Chicken bone marrow phospholipid, its preparation method and application

Abstract

The application belongs to the technical field of phospholipid extraction, and particularly relates to chicken bone marrow phospholipid, a preparation method and application thereof. The method comprises the following steps: freezing and drying chicken bone marrow, mixing the chicken bone marrow in a mixed solution of dichloromethane and methanol, treating the mixture by a high-energy fluid mill, centrifuging, mixing the supernatant with water, stirring, standing, obtaining lower dichloromethane and upper mixed liquid, repeatedly extracting the upper mixed liquid to obtain a dichloromethane mixed liquid, combining the dichloromethane, reducing pressure and concentrating, drying, obtaining a mixed liposome, dissolving the mixed liposome in acetone, extracting, centrifuging, drying, and obtaining chicken bone marrow phospholipid. The antioxidant capacity of the chicken bone marrow phospholipid is further determined, which provides a theoretical basis for the application of the chicken bone marrow phospholipid in drugs for diseases caused by free radicals.

Description

Technical Field

[0001] This application belongs to the field of phospholipid extraction technology, specifically relating to a chicken bone marrow phospholipid, its preparation method, and its application. Background Technology

[0002] Phospholipids are lipid compounds containing phospholipid groups and are fundamental substances of life. They are widely found in the brain, bone marrow, liver, heart, kidneys, lungs, blood, milk, eggs, most plant seeds, and some microorganisms of animals. Phospholipids were first discovered in the human brain by Uauquelin in 1812, and later isolated from egg yolks by Gobley in 1844, naming it lecithin. Phospholipids play a significant role in activating cells, maintaining metabolism, basal metabolism, and balanced hormone secretion, as well as enhancing the body's immunity and regenerative capacity. In addition, phospholipids also promote fat metabolism, prevent fatty liver, lower serum cholesterol, improve blood circulation, and prevent cardiovascular disease. Animal lecithin mainly comes from the brain, bone marrow, liver, heart, kidneys, lungs, and blood of animals, and these sources are generally considered to be of higher quality. In contrast, plant lecithin (such as soybean lecithin) is generally a byproduct of vegetable oil production; although it is cheaper, it may pose a potential risk of allergic reactions. Currently, phospholipids are mainly derived from soybeans, with the United States and Western Europe being the world's major producers, accounting for 60% of global soybean phospholipid production. According to research statistics from DIRESAerch, the global liquid lecithin market reached 6.48 billion yuan in 2024 and is projected to reach 8.21 billion yuan by 2030, representing a compound annual growth rate (CAGR) of 4.02% from 2024 to 2030. This indicates that the global phospholipid market, especially the liquid lecithin market, is showing a steady expansion trend. However, animal-derived phospholipids have relatively lower production volumes due to their higher cost.

[0003] Chicken bone marrow is rich in nutrients, including fat, protein, vitamins, and various minerals, especially phospholipids, but it is often discarded as a byproduct. The various nutrients in chicken bone marrow help maintain nutritional balance, promote hemoglobin synthesis, bone development, and enhance immunity. In 2023, global chicken production was 103.418 million tons, and China's chicken production was 14.3 million tons, generating a large amount of waste chicken bone marrow annually. However, there is currently no industrial-scale extraction method for phospholipids from chicken bone marrow. Commonly used phospholipid extraction methods include organic solvent extraction, supercritical fluid extraction, compound salt precipitation, and column chromatography. Organic solvent extraction is a traditional method for extracting phospholipids. Its principle is to utilize the different solubilities of phospholipids and other components to be separated in different organic solvents to achieve separation. The key is finding suitable organic solvent extractants with good solubility and selectivity for phospholipids. Supercritical fluid extraction has disadvantages such as high equipment cost, the need to operate under high pressure, and limited sample throughput, making it unsuitable for large-scale production and having significant limitations. The key to compound salt precipitation lies in selecting an effective and suitable inorganic salt precipitant. Column chromatography has limitations such as long operation time, limited sample volume, and the use of some toxic organic solvents. Summary of the Invention

[0004] To address at least one of the aforementioned technical problems, this invention provides chicken bone marrow phospholipids, their preparation method, and applications, specifically employing the following technical solution:

[0005] In a first aspect, the present invention provides a method for extracting phospholipids from chicken bone marrow, comprising the following steps:

[0006] S1: Freeze-dry chicken bone marrow to obtain freeze-dried chicken bone marrow powder;

[0007] S2: Place the freeze-dried chicken bone marrow powder in a mixed solution of dichloromethane and methanol, mix well, and obtain a chicken bone marrow mixed solution;

[0008] S3: The chicken bone marrow mixture solution is subjected to ultra-fine pulverization by a high-energy fluid mill and then centrifuged to obtain the supernatant.

[0009] S4: Mix the upper clear liquid with water, stir, and let stand to obtain the lower layer of dichloromethane and the upper mixed liquid;

[0010] S5: The upper mixture is repeatedly extracted with dichloromethane 2-3 times to obtain dichloromethane mixtures;

[0011] S6: Combine the lower layer of dichloromethane described in S4 and the dichloromethane mixture described in S5, concentrate under reduced pressure, and dry to obtain mixed liposomes;

[0012] S7: Dissolve the mixed liposomes in acetone, extract, centrifuge, and dry to obtain the chicken bone marrow phospholipids.

[0013] As a further preferred embodiment, the volume ratio of the above-mentioned freeze-dried chicken bone marrow powder to the mixed solution of dichloromethane and methanol is 1:5 to 1:20 (g / L). More preferably, the volume ratio of the freeze-dried chicken bone marrow powder to the mixed solution of dichloromethane and methanol is 1:10. In the above preparation process, (1) an excessively high material-to-liquid ratio will result in excessive solvent, and due to uneven distribution and the dilution effect of the material in the solvent, the extraction efficiency will decrease. In addition, an excessively high material-to-liquid ratio may also make subsequent processing (such as solid-liquid separation, solvent recovery, etc.) difficult, increasing production costs; (2) an excessively low material-to-liquid ratio will result in the material becoming viscous, which will affect the penetration and diffusion of the solvent, further reducing the extraction efficiency. At the same time, viscous material may also clog the extraction equipment, affecting the continuity and stability of production.

[0014] As a further preferred embodiment, the volume ratio of dichloromethane to methanol is 2:1 to 4:1. Phospholipids are amphiphilic substances, and methanol has a relatively weak solubility for polar substances. In the above preparation process, (1) if the proportion of methanol is too high, it will lead to insufficient solubility for specific phospholipids, thereby affecting the extraction effect. (2) If the proportion of dichloromethane is too high, it may reduce the solubility for specific phospholipid components. The two need to reach the optimal ratio to maximize the phospholipid yield.

[0015] As a further preferred embodiment, the pressure of the high-energy fluid milling process is 30~120 MPa. More preferably, the pressure of the high-energy fluid milling process is 120 MPa. In the above preparation process, (1) if the high-energy fluid milling process pressure is too low, the cell wall breaking effect on chicken bone marrow cells will be poor, thereby reducing the phospholipid yield. (2) if the high-energy fluid milling process pressure is too high, the temperature will rise sharply, causing phospholipid oxidation, thereby reducing the nutritional value of phospholipids.

[0016] As a further preferred embodiment, the centrifugation speed is 4000 r / min and the centrifugation time is 30 min.

[0017] As a further preferred embodiment, the above-mentioned vacuum concentration is carried out using a rotary evaporator at a temperature of 25 °C.

[0018] As a further preferred embodiment, the drying described in S6 and S7 is performed at a constant temperature of 25°C.

[0019] Secondly, the present invention provides chicken bone marrow phospholipids prepared by the above preparation method.

[0020] Thirdly, the present invention provides the application of the above-mentioned chicken bone marrow phospholipids in the preparation of drugs for diseases related to free radicals.

[0021] The beneficial effects of this invention are as follows:

[0022] This invention is simple to operate and low in cost. It utilizes a lipid extraction method combined with high-energy fluid milling technology, employing low-toxicity dichloromethane instead of chloroform, resulting in a high yield of chicken bone marrow phospholipids. Compared to existing organic solvent extraction methods, this invention offers advantages such as higher extraction efficiency, shorter extraction time, and solvent savings. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0024] Figure 1 This is a graph showing the results of the determination of the antioxidant activity (DPPH) of chicken bone marrow phospholipids provided in some embodiments of this application;

[0025] Figure 2 This is a graph showing the results of the determination of the antioxidant activity (ABTS) of chicken bone marrow phospholipids provided in some embodiments of this application. Detailed Implementation

[0026] The technical solutions in the embodiments of this application will be clearly and completely described below with reference to the embodiments of this application. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0027] Example 1

[0028] A method for extracting phospholipids from chicken bone marrow, which specifically includes the following steps:

[0029] Step 1, Pretreatment Process - Freeze Drying

[0030] Pre-cool the freeze dryer for 30 minutes, then pre-cool it to -80°C. Take out the frozen chicken bone marrow, quickly place it on a tray into the freeze dryer, cover it with an acrylic tube, and press the vacuum switch to create a vacuum. Once the vacuum level of the freeze dryer reaches below 20 Pa, start the freeze drying process. During the freeze drying process, the vacuum level of the freeze dryer is always controlled below 20 Pa. After the chicken bone marrow is freeze-dried for 48 hours, it is removed from the freeze dryer to obtain freeze-dried chicken bone marrow powder (which is reddish-brown, loose in texture, slightly glossy, and has a fishy odor).

[0031] Step 2: Take 5 kg of freeze-dried chicken bone marrow powder into a container, add 25 L of dichloromethane-methanol (volume ratio 2:1) composite extract (material-liquid ratio 1:5). Prepare the extract fresh before use. Homogenize for 1 min to ensure thorough mixing. Process the mixture through a high-energy fluid mill at 120 MPa and centrifuge at 4000 r / min for 30 min. Take the supernatant.

[0032] Step 3: After mixing the supernatant and deionized water from Step 2, let it stand and collect the lower layer of dichloromethane. Add 10 L of dichloromethane to the remaining supernatant and solids for a second extraction. Repeat the above extraction operation twice. Combine the collected dichloromethane and transfer the combined dichloromethane into a vacuum evaporator. Concentrate under reduced pressure at 25 °C to obtain a chicken bone marrow lipid mixture. Dry the chicken bone marrow lipid mixture in a vacuum oven at a constant temperature of 25 °C to obtain a lipid mixture.

[0033] Step 4: After the lipid mixture from Step 3 has cooled, add 10 L of acetone, shake thoroughly to extract, centrifuge at 4000 r / min for 30 min, collect the precipitate and dry it at 25 ℃ in an oven to obtain chicken bone marrow phospholipids. Store the dried chicken bone marrow phospholipids in a refrigerator at 4 ℃. In this example, the yield of chicken bone marrow phospholipids was 0.98%.

[0034] Example 2

[0035] A method for extracting phospholipids from chicken bone marrow, which specifically includes the following steps:

[0036] Step 1, Pretreatment Process - Freeze Drying

[0037] Pre-cool the freeze dryer for 30 minutes, then pre-cool it to -80°C. Take out the frozen chicken bone marrow, quickly place it on a tray into the freeze dryer, cover it with an acrylic tube, and press the vacuum switch to create a vacuum. Once the vacuum level of the freeze dryer reaches below 20 Pa, start the freeze drying process. During the freeze drying process, the vacuum level of the freeze dryer is always controlled below 20 Pa. After the chicken bone marrow is freeze-dried for 48 hours, it is removed from the freeze dryer to obtain freeze-dried chicken bone marrow powder (which is reddish-brown, loose in texture, slightly glossy, and has a fishy odor).

[0038] Step 2: Take 5 kg of freeze-dried chicken bone marrow powder into a container, add 50 L of dichloromethane-methanol (volume ratio 2:1) compound extraction solution (material-liquid ratio 1:10). Prepare the extraction solution fresh for use. Homogenize for 1 min to ensure thorough mixing. Process the mixture through a high-energy fluid mill at 120 MPa and centrifuge at 4000 r / min for 30 min. Take the supernatant.

[0039] Step 3: After mixing the supernatant and deionized water from Step 2 thoroughly, let it stand and collect the lower layer of dichloromethane. Add 20 L of dichloromethane to the remaining supernatant and solids for a second extraction. Repeat the above extraction operation twice. Combine the collected dichloromethane and transfer the combined dichloromethane into a vacuum evaporator. Concentrate under reduced pressure at 25 °C to obtain a chicken bone marrow lipid mixture. Dry the chicken bone marrow lipid mixture in a vacuum oven at a constant temperature of 25 °C to obtain a lipid mixture.

[0040] Step 4: After the lipid mixture from Step 3 has cooled, add 10 L of acetone, shake thoroughly to extract, centrifuge at 4000 r / min for 30 min, collect the precipitate and dry it at 25 ℃ in an oven to obtain chicken bone marrow phospholipids. Store the dried chicken bone marrow phospholipids in a refrigerator at 4 ℃. In this example, the yield of chicken bone marrow phospholipids was 1.52%.

[0041] Example 3

[0042] A method for extracting phospholipids from chicken bone marrow, which specifically includes the following steps:

[0043] Step 1, Pretreatment Process - Freeze Drying

[0044] Pre-cool the freeze dryer for 30 minutes, then pre-cool it to -80°C. Take out the frozen chicken bone marrow, quickly place it on a tray into the freeze dryer, cover it with an acrylic tube, and press the vacuum switch to create a vacuum. Once the vacuum level of the freeze dryer reaches below 20 Pa, start the freeze drying process. During the freeze drying process, the vacuum level of the freeze dryer is always controlled below 20 Pa. After the chicken bone marrow is freeze-dried for 48 hours, it is removed from the freeze dryer to obtain freeze-dried chicken bone marrow powder (which is reddish-brown, loose in texture, slightly glossy, and has a fishy odor).

[0045] Step 2: Take 5 kg of freeze-dried chicken bone marrow powder into a container, add 100 L (material-liquid ratio 1:20) of dichloromethane-methanol (volume ratio 2:1) composite extract. Prepare the extract fresh before use. Homogenize for 1 min to ensure thorough mixing. Process the mixture through a high-energy fluid mill at 120 MPa and centrifuge at 4000 r / min for 30 min. Take the supernatant.

[0046] Step 3: After mixing the supernatant and deionized water from Step 2, let it stand and collect the lower layer of dichloromethane. Add 40 L of dichloromethane to the remaining supernatant and solids for a second extraction. Repeat the above extraction operation twice. Combine the collected dichloromethane and transfer the combined dichloromethane into a vacuum evaporator. Concentrate under reduced pressure at 25 °C to obtain a chicken bone marrow lipid mixture. Dry the chicken bone marrow lipid mixture in a vacuum oven at a constant temperature of 25 °C to obtain a lipid mixture.

[0047] Step 4: After the lipid mixture from Step 3 has cooled, add 10 L of acetone, shake thoroughly to extract, centrifuge at 4000 r / min for 30 min, collect the precipitate and dry it at 25 ℃ in an oven to obtain chicken bone marrow phospholipids. Store the dried chicken bone marrow phospholipids in a refrigerator at 4 ℃. In this example, the yield of chicken bone marrow phospholipids was 1.47%.

[0048] Example 4

[0049] A method for extracting phospholipids from chicken bone marrow, which specifically includes the following steps:

[0050] Step 1, Pretreatment Process - Freeze Drying

[0051] Pre-cool the freeze dryer for 30 minutes, then pre-cool it to -80°C. Take out the frozen chicken bone marrow, quickly place it on a tray into the freeze dryer, cover it with an acrylic tube, and press the vacuum switch to create a vacuum. Once the vacuum level of the freeze dryer reaches below 20 Pa, start the freeze drying process. During the freeze drying process, the vacuum level of the freeze dryer is always controlled below 20 Pa. After the chicken bone marrow is freeze-dried for 48 hours, it is removed from the freeze dryer to obtain freeze-dried chicken bone marrow powder (which is reddish-brown, loose in texture, slightly glossy, and has a fishy odor).

[0052] Step 2: Take 5 kg of freeze-dried chicken bone marrow powder into a container, add 25 L of dichloromethane-methanol (volume ratio 3:1) composite extract (material-liquid ratio 1:5). Prepare the extract fresh before use. Homogenize for 1 min to ensure thorough mixing. Process the mixture through a high-energy fluid mill at 120 MPa and centrifuge at 4000 r / min for 30 min. Take the supernatant.

[0053] Step 3: After mixing the supernatant and deionized water from Step 2, let it stand and collect the lower layer of dichloromethane. Add 10 L of dichloromethane to the remaining supernatant and solids for a second extraction. Repeat the above extraction operation twice. Combine the collected dichloromethane and transfer the combined dichloromethane into a vacuum evaporator. Concentrate under reduced pressure at 25 °C to obtain a chicken bone marrow lipid mixture. Dry the chicken bone marrow lipid mixture in a vacuum oven at a constant temperature of 25 °C to obtain a lipid mixture.

[0054] Step 4: After the lipid mixture from Step 3 has cooled, add 10 L of acetone, shake thoroughly to extract, centrifuge at 4000 r / min for 30 min, collect the precipitate and dry it at 25 ℃ in an oven to obtain chicken bone marrow phospholipids. Store the dried chicken bone marrow phospholipids in a refrigerator at 4 ℃. In this example, the yield of chicken bone marrow phospholipids is 1.05%.

[0055] Example 5

[0056] A method for extracting phospholipids from chicken bone marrow, which specifically includes the following steps:

[0057] Step 1, Pretreatment Process - Freeze Drying

[0058] Pre-cool the freeze dryer for 30 minutes, then pre-cool it to -80°C. Take out the frozen chicken bone marrow, quickly place it on a tray into the freeze dryer, cover it with an acrylic tube, and press the vacuum switch to create a vacuum. Once the vacuum level of the freeze dryer reaches below 20 Pa, start the freeze drying process. During the freeze drying process, the vacuum level of the freeze dryer is always controlled below 20 Pa. After the chicken bone marrow is freeze-dried for 48 hours, it is removed from the freeze dryer to obtain freeze-dried chicken bone marrow powder (which is reddish-brown, loose in texture, slightly glossy, and has a fishy odor).

[0059] Step 2: Take 5 kg of freeze-dried chicken bone marrow powder into a container, add 25 L of dichloromethane-methanol (volume ratio 4:1) composite extract (material-liquid ratio 1:5). Prepare the extract fresh before use. Homogenize for 1 min to ensure thorough mixing. Process the mixture through a high-energy fluid mill at 120 MPa and centrifuge at 4000 r / min for 30 min. Take the supernatant.

[0060] Step 3: After mixing the supernatant and deionized water from Step 2, let it stand and collect the lower layer of dichloromethane. Add 10 L of dichloromethane to the remaining supernatant and solids for a second extraction. Repeat the above extraction operation twice. Combine the collected dichloromethane and transfer the combined dichloromethane into a vacuum evaporator. Concentrate under reduced pressure at 25 °C to obtain a chicken bone marrow lipid mixture. Dry the chicken bone marrow lipid mixture in a vacuum oven at a constant temperature of 25 °C to obtain a lipid mixture.

[0061] Step 4: After the lipid mixture from Step 3 has cooled, add 10 L of acetone, shake thoroughly to extract, centrifuge at 4000 r / min for 30 min, collect the precipitate and dry it at 25 ℃ in an oven to obtain chicken bone marrow phospholipids. Store the dried chicken bone marrow phospholipids in a refrigerator at 4 ℃. In this example, the yield of chicken bone marrow phospholipids was 1.21%.

[0062] Comparative Example 1

[0063] A method for extracting phospholipids from chicken bone marrow, which specifically includes the following steps:

[0064] Step 1: Take 5 kg of fresh chicken bone marrow that has not been freeze-dried into a container, add 50 L of dichloromethane-methanol (volume ratio 2:1) compound extract (material-liquid ratio 1:10). Prepare the extract fresh before use. Homogenize for 1 min to mix thoroughly. Process with a high-energy fluid mill at 120 MPa and centrifuge at 4000 r / min for 30 min. Take the supernatant.

[0065] Step 2: After mixing the supernatant and deionized water from Step 2 thoroughly, let it stand and collect the lower layer of dichloromethane. Add 20 L of dichloromethane to the remaining supernatant and solids for a second extraction. Repeat the above extraction operation twice. Combine the collected dichloromethane and transfer the combined dichloromethane into a vacuum evaporator. Concentrate under reduced pressure at 25 °C to obtain a chicken bone marrow lipid mixture. Dry the chicken bone marrow lipid mixture in a vacuum oven at a constant temperature of 25 °C to obtain a lipid mixture.

[0066] Step 4: After the lipid mixture from Step 3 has cooled, add 10 L of acetone, shake thoroughly to extract, centrifuge at 4000 r / min for 30 min, collect the precipitate and dry it at 25 ℃ in an oven to obtain chicken bone marrow phospholipids. Store the dried chicken bone marrow phospholipids in a refrigerator at 4 ℃. In this example, the yield of chicken bone marrow phospholipids was 0.32%.

[0067] Comparative Example 2

[0068] A method for extracting phospholipids from chicken bone marrow, which specifically includes the following steps:

[0069] Step 1, Pretreatment Process - Freeze Drying

[0070] Pre-cool the freeze dryer for 30 minutes, then pre-cool it to -80°C. Take out the frozen chicken bone marrow, quickly place it on a tray into the freeze dryer, cover it with an acrylic tube, and press the vacuum switch to create a vacuum. Once the vacuum level of the freeze dryer reaches below 20 Pa, start the freeze drying process. During the freeze drying process, the vacuum level of the freeze dryer is always controlled below 20 Pa. After the chicken bone marrow is freeze-dried for 48 hours, it is removed from the freeze dryer to obtain freeze-dried chicken bone marrow powder (which is reddish-brown, loose in texture, slightly glossy, and has a fishy odor).

[0071] Step 2: Take 5 kg of freeze-dried chicken bone marrow powder into a container, add 50 L of dichloromethane-methanol (volume ratio 2:1) compound extraction solution (material-liquid ratio 1:10). Prepare the extraction solution fresh for use. Homogenize for 1 min to mix thoroughly, and centrifuge at 4000 r / min for 30 min. Take the supernatant.

[0072] Step 3: After mixing the supernatant and deionized water from Step 2 thoroughly, let it stand and collect the lower layer of dichloromethane. Add 20 L of dichloromethane to the remaining supernatant and solids for a second extraction. Repeat the above extraction operation twice. Combine the collected dichloromethane and transfer the combined dichloromethane into a vacuum evaporator. Concentrate under reduced pressure at 25 °C to obtain a chicken bone marrow lipid mixture. Dry the chicken bone marrow lipid mixture in a vacuum oven at a constant temperature of 25 °C to obtain a lipid mixture.

[0073] Step 4: After the lipid mixture from Step 3 has cooled, add 10 L of acetone, shake thoroughly to extract, centrifuge at 4000 r / min for 30 min, collect the precipitate and dry it at 25 ℃ in an oven to obtain chicken bone marrow phospholipids. Store the dried chicken bone marrow phospholipids in a refrigerator at 4 ℃. In this example, the yield of chicken bone marrow phospholipids was 0.41%.

[0074] The yield of chicken bone marrow phospholipids prepared in the embodiments of the present invention is calculated using the following formula (1);

[0075] Chicken bone marrow phospholipid extraction rate:

[0076] (1)

[0077] Wherein, W1 is the phospholipid yield (%), m0 is the mass of the freeze-dried chicken bone marrow powder (g), and m2 is the mass of the extracted chicken bone marrow phospholipids (g).

[0078] The results are shown in Table 1:

[0079] Table 1

[0080]

[0081] Example 6

[0082] Determination of antioxidant activity of chicken bone marrow phospholipids

[0083] The chicken bone phospholipids prepared according to this invention were compared with commercially available soybean phospholipids for oxidation or oxidative activity determination.

[0084] 1. DPPH free radical scavenging test method

[0085] Samples of different concentrations were mixed with an equal volume of 0.1 mmol / L DPPH ethanol solution. The mixture was allowed to react at room temperature in the dark for 30 min. The absorbance was measured at a wavelength of 515 nm. The formula for calculating the DPPH free radical scavenging effect is as follows:

[0086] DPPH free radical scavenging effect (%) = Free radical scavenging activity (%) = [1 - (A - A0) / A1] * 100%

[0087] In the formula, A0 is the absorbance value obtained by replacing the DPPH solution with an equal volume of ethanol, A1 is the absorbance value obtained by replacing the sample solution with an equal volume of distilled water, and A is the absorbance value obtained by the sample.

[0088] 2. ABTS Free Radical Scavenging Test Method

[0089] A mixture of 5 mL ABTS (7 mm) and 88 mL potassium persulfate (40 mm) was prepared and reacted in the dark at room temperature for 12 h to generate ABTS free radical cations. Before use, the absorbance of the prepared mixture was adjusted to 0.70 ± 0.02 with ethanol to obtain the ABTS working solution. Subsequently, 0.4 mL of the sample solution was mixed with 3.6 mL of the ABTS working solution and incubated in the dark at room temperature for 6 min. The absorbance value was measured at a wavelength of 734 nm. The formula for calculating the ABTS free radical scavenging activity is as follows:

[0090] ABTS free radical scavenging effect (%) = Free radical scavenging activity (%) = [1 - (B - B0) / B1] * 100%

[0091] In the formula, B0 is the absorbance value obtained by replacing the ABTS solution with an equal volume of ethanol, B1 is the absorbance value obtained by replacing the sample solution with an equal volume of distilled water, and B is the absorbance value obtained by the sample.

[0092] like Figure 1 and 2 As shown, the antioxidant capacity of chicken bone marrow phospholipids and commercially available soybean phospholipids was compared using DPPH and ABTS free radical scavenging assays. Chicken bone marrow phospholipids exhibited strong DPPH free radical scavenging activity at concentrations ranging from 3 to 12 mg / mL, while commercially available soybean phospholipids showed lower DPPH free radical scavenging activity. Similarly, in the ABTS assay, chicken bone marrow phospholipids effectively scavenged positively charged ABTS free radicals, while commercially available soybean phospholipids showed poor ABTS free radical scavenging effect. Therefore, the chicken bone marrow phospholipids obtained by this patented method have a better antioxidant effect than commercially available soybean phospholipids, which plays an important role in preventing many diseases caused by free radicals, such as common cancers, arteriosclerosis, and diabetes.

[0093] This document uses specific examples to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the core ideas of this application. However, this application is not limited to the specific implementation methods described above. The specific implementation methods described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of this application without departing from the spirit and scope of the claims, and all of these forms are within the protection scope of this application.

Claims

1. A method for extracting phospholipids from chicken bone marrow, characterized in that, Includes the following steps: S1: Freeze-dry chicken bone marrow to obtain freeze-dried chicken bone marrow powder; S2: Place the freeze-dried chicken bone marrow powder in a mixed solution of dichloromethane and methanol, mix well, and obtain a chicken bone marrow mixed solution; S3: The chicken bone marrow mixture solution is subjected to ultra-fine pulverization by a high-energy fluid mill and then centrifuged to obtain the supernatant. S4: Mix the upper clear liquid with water, stir, and let stand to obtain the lower layer of dichloromethane and the upper mixed liquid; S5: The upper mixture is repeatedly extracted with dichloromethane 2-3 times to obtain dichloromethane mixtures; S6: Combine the lower layer of dichloromethane described in S4 and the dichloromethane mixture described in S5, concentrate under reduced pressure, and dry to obtain mixed liposomes; S7: Dissolve the mixed liposomes in acetone, extract, centrifuge, and dry to obtain the chicken bone marrow phospholipids; The mass-to-volume ratio of the freeze-dried chicken bone marrow powder to the mixed solution of dichloromethane and methanol is 1g:5~20L; The volume ratio of the dichloromethane to the methanol is 2:1 to 4:1; The pressure of the high-energy fluid mill treatment is 30 MPa to 120 MPa.

2. The extraction method according to claim 1, characterized in that, The freeze-drying temperature is -75 ℃ to -85 ℃, and the vacuum degree is less than 20 Pa.

3. The extraction method according to claim 1, characterized in that, The centrifugation speed was 4000 r / min, and the centrifugation time was 30 min.

4. The extraction method according to claim 1, characterized in that, The vacuum concentration was carried out using a rotary evaporator at a temperature of 25 °C.

5. The extraction method according to claim 1, characterized in that, The drying described in S6 and S7 is performed at a constant temperature of 25°C.

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