Preparation method and device of SOD (superoxide dismutase) capable of resisting pepsin degradation

Abstract

The invention belongs to the field of SOD preparation, and discloses a pepsin degradation resistant SOD preparation method and device, the method comprises the following steps: S1, powder preparation: dissolving SOD fermentation pure liquid in a buffer solution with the pH value of 5.2-7.8, sequentially adding a protective agent and theaflavin, stirring, filtering, and putting into a freeze dryer to prepare freeze-dried powder; s2, drying; s3, tabletting; s4, preparing a coating solution, namely dissolving wheat gliadin and soybean protein in a mixed solution prepared from water and ethanol in a volume ratio of 1: 1, adding a plasticizer, stirring for 20 minutes by using a magnetic stirrer, and performing ultrasonic treatment for 40 seconds, wherein the mass ratio of the wheat gliadin to the soybean protein is (0.5-1): 1; and S5, coating: placing the tablets in a coating machine, setting the air inlet rate, the air inlet temperature, the pot rotating speed, the atomization rate, the sector pressure and the guniting flow until the weight of the coating is increased to 3-10% by mass and the thickness of the coating is 20-50 microns, and drying and cooling. The method has the beneficial effects that the prepared SOD is not easy to be photolyzed, and can resist gastric acid degradation, so that the complete activity of the SOD is ensured.

Description

Technical Field

[0001] The present invention belongs to the technical field of SOD preparation, and particularly relates to a method and device for preparing SOD that resists degradation by pepsin. Background Art

[0002] As an important free radical scavenger in organisms, SOD (superoxide dismutase) has important application values in aspects such as antioxidant and preventing organism aging. Since its discovery, SOD has been favored by scholars at home and abroad, and the research on the properties of SOD has become increasingly in-depth. It is found that SOD can scavenge free radicals, improve the redox environment in the intestine, inhibit the growth and reproduction of harmful bacteria, reduce their adhesion and invasion to the intestinal mucosa, thereby maintaining the stability of the intestinal flora; at the same time, it can also create a suitable growth environment for beneficial bacteria such as Bifidobacterium and Lactobacillus by regulating the redox potential in the intestine, promote their growth and reproduction, and the beneficial bacteria can produce substances such as short-chain fatty acids, provide energy for intestinal epithelial cells, enhance the barrier function of the intestinal mucosa, and further maintain intestinal health.

[0003] SOD is essentially a kind of protein, with poor light and heat stability and easy inactivation under acid-base conditions. Therefore, in the gastric acid environment, its spatial structure may denature, resulting in a decrease or even loss of activity, and pepsin in the stomach will decompose the protein into polypeptides and amino acids, making it difficult for SOD to maintain a complete active structure to reach the intestine to play a role; at the same time, it is easily affected by light and temperature during tablet preparation, resulting in photodegradation or inactivation, and poor stability.

[0004] When using the existing rotary tablet press to feed powder during the preparation of SOD tablets, due to continuous feeding and intervals between the die slots, the powder will be fed outside the die slots and accumulate in a triangular shape in the die slots during feeding. When pressing tablets, too much powder will be squeezed onto the workbench, resulting in serious powder waste and low tablet preparation efficiency. At the same time, the powder accumulation will also cause uneven density and inconsistent strength in various parts of the prepared tablets. Summary of the Invention

[0005] The purpose of the present invention is to provide a method and device for preparing SOD that resists degradation by pepsin, which has the effects of being not easily photolyzed, being able to resist gastric acid degradation, and ensuring the complete activity of SOD.

[0006] The above technical purpose of the present invention is achieved through the following technical solutions:

[0007] A method for preparing SOD that resists degradation by pepsin includes the following steps:

[0008] S1. Powder preparation: Dissolve the SOD fermentation pure liquid with a purity greater than 95% in a buffer solution with a pH of 5.2 - 7.8. Add a protective agent and theaflavin in sequence, stir for 30 min, filter, and then put it into a freeze dryer. First, reduce the temperature at a rate of 1 - 3 °C / min to 40 °C to -60 °C and maintain for 2 - 4 h. Then start the freeze dryer and freeze for 12 - 48 h under the conditions of -50 °C to -30 °C and a high vacuum of 10 - 50 Pa to obtain the freeze-dried powder.

[0009] S2. Drying: After S1 is completed, gradually increase the temperature to 0 °C - 25 °C, maintain the vacuum degree at 10 - 30 Pa for 2 - 10 h until the water content of the freeze-dried powder is reduced to less than 5%.

[0010] S3. Tabletting: Mix the SOD freeze-dried powder and the binder evenly. The mass ratio of the binder to the SOD freeze-dried powder is 13 - 20:20. Then use a rotary tabletting machine to tablet it into tablets with a diameter of 3 - 8 mm;

[0011] S4. Coating solution preparation: Dissolve gliadin and soy protein in a mixed solution prepared with a volume ratio of water:ethanol of 1:1. Among them, the mass ratio of gliadin to soy protein is 0.5 - 1:1. Add a plasticizer and stir with a magnetic stirrer at room temperature and a speed of 500 rpm for 20 min, and then ultrasonicate for 40 s with an ultrasonic device;

[0012] S5. Coating: Place the tablets in a coating machine, set the air inlet volume at 1500 - 1800 m 3 / h, the air inlet temperature at 35 - 40 °C, the pan rotation speed at 2 - 10 rpm, set the atomization rate at 2 - 5 mL / min, the fan surface pressure at 1.0 - 3.0 bar, and the spraying flow rate at 60 - 400 g / min until the coating weight gain reaches a mass fraction of 3% - 10% and the coating thickness is 20 - 50 μm. Slowly dry at 40 °C for 1 h and cool to obtain the coated tablets.

[0013] A further setting of the present invention is that in the step S1, the specific protective agent is one or more of trehalose, cyclodextrin, and mannitol. The volume ratio of the protective agent to the fermentation pure liquid is 1:1, and the volume ratio of theaflavin to the fermentation pure liquid is 1 - 3:1.

[0014] A further setting of the present invention is that in the step S2, the binder is one or more of sorbitol and maltodextrin.

[0015] A further setting of the present invention is that in the step S3, gliadin can be replaced by zein.

[0016] A further setting of the present invention is that in the step S3, the plasticizer is glycerol or polyethylene glycol 400, accounting for 10% - 15% of the mass of gliadin.

[0017] A preparation device for pepsin-resistant SOD, applying the above-mentioned preparation method for pepsin-resistant SOD, includes a rotary tablet press. The rotary tablet press includes a workbench, a material guiding groove is opened on the workbench, a chassis is fixedly arranged in the material guiding groove, a turntable is rotatably abutted above the chassis, mold grooves are evenly arranged along the circumference of the turntable, there are at least four mold grooves and a waste material groove is arranged between each mold groove. A material storage space is formed between the mold groove and the chassis. Along the outer circumference of the turntable, a feeding component, a leveling component, a pressing component and a demolding component are arranged in sequence. A blanking through groove is opened at the position of the chassis where the demolding component is located. The waste material groove is inclined downward and a conical cylinder is arranged below it. The top of the conical cylinder is open. A rotating shaft is fixedly arranged at the center of the turntable. The rotating shaft penetrates through the conical cylinder and an auxiliary blanking component is arranged at the part of the rotating shaft located inside the conical cylinder;

[0018] The auxiliary blanking component includes a supporting cross bar fixedly arranged on the rotating shaft. A scraping plate is fixedly arranged at one end of the supporting cross bar away from the rotating shaft. The bottom of the scraping plate abuts against the inner wall of the conical cylinder for scraping materials.

[0019] The leveling component includes an L-shaped fixing frame fixed on the workbench. A leveling plate is arranged on the L-shaped fixing frame. The leveling plate is arc-shaped and the opening faces the feeding component. The bottom of the leveling plate is slidably connected with the turntable;

[0020] It further includes a driving motor for driving the turntable to rotate. A rack is arranged on the outer circumferential wall of the bottom of the turntable. A gear is fixedly arranged at the output end of the driving motor. The gear meshes with the rack.

[0021] A further setting of the present invention is that the demolding component includes a material conveying channel located below the blanking through groove and rotatably connected with the turntable. The material conveying channel is arranged in an S shape and air holes are evenly opened at the bottom of the material conveying channel. Collision baffle plates are arranged on both sides of the material conveying channel.

[0022] A further setting of the present invention is that the pressing component includes a pressing cylinder fixedly arranged on the workbench. A pressing plate is fixedly arranged on the piston rod of the pressing cylinder. A pressing block is arranged at the bottom of the pressing plate. The size of the pressing block is adapted to the size of the mold groove.

[0023] A further setting of the present invention is that the feeding component includes a bracket and a bidirectional motor. A mixing box is arranged on the bracket. A stirring motor is arranged on the top of the mixing box and a rotating rod is arranged inside. The stirring motor is used to drive the rotating rod to rotate. Mixing rods are arranged on the rotating rod. A blanking port is arranged at the bottom of the mixing box. A bidirectional lead screw is rotatably arranged on the bracket and a guiding rod is fixedly arranged. The bidirectional motor is used to drive the bidirectional lead screw to rotate. A sealing plate is slidably connected at the blanking port. One end of the sealing plate is threadedly connected to the bidirectional lead screw and the other end is slidably connected to the guiding rod.

[0024] A further setting of the present invention is that the waste material groove is of a trapezoidal structure and the two waist sides are tangent to the mold groove.

[0025] The beneficial effects of the present invention are as follows:

[0026] 1. In this application, the theaflavin and the fermentation pure liquid are mixed together in a buffer solution to form a freeze-dried powder containing SOD and then tableted. The phenolic hydroxyl groups and keto groups of theaflavin can form hydrogen bonds with the polar amino acid residues on the surface of SOD. The active center of Cu / Zn-SOD contains copper ions, and its empty orbitals can form coordination bonds with the ortho-phenolic hydroxyl groups or keto groups of theaflavin. The hydrogen bonds and coordination interactions together stabilize the structure of the complex. When in the intestine, the two work synergistically. Theaflavin can make up for the limitations of SOD against non-superoxide free radicals (such as hydroxyl radicals), broaden the antioxidant spectrum, and SOD can enhance the free radical scavenging efficiency of theaflavin and extend the antioxidant action time. At the same time, during the tableting process, theaflavin can also protect SOD. On the one hand, it actively absorbs light energy to erect a physical screen protection for SOD, and on the other hand, it scavenges the free radicals generated by photolysis, inhibits the metal-catalyzed oxidation reaction, erects a chemical defense for SOD, and reduces the photolysis loss of SOD during tableting.

[0027] 2. In this application, soy protein and wheat gliadin are selected and mixed as the coating layer of the tablet. Soy protein is rich in disulfide bonds and polar groups and has strong hydrophilicity. Mixing soy protein and wheat gliadin for coating can improve the hydrophilic performance of the coating layer to increase the loading capacity and loading efficiency of SOD. Wheat gliadin contains a large number of non-polar amino acids and has strong hydrophobicity. The polar groups of soy protein can also combine with the hydrophobic regions of gliadin through hydrogen bonds and hydrophobic interactions to form a more stable network structure, ensuring the stability of the coating layer, so that SOD is stably isolated inside the coating layer, achieving an effective and stable physical isolation from gastric acid. Experiments carried out in a simulated gastric acid environment show that after long-term soaking in gastric acid, the activity of SOD wrapped inside the coating layer is hardly lost (the activity remains above 95%), while the activity of the uncoated SOD is lost by more than 95% within the same time.

[0028] 3. In this application, an arc-shaped leveling plate is provided. The bottom of the arc-shaped leveling plate abuts against the turntable. As the turntable rotates, the leveling plate levels the SOD-containing powder in the mold groove, ensuring uniform distribution of the powder in the mold groove. During pressing, there is no need for excessive pressure to level, and only compaction is required to make the density and strength of each part of the tablet uniform, effectively avoiding the inactivation of SOD in the tablet due to excessive pressure. A waste material groove is provided between the mold grooves and the bottom of the waste material groove slopes downward. The opening of the leveling plate faces the feeding assembly. The waste material groove can not only collect the materials falling during the rotation of the turntable, but also scrape and collect the excess powder scraped by the leveling plate. The powder falls into the conical cylinder through the waste material groove, and the scraper inside the conical cylinder scrapes and collects the waste material, avoiding waste of the powder.

[0029] 4. In this application, a feeding component is provided to feed the SOD powder into the mold groove. As the turntable rotates, the mold groove filled with powder is moved below the leveling component. The leveling component levels the powder. Then the turntable rotates again, and the leveled mold groove is conveyed to the pressing component for pressing. After pressing, the turntable rotates again to drive the pressed sheet tablets to be conveyed to the blanking through groove and discharged and collected by the demolding component, realizing automatic and continuous blanking, leveling, pressing, and demolding and collection for the production of SOD tablets, with high processing efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0031] Figure 1 is the overall structural schematic diagram of a preparation device for SOD that resists pepsin degradation according to the present invention.

[0032] Figure 2 is the schematic diagram of the pressing component of a preparation device for SOD that resists pepsin degradation according to the present invention.

[0033] Figure 3 is the present invention Figure 2 partial enlarged view of A-A in.

[0034] Figure 4 is the schematic diagram of the feeding component of a preparation device for SOD that resists pepsin degradation according to the present invention.

[0035] Figure 5 is the present invention Figure 4 partial enlarged view of B-B in.

[0036] Figure 6 is the schematic diagram of the demolding component of a preparation device for SOD that resists pepsin degradation according to the present invention.

[0037] In the figure, 1 is a rotary tablet press; 2 is a workbench; 3 is a material guiding groove; 4 is a chassis; 5 is a turntable; 6 is a die groove; 7 is a waste material groove; 8 is a feeding assembly, 80 is a bracket, 81 is a bidirectional motor, 82 is a mixing box, 83 is a stirring motor, 84 is a rotating rod, 85 is a mixing rod, 86 is a blanking port, 87 is a bidirectional lead screw, 88 is a guide rod, 89 is a sealing plate; 9 is a leveling assembly, 90 is an L-shaped fixing bracket, 91 is a leveling plate; 10 is a pressing assembly, 100 is a pressing cylinder, 101 is a pressing plate, 102 is a pressing block; 11 is a demolding assembly, 110 is a material conveying channel, 111 is an air hole, 112 is a collision baffle; 12 is a blanking through groove, 13 is a conical cylinder, 14 is a rotating shaft, 15 is an auxiliary blanking assembly, 150 is a supporting cross bar, 151 is a scraping plate; 16 is a driving motor, 17 is a rack, 18 is a gear. Detailed implementation manners

[0038] The technical solutions of the present invention will be clearly and completely described below in conjunction with specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without making creative efforts belong to the scope of protection of the present invention.

[0039] Embodiment 1

[0040] The following steps are adopted to prepare the coated tablets 1 containing SOD:

[0041] S1. Powder preparation: Dissolve the SOD fermentation pure liquid with a purity greater than 95% in a phosphate buffer solution with a pH of 5.2 - 7.8, add a trehalose and cyclodextrin mixed solution with a volume ratio of 1:1 to the fermentation pure liquid, and then add theaflavin with a volume ratio of 1:1 to the fermentation pure liquid, stir for 30 min, filter and put it into a freeze dryer. First, cool it at a cooling rate of 1 - 3 °C / min to 40 °C to -60 °C and keep it for 2 - 4 h, then start the freeze dryer and freeze it for 12 - 48 h under the conditions of -50 °C to -30 °C and a high vacuum of 10 - 50 Pa to make freeze-dried powder.

[0042] S2. Drying: After S1 is completed, gradually raise the temperature to 0 °C - 25 °C and maintain the vacuum degree at 10 - 30 Pa for 2 - 10 h until the water content of the freeze-dried powder is reduced to less than 5%.

[0043] S3. Tableting: Mix the SOD freeze-dried powder evenly with the binder sorbitol, and the mass ratio of the binder to the SOD freeze-dried powder is 13 - 20:20. Then use a rotary tablet press to press it into tablets with a diameter of 3 - 8 mm;

[0044] S4. Coating solution preparation: Dissolve gliadin and soy protein in a mixed solution prepared with a volume ratio of water to ethanol of 1:1. Among them, the mass ratio of gliadin to soy protein is 0.5:1. Add glycerol accounting for 10% - 15% of the mass of gliadin, stir with a magnetic stirrer at room temperature and a speed of 500 rpm for 20 min, and sonicate with an ultrasonic device for 40 s;

[0045] S5. Coating: Place the tablets in a coating machine, set the air intake volume at 1500 - 1800 m 3 / h, the air inlet temperature at 35 - 40 °C, the pan rotation speed at 2 - 10 rpm, set the atomization rate at 2 - 5 mL / min, the fan surface pressure at 1.0 - 3.0 bar, and the spraying flow rate at 60 - 400 g / min, until the coating weight gain reaches 3% - 10% by mass fraction and the coating thickness is 20 - 50 μm, slowly dry at 40 °C for 1 h, and cool to obtain the coated tablets.

[0046] Example 2

[0047] The following steps are used to prepare coated tablets 2 containing SOD:

[0048] S1. Powder preparation: Dissolve the SOD fermentation pure liquid with a purity greater than 95% in a phosphate buffer solution with a pH of 5.2 - 7.8, add a trehalose and cyclodextrin mixed solution with a volume ratio of 1:1 to the fermentation pure liquid, and then add theaflavin with a volume ratio of 3:1 to the fermentation pure liquid, stir for 30 min, filter and put it into a freeze dryer. First, cool at a rate of 1 - 3 °C / min to 40 °C to -60 °C and maintain for 2 - 4 h, then start the freeze dryer and freeze at -50 °C to -30 °C and a high vacuum of 10 - 50 Pa for 12 - 48 h to make the freeze-dried powder.

[0049] S2. Drying: After S1 is completed, gradually raise the temperature to 0 °C - 25 °C, maintain the vacuum degree at 10 - 30 Pa for 2 - 10 h until the water content of the freeze-dried powder is reduced to less than 5%.

[0050] S3. Tabletting: Mix the SOD freeze-dried powder evenly with the binder sorbitol. The mass ratio of the binder to the SOD freeze-dried powder is 13 - 20:20, and then use a rotary tablet press to press into tablets with a diameter of 3 - 8 mm;

[0051] S4. Coating solution preparation: Dissolve gliadin and soy protein in a mixed solution prepared with a volume ratio of water to ethanol of 1:1. Among them, the mass ratio of gliadin to soy protein is 1:1. Add glycerol accounting for 10% - 15% of the mass of gliadin, stir with a magnetic stirrer at room temperature and a speed of 500 rpm for 20 min, and sonicate with an ultrasonic device for 40 s;

[0052] S5. Coating: Place the tablets in a coating machine, set the air intake volume at 1500 - 1800 m 3 / h, the air intake temperature at 35 - 40 °C, the pan rotation speed at 2 - 10 rpm, set the atomization rate at 2 - 5 mL / min, the fan surface pressure at 1.0 - 3.0 bar, the spraying flow rate at 60 - 400 g / min, until the coating weight gain reaches a mass fraction of 3% - 10% and the coating thickness is 20 - 50 μm, and then dry slowly at 40 °C for 1 h, and cool to obtain the coated tablets.

[0053] Example 3

[0054] The following steps are used to prepare coated tablets 3 containing SOD:

[0055] S1. Powder preparation: Dissolve the SOD fermentation pure liquid with a purity greater than 95% in a phosphate buffer solution with a pH of 5.2 - 7.8, add a trehalose and cyclodextrin mixed solution with a volume ratio of 1:1 to the fermentation pure liquid, and then add theaflavin with a volume ratio of 1:1 to the fermentation pure liquid, stir for 30 min, filter and put it into a freeze dryer. First, cool at a rate of 1 - 3 °C / min to 40 °C to -60 °C and keep it for 2 - 4 h, then start the freeze dryer and freeze at -50 °C to -30 °C and under a high vacuum of 10 - 50 Pa for 12 - 48 h to make the freeze-dried powder.

[0056] S2. Drying: After S1 is completed, gradually raise the temperature to 0 °C - 25 °C and maintain the vacuum degree at 10 - 30 Pa for 2 - 10 h until the water content of the freeze-dried powder is reduced to less than 5%.

[0057] S3. Tabletting: Mix the SOD freeze-dried powder evenly with the binder sorbitol, and the mass ratio of the binder to the SOD freeze-dried powder is 13 - 20:20. Then use a rotary tabletting machine to press into tablets with a diameter of 3 - 8 mm;

[0058] S4. Preparation of coating solution: Dissolve wheat gliadin and soy protein in a mixed solution prepared with a volume ratio of water:ethanol of 1:1. Among them, the mass ratio of wheat gliadin to soy protein is 0.5:1, add glycerol accounting for 10% - 15% of the mass of wheat gliadin, stir with a magnetic stirrer at room temperature and a speed of 500 rpm for 20 min, and ultrasonicate with an ultrasonic device for 40 s;

[0059] S5. Coating: Place the tablets in a coating machine, set the air intake volume at 1500 - 1800 m 3 / h, inlet air temperature at 35 - 40 °C, pan rotation speed at 2 - 10 rpm, set atomization rate at 2 - 5 mL / min, fan surface pressure at 1.0 - 3.0 bar, spraying flow rate at 60 - 400 g / min, until the coating weight gain reaches 3% - 10% by mass fraction and the coating thickness is 20 - 50 μm, slow drying at 40 °C for 1 h, and cooling to obtain the coated tablets.

[0060] Example 4

[0061] The following steps are used to prepare coated tablets 4 containing SOD:

[0062] Replace the gliadin in step S4 with zein, and the other steps are the same as in Example 1.

[0063] Comparative Example 1

[0064] The following steps are used to prepare coated tablets 5 containing SOD:

[0065] S1. Powder preparation: Dissolve the SOD fermentation pure liquid with a purity greater than 95% in a phosphate buffer solution with a pH of 5.2 - 7.8, add a trehalose and cyclodextrin mixed solution with a volume ratio of 1:1 to the fermentation pure liquid, then add an aqueous solution with a volume ratio of 1:1 to the fermentation pure liquid, stir for 30 min, filter and put it into a freeze dryer. First, cool it at a cooling rate of 1 - 3 °C / min to 40 °C to - 60 °C and hold for 2 - 4 h, then start the freeze dryer and freeze it for 12 - 48 h under the conditions of - 50 °C to - 30 °C and a high vacuum of 10 - 50 Pa to make the freeze-dried powder.

[0066] S2. Drying: After S1 is completed, gradually raise the temperature to 0 °C - 25 °C and maintain the vacuum degree at 10 - 30 Pa for 2 - 10 h until the water content of the freeze-dried powder is reduced to less than 5%.

[0067] S3. Tabletting: Mix the SOD freeze-dried powder evenly with the binder sorbitol, and the mass ratio of the binder to the SOD freeze-dried powder is 13 - 20:20. Then use a rotary tablet press to press it into tablets with a diameter of 3 - 8 mm;

[0068] S4. Coating solution preparation: Dissolve gliadin and soy protein in a mixed solution prepared with a volume ratio of water:ethanol of 1:1. Among them, the mass ratio of gliadin to soy protein is 0.5:1, add glycerol accounting for 10% - 15% of the mass of gliadin, stir with a magnetic stirrer at room temperature and a speed of 500 rpm for 20 min, and sonicate with an ultrasonic device for 40 s;

[0069] S5. Coating: Place the tablets in a coating machine, set the air inlet volume at 1500 - 1800 m 3 / h, inlet air temperature at 35 - 40 °C, pan rotation speed at 2 - 10 rpm, set atomization rate at 2 - 5 mL / min, fan surface pressure at 1.0 - 3.0 bar, spraying flow rate at 60 - 400 g / min, until the coating weight gain reaches 3% - 10% by mass fraction and the coating thickness is 20 - 50 μm, and then dry slowly at 40 °C for 1 h and cool to obtain the coated tablets.

[0070] Comparative Example 2

[0071] The following steps are used to prepare tablets 6 containing SOD:

[0072] S1. Powder preparation: Dissolve the SOD fermentation pure liquid with a purity greater than 95% in a phosphate buffer solution with a pH of 5.2 - 7.8, add a trehalose and cyclodextrin mixed solution with a volume ratio of 1:1 to the fermentation pure liquid, then add theaflavin with a volume ratio of 1 - 3:1 to the fermentation pure liquid, stir for 30 min, filter and put it into a freeze dryer. First, cool it at a cooling rate of 1 - 3 °C / min to 40 °C to - 60 °C and keep it for 2 - 4 h, then start the freeze dryer and freeze it for 12 - 48 h under the conditions of - 50 °C to - 30 °C and high vacuum of 10 - 50 Pa to make the freeze-dried powder.

[0073] S2. Drying: After S1 is completed, gradually raise the temperature to 0 °C - 25 °C and maintain the vacuum degree at 10 - 30 Pa for 2 - 10 h until the water content of the freeze-dried powder is reduced to less than 5%.

[0074] S3. Tabletting: Mix the SOD freeze-dried powder evenly with the binder sorbitol. The mass ratio of the binder to the SOD freeze-dried powder is 13 - 20:20. Then use a rotary tablet press to press it into tablets with a diameter of 3 - 8 mm to obtain the uncoated tablets 6.

[0075] Test method: Test the gastric acid resistance of the coated tablets 1 - 5 and tablets 6

[0076] Simulate the gastric acid environment: Prepare a hydrochloric acid solution with a pH of 1.2, which contains 0.3% (w / v) pepsin, as the simulated gastric acid environment.

[0077] Sample treatment: Take the coated tablets 1 - 5 and tablets 6 and add them to the simulated gastric acid solution, and incubate them in a constant temperature shaking incubator at 37 °C with a shaking speed of 100 rpm for 4 h.

[0078] Activity detection: After the incubation is completed, measure the activity of SOD in the solution through a specific SOD activity detection kit and compare it with the activity of the untreated SOD.

[0079] The results showed that after 4 hours of simulated gastric acid treatment, more than 95% of the SOD activity remained in coated tablets 1-4. Among them, coated tablet 2 had the best effect, retaining 99.3% of the SOD activity, while the SOD activity in coated tablet 5 remained at 85%, and the SOD activity in tablet 6 was lost by more than 98%. This indicates that the addition of theaflavin can increase the activity of the prepared SOD-coated tablets. At the same time, the coating layer formed by the mixture of soy protein and gliadin is more stable, which can protect SOD and prevent it from being degraded by pepsin in gastric acid.

[0080] Example 5

[0081] A preparation device for SOD resistant to pepsin degradation, which applies the above-mentioned preparation method for SOD resistant to pepsin degradation, as Figures 1-6 shown, includes a rotary tablet press 1. The rotary tablet press 1 includes a workbench 2. A feeding groove 3 is opened on the workbench 2. A chassis 4 is fixedly arranged in the feeding groove 3. A turntable 5 is rotatably connected above the chassis 4. Die grooves 6 are evenly arranged along the circumference of the turntable 5. There are at least four die grooves 6 and a waste material groove 7 is arranged between each die groove 6. The die groove 6 and the chassis 4 form a material storage space. Along the outer circumference of the turntable 5, a feeding component 8, a leveling component 9, a pressing component 10 and a demolding component 11 are arranged in sequence. A blanking through groove 12 is opened at the position of the chassis 4 where the demolding component 11 is located. The waste material groove 7 is arranged obliquely downward and a conical cylinder 13 is arranged below it. The top of the conical cylinder 13 is open. A rotating shaft 14 is fixedly arranged at the center of the turntable 5. The rotating shaft 14 penetrates through the conical cylinder 13 and an auxiliary blanking component 15 is arranged at the part of the rotating shaft 14 located inside the conical cylinder 13. Among them, the waste material groove 7 is of a trapezoidal structure, and the two waist sides are tangent to the die groove 6;

[0082] As Figure 6 shown, the auxiliary blanking component 15 includes a support cross bar 150 fixedly arranged on the rotating shaft 14. A scraping plate 151 is fixedly arranged at the end of the support cross bar 150 away from the rotating shaft 14. The bottom of the scraping plate 151 abuts against the inner wall of the conical cylinder 13 for scraping materials.

[0083] As Figure 6 shown, the leveling component 9 includes an L-shaped fixing frame 90 fixed on the workbench 2. A leveling plate 91 is arranged on the L-shaped fixing frame 90. The leveling plate 91 is arc-shaped and the opening faces the feeding component 8. The bottom of the leveling plate 91 is slidably connected with the turntable 5;

[0084] As Figure 3 、 Figure 6 shown, it also includes a driving motor 16 for driving the turntable 5 to rotate. A rack 17 is arranged on the outer circumferential wall at the bottom of the turntable 5. The output end of the driving motor 16 is fixedly provided with a gear 18, and the gear 18 meshes with the rack 17.

[0085] As Figure 6As shown, the demolding assembly 11 includes a material conveying channel 110 located below the blanking through groove 12 and rotatably connected to the turntable 5. The material conveying channel 110 is set in an S shape, and air holes 111 are evenly formed at the bottom of the material conveying channel 110. Collision baffles 112 are arranged on both sides of the material conveying channel 110.

[0086] As Figure 2 , Figure 3 As shown, the pressing assembly 10 includes a pressing cylinder 100 fixed on the workbench 2. A pressing plate 101 is fixed on the piston rod of the pressing cylinder 100. A pressing block 102 is arranged at the bottom of the pressing plate 101. The size of the pressing block 102 is adapted to the size of the mold groove 6.

[0087] As Figure 4 , Figure 5 , Figure 6 As shown, the feeding assembly 8 includes a bracket 80 and a bidirectional motor 81. A mixing box 82 is arranged on the bracket 80. A stirring motor 83 is arranged at the top of the mixing box 82, and a rotating rod 84 is arranged inside. The stirring motor 83 is used to drive the rotating rod 84 to rotate. Mixing rods 85 are arranged on the rotating rod 84. A blanking port 86 is arranged at the bottom of the mixing box 82. A bidirectional lead screw 87 is rotatably arranged on the bracket 80, and a guide rod 88 is fixed. The bidirectional motor 81 is used to drive the bidirectional lead screw 87 to rotate. A sealing plate 89 is slidably connected at the blanking port 86. One end of the sealing plate 89 is threadedly connected to the bidirectional lead screw 87, and the other end is slidably connected to the guide rod 88.

[0088] Its working principle is as follows: First, start the stirring motor 83 to drive the rotating rod 84 and the mixing rods 85 to rotate, so as to evenly mix the powder in the mixing box 82. Control the bidirectional motor 81 to drive the bidirectional lead screw 87 to rotate, thereby driving the sealing plate 89 to slide along the guide rod 88. When the sealing plate 89 moves away from the turntable 5, the blanking port 86 is gradually opened, and the blanking speed becomes faster. When the sealing plate 89 moves towards the turntable 5, the blanking port 86 is gradually closed, and the blanking speed becomes slower. The blanking speed can be flexibly adjusted to improve the tablet pressing efficiency. The powder falls from the falling port into the storage space formed by the lower mold groove 6 and the chassis 4.

[0089] When the stock space is discharged to a certain amount, the driving motor 16 is started to drive the gear 18 to rotate, thereby driving the turntable 5 to rotate. The turntable 5 rotates 90 degrees each time, driving the mold groove 6 filled with powder material to move to the leveling component 9. Then, the next mold groove 6 continues to be fed with material. After the feeding is completed, the turntable 5 rotates 90 degrees again. During the rotation process, the leveling plate 91 scrapes the powder material accumulated in the mold groove 6 flat, and scrapes the excess powder material and the powder material that falls on the turntable 5 during the rotation process into the waste material groove 7. After scraping, the mold groove 6 with the powder material leveled is located at the pressing component 10. At this time, the pressing cylinder 100 is started to drive the pressing plate 101 and the pressing block 102 to move downward to press the leveled powder material into a sheet shape. Then, the turntable 5 rotates 90 degrees again. At this time, the powder material in the next mold groove 6 is scraped flat, and the sheet-shaped finished product is moved to the blanking through groove 12 on the chassis 4 and automatically falls into the S-shaped material conveying channel 110 through the blanking through groove 12. And during the process of falling along the S-shaped material conveying channel 110, it continuously collides with the collision baffle 112, vibrating off the residual powder material on the tablet. The size of the air holes 111 is larger than the size of the powder material. The excess powder material falls through the air holes 111 and is collected. The tablets are collected through the discharge port of the material conveying channel 110. The empty mold groove 6 moves to the blanking port 86 again for feeding, and the processes of feeding, scraping flat, pressing, demolding and collecting are cycled in turn, realizing automatic tablet pressing and high processing efficiency;

[0090] When the powder material falling from the blanking port 86 during the rotation of the turntable 5 automatically falls into the waste material groove 7, the excess powder material generated by scraping flat and the powder material that falls on the turntable 5 during the rotation process are also scraped into the waste material groove 7. The bottom of the waste material groove 7 is inclined downward, and the powder material automatically falls into the opening of the conical cylinder 13. When the driving motor 16 is started to drive the turntable 5 to rotate, the rotating shaft 14 rotates accordingly, and then drives the support cross bar 150 and the scraping plate 151 to rotate. The scraping plate 151 abuts against the inner wall of the conical cylinder 13 to scrape the powder material falling on the inner wall of the conical cylinder 13, and finally falls from the blanking port 86 of the conical cylinder 13 to realize automatic collection and reuse, avoiding waste of powder material.

Claims

1. A preparation method of SOD resistant to pepsin degradation, characterized in that: It includes the following steps: S1. Powder preparation: Dissolve the SOD fermentation pure liquid with a purity greater than 95% in a buffer solution with a pH of 5.2 - 7.

8. Sequentially add a protective agent and theaflavin, stir for 30 min, filter, and then put it into a freeze dryer. First, reduce the temperature at a rate of 1 - 3 °C / min to 40 °C to -60 °C and maintain for 2 - 4 h. Then start the freeze dryer and freeze for 12 - 48 h under the conditions of -50 °C to -30 °C and a high vacuum of 10 - 50 Pa to make freeze-dried powder. S2. Drying: After S1 is completed, gradually increase the temperature to 0 °C - 25 °C and maintain the vacuum degree at 10 - 30 Pa for 2 - 10 h until the water content of the freeze-dried powder is reduced to less than 5%. S3. Tabletting: Mix the SOD freeze-dried powder evenly with a binder. The mass ratio of the binder to the SOD freeze-dried powder is 13 - 20:

20. Then use a rotary tablet press to press into tablets with a diameter of 3 - 8 mm. S4. Coating solution preparation: Dissolve gliadin and soy protein in a mixed solution made of water:ethanol with a volume ratio of 1:

1. Among them, the mass ratio of gliadin to soy protein is 0.5 - 1:

1. Add a plasticizer and stir with a magnetic stirrer at room temperature and a speed of 500 rpm for 20 min, and then use an ultrasonic device to ultrasonicate for 40 s. S5. Coating: Place the tablets in a coating machine, set the air intake volume at 1500 - 1800 m 3 / h, the inlet air temperature at 35 - 40 °C, the pan rotation speed at 2 - 10 rpm, set the atomization rate at 2 - 5 mL / min, the fan surface pressure at 1.0 - 3.0 bar, the spraying flow rate at 60 - 400 g / min, until the coating weight gain reaches a mass fraction of 3% - 10% and the coating thickness is 20 - 50 μm, slowly dry at 40 °C for 1 h, and cool to obtain the coated tablets.

2. The preparation method of SOD resistant to pepsin degradation according to claim 1, characterized in that: In the step S1, the specific protective agent is one or more of trehalose, cyclodextrin, and mannitol. The volume ratio of the protective agent to the fermentation pure liquid is 1:1, and the volume ratio of theaflavin to the fermentation pure liquid is 1 - 3:

1.

3. A preparation method of SOD resistant to pepsin degradation according to claim 1, characterized in that: In the step S2, the binder is one or more of sorbitol and maltodextrin.

4. A preparation method of SOD resistant to pepsin degradation according to claim 1, characterized in that: In the step S3, gliadin can be replaced by zein.

5. A preparation method of SOD resistant to pepsin degradation according to claim 1, characterized in that: In the step S3, the plasticizer is glycerol or polyethylene glycol 400, accounting for 10% - 15% of the mass of gliadin.

6. A preparation device for pepsin-resistant SOD, which applies the preparation method of pepsin-resistant SOD according to any one of claims 1-5, is characterized in that: It includes a rotary tablet press (1). The rotary tablet press (1) includes a workbench (2). A feeding groove (3) is opened on the workbench (2). A chassis (4) is fixedly arranged in the feeding groove (3). A turntable (5) is rotatably abutted above the chassis (4). Mold grooves (6) are evenly arranged along the circumference of the turntable (5). There are at least four mold grooves (6) and a waste material groove (7) is arranged between each mold groove (6). The mold groove (6) and the chassis (4) form a material storage space. Along the outer circumference of the turntable (5), a feeding assembly (8), a leveling assembly (9), a pressing assembly (10), and a demolding assembly (11) are sequentially arranged. A blanking through groove (12) is opened at the position of the chassis (4) where the demolding assembly (11) is located. The waste material groove (7) is arranged obliquely downward and a conical cylinder (13) is arranged below it. The top of the conical cylinder (13) is open. A rotating shaft (14) is fixedly arranged at the center of the turntable (5). The rotating shaft (14) penetrates through the conical cylinder (13) and an auxiliary blanking assembly (15) is arranged at the part of the rotating shaft (14) located inside the conical cylinder (13). The auxiliary blanking assembly (15) includes a support cross bar (150) fixed on the rotating shaft (14). A scraping plate (151) is fixed at one end of the support cross bar (150) away from the rotating shaft (14). The bottom of the scraping plate (151) is in contact with the inner wall of the conical cylinder (13) for scraping materials. The leveling assembly (9) includes an L-shaped fixing bracket (90) fixed on the workbench (2). A leveling plate (91) is provided on the L-shaped fixing bracket (90). The leveling plate (91) is arc-shaped and the opening faces the feeding assembly (8). The bottom of the leveling plate (91) is slidably connected to the turntable (5). It further includes a driving motor (16) for driving the turntable (5) to rotate. A rack (17) is provided on the outer circumferential wall of the bottom of the turntable (5). A gear (18) is fixed at the output end of the driving motor (16). The gear (18) meshes with the rack (17).

7. An SOD device resistant to pepsin degradation according to claim 6, characterized in that: The demolding assembly (11) includes a feeding channel (110) located below the blanking through slot (12) and rotatably connected to the turntable (5). The feeding channel (110) is arranged in an S shape and air holes (111) are evenly opened at the bottom of the feeding channel (110). Collision baffles (112) are provided on both sides of the feeding channel (110).

8. An SOD preparation device resistant to pepsin degradation according to claim 6, characterized in that: The pressing assembly (10) includes a pressing cylinder (100) fixed on the workbench (2). A pressing plate (101) is fixed on the piston rod of the pressing cylinder (100). A pressing block (102) is provided at the bottom of the pressing plate (101). The size of the pressing block (102) is adapted to the size of the mold groove (6).

9. An SOD preparation device resistant to pepsin degradation according to claim 6, characterized in that: The feeding assembly (8) includes a bracket (80) and a bidirectional motor (81). A mixing box (82) is provided on the bracket (80). A stirring motor (83) is provided at the top of the mixing box (82) and a rotating rod (84) is provided inside. The stirring motor (83) is used to drive the rotating rod (84) to rotate. Mixing rods (85) are provided on the rotating rod (84). A blanking port (86) is provided at the bottom of the mixing box (82). A bidirectional lead screw (87) is rotatably provided on the bracket (80) and a guide rod (88) is fixed. The bidirectional motor (81) is used to drive the bidirectional lead screw (87) to rotate. A sealing plate (89) is slidably connected at the blanking port (86). One end of the sealing plate (89) is threadedly connected to the bidirectional lead screw (87) and the other end is slidably connected to the guide rod (88).

10. The preparation device of SOD resistant to pepsin degradation according to claim 6, characterized in that: The waste material groove (7) is of a trapezoidal structure, and the two waist sides are tangent to the mold groove (6).

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