CN116898811A - Preparation method of rumen-protected nicotinic acid for preventing ketosis of dairy cows - Google Patents
Preparation method of rumen-protected nicotinic acid for preventing ketosis of dairy cows Download PDFInfo
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- CN116898811A CN116898811A CN202310748744.0A CN202310748744A CN116898811A CN 116898811 A CN116898811 A CN 116898811A CN 202310748744 A CN202310748744 A CN 202310748744A CN 116898811 A CN116898811 A CN 116898811A
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- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 title claims abstract description 109
- 235000001968 nicotinic acid Nutrition 0.000 title claims abstract description 64
- 239000011664 nicotinic acid Substances 0.000 title claims abstract description 64
- 229960003512 nicotinic acid Drugs 0.000 title claims abstract description 64
- 241000283690 Bos taurus Species 0.000 title claims abstract description 52
- 208000007976 Ketosis Diseases 0.000 title claims abstract description 26
- 230000004140 ketosis Effects 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 235000013365 dairy product Nutrition 0.000 title claims description 31
- 229920001661 Chitosan Polymers 0.000 claims abstract description 58
- 238000003756 stirring Methods 0.000 claims abstract description 54
- 210000004767 rumen Anatomy 0.000 claims abstract description 32
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 27
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 27
- 239000000661 sodium alginate Substances 0.000 claims abstract description 27
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 22
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 20
- 239000012153 distilled water Substances 0.000 claims abstract description 18
- 238000001694 spray drying Methods 0.000 claims abstract description 13
- 239000011162 core material Substances 0.000 claims abstract description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 55
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 51
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 24
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 claims description 13
- 239000012295 chemical reaction liquid Substances 0.000 claims description 12
- 239000000725 suspension Substances 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- 239000005457 ice water Substances 0.000 claims description 8
- 230000008569 process Effects 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 229940074391 gallic acid Drugs 0.000 claims description 6
- 235000004515 gallic acid Nutrition 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 4
- 239000008367 deionised water Substances 0.000 claims description 4
- 229910021641 deionized water Inorganic materials 0.000 claims description 4
- 239000012065 filter cake Substances 0.000 claims description 4
- 230000007935 neutral effect Effects 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 4
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 claims description 2
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 2
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims 1
- 230000002265 prevention Effects 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 22
- 238000012360 testing method Methods 0.000 description 10
- 230000000052 comparative effect Effects 0.000 description 9
- 230000000694 effects Effects 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 150000002576 ketones Chemical class 0.000 description 4
- 230000036542 oxidative stress Effects 0.000 description 4
- 230000009471 action Effects 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 230000006196 deacetylation Effects 0.000 description 3
- 238000003381 deacetylation reaction Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 230000031891 intestinal absorption Effects 0.000 description 2
- 230000006651 lactation Effects 0.000 description 2
- 238000000691 measurement method Methods 0.000 description 2
- 239000004005 microsphere Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 1
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000199919 Phaeophyceae Species 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 235000021050 feed intake Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- -1 lipid peroxides Chemical class 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/455—Nicotinic acids, e.g. niacin; Derivatives thereof, e.g. esters, amides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1611—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention discloses a preparation method of rumen-protected nicotinic acid for preventing ketosis of cows, belonging to the technical field of ketosis prevention of cows, comprising the following steps: adding sodium alginate into distilled water, and stirring the solution at 40-60 ℃ for 3-5h to obtain sodium alginate solution; adding the modified chitosan into absolute ethyl alcohol, and stirring for 20-30min to obtain chitosan solution; preparing a calcium chloride solution, mixing the calcium chloride solution with a chitosan solution, adding nicotinic acid, and stirring for 30-60min to obtain a core material solution; distilled water is added into the sodium alginate solution, a core material solution is dripped into the sodium alginate solution by using a syringe in a stirring state, after dripping is finished, the solution is kept stand, and spray drying is carried out while stirring, so that the rumen protected nicotinic acid is obtained.
Description
Technical Field
The invention belongs to the technical field of cow ketosis prevention, and particularly relates to a preparation method of rumen bypass protection nicotinic acid for preventing cow ketosis.
Background
Cows are in oxidative stress after delivery, and produce a large amount of oxygen free radicals and lipid peroxides, which cause oxidative damage to cells, reduced immune function and inflammatory response. At the same time, the body fat of the dairy cows is mobilized greatly, and excessive ketone bodies are accumulated in the bodies, so that ketosis of the dairy cows occurs.
Niacin, vitamin b5, is generally synthesized by microorganisms in the rumen of cows without the need for additional additions to the feed. However, the requirement of the dairy cows on nicotinic acid in the initial lactation period is higher than that of other dairy cows at any time, the physiological needs of the dairy cows can not be met by the nicotinic acid synthesized by the dairy cows, and if the dairy cows are not timely supplemented, the milk yield of the dairy cows is affected, and even the dairy cows are caused to generate ketosis.
The niacin can be largely degraded in the rumen of the dairy cows, the effective amount of the niacin is very low in absorption of the dairy cows, the palatability is poor, the ingestion of the dairy cows is easy to influence, so the niacin is not suitable for direct addition, the rumen bypass coating technology is adopted for protection, the degradation rate of the niacin in the rumen of the dairy cows is reduced, and the intestinal tract absorptivity is improved, but the effect of preventing ketosis of the dairy cows by the rumen bypass protection niacin is not ideal at present, firstly, the oxidative stress state of the dairy cows after delivery cannot be relieved by the existing rumen bypass protection niacin, secondly, more auxiliary materials and wall materials are adopted in the existing rumen bypass protection niacin preparation process, so that the niacin content in finished products is low, the absorption efficiency of animals during eating can be greatly reduced, and further the addition amount of the rumen bypass protection niacin in dairy cow feed is required to be improved, and the raising cost is increased.
Disclosure of Invention
The invention aims to provide a preparation method of rumen-protected nicotinic acid for preventing ketosis of cows, which aims to solve the problems that rumen-protected nicotinic acid cannot relieve oxidative stress state of cows after delivery and the content of nicotinic acid is low.
The aim of the invention can be achieved by the following technical scheme:
a preparation method of rumen protected nicotinic acid for preventing ketosis of cows comprises the following steps:
step S1, adding sodium alginate into distilled water, and stirring the solution for 3-5 hours at 40-60 ℃ to obtain sodium alginate solution;
s2, adding the modified chitosan into absolute ethyl alcohol, and stirring for 20-30min to obtain a chitosan solution;
step S3, preparing a calcium chloride solution, mixing the calcium chloride solution with the chitosan solution in the step S2, adding nicotinic acid, and stirring for 30-60min at the rotating speed of 100-300r/min to obtain a core material solution; and (3) adding distilled water into the sodium alginate solution in the step (S1), dropwise adding a core material solution at a speed of 2mL/min by using a syringe in a stirring state, standing for 25-30min after the dropwise adding, stirring while spray drying, and collecting a product at a discharge hole to obtain the rumen protected nicotinic acid.
Sodium alginate is a natural nontoxic polysaccharide found in brown algae, has the advantages of low cost, good biocompatibility, degradability and the like, has pH sensitivity, is easy to dissolve at high pH, is an enteric wall material, and chitosan is a biopolymer widely used as a carrier for gastrointestinal tract administration, so that the invention takes sodium alginate, modified chitosan, calcium chloride and nicotinic acid as raw materials, and the microsphere coated with the nicotinic acid is obtained by a microcapsule method, and then the product material is obtained by spray drying, and compared with the existing rumen protected nicotinic acid, the invention has high nicotinic acid content, can effectively inhibit the fat mobilization of dairy cows, improve the dry matter feed intake of the dairy cows and prevent ketosis of the dairy cows; in addition, the product can obviously reduce the release degree of nicotinic acid in gastric juice and improve the intestinal absorption effect; more prominently, the modified chitosan not only has the property of chitosan, forms a polymer capsule with sodium alginate, but also carries a gallic acid structure, can effectively remove oxygen free radicals, prevent lipid peroxidation, relieve the postpartum oxidative stress state of cows, and further prevent the cows from ketosis.
As a further technical scheme of the invention, the modified chitosan is prepared by the following steps:
placing purified chitosan into methanol, stirring to obtain suspension a, adding gallic acid into methanol, stirring, adding EDC, stirring for 0.5h, adding NHS, stirring under ice water bath for 1h to obtain reaction liquid b, adding reaction liquid b into suspension a, stirring under ice water bath for 0.5h, transferring to room temperature, stirring, reacting for 24h, centrifuging at 7500r/min for 10min, washing precipitate with methanol, and freeze drying to obtain modified chitosan.
As a further technical scheme of the invention, the dosage ratio of gallic acid, methanol, EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride), NHS (N-hydroxysuccinimide) and purified chitosan is 0.1g:10-20mL:0.12g:0.06-0.08g:0.3-0.5g, the purified chitosan and methanol in suspension a are used in a ratio of 1g:8-10 mL.
As a further technical scheme of the invention, the purified chitosan is prepared by the following steps:
dissolving chitosan in acetic acid solution, stirring for 8-12 hr, adding sodium hydroxide, standing for 6-8 hr, vacuum filtering, repeatedly washing filter cake with deionized water until the washing solution is neutral, and freeze drying to obtain chitosan with relative molecular weight of 7.2X10 5 The deacetylation degree (DD%) is 84%, and the dosage ratio of chitosan, acetic acid solution and sodium hydroxide is 1g:100mL:1g, acetic acid solution mass fraction is 0.5%.
As a further technical scheme of the invention, in the step S1, the mass ratio of sodium alginate to distilled water is 1.5-3.5:96.5-98.5.
As a further technical scheme of the invention, the dosage ratio of the modified chitosan to the absolute ethyl alcohol in the step S2 is 2.5-5g:250mL.
As a further technical scheme of the invention, in the step S3, the calcium chloride solution is calcium chloride aqueous solution with the mass fraction of 4-6%, and the dosage ratio of the calcium chloride solution, the chitosan solution, the nicotinic acid, the sodium alginate solution and the distilled water is 20-40mL:20-40mL:2-4g:10-20mL:70-140mL.
As a further technical scheme of the invention, the air inlet temperature is 160-190 ℃ in the spray drying process, the feeding flow rate is 5-20mL/min, and the air outlet temperature is 80-110 ℃.
The invention has the beneficial effects that:
the invention provides a preparation method for preventing rumen protected nicotinic acid of dairy cows, which comprises the steps of preparing enteric niacin microspheres by taking sodium alginate, modified chitosan and calcium chloride as coating materials, and spray drying to obtain the rumen protected nicotinic acid.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The embodiment provides a modified chitosan, which is prepared by the following steps:
30g of purified chitosan is placed in 240mL of methanol and stirred into suspension a, 10g of gallic acid is added into 1L of methanol, 12g of EDC is added after stirring, stirring is performed for 0.5h, 6g of NHS is added, stirring is performed under an ice water bath for 1h, reaction liquid b is obtained, the reaction liquid b is added into the suspension a, stirring is performed under the ice water bath for 0.5h, then the reaction liquid b is transferred to room temperature and stirred for 24h, finally the reaction liquid a is centrifuged for 10min at 7500r/min, and the precipitate is washed by methanol and freeze-dried, so that the modified chitosan is obtained.
Purified chitosan is prepared by the following steps:
dissolving 50g chitosan in 500mL 0.5wt% acetic acid solution, stirring for 8 hr, adding 50g sodium hydroxide, standing for 6 hr, suction filtering, repeatedly washing the filter cake with deionized water until the washing solution is neutral, and freeze drying to obtain chitosan with relative molecular weight of 7.2X10 5 The degree of deacetylation (DD%) was 84%.
Example 2
The embodiment provides a modified chitosan, which is prepared by the following steps:
50g of purified chitosan is placed in 500mL of methanol and stirred to form a suspension a, 10g of gallic acid is added into 2L of methanol, 12g of EDC is added after stirring, stirring is performed for 0.5h, 8g of NHS is added, stirring is performed under an ice water bath for 1h, reaction liquid b is obtained, the reaction liquid b is added into the suspension a, stirring is performed under the ice water bath for 0.5h, then the reaction liquid b is transferred to room temperature and stirred for 24h, finally the reaction liquid a is centrifuged for 10min at 7500r/min, and the precipitate is washed by methanol and freeze-dried, so that the modified chitosan is obtained.
Purified chitosan is prepared by the following steps:
dissolving 50g chitosan in 500mL 0.5wt% acetic acid solution, stirring for 12 hr, adding 50g sodium hydroxide, standing for 8 hr, suction filtering, repeatedly washing the filter cake with deionized water until the washing solution is neutral, and freeze drying to obtain chitosan with relative molecular weight of 7.2X10 5 The degree of deacetylation (DD%) was 84%.
Example 3
A preparation method of rumen protected nicotinic acid for preventing ketosis of cows comprises the following steps:
step S1, adding 1.5g of sodium alginate into 96.5g of distilled water, and stirring the solution for 3 hours at 40 ℃ to obtain sodium alginate solution;
step S2, adding 2.5g of the modified chitosan in the embodiment 1 into 250mL of absolute ethyl alcohol, and stirring for 20min to obtain a chitosan solution;
step S3, preparing a calcium chloride solution with the mass fraction of 4%, mixing 20mL of the calcium chloride solution with 20mL of the chitosan solution in the step S2, adding 2g of nicotinic acid, and stirring for 30min at the rotating speed of 100r/min to obtain a core material solution; adding 70mL of distilled water into 10mL of sodium alginate solution in the step S1, dropwise adding a core material solution at a speed of 2mL/min by using a syringe in a stirring state, standing for 25min after the dropwise adding, stirring while spray drying, and collecting a product at a discharge port to obtain the rumen protected nicotinic acid.
Wherein the air inlet temperature is 160 ℃ and the feeding flow rate is 5mL/min in the spray drying process, and the air outlet temperature is 80 ℃.
Example 4
A preparation method of rumen protected nicotinic acid for preventing ketosis of cows comprises the following steps:
step S1, adding 2.5g of sodium alginate into 97.5g of distilled water, and stirring the solution for 4 hours at 50 ℃ to obtain sodium alginate solution;
step S2, adding 4g of the modified chitosan in the embodiment 2 into 250mL of absolute ethyl alcohol, and stirring for 25min to obtain a chitosan solution;
step S3, preparing a calcium chloride solution with the mass fraction of 5%, mixing 30mL of the calcium chloride solution with 30mL of the chitosan solution in the step S2, adding 3g of nicotinic acid, and stirring at the rotating speed of 200r/min for 40min to obtain a core material solution; and (3) adding 110mL of distilled water into 15mL of sodium alginate solution in the step S1, dropwise adding a core material solution at a speed of 2mL/min by using a syringe in a stirring state, standing for 28min after the dropwise adding is finished, stirring while spray drying, and collecting a product at a discharge port to obtain the rumen protected nicotinic acid.
Wherein the air inlet temperature is 180 ℃ in the spray drying process, the feeding flow rate is 10mL/min, and the air outlet temperature is 100 ℃.
Example 5
A preparation method of rumen protected nicotinic acid for preventing ketosis of cows comprises the following steps:
step S1, adding 3.5g of sodium alginate into 98.5g of distilled water, and stirring the solution for 5 hours at 60 ℃ to obtain sodium alginate solution;
step S2, adding 5g of the modified chitosan in the embodiment 2 into 250mL of absolute ethyl alcohol, and stirring for 30min to obtain a chitosan solution;
step S3, preparing a calcium chloride solution with the mass fraction of 6%, mixing 40mL of the calcium chloride solution with 40mL of the chitosan solution in step S2, adding 4g of nicotinic acid, and stirring at the rotating speed of 300r/min for 60min to obtain a core material solution; and (3) adding 140mL of distilled water into 20mL of the sodium alginate solution in the step S1, dropwise adding a core material solution at a speed of 2mL/min by using a syringe in a stirring state, standing for 30min after the dropwise adding is finished, stirring while spray drying, and collecting a product at a discharge port to obtain the rumen protected nicotinic acid.
Wherein the air inlet temperature is 190 ℃ and the feeding flow rate is 20mL/min in the spray drying process, and the air outlet temperature is 110 ℃.
Comparative example 1
Compared with example 3, the modified chitosan in example 3 is replaced by purified chitosan in example 1, and the rest raw materials and the preparation process are the same as those in example 3.
The rumen protected nicotinic acid obtained in example 3-example 5 and comparative example 1 was tested as follows:
and (1) determination of niacin embedding rate: embedding rate (%) = (1-product surface niacin content/total niacin content in product) ×100%, product surface niacin content: weighing 0.2g of each group of products respectively, placing the products into a 10mL centrifuge tube, adding 8mL of methanol respectively, sufficiently shaking, filtering to obtain clear liquid, taking 0.1mL of clear liquid, fixing the volume to 1mL by using distilled water, taking 0.2mL of the solution at the moment, adding the solution into 10mL of specific color light, testing absorbance at 263nm wavelength according to an ultraviolet-visible spectrophotometry (Chinese pharmacopoeia 2010 edition two appendix IVA), and substituting the absorbance into a standard curve to obtain the content of surface niacin; total niacin content in the product: weighing 0.2g of each group of products, dissolving with distilled water, fixing the volume to 100mL, adding 0.2mL of sample liquid into a 10mL colorimetric tube, testing absorbance at 263nm wavelength according to an ultraviolet-visible spectrophotometry (Chinese pharmacopoeia 2010 edition II annex IV A), substituting the absorbance into a standard curve, and obtaining the total nicotinic acid content in the products, wherein the test result is shown in Table 1;
(II) gastrointestinal sustained release test: taking each group of samples, adopting a second method device of a dissolution rate measurement method (Chinese pharmacopoeia 2010 edition two-part annex XD) according to the dissolution rate measurement method (Chinese pharmacopoeia 2010 edition two-part annex XC), respectively taking 900mL of pH6.8 buffer solution and 900mL of pH1.2 buffer solution as medium solvents, carrying out normal operation at the rotation speed of 50 revolutions per minute, respectively taking 5mL of solution for filtration at 2, 4, 6 and 8 hours, timely supplementing 5mL of morphological solvent in an operation container, precisely measuring 1mL of continuous filtrate, placing the continuous filtrate into a 10mL measuring flask, diluting to a scale by phosphate buffer solution, testing absorbance at 263nm according to an ultraviolet-visible spectrophotometry (Chinese pharmacopoeia 2010 edition two-part annex IV A), respectively taking the release amounts of each group of products at different times in time according to the original developer U.S. Kos pharmaceutical company as reference preparations, and the test results are shown in tables 2 and 3;
(III) effect of preventing bovine ketosis: selecting a certain dairy farm in the Zhengzhen county of Anhui province, selecting 32 healthy Chinese Holstein cows with the child number of 2-3 and the lactation month of 4-5 months, randomly dividing into 4 groups according to a pairing principle, supplementing 1% rumen-passing protection nicotinic acid to each group of cows in basic ration, managing the test cows by special persons, feeding three times daily, feeding concentrate first, feeding coarse materials later, freely drinking water, mechanically milking three times daily, testing for 8 weeks, detecting blood ketone value (detected by using an abbott hemoketone meter) by taking blood of cow tails before the test starts (2022-10-31), taking an average value by each group, detecting blood ketone value by using the same method after the test, and observing the change of the blood ketone value of the cows, wherein the result is shown in table 4.
TABLE 1 niacin entrapment rate
| Project | Example 3 | Example 4 | Example 5 | Comparative example 1 |
| Embedding ratio (%) | 74.6 | 75.2 | 74.8 | 74.8 |
As can be seen from Table 1, the niacin embedding rates of the products obtained in examples 3-5 and comparative example 1 are both good.
Table 2 buffer Release at pH1.2
| Project | Example 3 | Example 4 | Example 5 | Comparative example 1 |
| 2h | 15.2 | 14.2 | 14.8 | 18.4 |
| 4h | 25.1 | 24.5 | 24.6 | 28.2 |
| 6h | 36.5 | 35.7 | 36.1 | 39.6 |
| 8h | 46.1 | 43.4 | 44.2 | 48.9 |
As can be seen from Table 2, the release rates of the products obtained in examples 3 to 5 and comparative example 1 under acidic conditions were small, indicating that the rumen bypass protection effect of the obtained products was good.
TABLE 3pH6.8 buffer
| Project | Example 3 | Example 4 | Example 5 | Comparative example 1 |
| 2h | 27.3 | 27.9 | 28.4 | 25.2 |
| 4h | 38.5 | 38.7 | 38.1 | 34.3 |
| 6h | 56.1 | 53.4 | 54.2 | 47.9 |
| 8h | 80.4 | 81.2 | 80.9 | 75.2 |
As can be seen from Table 3, the release rates of the products obtained in examples 3 to 5 and comparative example 1 were large under acidic conditions, indicating that the intestinal absorption effect of the obtained products was good.
TABLE 4pH6.8 buffer
As can be seen from Table 4, the products obtained in examples 3 to 5 have better effect of preventing bovine ketosis in cows than comparative example 1.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. The preparation method of the rumen protected nicotinic acid for preventing ketosis of dairy cows is characterized by comprising the following steps of:
step S1, adding sodium alginate into distilled water, and stirring the solution for 3-5 hours at 40-60 ℃ to obtain sodium alginate solution;
s2, adding the modified chitosan into absolute ethyl alcohol, and stirring for 20-30min to obtain a chitosan solution;
step S3, preparing a calcium chloride solution, mixing the calcium chloride solution with the chitosan solution in the step S2, adding nicotinic acid, and stirring for 30-60min to obtain a core material solution; and (3) adding distilled water into the sodium alginate solution in the step (S1), dropwise adding a core material solution by using a syringe in a stirring state, standing after the dropwise adding is finished, and performing spray drying while stirring to obtain the rumen protected nicotinic acid.
2. The preparation method of rumen protected nicotinic acid for preventing ketosis of dairy cows according to claim 1, wherein the modified chitosan is prepared by the following steps:
placing purified chitosan into methanol, stirring to obtain suspension a, adding gallic acid into methanol, stirring, adding EDC, stirring for 0.5h, adding NHS, stirring under ice water bath for 1h to obtain reaction liquid b, adding reaction liquid b into suspension a, stirring under ice water bath for 0.5h, transferring to room temperature, stirring, reacting for 24h, centrifuging at 7500r/min for 10min, washing precipitate with methanol, and freeze drying to obtain modified chitosan.
3. The method for preparing rumen protected nicotinic acid for preventing ketosis of dairy cows according to claim 2, wherein the dosage ratio of gallic acid, methanol, EDC, NHS and purified chitosan is 0.1g:10-20mL:0.12g:0.06-0.08g:0.3-0.5g, the purified chitosan and methanol in suspension a are used in a ratio of 1g:8-10 mL.
4. The method for preparing rumen protected nicotinic acid for preventing ketosis of dairy cows according to claim 1, wherein the purified chitosan is prepared by the following steps:
dissolving chitosan in acetic acid solution, stirring for 8-12h, adding sodium hydroxide, standing for 6-8h, suction filtering, repeatedly washing filter cake with deionized water until the washing solution is neutral, and freeze-drying to obtain the final product, wherein the dosage ratio of chitosan, acetic acid solution and sodium hydroxide is 1g:100mL:1g, acetic acid solution mass fraction is 0.5%.
5. The preparation method of rumen protected nicotinic acid for preventing ketosis of dairy cows according to claim 1, wherein the mass ratio of sodium alginate to distilled water in the step S1 is 1.5-3.5:96.5-98.5.
6. The preparation method of rumen bypass protection nicotinic acid for preventing ketosis of dairy cows according to claim 1, wherein the dosage ratio of modified chitosan to absolute ethyl alcohol in the step S2 is 2.5-5g:250mL.
7. The preparation method of rumen protected nicotinic acid for preventing ketosis of dairy cows according to claim 1, wherein in the step S3, the calcium chloride solution is a calcium chloride aqueous solution with the mass fraction of 4-6%, and the dosage ratio of the calcium chloride solution, the chitosan solution, the nicotinic acid, the sodium alginate solution and the distilled water is 20-40mL:20-40mL:2-4g:10-20mL:70-140mL.
8. The method for preparing rumen protected nicotinic acid for preventing ketosis of dairy cows according to claim 1, wherein the inlet air temperature is 160-190 ℃ in the spray drying process, the feeding flow rate is 5-20mL/min, and the outlet air temperature is 80-110 ℃.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110059162A1 (en) * | 2009-09-04 | 2011-03-10 | Jess Dreher Reed | Tannin-chitosan composites |
| CN107115315A (en) * | 2017-05-09 | 2017-09-01 | 北京东方天合生物技术有限责任公司 | Formula and preparation method for the rumen-protected nicotinic acid that prevents ketosis of dairy cows |
| CN110771742A (en) * | 2019-10-21 | 2020-02-11 | 安徽农业大学 | Feed additive for reducing methane emission of rumen of cattle and preparation method thereof |
| WO2022105301A1 (en) * | 2020-11-19 | 2022-05-27 | 广东海洋大学 | Asiaticoside-chitosan-sodium-alginate microsphere, and preparation method therefor and use thereof |
| US20220347341A1 (en) * | 2019-10-03 | 2022-11-03 | Medcura, Inc. | Hydrophobically-modified biopolymer materials |
| CN116195688A (en) * | 2023-03-16 | 2023-06-02 | 宁夏大学 | Functional feed additive for dairy cows, feed and preparation method |
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110059162A1 (en) * | 2009-09-04 | 2011-03-10 | Jess Dreher Reed | Tannin-chitosan composites |
| CN107115315A (en) * | 2017-05-09 | 2017-09-01 | 北京东方天合生物技术有限责任公司 | Formula and preparation method for the rumen-protected nicotinic acid that prevents ketosis of dairy cows |
| US20220347341A1 (en) * | 2019-10-03 | 2022-11-03 | Medcura, Inc. | Hydrophobically-modified biopolymer materials |
| CN110771742A (en) * | 2019-10-21 | 2020-02-11 | 安徽农业大学 | Feed additive for reducing methane emission of rumen of cattle and preparation method thereof |
| WO2022105301A1 (en) * | 2020-11-19 | 2022-05-27 | 广东海洋大学 | Asiaticoside-chitosan-sodium-alginate microsphere, and preparation method therefor and use thereof |
| CN116195688A (en) * | 2023-03-16 | 2023-06-02 | 宁夏大学 | Functional feed additive for dairy cows, feed and preparation method |
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