A green processing technology of sea cucumber powder with directional enrichment and improved polysaccharide release rate

Abstract

The application discloses a kind of green processing technology of sea cucumber powder with directional enrichment and improved polysaccharide release rate, and belongs to the field of food processing.The sea cucumber powder is prepared by the following steps: S1, sea cucumber pretreatment;S2, sea cucumber nutrient enrichment;S3, sea cucumber powder particle size control;S4, "single heavy" supercritical;S5, in situ ultrafine physical crushing;S6, "double heavy" supercritical.The application designs a gas distributor and adds a second hole disc design, so that the supercritical extraction instrument can effectively extract supermicro sea cucumber powder with supercritical carbon dioxide, achieving the deodorization of supermicro sea cucumber powder and improving the food quality of supermicro sea cucumber powder.Furthermore, through the detection and control of bulk density and tap density, the optimal processing parameters for supermicro sea cucumber powder are found, so that the prepared supermicro sea cucumber powder has high polysaccharide release rate, good food quality and easy subsequent processing.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of directional enrichment, promote the green processing technology of sea cucumber powder polysaccharide release rate, belong to food processing field. BACKGROUND

[0002] Sea cucumber (Stichopus japonicus) belongs to the echinoderm, sea cucumber class, benthic marine organism, is a kind of valuable marine biological resources. Sea cucumber is known for its high protein, low fat nutritional characteristics, also contains rich trace elements and various amino acids. Among them, sea cucumber polysaccharide is one of the important active ingredients of sea cucumber body wall, and its content accounts for 4% to 10% of the total organic matter of dry sea cucumber. Sea cucumber polysaccharide is an acidic mucopolysaccharide, which has strong biological activity in anticoagulation, hypolipidemic, immune regulation and anti-aging. However, sea cucumber polysaccharide is combined with protein through glycopeptide bond and hydrogen bond, so the absorption and utilization of polysaccharide requires the degradation of the connected protein.

[0003] In the prior art, in order to realize the effective separation of the two, the prior art proposes the concept of ultra-micro sea cucumber powder, such as CN 106509694A - a kind of edible convenient and easy-to-digest ultra-micro sea cucumber powder preparation, which discloses that the digestibility of the ultra-micro sea cucumber powder is improved, and the retention rate of crude polysaccharide content is high, which is 95.42%; and CN 105495411 A - a preparation method of sea cucumber ultra-micro powder sheet, which discloses that the sea cucumber ultra-micro powder of 1-30 μm reaches the specification of cell crushing, so that the active substances of sea cucumber are completely released. However, the processing technology of the ultra-micro sea cucumber powder in the foregoing two patents is relatively simple, and after simple washing, sterilization and enzyme inactivation, the sea cucumber powder is directly subjected to ultra-micro grinding, lacking a deodorization step, and a large number of odor molecules remain in the product, and the particle size of the ground sea cucumber powder is not measured by a target. Therefore, the final product has poor food quality, poor product purity and poor product quality.

[0004] Other methods for realizing the separation of protein and polysaccharide include hydrolysis with protease and extraction with alkaline solution, such as the method disclosed in patent CN 110204626A - a device and method for separating and refining sea cucumber polysaccharide. However, the use of alkaline solution can damage the structure of sea cucumber polysaccharide, causing the shedding of sugar groups, leading to the change of polysaccharide structure, so the concentration of alkaline solution and the reaction conditions need to be strictly controlled. Enzymatic hydrolysis requires control of enzyme concentration, enzymatic hydrolysis temperature and time, and this method has complicated operating conditions, and the enzymatic hydrolysis product has an unpleasant flavor, affecting the taste. Therefore, it is better to use enzymatic hydrolysis than direct physical grinding.

[0005] Therefore, the extraction and utilization of sea cucumber polysaccharide still need to be further explored to obtain a sea cucumber polysaccharide product with good food quality and excellent product quality, and a more excellent and effective method is used. SUMMARY

[0006] TECHNICAL PROBLEM

[0007] Sea cucumber polysaccharide has high use value, and it is necessary to separate sea cucumber polysaccharide from protein to utilize sea cucumber polysaccharide. Common separation methods include physical crushing method and enzymatic extraction method. The physical crushing method is simpler than the enzymatic extraction method, and the product quality is easier to control. However, the physical crushing method in the prior art is still simple, and the quality of the prepared sea cucumber powder is poor, the product purity is not good, and the quality is not easy to control. Therefore, it is necessary to improve it.

[0008] TECHNICAL CONTENT

[0009] The purpose of the present application is to overcome the shortcomings of the prior art sea cucumber polysaccharide and protein separation technology, and to provide a preparation method of sea cucumber powder with high polysaccharide release rate. The present application improves the sea cucumber polysaccharide release rate by using suitable physical crushing technology, and prepares a sea cucumber powder with high polysaccharide release rate and anti-atherosclerosis, and verifies it by using simulated digestion and animal model.

[0010] In order to achieve the above purpose, the present application provides a preparation method of sea cucumber powder with high polysaccharide release rate, which is prepared by the following steps:

[0011] S1, sea cucumber pretreatment: opening the abdomen of sea cucumber, removing the contents in the body cavity of sea cucumber, and washing the body wall of sea cucumber with water to obtain pretreated sea cucumber;

[0012] S2, sea cucumber nutrient enrichment: placing the pretreated sea cucumber obtained in step S1 in warm water for temporary cultivation, and then drying to obtain sea cucumber dry product;

[0013] S3, sea cucumber powder particle size control: crushing the sea cucumber dry product obtained in step S2 into sea cucumber coarse powder with a particle size of 5-30;

[0014] S4, "single" supercritical: placing the sea cucumber coarse powder obtained in step S3 in the extraction kettle of a supercritical extraction instrument for supercritical carbon dioxide extraction to remove odor;

[0015] S5, in-situ ultra-fine physical crushing: crushing the sea cucumber powder deodorized in step S4 by ultra-fine physical crushing to obtain a powder, and controlling the bulk density of the powder to be 0.65-0.70 g / mL, and the tap density to be 0.70-0.80 g / mL;

[0016] S6, "double" supercritical: placing the sea cucumber powder obtained in step S5 in the extraction kettle of a supercritical extraction instrument for supercritical carbon dioxide extraction to remove odor, and obtaining sea cucumber powder with high polysaccharide release rate.

[0017] Further, the temperature of the warm water in step S2 is 50-60℃.

[0018] Further, the time of the temporary cultivation in step S2 is 4-8h.

[0019] Further, the warm water temporary cultivation in step S2 is as follows: placing the pretreated sea cucumber in 50-60℃ warm water for 4-8h, and replacing the warm water every 0.5-1h.

[0020] Further, the drying in step S2 includes freeze drying, heating drying or air drying.

[0021] Further, the supercritical extraction instrument in step S4 and step S6 is a supercritical extraction instrument equipped with a gas distributor.

[0022] Further, the gas distributor in the supercritical extraction instrument is an improved gas distributor.

[0023] Further, the improved gas distributor comprises a mounting tube (1), a connecting flange (2), a first hole disc (3), a filter screen (4), a support column (5), a baffle (6), a telescopic rod (7), a spring (70), a pressing plate (8) and a second hole disc (9).

[0024] The mounting tube (1) is made of transparent material, facilitating observation of the internal situation. The upper and lower outer sides of the mounting tube (1) are connected with the connecting flanges (2), facilitating external connection of pipelines. The inner sides of the middle and lower parts of the mounting tube (1) are connected with the first hole discs (3). The lower sides of the first hole discs (3) are connected with the filter screens (4). Four support columns (5) are connected between the first hole discs (3). The upper sides of the support columns (5) are connected with the baffles (6), which are in contact with the mounting tube (1). The telescopic rods (7) are connected with the pressing plates (8) rotatably. The fixed ends and telescopic ends of the telescopic rods (7) are connected with the springs (70). The second hole disc (9) is placed between the baffles (6). The pressing plates (8) are in contact with the second hole disc (9). The first hole disc (3) and the second hole disc (9) have the same shape. Twelve conical parts are arranged on the upper side of the second hole disc (9), facilitating backflow.

[0025] Further, the parameters of the extraction kettle in step S4 are set as follows: temperature 30-40℃, extraction pressure 25-30MPa, CO2 flow rate 25-30L / h, and extraction time 2-2.5h.

[0026] Further, the precooling is performed before the pulverization in step S5 using a supermicro pulverizer. The precooling is performed using dry ice or liquid nitrogen, or by placing in a frozen environment. The temperature of the frozen environment is-80 to-20℃.

[0027] Further, the super-fine powder in step S5 is crushed under the rotation speed of 8000-12000 rpm and the mesh size of 0.3-0.6 mm.

[0028] Further, the bulk density in step S5 is 0.67-0.70 g / mL, and the tap density is 0.77-0.80 g / mL.

[0029] Specifically, the bulk density in step S5 is 0.70 g / mL, and the tap density is 0.80 g / mL.

[0030] Further, the parameters of the extraction kettle in step S6 are set as 30-40 ℃, the extraction pressure is 25-30 MPa, the CO2 flow rate is 25-30 L / h, and the extraction time is 2-2.5 h.

[0031] The application provides the sea cucumber powder with high polysaccharide release rate prepared according to the above method.

[0032] The application provides the sea cucumber powder with high polysaccharide release rate in the field of food, health care products or medicine preparation.

[0033] Further, the application in the field of health care products is to prepare health care products that can help maintain the healthy level of blood lipids (cholesterol / triglyceride).

[0034] Further, the application in the field of medicine preparation is to prepare medicines for preventing and treating atherosclerotic diseases.

[0035] The application has the following beneficial effects:

[0036] 1. The physical crushing method for separating the polysaccharide protein of sea cucumber in the prior art is relatively simple and lacks a deodorization step. The application can realize the deodorization of the super-fine sea cucumber powder by designing a gas distributor and installing a second hole disc, so as to improve the edible quality of the super-fine sea cucumber powder. The design of the second hole disc can also make the carbon dioxide fluid backflow to realize secondary extraction, thereby improving the extraction efficiency, effectively removing the odor molecules in the super-fine sea cucumber powder, and improving the purity of the sea cucumber polysaccharide in the product.

[0037] 2. The processing technology of the super-fine sea cucumber powder lacks parameter measurement and quality control. The application can find the optimal processing parameters for the super-fine sea cucumber powder by detecting and controlling the bulk density and tap density, so as to obtain the super-fine sea cucumber powder with high polysaccharide release rate, good edible quality and easy subsequent processing. In addition, the super-fine sea cucumber powder has excellent effects of resisting atherosclerosis and maintaining the healthy level of blood lipids, and is a product with wide market application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 Figure 1 is a perspective view of the improved gas disperser; wherein 1 is the mounting pipe, 2 is the connecting flange, 5 is the support column, 7 is the telescopic rod, 8 is the pressing plate, 9 is the second hole disc;

[0039] Figure 2 Figure 2 is a partial perspective view of the improved gas disperser; wherein 1 is the mounting pipe, 2 is the connecting flange, 3 is the first hole disc, 4 is the filter screen, 5 is the support column, 6 is the baffle, 7 is the telescopic rod, 8 is the pressing plate, 9 is the second hole disc;

[0040] Figure 3 Figure 3 is an enlarged perspective view of A of the improved gas disperser; wherein 1 is the mounting pipe, 5 is the support column, 6 is the baffle, 7 is the telescopic rod, 70 is the spring, 8 is the pressing plate, 9 is the second hole disc;

[0041] Figure 4 Figure 4 is a supercritical extraction device installed with the improved gas disperser, wherein the red arrow is the installation position of the improved gas disperser;

[0042] Figure 5 Figure 5 is the sea cucumber polysaccharide release curve of Example 1, 2 and Comparative Example 1;

[0043] Figure 6 Figure 6 is the blood cholesterol content of Example 1 and Comparative Example 1 after intervention on high-fat fed mice (different letters indicate significance);

[0044] Figure 7 Figure 7 is the aortic section of Example 1 and Comparative Example 1 after intervention on high-fat fed mice;

[0045] Figure 8 Figure 8 is the vascular wall thickness of Example 1 and Comparative Example 1 after intervention on high-fat fed mice (different letters indicate significance);

[0046] Figure 9 Figure 9 is the GC-IMS two-dimensional difference topography of Example 1;

[0047] Figure 10 Figure 10 is the GC-IMS two-dimensional difference topography of Comparative Example 3;

[0048] Figure 11 Figure 11 is the GC-IMS two-dimensional difference topography of Comparative Example 4. DETAILED DESCRIPTION

[0049] With reference to the drawings of the embodiments of the present application, the technical solutions in the embodiments of the present application will be clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments of the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative effort should fall within the protection scope of the present application.

[0050] In the present application, unless specifically defined and limited otherwise, the terms "connected", "connected", "fixed" should be understood broadly, for example, it can be fixed connection, or detachable connection, or integrated; it can be mechanical connection, or electrical connection; it can be directly connected, or indirectly connected through an intermediate medium, or the internal communication of two elements or the interaction relationship between two elements. For a person of ordinary skill in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0051] In the present application, unless specifically defined and limited otherwise, the first feature "on" or "under" the second feature can include that the first and second features are in direct contact, or that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, the first feature "on", "above" and "above" the second feature includes that the first feature is directly above and obliquely above the second feature, or only indicates that the horizontal height of the first feature is higher than that of the second feature. The first feature "under", "below" and "below" the second feature includes that the first feature is directly below and obliquely below the second feature, or only indicates that the horizontal height of the first feature is less than that of the second feature.

[0052] Experimental apparatus

[0053] The supercritical extraction instrument with improved gas distributor used in the examples and comparative examples is produced by Jiangsu Gaokeli Pharmaceutical Equipment Co., Ltd., and the model is GKSCFE220-40-30L. Among them, the gas distributor of the supercritical extraction instrument is improved, and the improved gas distributor is shown in Figures 1-3 .

[0054] The improved gas distributor includes a mounting pipe 1, a connecting flange 2, a first hole disc 3, a filter screen 4, a support column 5, a baffle 6, a telescopic rod 7, a spring 70, a pressing plate 8 and a second hole disc 9.

[0055] The installation pipe 1 is transparent, facilitating observation of the internal condition. The upper and lower portions of the installation pipe 1 are connected with connecting flanges 2, facilitating connection with external pipelines. The middle and lower inner sides of the installation pipe 1 are connected with first hole plates 3. The lower sides of the first hole plates 3 are connected with filter screens 4. Four support columns 5 are connected between the first hole plates 3. The upper sides of the support columns 5 are connected with baffles 6, which are in contact with the installation pipe 1. The baffles 6 are connected with telescopic rods 7. The fixed ends and telescopic ends of the telescopic rods 7 are connected with springs 70. The telescopic ends of the telescopic rods 7 are rotatably connected with pressing plates 8. The second hole plates 9 are placed between the baffles 6. The pressing plates 8 are in contact with the second hole plates 9. The first hole plates 3 and the second hole plates 9 are identical in shape. The upper side of the second hole plate 9 is provided with twelve conical parts, facilitating backflow.

[0056] When supercritical carbon dioxide extraction is performed, the improved gas distributor is used to drive the place where the extracted ingredients are required. The connecting flange 2 at the lower portion of the installation pipe 1 is connected with the feeding pipeline. The installation pipe 1 is transparent, facilitating observation of the internal condition. Then, the pressing plate 8 is pulled to move and rotate, so that the telescopic rod 7 is elongated, and the spring 70 is stretched. Then, the second hole plate 9 is brought into contact with the baffle 6. Then, the pressing plate 8 is reversely rotated to reset. Then, the pressing plate 8 is loosened. The spring 70 rebounds, and the telescopic rod 7 is contracted to restore, so that the pressing plate 8 is in contact with the second hole plate 9. Thus, the second hole plate 9 can be fixed, preventing the second hole plate 9 from falling due to the flow of carbon dioxide fluid. Then, the upper connecting flange 2 is connected with the feeding port of the extraction tank. The carbon dioxide fluid is added into the feeding pipeline, so that the carbon dioxide fluid enters the installation pipe 1, flows through the first hole plate 3, the filter screen 4 and the second hole plate 9, and then flows into the extraction tank to extract the material. The first hole plate 3 and the second hole plate 9 are identical in shape, so that the carbon dioxide fluid can be uniformly fed, improving the extraction efficiency and production efficiency. Meanwhile, the device can be placed in the extraction tank. The carbon dioxide can achieve the backflow effect under the action of the conical parts, so that the carbon dioxide fluid backflows under the action of the conical parts on the second hole plate 9, so that secondary extraction is performed, facilitating secondary extraction of the material. The pressure of the carbon dioxide fluid when discharged from the material basket is reduced, the material is prevented from being taken out, and the extraction effect is improved.

[0057] Example 1

[0058] S1, sea cucumber pretreatment: fresh sea cucumber is opened at the abdomen, the contents in the sea cucumber body cavity are removed, and the sea cucumber body wall is washed with water;

[0059] S2, sea cucumber nutrient enrichment: the pretreated sea cucumber obtained in step S1 is placed in warm water at 60℃ for temporary cultivation for 6h, the water is changed every 0.5h to remove the salt in the sea cucumber, and then the sea cucumber is fished out to remove the water by a vacuum freeze-drying machine for 24h to obtain sea cucumber dry product;

[0060] S3, sea cucumber powder particle size regulation: sea cucumber dry products are crushed into 20 mesh sea cucumber coarse powder in a pulverizer, ball mill or crusher;

[0061] S4, "single" supercritical: the improved gas disperser is used to remove the odor of sea cucumber coarse powder by supercritical extraction instrument. The sea cucumber coarse powder of step S3 is placed in the extraction kettle of the supercritical extraction instrument. The extraction kettle temperature is set to 30℃, the extraction pressure is 30MPa, the CO2 flow is 30L / h, and the extraction time is 2.5h. After extraction, the sea cucumber powder in the extraction kettle is collected and stored for later use;

[0062] S5, in-situ ultra-micro physical crushing: the sea cucumber powder obtained in step S4 is pre-cooled in liquid nitrogen for 1h, and then crushed by using an ultra-micro crusher. The rotation speed of the ultra-micro crusher is 12000rpm, the 0.6mm aperture screen is used for crushing, the powder bulk density is controlled to be 0.65g / mL, and the tap density is 0.73g / mL;

[0063] S6, "double" supercritical: the improved gas disperser is used to remove the odor of sea cucumber coarse powder by supercritical extraction instrument. The sea cucumber powder of step S5 is placed in the extraction kettle of the supercritical extraction instrument. The extraction kettle temperature is set to 30℃, the extraction pressure is 30MPa, the CO2 flow is 30L / h, and the extraction time is 2.5h. After extraction, the sea cucumber powder in the extraction kettle is collected and stored for later use;

[0064] S7, the sea cucumber powder obtained in step S6 is packaged, sterilized and sealed, and cooled to room temperature to obtain an ultra-micro sea cucumber powder product.

[0065] Example 2

[0066] S1, sea cucumber pretreatment: the captured sea cucumber is opened at the abdomen, the contents in the sea cucumber body cavity are removed, and the sea cucumber body wall is washed with water;

[0067] S2, sea cucumber nutrient enrichment: the pretreated sea cucumber obtained in step S1 is placed in warm water for temporary cultivation. The sea cucumber is placed in warm water at 60℃ for 6h. The water is changed every 0.5h to remove the salt in the sea cucumber. Then the sea cucumber is taken out and dried by a vacuum freeze dryer for 24h to obtain sea cucumber dry products;

[0068] S3, sea cucumber powder particle size regulation: sea cucumber dry products are crushed into 20 mesh sea cucumber coarse powder in a pulverizer, ball mill or crusher;

[0069] S4, "single" supercritical: the improved gas disperser is used to remove the odor of sea cucumber coarse powder by supercritical extraction instrument. The sea cucumber coarse powder of step S3 is placed in the extraction kettle of the supercritical extraction instrument. The extraction kettle temperature is set to 30℃, the extraction pressure is 30MPa, the CO2 flow is 30L / h, and the extraction time is 2.5h. After extraction, the sea cucumber powder in the extraction kettle is collected and stored for later use;

[0070] S5, in-situ ultra-micro physical pulverization: the sea cucumber powder obtained in step S4 is pre-cooled in liquid nitrogen for 1 h, and then pulverized by using an ultra-micro pulverizer, the rotation speed of the ultra-micro pulverizer is 12000 rpm, the 0.6 mm aperture screen is used for pulverization, the powder bulk density is controlled to be 0.70 g / mL, and the tap density is 0.80 g / mL;

[0071] S6, "double" supercritical: the improved gas disperser is used for deodorization of the sea cucumber powder, the sea cucumber powder obtained in step S5 is placed in the extraction kettle of the supercritical extraction instrument, the extraction kettle temperature is set to be 30℃, the extraction pressure is 30 MPa, the CO2 flow is 30 L / h, and the extraction time is 2.5 h; after the extraction is completed, the sea cucumber powder in the extraction kettle is collected and stored for standby;

[0072] S7, the sea cucumber powder obtained in step S6 is packaged, sterilized and sealed, and cooled to room temperature to obtain a sea cucumber powder product.

[0073] Example 3

[0074] S1, sea cucumber pretreatment: the captured sea cucumber is opened at the abdomen, the contents in the sea cucumber body cavity are removed, and the sea cucumber body wall is washed with water;

[0075] S2, sea cucumber nutrient enrichment: the pretreated sea cucumber obtained in step S1 is placed in warm water for temporary cultivation, the sea cucumber is placed in warm water at 60℃ for 6 h, the water is changed every 0.5 h to remove the salt in the sea cucumber, and then the sea cucumber is fished out and dried by a vacuum freeze dryer for 24 h to obtain a sea cucumber dry product;

[0076] S3, sea cucumber powder particle size control: the sea cucumber dry product is pulverized into 20-mesh sea cucumber coarse powder by using a pulverizer, a ball mill or a crusher;

[0077] S4, "single" supercritical: the improved gas disperser is used for deodorization of the sea cucumber coarse powder, the sea cucumber coarse powder obtained in step S3 is placed in the extraction kettle of the supercritical extraction instrument, the extraction kettle temperature is set to be 30℃, the extraction pressure is 30 MPa, the CO2 flow is 30 L / h, and the extraction time is 2.5 h; after the extraction is completed, the sea cucumber powder in the extraction kettle is collected and stored for standby;

[0078] S5, in-situ ultra-micro physical pulverization: the sea cucumber powder obtained in step S4 is pre-cooled in liquid nitrogen for 1 h, and then pulverized by using an ultra-micro pulverizer, the rotation speed of the ultra-micro pulverizer is 12000 rpm, the 0.6 mm aperture screen is used for pulverization, the powder bulk density is controlled to be 0.75 g / mL, and the tap density is 0.87 g / mL;

[0079] S6, "double" supercritical: using a supercritical extraction instrument equipped with an improved gas distributor to remove the odor of sea cucumber powder, placing the sea cucumber powder of step S5 in the extraction kettle of the supercritical extraction instrument, setting the extraction kettle temperature to 30°C, the extraction pressure to 30 MPa, the CO2 flow rate to 30 L / h, and the extraction time to 2.5 h; after the extraction is completed, the sea cucumber powder in the extraction kettle is collected and stored for use;

[0080] S7, packaging, sterilizing and sealing the sea cucumber powder obtained in step S6, and cooling to room temperature to obtain a sea cucumber powder product.

[0081] Comparative Example 1

[0082] S1, sea cucumber pretreatment: opening the abdomen of the captured sea cucumber, removing the contents in the sea cucumber body cavity, and washing the sea cucumber body wall with water to obtain a sea cucumber dry product;

[0083] S2, sea cucumber nutrient enrichment: placing the pretreated sea cucumber obtained in step S1 in warm water for temporary cultivation, placing the sea cucumber in warm water at 60°C for 6 h, changing the water every 0.5 h to remove salt from the sea cucumber, and then removing the sea cucumber through a vacuum freeze-drying machine for 24 h to remove water;

[0084] S3, sea cucumber powder particle size control: grinding the sea cucumber dry product into 20-mesh sea cucumber coarse powder in a grinder.

[0085] Comparative Example 2

[0086] S1, sea cucumber pretreatment: opening the abdomen of the captured sea cucumber, removing the contents in the sea cucumber body cavity, and washing the sea cucumber body wall with water to obtain a sea cucumber dry product, and then removing the sea cucumber through a vacuum freeze-drying machine for 24 h to remove water;

[0087] S2, sea cucumber powder particle size control: grinding the sea cucumber dry product into 20-mesh sea cucumber coarse powder in a grinder, ball mill or crusher.

[0088] According to the experimental method of Zhang Tuantong (Zhang Tuantong. Optimization of Determination Method of Sea Cucumber Polysaccharide and Its In Vivo Metabolism Research [D]. Dalian University of Technology, 2023.), 10 μL of sample solution was added to an enzyme-labeled plate, 200 μL of DMB staining solution (42.8 mg / L DMB solution was accurately prepared, and formic acid was used to adjust the pH to 3.3 to obtain A solution, and A solution was mixed with 2 mol / L Tris solution at a ratio of 10:1 to obtain DMB staining solution), shaken, and placed at room temperature for 15 min in the dark, and then the absorbance was measured at 525 nm. The content of sulfated polysaccharide in the sample was calculated according to the standard curve (y = 1.625x + 0.4439, R 2 = 0.9989).

[0089] The sea cucumber powder of Example 1, Example 2 and Comparative Example 1 was detected according to the above method, and the sea cucumber polysaccharide release rate results were as follows: Figure 5As shown, it can be seen that the release rate of sea cucumber polysaccharide is significantly improved after ultrafine grinding. Sea cucumber polysaccharide is mainly connected with protein, so it is difficult to be released into the digestive juice and then utilized by the human body, but the physical grinding technology provided in the embodiment can provide more enzyme cutting sites by changing the specific surface area and increasing the contact area with digestive enzymes, thereby improving the release rate of sea cucumber polysaccharide. Compared with Comparative Example 1, the polysaccharide release rate of Example 1 is increased by 21%.

[0090] With the gradual increase of the bulk density and the tap density, the particle size of the ultrafine sea cucumber powder gradually decreases, and the particle size of the ultrafine sea cucumber powder of Example 3 has reached a very small degree, which also means that it is already relatively difficult to process to this extent, and the polysaccharide release rate cannot be further improved. The polysaccharide release rate of the ultrafine sea cucumber powder prepared in Example 3 measured by the above experiment is not much different from that of Example 2. In addition, the microstructure experiment shows that with the decrease of the particle size, the electrostatic force increases, and the ultrafine powder is more prone to aggregation, which will affect the subsequent product processing and the food quality of the product. Therefore, the control of the bulk density and the tap density is best around 0.70 g / mL and 0.80 g / mL, and it is not suitable to be further improved. At this time, the polysaccharide release rate of the product is higher, the processing difficulty is not great, and the food quality is better.

[0091] Animal experiment content

[0092] 1. Experimental animals:

[0093] 36 APOE mice, male, 18-22 g; 6 C57 mice, male, 18-22 g; animal source Liaoning Changsheng Biotechnology Co., Ltd., experimental animal production license NO. SCXK (Liaoning) 2020-0001.

[0094] 2. Experimental grouping and treatment:

[0095] Experimental grouping and treatment:

[0096] (1) Normal control group: 8 C57 mice, normal diet, no gavage;

[0097] The background of the APOE mouse strain is C57 mice, and APOE is obtained by knocking out the APOE gene in the background of C57 mice. Generally, C57 mice are used as a blank control.

[0098] (2) Model group: 8 APOE mice, high-fat diet, no gavage;

[0099] (3) Positive control group: 8 APOE mice, high-fat diet, gavage of atorvastatin, 1.8 mg / kg, 300 uL once a day for 8 weeks;

[0100] (4) Comparative Example 1 High-dose group: 8 APOE mice were fed a high-fat diet and were given a high dose of ordinary sea cucumber powder by gavage, 2 mg / g, once a day for 8 weeks.

[0101] (5) Comparative Example 1 Low-dose group: 8 APOE mice were fed a high-fat diet and were given a low dose of ordinary sea cucumber powder by gavage, 1 mg / g, once a day for 8 weeks.

[0102] (6) Example 1 High-dose group: 8 APOE mice were fed a high-fat diet and were given a high dose of ultrafine sea cucumber powder by gavage, 2 mg / g, once a day for 8 weeks;

[0103] (7) Example 1 Low-dose group: 8 APOE mice were fed a high-fat diet and were given a high dose of ultrafine sea cucumber powder by gavage, 1 mg / g, once a day for 8 weeks;

[0104] Samples were collected after 8 weeks of gavage: serum (for 4 lipid tests), heart, liver, and aorta.

[0105] Referring to Cao Xu's experimental method (Cao Xu. Study on extraction process of total triterpenic acids from Ganoderma lucidum spores and exploration of its anti-atherosclerotic effects and mechanisms [D]. Xihua University, 2021.), blood samples should be placed at room temperature for 2 hours and then centrifuged at 3000 rpm for 15 minutes at 4℃. The supernatant can be collected for immediate testing.

[0106] Following the experimental method of Tian Guangjing (Tian Guangjing. Study on the ameliorative effect and mechanism of linseed oil on atherosclerosis [D]. Chinese Academy of Agricultural Sciences, 2018.), aortic tissue was dissected from mice and fixed in 4% paraformaldehyde. The aorta and liver were stained with hematoxylin and eosin (H&E) to assess pathological lesions.

[0107] A high-fat diet can lead to atherosclerosis and cause specific changes in cholesterol. The intervention effects of Example 1 and Comparative Example 1 on high-fat-induced atherosclerosis in mice were evaluated by measuring serum lipid levels in each group of mice. Figure 6 As shown. From Figure 6 As can be seen, the ultrafine sea cucumber powder of Example 1 can significantly reduce blood lipid levels in mice. This also confirms that the physical pulverization technology provided by the present invention does not alter the bioactivity of sea cucumber polysaccharides.

[0108] Histological changes in the aorta are key alterations in atherosclerosis, primarily manifested as arterial wall thickening and lipid accumulation. Example 1 and Comparative Example 1: aortic sections ( Figure 7 and wall thickness Figure 8). It can be seen that the mice fed with high-fat diet developed severe aortic tissue lesions, significant atherosclerotic plaque formation, a large number of foam cells formation and accumulation, protruding into the lumen, aortic intima thickening, and obvious endothelial cell and medial space. The aortic thickness of the control group was significantly increased, and the aortic thickness was 418 pm. Compared with Comparative Example 1, Example 1 had a better effect on atherosclerotic disease intervention.

[0109] Comparative Example 3

[0110] Referring to the preparation method in Example 1, only steps S1, S2, S3 and S4 are performed to prepare the superfine sea cucumber powder product.

[0111] Comparative Example 4

[0112] Referring to the preparation method in Example 1, only steps S1, S2, S3, S4 and S5 are performed to prepare the superfine sea cucumber powder product.

[0113] Comparative Example 5

[0114] Referring to the preparation method in Example 1, the supercritical extraction instrument equipped with an improved gas distributor is replaced by a conventional supercritical extraction instrument (the gas distributor is not improved and lacks the design of a second hole disc), and pipeline blockage occurs during the extraction process in step S6, and the extraction cannot be completed normally. This is because the superfine sea cucumber powder is too small in volume, which is easily carried out of the material basket by the carbon dioxide fluid and enters the carbon dioxide pipeline, causing blockage of the carbon dioxide pipeline. The sea cucumber powder that has not been extracted still has a heavy fishy smell and contains many odor impurities, and the purity of the product is not high.

[0115] Referring to the experimental method of Xu Jialin (Xu Jialin, Peng Jian, Xu Yujuan, et al. Physicochemical and powder properties of mango superfine powder with different peel addition amounts [J]. Food and Fermentation Industries, 2024, 1-13.), take an EP tube with a volume of 2 mL, fill the EP tube with sea cucumber powder, record the mass M of the sea cucumber powder in the container, and the calculation of the bulk density is as shown in the formula: bulk density (g / mL) = M / 2. The larger the bulk density value, the smaller the gap between particles and the smaller the particle size.

[0116] Referring to the experimental method of Li Zhenjiang (Li Zhenjiang, Liu Ying, Zhi Li, et al. Effect of ball milling superfine grinding on particle properties and polysaccharide extraction of inonotus obliquus powder [J]. China Food Additives, 2021, 32(11): 1-8.), take an EP tube with a volume of 2 mL, fill the EP tube with sea cucumber powder, continuously vibrate the EP tube, and continuously add samples until the powder weight reaches a stable state, record the mass M1 of the sea cucumber powder in the container, and the calculation of the tap density is as shown in the formula: tap density (g / mL) = M1 / 2. The larger the tap density, the smaller the gap between particles, the smaller the total volume, and the smaller the particle size.

[0117] Reference to the experimental method of Qibao Kun (Qibao Kun, Liu Yuyun, Yao Yuxue, et al. Analysis of the effect of protease on the volatile flavor of soybean meal based on HS-GC-IMS and HS-SPME-GC-MS [J]. Transactions of the Chinese Society of Agricultural Machinery, 2024, 1-27.) 0.1g sea cucumber powder, add 1mL deionized water, in 20mL headspace sampling bottle. Incubate at 60℃ for 30min, then automatically sample, sampling needle temperature is 85℃, sampling volume is 500μL, and non-diversion mode is used.

[0118] The chromatographic conditions are: WAX capillary column, column temperature 60℃, operation time 30min, carrier gas purity N2≥99.999%, N2purity≥99.999%. The analysis conditions are: 2mL / min 2min, 10mL / min 8min, 100mL / min 10min, 150mL / min 10min stop.

[0119] Qualitative analysis of different volatile substances: drift gas (nitrogen) is set to 150mL / min. Each spectrum is scanned for an average of 12 times. All analyses are performed in triplicate. With N-ketone C4-C9 (Beijing National Pharmaceutical Group Chemical Reagents Co., Ltd.) as an external reference, the retention index (RI) of VC is calculated. Qualitative analysis is performed by comparing the RI and the drift time of the standard in the GC-IMS library.

[0120] The ultra-fine sea cucumber powder prepared in Example 1, the sea cucumber powder prepared in Comparative Example 3 and Comparative Example 4 were detected, and the detection process referred to the above method. Among them, the GC-IMS two-dimensional difference topographic map is as shown in Figures 5-7 The darker the color, the greater the concentration of sample characteristic flavor. As can be seen from the figure, the green deodorization technology provided by the present application effectively removes the fishy substances in the sample, and the overall flavor of the sample after deodorization is not much different. The results of Comparative Example 3 and Comparative Example 4 can be seen that the ultra-fine grinding process will increase the concentration of volatile substances in the sample, so it is necessary to further deodorize after ultra-fine grinding, and the use of conventional supercritical extraction instrument cannot complete the deodorization of ultra-fine sea cucumber powder.

[0121] The above provided examples are not intended to limit the scope encompassed by the present application, and the described steps are not intended to limit the execution order. Those skilled in the art make obvious improvements to the present application in combination with existing common knowledge, which also falls within the protection scope defined by the claims of the present application.

Claims

1. A method for preparing a high polysaccharide release rate sea cucumber powder, characterized in that, The sea cucumber powder is prepared by the following steps: S1, sea cucumber pretreatment: opening the abdomen of sea cucumber, removing the contents in the body cavity of sea cucumber, and washing the body wall of sea cucumber with water to obtain pretreated sea cucumber; S2, sea cucumber nutrient enrichment: placing the pretreated sea cucumber obtained in step S1 in warm water for temporary cultivation, and then drying to obtain sea cucumber dry product; S3, sea cucumber powder particle size control: crushing the sea cucumber dry product obtained in step S2 into sea cucumber coarse powder with a particle size of 5-30; S4, "single" supercritical: placing the sea cucumber coarse powder obtained in step S3 in the extraction kettle of a supercritical extraction instrument, and performing supercritical carbon dioxide extraction to remove odor; S5, in-situ ultra-micro physical crushing: the sea cucumber powder deodorized in step S4 is subjected to ultra-micro crushing to obtain a powder, the bulk density of the powder is controlled to be 0.65-0.70 g / mL, and the tap density is controlled to be 0.70-0.80 g / mL; S6, "double" supercritical: placing the sea cucumber powder obtained in step S5 in the extraction kettle of a supercritical extraction instrument, and performing supercritical carbon dioxide extraction to remove odor to obtain sea cucumber powder with high polysaccharide release rate; In steps S4 and S6, the parameters of the extraction kettle are set to 30-40 °C, the extraction pressure is 25-30 MPa, the CO2 flow rate is 25-30 L / h, and the extraction time is 2-2.5 h; In steps S4 and S6, the supercritical extraction instrument is a supercritical extraction instrument equipped with a gas disperser; the gas disperser is an improved gas disperser; the improved gas disperser is placed in the extraction tank; The improved gas disperser comprises a mounting pipe (1), a connecting flange (2), a first hole disc (3), a filter screen (4), a support column (5), a baffle (6), a telescopic rod (7), a spring (70), a pressing plate (8), and a second hole disc (9); The mounting pipe (1) is made of transparent material for easy observation of the internal situation, the connecting flanges (2) are connected to the outer sides of the upper and lower parts of the mounting pipe (1) for easy connection of external pipelines, the first hole discs (3) are connected to the inner sides of the middle and lower parts of the mounting pipe (1), the filter screens (4) are connected to the lower sides of the first hole discs (3), the four support columns (5) are connected between the first hole discs (3), the baffles (6) are connected to the upper sides of the support columns (5) and are in contact with the mounting pipe (1), the telescopic rods (7) are connected to the upper sides of the baffles (6), the springs (70) are connected between the fixed ends and the telescopic ends of the telescopic rods (7), the pressing plates (8) are rotatably connected to the telescopic ends of the telescopic rods (7), the second hole disc (9) is placed between the baffles (6), the pressing plates (8) are in contact with the second hole disc (9), the first hole disc (3) and the second hole disc (9) have the same shape, and twelve conical parts are arranged on the upper side of the second hole disc (9) for easy backflow.

2. The production method according to claim 1, characterized by, In step S2, the warm water temporary cultivation treatment is as follows: placing the pretreated sea cucumber in warm water at 50-60 °C for 4-8 h, and replacing the warm water every 0.5-1 h.

3. The preparation method according to claim 1, characterized in that, The drying in step S2 includes freeze drying, heating drying, or air drying.

4. The preparation method according to claim 1, characterized in that, The pre-cooling is carried out before the pulverization in step S5, and the pre-cooling is carried out by using dry ice, liquid nitrogen or by placing in a frozen environment.

5. The production method as claimed in claim 1, characterized in that, The ultrafine pulverization in step S5 is carried out at a rotational speed of 8000-12000 rpm by using a 0.3-0.6 mm mesh sieve.

6. The production method as claimed in claim 1, characterized in that, The bulk density in step S5 is 0.67-0.70 g / mL, and the tap density is 0.77-0.80 g / mL.

7. A high polysaccharide releasing rate sea cucumber powder, characterized in that, The high polysaccharide release rate sea cucumber powder The preparation method according to any one of claims 1-6.

8. The high polysaccharide release rate sea cucumber powder according to claim 7 in the field of food or medicine preparation.

9. Use according to claim 8, characterised in that, The application in the field of medicine preparation is to prepare a medicine for treating atherosclerotic diseases.

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