TW202000037A - Compound liposome additive of soy lecithin-coated vitamin and preparing method thereof capable of effectively enhancing growth of chicken - Google Patents

Abstract

Provided is a compound liposome additive of soy lecithin-coated vitamin, which includes four kinds of liposomes of vitamin A, D3, E and K3 coated with lecithin. The weight ratio of lecithin to the vitamin A is preferably to be 2:0.1, the weight ratio of lecithin to the vitamin D3 is preferably to be 2:0.5, the weight ratio of lecithin to the vitamin E is preferably to be 2:0.4, and the weight ratio of lecithin to the vitamin K3 is preferably to be 2:1. The method of preparing the compound liposome additive includes the steps of: (a) formulating lecithin and ethanol into a lecithin ethanol aqueous solution; (b) mixing vitamins with the aforementioned aqueous solution according to the weight ratios thereof; (c) concentrating under reduced pressure different aqueous solutions, respectively, so as to form a thin film; (d) adding pure water or ethanol to the thin film and shaking the same until the thin film is completely melted in the water; (e) shaking respectively to obtain vitamin A, D3, E and K3 liposome aqueous solutions of different concentrations; and (f) mixing to form a compound liposome additive.

Description

Soy lecithin-coated vitamin compound lipid particle additive and preparation method thereof

The invention relates to a compound lipid particle additive, in particular to a compound lipid particle poultry food additive in which vitamins are coated with soybean lecithin.

Vitamins are necessary to maintain the physiological functions of chickens and other poultry. If the intake of vitamins in chicken feed is not uniform, it may cause disease and economic consequences. The current vitamin deficiency in poultry can be classified into the following main causes:

1. Improper feed ratio: due to the different vitamin content in the feed, for example, corn contains vitamin A, soybean cake (meal) contains more vitamin E, and bone meal contains more vitamin B12. Therefore, when the feed ratio is not uniform or the poultry prefers a specific feed, some vitamins will be too high and another vitamin will be excreted, which will lead to insufficient intake of vitamins by the poultry. Therefore, how to maintain the balance of vitamins is required for the ratio of feed and drinking water. Matters needing attention.

2. Processing and storage methods: Many vitamins have a time effect during feed storage and feed processing after production, which can lead to vitamin instability. Therefore, pay attention to the time effect when using feed or adding vitamins. In addition, vitamins are also susceptible to temperature, humidity, sunlight, acid and alkali.

3. Abuse of antibiotics: Due to market factors, farmers use antibiotics in large quantities, which in turn leads to inadequate microbial synthesis of vitamins in the gut of poultry. The addition of vegetable oils and animal fats to feeds not only improves the daily concentration, but also reduces the digestive chyme throughput rate and thus enhances nutrient absorption, solving the problem of insufficient vitamin synthesis in the poultry gut. Generally, the weak fat emulsifying ability in the early growth stage of poultry leads to the weak digestion ability of fat. At this time, if the emulsifier can be used to improve the emulsifying effect, the utilization rate of poultry fat can be improved. Because soybean lecithin contains linoleic acid, linolenic acid and other unsaturated fatty acids, soy lecithin added to the feed can promote fat metabolism, decomposition and digestion and can absorb excess cholesterol, increase the lean meat rate of poultry (Li Man, 2011). The study found that lecithin coated with vitamin B6 can delay the release in chicken crops (Heba Abd El Azim, et al, 2015). Therefore, lecithin can not only be added to the feed as a nutrient component, such as added to the liquid feed can contribute to the stability of emulsification, when used as a coating material, lecithin can also increase the permeability of the coating.

Although lecithin also has emulsifying properties to prevent cholesterol from depositing on the walls of blood vessels, and emulsified lipids can enhance the body's immune function, and can promote the phagocytic function of phagocytes and lymphocyte proliferation, but its main source is high-purity lecithin price high. Products with a lecithin content of less than 55% are usually used in health foods or nutritional products, or as an adjunct to pharmaceutical products. Products with a lecithin content of 60 to 80% are usually used in cosmetics or as pharmaceutical excipients. Products with a lecithin content of more than 90% are usually used in the pharmaceutical industry, and the higher the purity, the stronger the oxidizing performance. It needs to be hydrogenated and then stored. The non-hydrogenated high-purity lecithin should be stored in a nitrogen-filled sealed container. Indirectly, it also increases the cost of holding.

Animal lecithin, such as egg yolk, contains 73% of egg yolk lipids. Its saturation range is better but the price is higher. Soy lecithin contains about 29% to 39% of vegetable lecithin. Soy lecithin contains about 29% to 39% of the lecithin. The purity is not high but the price is relatively low. If it can be refined with low-purity food lecithin, vitamin liposomes can be prepared. Should be able to significantly reduce costs. At present, although the development technology of nanomaterials has been widely used in the application of food additives and food-related ingredients, it is less common as an application in the development of agriculture and feed. Many related literatures have not discussed the use of low-purity food lecithin packages. Relevant researches on the use purity of lecithin covered with vitamins A, D ₃ , E, K ₃ , the use ratio of coated vitamins, vitamin concentration, etc. Therefore, the development of nano-composite materials for compound lipid vitamins in feed additives will be the future development trend.

The main purpose of the present invention is to provide a compound lipid particle supplement of soybean lecithin coated vitamins that can effectively help chickens grow.

Another object of the present invention is to provide a compound lipid particle supplement of highly stable soybean lecithin-coated vitamins.

Another object of the present invention is to provide a method for preparing a compound liposome additive with the above-mentioned effects.

In order to achieve the above object, a preferred embodiment of the present invention includes four liposomes coated with vitamin A, D3, E, and K3 with lecithin, characterized in that the weight ratio of the lecithin to the vitamin A is Is 2:0.01 to 2:0.1, the weight ratio of the lecithin to the vitamin D3 is 2:0.2 to 2:0.5, the weight ratio of the lecithin to the vitamin E is 2:0.2 to 2:0.4, the lecithin The weight ratio with this vitamin K3 is 2:0.4 to 2:1.

In some embodiments, a more specific range may be a weight ratio of the lecithin to the vitamin A of 2:0.1, a weight ratio of the lecithin to the vitamin D3 of 2:0.5, a weight ratio of the lecithin to the vitamin E The weight ratio is 2:0.4, and the weight ratio of the lecithin to the vitamin K3 is 2:1.

In some embodiments, the particle sizes of the four liposomes are all between 123.3±0.5 nm and 319.98±44.8 nm, so as to have good stability.

The preparation method of the compound lipid particle additive preferably includes the following steps: (a) disposing the lecithin and ethanol into a 2% aqueous solution of lecithin ethanol for use; (b) according to the vitamin A, D3, E, K3 The weight ratio is mixed with the 2% aqueous lecithin ethanol solution to form different concentrations of vitamin lecithin ethanol aqueous solution; (c) the different concentrations of the vitamin lecithin ethanol aqueous solution are concentrated under reduced pressure to form films; (d) the films Add pure water or ethanol separately, shake with a shaker until all the film is dropped into the water, and obtain a multi-layered liposome solution; (e) shake these multi-layered liposome solutions with an ultrasonic cell shaker to obtain different concentrations Vitamin A, D3, E, K3 liposome aqueous solution; (f) Mix these different concentrations of vitamin A, D3, E, K3 liposome aqueous solution to form a compound liposome additive.

In some embodiments, before step (b), the vitamins A, D3, E, and ethanol may be formulated into an aqueous solution of vitamins A, D3, and E, respectively, and the vitamin K3 and pure water may be formulated into vitamin K3. The aqueous solution is then mixed with the 2% ethanol solution of lecithin.

In some embodiments, the lecithin in step (a) is extracted with low-purity food lecithin with a purity of about 30% using acetone and ethanol to obtain the lecithin with a purity of more than 70%.

In some embodiments, the method for extracting lecithin includes the following steps: (g) mixing low-purity food lecithin with a purity of about 30% and acetone at a weight ratio of 1:2, stirring with an electromagnetic heating stirrer, and then standing Remove the supernatant; (h) Repeat step (g) twice; (i) After the last stirring, perform suction filtration to take the precipitate; (j) The precipitate and the purity of 95% ethanol 1:2 weight Proportional mixing, stirring using an electromagnetic heating stirrer, suction filtration is performed immediately after each stirring; (k) The solution obtained after filtration is concentrated under reduced pressure to completely remove the solvent to obtain the lecithin with a purity of more than 70%.

In some embodiments, the concentrations of the vitamin A, D3, E and K3 liposome aqueous solutions obtained in step (e) are 0.5%, 0.25%, 0.2% and 1%, respectively.

Vitamins are divided into fat-soluble vitamins and water-soluble vitamins. Vitamin A can help poultry visual function, reproduction function, immune function; vitamin E can achieve antioxidant, protect the integrity of biofilm, improve egg production rate, fertilization rate, hatchability rate; vitamins D3 promotes the absorption of calcium and phosphorus in the gastrointestinal tract and improves the immune function; when the poultry lacks vitamin K, it is easy to bleed and not easy to solidify. Therefore, based on the above factors, the present invention uses a particulate carrier to coat the vitamin A, D3, E, and K3 materials, and then prepares a comprehensive vitamin particle carrier-containing poultry beverage additive, which can prevent the problem of uneven intake of vitamins in poultry diets. .

The invention provides a compound lipid particle supplement of soy lecithin coated vitamins, which mainly comprises four kinds of lipid particles coated with vitamin A, D3, E and K3 by lecithin.

Vitamins are divided into fat-soluble vitamins and water-soluble vitamins. Vitamin A can help poultry visual function, reproduction function, immune function; vitamin E can achieve antioxidant, protect the integrity of biofilm, improve egg production rate, fertilization rate, hatchability rate; vitamins D3 promotes the absorption of calcium and phosphorus in the gastrointestinal tract and improves the immune function; when the poultry lacks vitamin K, it is easy to bleed and not easy to solidify. Therefore, based on the above factors, the invention uses lecithin to coat the vitamin A, D3, E and K3 materials, and then prepares a comprehensive vitamin microparticle-containing poultry drink additive, which can prevent the problem of uneven intake of vitamins in poultry diets. .

Liposomes are mainly hollow spheres formed by the accumulation of phospholipids in lecithin. When phospholipids are dispersed in water, they will automatically form a double-layer membrane structure. Experiments have shown that liposomes have good biocompatibility and promote growth of osteoblast-like cells (Huang, et al, 1997); liposomes are classified according to particle size and number of lipid bilayers, and can be divided into single Layer of small liposomes (SUV's), single layer of large liposomes (LUV's) and multi-layered liposomes (MLV's).

Referring to FIG. 1, FIG. 1 shows a flow chart of the method for preparing a compound liposome particle supplement of soybean lecithin-coated vitamins of the present invention. In order to make the compound lipid particle supplement of vitamin A, D3, E, K3 provided by the soybean lecithin provided by the present invention into a product with good stability and stability, the present invention provides a preparation in a preferred embodiment The method of the compound lipid particle additive mentioned above mainly includes the following steps: (a) Lecithin and ethanol are configured as a lecithin ethanol aqueous solution for use; first, high-purity (more than 70%) lecithin and ethanol are mixed to a 2% concentration The ethanol solution of lecithin is ready for use. (b) Vitamins A, D3, E, and K3 are mixed with an aqueous solution of lecithin ethanol according to their weight ratio; the vitamins A, D3, E, and K3 are mixed with the aqueous solution of 2% concentration of lecithin ethanol according to their weight ratio, respectively , To form different concentrations of vitamin lecithin ethanol aqueous solution. (c) Different concentrations of vitamin lecithin ethanol aqueous solutions were concentrated under reduced pressure to form thin films; different concentrations of these vitamin lecithin ethanol aqueous solutions were concentrated under reduced pressure to form thin films. (d) Add the film to pure water or ethanol and shake until the film is fully incorporated into the water to obtain a multi-layered lipid solution; add these films to pure water or ethanol and shake with a oscillating machine until all the film falls into the water and obtain a multi-layered lipid Grain solution. (e) Ultrasonic shattering of the multi-layer liposome solution to obtain aqueous solutions of vitamin liposomes with different concentrations; these multi-layer liposome solutions were shaken with an ultrasonic cell shaker to obtain vitamin A, D3, E. K3 liposome aqueous solution. (f) Mix different concentrations of vitamin liposome aqueous solution to form a compound liposome additive. The vitamin A, D3, E, K3 liposome aqueous solutions of different concentrations are mixed to form a compound liposome additive.

The contents of the above steps are described in detail as follows:

In step (a), lecithin and ethanol are configured as a lecithin ethanol aqueous solution for standby, and 2 g of lecithin is added to 99.5% ethanol for quantitative measurement to 100 mL, and configured as a 2% lecithin ethanol aqueous solution for standby. Among them, lecithin can directly use high-purity (more than 70%) lecithin, but due to the high price of high-purity (more than 70%) lecithin, you can also choose to use the commercially available low-purity low-purity (70 %) Lecithin is extracted and prepared. The method of extraction and preparation will be described in detail later.

Step (b) Vitamins A, D3, E, and K3 are mixed with an aqueous solution of lecithin ethanol according to their weight ratio; the vitamins A, D3, E, and K3 are mixed with the aqueous solution of 2% concentration of lecithin ethanol according to their weight ratio, respectively Mix to form ethanol solution of vitamin lecithin with different concentrations.

Among them, the vitamin A lecithin ethanol aqueous solution is 100ml of the above 2% lecithin ethanol aqueous solution and 0.01%~0.1% vitamin A is poured into the eggplant type bottle, the vitamin A can be mixed with ethanol to prepare 0.01%~0.1% vitamin A 100mL of ethanol solution, then pour into eggplant type bottle together with 100mL of 2% lecithin ethanol aqueous solution.

Among them, the vitamin D3 lecithin ethanol aqueous solution is poured into the eggplant-shaped bottle with the above 2% lecithin ethanol aqueous solution 100mL and 0.2%~0.5% vitamin D3. The vitamin D3 can be mixed with ethanol to prepare 0.2%~0.5% vitamin D3 100mL of ethanol solution, then pour into eggplant type bottle together with 100mL of 2% lecithin ethanol aqueous solution.

Among them, the vitamin E lecithin ethanol aqueous solution is 100ml of the above 2% lecithin ethanol aqueous solution and 0.2%~0.4% vitamin E is poured into the eggplant type bottle, the vitamin E can be mixed with ethanol to prepare 0.2%~0.4% vitamin E 100mL of ethanol solution, then pour into eggplant type bottle together with 100mL of 2% lecithin ethanol aqueous solution.

Step (c) Different concentrations of vitamin lecithin ethanol aqueous solution are concentrated under reduced pressure to form a thin film; the different concentrations of these vitamin lecithin ethanol aqueous solutions are reduced at a reduced pressure concentrator speed of 110 rpm and a water bath temperature of 37±1° C. Press to concentrate for about 10~15 minutes to remove the organic solvent, remove it after forming a film.

Step (d) Add the film to pure water or ethanol and shake until the film is fully incorporated into the water to obtain a multi-layer lipid solution; add these films to pure water or ethanol respectively and shake with a oscillating machine until all the film falls into the water and obtain a multi-layer micro-lipid Lipid solutions (MLVs).

Among them, the film of vitamin A was poured into 200mL of pure water, and shaken with a test tube shaker until all the film fell into the water.

Among them, the film of vitamin D3 was poured into 200mL of pure water, and shaken with a test tube shaker until all the film fell into the water.

Among them, the film of vitamin K3 was poured into 200mL of pure water, and shaken with a test tube shaker until all the film fell into the water.

Among them, because vitamin E is insoluble in water, the film of vitamin E is poured into 200 mL of 25% ethanol water, and shaken with a test tube shaker until all the film falls into the water.

Step (e) Ultrasonic shredding of the multi-layered liposome solution to obtain aqueous solutions of vitamin liposomes with different concentrations; after shaking these multi-layered liposome solutions with an ultrasonic cell shaker, respectively, to obtain vitamins A and D3 with different concentrations , E, K3 liposome aqueous solution. First, shake with a water bath ultrasonic cleaner at 700w for 20 to 40 minutes to make the vitamin liposome water more uniform, and then use a probe-type ultrasonic cell crusher to shake at 35w for 30 minutes (every 10 minutes, the outer pot of water Perform water exchange to avoid the increase of shock temperature), and finally obtain an aqueous solution of vitamin lipid particles with different concentrations of single-layer vitamin fat particles (SUV).

Among them, the finally obtained aqueous solution of vitamin A liposomes is preferably an aqueous solution of 0.5% vitamin A liposomes.

Among them, the finally obtained aqueous solution of vitamin D3 liposomes is preferably an aqueous solution of 0.25% vitamin D3 liposomes.

Among them, the finally obtained aqueous solution of vitamin E liposomes is preferably a 0.2% aqueous solution of vitamin E liposomes.

Among them, the finally obtained aqueous solution of vitamin K3 liposomes is preferably an aqueous solution of vitamin K3 liposomes with a concentration of 1%.

Step (f) Mix aqueous solutions of vitamin liposomes with different concentrations to form a compound liposome supplement. The vitamin A, D3, E, K3 liposome aqueous solutions of different concentrations are mixed to form a compound liposome additive.

In the present invention, in order to make lecithin coat vitamin A, D3, E, K3 and other materials with good stability, the following tests were conducted. In step (b) of the foregoing preparation method, the weight ratio of vitamins A, D3, E, and K3 mixed with the aqueous lecithin ethanol solution is based on the results obtained by the following tests.

When conducting the experiment, first take 2g of high-purity (purity above 70%) lecithin and add 99.5% ethanol to 100mL, configure it as a 2% lecithin ethanol aqueous solution for standby, and prepare different concentrations of vitamin ethanol solution for preparation according to its grouping, the composition is different Proportioned vitamin liposome formula, its vitamin A, D ₃ and E formula ratios are shown in Tables 1, 2, and 3, while vitamin K ₃ is configured using pure water, and its formula ratio is shown in Table 4.

Table 1 Microbiotic A liposome formula ratio table

Table 2 Probiotics D ₃ liposome formula ratio table

表3微生素E微脂粒配方比例表

Table 3 Microbial E E liposome formula ratio table

Table 4 Probiotics K3 liposome particle formulation ratio table

Stability test of vitamin liposomes:

According to Tables 1, 2, 3, and 4, configure liposome vitamins with different formulation ratios, observe whether there is oil slick, layering or precipitation in 30 days, the results show that vitamin A, D3, E, K3 liposomes Stability ratio is different.

Referring to the following table, Table 5 is the stability assessment table for different concentrations of vitamin A. Considering the ratio of the configuration of 2% lecithin ethanol solution to 0.2% vitamin A ethanol solution, stratification has occurred on the 20th day, after 30 days of stability The evaluation proves that there is no stratification of vitamin A liposomes below 0.1%, so the preparation of vitamin A liposomes at a concentration of 0.01% to 0.1% vitamin A in ethanol is determined to be a highly stable soybean lecithin-coated vitamin The compound liposome additive.

Table 5 Stability assessment table of different concentrations of vitamin A

Referring to the following table, Table 6 is the stability assessment table for different concentrations of vitamin D3. Considering the ratio of the configuration of 2% lecithin ethanol solution to 0.6% vitamin D3 ethanol solution, stratification has occurred on the 30th day, so 0.2%~ The preparation of vitamin D3 liposomes with a concentration of 0.5% vitamin D3 in ethanol was determined to be a compound liposome additive with high stability of soybean lecithin-coated vitamins.

Table 6 Stability assessment table for different concentrations of vitamin D3

Referring to the following table, Table 7 is the stability assessment table for different concentrations of vitamin E. During the configuration of vitamin E, when only pure water was used as the shaking solvent, it was found that the lecithin vitamin E ethanol aqueous film could not be shaken and blended, so 5% was prepared separately , 10%, 15%, 20%, 25% different concentrations of ethanol water production, the results show that 25% ethanol water shaking into the best effect, and in order to make the ultrasonic washing machine shake twice for a total of 40 minutes. Among them, the ratio of the concentration of the 0.6% vitamin E ethanol aqueous solution has been stratified on the 20th day, so the concentration of the vitamin E ethanol aqueous solution was reduced to 0.4% for the production. The ratio of the lecithin ethanol aqueous solution to the vitamin E ethanol aqueous solution is 2:0.4. Stable formula ratio. After 30 days of stability evaluation, the experiment proved that 0.4% vitamin E liposomes have no stratification. Therefore, the preparation of vitamin E liposomes at a concentration of 0.2% to 0.4% vitamin E ethanol solution is determined to be made. Soy lecithin with high stability is coated with vitamin liposome supplement.

Table 7 Stability assessment table for different concentrations of vitamin E

Refer to the following table. Table 8 is the stability assessment table for different concentrations of vitamin K3. Vitamin K3 is a synthetic vitamin and is not soluble in ethanol. Therefore, when prepared, it is dissolved in pure water first and added when shaking the membrane of aqueous lecithin ethanol solution. The stable ratio of phospholipid ethanol aqueous solution and vitamin K3 water is 2: 1. The ratio of the configuration of 2% lecithin ethanol aqueous solution and 1% vitamin K3 aqueous solution has stratified phenomenon on the 30th day, so 0.4%~1% vitamin K3 The preparation of vitamin K ₃ liposomes with the concentration of ethanol aqueous solution was determined to be a compound liposome additive with high stability of soybean lecithin-coated vitamins.

Table 8 Stability assessment table for different concentrations of vitamin K3

It can be known from the above test data that the soy lecithin-coated vitamin compound liposome supplement of the present invention includes four liposomes coated with vitamin A, D3, E, and K3 with lecithin, and their lecithin The weight comparison with the vitamin A is preferably 2:0.01 to 2:0.1, the weight comparison between the lecithin and the vitamin D3 is preferably 2:0.2 to 2:0.5, and the weight comparison between the lecithin and the vitamin E is preferably 2 :0.2 to 2:0.4, the weight of the lecithin and the vitamin K3 is preferably 2:0.4 to 2:1. The weight ratio of the lecithin to the vitamin A is 2:0.1, the weight ratio of the lecithin to the vitamin D3 is 2:0.5, and the weight ratio of the lecithin to the vitamin E is 2:0.4. When the weight ratio of the lecithin to the vitamin K3 is 2:1, the compounding ratio with good stability and high vitamin content can be obtained.

Referring to FIGS. 3 to 6, FIG. 3 shows the penetration microscopy observation test chart of the successful application of lecithin in the compound liposome particle supplement of the present invention, and FIG. 4 shows the successful lecithin in the compound liposome particle supplement of the present invention The penetrating microscope observation and detection diagram of coated vitamin D3. FIG. 5 shows the penetrating microscope observation and detection diagram of successfully coating vitamin E with lecithin in the compound liposome supplement of the present invention. Fig. 6 shows the penetration microscopy observation detection diagram of the vitamin K3 successfully coated with lecithin in the compound lipid particle supplement of the present invention. The lipid particles have different effects on lipid changes due to the different concentration and method of the prepared components. The present invention conducts experiments to configure different vitamin concentrations and 2% lecithin ethanol aqueous solution to observe the lipid particle size, and uses a micro pipette to obtain the prepared lipid 200ul of the vitamin solution sample was quantitatively added with 10mL of pure water for dilution, dripped into the dilution solution on the copper mesh, lyophilized and taken out for observation through a transmission microscope, and the experiment proved that the method for preparing compound liposome additives of the present invention, All can successfully coat vitamin A, D3, E and K3.

Referring to FIG. 2, it shows a flow chart of the method for extracting lecithin in the compound lipid particle supplement of the present invention. The lecithin used in the present invention can be extracted with acetone and ethanol using low-purity food lecithin with lower purity and lower price to obtain the lecithin with a purity of more than 70%. The low-purity food lecithin used in the present invention is preferred in this embodiment, but not limited to food lecithin with a purity of about 30%, which is relatively inexpensive, and the low-purity food lecithin referred to in the present invention should contain a purity of less than 70% The food lecithin.

Among them, the method for extracting lecithin is preferably carried out in the following steps: (g) Mix and stir low-purity soybean lecithin and acetone in a weight ratio of 1:2; mix low-purity food lecithin and acetone according to a purity of about 30% 1:2 weight ratio mixing, use an electromagnetic heating stirrer to stir and then stand to remove the supernatant. (h) Repeat step (g) twice. (i) After the last stirring, perform suction filtration to take the precipitate. (j) The precipitate is mixed with ethanol at a weight ratio of 1:2 and stirred using a stirrer. After stirring, the solution is suctioned and filtered to obtain a solution; the precipitate is mixed with a purity of 95% ethanol at a weight ratio of 1:2 and an electromagnetic heating stirrer is used Stir, perform suction filtration immediately after each stirring. (k) Repeat step (j) twice, and remove the solvent by vacuum concentration to obtain lecithin with a purity of more than 70%; repeat the ethanol extraction step (j) twice, and obtain the solution after the last filtration, and then decompress Concentration removes the solvent completely to obtain the lecithin with a purity of more than 70%.

For the extraction method, refer to the solvent extraction method. First, the commercially available low-purity food lecithin and acetone are stirred with a magnet using an electromagnetic heating stirrer at a ratio of 1:2 for 30 minutes. After standing for 30 minutes, the supernatant is removed, and then this step is used. Repeat twice. After the last stirring, perform suction filtration, take the precipitate into the beaker, and then use the electromagnetic heating stirrer to stir the magnet for 30 minutes according to the ratio of food lecithin: 95% ethanol = 1:2. Perform the same steps one to four times, and perform suction filtration immediately after each stirring. After filtration, the solution is concentrated under reduced pressure (110rpm, 37±1℃, time about 2 hours), and the solvent can be completely removed to obtain refined lecithin. , Then weighed to calculate the extraction rate.

In the present invention, the commercially available low-purity food lecithin is extracted three times with acetone and 95% ethanol three times. The purity of the lecithin can be increased to 73.7 ± 2.2%, and the recovery rate is 84.8 ± 4.6%.

In addition, the particle size analysis of the liposomes was performed on the compound lipid particle supplement of soy lecithin-coated vitamins prepared by the method of the present invention. The analysis method was to analyze the nanostructured lipid carrier with a particle size analyzer (Malvern Zetasizer Nano ZS). Particle size and polydispersity index (Polydispersity index, PDI) distribution value. When measuring the particle size, first put the sample tank into the instrument and measure it repeatedly three times. Finally, repeat the measurement results of the experiment to find the average and standard deviation.

The lipid particles have different concentrations and methods of the prepared components, which affect the lipid changes. In the present invention, different vitamin concentrations and a 2% phospholipid ethanol solution are arranged during the analysis, and the particle size of the lipid particles is observed. Referring to the following Table 9 to Table 12, the liposome vitamin A, D3, E, K3 formula ratio of 2:0.1, 2:0.5, 2:0.4, 2:1 to prepare vitamin liposome, the four groups of samples were weighed Covered under four different conditions of 23±2℃, 4℃, 45℃, 4℃/45℃, after observing the stability of liposomes for 30 days, it was found that vitamin A liposomes changed at 23±2℃ and 4 ℃ The stability of the particle size will not change significantly, but the stability of vitamin D3, E, K3 liposomes at 4 ℃ will not change significantly. The particle size of the above liposomes is about 123.3±0.5 nm To 319.98±44.8 nm. From the above observation results, it is confirmed that the stability of the vitamin-coated liposomes made by the present invention will not change significantly.

Table 9 Comparison of properties of liposomes of vitamin A at different temperatures

Table 10 Comparison of properties of liposomes of vitamin D _{3 at} different temperatures

Table 11 Comparison of properties of liposomes of vitamin E at different temperatures

Table 12 Comparison of properties of liposomes of vitamin K _{3 at} different temperatures

The invention refers to the general broiler feeding method and uses the traditional captive feeding method to perform the effectiveness evaluation and analysis. The effectiveness evaluation and analysis are weighed once every 8 days, and each group is synchronously fed for 35 days. Group A is a vitamin aqueous solution made of the compound lipid particle additive of soybean lecithin-coated vitamins of the present invention. Group B vitamin water is prepared by adding 10g of food-grade emulsifier at the concentration of group A vitamin aqueous solution, and Group A and Group B are diluted 1000 times in the same proportion to the concentration of commercially available formula. 500mL is added to drinking water once a day in the chick stage (1-10 days), and medium-age chickens in the hair removal stage (11-25 days) are added daily 1000mL to drinking water once a day, adult chickens (26~35 days) drink a higher amount of water. US and Japan add 1000mL to drinking water twice a day, both provide drinking water at 6:30 every day, and stop water at 22:30 the night before, To ensure that the chickens are drinking water.

Referring to Table 13, it was found that the average weight of chickens in the compound vitamin lipid particles of the present invention was 2264.1±609.5 g, and the feed conversion rate was 4.9%, while the average weight of chickens in the vitamin aqueous solution group was 2093.5±380.3 g. The feed conversion rate is 5.2%. The experimental results show that the combination of vitamin A, D ₃ , E, and K ₃ with liposomes can effectively help chickens grow.

Table 13 Average poultry body weight n=16, Mean±SD

The present invention has been disclosed using the above preferred embodiments, but it is not intended to limit the present invention. Those with ordinary knowledge in the technical field to which the present invention pertains should clearly understand that the present invention is not limited to the details of the above illustrative embodiments. The present invention can be implemented in other specific forms without departing from the basic spirit of the present invention. The embodiments are merely illustrative The present invention does not limit the present invention. The present invention is based on the scope of patent application, rather than the above description. All the modifications and meanings in the scope of patent application are within the scope of the present invention.

a~f‧‧‧step g~k‧‧‧step

FIG. 1 shows a flow chart of the method for preparing a compound liposome particle supplement of soybean lecithin-coated vitamins of the present invention. FIG. 2 shows a flow chart of the method for extracting lecithin from the compound liposome particle supplement of the present invention. FIG. 3 shows a transmission microscope observation and detection diagram of the lecithin in the compound liposome supplement of the present invention successfully coated with vitamin A. FIG. Fig. 4 shows the penetration microscopy observation and detection diagram of the lecithin in the compound liposome supplement of the present invention successfully coated with vitamin D3. FIG. 5 shows the penetration microscopy observation and detection diagram of the lecithin in the compound liposome supplement of the present invention successfully coated with vitamin E. FIG. Fig. 6 shows the penetration microscopy observation detection diagram of the vitamin K3 successfully coated with lecithin in the compound lipid particle supplement of the present invention.

a~f‧‧‧step

Claims (8)

A soy lecithin-coated vitamin compound lipid particle supplement, comprising four kinds of lipid particles coated with vitamin A, D3, E, and K3 with lecithin, characterized by the weight of the lecithin and the vitamin A The ratio is 2:0.01 to 2:0.1, the weight ratio of the lecithin to the vitamin D3 is 2:0.2 to 2:0.5, the weight ratio of the lecithin to the vitamin E is 2:0.2 to 2:0.4, the egg The weight ratio of phospholipid to the vitamin K3 is from 2:0.4 to 2:1. The compound liposome additive as described in item 1 of the patent application scope, wherein the weight ratio of the lecithin to the vitamin A is 2:0.1, and the weight ratio of the lecithin to the vitamin D3 is 2:0.5, the The weight ratio of lecithin to the vitamin E is 2:0.4, and the weight ratio of the lecithin to the vitamin K3 is 2:1. The compound lipid particle additive as described in item 1 of the patent application scope, wherein the particle size of the four lipid particles is between 123.3±0.5 nm and 319.98±44.8 nm. A method for preparing the compound lipid particle additive according to any one of the items 1 to 3 of the patent application scope, which comprises the following steps: (a) disposing the lecithin and ethanol into a 2% aqueous solution of lecithin ethanol for use ; (B) The vitamins A, D3, E, K3 are mixed with the 2% aqueous lecithin ethanol solution according to their weight ratio to form different concentrations of vitamin lecithin ethanol aqueous solution; (c) The vitamin lecithin ethanol of different concentrations The aqueous solution was concentrated under reduced pressure to form thin films; (d) The films were added to pure water or ethanol, and shaken with a shaker until all the films were dropped into the water to obtain multiple layers of liposome solution; (e) The multiple layers of microlipid solution The lipid particle solution was shaken with an ultrasonic cell shaker, respectively, to obtain different concentrations of vitamin A, D3, E, K3 liposome aqueous solution; (f) mixing these different concentrations of vitamin A, D3, E, K3 liposome The aqueous solution forms a compound liposome additive. The preparation method as described in item 4 of the patent application scope, wherein before step (b), the vitamins A, D3 and E are mixed with ethanol to prepare an aqueous solution of vitamins A, D3 and E in ethanol, and the vitamin K3 and pure water are formulated into an aqueous solution of vitamin K3, and then mixed with the 2% aqueous solution of lecithin ethanol. The preparation method as described in item 4 of the patent application scope, wherein the lecithin in step (a) is to extract low-purity food lecithin with acetone and ethanol to obtain the lecithin with a purity of more than 70%. The preparation method as described in item 6 of the patent application scope, wherein the method for extracting lecithin includes the following steps: (g) mixing the low-purity food lecithin with acetone in a weight ratio of 1:2 and stirring with an electromagnetic heating stirrer After standing, remove the supernatant; (h) Repeat step (g) twice; (i) After the last stirring, perform suction filtration to take the precipitate; (j) The precipitate and the purity of 95% ethanol 1 : 2 weight ratio mixing, stirring using an electromagnetic heating stirrer, suction filtration is performed immediately after each stirring; (k) The solution obtained after filtration is concentrated under reduced pressure, and the solvent is completely removed to obtain the lecithin with a purity of 70% or more. The preparation method as described in item 4 of the patent application scope, wherein the concentrations of the vitamin A, D3, E and K3 liposome aqueous solutions obtained in step (e) are 0.5%, 0.25% and 0.2%, respectively And 1%.

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