxyvitamin D3 premix according to any one of claims 1 to 3, characterized in that: The synergist is one or more of vitamin D3, vitamin A, vitamin B2, crude extract of eucommia leaves, astragalus polysaccharide and lipoic acid.

7. The 25-hydroxyvitamin D3 premix according to any one of claims 1 to 3, characterized in that The diluent is one or more of sodium sulfate, sodium bicarbonate, lactose, maltodextrin and anhydrous glucose.

8. A method for preparing the 25-hydroxyvitamin D3 premix as claimed in any one of claims 1 to 3, characterized in that: The following steps are involved: (1) Weigh 25-hydroxyvitamin D3, emulsifier, stabilizer, synergist, and diluent according to the mass fractions and set aside; (2) heating the stabilizer and emulsifier to 50-90° C. to dissolve, adding the 25-hydroxyvitamin D3 and stirring to form an oil phase; (3) putting the reaction solution obtained in step (2) into a homogenizer, adding a diluent and stirring to mix it evenly until it is in a moist state, then transferring it to a fluidized bed, blowing an air flow of 40° C. to 60° C., and continuing to spray the diluent to dry it to obtain a powder for use, and the homogenization pressure is not less than 30 MPa; (4) The powder obtained in step (3) is passed through a 24-mesh sieve and mixed evenly with the synergist and the remaining diluent to obtain the product.

9. Use of the 25-hydroxyvitamin D3 premix as claimed in any one of claims 1 to 3 in improving the egg shell breakage rate of poultry in the late stage, improving the egg shell thickness, reducing the soft shell egg rate, and reducing the incidence of deformed eggs in laying hens in the late stage of egg production.

Images