A vitamin a derivative, its preparation and use

Abstract

The application belongs to the technical field of animal feed, and particularly relates to a vitamin A derivative, a preparation method and application thereof. The structural formula of the vitamin A derivative is shown as formula (I). The vitamin A derivative is added into daily feed, can replace the role of acetyl vitamin A and sodium butyrate, and has a favorable influence on the reproductive performance of SD rats in early pregnancy and the growth performance and intestinal health of weaned piglets.

Description

Technical Field

[0001] This invention belongs to the field of animal feed technology, specifically relating to a vitamin A derivative, its preparation method, and its application. Background Technology

[0002] Vitamin A is an essential nutrient. Vitamin A deficiency in pregnant SD rats can affect normal fetal development, while in piglets, it can lead to loss of appetite and stunted growth. Vitamin A plays a crucial regulatory role in maintaining intestinal homeostasis in piglets. Studies have shown that adding sodium butyrate to the diet can increase litter size in SD rats, reduce embryonic loss in early pregnancy, and have a beneficial effect on their reproductive performance. Currently, acetyl-retinyl acetate is commonly used in animal feed to supplement vitamin A.

[0003] In addition, sodium butyrate is now widely used in animal feed. It can promote the proliferation and maturation of gastrointestinal cells, maintain the beneficial microbial flora in the gastrointestinal tract, and improve animal production performance. Adding sodium butyrate to piglet diets can reduce post-weaning diarrhea, overcome weaning stress, and improve piglet survival rates. Summary of the Invention

[0004] This invention provides a vitamin A derivative, its preparation method, and its application. Adding this vitamin A derivative to the diet can replace the functions of acetyl vitamin A and sodium butyrate, and has a beneficial effect on the reproductive performance of SD rats in early pregnancy, as well as the growth performance and intestinal health of weaned piglets.

[0005] Specifically, the present invention provides the following technical solution:

[0006] A vitamin A derivative, with the structural formula shown in formula (I):

[0007]

[0008] The present invention also provides a method for preparing the above-mentioned vitamin A derivative, comprising the following steps:

[0009] 1) Dissolve retinol in an organic solvent;

[0010] 2) Add n-butyryl chloride and pyridine (acid-binding agent) to the solution obtained in step 1) and react for a certain time at a temperature of -4℃ to 2℃.

[0011] Preferably, the organic solvent is dichloromethane.

[0012] Preferably, the temperature of the solution is maintained between -4°C and 2°C during the addition of n-butyryl chloride and pyridine.

[0013] Preferably, the mass ratio of retinol, n-butyroyl chloride, and pyridine is 14-15:6.0-6.5:9.0-10.0.

[0014] Preferably, the reaction is carried out under stirring conditions.

[0015] Preferably, the specified time is 50 min to 70 min.

[0016] The present invention also provides the application of the above-mentioned vitamin A derivatives as alternatives to acetyl vitamin A and / or sodium butyrate in improving the reproductive performance of SD rats.

[0017] The present invention also provides the application of the above-mentioned vitamin A derivatives as alternatives to acetyl vitamin A and / or sodium butyrate in improving the growth performance of weaned piglets.

[0018] The present invention also provides the application of the above-mentioned vitamin A derivatives as alternatives to acetyl vitamin A and / or sodium butyrate in improving the intestinal health of weaned piglets.

[0019] This invention also provides a method for improving the reproductive performance of SD rats, comprising: feeding pregnant SD rats with the above-mentioned vitamin A derivative added to their diet. Preferably, 8000 IU to 12000 IU of vitamin A derivative is added per kilogram of diet.

[0020] This invention also provides a method for improving the growth performance of weaned piglets, comprising: adding the above-mentioned vitamin A derivative to the diet of weaned piglets. Preferably, 36,000 IU of vitamin A derivative is added per kilogram of diet.

[0021] The present invention also provides a method for improving the intestinal health of weaned piglets, comprising: feeding weaned piglets with the above-mentioned vitamin A derivative added to their diet.

[0022] The beneficial effects achieved by this invention are as follows:

[0023] This invention provides a vitamin A derivative, its preparation method, and its application. The preparation method is simple, low-cost, and has a high yield. Adding this vitamin A derivative to the diet can replace the functions of acetyl vitamin A and sodium butyrate, and has a beneficial effect on the reproductive performance of SD rats in early pregnancy, as well as the growth performance and intestinal health of weaned piglets. Attached Figure Description

[0024] Figure 1 This is the 1H NMR spectrum of a vitamin A derivative.

[0025] Figure 2 This represents the number of embryos implanted. Detailed Implementation

[0026] The following embodiments are used to illustrate the present invention, but are not intended to limit the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they should be performed in accordance with the techniques or conditions described in the literature in the field, or in accordance with the product manual.

[0027] In the following embodiments, instruments and other equipment whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels. Unless otherwise specified, the methods described are conventional methods, and the raw materials used are all available from publicly available commercial sources.

[0028] Method for synthesizing retinyl butyrate

[0029] 1. Synthesis of retinol

[0030] 1) Weigh 20g of retinyl acetate into a 1000mL single-necked flask, and add 150mL of ethanol and 150mL of isopropanol.

[0031] 2) Prepare a 10% KOH aqueous solution (10g KOH), add it to the above mixture, and stir. Heat at 90 degrees Celsius for 30 minutes.

[0032] 3) Ethanol and isopropanol were distilled off, and the residue was extracted three times with petroleum ether.

[0033] 4) Combine the extracts, dry them, remove the solvent, and dry the residue under vacuum using an oil pump to obtain retinol, which can be used directly in the next reaction.

[0034] 2. Synthesis of retinyl butyrate

[0035] 1) Dissolve 14-15g of the above retinol in 200mL of dichloromethane and cool to 0 degrees Celsius.

[0036] 2) Add 6.3 g of n-butyryl chloride and 9.5 g of pyridine, and stir at 0 degrees Celsius for 1 hour. The system is then rotary evaporated, and the residue is separated by column chromatography to obtain 9–10 g of retinyl butyrate. Figure 1 The 1H NMR spectrum is of retinyl butyrate (a vitamin A derivative).

[0037] Example 1

[0038] 1. Experimental Design and Diet

[0039] SD rats were placed in a constant-temperature environment and kept together overnight at a female-to-male ratio of 3:1. Vaginal smears were taken from the female rats at 8:00 AM the following morning to observe for pregnancy under a microscope, and the pregnant rats were weighed. The pregnant rats were randomly divided into five groups: a control group, a high-dose acetyl-vitamin A group, a high-dose butyryl-vitamin A group, a toxic-dose acetyl-vitamin A group, and a toxic-dose butyryl-vitamin A group. The control group was fed a basal pregnancy diet containing: digestible energy (3.48 kcal / g), crude protein (23.8%), crude fat (5.6%), crude fiber (4.4%), calcium (1.33%), and total phosphorus (0.96%). The HRA treatment group was fed a gestation diet containing 10,000 IU / kg of acetyl vitamin A; the HRB treatment group was fed a gestation diet containing 10,000 IU / kg of butyryl vitamin A; the PRA treatment group was fed a gestation diet containing 16,000 IU / kg of acetyl vitamin A; and the PRB treatment group was fed a gestation diet containing 1,600 IU / kg of butyryl vitamin A. 300g of each group's diet was placed in the feeding trough for each group. On day 7 of gestation, the rats were weighed at 8:00 AM after fasting. After anesthesia, the abdominal cavity of the rats was quickly opened, and the intact uterus was exposed using forceps. The number of embryos implanted in each rat was observed and recorded.

[0040] 2. Feeding and Management

[0041] This experiment was conducted in the Rat Nutrition and Metabolism Laboratory of the College of Animal Science and Technology, China Agricultural University (Beijing). The laboratory temperature was maintained at 22-26℃, with 12 hours of light per day and a relative humidity of approximately 50%. Three rats were housed in each experimental cage, with free access to food and water.

[0042] 3. Indicator Measurement and Methods

[0043] 3.1 Assessment of the number of embryos implanted in pregnant female mice

[0044] Fasting weights were taken at 8:00 a.m., and after anesthesia, the abdominal cavity of the rats was quickly opened. The intact uterus was exposed with forceps, and the number of embryos implanted in each rat was observed and recorded.

[0045] 4. Statistical Analysis

[0046] The experimental data were analyzed for variance using SPSS statistical software.

[0047] 5. Results

[0048] The mean number of embryo implantations was 14.69 in the control group, 16.23 in the treatment group (HRB), 14.47 in the treatment group (PRB), 14.5 in the treatment group (HRA), and 14.67 in the treatment group (PRA). Figure 2It was found that feeding SD rats a diet containing 10,000 IU / kg butyryl vitamin A significantly affected the number of embryo implantation in early pregnancy, significantly higher than other treatment groups. Therefore, adding a high dose of butyryl vitamin A to the diet increased the number of embryo implantation in early pregnancy in SD rats, which has a beneficial effect on the reproductive performance of SD rats.

[0049] Example 2

[0050] 1. Experimental Design and Diet

[0051] A total of 192 newly weaned piglets were selected for the experiment and divided into four groups, with six replicates per group and eight piglets per replicate. The experiment lasted for 28 days. On day 0, all piglets were weighed and randomly assigned to groups based on sex ratio and body weight. On days 14 and 28, piglets were weighed to calculate growth performance and feed intake. On days 14 and 28, one piglet from each replicate in each group was randomly selected for blood collection from the anterior vena cava. Diarrhea was recorded daily for each treatment, and a diarrhea record score was calculated. Fresh feces were collected from piglets on days 1, 7, 14, 21, and 28 for microbial testing. Since the vitamin A used in the piglet feed premix was acetylated vitamin A, a separate control group was not considered. This experiment designed two concentration gradients: at a concentration of 12,000 IU / kg, the feasibility of replacing acetylated vitamin A with butyryl vitamin A in a normal piglet diet was compared; at a high concentration of 36,000 IU / kg, the potential negative effects were compared. The specific composition of the normal basal feed is shown in Table 1, and the experimental groups are shown in Table 2.

[0052] Table 1. Dietary composition (%, feeding basal level)

[0053]

[0054]

[0055] The composition and proportions of the premix: Each kilogram of diet provides the following nutrients: Zinc, 60 mg; Iron, 95 mg; Copper, 10 mg; Iodine, 0.35 mg; Selenium, 0.3 mg; Manganese, 80 mg; Vitamin A, 12,000 IU; Vitamin D3, 2,750 IU; Vitamin E, 30 IU; Vitamin K3, 2 mg; Vitamin B12, 12 μg; Vitamin B2, 6 mg; Niacin, 40 mg; Pantothenic acid, 12 mg; Vitamin B6, 3 mg; Biotin, 0.2 mg.

[0056] Table 2 Experimental Groups

[0057]

[0058] 2. Feeding and Management

[0059] The experiment was conducted at the Hebei Fengning Animal Experiment Base of China Agricultural University. After weaning, piglets were fed in a nursery for 28 days, with 8 piglets per pen, fed twice a day (8:00 am and 3:00 pm), and had free access to water and food.

[0060] 3. Indicator Measurement and Methods

[0061] 3.1 Assessment of growth performance and severity of diarrhea

[0062] Individuals were weighed on an empty stomach in the morning at the start of the experiment, 14 days, and 28 days. Feed consumption was recorded in replicates per column, and average daily weight gain, average daily feed intake, and feed consumption-to-weight ratio were calculated.

[0063] The severity of diarrhea was monitored twice daily, and the consistency of feces was scored on a scale of 0 to 3 (0 = normal feces, 1 = soft stool, 2 = mild diarrhea, 3 = watery diarrhea). The scoring was performed by animal handlers who were unaware of the experimental treatment.

[0064] 4. Statistical Analysis

[0065] The experimental data were analyzed for variance using SPSS statistical software.

[0066] 5. Results

[0067] 5.1 Assessment of growth performance and diarrhea severity

[0068] The evaluation results are shown in Table 3.

[0069] Table 3. Effects of dietary supplementation with butyryl vitamin A on growth performance and diarrhea severity.

[0070]

[0071] In the table above, ADG represents average daily weight gain, ADFI represents average daily feed intake, FCR represents feed conversion ratio, SEM represents labeling error, and P value represents P value.

[0072] Diarrhea rate = Number of diarrhea episodes per pig / (Number of piglets × Number of days in the experiment);

[0073] Diarrhea index = (sum of piglet fecal scores throughout the entire experiment) / (number of piglets × number of days in the experiment).

[0074] As shown in the table, adding 36,000 IU / kg of butyryl vitamin A to the diet significantly improved the daily weight gain of weaned piglets compared to adding the same amount of acetyl vitamin A. Adding 36,000 IU / kg of butyryl vitamin A to the diet also significantly improved feed intake compared to adding 12,000 IU / kg of butyryl vitamin A. However, there were no significant differences in diarrhea rate and diarrhea index among the groups.

[0075] Therefore, adding different doses of acetyl vitamin A and butyryl vitamin A to the diet did not significantly affect the diarrhea in weaned piglets. However, compared with a diet containing 12,000 IU / kg of butyryl vitamin A, feeding piglets a diet containing 36,000 IU / kg of butyryl vitamin A significantly improved feed conversion ratio and daily weight gain. Therefore, feeding higher concentrations of butyryl vitamin A has a beneficial effect on the growth performance of weaned piglets.

[0076] Feeding different types of vitamin A has little effect on the growth performance of weaned piglets. Therefore, in actual production, butyryl vitamin A can be used as a substitute for acetyl vitamin A.

[0077] Example 3

[0078] 1. Experimental Design and Diet

[0079] The experiment selected 144 newly weaned piglets, divided into three groups, with 6 replicates per group and 8 piglets per replicate. The experiment lasted for 28 days. On day 0, all piglets were weighed and randomly assigned to groups based on sex ratio and body weight. On days 7, 14, and 21, piglets were weighed to calculate growth performance and feed intake. On days 7, 14, and 21, blood was collected from the anterior vena cava of one piglet from each replicate in each group. Diarrhea was recorded daily for each treatment, and a diarrhea record score was calculated. Fresh feces were collected on days 1, 3, 5, 7, 14, and 21 for microbiological testing. Treatment group A was fed a normal basal diet (containing 12000 IU / kg acetyl vitamin A), treatment group B was fed a normal basal diet supplemented with 24000 IU / kg acetyl vitamin A, and treatment group C was fed a normal basal diet supplemented with 24000 IU / kg butyryl vitamin A.

[0080] Example 3: The composition of the ordinary basal diet is the same as that in Example 2.

[0081] 2. Feeding and Management

[0082] The experiment was conducted at the Hebei Fengning Animal Experiment Base of China Agricultural University. After weaning, piglets were fed in a nursery for 21 days, with 8 piglets per pen, fed twice a day (8:00 am and 3:00 pm), and had free access to water and food.

[0083] 3. Indicator Measurement and Methods

[0084] 3.1 Assessment of growth performance and severity of diarrhea

[0085] Individuals were weighed on an empty stomach in the morning at the start of the experiment, 7 days, 14 days, and 21 days. Feed consumption was recorded in replicates per column, and average daily weight gain, average daily feed intake, and feed consumption-to-weight ratio were calculated.

[0086] The severity of diarrhea was monitored twice daily, and the consistency of feces was scored on a scale of 0 to 3 (0 = normal feces, 1 = soft stool, 2 = mild diarrhea, 3 = watery diarrhea). The scoring was performed by animal handlers who were unaware of the experimental treatment.

[0087] 4. Statistical Analysis

[0088] The experimental data were analyzed for variance using SPSS statistical software.

[0089] 5. Results

[0090] 5.1 Assessment of growth performance and diarrhea severity

[0091] The evaluation results are shown in Table 4.

[0092] Table 4. Effects of dietary supplementation with butyryl vitamin A on growth performance and diarrhea severity.

[0093]

[0094] As shown in the table, the addition of butyryl vitamin A or acetyl vitamin A 24000 IU / kg to the basal diet can significantly increase the feed intake and daily weight gain of weaned piglets, but there are no significant differences in diarrhea rate and diarrhea index among the groups.

[0095] Therefore, adding different doses of acetyl vitamin A and butyryl vitamin A to the diet did not significantly affect diarrhea in weaned piglets. However, compared with feeding piglets a regular diet (containing acetyl vitamin A 12000 IU / kg), diets supplemented with 24000 IU / kg of butyryl vitamin A or acetyl vitamin A 24000 IU / kg showed significantly increased feed intake and daily weight gain. Therefore, feeding higher concentrations of acetyl vitamin A or butyryl vitamin A has a beneficial effect on the growth performance of weaned piglets. At the same dosage, feeding different types of vitamin A had little effect on the growth performance of weaned piglets.

[0096] Example 4

[0097] 1. Experimental Design and Diet

[0098] Ninety-six newly weaned piglets were selected for the experiment and divided into three groups, with four replicates per group and eight piglets per replicate. The experiment lasted for 28 days. On day 0, all piglets were weighed and randomly assigned to groups based on sex ratio and body weight. On days 14 and 28, piglets were weighed to calculate growth performance and feed intake. On days 14 and 28, blood was collected from the anterior vena cava of one piglet from each replicate in each group. Diarrhea was recorded daily for each treatment, and a diarrhea record score was calculated. Fresh feces were collected on days 1, 3, 5, 7, 14, 21, and 28 for microbial testing. Treatment group A was fed a standard basal diet, treatment group B was fed a standard basal diet supplemented with 0.1% sodium butyrate, and treatment group C was fed a standard basal diet supplemented with 12000 IU / kg butyryl vitamin A.

[0099] Example 4: The composition of the ordinary basal diet is the same as that in Example 2.

[0100] 2. Feeding and Management

[0101] The experiment was conducted at the Hebei Fengning Animal Experiment Base of China Agricultural University. After weaning, piglets were fed in a nursery for 28 days, with 8 piglets per pen, fed twice a day (8:00 am and 3:00 pm), and had free access to water and food.

[0102] 3. Indicator Measurement and Methods

[0103] 3.1 Assessment of growth performance and severity of diarrhea

[0104] Individuals were weighed on an empty stomach in the morning at the start of the experiment, 14 days, and 28 days. Feed consumption was recorded in replicates per column, and average daily weight gain, average daily feed intake, and feed consumption-to-weight ratio were calculated.

[0105] The severity of diarrhea was monitored twice daily, and the consistency of feces was scored on a scale of 0 to 3 (0 = normal feces, 1 = soft stool, 2 = mild diarrhea, 3 = watery diarrhea). The scoring was performed by animal handlers who were unaware of the experimental treatment.

[0106] 4. Statistical Analysis

[0107] The experimental data were analyzed for variance using SPSS statistical software.

[0108] 5. Results

[0109] 5.1 Assessment of growth performance and diarrhea severity

[0110] The evaluation results are shown in Table 5.

[0111] Table 5. Effects of dietary supplementation with butyryl vitamin A on growth performance and diarrhea severity.

[0112]

[0113] As shown in the table, adding 12,000 IU / kg of butyryl vitamin A to the diet significantly improved the feed conversion ratio of weaned piglets compared with adding 0.1% sodium butyrate and the basal diet. However, there were no significant differences in diarrhea rate and diarrhea index among the groups.

[0114] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. Application of vitamin A derivatives as substitutes for acetyl-vitamin A and / or sodium butyrate in the preparation of diets to improve the reproductive performance of SD rats, wherein the structural formula of the vitamin A derivative is shown in formula (I): （I）。 2. Application of vitamin A derivatives as substitutes for acetyl vitamin A and / or sodium butyrate in the preparation of diets to improve the growth performance of weaned piglets, wherein the structural formula of the vitamin A derivative is shown in formula (I): （I）。 3. A method for preparing a diet to improve the reproductive performance of SD rats, characterized in that, include: Pregnant SD rats were fed a diet containing a vitamin A derivative; 10,000 IU of the vitamin A derivative was added per kilogram of diet, and the structural formula of the vitamin A derivative is shown in formula (I). （I）。 4. A method for preparing a diet to improve the growth performance of weaned piglets, characterized in that, include: Weaned piglets were fed a diet containing a vitamin A derivative; 36,000 IU of the vitamin A derivative was added per kilogram of diet, and the structural formula of the vitamin A derivative is shown in formula (I): （I）。

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