Sea cucumber powder green processing technology capable of realizing directional enrichment and improving polysaccharide release rate

Abstract

The invention discloses a green processing technology of sea cucumber powder capable of realizing directional enrichment and improving polysaccharide release rate, and belongs to the field of food processing. The sea cucumber powder is prepared by the following steps: S1, pretreating sea cucumbers; s2, enrichment of nutrient substances of the sea cucumbers; s3, regulating and controlling the particle size of the sea cucumber powder; s4, performing'single 'supercritical treatment; s5, carrying out in-situ ultramicro physical crushing; and S6, performing'dual 'supercritical treatment. Through the design of the gas disperser and the additional arrangement of the second hole disc, the supercritical extraction instrument can perform effective supercritical carbon dioxide extraction on the ultramicro sea cucumber powder, the deodorization of the ultramicro sea cucumber powder is realized, and the eating quality of the ultramicro sea cucumber powder is improved. Moreover, by detecting and regulating the stacking density and the tap density, optimal processing parameters for the ultramicro sea cucumber powder are found, so that the prepared ultramicro sea cucumber powder is high in polysaccharide release rate, good in eating quality and easy for subsequent processing.

Description

Technical Field

[0001] The invention relates to a green processing technology for sea cucumber powder with directional enrichment and improved polysaccharide release rate, and belongs to the field of food processing. Background Art

[0002] Sea cucumber (Stichopus japonicus) belongs to cartilaginous invertebrates, Holothuria, Mollusca, benthic marine organisms, and is a valuable marine biological resource. Sea cucumbers are famous for their high protein and low fat nutritional characteristics, and are also rich in trace elements and a variety of amino acids. Among them, sea cucumber polysaccharides are one of the important active ingredients in the body wall of sea cucumbers, and their content accounts for 4% to 10% of the total organic matter in dry sea cucumbers. Sea cucumber polysaccharides are an acidic mucopolysaccharide that has strong biological activity in anticoagulation, lowering blood lipids, regulating immunity and anti-aging. However, sea cucumber polysaccharides are combined with proteins through glycopeptide bonds and hydrogen bonds, so the absorption and utilization of polysaccharides require the degradation of proteins connected to them.

[0003] In the prior art, in order to achieve effective separation of the two, the existing technology has proposed the concept of ultrafine sea cucumber powder, such as CN 106509694A--Preparation of ultrafine sea cucumber powder that is easy to eat and digest, which discloses that the digestibility of ultrafine sea cucumber powder is improved, and the crude polysaccharide content retention rate is high, with 95.42%; and CN 105495411 A--A method for preparing ultrafine pure powder slices of sea cucumber, which discloses that the ultrafine sea cucumber powder of 1-30μm reaches the specification of cell crushing, so that the active substances of sea cucumber are completely released. However, the processing technology of ultrafine sea cucumber powder in the above two patents is relatively simple. After simple washing, sterilization and enzyme inactivation, ultrafine powder is directly crushed. There is a lack of deodorization step, a large number of fishy molecules still remain in the product, and there is no indicator to measure the particle size of the crushed sea cucumber powder. Therefore, the edible quality of the final product is worrying, the product purity is poor, and the product quality is not easy to control.

[0004] Other methods for separating proteins and polysaccharides include protease hydrolysis and alkaline solution extraction, such as the method disclosed in patent CN 110204626A - a device and method for separating and refining sea cucumber polysaccharides. However, the use of alkaline solution will destroy the structure of sea cucumber polysaccharides, causing the detachment of glycosides and changes in the polysaccharide structure, so the concentration of the alkaline solution and the reaction conditions need to be strictly controlled. Enzymatic treatment requires control of the concentration, temperature and time of the enzyme reaction. The control conditions of this method are cumbersome, and the enzymatic reaction product has a bad flavor, which affects the taste. Therefore, it is better to directly perform physical crushing processing than to use enzymatic hydrolysis.

[0005] It can be seen that the extraction and utilization of sea cucumber polysaccharides still need further exploration in order to produce a sea cucumber polysaccharide product with good edible quality and excellent product quality, and use a more excellent and effective method. Summary of the invention

[0006] Technical issues

[0007] Sea cucumber polysaccharides have high use value, and if you want to use sea cucumber polysaccharides, you need to separate sea cucumber polysaccharides from proteins. Common separation methods include physical crushing and enzymatic extraction. 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 relatively crude, and the edible quality of the obtained sea cucumber powder is worrying, the product purity is poor, and the quality is difficult to control, so it needs to be improved.

[0008] Technical content

[0009] The purpose of the present invention is to overcome the shortcomings of the existing sea cucumber polysaccharide and protein separation technology and provide a method for preparing sea cucumber powder with a high polysaccharide release rate. The present invention improves the sea cucumber polysaccharide release rate by adopting appropriate physical crushing technology, prepares a sea cucumber powder with a high polysaccharide release rate and anti-atherosclerosis, and verifies it by using simulated digestion and animal models.

[0010] In order to achieve the above object, the present invention provides a method for preparing sea cucumber powder with a high polysaccharide release rate, wherein the sea cucumber powder is prepared by the following steps:

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

[0012] S2, enriching sea cucumber nutrients: placing the pretreated sea cucumber obtained in step S1 in warm water for temporary storage, and then drying to obtain a dried sea cucumber product;

[0013] S3, sea cucumber powder particle size control: the dried sea cucumber obtained in step S2 is crushed into 5-30 mesh sea cucumber coarse powder;

[0014] S4, "single" supercritical: placing the sea cucumber powder obtained in step S3 in the extraction kettle of a supercritical extractor, and extracting it with supercritical carbon dioxide to remove the fishy smell;

[0015] S5, in-situ ultrafine physical pulverization: ultrafine pulverize the sea cucumber powder removed from the fishy smell in step S4 to obtain a powder, and control 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: The sea cucumber powder obtained in step S5 is placed in the extraction kettle of a supercritical extractor, and subjected to supercritical carbon dioxide extraction to remove the fishy smell, thereby obtaining sea cucumber powder with a high polysaccharide release rate.

[0017] Furthermore, the temperature of the warm water in step S2 is 50-60°C.

[0018] Furthermore, the temporary maintenance time in step S2 is 4 to 8 hours.

[0019] Furthermore, the warm water temporary culture treatment in step S2 is as follows: placing the pretreated sea cucumber in warm water at 50-60° C. for 4-8 hours, and replacing the warm water every 0.5 hour to 1 hour.

[0020] Furthermore, the drying in step S2 includes freeze drying, heat drying or air drying.

[0021] Furthermore, the supercritical extractor in step S4 and step S6 is a supercritical extractor equipped with a gas diffuser.

[0022] Furthermore, the gas diffuser in the supercritical extraction apparatus is an improved gas diffuser.

[0023] Furthermore, the improved gas diffuser comprises a mounting pipe (1), a connecting flange (2), a first perforated plate (3), a filter screen (4), a support column (5), a baffle (6), a telescopic rod (7), a spring (70), a pressure plate (8) and a second perforated plate (9);

[0024] The mounting tube (1) is made of a transparent material, which is convenient for observing the internal situation. The outer sides of the upper and lower parts of the mounting tube (1) are connected with connecting flanges (2) to facilitate external pipe connection. The inner side of the middle part and the inner side of the lower part of the mounting tube (1) are connected with a first hole plate (3). The lower side of the first hole plate (3) is connected with a filter screen (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). The baffles (6) are connected to the mounting tube (1). ), the baffle plates (6) are connected with telescopic rods (7), a spring (70) is connected between the fixed end and the telescopic end of the telescopic rod (7), a pressure plate (8) is rotatably connected to the telescopic end of the telescopic rod (7), a second hole plate (9) is placed between the baffle plates (6), the pressure plate (8) is in contact with the second hole plate (9), the first hole plate (3) and the second hole plate (9) have the same shape, and twelve conical parts are provided on the upper side of the second hole plate (9) to facilitate backflow.

[0025] Furthermore, the parameters of the extraction kettle in step S4 are set to 30-40° C., extraction pressure to 25-30 MPa, CO2 flow rate to 25-30 L / h, and extraction time to 2-2.5 h.

[0026] Furthermore, in step S5, precooling is performed before pulverizing using an ultrafine pulverizer; the precooling is performed using dry ice, liquid nitrogen, or by placing the pulverizer in a freezing environment; the temperature of the freezing environment is -80 to -20 degrees Celsius.

[0027] Furthermore, in step S5, the ultrafine powder is crushed at a rotation speed of 8000 to 12000 rpm using a 0.3 to 0.6 mm pore size mesh screen.

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

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

[0030] Furthermore, the parameters of the extraction kettle in step S6 are set to 30-40° C., extraction pressure to 25-30 MPa, CO2 flow rate to 25-30 L / h, and extraction time to 2-2.5 h.

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

[0032] The sea cucumber powder with high polysaccharide release rate provided by the present invention is used in the field of food, health care product or medicine preparation.

[0033] Furthermore, the application in the field of health care products is to prepare health care products that help maintain healthy levels of blood lipids (cholesterol / triglycerides).

[0034] Furthermore, the application in the field of drug preparation is the preparation of drugs for preventing and treating atherosclerotic diseases.

[0035] The beneficial effects of the present invention are:

[0036] 1. The physical crushing process for protein separation of sea cucumber polysaccharides in the existing technology is relatively simple and lacks a deodorization step. The present invention designs a gas diffuser and adds a second hole plate design, so that the supercritical extractor can effectively perform supercritical carbon dioxide extraction on ultrafine sea cucumber powder, thereby achieving deodorization of ultrafine sea cucumber powder and improving the edible quality of ultrafine sea cucumber powder; and the design of the second hole plate can also allow the carbon dioxide fluid to reflux to achieve secondary extraction, thereby improving the extraction efficiency, effectively removing the fishy molecules in the ultrafine sea cucumber powder, and improving the purity of sea cucumber polysaccharides in the product.

[0037] 2. Currently, there is a lack of parameter measurement and quality control in the processing technology of ultrafine sea cucumber powder. The present invention finds the optimal processing parameters for ultrafine sea cucumber powder by detecting and regulating the bulk density and tap density, so that the ultrafine sea cucumber powder has a high polysaccharide release rate, good edible quality, and is easy for subsequent processing. In addition, the ultrafine sea cucumber powder has excellent effects of resisting atherosclerosis and maintaining healthy blood lipid levels, and is a product with broad market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The three-dimensional structure diagram of the improved gas diffuser; wherein, 1, mounting pipe, 2, connecting flange, 5, supporting column, 7, telescopic rod, 8, pressing plate, 9, second hole plate;

[0039] Figure 2 It is a partially cutaway three-dimensional structural schematic diagram of an improved gas diffuser; wherein, 1, a mounting pipe, 2, a connecting flange, 3, a first hole plate, 4, a filter screen, 5, a support column, 6, a baffle, 7, a telescopic rod, 8, a pressure plate, 9, a second hole plate;

[0040] Figure 3 The enlarged three-dimensional structural diagram of the improved gas diffuser at A; wherein, 1, mounting tube, 5, support column, 6, baffle, 7, telescopic rod, 70, spring, 8, pressure plate, 9, second hole plate;

[0041] Figure 4 This is a supercritical extraction device equipped with a gas diffuser, where the red arrow indicates the installation position of the improved gas diffuser;

[0042] Figure 5 The release curves of sea cucumber polysaccharides in Examples 1 and 2 and Comparative Example 1;

[0043] Figure 6 The cholesterol levels in the blood of mice fed with high fat diet after intervention in Example 1 and Comparative Example 1 (different letters indicate significance);

[0044] Figure 7 The aorta slices of mice fed with high fat diet after intervention in Example 1 and Comparative Example 1;

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

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

[0047] Fig.10 is a comparative example 3GC-IMS two-dimensional difference topography;

[0048] Fig.11 This is a comparative example 4 GC-IMS two-dimensional difference topography map. DETAILED DESCRIPTION

[0049] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0050] In the present invention, unless otherwise clearly specified and limited, the terms "connected", "connected", and "fixed" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0051] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, a first feature being "above", "above" and "above" a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature. A first feature being "below", "below" and "below" a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.

[0052] Experimental instruments

[0053] The supercritical extractor equipped with an improved gas diffuser used in the embodiments and comparative examples is manufactured by Jiangsu Gaoke Pharmaceutical Equipment Co., Ltd. and has a model number of GKSCFE220-40-30L. The gas diffuser of the supercritical extractor has been improved. Figures 1 to 3 .

[0054] The improved gas diffuser comprises a mounting pipe 1, a connecting flange 2, a first perforated plate 3, a filter screen 4, a supporting column 5, a baffle 6, a telescopic rod 7, a spring 70, a pressing plate 8 and a second perforated plate 9.

[0055] The mounting tube 1 is made of transparent material, which is convenient for observing the internal situation. The outer sides of the upper and lower parts of the mounting tube 1 are connected with connecting flanges 2 for external connection to pipelines. The inner side of the middle part and the inner side of the lower part of the mounting tube 1 are connected with a first hole plate 3, and the lower side of the first hole plate 3 is connected with a filter screen 4. Four support columns 5 are connected between the first hole plates 3, and the upper sides of the support columns 5 are connected with baffles 6, and the baffles 6 are in contact with the mounting tube 1. The baffles 6 are connected with telescopic rods 7, and springs 70 are connected between the fixed end and the telescopic end of the telescopic rod 7. The telescopic ends of the telescopic rods 7 are rotatably connected with pressure plates 8. A second hole plate 9 is placed between the baffles 6, and the pressure plates 8 are in contact with the second hole plate 9. The first hole plate 3 and the second hole plate 9 have the same shape, and twelve conical parts are provided on the upper side of the second hole plate 9 for reflux.

[0056] When supercritical extraction of carbon dioxide is performed, an improved gas diffuser is used to drive the device to the place where the components need to be extracted, so that the connecting flange 2 at the bottom of the mounting tube 1 is connected to the feed pipe. The mounting tube 1 is made of transparent material, which is convenient for observing the internal situation. Then, the pressing plate 8 is pulled to move and rotate, so that the telescopic rod 7 is extended, and the spring 70 is stretched. Then, the second orifice plate 9 is brought into contact with the baffle 6, and then the pressing plate 8 is reversely rotated to reset, and then the pressing plate 8 is released, the spring 70 rebounds, and the telescopic rod 7 shrinks and recovers, so that the pressing plate 8 is brought into contact with the second orifice plate 9, so that the second orifice plate 9 can be fixed to prevent the second orifice plate 9 from falling when the carbon dioxide fluid flows. Then, it is connected to the feed port of the extraction tank through the upper connecting flange 2. By adding carbon dioxide fluid into the feed pipeline, 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 have the same shape, so that the carbon dioxide fluid can be fed evenly, the extraction efficiency and the production efficiency can be improved. At the same time, the device can be placed in the extraction tank, and the carbon dioxide can achieve a reflux effect under the action of the cone, so that the carbon dioxide fluid refluxes under the action of the cone on the second hole plate 9 for secondary extraction, which is convenient for secondary extraction of the material, reduces the pressure of the carbon dioxide fluid when it is discharged from the material basket, prevents the material from being brought out, and improves the extraction effect.

[0057] Example 1

[0058] S1. Pretreatment of sea cucumber: opening the abdomen of fresh sea cucumber, removing the contents in the body cavity of sea cucumber, and washing the body wall of sea cucumber with water;

[0059] S2, enrichment of sea cucumber nutrients: placing the pretreated sea cucumber obtained in step S1 in warm water at 60°C for 6 hours, changing the water every 0.5 hours to remove the salt in the sea cucumber, and then removing the sea cucumber and removing the water in a vacuum freeze dryer for 24 hours to obtain a dried sea cucumber product;

[0060] S3, sea cucumber powder particle size control: the dried sea cucumber is crushed into 20 mesh sea cucumber coarse powder in a grinder, ball mill or crusher;

[0061] S4, "single" supercritical: use a supercritical extractor equipped with an improved gas diffuser to remove the fishy smell of the sea cucumber powder, place the sea cucumber powder in step S3 in an extraction kettle of the supercritical extractor, the extraction kettle temperature is set to 30°C, the extraction pressure is 30MPa, the CO2 flow rate is 30L / h, and the extraction time is 2.5h; after the extraction is completed, collect the sea cucumber powder in the extraction kettle and save it for later use;

[0062] S5, in-situ ultrafine physical pulverization: the sea cucumber powder obtained in step S4 is precooled in liquid nitrogen for 1 hour, and then pulverized by an ultrafine pulverizer, the ultrafine pulverizer speed is 12000rpm, and the 0.6mm aperture mesh screen is used for pulverization, and the bulk density of the powder is controlled to be 0.65g / mL, and the tap density is 0.73g / mL;

[0063] S6, "double" supercritical: Use a supercritical extractor equipped with an improved gas diffuser to remove the fishy smell of the sea cucumber powder, place the sea cucumber powder in step S5 in an extraction kettle of the supercritical extractor, set the extraction kettle temperature to 30°C, the extraction pressure to 30MPa, the CO2 flow rate to 30L / h, and the extraction time to 2.5h; after the extraction is completed, collect the sea cucumber powder in the extraction kettle and store it for later use;

[0064] S7, packaging, sterilizing, and sealing the sea cucumber powder obtained in step S6, and cooling to room temperature to obtain an ultrafine sea cucumber powder product.

[0065] Example 2

[0066] S1. Sea cucumber pretreatment: opening the abdomen of the captured sea cucumber, removing the contents in the body cavity of the sea cucumber, and washing the body wall of the sea cucumber with water;

[0067] S2, enrichment of sea cucumber nutrients: the pretreated sea cucumber obtained in step S1 is placed in warm water for temporary storage, the sea cucumber is placed in 60°C warm water for 6 hours, the water is changed every 0.5 hours to remove the salt in the sea cucumber, and then the sea cucumber is taken out and the water is removed by a vacuum freeze dryer for 24 hours to obtain a dried sea cucumber product;

[0068] S3, sea cucumber powder particle size control: the dried sea cucumber is crushed into 20 mesh sea cucumber coarse powder in a grinder, ball mill or crusher;

[0069] S4, "single" supercritical: use a supercritical extractor equipped with an improved gas diffuser to remove the fishy smell of the sea cucumber powder, place the sea cucumber powder in step S3 in an extraction kettle of the supercritical extractor, the extraction kettle temperature is set to 30°C, the extraction pressure is 30MPa, the CO2 flow rate is 30L / h, and the extraction time is 2.5h; after the extraction is completed, collect the sea cucumber powder in the extraction kettle and save it for later use;

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

[0071] S6, "double" supercritical: Use a supercritical extractor equipped with an improved gas diffuser to remove the fishy smell of the sea cucumber powder, place the sea cucumber powder in step S5 in an extraction kettle of the supercritical extractor, set the extraction kettle temperature to 30°C, the extraction pressure to 30MPa, the CO2 flow rate to 30L / h, and the extraction time to 2.5h; after the extraction is completed, collect the sea cucumber powder in the extraction kettle and store it for later use;

[0072] S7. Pack, sterilize, and seal the sea cucumber powder obtained in step S6, and cool to room temperature to obtain a sea cucumber powder product.

[0073] Example 3

[0074] S1. Sea cucumber pretreatment: opening the abdomen of the captured sea cucumber, removing the contents in the body cavity of the sea cucumber, and washing the body wall of the sea cucumber with water;

[0075] S2, enrichment of sea cucumber nutrients: the pretreated sea cucumber obtained in step S1 is placed in warm water for temporary storage, the sea cucumber is placed in 60°C warm water for 6 hours, the water is changed every 0.5 hours to remove the salt in the sea cucumber, and then the sea cucumber is taken out and the water is removed by a vacuum freeze dryer for 24 hours to obtain a dried sea cucumber product;

[0076] S3, sea cucumber powder particle size control: the dried sea cucumber is crushed into 20 mesh sea cucumber coarse powder in a grinder, ball mill or crusher;

[0077] S4, "single" supercritical: use a supercritical extractor equipped with an improved gas diffuser to remove the fishy smell of the sea cucumber powder, place the sea cucumber powder in step S3 in an extraction kettle of the supercritical extractor, the extraction kettle temperature is set to 30°C, the extraction pressure is 30MPa, the CO2 flow rate is 30L / h, and the extraction time is 2.5h; after the extraction is completed, collect the sea cucumber powder in the extraction kettle and save it for later use;

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

[0079] S6, "double" supercritical: Use a supercritical extractor equipped with an improved gas diffuser to remove the fishy smell of the sea cucumber powder, place the sea cucumber powder in step S5 in an extraction kettle of the supercritical extractor, set the extraction kettle temperature to 30°C, the extraction pressure to 30MPa, the CO2 flow rate to 30L / h, and the extraction time to 2.5h; after the extraction is completed, collect the sea cucumber powder in the extraction kettle and store it for later use;

[0080] S7. Pack, sterilize, and seal the sea cucumber powder obtained in step S6, and cool to room temperature to obtain a sea cucumber powder product.

[0081] Comparative Example 1

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

[0083] S2, enrichment of sea cucumber nutrients: the pretreated sea cucumber obtained in step S1 is placed in warm water for temporary storage, the sea cucumber is placed in 60° C. warm water for 6 hours, the water is changed every 0.5 hours to remove the salt in the sea cucumber, and then the sea cucumber is taken out and passed through a vacuum freeze dryer for 24 hours to remove moisture;

[0084] S3. Sea cucumber powder particle size control: The dried sea cucumber is crushed into 20-mesh sea cucumber coarse powder in a grinder.

[0085] Comparative Example 2

[0086] S1, sea cucumber pretreatment: open the abdomen of the fresh sea cucumber, remove the contents in the sea cucumber body cavity, and wash the sea cucumber body wall with water to obtain a dried sea cucumber, then remove the sea cucumber and pass it through a vacuum freeze dryer for 24 hours to remove moisture;

[0087] S2. Sea cucumber powder particle size control: The dried sea cucumber is crushed into 20-mesh sea cucumber coarse powder in a grinder, ball mill or crusher.

[0088] Referring to Zhang Tongtong's experimental method (Zhang Tongtong. Optimization of the determination method of sea cucumber polysaccharides and its in vivo metabolism study [D]. Dalian University of Technology, 2023.), 10 μL of the sample solution was added to the ELISA plate, and then 200 μL of DMB staining solution was added (accurately prepare 42.8 mg / L DMB solution, and adjust the pH to 3.3 with formic acid to obtain solution A, and mix solution A with 2 mol / L Tris solution at a ratio of 10:1 to obtain DMB staining solution), shake, place at room temperature away from light for 15 minutes, and then measure the absorbance at 525 nm. The sulfated polysaccharide content in the sample was calculated using 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 tested as above, and the results of the sea cucumber polysaccharide release rate were as follows: Figure 5As shown, it can be seen that the release rate of sea cucumber polysaccharides is significantly improved after ultrafine grinding. Sea cucumber polysaccharides are mainly connected to proteins, so it is difficult to be released into the digestive juice and then used by the human body. However, the physical grinding technology provided in the embodiment can increase the contact area with digestive enzymes by changing its specific surface area, providing more enzyme cleavage sites, and improving the release rate of sea cucumber polysaccharides. Compared with Comparative Example 1, the polysaccharide release rate of Example 1 is increased by 21%.

[0090] As the bulk density and tap density gradually increase, the particle size of the ultrafine sea cucumber powder gradually decreases. The particle size of the ultrafine sea cucumber powder in Example 3 has reached a very small level, which also means that it is difficult to process to this level, and the polysaccharide release rate cannot be further improved. The polysaccharide release rate of the ultrafine sea cucumber powder obtained in Example 3 measured by the above experiment is not much different from the result of Example 2. In addition, the microstructure experiment shows that as the particles are reduced, the electrostatic force increases, and the ultrafine powder is easier to aggregate, which will affect the subsequent product processing and the edible quality of the product. Therefore, the control of the bulk density and tap density to about 0.70g / mL and 0.80g / mL is the best effect, and it is not suitable to be further increased. At this time, the polysaccharide release rate of the product is high, the processing difficulty is not great, and the edible 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.: NO.SCXK(Liao)2020-0001.

[0094] 2. Experimental grouping and treatment:

[0095] Experimental groups and treatments:

[0096] (1) Normal control group: 8 C57 mice, fed a normal diet without oral gavage;

[0097] The strain background of APOE mice is C57 mice. APOE is obtained after knocking out the APOE gene in the background of C57 mice. Generally, C57 mice are used as blank controls.

[0098] (2) Model group: 8 APOE mice, fed with a high-fat diet but not gavage;

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

[0100] (4) Comparative Example 1 High-dose group: 8 APOE mice, fed with a high-fat diet, were gavaged with a high-dose of common sea cucumber powder, 2 mg / g, once a day for 8 weeks;

[0101] (5) Comparative Example 1 low-dose group: 8 APOE mice, fed with a high-fat diet, were gavaged with a low dose of common sea cucumber powder, 1 mg / g, once a day for 8 weeks;

[0102] (6) Example 1 high-dose group: 8 APOE mice, fed with a high-fat diet, gavaged with high-dose ultrafine sea cucumber powder, 2 mg / g, once a day for 8 weeks;

[0103] (7) Example 1 low-dose group: 8 APOE mice, fed with a high-fat diet, gavaged with high-dose ultrafine sea cucumber powder, 1 mg / g, once a day for 8 weeks;

[0104] After 8 weeks of intragastric administration, samples were collected: serum (4 blood lipid tests), heart, liver, and aorta.

[0105] Referring to Cao Xu's experimental method (Cao Xu. Research on the extraction process of total triterpene acids from Ganoderma lucidum spores and exploration of its anti-atherosclerotic effects and mechanisms [D]. Xihua University, 2021.), please place the blood sample at room temperature for 2 hours, then centrifuge it at 4℃ and 3000 rpm for 15 minutes, and take the supernatant for immediate testing.

[0106] Referring to the experimental method of Tian Guangjing (Tian Guangjing. Study on the improvement effect of flaxseed oil on atherosclerosis and its mechanism [D]. Chinese Academy of Agricultural Sciences, 2018.), aortic tissues were dissected from mice and fixed in 4% paraformaldehyde. The aorta and liver parts were stained with hematoxylin and eosin (H&E) to evaluate 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, and the serum lipid levels of mice in each group were measured, such as Figure 6 As shown. Figure 6 It can be seen that the ultrafine sea cucumber powder of Example 1 can significantly reduce the blood lipid level of mice. This also confirms that the physical crushing technology provided by the present invention will not change the biological activity of sea cucumber polysaccharides.

[0108] The histological changes of aorta are the key changes of atherosclerosis, which are mainly manifested as thickening of arterial wall and accumulation of vascular lipids. Figure 7 ) and wall thickness ( Figure 8). It can be seen that mice fed with high-fat diet had severe aortic tissue lesions, significant atherosclerotic plaque formation, a large number of foam cells formed and accumulated, protruding into the lumen, thickening of the aortic intima, and obvious gaps between endothelial cells and media. The aortic thickness of the control group increased significantly, with a thickness of 418 μm. Compared with comparative example 1, example 1 has a better intervention effect on atherosclerotic disease.

[0109] Comparative Example 3

[0110] Referring to the preparation method in Example 1, only steps S1, S2, S3 and S4 are performed to obtain the ultrafine 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 obtain an ultra-fine sea cucumber powder product.

[0113] Comparative Example 5

[0114] With reference to the preparation method in Example 1, the supercritical extractor equipped with an improved gas diffuser is replaced by a conventional supercritical extractor (the gas diffuser is not improved, and the design of the second orifice disk is lacking), and pipeline blockage occurs during the extraction process of step S6, and extraction cannot be normally completed. This is because the volume of the supermicro sea cucumber powder is too small, and it is easy to be taken out of the material basket by the carbon dioxide fluid, enter the carbon dioxide pipeline, and cause the blockage of the carbon dioxide pipeline. The sea cucumber powder that has not been extracted still has a heavier fishy smell, and many odor impurities remain, and the purity of the product is not high.

[0115] Referring to Xu Jialin's experimental method (Xu Jialin, Peng Jian, Xu Yujuan, et al. Study on the physical and chemical properties of mango ultrafine powder with different amounts of peel added [J]. Food and Fermentation Industry, 2024, 1-13.), take an EP tube with a volume of 2mL, fill the EP tube with sea cucumber powder, and record the mass M of the sea cucumber powder in the container. The calculation of the bulk density is 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 ultrafine grinding on particle characteristics of birch inonotus powder and polysaccharide extraction effect [J]. Chinese Food Additives, 2021, 32(11): 1-8.), take a 2mL EP tube, fill the EP tube with sea cucumber powder, vibrate the EP tube continuously, and continue to add samples until the powder weight reaches a stable state. Record the mass M1 of the sea cucumber powder in the container. The calculation of the tap density is shown in the formula: tap density (g / mL) = M1 / 2. The larger the tap density, the smaller the particle gap, the smaller the total volume, and the smaller the particle size.

[0117] Referring to the experimental method of Qi Baokun (Qi Baokun, Liu Yuwen, Yao Yuxue, et al. Analysis of the influence 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 of sea cucumber powder was added with 1mL of deionized water and placed in a 20mL headspace injection bottle. After incubation at 60℃ for 30min, the sample was automatically injected, the injection needle temperature was 85℃, the injection volume was 500μL, and the splitless mode was adopted.

[0118] Chromatographic conditions: WAX capillary column, column temperature 60°C, operation time 30 min, carrier gas purity N2 ≥ 99.999%, N2 purity ≥ 99.999%. Analysis conditions: 2 mL / min 2 min, 10 mL / min 8 min, 100 mL / min 10 min, 150 mL / min 10 min stop.

[0119] Qualitative analysis of different volatile substances: The drift gas (nitrogen) was set to 150 mL / min. Each spectrum was scanned 12 times on average. All analyses were performed in triplicate. The retention index (RI) of VC was calculated using N-ketone C4-C9 (Beijing Sinopharm Chemical Reagent Co., Ltd.) as an external reference. Qualitative analysis was performed by comparing the RI with the drift time of the standard in the GC-IMS library.

[0120] The ultrafine sea cucumber powder prepared in Example 1 and the sea cucumber powder prepared in Comparative Examples 3 and 4 were tested, and the testing process was referred to the above method. Among them, the GC-IMS two-dimensional difference topography is as follows Figures 5 to 7 As shown, the darker the color, the greater the concentration of the characteristic flavor of the sample. It can be seen from the figure that the green deodorization technology provided by the present invention 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 Examples 3 and 4 show that the ultrafine grinding process will increase the concentration of volatile substances in the sample, so it is necessary to further deodorize after ultrafine grinding, and the use of conventional supercritical extraction instruments cannot complete the deodorization of ultrafine sea cucumber powder.

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

Claims

1. A method for preparing sea cucumber powder with high polysaccharide release rate, characterized in that: The sea cucumber powder is prepared by the following steps: S1, pretreatment of sea cucumber: opening the abdomen of the sea cucumber, removing the contents in the body cavity of the sea cucumber, and washing the body wall of the sea cucumber with water to obtain the pretreated sea cucumber; S2, enriching sea cucumber nutrients: placing the pretreated sea cucumber obtained in step S1 in warm water for temporary storage, and then drying to obtain a dried sea cucumber product; S3, sea cucumber powder particle size control: the dried sea cucumber obtained in step S2 is crushed into 5-30 mesh sea cucumber coarse powder; S4, "single weight" supercritical: the sea cucumber powder obtained in step S3 is placed in the extraction kettle of a supercritical extractor, and subjected to supercritical carbon dioxide extraction to remove the fishy smell; S5, in-situ ultrafine physical pulverization: ultrafine pulverize the sea cucumber powder removed from the fishy smell in step S4 to obtain a powder, and control 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; S6, "double" supercritical: placing the sea cucumber powder obtained in step S5 in an extraction kettle of a supercritical extractor, performing supercritical carbon dioxide extraction to remove the fishy smell, and obtaining sea cucumber powder with a high polysaccharide release rate; The supercritical extractor in step S4 and step S6 is a supercritical extractor equipped with a gas diffuser; the gas diffuser is an improved gas diffuser; The improved gas diffuser comprises a mounting pipe (1), a connecting flange (2), a first perforated plate (3), a filter screen (4), a supporting column (5), a baffle (6), a telescopic rod (7), a spring (70), a pressure plate (8) and a second perforated plate (9); The mounting tube (1) is made of a transparent material, which is convenient for observing the internal situation. The outer sides of the upper and lower parts of the mounting tube (1) are connected with connecting flanges (2) to facilitate external pipe connection. The inner side of the middle part and the inner side of the lower part of the mounting tube (1) are connected with a first hole plate (3). The lower side of the first hole plate (3) is connected with a filter screen (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). The baffles (6) are connected to the mounting tube (1). ), the baffle plates (6) are connected with telescopic rods (7), a spring (70) is connected between the fixed end and the telescopic end of the telescopic rod (7), a pressure plate (8) is rotatably connected to the telescopic end of the telescopic rod (7), a second hole plate (9) is placed between the baffle plates (6), the pressure plate (8) is in contact with the second hole plate (9), the first hole plate (3) and the second hole plate (9) have the same shape, and twelve conical parts are provided on the upper side of the second hole plate (9) to facilitate backflow.

2. The preparation method according to claim 1, characterized in that: The warm water temporary culture treatment in step S2 is as follows: placing the pretreated sea cucumber in warm water at 50-60° C. for 4-8 hours, and replacing the warm water every 0.5 hour to 1 hour.

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

4. The preparation method according to claim 1, characterized in that: The parameters of the extraction kettle in step S4 and step S6 are set to 30-40° C., extraction pressure to 25-30 MPa, CO2 flow rate to 25-30 L / h, and extraction time to 2-2.5 h.

5. The preparation method according to claim 1, characterized in that: In step S5, precooling is performed before pulverizing using an ultrafine pulverizer; the precooling is performed using dry ice, liquid nitrogen, or by placing the pulverizer in a freezing environment.

6. The preparation method according to claim 1, characterized in that: In step S5, the ultrafine powder is crushed at a rotation speed of 8000 to 12000 rpm using a 0.3 to 0.6 mm pore size mesh screen.

7. The preparation method according to claim 1, characterized in that: In step S5, the bulk density is 0.67-0.70 g / mL, and the tap density is 0.77-0.80 g / mL.

8. A sea cucumber powder with high polysaccharide release rate, characterized in that: The sea cucumber powder with high polysaccharide release rate Prepared according to the preparation method according to any one of claims 1 to 7.

9. Use of the sea cucumber powder with high polysaccharide release rate as claimed in claim 8 in the field of food, health care products or medicine preparation.

10. The use according to claim 9, characterized in that The application in the field of health care products is to prepare health care products that help maintain healthy blood lipid levels; the application in the field of drug preparation is to prepare drugs for treating atherosclerotic diseases.

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