A high-purity phospholipid extraction and enrichment method based on antarctic krill oil

Abstract

The application provides a high-purity phospholipid extraction and enrichment method based on Antarctic krill oil, and belongs to the technical field of phospholipid purification. Specifically, high-purity phospholipid extraction is realized based on marine phospholipid directional enrichment technology, a food-grade ethanol-water-glycerol extraction system is used, glycerol reduces PC precipitation and loss through hydrogen bonds, and the aggregation of phospholipid phases and the reduction of viscosity can be prevented in the concentration stage; food-grade activated carbon in the system avoids agglomeration with the help of hydrogen bonds with glycerol and adsorbs impurities; ascorbic acid palmitate and citric acid are added as stabilizers, and the biological activity of phospholipids is preserved by combining low temperature and inert gas protection; neutral alumina is used instead of silica gel to separate PC and PE for purification, and the purity of PC after treatment is high.

Description

Technical Field

[0001] This invention belongs to the field of phospholipid purification technology, specifically, it relates to a method for extracting and enriching high-purity phospholipids based on Antarctic krill oil. Background Technology

[0002] Phospholipids are an important raw material for functional foods and high-end health products, and their purity, especially the purity of phosphatidylcholine (PC), directly determines the quality of the products.

[0003] Currently, traditional phospholipid extraction processes mostly employ a single alcohol-water extraction system. Polyphosphate (PC) is prone to precipitation and aggregation, and it easily binds non-specifically to adsorbents. During the concentration stage, phospholipid phase aggregation and high system viscosity are common. Adsorbents also tend to agglomerate within the system, resulting in low impurity removal efficiency. Phospholipids are susceptible to oxidative degradation during processing, leading to reduced bioactivity. Furthermore, the purification process often uses silica gel as the stationary phase in column chromatography, which lacks sufficient selective enrichment of the target phosphatidylcholine component. These numerous problems with traditional processes result in inconsistent quality of the final phospholipid products, failing to consistently meet the stringent standards for functional foods, high-end health supplement raw materials, pharmaceutical excipients, and active pharmaceutical ingredients, thus limiting the application of phospholipids in high-end fields.

[0004] Therefore, developing a marine phospholipid targeted enrichment technology that can solve the above-mentioned technical pain points has become an urgent technical need to be addressed in this field. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention uses a food-grade ethanol-water-glycerol extraction system. The three hydroxyl groups of glycerol can form hydrogen bonds with the phosphate groups of PC, reducing PC precipitation and aggregation, as well as non-specific binding with the adsorbent to minimize losses. Its high boiling point and low vapor pressure also prevent phospholipid aggregation and reduce the viscosity of the concentrate during the concentration stage, ensuring smooth subsequent operations. Food-grade activated carbon introduced into the system forms hydrogen bonds with glycerol through its surface groups, preventing aggregation and efficiently adsorbing impurities such as free fatty acids. Simultaneously, food-grade ascorbate palmitate and citric acid are added throughout the process as stabilizers. Citric acid and glycerol form a weakly acidic microenvironment, extending the stabilization time, while ascorbate palmitate forms an antioxidant protective layer on the surface of phospholipid micelles to inhibit phospholipid oxidation. Low temperatures and inert gas protection throughout the process preserve the bioactivity of phospholipids. Neutral alumina replaces silica gel in the purification stage, adsorbing PE and separating PC through specific coordination bonds. The resulting PC has high purity, meeting the standards for functional foods, high-end health product raw materials, pharmaceutical excipients, and active pharmaceutical ingredients.

[0006] To achieve the above objectives, the technical solution adopted by this invention is a method for extracting and enriching high-purity phospholipids based on Antarctic krill oil, the specific process of which is as follows: S1: Weigh 100 parts of Antarctic krill oil with a content of 40%, introduce nitrogen gas with a flow rate of 50 mL / min into it, add 0.2 parts of citric acid, heat at 45℃ for 20 min, stir at 1500 r / min, and cool naturally to room temperature to obtain the complex precursor. S2: Weigh 2% by mass of medium-chain triglycerides and add them to the complex precursor obtained in step S1. Heat at 45°C for 20 min and stir at 800 r / min to obtain the ternary extract. S3: Weigh 85-100 parts of ethanol, 15-25 parts of water and 3-5 parts of glycerol and mix them at a speed of 500 r / min. Weigh 0.05-0.1 parts of ascorbate palmitate and 6 parts of activated carbon and add them to the mixture. Sonicate at a frequency of 25 kHz for 15 min and continue stirring at the speed of 500 r / min to obtain the extract. S4: The ternary extract obtained in step S2 is added dropwise to the extract obtained in step S3, stirred at 500 r / min, heated at 30℃ for 30 min, then allowed to stand at -15℃ for 6-8 h, centrifuged at 10000 r / min for 20 min, the upper extract is collected and concentrated to 80% at 45℃ to obtain crude phospholipid solution. S5: Weigh 10 parts of neutral alumina and dry it at 110℃ for 4 hours. After cooling to room temperature, add the alumina to 4-6 parts of ethanol and stir for 20 minutes at 800 r / min to obtain a paste. Pack the paste into a 30 mm × 300 mm chromatography column, add 45 parts of ethanol and pass it through the column (this operation is repeated twice). Manually pressurize the column, and close the stopcock when the liquid level is 1 cm from the top surface of the packing. Let it stand for 30 minutes to allow the sample to fully combine with the stationary phase, obtaining the packed chromatography column. Add the crude phospholipid solution obtained in step S4 to the packed chromatography column and elute stepwise: Phase 1: Use 50 portions of 85% ethanol containing 0.2% glycerol, with an elution flow rate of 3 ml / min; Second stage: Use 50 portions of 90% ethanol containing 0.1% glycerol, with an elution flow rate of 3 ml / min; Third stage: Use 50 portions of 95% ethanol to regenerate and elute the chromatography column at a flow rate of 3 ml / min.

[0007] The eluent was collected and subjected to reduced pressure treatment at 45℃ and -0.2MPa for 2 hours, and then naturally cooled to room temperature to obtain high-purity phospholipids.

[0008] The beneficial effects achieved by this invention are as follows: This invention discloses a high-purity phospholipid extraction and enrichment method using a food-grade ethanol-water-glycerol extraction system. The glycerol molecule contains three hydroxyl groups, which can form hydrogen bonds with the phosphate groups of phosphatidylcholine (PC), effectively reducing the precipitation and aggregation of PC in the ethanol-water system, improving its dispersion stability at low temperatures, reducing non-specific binding between PC and the adsorbent, and minimizing the loss of the target product. Simultaneously, glycerol's high boiling point and low vapor pressure allow it to act as a plasticizer during the concentration stage, preventing excessive water loss and aggregation of the phospholipid phase, as well as structural densification. It also reduces the viscosity of the concentrate, improves system fluidity, and ensures smooth subsequent operations. The food-grade activated carbon introduced into the system has surface carboxyl and hydroxyl groups that interact with the hydroxyl groups of glycerol through hydrogen bonds, forming steric hindrance. This effectively prevents activated carbon agglomeration, maintains good dispersion, and increases the effective adsorption area, thereby efficiently adsorbing free fatty acids, phospholipid oxidation products, and pro-oxidative metal ions in the system, achieving impurity removal.

[0009] The high-purity phospholipid extraction and enrichment method of this invention, to further improve process stability and PC product quality, incorporates food-grade ascorbyl palmitate and citric acid as natural stabilizers throughout the process: citric acid and glycerol interact through hydrogen bonds to form a weakly acidic microenvironment, reducing adsorption loss of citric acid at the solid-liquid interface and prolonging its stabilizing time; ascorbyl palmitate is uniformly distributed at the oil-water interface, with its lipophilic portion facing the lipid phase and its hydrophilic ascorbic acid portion facing the aqueous phase, forming a dense interfacial antioxidant protective layer on the surface of phospholipid micelles, effectively inhibiting phospholipid oxidative degradation. Regarding the protection of active ingredients, a low-temperature operation mode is used throughout the process, coupled with inert gas protection, to minimize oxidative degradation of phospholipids during processing and preserve their biological activity. In the purification process, neutral alumina is used instead of traditional silica gel as the stationary phase in column chromatography. The empty orbitals of aluminum atoms on the alumina surface serve as Lewis acid sites, forming specific coordination bonds with the lone pair electrons of the amino group in phosphatidylethanolamine. Under low-polarity elution conditions, PE can be firmly adsorbed, while PC, lacking strong Lewis basic groups, cannot form stable coordination and is thus effectively separated from the eluent. After processing with this marine phospholipid targeted enrichment technology, the purity of PC is superior to that of conventional polar adsorption separation methods such as silica gel column chromatography. Attached Figure Description

[0010] Figure 1 This is a process flow diagram of the high-purity phospholipid extraction and enrichment method proposed in this application; Figure 2 The high-purity phospholipid product obtained in Example 2; Figure 3 The HPLC test results are shown for the high-purity phospholipids after separation and purification in Example 2 and Comparative Example 3. Figure 4The graphs show the test results for phospholipid purity and phospholipid recovery rate obtained in the examples and comparative examples. Figure 5 The graphs show the test results for MDA content in the high-purity phospholipids obtained in the examples and comparative examples. Figure 6 The graph shows the test results of the removal rate of phosphatidylethanolamine in the high-purity phospholipids obtained in the examples and comparative examples. Figure 7 The images show test results for molds and yeasts in high-purity phospholipids obtained in the examples and comparative examples.

[0011] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof. Detailed Implementation

[0012] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0013] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. The preferred embodiments and materials described herein are for illustrative purposes only and do not limit the scope of this application.

[0014] The extraction and enrichment methods in the following examples and comparative examples refer to Figure 1 physical reference Figure 2 The test results are referenced. Figures 3-7 Unless otherwise specified, all methods are conventional. Unless otherwise specified, all materials used in the following examples are new materials purchased from the market and are food grade. All parts mentioned in the instructions are parts by weight, and the medium-chain triglycerides are triglycerides of decanoic acid (C10).

[0015] Example 1: A method for extracting and enriching high-purity phospholipids based on Antarctic krill oil, the specific process of which is as follows: S1: Weigh 100 parts of Antarctic krill oil with a content of 40%, introduce nitrogen gas with a flow rate of 50 mL / min into it, add 0.2 parts of citric acid, heat at 45℃ for 20 min, stir at 1500 r / min, and cool naturally to room temperature to obtain the complex precursor. S2: Weigh 2% by mass of medium-chain triglycerides and add them to the complex precursor obtained in step S1. Heat at 45°C for 20 min and stir at 800 r / min to obtain the ternary extract. S3: Weigh 85 parts ethanol, 15 parts water and 3 parts glycerol and mix them at a speed of 500 r / min. Weigh 0.05 parts ascorbate palmitate and 6 parts activated carbon and add them to the mixture. Sonicate at a frequency of 25 kHz for 15 min and continue stirring at the speed mentioned above for 30 min to obtain the extract. S4: The ternary extract obtained in step S2 is added dropwise to the extract obtained in step S3, stirred at 500 r / min, heated at 30℃ for 30 min, then allowed to stand at -15℃ for 6 h, centrifuged at 10000 r / min for 20 min, the upper extract is collected and concentrated to 80% at 45℃ to obtain crude phospholipid solution. S5: Weigh 10 parts of neutral alumina and dry it at 110℃ for 4 hours. After cooling to room temperature, add the alumina to 4 parts of ethanol and stir for 20 minutes at 800 r / min to obtain a paste. Pack the paste into a 30 mm × 300 mm chromatography column, add 45 parts of ethanol and pass it through the column (this operation is repeated twice). Manually pressurize the column, and close the stopcock when the liquid level is 1 cm from the top surface of the packing. Let it stand for 30 minutes to allow the sample to fully combine with the stationary phase, obtaining the packed chromatography column. Add the crude phospholipid solution obtained in step S4 to the packed chromatography column and elute stepwise: Phase 1: Use 50 portions of 85% ethanol containing 0.2% glycerol, with an elution flow rate of 3 ml / min; Second stage: Use 50 portions of 90% ethanol containing 0.1% glycerol, with an elution flow rate of 3 ml / min; Third stage: Use 50 portions of 95% ethanol to regenerate and elute the chromatography column at a flow rate of 3 ml / min.

[0016] The eluent was collected and subjected to reduced pressure treatment at 45℃ and -0.2MPa for 2 hours, and then naturally cooled to room temperature to obtain high-purity phospholipids.

[0017] Example 2: A method for extracting and enriching high-purity phospholipids based on Antarctic krill oil, the specific process of which is as follows: S1: Weigh 100 parts of Antarctic krill oil with a content of 40%, introduce nitrogen gas with a flow rate of 50 mL / min into it, add 0.2 parts of citric acid, heat at 45℃ for 20 min, stir at 1500 r / min, and cool naturally to room temperature to obtain the complex precursor. S2: Weigh 2% by mass of medium-chain triglycerides and add them to the complex precursor obtained in step S1. Heat at 45°C for 20 min and stir at 800 r / min to obtain the ternary extract. S3: Weigh 95 parts of ethanol, 20 parts of water and 4 parts of glycerol and mix them at a speed of 500 r / min. Weigh 0.08 parts of ascorbate palmitate and 6 parts of activated carbon and add them to the mixture. Sonicate at a frequency of 25 kHz for 15 min and continue stirring at the speed mentioned above for 30 min to obtain the extract. S4: The ternary extract obtained in step S2 is added dropwise to the extract obtained in step S3, stirred at 500 r / min, heated at 30℃ for 30 min, then allowed to stand at -15℃ for 7 h, centrifuged at 10000 r / min for 20 min, the upper extract is collected and concentrated to 80% at 45℃ to obtain crude phospholipid solution. S5: Weigh 10 parts of neutral alumina and filter it through a 1μm filter membrane. Dry it at 110℃ for 4 hours. After cooling to room temperature, add the alumina to 5 parts of ethanol and stir for 20 minutes at 800 r / min to obtain a paste. Pack the paste into a 30mm×300mm chromatography column, add 45 parts of ethanol and pass it through the column (this operation is repeated twice). Manually pressurize the column. When the liquid level is 1cm from the top surface of the packing material, close the stopcock and let it stand for 30 minutes to allow the sample to fully combine with the stationary phase, obtaining a packed chromatography column. Add the crude phospholipid solution obtained in step S4 to the packed chromatography column and elute stepwise: Phase 1: Use 50 portions of 85% ethanol containing 0.2% glycerol, with an elution flow rate of 3 ml / min; Second stage: Use 50 portions of 90% ethanol containing 0.1% glycerol, with an elution flow rate of 3 ml / min; Third stage: Use 50 portions of 95% ethanol to regenerate and elute the chromatography column at a flow rate of 3 ml / min.

[0018] The eluent was collected and subjected to reduced pressure treatment at 45℃ and -0.2MPa for 2 hours, and then naturally cooled to room temperature to obtain high-purity phospholipids.

[0019] Example 3: A method for extracting and enriching high-purity phospholipids based on Antarctic krill oil, the specific process of which is as follows: S1: Weigh 100 parts of Antarctic krill oil with a content of 40%, introduce nitrogen gas with a flow rate of 50 mL / min into it, add 0.2 parts of citric acid, heat at 45℃ for 20 min, stir at 1500 r / min, and cool naturally to room temperature to obtain the complex precursor. S2: Weigh 2% by mass of medium-chain triglycerides and add them to the complex precursor obtained in step S1. Heat at 45°C for 20 min and stir at 800 r / min to obtain the ternary extract. S3: Weigh 100 parts of ethanol, 25 parts of water and 5 parts of glycerol and mix them at a speed of 500 r / min. Weigh 0.1 parts of ascorbate palmitate and 6 parts of activated carbon and add them to the mixture. Sonicate at a frequency of 25 kHz for 15 min and continue stirring at the speed mentioned above for 30 min to obtain the extract. S4: The ternary extract obtained in step S2 is added dropwise to the extract obtained in step S3, stirred at 500 r / min, heated at 30℃ for 30 min, then allowed to stand at -15℃ for 8 h, centrifuged at 10000 r / min for 20 min, the upper extract is collected and concentrated to 80% at 45℃ to obtain crude phospholipid solution. S5: Weigh 10 parts of neutral alumina and filter it through a 1μm filter membrane. Dry it at 110℃ for 4 hours. After cooling to room temperature, add the alumina to 6 parts of ethanol and stir for 20 minutes at 800 r / min to obtain a paste. Pack the paste into a 30mm×300mm chromatography column, add 45 parts of ethanol and pass it through the column (this operation is repeated twice). Manually pressurize the column. When the liquid level is 1cm from the top surface of the packing material, close the stopcock and let it stand for 30 minutes to allow the sample to fully combine with the stationary phase, obtaining a packed chromatography column. Add the crude phospholipid solution obtained in step S4 to the packed chromatography column and elute stepwise: Phase 1: Use 50 portions of 85% ethanol containing 0.2% glycerol, with an elution flow rate of 3 ml / min; Second stage: Use 50 portions of 90% ethanol containing 0.1% glycerol, with an elution flow rate of 3 ml / min; Third stage: Use 50 portions of 95% ethanol to regenerate and elute the chromatography column at a flow rate of 3 ml / min.

[0020] The eluent was collected and subjected to reduced pressure treatment at 45℃ and -0.2MPa for 2 hours, and then naturally cooled to room temperature to obtain high-purity phospholipids.

[0021] Comparative example: The difference between Comparative Example 1 and Example 2 is that the latter was not processed in step S5, while the rest is the same as Example 2; The difference between Comparative Example 2 and Example 2 is that the extract obtained in step S3 was not added; the rest of the extract is the same as in Example 2. The difference between Comparative Example 3 and Example 2 is that the Antarctic krill oil is only processed in step S5.

[0022] The following tests were performed on the high-purity phospholipids obtained through extraction and enrichment: Figure 3 The results of HPLC detection of high-purity phospholipids obtained in Example 2 and Comparative Example 3 (mobile phase: acetonitrile-ethanol-water (volume ratio 30:63:7), flow rate: 1.0 mL / min, injection volume: 20 μL, column temperature: 25℃) show that the detection residence time of phospholipids is 9.75 min. The detection intensity in Example 2 is significantly higher than that in Comparative Example 3. Figure 4 This document compares the purity and recovery rate of the high-purity phospholipid products obtained in the examples and comparative examples. The results show that the phosphatidylcholine purity of Example 2 reaches 99.5%, significantly higher than the product obtained by the comparative process. Simultaneously, the recovery rate of the target product in Example 2 is 43.7%, an increase of approximately 23.4% compared to 35.4% in Comparative Example 2. These data demonstrate that the neutral alumina column chromatography process of this invention can significantly improve the recovery efficiency of the target component while ensuring high purity, solving the problem of difficulty in simultaneously achieving purity and recovery rate in traditional processes. Figure 5 The graph shows the changes in malondialdehyde (MDA) content of high-purity phospholipids obtained in the examples and comparative examples under accelerated storage conditions (specific detection procedure: thiobarbituric acid (TBA) colorimetric method was used. 0.1 g of high-purity phospholipid sample was weighed, 5 mL of 10% trichloroacetic acid solution was added, and the mixture was ultrasonically extracted for 10 min, centrifuged at 4000 r / min for 10 min; 2 mL of supernatant was taken, 2 mL of 0.67% TBA solution was added, and the mixture was reacted in a boiling water bath for 20 min. After cooling, the absorbance was measured at a wavelength of 532 nm. Accelerated aging of the high-purity phospholipids was achieved by heating at 60℃, and different sampling times were set: 0, 3, 6, 9, 12, and 15 days). The results show that with the extension of storage time, the MDA content of the example samples increased slowly and remained at a low level; while the MDA content of the comparative example samples increased significantly over time. Since MDA is a characteristic secondary product of phospholipid oxidative degradation, its content change directly reflects the oxidative stability of the sample. The above results demonstrate that the high-purity phospholipids prepared by the process of this invention exhibit significantly better oxidative stability than the comparative example. Figure 6 The results of phosphatidylethanolamine (PE) removal rate tests for the examples and comparative examples are shown. Example 2 achieved a PE removal rate of 100%, significantly higher than the 71.3% of Comparative Example 3, indicating that the process of this invention can efficiently remove PE impurities and achieve high-purity enrichment of PC. Figure 7 The results of mold, yeast, and total bacterial count tests on the high-purity phospholipids obtained in the examples and comparative examples are shown. The total bacterial count of the raw material, Antarctic krill oil, was 215 CFU / g. The microbial indicators of the phospholipid samples obtained in the examples were significantly lower than those of the raw crude oil and the comparative samples. This indicates that the extraction and purification process of the present invention can effectively reduce microbial contamination, and the resulting high-purity phospholipids have excellent hygiene indicators, demonstrating good biosafety and food safety compliance.

[0023] Table 1 shows the physicochemical properties of the high-purity phospholipids obtained in the examples and comparative examples:

[0024] Table 1 shows the physicochemical properties of the high-purity phospholipids obtained in the examples and comparative examples. As can be seen from the table, heavy metals (lead, arsenic, mercury, copper, and iron) were not detected in the products obtained in the examples of this invention; the moisture content was less than 0.4%; the acid value was less than 0.2 mg KOH / g; the iodine value remained between 123–127 g I2 / 100g; and the product color was pale yellow, transparent, and clear. Compared with the comparative example, the products of the examples had lower moisture content, lower acid value, higher iodine value, and better clarity.

[0025] The above results show that the high-purity phospholipid products prepared by the process of the present invention have excellent physicochemical properties, no risk of heavy metal pollution, and the moisture and acid values ​​are controlled at extremely low levels, which can effectively delay hydrolysis and oxidation during storage. The iodine value is maintained at a high level, indicating that the unsaturated fatty acid chains of the phospholipids are intact and have good biological activity.

[0026] Obviously, the above comparative examples and embodiments are only a part of the comparative examples and embodiments of the present invention, and they, along with the comparative examples and embodiments referenced based on such examples, are all within the scope of protection of this invention.

[0027] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

[0028] The present invention and its embodiments have been described above. This description is not restrictive, and the accompanying drawings are only one embodiment of the present invention. The actual application is not limited to this. In conclusion, if those skilled in the art are inspired by this description and design similar methods and embodiments without departing from the spirit of the present invention, they should all fall within the protection scope of the present invention.

Claims

1. A method for extracting and enriching high-purity phospholipids based on Antarctic krill oil, characterized in that, The enrichment method Includes the following processes: S1: Antarctic krill oil was pretreated, aerated, and citric acid was added to prepare a complex precursor; S2: Medium-chain triglycerides are added to the complex precursor, heated, and stirred to obtain a ternary extract; S3: Weigh ethanol, water and glycerol, stir, weigh ascorbate palmitate and activated charcoal, sonicate, stir, and obtain the extract; S4: Add the ternary extract to the extract, stir, heat, let stand at low temperature, centrifuge, collect the upper extract and concentrate to obtain crude phospholipid solution; S5: Pre-treat the chromatography column to obtain a packed chromatography column. Add the crude phospholipid solution to the packed chromatography column, elute stepwise, collect the eluent, reduce pressure, and obtain high-purity phospholipid.

2. The method for high-purity phospholipid extraction and enrichment based on Antarctic krill oil according to claim 1, characterized in that, The gas used for ventilation in step S1 is nitrogen, the flow rate of the nitrogen is 50 mL / min, the mass ratio of citric acid to Antarctic krill oil is 0.2:100, the heating temperature is 45°C, and the heating time is 20 min.

3. The method for high-purity phospholipid extraction and enrichment based on Antarctic krill oil according to claim 1, characterized in that, The medium-chain triglyceride in step S2 accounts for 2% of the mass of the complex precursor.

4. The method for high-purity phospholipid extraction and enrichment based on Antarctic krill oil according to claim 1, characterized in that, In step S3, the mass ratio of ethanol, water, glycerol, ascorbate palmitate, and activated carbon is 85-100:15-25:3-5:0.05-0.1:6, the frequency of the ultrasound is 25 kHz, and the duration of the ultrasound is 15 min.

5. The method for high-purity phospholipid extraction and enrichment based on Antarctic krill oil according to claim 1, characterized in that, The heating temperature in step S4 is 30°C, the heating time is 30 min, the low-temperature settling temperature is -15°C, and the low-temperature settling time is 6-8 h.

6. The method for high-purity phospholipid extraction and enrichment based on Antarctic krill oil according to claim 1, characterized in that, The pretreatment process of the chromatography column described in step S5 is as follows: Dry neutral alumina, add it to ethanol, stir to obtain a paste, pack the paste into a chromatography column, add ethanol, and obtain a packed chromatography column.

7. The method for high-purity phospholipid extraction and enrichment based on Antarctic krill oil according to claim 1, characterized in that, The stepwise elution process described in step S5 is as follows: Phase 1: Use 85% ethanol containing 0.2% glycerol; Second stage: Use 90% ethanol containing 0.1% glycerol; The third stage: The chromatography column is regenerated and eluted using 95% ethanol.

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