CN114698738B - Rumen bypass schizochytrium limacinum powder and preparation method thereof - Google Patents
Rumen bypass schizochytrium limacinum powder and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
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- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
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Abstract
The application relates to the field of animal feed, and in particular discloses rumen bypass schizochytrium limacinum powder and a preparation method thereof. The preparation method of the rumen bypass schizochytrium limacinum powder comprises the following steps: uniformly mixing the schizochytrium limacinum powder with deionized water, adding mesoporous silicon dioxide, carrying out ultrasonic treatment at room temperature, heating to 37-40 ℃, uniformly stirring, centrifuging, and drying to prepare mesoporous silicon dioxide loaded schizochytrium limacinum powder; uniformly mixing mesoporous silica-loaded schizochytrium limacinum powder with dichloromethane and ethanol, adding zein, uniformly mixing, introducing supercritical carbon dioxide, boosting to 8-10MPa, preserving heat at 45-55 ℃ for 90-100min, cooling, and drying to obtain a core material; heating palm oil fat powder to a molten state, dissolving corn fiber glue in water, mixing with the molten state palm oil fat powder, and uniformly mixing to prepare a wall material; and (5) coating. The rumen bypass schizochytrium limacinum powder has the advantages of high rumen bypass rate, high DHA effective utilization rate and good storage stability.
Description
Technical Field
The application relates to the field of animal feed, in particular to rumen bypass schizochytrium limacinum powder and a preparation method thereof.
Background
Docosahexaenoic acid, also known as DHA, plays an important role in human growth and development. DHA has effects of reducing blood lipid, promoting fetal brain cell growth, strengthening nervous system, improving vision, delaying brain aging, and maintaining cardiovascular and cerebrovascular system health. The human body cannot synthesize DHA by itself and needs to be supplemented by food absorption, such as food deep sea fish, nuts, DHA products and the like.
With the continuous improvement of health demands and the enhancement of health consciousness of people, the importance of DHA is generally known, the existing food cannot meet the demands of consumers, and the egg and the milk rich in DHA are effective means for supplementing DNA through diet. The schizochytrium limacinum powder is rich in DHA, can be used as a feed additive to be added into daily ration of livestock, and can improve the DHA content in livestock products. However, due to the special digestive structure of ruminants, most of the DHA supplied in unprotected form is acted by microorganisms in the rumen, hydrogenation occurs, resulting in a low DHA content that effectively enters and is absorbed by the intestinal tract, and eventually a small DHA content is accumulated in the milk.
In view of the above related art, the inventors found that the stability of schizochytrium limacinum powder in the rumen environment is insufficient and the utilization rate is to be improved.
Disclosure of Invention
In order to improve the stability of the schizochytrium limacinum powder in the rumen, increase the release rate of the schizochytrium limacinum powder after entering the small intestine and improve the effective utilization rate, the application provides the rumen bypass schizochytrium limacinum powder and a preparation method thereof.
In a first aspect, the present application provides a preparation method of rumen bypass schizochytrium limacinum powder, which adopts the following technical scheme: a preparation method of rumen bypass schizochytrium limacinum powder comprises the following steps:
preparing a core material: (1) Uniformly mixing the schizochytrium limacinum powder with deionized water, adding mesoporous silicon dioxide, carrying out ultrasonic treatment at room temperature for 30-40min, heating to 37-40 ℃, uniformly stirring, centrifuging, and drying to obtain mesoporous silicon dioxide loaded schizochytrium limacinum powder;
(2) Uniformly mixing mesoporous silica-loaded schizochytrium limacinum powder with dichloromethane and ethanol, adding zein, uniformly mixing, introducing supercritical carbon dioxide, boosting to 8-10MPa, preserving heat at 45-55 ℃ for 90-100min, cooling to room temperature, and drying to obtain a core material;
preparing a wall material: heating palm oil fat powder to a molten state, dissolving corn fiber glue in water, mixing with the molten state palm oil fat powder, and uniformly mixing to prepare a wall material;
coating: mixing wall material, core material and citric acid, regulating pH to 8-10, heating to 90-95deg.C, stirring, and coating to obtain the final product.
By adopting the technical scheme, the surface of the mesoporous silica has a porous communication structure, small pore diameter, low relative density, high specific strength, high porosity and specific surface area, good permeability and adsorptivity, large loading capacity, controllable release behavior, certain targeting property, self insolubility in water, gradual dissolution after degradation of gastrointestinal enzymes in vivo, so that the schizochytrium limacinum powder and the mesoporous silica are ultrasonically mixed, the schizochytrium limacinum powder is loaded on the mesoporous silica, then the mesoporous silica filled with the schizochytrium limacinum powder is coated in a zein film by adopting a supercritical carbon dioxide anti-solvent method, the mesoporous silica is insoluble in an organic solvent due to the fact that the zein is dissolved in dichloromethane and ethanol, and is dispersed in a solution in a form of particles to form a suspension, and the supercritical carbon dioxide is used as an anti-solvent to contact with the suspension, so that the situation that the mesoporous silica and the zein are separated out simultaneously or the mesoporous silica is separated out and embedded unsuccessfully is avoided; the mesoporous silica loaded with the schizochytrium limacinum powder is diffused from the zein, and is released continuously along with the damage to the wall structure of the zein, so that the stability of the zein in releasing the mesoporous silica loaded with the schizochytrium limacinum powder is improved, the sustained release effect is achieved, the digestion speed is slowed down, the digestion time of the schizochytrium limacinum powder in the intestinal tract is improved, and the utilization rate is prolonged.
The palm oil fat powder is used as a wall material, is not easy to degrade in rumen, and is digested and absorbed in small intestine, the corn fiber gum is a natural polysaccharide extracted from corn fiber, has a highly branched arabinoxylan structure, has adhesiveness, and can effectively improve the stability of the wall material and the connection stability of the wall material and a core material; the palm oil fatty acid powder and the corn fiber gum are used as wall materials, so that the degradation of the schizochytrium limacinum powder by microorganisms in the rumen can be reduced, DHA is protected to be released after entering the small intestine and is absorbed and utilized by the organism, the conversion rate of DHA in the organism of the animal is improved, the loss of nutrient substances is reduced, and the utilization rate of the feed is improved.
Preferably, the mass ratio of the core material to the wall material is 0.3-0.5:1;
in the core material, the mass ratio of the schizochytrium limacinum powder to the mesoporous silica to the zein is 1:4-5:7-10;
in the wall material, the mass ratio of the palm oil fat powder to the corn fiber gum is 1:0.6-1.
By adopting the technical scheme, the amount of the schizochytrium limacinum powder, the mesoporous silica and the zein in the core material is controlled by adopting the technical scheme, so that the schizochytrium limacinum powder can be fully filled in the mesoporous silica, the mesoporous silica is coated in the zein, and a certain amount of corn fiber glue is added in the palm oil fat powder, so that the structural stability of the wall material can be improved.
Preferably, in the step (1), the mass ratio of the schizochytrium limacinum powder to the deionized water is 1:3-5;
the mass ratio of dichloromethane to ethanol to zein in the step (2) is 9:5:7-10.
By adopting the technical scheme, the schizochytrium limacinum powder and water are fully mixed in the step (1), so that the schizochytrium limacinum powder is uniformly loaded in the mesoporous silica, and the loading capacity is increased; in the step (2), dichloromethane and ethanol are mixed, so that zein can be effectively dissolved, the coating rate of zein on mesoporous silica is high, and the coating is tight.
Preferably, the flow rate of the carbon dioxide is 2-3kg/h.
By adopting the technical scheme, the zein can be uniformly coated on the mesoporous silica, and the coating rate is improved.
Preferably, the mesoporous silica is pretreated by:
(1) Adding mesoporous silica into ammonium persulfate aqueous solution with mass concentration of 18-20%, stirring at room temperature for 20-24h, centrifuging, washing the centrifugate with distilled water until the pH value is neutral, and drying to obtain oxidized mesoporous silica;
(2) Adding the mesoporous silica into deionized water, carrying out ultrasonic treatment for 20-30min, adding dodecylamine and ethanol solution, heating to 80-90 ℃, stirring for 20-24h, and drying.
Through the technical scheme, mesoporous silica is mixed with ammonium persulfate, after the mesoporous silica is modified by an acidic ammonium persulfate solution, oxygen-containing groups are introduced into the surface of the mesoporous silica, poor dispersibility of the mesoporous silica in an aqueous solution is remarkably improved by oxidation modification, the structure of the mesoporous silica is not damaged by oxidation modification, but the hydrophilicity of the oxidized mesoporous silica is increased, and when the mesoporous silica is coated in zein by supercritical carbon dioxide fluid, the hydrophilicity of the mesoporous silica is improved, so that the solubility of the mesoporous silica in ethanol and dichloromethane is increased, and the load capacity is reduced; therefore, the oxidized mesoporous silica is mixed with the dodecylamine, nucleophilic substitution reaction is carried out between oxygen-containing functional groups on the oxidized mesoporous silica and the dodecylamine, the alkylamine with long carbon chains is grafted onto the oxidized mesoporous silica in a covalent bond mode, the hydrophobicity of the oxidized mesoporous silica modified by the dodecylamine is enhanced, so that the solubility of the oxidized mesoporous silica in ethanol is reduced, the encapsulation rate of the mesoporous silica loaded with the schizochytrium limacinum powder in the zein is improved, the release amount of the schizochytrium limacinum powder is increased, and the health care effect on animals is enhanced.
Preferably, the mesoporous silica is pretreated by the following raw materials in parts by weight: 6-10 parts of mesoporous silica, 15-20 parts of ammonium persulfate solution, 20-30 parts of deionized water, 9-12 parts of dodecylamine and 15-20 parts of ethanol solution.
By adopting the technical scheme, the mesoporous silica can be modified by ammonium persulfate, and has more oxygen-containing groups, so that dodecylamine is grafted, the hydrophobicity of the mesoporous silica is enhanced, the saturated precipitation amount in an organic solvent is increased, and the encapsulation amount of the mesoporous silica in zein is improved.
Preferably, the schizochytrium limacinum powder is pretreated by the following steps: mixing the schizochytrium limacinum powder with deionized water and an antioxidant, homogenizing, adding porous starch, oscillating in a water bath at 30-35 ℃ for adsorption for 60-70min, adding beta-cyclodextrin and epichlorohydrin, regulating the pH to 9-10, heating to 50-55 ℃, stirring for 5-6h, regulating the pH to be neutral, centrifuging, washing with ethanol and deionized water in sequence, and drying to obtain the treated schizochytrium limacinum powder.
Because DHA is a long-chain unsaturated fatty acid, the physical and chemical properties of the DHA are active, and the DHA is extremely easy to be influenced by factors such as illumination, heat, sample and the like to generate oxidation reaction, the DHA is likely to be influenced by external environmental factors such as sunlight, oxygen environment high temperature and the like in the storage process to generate oxidation reaction, further the quality of products is influenced, and in addition, the peroxide generated by DHA oxidation has certain toxicity, so the peroxide of the schizochytrium limacinum powder containing DHA is controlled; by adopting the technical scheme, after the schizochytrium limacinum powder is mixed with deionized water and an antioxidant, porous starch containing a loose porous structure is added to adsorb the schizochytrium limacinum powder and the antioxidant, the antioxidant has an inhibition effect on the oxidation reaction of the schizochytrium limacinum powder, and the porous starch also has a certain protection effect on the schizochytrium limacinum powder, so that the schizochytrium limacinum powder is stored in the pores of the porous starch and is not easily influenced by factors such as sunlight and oxygen to oxidize; in addition, the strong hydrophilicity of the porous starch also severely restricts the adsorption force of the porous starch on mesoporous silica with a hydrophobic component, so that epoxy chloropropane is added as a cross-linking agent to enable the porous starch and beta-cyclodextrin to undergo a cross-linking reaction, so that the thermal stability of the porous starch is improved, and the porous starch has a good adsorption effect on hydrophobic substances, so that the porous starch can be tightly adsorbed with the mesoporous silica when the schizochytrium limacinum powder is loaded in the mesoporous silica.
Preferably, when the schizochytrium limacinum powder is pretreated, the following raw materials in parts by weight are adopted: 2.5-3 parts of schizochytrium limacinum powder, 1-1.5 parts of deionized water, 0.05-0.2 part of antioxidant, 5-8 parts of porous starch, 20-30 parts of beta-cyclodextrin and 0.5-1 part of epichlorohydrin.
By adopting the technical scheme, the dosage of the porous starch of the adsorbent is reasonably controlled, so that the adsorbent has high adsorption quantity and good oxidation resistance, and the porous starch is reacted with a proper amount of cross-linking agent and beta-cyclodextrin, so that the stability of the porous starch is improved, the adsorption effect of the porous starch on the mesoporous silica of the hydrophobic substance is improved, the loading stability of the schizochytrium powder in the mesoporous silica is improved, and the structural stability of the mesoporous silica loaded schizochytrium powder in the rumen is improved.
Preferably, the antioxidant comprises dandelion extract and corn silk extract in a mass ratio of 1-3:0.5-2.
By adopting the technical scheme, the dandelion extract and the corn silk extract contain flavonoid compounds, the oxidation resistance is high, toxic peroxide generated by the oxidation of schizochytrium limacinum powder can be prevented, the dandelion extract and the corn silk extract have the effects of clearing heat and detoxicating, detumescence and resolving hard mass and promoting milk secretion, and the milk yield can be improved when the feed is used for feeding ruminants.
In a second aspect, the present application provides a rumen bypass schizochytrium limacinum powder, which adopts the following technical scheme:
a rumen bypass schizochytrium limacinum powder is prepared by a preparation method of rumen bypass schizochytrium limacinum powder.
By adopting the technical scheme, the prepared rumen bypass schizochytrium limacinum powder has strong antioxidation capability, is not easy to oxidize by air and the like, can improve the stability of the rumen bypass schizochytrium limacinum powder in the rumen, is not decomposed in the rumen, and ensures that the release amount of the rumen bypass schizochytrium limacinum powder after the small intestine is decomposed, and the utilization rate is improved.
In summary, the present application has the following beneficial effects:
1. because mesoporous silica is adopted as a load of the schizochytrium limacinum powder, the corn protein is encapsulated on the mesoporous silica to prepare a core material under the action of supercritical carbon dioxide, and finally the core material is coated by palm oil fat powder and corn fiber glue to prepare rumen-bypass schizochytrium limacinum powder, the schizochytrium limacinum powder is coated in multiple layers, the rumen-bypass rate of the schizochytrium limacinum powder in rumen is remarkably improved, and after the schizochytrium limacinum powder enters the intestinal tract, the schizochytrium limacinum powder is decomposed in the intestinal tract, so that the effective substances of the schizochytrium limacinum powder can be released in the intestinal tract to be absorbed and converted by an organism, and the effective utilization rate of the effective substance DHA is remarkably improved.
2. In the application, the mesoporous silica is preferably pretreated by ammonium persulfate, dodecylamine and the like, so that the hydrophobicity of the mesoporous silica is enhanced, the solubility of the mesoporous silica in an organic solvent is reduced, and therefore, when supercritical carbon dioxide is diffused into the solvent to expand the solvent, the supersaturation precipitation amount of the mesoporous silica is increased, the encapsulation rate of zein is improved, and the stability of the schizochytrium limacinum powder is improved.
3. In the application, porous starch is preferably used for loading the schizochytrium limacinum powder, and the antioxidant is added into the schizochytrium limacinum powder, so that the DHA (docosahexaenoic acid) in the schizochytrium limacinum powder is not easy to be influenced by light, heat, oxygen and other environmental factors, and the stability of the effective substances in storage and use can be ensured.
Detailed Description
Examples
The following raw materials are commercially available in the examples, in which the following raw materials are used as examples, and the schizochytrium limacinum powder is selected from the company of the Sihe Iceland Biotechnology, model BH-LHZF; the mesoporous silica is selected from Nanjing Xianfeng nano material technology Co., ltd, and the product number is XFF29; the zein is selected from Shandong Xin such as chemical industry Co., ltd, model 140; the palm oil fat powder is selected from Nantong sand and chemical technology limited company, and the model is high-energy treasure II-type; the porous starch is selected from the biological technology limited company of Xian Chen Hui Kang Ze, and the product number is T4214; beta-cyclodextrin is selected from the group consisting of flay biotechnology limited, su state, with the product number 4201; the herba Taraxaci extract is selected from SiamiTONGZO Biotechnology Co., ltd, and has a product number of TZ18166; the corn silk extract is selected from Shaanxi Sail Biotechnology Co., ltd, and has the product number of SR0027.
Example 1: a preparation method of rumen bypass schizochytrium limacinum powder comprises the following steps:
s1, preparing a core material: (1) Uniformly mixing 1kg of schizochytrium limacinum powder with deionized water according to a mass ratio of 1:3, adding 4kg of mesoporous silica, carrying out ultrasonic treatment at room temperature for 30min, heating to 37 ℃, stirring for 40min, centrifuging, and drying to prepare the mesoporous silica-loaded schizochytrium limacinum powder;
(2) Uniformly mixing mesoporous silica-loaded schizochytrium limacinum powder with 9kg of dichloromethane and 5kg of ethanol, adding 7kg of zein, uniformly mixing, introducing supercritical carbon dioxide, boosting to 8MPa, preserving heat at 45 ℃ for 90min, cooling to room temperature, drying, and preparing a core material, wherein the flow rate of carbon dioxide is 2kg/h;
s2, preparing a wall material: heating 1kg of palm oil fat powder to a molten state, dissolving 0.6kg of corn fiber glue in water, mixing with the molten state palm oil fat powder, uniformly mixing to prepare a wall material, and extracting the corn fiber glue from corn bran by an alkaline hydrogen peroxide method;
s3, coating: mixing 1kg of wall material, 0.3kg of core material and citric acid, regulating the pH value to 8, heating to 90 ℃, uniformly stirring, and coating to obtain the rumen-protected schizochytrium limacinum powder.
Example 2: a preparation method of rumen bypass schizochytrium limacinum powder comprises the following steps:
s1, preparing a core material: (1) Uniformly mixing 1kg of schizochytrium limacinum powder with deionized water according to a mass ratio of 1:5, adding 5kg of mesoporous silica, carrying out ultrasonic treatment at room temperature for 40min, heating to 40 ℃, stirring for 40min, centrifuging, and drying to prepare the mesoporous silica-loaded schizochytrium limacinum powder;
(2) Uniformly mixing mesoporous silica-loaded schizochytrium limacinum powder with 9kg of dichloromethane and 5kg of ethanol, adding 10kg of zein, uniformly mixing, introducing supercritical carbon dioxide, boosting to 10MPa, preserving heat at 55 ℃ for 100min, cooling to room temperature, drying, and preparing a core material, wherein the flow rate of the carbon dioxide is 3kg/h;
s2, preparing a wall material: heating 1kg of palm oil fat powder to a molten state, dissolving 1kg of corn fiber glue in water, mixing with the molten state palm oil fat powder, uniformly mixing to prepare a wall material, and extracting the corn fiber glue from corn bran by an alkaline hydrogen peroxide method;
s3, coating: mixing 1kg of wall material, 0.5kg of core material and citric acid, regulating the pH value to 10, heating to 95 ℃, uniformly stirring, and coating to obtain the rumen-protected schizochytrium limacinum powder.
Example 3: the preparation method of the rumen bypass schizochytrium limacinum powder is different from the embodiment 1 in that mesoporous silica is pretreated by the following steps:
(1) Adding 6kg of mesoporous silica into 15kg of ammonium persulfate aqueous solution with mass concentration of 18%, stirring at room temperature for 24 hours, centrifuging, washing the centrifugate with distilled water until the pH value is neutral, and drying to obtain oxidized mesoporous silica;
(2) Adding oxidized mesoporous silica into 20kg of deionized water, carrying out ultrasonic treatment for 30min, adding 9kg of dodecylamine and 15kg of ethanol solution, heating to 80 ℃, stirring for 24h, and drying.
Example 4: the preparation method of the rumen bypass schizochytrium limacinum powder is different from the embodiment 1 in that mesoporous silica is pretreated by the following steps:
(1) Adding 10kg of mesoporous silica into 20kg of ammonium persulfate aqueous solution with mass concentration of 20%, stirring at room temperature for 20h, centrifuging, washing the centrifugate with distilled water until the pH value is neutral, and drying to obtain oxidized mesoporous silica;
(2) Adding oxidized mesoporous silica into 30kg of deionized water, carrying out ultrasonic treatment for 20min, adding 12kg of dodecylamine and 20kg of ethanol solution, heating to 90 ℃, stirring for 20h, and drying.
Example 5: a preparation method of rumen bypass schizochytrium limacinum powder is different from example 4 in that step (2) is not performed during mesoporous silica pretreatment.
Example 6: a preparation method of rumen bypass schizochytrium limacinum powder is different from example 4 in that step (1) is not carried out when mesoporous silica is pretreated, mesoporous silica is added into 20kg of deionized water, ultrasound is carried out for 30min, 9kg of dodecylamine and 15kg of ethanol solution are added, the temperature is raised to 80 ℃, stirring is carried out for 24h, and drying is carried out.
Example 7: the preparation method of the rumen bypass schizochytrium limacinum powder is different from the embodiment 4 in that the schizochytrium limacinum powder is pretreated by the following steps: mixing 2.5kg of schizochytrium limacinum powder with 1kg of deionized water and 0.05kg of antioxidant, homogenizing, adding 5kg of porous starch, carrying out water bath oscillation adsorption for 70min at 30 ℃, adding 20kg of beta-cyclodextrin and 0.5kg of epichlorohydrin, regulating the pH value to 9, heating to 50 ℃, stirring for 5h, regulating the pH value to be neutral, centrifuging, washing with ethanol and deionized water in sequence, and drying to obtain the treated schizochytrium limacinum powder, wherein the antioxidant is dandelion extract and corn silk extract with the mass ratio of 1:0.5.
Example 8: the preparation method of the rumen bypass schizochytrium limacinum powder is different from the embodiment 4 in that the schizochytrium limacinum powder is pretreated by the following steps: mixing 3kg of schizochytrium limacinum powder with 1.5kg of deionized water and 0.2kg of antioxidant, homogenizing, adding 8kg of porous starch, oscillating in a water bath at 35 ℃ for adsorption for 60min, adding 30kg of beta-cyclodextrin and 1kg of epichlorohydrin, regulating the pH to 10, heating to 55 ℃, stirring for 6h, regulating the pH to be neutral, centrifuging, washing with ethanol and deionized water in sequence, and drying to obtain the treated schizochytrium limacinum powder, wherein the antioxidant is dandelion extract and corn silk extract with the mass ratio of 3:2.
Example 9: a preparation method of rumen bypass schizochytrium limacinum powder is different from example 8 in that beta-cyclodextrin and epichlorohydrin are not added.
Example 10: a preparation method of rumen bypass schizochytrium limacinum powder is different from example 8 in that no porous starch is added.
Example 11: a preparation method of rumen bypass schizochytrium limacinum powder is different from example 8 in that no antioxidant is added.
Example 12: a preparation method of rumen bypass schizochytrium limacinum powder is different from example 8 in that equivalent amount of corn silk extract is used to replace dandelion extract.
Comparative example
Comparative example 1: a preparation method of rumen bypass schizochytrium limacinum powder is different from example 1 in that no wall material is added.
Comparative example 2: a preparation method of rumen bypass schizochytrium limacinum powder is different from that of the embodiment 1 in that corn fiber gum is not added in the wall material.
Comparative example 3: the preparation method of the rumen bypass schizochytrium limacinum powder is different from the embodiment 1 in that the core material is prepared by the following method: uniformly mixing 1kg of schizochytrium limacinum powder with 3kg of deionized water, adding 4kg of mesoporous silica and 7kg of zein, uniformly mixing, and drying to prepare the core material.
Comparative example 4: a schizochytrium limacinum powder is selected from Xiean Bingham Biotechnology Limited liability company, and has model BH-LHZF.
Comparative example 5: a preparation method of Rumex protective schizochytrium limacinum powder comprises sieving Cera flava and schizochytrium limacinum powder with 40 mesh sieve, heating 550kg Cera flava to melt, controlling the melting temperature at 70deg.C, adding into a high-speed shearing mixer granulator together with 300kg schizochytrium limacinum powder and 0.4kg natural compound antioxidant, and mixing at 400r/min; the stirring speed is increased to 800r/min during high-speed shearing and granulating, and simultaneously, the shearing slurry slow gear is started for 5min; circulating cooling water with the temperature of 8 ℃ is input into the heating and cooling sleeve, particles are obtained after cooling to the room temperature, the prepared particles pass through a 16-layer vibrating screen, and the materials in the screen are collected to be the first-level coated particles; 140kg of cellulose acetate phthalate and 9.6kg of glycerol monoacetate are dissolved in 200L of organic solvent (the volume ratio is 1:1 of ethanol and acetone mixture), and the prepared coating material is uniformly sprayed on the surfaces of the primary coating particles in a bottom jet fluidized bed, wherein the spraying pressure is 0.2MPa, the spraying speed is 10L/min, the air inlet temperature is 40 ℃, and the time is 20min; and (3) passing the two layers of coated particles through a 12-mesh analysis screen to obtain the rumen bypass schizochytrium limacinum powder product with the schizochytrium limacinum powder content of 30 percent.
Performance test
1. Rumen bypass effect detection: the schizochytrium limacinum powder was prepared according to the methods of examples 1-12 and comparative examples 1-5, and the degradation rate of the schizochytrium limacinum powder in the rumen was examined with reference to the following method, and the examination results are recorded in table 1.
1. Test animals: the rumen bypass chytrid powder prepared in examples 1 to 12 and comparative examples 1 to 5 was tested on 4 Holstein cows fitted with permanent rumen fistulae as test subjects, and the results are shown in Table 1 below, and the data in Table 1 are the average of four replicates;
2. the test method comprises the following steps: respectively putting a plastic pipe with a nylon bag fixed into rumen sacs of all test cattle through rumen fistulae at one time 1 hour before feeding in the morning, fixing the plastic pipe at the mouth of the fistulae by nylon wires on the plastic pipe, starting timing, detecting rumen degradation rate of rumen-passing schizochytrium powder in 2h, 4h, 6h, 12h and 24h, and filling rumen-passing schizochytrium powder into the nylon bags in a blank test, and flushing for 2-3min until water is clear;
the rumen degradation rate was calculated using the following formula: rumen degradation rate (%) = degradation amount of rumen bypass chytrid powder at a certain culture point/weight of correction bagging bypass chytrid powder 100%;
wherein the degradation amount of the rumen bypass chytrid at a certain time point = the weight of the correction bagging rumen bypass chytrid-the weight of the rumen bypass chytrid residue at a certain time point;
correction bag weight by-rumen schizochytrium powder = actual bagging weight by-rumen schizochytrium powder (1-by-rumen schizochytrium powder bagging escape rate);
the rumen bypass chytrid bagging escape rate (%) = (weight of rumen bypass chytrid in blank test-weight of rumen bypass chytrid residue in blank test)/weight of rumen bypass chytrid 100% of blank test.
Table 1 degradation rate of rumen bypass schizochytrium limacinum powder measured by nylon bag method
In the embodiment 1 and the embodiment 2, the schizochytrium limacinum powder, the mesoporous silica and the zein are adopted as core materials, the palm fat powder and the corn fiber glue are adopted as wall materials, and the rumen bypass schizochytrium limacinum powder prepared by supercritical carbon dioxide has small degradation rate in rumen, so that after the schizochytrium limacinum powder is coated by the mesoporous silica and the wall materials, the active ingredient DHA in the schizochytrium limacinum powder is effectively protected, the release of the schizochytrium limacinum powder in the rumen is relieved, and the rumen bypass effect is good.
In examples 3 and 4, mesoporous silica was pretreated with ammonium persulfate, dodecylamine, etc., so as to enhance the hydrophobicity of the mesoporous silica and reduce the solubility of the mesoporous silica, and as can be seen from the data in table 1, the degradation rate of the rumen-bypass schizochytrium powder prepared in examples 3 and 4 in the rumen is reduced compared with example 1, and the protective effect of the mesoporous silica on the schizochytrium powder is enhanced, which means that the pretreated mesoporous silica can reduce the degradation rate of the schizochytrium powder in the rumen.
In example 5, compared with example 4, when mesoporous silica was pretreated, the oxidized mesoporous silica was not treated with dodecylamine, and the hydrophobicity of the mesoporous silica was reduced, and as shown in table 1, the degradation rate of the rumen bypass schizochytrium in the rumen prepared in example 5 was greater than that of example 4, indicating that improving the hydrophobicity of the mesoporous silica can enhance the protection of the schizochytrium in the rumen and reduce the degradation rate of the rumen bypass schizochytrium in the rumen.
Compared with the embodiment 4, the embodiment 6 has the advantages that mesoporous silica is not pretreated by ammonium persulfate, dodecyl amine is directly used for treatment, the degradation speed of the rumen bypass schizochytrium powder prepared in the embodiment 6 in the rumen is increased, the degradation rate is increased, the protection force of mesoporous silica on the schizochytrium powder is weakened, the fact that the mesoporous silica is oxidized by ammonium persulfate first, and then the stability of the mesoporous silica loaded schizochytrium powder in the rumen can be improved by connecting with long carbon chain alkylamine, and the degradation rate of the schizochytrium powder in the rumen is reduced.
Examples 7 and 8 the schizochytrium powder was further pretreated with porous starch, antioxidant, etc. on the basis of example 4, and the rumen bypass schizochytrium powder prepared in examples 7 and 8 was further reduced in the degradation rate in the rumen and improved in the rumen bypass effect as compared with example 4.
Example 9 compared with example 8, the rumen degradation rate of the rumen bypass schizochytrium limacinum powder prepared in example 9 is increased and the rumen bypass effect is reduced as compared with the example 8, wherein beta-cyclodextrin and epichlorohydrin are not used for pretreatment of the schizochytrium limacinum powder, which shows that the rumen bypass effect of the schizochytrium limacinum powder can be improved by using the beta-cyclodextrin and the epichlorohydrin for pretreatment of the schizochytrium limacinum powder.
In example 10, compared with example 8, porous starch was not added when the schizochytrium limacinum powder was pretreated in example 10, and the data in table 1 show that the rumen bypass schizochytrium limacinum powder prepared in example 10 has an increased degradation rate, which may be that the schizochytrium limacinum powder cannot be supported on the porous starch and the connection stability with mesoporous silica cannot be improved, so that the schizochytrium limacinum powder has an increased degradation rate in the rumen.
In example 11, compared with example 8, no antioxidant was added when the schizochytrium limacinum powder was pretreated, in example 12, the dandelion extract was replaced with the corn silk extract in an equivalent amount, and in the rumen bypass schizochytrium limacinum powder prepared in examples 11 and 12, the degradation rate in the rumen was similar to that in example 8.
In comparative example 1, the core material is coated by using no wall material, compared with example 1, the degradation rate of the schizochytrium limacinum powder in the rumen is obviously increased, and the degradation rate of the schizochytrium limacinum powder in the rumen is increased to 45.24% after 24 hours, so that the schizochytrium limacinum powder is degraded quickly in the rumen; in comparative example 2, corn fiber gum was not added to the wall material, so that the degradation rate of the rumen bypass schizochytrium limacinum powder was increased and the rumen bypass effect was reduced as compared with example 1.
Comparative example 3 is different from example 1 in that the degradation rate of the prepared rumen bypass chytrid powder is increased and the rumen bypass effect is reduced by directly mixing the raw materials when preparing the core material.
Comparative example 4 is a commercial schizochytrium limacinum powder, which is degraded by 13.85% only for 2 hours in the rumen, and has fast degradation speed and poor rumen bypass effect.
Comparative example 5 is rumen bypass schizochytrium limacinum powder prepared in the prior art, and the degradation speed in the rumen is faster than that of example 1, and the rumen bypass effect is inferior to that of example 1.
2. Lactation yield and DHA content detection: the schizochytrium limacinum powder was prepared according to the methods of examples 1-12 and comparative examples 1-5, and the milk yield of cows and the DHA content in the milk were examined with reference to the following methods, and the examination results are recorded in Table 2.
1. Test animals: 180 Holstein cows with similar ages, birth times, body conditions and milk yields are equally divided into 18 groups; 2. the test method comprises the following steps: feeding basic ration in groups 1-12, and adding 50g of the rumen bypass schizochytrium limacinum powder prepared in examples 1-12 every day; comparative examples 1-5 groups were fed with basal diet, and 50g of the rumen bypass schizochytrium limacinum powder prepared in comparative examples 1-5 was added per day; group 18 was a control group, fed basal daily ration daily. The test cows are fed by a single groove for 3 times in the morning, in the middle and at night, milking is carried out for 3 times, rumen bypass schizochytrium limacinum powder is mixed in concentrate, the pre-feeding period is 10 days, the formal period is 30 days, and the test period is 40 days; milk yields at days 10, 20, 30 and 40 of the test period were measured and 50mL of cow milk was collected at an early, medium and late ratio of 4:3:3 for DHA content measurement of milk.
TABLE 2 detection of milk yield and DHA content in cow milk
The cows of examples 1-12 and comparative examples 1-5 had similar milk yields on days 10, 20, 30 and 40, but the DHA content of the cows of examples 3 and 4 was increased as compared to example 1, the DHA content of the cows of examples 7 and 8 was further increased as compared to example 4, and the DHA content of the cows of examples 5 and 6 was reduced as compared to example 4, which means that the DHA content of the cows of examples 7 and 8 was the highest, indicating that the DHA degradation in the rumen of the schizochytrium powder of examples 7 and 8 was less and could be mostly absorbed and converted. In the comparative example 1, the wall material is not coated, DHA in the schizochytrium limacinum powder is easily degraded and destroyed in the rumen, the DHA content in the cow milk is obviously reduced, and in the comparative example 2, corn fiber glue is not added in the wall material, and the DHA content in the cow milk in the comparative example 2 is not as same as that in the example 1; the cows of comparative examples 3-5 had similar yields, but had reduced DHA content compared to example 1, and no DHA was detected in the cow milk when the control group was fed the basal diet.
3. Storage stability detection: the rumen bypass schizochytrium powder prepared in the examples and the comparative examples is exposed to the air at the same time, and 10 volunteers are selected to evaluate the smell of the rumen bypass schizochytrium powder when the rumen bypass schizochytrium powder is placed for 10 days, 20 days and 30 days, and the rumen bypass schizochytrium powder is scored according to the following standard, wherein the fishy smell is very serious in 1-3 points; 4-6 minutes of slight fishy smell; the results of each group of test, which were averaged over 10 volunteers and reported in Table 3, were free of fishy smell for 7-10 minutes.
TABLE 3 shelf stability detection of rumen bypass schizochytrium limacinum powder
| Group of | Day 10 | Day 20 | Day 30 |
| Example 1 | 7.8 | 6.4 | 5.8 |
| Example 2 | 7.6 | 6.8 | 6.0 |
| Example 3 | 8.5 | 7.5 | 6.9 |
| Example 4 | 8.6 | 7.8 | 7.0 |
| Example 5 | 8.2 | 7.3 | 6.7 |
| Example 6 | 8.4 | 7.2 | 6.6 |
| Example 7 | 9.5 | 8.9 | 8.4 |
| Example 8 | 9.8 | 9.2 | 8.5 |
| Example 9 | 8.9 | 8.4 | 8.0 |
| Example 10 | 8.7 | 8.3 | 7.8 |
| Example 11 | 8.8 | 8.2 | 7.6 |
| Example 12 | 8.9 | 8.4 | 7.7 |
| Comparative example 1 | 5.6 | 4.2 | 2.4 |
| Comparative example 2 | 6.8 | 5.9 | 3.7 |
| Comparative example 3 | 7.5 | 5.4 | 4.2 |
| Comparative example4 | 4.5 | 3.2 | 1.2 |
| Comparative example 5 | 6.9 | 5.4 | 3.1 |
As can be seen from the data in table 3, the rumen bypass schizochytrium limacinum powder prepared in example 1 and example 2 has a score of between 4 and 6 minutes after being placed at normal temperature for 30 days, has slight fishy smell, and indicates that DHA is partially oxidized; the mesoporous silica of the embodiment 3 and the embodiment 4 is pretreated, the fishy smell evaluation score is increased after being placed for 30 days, the fishy smell is reduced, and the antioxidation effect of the schizochytrium limacinum powder can be improved after the mesoporous silica is pretreated; in examples 5 and 6, dodecylamine and ammonium persulfate were not used respectively in the pretreatment of mesoporous silica, and table 3 shows that the rumen bypass schizochytrium powder prepared in examples 5 and 6 has a fishy smell score lower than that of example 4 after being placed for 30 days, which indicates that the oxidation amount of DHA is increased; example 7 and example 8 showed an increase in the fishy smell score of the rumen bypass schizochytrium limacinum powder after 30 days of standing, compared with example 4, indicating an increase in the oxidation resistance of the rumen bypass schizochytrium limacinum powder prepared in examples 7-8.
Example 9 compared to example 8, beta-cyclodextrin and epichlorohydrin were not used when the schizochytrium limacinum powder was pretreated, and the rumen bypass schizochytrium limacinum powder prepared in example 9 was stored for 30 days, and the fishy smell score was higher than that of example 8 and the storage stability was reduced.
In example 10, compared with example 8, porous starch was not added when the schizochytrium limacinum powder was pretreated in example 10, the rumen bypass schizochytrium limacinum powder prepared in example 10 was increased in oxidation amount, decreased in oxidation resistance, and increased in fishy smell.
In examples 11 to 12, compared with example 8, no antioxidant was added when the schizochytrium limacinum powder was pretreated, in example 12, the equivalent amount of corn silk extract was used instead of dandelion extract, the rumen bypass schizochytrium limacinum powder prepared in examples 11 and 12 was more fishy smell than example 8 after 30 days of standing, the fishy smell was increased, and the oxidation resistance of the schizochytrium limacinum powder was decreased.
In comparative example 1, the core material was not coated with the wall material, whereas in comparative example 2, corn fiber gum was not added to the wall material, the amount of oxidized rumen bypass schizochytrium limacinum powder was increased, and the fishy smell was severe, as compared with example 1.
Comparative example 3 is different from example 1 in that the raw materials are directly mixed at the time of preparing the core material, and the prepared rumen bypass schizochytrium limacinum powder has a smell score similar to that of example 1 at 10 days, but has a fishy smell increased and a score significantly decreased at 20 days and 30 days.
Comparative example 4 is a commercial schizochytrium limacinum powder, the fishy smell score is lower on day 10, the fishy smell is heavier, comparative example 5 is a rumen bypass schizochytrium limacinum powder prepared by the prior art, no obvious fishy smell is generated on day 10 of storage, but the fishy smell is gradually increased with the extension of time, and the fishy smell is very serious on day 30.
The present embodiment is merely illustrative of the present application and is not intended to be limiting, and those skilled in the art, after having read the present specification, may make modifications to the present embodiment without creative contribution as required, but is protected by patent laws within the scope of the claims of the present application.
Claims (9)
1. The preparation method of the rumen bypass schizochytrium limacinum powder is characterized by comprising the following steps of:
preparing a core material: (1) Uniformly mixing the schizochytrium limacinum powder with deionized water, adding mesoporous silicon dioxide, carrying out ultrasonic treatment at room temperature for 30-40min, heating to 37-40 ℃, uniformly stirring, centrifuging, and drying to obtain mesoporous silicon dioxide loaded schizochytrium limacinum powder;
(2) Uniformly mixing mesoporous silica-loaded schizochytrium limacinum powder with dichloromethane and ethanol, adding zein, uniformly mixing, introducing supercritical carbon dioxide, boosting to 8-10MPa, preserving heat at 45-55 ℃ for 90-100min, cooling to room temperature, and drying to obtain a core material;
preparing a wall material: heating palm oil fat powder to a molten state, dissolving corn fiber glue in water, mixing with the molten state palm oil fat powder, and uniformly mixing to prepare a wall material;
coating: mixing the wall material, the core material and the citric acid, regulating the pH value to 8-10, heating to 90-95 ℃, uniformly stirring, and coating to obtain rumen bypass schizochytrium limacinum powder;
the mesoporous silica is pretreated by the following steps:
(1) Adding mesoporous silica into ammonium persulfate aqueous solution with mass concentration of 18-20%, stirring at room temperature for 20-24h, centrifuging, washing the centrifugate with distilled water until the pH value is neutral, and drying to obtain oxidized mesoporous silica;
(2) Adding the mesoporous silica into deionized water, carrying out ultrasonic treatment for 20-30min, adding dodecylamine and ethanol solution, heating to 80-90 ℃, stirring for 20-24h, and drying.
2. The method for preparing the rumen bypass schizochytrium limacinum powder according to claim 1, which is characterized in that:
the mass ratio of the core material to the wall material is 0.3-0.5:1;
in the core material, the mass ratio of the schizochytrium limacinum powder to the mesoporous silica to the zein is 1:4-5:7-10;
in the wall material, the mass ratio of the palm oil fat powder to the corn fiber gum is 1:0.6-1.
3. The preparation method of the rumen bypass schizochytrium limacinum powder according to claim 1, wherein the mass ratio of the schizochytrium limacinum powder to deionized water in the step (1) is 1:3-5;
the mass ratio of dichloromethane to ethanol to zein in the step (2) is 9:5:7-10.
4. The method for preparing the rumen bypass schizochytrium limacinum powder according to claim 1, characterized in that the flow rate of carbon dioxide is 2-3kg/h.
5. The preparation method of the rumen bypass schizochytrium limacinum powder according to claim 1, wherein the mesoporous silica is prepared from the following raw materials in parts by weight: 6-10 parts of mesoporous silica, 15-20 parts of ammonium persulfate solution, 20-30 parts of deionized water, 9-12 parts of dodecylamine and 15-20 parts of ethanol solution.
6. The method for preparing the rumen bypass schizochytrium limacinum powder according to claim 1, wherein the schizochytrium limacinum powder is pretreated by the following steps: mixing the schizochytrium limacinum powder with deionized water and an antioxidant, homogenizing, adding porous starch, oscillating in a water bath at 30-35 ℃ for adsorption for 60-70min, adding beta-cyclodextrin and epichlorohydrin, regulating the pH to 9-10, heating to 50-55 ℃, stirring for 5-6h, regulating the pH to be neutral, centrifuging, washing with ethanol and deionized water in sequence, and drying to obtain the treated schizochytrium limacinum powder.
7. The preparation method of the rumen bypass schizochytrium limacinum powder according to claim 6, wherein when the schizochytrium limacinum powder is pretreated, the following raw materials are in parts by weight: 2.5-3 parts of schizochytrium limacinum powder, 1-1.5 parts of deionized water, 0.05-0.2 part of antioxidant, 5-8 parts of porous starch, 20-30 parts of beta-cyclodextrin and 0.5-1 part of epichlorohydrin.
8. The preparation method of the rumen bypass schizochytrium limacinum powder according to claim 6, wherein the antioxidant comprises dandelion extract and corn silk extract in a mass ratio of 1-3:0.5-2.
9. A rumen bypass schizochytrium limacinum powder, characterized in that it is prepared by the preparation method of the rumen bypass schizochytrium limacinum powder according to any one of claims 1-8.
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