Earthworm peptide composition and application thereof in reducing blood lipid

Abstract

The application discloses a kind of earthworm peptide composition and its application in lowering blood fat, belong to functional food technical field.The earthworm peptide composition provided by the application is composed of earthworm peptide mixture and hawthorn extract, and the hawthorn extract and earthworm polypeptide mixture are mutually synergized in function, and the effect is superimposed, good blood fat lowering, health maintenance effect is achieved;And long-acting, high safety;The preparation method of the earthworm peptide composition provided, mild conditions, no organic toxic reagent is used, and no toxic and harmful substance is generated, and it is environment-friendly;The preparation method is clear, and can be mass-produced.

Description

Technical Field

[0001] This invention relates to the field of health food technology, and in particular to an earthworm peptide composition and its application in lowering blood lipids. Background Technology

[0002] Blood lipids mainly include cholesterol (total cholesterol, low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C)) and triglycerides, with cholesterol being the most important component in blood lipid tests. Excess LDL-C in the blood can slowly deposit on the walls of blood vessels, forming atherosclerotic plaques. This leads to narrowing and blockage of blood vessels, causing symptoms such as chest tightness, dizziness, and intermittent claudication, potentially affecting the heart (coronary artery disease), brain (insufficient blood supply to the brain), kidneys, or lower limbs. Unstable plaques can detach and directly block blood vessels, forming thrombi and leading to acute myocardial infarction or cerebral infarction. When triglycerides in the blood are severely elevated, a large number of chylomicrons appear, increasing blood viscosity and making it easier to block pancreatic microvessels, potentially triggering acute pancreatitis. Severe pancreatitis has a high mortality rate. HDL-C, on the other hand, helps clear LDL-C from blood vessels, reducing the risk of cardiovascular disease.

[0003] High intake of saturated fats (including red meat, fatty meat, organ meats, butter, full-fat dairy products, and palm oil found in many processed foods), high intake of trans fats (fried foods, pastries, biscuits, margarine, non-dairy creamer, etc.), high sugar, and alcohol are the main factors contributing to secondary hyperlipidemia. In addition, insufficient dietary fiber intake, lack of exercise (prolonged sitting lowers HDL-C levels), and smoking (damages the vascular endothelium and also lowers HDL-C) are also unhealthy lifestyle habits that can lead to elevated blood lipids.

[0004] Therefore, developing health foods that can lower blood lipids for daily dietary intervention to prevent hyperlipidemia and related cardiovascular and cerebrovascular diseases from the source is of great practical significance for safely and continuously improving the health level of the whole people and reducing the medical burden. Summary of the Invention

[0005] The purpose of this invention is to provide an earthworm peptide composition and its application in lowering blood lipids, providing a blood lipid-lowering health food with good blood lipid-lowering effect, simple composition, and good safety for promoting health through dietary therapy.

[0006] To achieve the above objectives, the present invention provides an earthworm peptide composition, wherein the composition comprises an earthworm peptide mixture and a hawthorn extract; the mass ratio of the earthworm peptide mixture preparation raw material to the hawthorn extract preparation raw material is 1:1. The earthworm peptide mixture consists of product I and product II, and is prepared by the following method: S1. After crushing fresh or freeze-dried earthworms, sonicate to obtain a homogenate. Add buffer solution to the homogenate to adjust the pH. After stirring and reacting, centrifuge to collect the supernatant. After purification, freeze-dry and record as product I. S2. Take the precipitate obtained after centrifugation in S1, add anhydrous ethanol, stir the reaction, centrifuge to take the supernatant, and rotary evaporate to recover the anhydrous ethanol. The liquid obtained is denoted as product II. In S1, the ultrasound is conducted at 35-40 kHz for 5-10 minutes; the buffer solution is phosphate buffer, and the pH is adjusted to 7.5-8.0 using hydrochloric acid / sodium hydroxide. The hawthorn extract consists of extract I and extract II, and the preparation steps are as follows: (1) Take fresh or dried hawthorn, chop / crush it and sieve it. Add it to an ethanol-water solution, reflux the reaction, take the supernatant and evaporate it to recover the ethanol. After concentration, extract I is obtained. (2) Add the precipitate after taking the supernatant from (1) to drinking water, boil it, keep it warm, and concentrate it to obtain extract II; The ethanol aqueous solution in (1) is a 70% ethanol aqueous solution by volume, and the amount added is 2-3 times the mass of fresh hawthorn and 5-6 times the mass of dried hawthorn; the reflux reaction is refluxed at 60℃ for 3-4 hours.

[0007] Preferably, in step S1, fresh earthworms are minced and then added to 1-2 times their volume of drinking water before being ultrasonicated; dried earthworms are ground into powder and then added to 5-6 times their volume of drinking water before being ultrasonicated. The temperature is kept not higher than 40-50°C during both the grinding and ultrasonication processes.

[0008] Preferably, the stirring reaction in S1 is carried out at 4°C and 180-200 rpm for 48-72 hours; purification is carried out by ammonium sulfate precipitation, and the precipitate is collected and freeze-dried.

[0009] Preferably, the amount of anhydrous ethanol added in S2 is 1-2 times the precipitate volume; the stirring reaction is carried out at 40°C and 180-200 rpm for 60-90 minutes.

[0010] Preferably, in step (2), the heat preservation time is 60-90 minutes, and the concentration is carried out by rotary evaporation.

[0011] The earthworm peptide composition described above is used in the preparation of health food products, which help maintain healthy blood lipid levels; the health food products use the earthworm peptide composition as a functional component.

[0012] Earthworm (Pheretima aspergillum) is a traditional Chinese medicine, specifically the dried body of several species of earthworms (Pheretima spp., Pheretima simonii, Pheretima wieldii, or Pheretima protozoa) belonging to the family Lumbriaceae. Traditional Chinese medicine theory considers earthworm to be cold in nature and salty in taste, primarily affecting the liver, spleen, and bladder meridians. Its main functions, as stated in the *Chinese Pharmacopoeia* and related TCM classics, are: clearing heat and calming the nerves, promoting blood circulation and removing blood stasis, clearing the lungs and relieving asthma, and clearing heat and promoting diuresis.

[0013] Earthworms contain active enzymes such as lumbrokinase and plasmin, which can directly degrade fibrin in thrombi and activate the body's fibrinolytic system, working synergistically to exert highly effective anticoagulant and thrombolytic effects.

[0014] Earthworms are rich in unsaturated fatty acids, which are important components for regulating blood lipids and preventing cardiovascular and cerebrovascular diseases. They can also steadily lower blood pressure by regulating the relaxation of lipid smooth muscle and enhance the body's antioxidant capacity, thereby delaying atherosclerosis.

[0015] Earthworms are also rich in protein and amino acids, with the content in dried products reaching 50%-70%. They contain 18 kinds of amino acids, including the 8 essential amino acids, making them a high-quality source of complete animal protein. They are also rich in iron, calcium, phosphorus, selenium, and other trace elements beneficial to the human body, as well as abundant B vitamins and vitamin D.

[0016] Therefore, the earthworm peptide composition provided by the present invention and its application in lowering blood lipids have the following specific technical effects: (1) The total flavonoids (such as hyperoside and quercetin), triterpenoid acids (ursolic acid and oleanolic acid), pectin, and phytosterols in hawthorn can reduce the synthesis of cholesterol and the body's absorption of cholesterol by inhibiting HMG-CoA reductase and ACAT activity. They can also promote the decomposition and metabolism of cholesterol and the clearance of bad cholesterol by increasing CYP7A1 enzyme activity, increasing LDL-C receptor levels, and activating the PPAR-α pathway. The soluble dietary fiber (such as pectin) in the extract can directly bind to cholesterol, further reducing the body's absorption of cholesterol, while promoting intestinal peristalsis and improving the excretion capacity / efficiency of cholesterol. Hawthorn extract plays a role in "treating the root cause" in traditional Chinese medicine, mainly by reducing cholesterol production at the source.

[0017] (2) The lumbrokinase, plasmin, highly unsaturated fatty acids, polypeptides and free amino acids in the earthworm polypeptide mixture can, on the one hand, directly degrade fibrin and dissolve plaques and thrombi on the inner wall of blood vessels; on the other hand, they can activate the body's fibrinolytic system and enhance the body's ability to dissolve plaques / LDL-C on the inner wall of blood vessels; in addition, the highly unsaturated fatty acids in it can also regulate blood lipids and resist atherosclerosis, thereby improving blood rheology and reducing blood viscosity.

[0018] The mixture of hawthorn extract and earthworm polypeptide has synergistic effects and additive results, achieving good effects in lowering blood lipids and maintaining health.

[0019] (3) The earthworm peptide composition provided by the present invention has the effect of lowering blood lipids, and at the same time, the action time is long and the safety is high; the preparation method of the earthworm peptide composition provided is mild, does not use organic toxic reagents, and does not generate toxic and harmful substances, which is environmentally friendly; the preparation method is clear and can be mass-produced.

[0020] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0021] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments of the present invention 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.

[0022] Figure 1 These are photographs of zebrafish from each treatment group in the efficacy testing section of this invention; where A is an example image of the zebrafish tail blood vessel triglyceride staining intensity analysis area; B is the normal control group; C is the model control group; D is the positive control group; and E is the earthworm peptide group. Figure 2 These are the quantitative results of Oil Red O staining in each treatment group in the efficacy test section of this invention; where *** indicates the difference compared to the model control group. P <0.001. Detailed Implementation

[0023] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0024] To make the objectives, technical solutions, and advantages of this application clearer, more thorough, and more complete, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and embodiments. The following detailed descriptions are all illustrations of embodiments, intended to provide further detailed explanation of the present invention. Unless otherwise specified, all technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0025] The instruments, equipment, reagents and materials used in the embodiments were all obtained through commercial means; the methods and steps not described in detail in the embodiments are all conventional techniques in the art.

[0026] Example 1 The specific steps for preparing a mixture of earthworm peptides are as follows: (1) Rinse the purchased live earthworms with running drinking water to remove soil and feed residue. After draining the water, weigh one portion (203g), mince it, add 200g of drinking water, sonicate at 35KHz for 5 minutes, sonicate for 30 seconds and cool on ice for 5 minutes. The total sonication time is 5 minutes, and ensure that the temperature during the sonication process does not exceed 40℃.

[0027] (2) Add 1 volume of phosphate buffer to the homogenate obtained in step (1), stir and mix well, then adjust the pH to 7.8 with hydrochloric acid / sodium hydroxide, and stir at 4℃ and 200 rpm for 48 hours. Then centrifuge at 10000 rpm for 10 minutes and take the supernatant (the precipitate is recorded as precipitate I and stored in a refrigerator at 4℃ for later use) into an Erlenmeyer flask, and add 80% ammonium sulfate solution dropwise until the precipitate no longer increases. After each addition, gently shake the Erlenmeyer flask to observe the precipitation situation and decide whether to add it again.

[0028] After the addition was complete, centrifuge at 10,000 rpm for 10 minutes and collect the precipitate (referred to as precipitate II). Freeze-dry under vacuum to obtain product I, totaling 41.6 g, and store at 4°C for later use.

[0029] (3) Add 1 volume of anhydrous ethanol to the precipitate I obtained in step (2), stir and react for 60 minutes at 40°C and 200 rpm, centrifuge at 10000 rpm for 10 minutes to take the supernatant, and rotary evaporate the anhydrous ethanol. The resulting pale yellow to yellow clear oily liquid is product II.

[0030] Example 2 The specific steps for preparing a mixture of earthworm peptides are as follows: (1) Grind the purchased freeze-dried earthworm Chinese medicine into powder (after adding liquid nitrogen to grind the powder) through a 60-mesh sieve, weigh 200g, grind it, add 1200g of drinking water, sonicate at 35KHz for 5 minutes, sonicate for 30 seconds and cool on ice for 5 minutes, the total sonication time is 5 minutes, and ensure that the temperature during the sonication process does not exceed 40℃.

[0031] (2) Add 1 volume of phosphate buffer (pH 7.8) to the homogenate obtained in step (1) and stir at 4°C and 200 rpm for 48 hours. Then centrifuge at 10,000 rpm for 10 minutes and take the supernatant (the precipitate is called precipitate I and stored at 4°C for later use) into an Erlenmeyer flask. Add 80% ammonium sulfate solution dropwise until the precipitate no longer increases. After each addition, gently shake the Erlenmeyer flask to observe the precipitation and decide whether to add more solution.

[0032] After the addition was complete, centrifuge at 10,000 rpm for 10 minutes and collect the precipitate (referred to as precipitate II). Freeze-dry under vacuum to obtain product I, and store at 4°C for later use.

[0033] (3) Add 1 volume of anhydrous ethanol to the precipitate I obtained in step (2), stir and react for 60 minutes at 40°C and 200 rpm, centrifuge at 10000 rpm for 10 minutes to take the supernatant, and rotary evaporate the anhydrous ethanol. The resulting pale yellow to yellow clear oily liquid is product II.

[0034] Example 3 The specific steps for preparing a hawthorn extract are as follows: (1) After removing rotten and moldy fruit from the purchased fresh hawthorn, rinse it with running water, then clean the surface with a soft brush and rinse it again with running drinking water. After removing the pit, persistent calyx and calyx tube, chop it (weigh a total of 209g), add 400g of 70% (v / v) ethanol aqueous solution, reflux at 60℃ for 3 hours, centrifuge and take the supernatant, recover the ethanol by rotary evaporation, concentrate to 100mL, and record it as extract I.

[0035] (2) Add the precipitate obtained in step (1) to 2 times the volume of drinking water, heat to 100°C and keep it at a gentle boil for 60 minutes, then concentrate by rotary evaporation to 150 mL, which is recorded as extract II.

[0036] Example 4 The specific steps for preparing a hawthorn extract are as follows: (1) Remove the pits from the purchased dried hawthorn slices, crush them through a 60-mesh sieve, weigh 200g and add 1000g of 70% (v / v) ethanol aqueous solution, reflux at 60℃ for 3 hours, centrifuge and take the supernatant, recover the ethanol by rotary evaporation and concentrate to 100mL, which is recorded as extract I.

[0037] (2) Add the precipitate obtained in step (1) to 2 times the volume of drinking water, heat to 100°C and keep it at a gentle boil for 60 minutes, then concentrate by rotary evaporation to 150 mL, which is recorded as extract II.

[0038] Example 5 The specific steps for preparing a hawthorn extract are as follows: (1) After removing rotten and moldy fruit from the purchased fresh hawthorn, rinse it with running water, then clean the surface with a soft brush and rinse it again with running drinking water. After removing the pit, persistent calyx and calyx tube, chop it (weigh a total of 209g), add 400mL of 70% (v / v) ethanol aqueous solution, reflux at 80℃ for 3 hours, centrifuge and take the supernatant, recover the ethanol by rotary evaporation, concentrate to 100mL, and record it as extract I.

[0039] (2) Add the precipitate obtained in step (1) to 2 times the volume of drinking water, heat to 100°C and keep it at a gentle boil for 60 minutes, then concentrate by rotary evaporation to 150 mL, which is recorded as extract II.

[0040] Example 6 Prepare an earthworm peptide composition, and the specific steps are as follows: Mix the Product I and Product II prepared in Example 1, and Extract I and Extract II prepared in Example 3, then filter through a 0.22 μm filter membrane in a laminar flow hood, seal in a brown glass bottle, and store at 4°C.

[0041] Effect test All zebrafish used in the experiment were raised in fish culture water at 28°C (water quality: add 200 mg of instant sea salt to every 1 L of reverse osmosis water, conductivity is 450 - 550 μS / cm; pH is 6.5 - 8.5; hardness is 50 - 100 mg / L CaCO3). The license number for the use of experimental animals is: SYXK(Zhe)2022 - 0004, and the feeding management meets the requirements of international AAALAC accreditation (accreditation number: 001458). The IACUC ethical review number is: IACUC - 2025 - 202511110008 - 01.

[0042] (1) Determination of the maximum tolerated concentration (MTC).

[0043] Randomly select wild - type AB - strain zebrafish at 5 days post - fertilization (5dpf) into beakers. Add 25 mL of fish culture water + 30 zebrafish to each beaker, and set up a total of 7 beakers: Randomly select 1 beaker, and the zebrafish in it are not treated, denoted as the normal control group; Randomly select 1 beaker, and the zebrafish in the beaker are given a high - sugar and high - fat diet in water solution, denoted as the model control group; Randomly select 1 beaker, and the zebrafish in the beaker are given a high - sugar and high - fat diet in water solution + 125 μg / mL of the earthworm peptide composition prepared in Example 6, denoted as the earthworm peptide group 1; Randomly select 1 beaker, and the zebrafish in the beaker are given a high - sugar and high - fat diet in water solution + 250 μg / mL of the earthworm peptide composition prepared in Example 6, denoted as the earthworm peptide group 2; Randomly select 1 beaker, and the zebrafish in the beaker are given a high - sugar and high - fat diet in water solution + 500 μg / mL of the earthworm peptide composition prepared in Example 6, denoted as the earthworm peptide group 3; Randomly select 1 beaker, and the zebrafish in the beaker are given a high - sugar and high - fat diet in water solution + 1000 μg / mL of the earthworm peptide composition prepared in Example 6, denoted as the earthworm peptide group 4; For the remaining 1 beaker, the zebrafish in the beaker are given a high - sugar and high - fat diet in water solution + 2000 μg / mL of the earthworm peptide composition prepared in Example 6, denoted as the earthworm peptide group 5. Except for the normal control group, the other experimental groups are all given the special high - sugar and high - fat diet for zebrafish in water solution. The addition amount of the earthworm peptide composition is calculated based on the polypeptide content in the measured composition.

[0044] After zebrafish in each treatment group were treated at 28℃ for 2 days, the phenotype of zebrafish in each treatment group was observed on the third day, the number of dead zebrafish was counted, and the mortality rate was calculated (mortality rate = number of dead zebrafish / 30 × 100%). The statistical results are shown in Table 1. The earthworm peptide composition prepared in Example 6 was added to 2000 μg / mL and no obvious toxic effect was observed.

[0045] Table 1. Results of MTC investigation of the earthworm peptide composition prepared in Example 1 on model zebrafish.

[0046] (2) Evaluation of lipid-lowering efficacy.

[0047] Wild-type AB strain zebrafish, 5 days post-fertilization (5 dpf), were randomly selected and placed in beakers. Each beaker contained 25 mL of aquarium water and 30 zebrafish, for a total of four beakers: one beaker containing untreated zebrafish (normal control); one beaker containing zebrafish given a water-soluble high-sugar, high-fat diet (model control); one beaker containing zebrafish given a water-soluble high-sugar, high-fat diet plus 2000 μg / mL of the earthworm peptide composition prepared in Example 6 (earthworm peptide group); and the remaining beaker containing zebrafish given a water-soluble high-sugar, high-fat diet plus 11.6 μg / mL of atorvastatin calcium (positive control). The high-sugar, high-fat diet added to each treatment group (excluding the normal control group) was a zebrafish-specific high-sugar, high-fat diet. The amount of earthworm peptide composition added was based on the determined polypeptide content of the composition.

[0048] Zebrafish from each treatment group were treated at 28℃ for 2 days, and then immediately collected for whole-body fat staining with Oil Red O. After staining, 10 zebrafish from each experimental group were randomly selected and photographed under a dissecting microscope. The images were saved, and data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The intensity of triglyceride staining in the blood vessels of the zebrafish tail was analyzed and statistically analyzed. The lipid-lowering efficacy of the samples was evaluated based on the statistical analysis results of this index. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS software. P <0.05 indicates that the difference is statistically significant. Results are as follows... Figures 1-2 As shown in Table 2, where Figure 1 In the diagram, A represents an example of the area analyzed for the staining intensity of triglycerides in the blood vessels of zebrafish tails; B represents the normal control group; C represents the model control group; D represents the positive control group; and E represents the earthworm peptide group. Figure 2 The staining intensity of the title on the vertical axis represents the staining intensity of triglycerides in the blood vessels of the zebrafish tail, in pixels.

[0049] Table 2. Results of the experiment evaluating the lipid-lowering efficacy.

[0050] Note: *** indicates a comparison with the model control group. P <0.001.

[0051] The prepared earthworm peptide composition uses earthworm (a traditional Chinese medicinal herb) and hawthorn (a food and medicine homology) as raw materials. All reagents used in the preparation process are highly safe or have low toxicity, making it suitable for use in or preparation of health foods. The zebrafish experiments described above also demonstrate that the prepared earthworm peptide composition is highly safe for consumption; no significant toxic effects were observed when the composition was added to drinking water at a concentration of 2000 μg / mL.

[0052] Therefore, the earthworm peptide composition provided by this invention consists of an earthworm peptide mixture and hawthorn extract. The hawthorn extract and the earthworm peptide mixture have synergistic functions and additive effects, achieving good effects in lowering blood lipids and maintaining health. Moreover, the effect is long-lasting and highly safe. The preparation method of the earthworm peptide composition provided is mild, does not use organic toxic reagents, and does not generate any toxic or harmful substances, making it environmentally friendly. The preparation method is clear and can be mass-produced.

[0053] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A composition of earthworm peptides, characterized in that: The composition helps maintain healthy blood lipid levels; the composition consists of a mixture of earthworm peptides and hawthorn extract; the mass ratio of the raw materials for preparing the earthworm peptide mixture to the raw materials for preparing the hawthorn extract is 1:

1. The earthworm peptide mixture consists of product I and product II, and is prepared by the following method: S1. After crushing fresh or freeze-dried earthworms, sonicate to obtain a homogenate. Add buffer solution to the homogenate to adjust the pH. After stirring and reacting, centrifuge to collect the supernatant. After purification, freeze-dry and record as product I. S2. Take the precipitate obtained after centrifugation in S1, add anhydrous ethanol, stir the reaction, centrifuge to take the supernatant, and rotary evaporate to recover the anhydrous ethanol. The liquid obtained is denoted as product II. In S1, the ultrasound is conducted at 35-40 kHz for 5-10 minutes; the buffer solution is phosphate buffer, and the pH is adjusted to 7.5-8.0 using hydrochloric acid / sodium hydroxide. The hawthorn extract consists of extract I and extract II, and the preparation steps are as follows: (1) Take fresh or dried hawthorn, chop / crush it and sieve it. Add it to an ethanol-water solution, reflux the reaction, take the supernatant and evaporate it to recover the ethanol. After concentration, extract I is obtained. (2) Add the precipitate after taking the supernatant from (1) to drinking water, boil it, keep it warm, and concentrate it to obtain extract II; The ethanol aqueous solution in (1) is a 70% ethanol aqueous solution by volume, and the amount added is 2-3 times the mass of fresh hawthorn and 5-6 times the mass of dried hawthorn; the reflux reaction is refluxed at 60℃ for 3-4 hours.

2. The earthworm peptide composition according to claim 1, characterized in that: In step S1, fresh earthworms are minced and then mixed with 1-2 times the volume of drinking water before being ultrasonicated. Dried earthworms are ground into powder and then mixed with 5-6 times the volume of drinking water before being ultrasonicated. The temperature is kept below 40-50℃ during both the grinding and ultrasonication processes.

3. The earthworm peptide composition according to claim 1, characterized in that: The stirring reaction in S1 is carried out at 4°C and 180-200 rpm for 48-72 hours; purification is carried out by ammonium sulfate precipitation, and the precipitate is collected and freeze-dried.

4. The earthworm peptide composition according to claim 1, characterized in that: The amount of anhydrous ethanol added in S2 is 1-2 times the volume of the precipitate; the stirring reaction is carried out at 40°C and 180-200 rpm for 60-90 minutes.

5. The earthworm peptide composition according to claim 1, characterized in that: In step (2), the temperature is maintained for 60-90 minutes, and the concentration is achieved by rotary evaporation.

6. The application of the earthworm peptide composition according to any one of claims 1-5 in the preparation of health food, characterized in that: The health food helps maintain healthy blood lipid levels; the health food uses earthworm peptide composition as a functional component.

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