CN114208948A - Liposome for feed additive and preparation method thereof - Google Patents
Liposome for feed additive and preparation method thereof Download PDFInfo
- Publication number
- CN114208948A CN114208948A CN202111354261.XA CN202111354261A CN114208948A CN 114208948 A CN114208948 A CN 114208948A CN 202111354261 A CN202111354261 A CN 202111354261A CN 114208948 A CN114208948 A CN 114208948A
- Authority
- CN
- China
- Prior art keywords
- volatile oil
- liposome
- rhizoma atractylodis
- mixed solution
- probiotics
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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- 239000002502 liposome Substances 0.000 title claims abstract description 85
- 239000003674 animal food additive Substances 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000000341 volatile oil Substances 0.000 claims abstract description 140
- 239000006041 probiotic Substances 0.000 claims abstract description 49
- 235000018291 probiotics Nutrition 0.000 claims abstract description 49
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 36
- 239000000203 mixture Substances 0.000 claims abstract description 32
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- 239000000463 material Substances 0.000 claims abstract description 30
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- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims abstract description 18
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Classifications
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Abstract
The invention discloses a liposome for a feed additive, which belongs to the technical field of livestock and poultry feed and comprises a liposome formed by lecithin and cholesterol, and a volatile oil composition and probiotics encapsulated in the liposome; wherein the volatile oil composition comprises cinnamon volatile oil, houttuynia cordata volatile oil and rhizoma atractylodis volatile oil. The invention also discloses a preparation method of the liposome, which comprises the steps of extracting cinnamon, houttuynia cordata and rhizoma atractylodis through ultrasonic-assisted distillation to obtain cinnamon volatile oil, houttuynia cordata volatile oil and rhizoma atractylodis volatile oil; and then taking lecithin and cholesterol as wall materials, and encapsulating the volatile oil composition and the probiotics to obtain the liposome. The liposome improves the stability and bioavailability of the volatile oil and the probiotics, is convenient to store and transport, and can be used as a feed additive to play the drug effects of the volatile oil and the probiotics.
Description
Technical Field
The invention relates to the technical field of livestock and poultry feed, in particular to a liposome for a feed additive and a preparation method thereof.
Background
The feed additive is a small amount or trace substance added in the production, processing and using processes of feed, and the feed additive is small in dosage but remarkable in effect. The feed additive is a raw material inevitably used in modern feed industry, and has obvious effects on strengthening the nutritive value of basic feed, improving the production performance of animals, ensuring the health of the animals, saving the feed cost, improving the quality of animal products and the like.
With the occurrence of avian influenza and the like, huge losses are brought to animal breeding owners. In order to reduce risks, the breeding owners often use antibiotics in large dose in an illegal way; in order to improve the yield, the hormone is used in large dose. The long-term or large-scale use of antibiotics causes a large amount of antibiotic residues in edible products such as livestock and poultry meat, milk and the like, poisoning or allergy of edible people occurs occasionally, and the residual hormone can cause side effects such as osteoporosis, low immune function, premature maturity of children and the like of the eaters.
The natural product contains various nutrient components and bioactive substances, has double functions of nutrition and medicines (antimicrobial, antiparasitic, anti-inflammatory and analgesic, etc.), can improve the production performance and prevent diseases, and has the characteristics of low residue, no pollution, difficult generation of drug resistance, etc. However, the natural plant extracts are generally liquid substances which are difficult to dissolve in water, if the natural plant extracts are directly added into the feed as feed additives without treatment, the natural plant extracts are not beneficial to storage and transportation of the feed, the bioavailability of the feed can be greatly reduced, the exertion of the drug effect is limited, meanwhile, the stability of the extracts is poor, and under the action of oxygen, illumination and heat, active ingredients are easy to decompose or deteriorate, so that the quality and the drug effect of the product are seriously influenced.
Disclosure of Invention
Based on the above, the invention provides the liposome for the feed additive and the preparation method thereof, the cinnamon medicinal material, the houttuynia cordata medicinal material and the rhizoma atractylodis medicinal material are extracted to obtain the volatile oil composition, and then the volatile oil composition and the probiotics are encapsulated in the liposome, so that the stability and the bioavailability of the volatile oil and the probiotics are improved, the storage and the transportation are convenient, and meanwhile, the liposome is used as the feed additive to exert the drug effects of the volatile oil and the probiotics.
The technical scheme adopted by the invention is as follows:
a liposome for feed additive and its preparation method comprise liposome formed by lecithin and cholesterol, and volatile oil composition and probiotic bacteria encapsulated in the liposome;
wherein the volatile oil composition comprises cinnamon volatile oil, houttuynia cordata volatile oil and rhizoma atractylodis volatile oil.
In the liposome for feed additive disclosed herein, the probiotic is a mixture of yeast, bacillus, bifidobacterium, lactobacillus rhamnosus and lactobacillus acidophilus.
In the liposome for feed additive disclosed in the present application, the ratio of lecithin: the mass ratio of the cholesterol is 3-5: 1.
In the liposome for feed additive disclosed in the present application, the liposome: volatile oil composition: the mass ratio of the probiotics is 10-20: 4: 1.
In the liposome for feed additive disclosed herein, the cinnamon essential oil: houttuynia volatile oil: the mass ratio of the rhizoma atractylodis volatile oil is 2-3: 1-2: 1.
The preparation method of the liposome for the feed additive comprises the following steps:
s1, extracting volatile oil
Drying and pulverizing cortex Cinnamomi, herba Houttuyniae and rhizoma Atractylodis respectively, and steam distilling to obtain cortex Cinnamomi volatile oil, herba Houttuyniae volatile oil and rhizoma Atractylodis volatile oil;
s2, preparation of liposome
S21, weighing probiotics, dissolving the probiotics in a PBS buffer solution to obtain a mixed solution 1, and keeping the temperature at 40-50 ℃ for later use;
s22, weighing cinnamon volatile oil, houttuynia cordata volatile oil, rhizoma atractylodis volatile oil, lecithin, cholesterol and an emulsifier, and dissolving the materials in a proper amount of chloroform-ethanol mixed solution to obtain a mixed solution 2;
s23, putting the mixed solution 2 into a water bath for heating, carrying out reduced pressure rotary evaporation, removing the solvent in the mixed solution 2, and drying to obtain a layer of uniform lipid membrane;
s24, adding the mixed solution 1 into a lipid membrane, and carrying out ultrasonic mixing at 60-80W for 20-40 min to obtain a suspension; emulsifying the suspension under high pressure, and filtering to obtain liposome;
s25, adding a freeze-drying protective agent into the liposome, freezing at-70 to-100 ℃, and then carrying out freeze-drying to obtain the liposome freeze-dried powder.
In the preparation method disclosed in the present application, the step S1 specifically includes: crushing the dried cinnamon medicinal material, the dried houttuynia cordata medicinal material and the dried rhizoma atractylodis medicinal material by using a crusher to obtain medicinal material powder; weighing medicinal powder, adding 5-10 times of distilled water, and carrying out ultrasonic treatment at 35-40 ℃ under 300-400W for 20-30 min; distilling and extracting the solution after ultrasonic treatment at 100-120 ℃ for 3-5 h, collecting oily substances in the distillation condensate, adding a proper amount of anhydrous sodium sulfate for dehydration, and filtering with a microporous filter membrane to obtain cinnamon volatile oil, houttuynia cordata volatile oil and rhizoma atractylodis volatile oil.
In the preparation method disclosed in the present application, the step S23 specifically includes: and (3) putting the mixed solution 2 into a water bath, carrying out reduced pressure rotary evaporation at 40-50 ℃ for 20-40 min, removing the solvent in the mixed solution 2, and drying by using a vacuum drying oven to obtain a layer of uniform lipid membrane.
In the preparation method disclosed in the present application, the emulsifier in step S22 is sodium cholate.
In the preparation method disclosed in the present application, the lyoprotectant in step S25 is one or more of trehalose, mannitol, sucrose and lactose.
The cinnamon volatile oil is a volatile component extracted from dried cinnamon bark, has a strong aromatic smell, and mainly comprises aromatic aldehydes, aromatic esters, fatty acid and mushroom compounds, wherein the content of the cinnamic aldehyde is the highest. The cinnamaldehyde has antibacterial, antioxidant, and antiviral effects. The cinnamaldehyde has obvious inhibition and killing effects on various bacteria and fungi, can directly kill viruses, and can also inhibit the replication of the viruses. The cinnamaldehyde has the functions of promoting the growth of animals, improving the utilization rate of the feed and regulating and controlling intestinal flora, and can increase the fragrance of the feed and attract the animals to eat.
The houttuynia cordata is a plant of the Saururaceae, belongs to perennial herbaceous plants, and is a medicine and a food. The houttuynia volatile oil is a main functional substance, has the effects of resisting bacteria, disinfecting, clearing heat, relieving pain, enhancing immunity of organisms and promoting urination, can be used as a feed for livestock, plays an important role in preventing and treating diseases of the livestock, and has a good application effect.
The rhizoma atractylodis is a plant of the genus atractylodis of the family Compositae, and research shows that the rhizoma atractylodis volatile oil has the functions of broad-spectrum antibiosis, antivirus, anti-inflammation, gastrointestinal movement function adjustment, bone calcification promotion and the like. The addition of the rhizoma atractylodis volatile oil in the feed has good effect on preventing infectious bronchitis, laryngitis and the like of poultry.
The probiotics are bacteria beneficial to animals, play a beneficial role by improving the ecological balance of host intestinal flora after being put into an organism, and have the effects of preventing or improving diarrhea, preventing infection of a reproductive system, enhancing the immunity of the organism, promoting the health of an intestinal digestive system, helping to absorb nutrient components and the like. The probiotic bacteria have reduced activity during storage and intestinal digestion, and are entrapped in the liposome to help protect against the invasion of gastric acid, bile and digestive enzymes.
The invention has the beneficial effects that:
extracting cortex Cinnamomi, herba Houttuyniae and rhizoma Atractylodis by ultrasonic-assisted distillation to obtain cortex Cinnamomi volatile oil, herba Houttuyniae volatile oil and rhizoma Atractylodis volatile oil. And lecithin and cholesterol are taken as wall materials, the volatile oil and the probiotics are encapsulated to form liposome, so that the stability and bioavailability of the volatile oil and the probiotics are improved, the storage and transportation are convenient, and the drug effect of the volatile oil and the probiotics is exerted as a feed additive. The prepared liposome is used as a feed additive and added into chicken feed, so that the diarrhea rate of the chicken can be obviously reduced.
Drawings
FIG. 1 is a graph showing the change in the total number of bacteria during the storage of liposomes in the present invention.
Detailed Description
In the following, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.
The drugs and reagents mentioned in the examples are all common commercial products unless otherwise specified.
The following provides a detailed description of embodiments of the invention.
Example 1
A liposome for feed additive comprises liposome formed from lecithin and cholesterol, and volatile oil composition and probiotic bacteria encapsulated in the liposome;
wherein the volatile oil composition comprises cinnamon volatile oil, houttuynia cordata volatile oil and atractylodes lancea volatile oil. The cinnamon volatile oil, the houttuynia cordata volatile oil and the rhizoma atractylodis volatile oil have the effects of resisting bacteria and viruses and the like, can promote the growth of animals, improve the utilization rate of feed and regulate and control the functions of intestinal flora.
The probiotic bacteria is mixture of yeast, Bacillus bifidus, Lactobacillus rhamnosus and Lactobacillus acidophilus. The probiotics are bacteria beneficial to animals, play a beneficial role by improving the ecological balance of host intestinal flora after being put into an organism, and have the effects of preventing or improving diarrhea, preventing infection of a reproductive system, enhancing the immunity of the organism, promoting the health of an intestinal digestive system, helping to absorb nutrient components and the like.
The volatile oil composition and the probiotics are encapsulated in the liposome, so that the stability and bioavailability of the volatile oil and the probiotics are improved, the storage and the transportation are convenient, and the drug effect of the volatile oil and the probiotics is exerted as a feed additive.
In a specific embodiment, the ratio of lecithin: the mass ratio of cholesterol was 3: 1.
In a specific embodiment, the liposome: volatile oil composition: the mass ratio of the probiotics is 10:4: 1.
In a specific embodiment, the ratio of cinnamon volatile oil: houttuynia volatile oil: the mass ratio of the rhizoma atractylodis volatile oil is 2:1: 1.
The preparation method of the liposome for the feed additive comprises the following steps:
s1, extracting volatile oil
Respectively crushing the dried cinnamon medicinal material, the dried houttuynia cordata medicinal material and the dried rhizoma atractylodis medicinal material by using a crusher to obtain medicinal material powder; weighing the medicinal powder, adding 5 times of distilled water, and performing ultrasonic treatment at 35 deg.C under 300W for 20 min; distilling the solution after ultrasonic treatment at 100 deg.C for 3 hr, collecting oily substance in the distillate, adding appropriate amount of anhydrous sodium sulfate for dehydration, and filtering with microporous membrane to obtain cortex Cinnamomi volatile oil, herba Houttuyniae volatile oil and rhizoma Atractylodis volatile oil. The extraction efficiency of the volatile oil can be improved by extracting the volatile oil with the assistance of ultrasonic waves.
S2, preparation of liposome
S21, weighing probiotics, dissolving the probiotics in a PBS (phosphate buffer) solution to obtain a mixed solution, and keeping the temperature at 40 ℃ for later use.
S22, weighing cinnamon volatile oil, houttuynia cordata volatile oil, rhizoma atractylodis volatile oil, lecithin, cholesterol and sodium cholate, and dissolving the materials in a proper amount of chloroform-ethanol mixed solution to obtain a mixed solution 2.
S23, putting the mixed solution 2 into a water bath, carrying out reduced pressure rotary evaporation at 40 ℃ for 20min, removing the solvent in the mixed solution 2, and drying by using a vacuum drying oven to obtain a layer of uniform lipid membrane.
S24, adding the mixed solution 1 into a lipid membrane, and carrying out ultrasonic mixing at 60W for 20min to obtain a suspension; emulsifying the suspension under high pressure, and filtering to obtain liposome.
S25, adding a freeze-drying protective agent into the liposome, freezing at-70 ℃, and then carrying out freeze-drying to obtain the liposome freeze-dried powder. The freeze-drying protective agent is one or more of trehalose, mannitol, sucrose and lactose.
The encapsulation efficiency of the liposomes of this example was 44.3% by HPLC method.
Example 2
A liposome for feed additive comprises liposome formed from lecithin and cholesterol, and volatile oil composition and probiotic bacteria encapsulated in the liposome;
wherein the volatile oil composition comprises cinnamon volatile oil, houttuynia cordata volatile oil and atractylodes lancea volatile oil. The cinnamon volatile oil, the houttuynia cordata volatile oil and the rhizoma atractylodis volatile oil have the effects of resisting bacteria and viruses and the like, can promote the growth of animals, improve the utilization rate of feed and regulate and control the functions of intestinal flora.
The probiotic bacteria is mixture of yeast, Bacillus bifidus, Lactobacillus rhamnosus and Lactobacillus acidophilus. The probiotics are bacteria beneficial to animals, play a beneficial role by improving the ecological balance of host intestinal flora after being put into an organism, and have the effects of preventing or improving diarrhea, preventing infection of a reproductive system, enhancing the immunity of the organism, promoting the health of an intestinal digestive system, helping to absorb nutrient components and the like.
The volatile oil composition and the probiotics are encapsulated in the liposome, so that the stability and bioavailability of the volatile oil and the probiotics are improved, the storage and the transportation are convenient, and the drug effect of the volatile oil and the probiotics is exerted as a feed additive.
In a specific embodiment, the ratio of lecithin: the mass ratio of cholesterol was 4: 1.
In a specific embodiment, the liposome: volatile oil composition: the mass ratio of the probiotics is 15:4: 1.
In a specific embodiment, the ratio of cinnamon volatile oil: houttuynia volatile oil: the mass ratio of the rhizoma atractylodis volatile oil is 2.5:1.5: 1.
The preparation method of the liposome for the feed additive comprises the following steps:
s1, extracting volatile oil
Respectively crushing the dried cinnamon medicinal material, the dried houttuynia cordata medicinal material and the dried rhizoma atractylodis medicinal material by using a crusher to obtain medicinal material powder; weighing the medicinal powder, adding 7 times of distilled water, and performing ultrasonic treatment at 37 deg.C under 350W for 25 min; distilling the solution after ultrasonic treatment at 110 deg.C for 4 hr, collecting oily substance in the distillate, adding appropriate amount of anhydrous sodium sulfate for dehydration, and filtering with microporous membrane to obtain cortex Cinnamomi volatile oil, herba Houttuyniae volatile oil and rhizoma Atractylodis volatile oil. The extraction efficiency of the volatile oil can be improved by extracting the volatile oil with the assistance of ultrasonic waves.
S2, preparation of liposome
S21, weighing probiotics, dissolving the probiotics in a PBS (phosphate buffer) solution to obtain a mixed solution, and keeping the temperature at 45 ℃ for later use.
S22, weighing cinnamon volatile oil, houttuynia cordata volatile oil, rhizoma atractylodis volatile oil, lecithin, cholesterol and sodium cholate, and dissolving the materials in a proper amount of chloroform-ethanol mixed solution to obtain a mixed solution 2.
S23, putting the mixed solution 2 into a water bath, carrying out reduced pressure rotary evaporation at 45 ℃ for 30min, removing the solvent in the mixed solution 2, and drying by using a vacuum drying oven to obtain a layer of uniform lipid membrane.
S24, adding the mixed solution 1 into a lipid membrane, and carrying out ultrasonic mixing at 70W for 30min to obtain a suspension; emulsifying the suspension under high pressure, and filtering to obtain liposome.
S25, adding a freeze-drying protective agent into the liposome, freezing at-85 ℃, and freeze-drying to obtain the liposome freeze-dried powder. The freeze-drying protective agent is one or more of trehalose, mannitol, sucrose and lactose.
The encapsulation efficiency of the liposome of this example was 41.7% by HPLC.
Example 3
A liposome for feed additive comprises liposome formed from lecithin and cholesterol, and volatile oil composition and probiotic bacteria encapsulated in the liposome;
wherein the volatile oil composition comprises cinnamon volatile oil, houttuynia cordata volatile oil and atractylodes lancea volatile oil. The cinnamon volatile oil, the houttuynia cordata volatile oil and the rhizoma atractylodis volatile oil have the effects of resisting bacteria and viruses and the like, can promote the growth of animals, improve the utilization rate of feed and regulate and control the functions of intestinal flora.
The probiotic bacteria is mixture of yeast, Bacillus bifidus, Lactobacillus rhamnosus and Lactobacillus acidophilus. The probiotics are bacteria beneficial to animals, play a beneficial role by improving the ecological balance of host intestinal flora after being put into an organism, and have the effects of preventing or improving diarrhea, preventing infection of a reproductive system, enhancing the immunity of the organism, promoting the health of an intestinal digestive system, helping to absorb nutrient components and the like.
The volatile oil composition and the probiotics are encapsulated in the liposome, so that the stability and bioavailability of the volatile oil and the probiotics are improved, the storage and the transportation are convenient, and the drug effect of the volatile oil and the probiotics is exerted as a feed additive.
In a specific embodiment, the ratio of lecithin: the mass ratio of cholesterol is 5:1.
In a specific embodiment, the liposome: volatile oil composition: the mass ratio of the probiotics is 20:4: 1.
In a specific embodiment, the ratio of cinnamon volatile oil: houttuynia volatile oil: the mass ratio of the rhizoma atractylodis volatile oil is 3:2: 1.
The preparation method of the liposome for the feed additive comprises the following steps:
s1, extracting volatile oil
Respectively crushing the dried cinnamon medicinal material, the dried houttuynia cordata medicinal material and the dried rhizoma atractylodis medicinal material by using a crusher to obtain medicinal material powder; weighing the medicinal powder, adding 10 times of distilled water, and performing ultrasonic treatment at 40 deg.C under 400W for 30 min; distilling the solution after ultrasonic treatment at 120 deg.C for 5 hr, collecting oily substance in the distillate, adding appropriate amount of anhydrous sodium sulfate for dehydration, and filtering with microporous membrane to obtain cortex Cinnamomi volatile oil, herba Houttuyniae volatile oil and rhizoma Atractylodis volatile oil. The extraction efficiency of the volatile oil can be improved by extracting the volatile oil with the assistance of ultrasonic waves.
S2, preparation of liposome
S21, weighing probiotics, dissolving the probiotics in a PBS (phosphate buffer) solution to obtain a mixed solution 1, and keeping the temperature at 50 ℃ for later use.
S22, weighing cinnamon volatile oil, houttuynia cordata volatile oil, rhizoma atractylodis volatile oil, lecithin, cholesterol and sodium cholate, and dissolving the materials in a proper amount of chloroform-ethanol mixed solution to obtain a mixed solution 2.
S23, putting the mixed solution 2 into a water bath, carrying out reduced pressure rotary evaporation at 50 ℃ for 40min, removing the solvent in the mixed solution 2, and drying by using a vacuum drying oven to obtain a layer of uniform lipid membrane.
S24, adding the mixed solution 1 into a lipid membrane, and carrying out ultrasonic mixing at 80W for 40min to obtain a suspension; emulsifying the suspension under high pressure, and filtering to obtain liposome.
S25, adding a freeze-drying protective agent into the liposome, freezing at-100 ℃, and then carrying out freeze-drying to obtain the liposome freeze-dried powder. The freeze-drying protective agent is one or more of trehalose, mannitol, sucrose and lactose.
The encapsulation efficiency of the liposomes of this example was 46.1% by HPLC.
Example 4
The prepared liposome is used as a feed additive for a mildew experiment
After mixing 10g of the liposome prepared in the invention in example 1, example 2 and example 3 with 2kg of feed, the mixture was placed in a constant temperature and humidity chamber with a relative humidity of 75% and a temperature of 30 +/-1 ℃, the colony growth was observed periodically by using the volatile oil composition which is not encapsulated by the liposome as a control group, and the total number of mold was measured.
The results are shown in fig. 1, and examples 1, 2 and 3 have better inhibitory effects on the growth of mold and significantly reduce the total number of mold compared to the control group. The volatile oil composition has poor stability, the active ingredients are easy to decompose or deteriorate, the antibacterial effect of the volatile oil composition is weakened along with the increase of time, the volatile oil is not easy to deteriorate after being encapsulated by the liposome, the antibacterial endurance of the volatile oil is improved, the storage is convenient, and the mildew rate of the feed additive is reduced.
Example 5
The prepared liposome serving as a feed additive is uniformly mixed with animal feed and then used for feeding animals, the effect of improving diarrhea can be realized by taking the liposome into the bodies of the animals, and the addition amount of the liposome accounts for 0.1 percent of the total mass of the animal feed when the feed is used.
The liposomes prepared in example 1, example 2 and example 3 were added to the chicken feed, 10 chicks in each group were used as an unadditized control group, 3 were repeated, and the chicks were raised for 14 days to perform experiments, and the weight change and diarrhea rate of the chicks were recorded. As shown in table 1, the addition of the liposomes prepared in examples 1, 2 and 3 reduced the diarrhea rate of the chicks compared to the control group, and example 2 showed the most significant effect.
Claims (10)
1. A liposome for feed additive is characterized in that the liposome comprises liposome formed by lecithin and cholesterol, and volatile oil composition and probiotics encapsulated in the liposome;
wherein the volatile oil composition comprises cinnamon volatile oil, houttuynia cordata volatile oil and rhizoma atractylodis volatile oil.
2. The liposome for feed additive according to claim 1, wherein the probiotic is a mixture of yeast, bacillus, bifidobacterium, lactobacillus rhamnosus and lactobacillus acidophilus.
3. The liposome for feed additive according to claim 1, wherein the ratio of lecithin: the mass ratio of the cholesterol is 3-5: 1.
4. The liposome for feed additive according to claim 1, wherein the liposome: volatile oil composition: the mass ratio of the probiotics is 10-20: 4: 1.
5. The liposome for feed additive of claim 4, wherein the cinnamon essential oil: houttuynia volatile oil: the mass ratio of the rhizoma atractylodis volatile oil is 2-3: 1-2: 1.
6. A preparation method of liposome for feed additive is characterized by comprising the following steps:
s1, extracting volatile oil
Drying and pulverizing cortex Cinnamomi, herba Houttuyniae and rhizoma Atractylodis respectively, and steam distilling to obtain cortex Cinnamomi volatile oil, herba Houttuyniae volatile oil and rhizoma Atractylodis volatile oil;
s2, preparation of liposome
S21, weighing probiotics, dissolving the probiotics in a PBS buffer solution to obtain a mixed solution 1, and keeping the temperature at 40-50 ℃ for later use;
s22, weighing cinnamon volatile oil, houttuynia cordata volatile oil, rhizoma atractylodis volatile oil, lecithin, cholesterol and an emulsifier, and dissolving the materials in a proper amount of chloroform-ethanol mixed solution to obtain a mixed solution 2;
s23, putting the mixed solution 2 into a water bath for heating, carrying out reduced pressure rotary evaporation, removing the solvent in the mixed solution 2, and drying to obtain a layer of uniform lipid membrane;
s24, adding the mixed solution 1 into a lipid membrane, and carrying out ultrasonic mixing at 60-80W for 20-40 min to obtain a suspension; emulsifying the suspension under high pressure, and filtering to obtain liposome;
s25, adding a freeze-drying protective agent into the liposome, freezing at-70 to-100 ℃, and then carrying out freeze-drying to obtain the liposome freeze-dried powder.
7. The preparation method according to claim 6, wherein the step S1 is specifically: crushing the dried cinnamon medicinal material, the dried houttuynia cordata medicinal material and the dried rhizoma atractylodis medicinal material by using a crusher to obtain medicinal material powder; weighing medicinal powder, adding 5-10 times of distilled water, and carrying out ultrasonic treatment at 35-40 ℃ under 300-400W for 20-30 min; distilling and extracting the solution after ultrasonic treatment at 100-120 ℃ for 3-5 h, collecting oily substances in the distillation condensate, adding a proper amount of anhydrous sodium sulfate for dehydration, and filtering with a microporous filter membrane to obtain cinnamon volatile oil, houttuynia cordata volatile oil and rhizoma atractylodis volatile oil.
8. The preparation method according to claim 6, wherein the step S23 is specifically: and (3) putting the mixed solution 2 into a water bath, carrying out reduced pressure rotary evaporation at 40-50 ℃ for 20-40 min, removing the solvent in the mixed solution 2, and drying by using a vacuum drying oven to obtain a layer of uniform lipid membrane.
9. The method according to claim 6, wherein the emulsifier in step S22 is sodium cholate.
10. The method according to claim 6, wherein the lyoprotectant in step S25 is one or more of trehalose, mannitol, sucrose, and lactose.
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CN115518107B (en) * | 2022-10-09 | 2023-11-03 | 北京昊坤康源医疗科学技术发展有限责任公司 | External traditional Chinese medicine preparation for treating gout and preparation method thereof |
CN115501211A (en) * | 2022-10-17 | 2022-12-23 | 蒋云云 | Traditional Chinese medicine gout plaster and preparation method thereof |
CN115501211B (en) * | 2022-10-17 | 2023-10-27 | 北京昊坤康源医疗科学技术发展有限责任公司 | Traditional Chinese medicine gout patch and preparation method thereof |
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