A branched-chain fatty acid preparation for improving calf growth performance and preparation method thereof

Abstract

The present invention belongs to the field of biomedicine and, in particular, relates to a branched-chain fatty acid preparation for improving calf growth performance and its preparation method. The branched-chain fatty acid preparation for improving calf growth performance comprises the following components by weight: 1-5 parts of a proanthocyanidin derivative, 3-10 parts of a mulberry leaf extract, 3-10 parts of a branched-chain fatty acid, and 0.3-0.8 parts of a trace element compound. The branched-chain fatty acid preparation for improving calf growth performance produced by the present invention can regulate rumen fermentation, reduce methane emissions and skatole content, thereby effectively reducing bloating symptoms and fecal odor in animals, providing a better growth environment for animals and promoting their growth and development.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a branched-chain fatty acid preparation for improving the growth performance of calves and a preparation method thereof. Background Art

[0002] At birth, the rumen of a calf is undeveloped and lacks a functional microbial community and ciliates. With the introduction of solid feeds, the rumen needs to adapt to this new nutritional regime through structural development (such as the formation of papillae) and functional improvement (such as improved volatile fatty acid metabolism). However, the rapid increase in starch and fiber in the diet during weaning often leads to fluctuations in ruminal pH and an imbalance in the microbial flora, which can cause bloat.

[0003] Branched-chain fatty acids (BCFAs) are a class of saturated fatty acids with a unique branched structure. Their carbon skeletons contain one or more branches (primarily methyl groups) and are classified as either single-branched or multi-branched. BCFAs, due to their branched structure, possess unique physical and chemical properties, such as a low freezing point, high thermal stability, and high oxidative stability. Their digestive and metabolic pathways in the body differ from those of straight-chain fatty acids, resulting in specialized physiological regulatory functions. The metabolic characteristics and functional properties of BCFAs may provide new insights into nutritional regulation during the specialized physiological phase of weaning in calves.

[0004] After weaning, calves' rumens develop rapidly, and the absorption of short-chain fatty acids becomes the primary energy source. However, during this period, forage intake increases, and as rumen microorganisms break down cellulose, they produce volatile fatty acids (such as acetic acid and propionic acid) and methane, which can easily lead to bloat. Furthermore, odor pollution generated during animal growth also seriously hinders the development of livestock and poultry farming. These odorous components are primarily generated and absorbed in the rumen, primarily due to unused nutrients in the animal's intestines and metabolites produced by microbial fermentation. In ruminant farming, reducing the content of odorous components is crucial for improving pasture environments and cattle growth and development.

[0005] Therefore, there is an urgent need to develop a branched-chain fatty acid preparation that can improve the growth performance of calves, regulate rumen fermentation, reduce methane emissions of calves, improve the growth environment, and promote the growth and development of calves. Summary of the Invention

[0006] In order to overcome the deficiencies of the prior art, one of the objectives of the present invention is to provide a branched-chain fatty acid preparation for improving the growth performance of calves.

[0007] One of the purposes of the present invention is achieved by the following technical solution:

[0008] The present invention provides a branched-chain fatty acid preparation for improving calf growth performance, comprising the following components in parts by weight: 1 to 5 parts of a proanthocyanidin derivative, 3 to 10 parts of a mulberry leaf extract, 3 to 10 parts of a branched-chain fatty acid, and 0.3 to 0.8 parts of a trace element compound; the proanthocyanidin derivative has the structure: .

[0009] Among them, the flavonoids (such as rutin and quercetin) and polyphenols in the mulberry leaf extract added in the present invention have broad-spectrum antibacterial activity, which can inhibit the excessive proliferation of gas-producing bacteria such as methanogens and hydrogen sulfide-producing bacteria in the rumen and reduce the production of methane.

[0010] Furthermore, the preparation process of the proanthocyanidin derivatives comprises the following steps:

[0011] (1) Ethyl (E)-4-hydroxy-2-butenoate, 7-chloroisoquinoline and potassium carbonate were added to N,N-dimethylformamide, heated for reaction, and purified to obtain intermediate 1;

[0012] (2) preparing an anhydrous tetrahydrofuran solution of intermediate 1, adding the anhydrous tetrahydrofuran solution of intermediate 1 to an anhydrous tetrahydrofuran solution of lithium aluminum tetrahydride to react, thereby obtaining intermediate 2;

[0013] (3) Add proanthocyanidin, butyric acid and triphenyl phosphate to anhydrous tetrahydrofuran, add diisopropyl azodicarboxylate, and stir to react to obtain a proanthocyanidin intermediate;

[0014] (4) The proanthocyanidin intermediate, intermediate 2, and silver oxide are added to a mixture of anhydrous toluene and anhydrous acetone in a volume ratio of 2:1 to react and obtain a proanthocyanidin derivative.

[0015] Furthermore, in step (1), the molar ratio of the (E)-4-hydroxy-2-butenoic acid ethyl ester, 7-chloroisoquinoline and potassium carbonate is 1: (1.2-1.5): (1.5-1.8); the concentration of the (E)-4-hydroxy-2-butenoic acid ethyl ester in N,N-dimethylformamide is 0.4 mol / L; the temperature of the heating reaction is 50-80° C., and the time is 10-16 h.

[0016] Furthermore, in step (2), the molar ratio of the intermediate 1 to lithium aluminum tetrahydride is 1:(1.5-2), the concentration of the anhydrous tetrahydrofuran solution of the intermediate 1 is 0.5 mol / L, and the concentration of the anhydrous tetrahydrofuran solution of lithium aluminum tetrahydride is 0.4 mol / L; when the anhydrous tetrahydrofuran solution of the intermediate 1 is added to the anhydrous tetrahydrofuran solution of lithium aluminum tetrahydride, the temperature of the reaction system is 0°C, the reaction temperature is room temperature, and the reaction time is 1-2 h.

[0017] Furthermore, in step (3), the molar ratio of the proanthocyanidin, butyric acid, triphenyl phosphate and diisopropyl azodicarboxylate is 1:(2-2.5):(2-2.5):(2-2.5), and the reaction time is 16-24 h; the amount ratio of the proanthocyanidin to anhydrous tetrahydrofuran is 1 mmol:8 mL.

[0018] Furthermore, the chemical structural formula of the proanthocyanidin is .

[0019] Furthermore, in step (4), the molar ratio of the proanthocyanidin intermediate, intermediate 2, and silver oxide is 1:(2.2-3):(2.8-3.5), the reaction temperature is 70-85°C, and the reaction time is 10-16 h; the amount ratio of the proanthocyanidin intermediate to the mixed solution is 1 mmol:7.5 mL.

[0020] Furthermore, the preparation process of the mulberry leaf extract is as follows: wash the mulberry leaves, dry them at 55°C for 5h, crush them and pass them through a 40-mesh sieve to obtain mulberry leaf powder, add the mulberry leaf powder to a 60v / v% ethanol solution, and the dosage ratio of mulberry leaf powder to ethanol solution is 1g:30mL. Ultrasonic extraction is performed at 50-60°C for 30-60min, centrifuged at 4000-5000rpm for 5-10min, the supernatant is taken, and lyophilized to obtain the mulberry leaf extract.

[0021] Furthermore, the branched fatty acid is any one of 12-methyltridecanoic acid, 13-methyltetradecanoic acid, and 14-methylpentadecanoic acid; and the trace element compound is any one of glycinate iron, sodium selenite, and glycinate zinc.

[0022] The second purpose of the invention is to provide a method for preparing a branched-chain fatty acid preparation for improving the growth performance of calves.

[0023] The second object of the present invention is achieved by adopting the following technical solution:

[0024] The present invention provides a method for preparing the above-mentioned branched-chain fatty acid preparation for improving calf growth performance, comprising the following steps: uniformly mixing a formulated amount of mulberry leaf extract, a proanthocyanidin derivative, a branched-chain fatty acid, and a trace element compound to obtain the branched-chain fatty acid preparation for improving calf growth performance.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] 1. The present invention provides a branched-chain fatty acid preparation for improving the growth performance of calves. Mulberry leaf extract, proanthocyanidin derivatives, branched-chain fatty acids, and trace elements are scientifically proportioned to prepare a branched-chain fatty acid preparation for improving the growth performance of calves. The branched-chain fatty acid preparation can regulate rumen fermentation, reduce methane emissions and skatole content, thereby effectively reducing the symptoms of bloating in animals, reducing fecal odor, and better promoting the growth and development of calves.

[0027] 2. This invention incorporates proanthocyanidin derivatives into the formulation. By incorporating a nitrogen-containing heterocyclic ring (similar in structure to the tryptophan group), the proanthocyanidin derivatives can compete with tryptophan for binding to tryptophan decarboxylase. This occupies the active site of tryptophan decarboxylase, preventing tryptophan from binding to the enzyme. This reduces the tryptophan metabolism rate and the conversion of tryptophan to indole metabolites (such as 3-methylindole), providing a favorable growth environment for calves. Furthermore, the butyrate group introduced into the proanthocyanidin structure promotes animal growth and improves intestinal health.

[0028] 3. The present invention provides a method for preparing the above-mentioned branched-chain fatty acid preparation for improving calf growth performance. The method is simple to operate and is conducive to industrial production. DETAILED DESCRIPTION

[0029] Below, in conjunction with specific embodiments, the present invention is further described. It should be noted that, under the premise of no conflict, the various embodiments described below or the various technical features can be arbitrarily combined to form new embodiments. Specific conditions not specified in the embodiments are carried out according to conventional conditions or conditions recommended by the manufacturer. Reagents or instruments used, unless otherwise specified, are conventional products obtained through commercial channels.

[0030] (1) Implementation

[0031] Example 1

[0032] This embodiment provides a branched-chain fatty acid preparation for improving calf growth performance, comprising the following components in parts by weight: 3 parts of a proanthocyanidin derivative, 7 parts of a mulberry leaf extract, 6 parts of 12-methyltridecanoic acid, and 0.5 parts of iron glycinate; the proanthocyanidin derivative has the structure: .

[0033] The preparation process of the proanthocyanidin derivatives includes the following steps:

[0034]

[0035]

[0036] (1) Ethyl (E)-4-hydroxy-2-butenoate (CAS: 10080-68-9), 7-chloroisoquinoline and potassium carbonate were added to N,N-dimethylformamide at a molar ratio of 1:1.4:1.6, wherein the concentration of ethyl (E)-4-hydroxy-2-butenoate in N,N-dimethylformamide was 0.4 mol / L. The mixture was heated to 65°C and reacted for 14 h. The reaction solution was cooled to room temperature, poured into water, and the aqueous phase was extracted with ethyl acetate. The ethyl acetate phase was washed with saturated brine and water in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate 1 (yield: 95%). 1 HNMR (C 15 H 15 NO3, DMSO-d6, 300 MHz): δ 9.19 (s, 1H), 8.43 (d, 1H), 7.58-7.50 (m, 3H), 7.12-7.07 (m, 2H), 6.06 (d, 1H), 4.67 (d, 2H), 4.06(q, 2H), 1.22 (t, 3H).ESI-MS(m / z): 258.11[M+H] + .

[0037] (2) Under nitrogen atmosphere, lithium aluminum tetrahydride was added to anhydrous tetrahydrofuran, stirred evenly to obtain a 0.4 mol / L lithium aluminum tetrahydride solution, which was cooled to 0°C. Intermediate 1 was then dissolved in anhydrous tetrahydrofuran to obtain a 0.5 mol / L solution of intermediate 1, which was then added dropwise to the lithium aluminum tetrahydride solution. The molar ratio of intermediate 1 to lithium aluminum tetrahydride was 1:1.7. After the addition was completed, the temperature was gradually raised to room temperature and the mixture was reacted at room temperature for 1.5 h. Water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate 2 (yield: 81.3%). 1 HNMR ( C 13 H 13 NO2, DMSO-d6, 300 MHz): δ 9.19 (s, 1H), 8.43 (d, 1H), 7.58-7.50 (m, 3H), 7.12(s, 1H), 5.89-5.87(m, 2H), 5.08(s, 1H), 4.69-4.67 (m,2H), 4.19-4.17(m, 2H). ESI-MS(m / z): 216.10[M+H] + .

[0038] (3) Proanthocyanidin, butyric acid, triphenyl phosphate and diisopropyl azodicarboxylate were weighed in a molar ratio of 1:2.2:2.2:2.2, and then, under a nitrogen atmosphere, proanthocyanidin, butyric acid and triphenyl phosphate (TPP) were added to anhydrous tetrahydrofuran in a ratio of 1 mmol:8 mL. Diisopropyl azodicarboxylate (DIAD) was then added at 0°C and stirred at room temperature for 20 h. The crude reaction mixture was purified by column chromatography to obtain (yield: 72%). Proanthocyanidin intermediate 1 HNMR ( C 38 H 38 O 15 , DMSO-d6, 300 MHz): δ 10.30 (s, 2H), 9.70 (s, 2H), 9.48 (s, 4H), 6.82-6.64 (m, 7H), 6.45 (d, 1H), 5.95-5.92 (m, 2H), 5.08(d,1H), 4.58 (q, 3H), 2.79-2.56 (m, 2H), 2.35 (t, 4H), 1.69-1.67 (m, 4H), 0.99(t, 6H). ESI-MS(m / z): 734.22[M].

[0039] (4) At room temperature, the proanthocyanidin intermediate, intermediate 2, and silver oxide were added in a molar ratio of 1:2.5:3 to a mixed solution of anhydrous toluene and anhydrous acetone (obtained by mixing anhydrous toluene and anhydrous acetone in a volume ratio of 2:1), wherein the amount ratio of the proanthocyanidin intermediate to the above mixed solution was 1 mmol:7.5 mL; stirred evenly at room temperature, heated to 80°C and reacted for 14 hours; cooled the reaction solution to room temperature and filtered, the filtrate was concentrated and purified by silica gel column chromatography (the volume ratio of dichloromethane to methanol was 15:1) to obtain a proanthocyanidin derivative (yield: 32.4%). Proanthocyanidin derivative 1 HNMR (C 64 H 60 N2O 19, DMSO-d6, 300 MHz): δ 10.30 (s, 2H), 9.70 (s, 2H), 9.19 (s, 2H), 8.43 (d, 2H), 7.58-7.50 (m, 6H), 7.13-7.10(m, 4H), 6.97 (d, 2H), 6.87 (d,2H), 6.70 (d, 1H), 6.45 (d, 1H), 5.95-5.88 (m, 4H), 5.08(d, 1H), 4.62-4.58(m, 3H), 4.42-4.20 (m, 4H), 4.10 (m, 2H), 4.00 (s, 2H), 3.80-3.74 (m, 4H), 2.79-2.56 (m, 2H), 2.35 (t, 4H), 1.69-1.67 (m, 4H), 0.99 (t, 6H). ESI-MS(m / z): 1160.40[M].

[0040] The preparation process of the above-mentioned mulberry leaf extract is as follows: fresh mulberry leaves are washed and dried, and then crushed through a 40-mesh sieve to obtain mulberry leaf powder, the mulberry leaf powder is added to a 60% volume fraction ethanol solution, wherein the amount ratio of mulberry leaf powder to ethanol solution is 1g:30mL, ultrasonic extraction is performed at 60°C for 60min, the extract is centrifuged at 4500rpm for 10min, the supernatant is concentrated and then freeze-dried to obtain the mulberry leaf extract.

[0041] This embodiment also provides a method for preparing the above-mentioned branched-chain fatty acid preparation for improving calf growth performance, comprising the following steps: charging a formulated amount of mulberry leaf extract, a proanthocyanidin derivative, 12-methyltridecanoic acid, and glycine iron into a mixer, and mixing to obtain the branched-chain fatty acid preparation for improving calf growth performance.

[0042] Example 2

[0043] This embodiment provides a branched-chain fatty acid preparation for improving calf growth performance, comprising the following components in parts by weight: 1 part of a proanthocyanidin derivative, 3 parts of a mulberry leaf extract, 3 parts of 13-methyltetradecanoic acid, and 0.3 parts of sodium selenite; the proanthocyanidin derivative has the structure: .

[0044] The preparation process of the proanthocyanidin derivatives includes the following steps:

[0045] (1) Ethyl (E)-4-hydroxy-2-butenoate, 7-chloroisoquinoline and potassium carbonate were added to N,N-dimethylformamide at a molar ratio of 1:1.2:1.5, wherein the concentration of ethyl (E)-4-hydroxy-2-butenoate in N,N-dimethylformamide was 0.4 mol / L. The mixture was heated to 50°C and reacted for 16 h. The reaction solution was cooled to room temperature, poured into water, and the aqueous phase was extracted with ethyl acetate. The ethyl acetate phase was washed with saturated brine and water in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate 1 (yield: 93.4%). 1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0046] (2) Under nitrogen atmosphere, lithium aluminum tetrahydride was added to anhydrous tetrahydrofuran, stirred evenly to obtain a 0.4 mol / L lithium aluminum tetrahydride solution, which was cooled to 0°C. Intermediate 1 was then dissolved in anhydrous tetrahydrofuran to obtain a 0.5 mol / L solution of intermediate 1, which was then added dropwise to the lithium aluminum tetrahydride solution. The molar ratio of intermediate 1 to lithium aluminum tetrahydride was 1:1.5. After the addition was completed, the temperature was gradually raised to room temperature and the mixture was reacted at room temperature for 1 h. Water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate 2 (yield: 80.1%). 1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0047] (3) Proanthocyanidin, butyric acid, triphenyl phosphate and diisopropyl azodicarboxylate were weighed in a molar ratio of 1:2:2:2, and then added to anhydrous tetrahydrofuran under a nitrogen atmosphere, wherein the amount ratio of proanthocyanidin to anhydrous tetrahydrofuran was 1 mmol:8 mL; then diisopropyl azodicarboxylate was added at 0°C and stirred at room temperature for 16 hours; the crude reaction mixture was purified by column chromatography to obtain (yield: 70.6%). Proanthocyanidin intermediate 1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0048] (4) At room temperature, the proanthocyanidin intermediate, intermediate 2, and silver oxide were added in a molar ratio of 1:2.2:2.8 to a mixed solution of anhydrous toluene and anhydrous acetone (obtained by mixing anhydrous toluene and anhydrous acetone in a volume ratio of 2:1), wherein the amount ratio of the proanthocyanidin intermediate to the above mixed solution was 1 mmol:7.5 mL; stirred evenly at room temperature, heated to 70°C and reacted for 16 hours; cooled the reaction solution to room temperature and filtered, the filtrate was concentrated and purified by silica gel column chromatography (the volume ratio of dichloromethane to methanol was 15:1) to obtain a proanthocyanidin derivative (yield: 30.5%). Proanthocyanidin derivative1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0049] The preparation process of the mulberry leaf extract is the same as that in Example 1.

[0050] This embodiment also provides a method for preparing the above-mentioned branched-chain fatty acid preparation for improving calf growth performance, comprising the following steps: charging a formulated amount of mulberry leaf extract, a proanthocyanidin derivative, 13-methyltetradecanoic acid, and sodium selenite into a mixer, and mixing to obtain the branched-chain fatty acid preparation for improving calf growth performance.

[0051] Example 3

[0052] This embodiment provides a branched-chain fatty acid preparation for improving calf growth performance, comprising the following components in parts by weight: 5 parts of a proanthocyanidin derivative, 10 parts of a mulberry leaf extract, 10 parts of 14-methylpentadecanoic acid, and 0.8 parts of zinc glycinate; the proanthocyanidin derivative has the structure: .

[0053] The preparation process of the proanthocyanidin derivatives includes the following steps:

[0054] (1) Ethyl (E)-4-hydroxy-2-butenoate, 7-chloroisoquinoline and potassium carbonate were added to N,N-dimethylformamide at a molar ratio of 1:1.5:1.8, wherein the concentration of ethyl (E)-4-hydroxy-2-butenoate in N,N-dimethylformamide was 0.4 mol / L. The mixture was heated to 80°C and reacted for 10 h. The reaction solution was cooled to room temperature, poured into water, and the aqueous phase was extracted with ethyl acetate. The ethyl acetate phase was washed with saturated brine and water in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate 1 (yield: 94.6%). 1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0055] (2) Under nitrogen atmosphere, lithium aluminum tetrahydride was added to anhydrous tetrahydrofuran, stirred evenly to obtain a 0.4 mol / L lithium aluminum tetrahydride solution, which was cooled to 0°C. Intermediate 1 was then dissolved in anhydrous tetrahydrofuran to obtain a 0.5 mol / L solution of intermediate 1, which was then added dropwise to the lithium aluminum tetrahydride solution. The molar ratio of intermediate 1 to lithium aluminum tetrahydride was 1:2. After the addition was completed, the temperature was gradually raised to room temperature and the mixture was reacted at room temperature for 2 h. Water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate 2 (yield: 81.2%). 1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0056] (3) Proanthocyanidin, butyric acid, triphenyl phosphate and diisopropyl azodicarboxylate were weighed in a molar ratio of 1:2.5:2.5:2.5, and then added to anhydrous tetrahydrofuran under a nitrogen atmosphere, wherein the amount ratio of proanthocyanidin to anhydrous tetrahydrofuran was 1 mmol:8 mL; diisopropyl azodicarboxylate (DIAD) was added at 0°C and stirred at room temperature for 24 hours; the crude reaction mixture was purified by column chromatography to obtain (yield: 71.3%). Proanthocyanidin intermediate 1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0057] (4) At room temperature, the proanthocyanidin intermediate, intermediate 2, and silver oxide were added to a mixed solution of anhydrous toluene and anhydrous acetone (obtained by mixing anhydrous toluene and anhydrous acetone in a volume ratio of 2:1) in a molar ratio of 1:3:3.5, wherein the amount ratio of the proanthocyanidin intermediate to the above mixed solution was 1 mmol:7.5 mL; stirred evenly at room temperature, heated to 85°C and reacted for 10 hours; cooled the reaction solution to room temperature and filtered, the filtrate was concentrated and purified by silica gel column chromatography (the volume ratio of dichloromethane to methanol was 15:1) to obtain a proanthocyanidin derivative (yield: 31.2%). Proanthocyanidin derivative 1 The results of HNMR and ESI-MS (m / z) were the same as those in Example 1.

[0058] The preparation process of the mulberry leaf extract is the same as that in Example 1.

[0059] This embodiment also provides a method for preparing the above-mentioned branched-chain fatty acid preparation for improving calf growth performance, comprising the following steps: charging a formulated amount of mulberry leaf extract, a proanthocyanidin derivative, 14-methylpentadecanoic acid, and zinc glycinate into a mixer, and mixing to obtain the branched-chain fatty acid preparation for improving calf growth performance.

[0060] (2) Comparative Example

[0061] Comparative Example 1

[0062] This comparative example 1 is basically the same as Example 1, except that the proanthocyanidin derivative is replaced by an equal amount of proanthocyanidin.

[0063] (3) Test examples

[0064] Test Example 1

[0065] Before the start of the feeding experiment, the feed troughs and water troughs in the cattle pen were thoroughly disinfected, and 3-month-old calves were selected as experimental animals. The calves were then randomly divided into 4 groups, with 7 calves in each group, namely: Example 1 Group, Example 2 Group, Example 3 Group, and Comparative Example 1 Group.

[0066] Group 1 of Example 1: basal diet and the straight-chain fatty acid preparation obtained in Example 1 accounting for 1% of the weight of the basal diet;

[0067] Group 2 of Example 2: basal diet and 1% of the basal diet weight of the straight-chain fatty acid preparation obtained in Example 2;

[0068] Example 3 group: basal diet and 1% of the basal diet by weight of the straight-chain fatty acid preparation obtained in Example 3;

[0069] Comparative Example 1 group: basal diet and the straight-chain fatty acid preparation obtained in Comparative Example 1 accounting for 1% of the weight of the basal diet;

[0070] The basal diet consisted of the following components by weight: 56.5% corn, 23.15% soybean meal, 10.5% bran, 6.5% corn distiller's grains with solubles, 2.3% rock powder, 0.55% calcium hydrogen phosphate, and 0.5% salt. Before each feeding, the basal diet was mixed evenly with the linear fatty acid preparation for the corresponding group. The diet was fed twice daily according to the above ratios, with ad libitum access to food and water for all groups. The pens were kept clean and hygienic.

[0071] (1) Before the start of the trial (Day 0) and after the end of the trial (Day 90), all cattle were weighed and their average daily weight gain (ADUG) was calculated using the formula: ADUG = (final weight - initial weight) / number of days in the trial. The results are recorded in Table 1.

[0072] Table 1

[0073]

[0074] In cattle fattening, daily weight gain is a key indicator of the fattening effect of Chinese Holstein calves. As shown in Table 1, the calves in Examples 1-3 weighed significantly more than those in Comparative Example 1 (where the proanthocyanidin derivatives were replaced with an equal amount of proanthocyanidins), demonstrating that the branched-chain fatty acid preparations prepared in this invention have advantages in promoting animal growth.

[0075] (2) Three days before the end of the experimental period, rumen fluid was collected through the mouth with a sterile oral catheter before morning feeding every day. After filtering with four layers of sterile gauze, the volatile fatty acid content (including acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid) of the collected rumen fluid was measured by gas chromatography. The average value of the three days was used as the final collection result. The experimental results are recorded in Table 2.

[0076] Table 2

[0077]

[0078] The rumen is a unique organ of ruminants, and its fermentation parameters are closely related to ruminant energy utilization. Rumen fluid volatile fatty acid production is often used to assess rumen fermentation. As shown in Table 2, the concentrations of volatile fatty acids in Examples 1-3 of the present invention were higher than those in Comparative Example 1. This result indicates active rumen fermentation, resulting in more efficient feed utilization, which in turn generates more energy for calf growth and contributes to increased calf weight.

[0079] (3) On the last day of the experiment (day 90), the methane emissions of calves in each group were measured using the American Sable open-circuit calorimetry system. The results are shown in Table 3.

[0080] Table 3

[0081]

[0082] As shown in Table 3, the methane emissions of Examples 1-3 of the present invention were as low as 52.36, significantly lower than those of Comparative Example 1. These results demonstrate that the addition of the linear fatty acid formulation of the present invention effectively reduces methane emissions. Reduced methane emissions mean more energy is available for cattle growth, improving feed conversion efficiency and promoting growth and development.

[0083] (4) On the last day of the experiment (day 90), the feces of each cow in the above groups were collected, and the content of skatole (3-methylindole) in the cow feces was determined by high performance liquid chromatography (HPLC). The test results are recorded in Table 4.

[0084] Table 4

[0085]

[0086] As shown in Table 4, the skatole content of Examples 1-3 of the present invention is much lower than that of Comparative Example 1. This result shows that the proanthocyanidin derivatives added to the branched fatty acid preparations of the present application play an important role. This is because the modified proanthocyanidin contains a nitrogen-containing heterocycle that is similar to the tryptophan group, which can compete with tryptophan for tryptophan decarboxylase, reduce the metabolic rate of tryptophan, and thus reduce the skatole content, providing a good growth environment for calves.

[0087] The above embodiments are only preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and replacements made by technicians in this field on the basis of the present invention fall within the scope of protection required by the present invention.

Claims

1. A branched-chain fatty acid preparation for improving calf growth performance, characterized in that: The invention comprises the following components in parts by weight: 1 to 5 parts of a proanthocyanidin derivative, 3 to 10 parts of a mulberry leaf extract, 3 to 10 parts of a branched-chain fatty acid, and 0.3 to 0.8 parts of a trace element compound; the chemical structural formula of the proanthocyanidin derivative is: ; The preparation process of the mulberry leaf extract is as follows: washing mulberry leaves, drying them at 55° C. for 5 hours, crushing them and passing them through a 40-mesh sieve to obtain mulberry leaf powder, adding the mulberry leaf powder to a 60 v / v% ethanol solution at a ratio of 1 g of mulberry leaf powder to 30 mL of ethanol solution, ultrasonically extracting the mulberry leaf at 50-60° C. for 30-60 minutes, centrifuging at 4000-5000 rpm for 5-10 minutes, collecting the supernatant, and freeze-drying the solution to obtain the mulberry leaf extract. The branched fatty acid is any one of 12-methyltridecanoic acid, 13-methyltetradecanoic acid, and 14-methylpentadecanoic acid; and the trace element compound is any one of glycinate iron, sodium selenite, and glycinate zinc.

2. The branched-chain fatty acid preparation for improving calf growth performance according to claim 1, characterized in that: The preparation process of the proanthocyanidin derivative comprises the following steps: (1) Ethyl (E)-4-hydroxy-2-butenoate, 7-chloroisoquinoline and potassium carbonate were added to N,N-dimethylformamide, heated for reaction, and purified to obtain intermediate 1; (2) preparing an anhydrous tetrahydrofuran solution of intermediate 1, adding the anhydrous tetrahydrofuran solution of intermediate 1 to an anhydrous tetrahydrofuran solution of lithium aluminum tetrahydride to react, thereby obtaining intermediate 2; (3) Add proanthocyanidin, butyric acid and triphenyl phosphate to anhydrous tetrahydrofuran, add diisopropyl azodicarboxylate, and stir to react to obtain a proanthocyanidin intermediate; (4) The proanthocyanidin intermediate, intermediate 2, and silver oxide are added to a mixture of anhydrous toluene and anhydrous acetone in a volume ratio of 2:1 to react and obtain a proanthocyanidin derivative.

3. The branched-chain fatty acid preparation for improving calf growth performance according to claim 2, characterized in that: In step (1), the molar ratio of the (E)-4-hydroxy-2-butenoic acid ethyl ester, 7-chloroisoquinoline and potassium carbonate is 1: (1.2-1.5): (1.5-1.8); the concentration of the (E)-4-hydroxy-2-butenoic acid ethyl ester in N,N-dimethylformamide is 0.4 mol / L; the temperature of the heating reaction is 50-80° C., and the time is 10-16 h.

4. The branched-chain fatty acid preparation for improving calf growth performance according to claim 2, characterized in that: In step (2), the molar ratio of the intermediate 1 to lithium aluminum tetrahydride is 1:(1.5~2), the concentration of the anhydrous tetrahydrofuran solution of the intermediate 1 is 0.5 mol / L, and the concentration of the anhydrous tetrahydrofuran solution of lithium aluminum tetrahydride is 0.4 mol / L; when the anhydrous tetrahydrofuran solution of the intermediate 1 is added to the anhydrous tetrahydrofuran solution of lithium aluminum tetrahydride, the temperature of the reaction system is 0°C, the reaction temperature is room temperature, and the reaction time is 1~2 hours.

5. The branched-chain fatty acid preparation for improving calf growth performance according to claim 2, characterized in that: In step (3), the molar ratio of the proanthocyanidin, butyric acid, triphenyl phosphate and diisopropyl azodicarboxylate is 1:(2-2.5):(2-2.5):(2-2.5), and the reaction time is 16-24 h; the amount ratio of the proanthocyanidin to anhydrous tetrahydrofuran is 1 mmol:8 mL.

6. The branched-chain fatty acid preparation for improving calf growth performance according to claim 2, characterized in that: In step (4), the molar ratio of the proanthocyanidin intermediate, intermediate 2, and silver oxide is 1:(2.2-3):(2.8-3.5), the reaction temperature is 70-85°C, and the reaction time is 10-16 h; the amount ratio of the proanthocyanidin intermediate to the mixed solution is 1 mmol:7.5 mL.

7. The method for preparing the branched-chain fatty acid preparation for improving calf growth performance according to any one of claims 1 to 6, characterized in that: The method comprises the following steps: uniformly mixing a formula amount of mulberry leaf extract, proanthocyanidin derivatives, branched-chain fatty acids and trace element compounds to obtain a branched-chain fatty acid preparation for improving calf growth performance.

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