Traditional Chinese medicine purification device and use method

Abstract

The application provides a traditional Chinese medicine purification device, which comprises a first medicine liquid alcohol precipitation tank and a second medicine liquid alcohol precipitation tank, the first medicine liquid alcohol precipitation tank is connected with a first liquid storage tank, the second medicine liquid alcohol precipitation tank is connected with a second liquid storage tank, the first liquid storage tank and the second liquid storage tank are connected with a third liquid storage tank, a central transmission shaft is arranged in the first medicine liquid alcohol precipitation tank and the second medicine liquid alcohol precipitation tank, a spraying stirring structure and a medicine liquid extraction structure are arranged on the central transmission shaft, and a heating system is arranged on the spraying stirring structure. The use method of the traditional Chinese medicine purification device is characterized in that the spraying stirring structure uniformly sprays alcohol liquid in the medicine liquid alcohol precipitation tank and stirs the alcohol liquid at the same time, so that the alcohol liquid and the medicine liquid fully act on each other, the alcohol liquid is injected into the alcohol precipitation tank through the hole and is stirred at the same time, and the mixed liquid is sucked into the liquid storage tank through the hole, so that the structure is simple, the stirring is sufficient, the flow is controllable, and the filtration is sufficient.

Description

Technical Field

[0001] This invention belongs to the technical field of traditional Chinese medicine extraction, specifically a traditional Chinese medicine purification device and its usage method. Background Technology

[0002] Traditional Chinese medicine extraction refers to the use of extraction techniques to extract the effective components to the maximum extent, thereby improving the intrinsic quality and clinical therapeutic effect of traditional Chinese medicine preparations and maximizing the efficacy of traditional Chinese medicine.

[0003] The most commonly used separation and purification method in the production and manufacturing of traditional Chinese medicine is alcohol precipitation. The principle of alcohol precipitation is that the active ingredients in the extract of traditional Chinese medicine are soluble in ethanol solution, while impurities are insoluble in ethanol solution of a certain concentration. After adding ethanol to the extract, the active ingredients are transferred to the ethanol solution, while the impurities form a precipitate. After settling, solid-liquid separation is achieved, thus separating and purifying the active ingredients from the impurities. For example, the preparation method of Shenkang injection involves extracting the raw materials of traditional Chinese medicine according to the formula ratio and then subjecting them to multiple alcohol precipitation processes.

[0004] For example, the Chinese invention patent CN116036656A, published on May 2, 2023, discloses a traditional Chinese medicine alcohol precipitation device and control method, which belongs to the field of traditional Chinese medicine production technology. The device includes an alcohol precipitation tank, a settling tank, and a controller electrically connected to both. Both the precipitation tank and the settling tank are equipped with a liquid discharge component. A connecting pipe connects the two tanks, and a first supernatant discharge valve and an alcohol precipitation liquid discharge pump are installed on the connecting pipe. A first ultrasonic generator and a first temperature sensor are installed inside the precipitation tank. A medicinal liquid inlet and a first flocculant discharge valve are respectively installed at the top and bottom of the precipitation tank.

[0005] The aforementioned existing technologies effectively shorten alcohol precipitation time and improve production efficiency, but they do not fully extract the effective components in the drug solution or the liquid after precipitation, resulting in waste.

[0006] Therefore, how to effectively extract the active ingredients from the drug solution and the insufficient extraction of the settled liquid are urgent technical problems that need to be solved. Summary of the Invention

[0007] To address the shortcomings in the aforementioned background technology, this invention proposes a traditional Chinese medicine purification device and method, which solves the problems of insufficient extraction of effective components from medicinal liquids and insufficient extraction of the settled liquid.

[0008] The technical solution of this application is as follows:

[0009] A traditional Chinese medicine purification device includes a first medicinal liquid alcohol precipitation tank and a second medicinal liquid alcohol precipitation tank. The first medicinal liquid alcohol precipitation tank is connected to a first storage tank, and the second medicinal liquid alcohol precipitation tank is connected to a second storage tank. Both the first and second storage tanks are connected to a third storage tank. A central drive shaft is provided inside both the first and second medicinal liquid alcohol precipitation tanks. A spray stirring structure and a medicinal liquid extraction structure are provided on the central drive shaft. A heating system is provided on the spray stirring structure.

[0010] Furthermore, the spray mixing structure includes a mixing rod connected to the central drive shaft. The central drive shaft and the mixing rod are hollow structures with interconnected inner cavities. Several mixing rods are alternately arranged along the height direction of the central drive shaft. Several holes communicating with the inner cavity are provided on the circumferential surface of the mixing rod. Each hole is spirally distributed along the axis of the mixing rod, which increases the contact area between the outflowing alcohol liquid and the drug solution. The flow path of the alcohol liquid sprayed from each different spray hole is different, making the mixing more uniform and preventing clumping.

[0011] Furthermore, the bottom of the central drive shaft is provided with the second stirring rod. The second stirring rod is evenly distributed around the central drive shaft. The second stirring rod has holes facing downwards. The downward-facing holes can activate the clumps of particles deposited at the bottom of the tank, breaking them up and continuing to stir, so that the encapsulated active ingredients can continue to work and fully exert their effects.

[0012] Furthermore, the central drive shaft is connected to the feed pipe, which is equipped with a flow controller for controlling the supply of alcohol liquid. By monitoring, adjusting, and controlling the flow rate of the alcohol liquid through the flow controller, the flow rate of the fluid in the pipeline is effectively maintained constant, avoiding fluctuations and changes in flow rate, and ensuring stability and continuity.

[0013] Furthermore, the bottoms of the first and second drug solution alcohol precipitation tanks are conical. After alcohol precipitation is completed, impurities can accumulate at the conical bottom, which facilitates the drug solution extraction structure to absorb the liquid.

[0014] Furthermore, the drug extraction structure includes a third hole at the bottom of the central drive shaft, which is connected to a vacuum liquid aspirator. Filter screens are provided on the third hole, the first hole, and the second hole. The settled drug solution flows from the third hole, the first hole, and the second hole into the first storage tank.

[0015] Furthermore, a drive mechanism is provided on the central drive shaft, and a drive motor drives the central drive shaft to rotate.

[0016] Furthermore, temperature sensors are installed on the side walls of the first and second drug solution alcohol precipitation tanks.

[0017] Furthermore, a nanofiltration membrane is provided in the first or second liquid storage tank to filter components that need to be filtered out by the nanofiltration membrane.

[0018] A method of using a traditional Chinese medicine purification device includes the following steps:

[0019] S1: The different components of the drug solution are injected into the first drug solution alcohol precipitation tank from the inlet one and into the second drug solution alcohol precipitation tank from the inlet two.

[0020] S2: Temperature sensors installed on the side walls of the first and second drug solution alcohol precipitation tanks measure the temperature of the injected drug solution. If the temperature is higher than the set value, the stirring rod in the spray stirring structure will rotate to stir and cool the drug solution. If the temperature is lower than the set value, the heating system on the spray stirring structure will be turned on to heat the drug solution.

[0021] S3: When the temperature is suitable, add alcohol liquid to the first and second alcohol precipitation tanks. Under the control of the flow controller, the alcohol liquid flows into the central drive shaft through the feed pipe and then flows out from hole one. At the same time, the drive mechanism is turned on to rotate the central drive shaft. As the alcohol liquid flows in, stirring rod one fully stirs while spraying. The alcohol liquid flowing out from holes two and three of stirring rod two impacts the bottom of the first and second alcohol precipitation tanks, breaking up the clumps of alcohol liquid and allowing it to act again.

[0022] S4: After the stirring of the drug solution and alcohol liquid is completed, mixture A and mixture B are obtained respectively. The drive mechanism is turned off, and the mixture settles in the first drug solution alcohol precipitation tank and the second drug solution alcohol precipitation tank.

[0023] S5: After sedimentation, turn on the vacuum aspirator. Mixture A and mixture B are drawn back from holes one, two and three into the first and second storage tanks, respectively. Impurities are blocked by the filter screen and left in the first and second drug alcohol precipitation tanks.

[0024] S6: Mixture B passes through the nanofiltration membrane in the second storage tank and flows into the mixing tank together with mixture A for later use.

[0025] The specific beneficial effects of this invention include:

[0026] 1. Since the density of the drug solution and the alcohol liquid is different, the denser liquid tends to sink to the bottom of the device. The spray stirring structure in this invention runs through the entire drug solution alcohol precipitation tank. While spraying the alcohol liquid evenly and in a controlled manner in the drug solution alcohol precipitation tank, stirring is carried out to make the alcohol liquid and the drug solution fully interact.

[0027] 2. Because there are reaction factors between traditional Chinese medicine and alcohol liquids, the medicinal liquid has heat and is very easy to clump together when it comes into contact with cold ethanol. By scientifically controlling the temperature difference between the medicinal liquid and the alcohol liquid through the heating module, the medicinal liquid is fully precipitated by alcohol, saving resources and avoiding waste.

[0028] 3. The alcohol liquid is injected into the alcohol precipitation tank through the hole and stirred at the same time. The mixed liquid is then drawn back into the storage tank through the hole. The structure is simple, the stirring is thorough, the flow rate is controllable, and the filtration is thorough. 4. The liquid concentrated at the bottom of the tank can be drawn into the storage tank through hole three, which saves resources and costs and improves the extraction purity of alcohol precipitation.

[0029] 5. When the aqueous phase and alcohol phase are mixed, the insoluble components of the alcohol phase quickly precipitate to the bottom of the alcohol precipitation tank in the aqueous phase and agglomerate, adsorbing and encapsulating some of the active ingredients. Through the holes set at the bottom of the alcohol precipitation tank, the clumps precipitated at the bottom of the tank can be broken up and continue to react with the drug solution, so that the drug solution is fully precipitated with alcohol, the active ingredients are not easily lost, and resources and costs are saved.

[0030] 6. When multiple drug components need to be precipitated with alcohol, multiple alcohol precipitation tanks can be set up to carry out the precipitation simultaneously. The precipitated liquid is directly collected in a third storage tank for mixing and use, which improves efficiency and makes it convenient to use. Attached Figure Description

[0031] To more clearly illustrate the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0032] Figure 1 This is a schematic diagram of the present invention;

[0033] Figure 2 A schematic diagram of the central drive shaft, stirring rod 1, and stirring rod 2.

[0034] Figure 3 This is a schematic diagram of the fourth and fifth liquid storage tanks in this invention;

[0035] Figure 4 UPLC fingerprint of Shenkang injection;

[0036] Figure 5 The fingerprint spectrum is a control for Shenkang injection;

[0037] Figure 6 UPLC chromatogram of mixed control standards;

[0038] Figure 7UPLC chromatogram of the test sample for Shenkang injection;

[0039] Figure 8 Effects of Shenkang injection on histological damage and interstitial fibrosis in rats with renal failure (×200-fold);

[0040] Figure 9 Immunohistochemical results of kidneys after treatment with Shenkang injection for chronic renal failure (×200 times) (n=8, **P<0.01 compared with the control group; ##P<0.01 compared with the model group);

[0041] Figure 10 Effects of Shenkang injection on the expression of GRP78 and CHOP, important effector proteins of endoplasmic reticulum stress (n=8) (n=8, **P<0.01 compared with the control group; ##P<0.01 compared with the model group);

[0042] Figure 11 Results of the inhibition of fibrotic collagen deposition in the kidneys of rats with chronic renal failure by Shenkang injection (×200 times) (n=8, **P<0.01 compared with the control group; ##P<0.01 compared with the model group);

[0043] Figure 12 Effects of Shenkang injection on endoplasmic reticulum stress effector proteins in NRK-52E cells (n=6);

[0044] Figure 13 Effects of Shenkang injection on mitochondrial oxidative damage in NRK-52E cells (n=6, **P<0.01 compared with control group; ##P<0.01 compared with model group);

[0045] Figure 14 Effect of Shenkang injection on mitochondrial membrane potential (MMP) of NRK-52E cells (n=6, **P<0.01 compared with control group; ##P<0.01 compared with model group).

[0046] Explanation of icon numbers:

[0047] 1. First alcohol precipitation tank for the first drug solution; 2. Second alcohol precipitation tank for the second drug solution;

[0048] 3. Third storage tank; 4. Second storage tank;

[0049] 5. Third storage tank; 6. Central drive shaft; 7. Feed pipe;

[0050] 8. Liquid inlet 1; 9. Liquid inlet 2; 10. Drive mechanism;

[0051] 11. Stirring rod one; 12. Stirring rod two;

[0052] 13. Hole 1; 14. Hole 2; 15. Hole 3;

[0053] 21. Fourth storage tank; 22. Fifth storage tank. Detailed Implementation

[0054] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0055] A traditional Chinese medicine purification device, such as Figure 1 As shown, the device includes a first medicinal liquid alcohol precipitation tank 1 and a second medicinal liquid alcohol precipitation tank 2. The first medicinal liquid alcohol precipitation tank 1 is connected to a first storage tank 3, and the second medicinal liquid alcohol precipitation tank 2 is connected to a second storage tank 4. Both the first storage tank 3 and the second storage tank 4 are connected to a mixing tank 5. A central drive shaft 6 is installed inside both the first medicinal liquid alcohol precipitation tank 1 and the second medicinal liquid alcohol precipitation tank 2. A spray stirring structure and a medicinal liquid extraction structure are installed on the central drive shaft 6. A heating module is installed on the spray stirring structure. Because the medicinal liquid and the alcohol have different densities, the denser liquid tends to settle at the bottom of the device. The spray stirring structure in this invention runs through the entire medicinal liquid alcohol precipitation tank, uniformly and in controlled quantities spraying the alcohol while stirring, allowing the alcohol and medicinal liquid to fully react. Since there are reaction factors between the traditional Chinese medicine and the alcohol, the medicinal liquid possesses heat and is prone to agglomeration upon contact with cold ethanol. The heating module scientifically controls the temperature difference between the medicinal liquid and the alcohol, ensuring thorough alcohol precipitation of the medicinal liquid, saving resources and preventing waste.

[0056] Based on the above embodiments, as a preferred embodiment, the spray stirring structure includes a stirring rod 11 connected to the central drive shaft 6. The central drive shaft 6 and the stirring rod 11 are hollow structures with interconnected inner cavities. A plurality of stirring rods 11 are alternately arranged along the height direction of the central drive shaft 6. A plurality of holes 13 communicating with the inner cavity are provided on the circumferential surface of the stirring rod 11.

[0057] Specifically, such as Figure 2As shown, the central drive shaft 6 is located at the center of the first medicinal liquid alcohol precipitation tank 1 and the second medicinal liquid alcohol precipitation tank 2. Several rows of stirring rods 11 are arranged on the central drive shaft 6. Each row of stirring rods 11 has a different height and is evenly distributed along the height direction of the central drive shaft 6. This allows for all-round stirring of the medicinal liquid and alcohol liquid inside the first medicinal liquid alcohol precipitation tank 1. The alcohol liquid flows into the central drive shaft 6 from the inlet pipe at the top of the first medicinal liquid alcohol precipitation tank 1 and the second medicinal liquid alcohol precipitation tank 2, and then into the stirring rods 11. It then flows out from the hole 1 and comes into contact with the medicinal liquid in the first medicinal liquid alcohol precipitation tank 1 and the second medicinal liquid alcohol precipitation tank 2. The drive motor that drives the central drive shaft 6 to rotate is turned on, causing the stirring rods 11 to rotate in the alcohol precipitation tank. While rotating, the alcohol liquid slowly flows out from the hole 1 and comes into full contact with the medicinal liquid.

[0058] Preferably, each of the holes 13 is spirally distributed along the axis of the stirring rod 11, so that the contact area between the outflowing alcohol liquid and the medicine is larger. The flow path of the alcohol liquid sprayed from each different spray hole is different, so that the stirring is more uniform and clumping is prevented.

[0059] Based on the above embodiments, as a preferred embodiment, the bottom of the central drive shaft 6 is provided with a stirring rod 12, and several stirring rods 12 are evenly distributed around the central drive shaft 6. The holes 14 provided on the stirring rods 12 face downwards. Figure 2 As shown.

[0060] Specifically, the alcohol-precipitated particles cross-link and encapsulate with the medicinal liquid to form clumps, increasing their mass and depositing at the bottom of the container. The downward-facing pore 14 can activate the clumps of particles deposited at the bottom of the container, breaking them up and continuing to stir, allowing the encapsulated active ingredients to continue to work and fully exert their effects.

[0061] Based on the above embodiments, as a preferred embodiment, the central drive shaft 6 is connected to the feed pipe 7, and the feed pipe 7 is equipped with a flow controller for controlling the supply of alcohol liquid. By monitoring, adjusting, and controlling the flow rate of the alcohol liquid through the flow controller, the flow rate of the fluid in the pipeline is effectively maintained constant, avoiding fluctuations and changes in flow rate, and ensuring stability and continuity. The flow controller is usually used in conjunction with sensors, host computers, and other devices to achieve automated control and remote monitoring, reduce manual intervention, and improve work efficiency.

[0062] Based on the above embodiments, as a preferred embodiment, the bottom of the first drug solution alcohol precipitation tank 1 and the second drug solution alcohol precipitation tank 2 are conical. After alcohol precipitation is completed, impurities can accumulate at the conical bottom, which facilitates the drug solution extraction structure to absorb the liquid.

[0063] Based on the above embodiments, as a preferred embodiment, the drug extraction structure includes a third hole 15 at the bottom of the central drive shaft 6. The third hole 15 is connected to a liquid aspirator. Filter screens are provided on the third hole 15, the first hole 13, and the second hole 14. The settled drug solution flows from the third hole 15, the first hole 13, and the second hole 14 into the first storage tank 3. Impurities are blocked by the filter screen and remain in the first drug solution alcohol precipitation tank 1 and the second drug solution alcohol precipitation tank 2. The drug solution concentrated at the bottom of the tank can be drawn into the storage tank through the third hole 15, saving resources and costs and improving the extraction purity of alcohol precipitation.

[0064] Based on the above embodiments, as a preferred embodiment, a drive mechanism is provided on the central drive shaft 6, the drive mechanism including a drive motor, the drive motor driving the central drive shaft 6 to rotate.

[0065] Based on the above embodiments, as a preferred embodiment, temperature sensors are installed on the side walls of the first alcohol precipitation tank 1 and the second alcohol precipitation tank 2. The temperature sensors measure the temperature of the injected liquid medicine. If the temperature is higher than the set value, the stirring rod in the spray stirring structure is rotated to stir and cool the liquid medicine. If the temperature is lower than the set value, the heating system on the spray stirring structure is turned on to heat the liquid medicine.

[0066] Based on the above embodiments, as a preferred embodiment, the second liquid storage tank 4 is provided with a nanofiltration membrane, and the mixture B in the second liquid storage tank 4 contains components that need to be filtered out by the nanofiltration membrane.

[0067] Specifically, Shenkang Injection is a traditional Chinese medicine injection, whose components include... Rhubarb, Salvia miltiorrhiza, Safflower and Astragalus membranaceus This medication is primarily used to treat chronic renal failure, particularly suitable for those with dampness-heat and blood stasis syndrome. Shenkang Injection has the effects of relieving nausea and vomiting dampness, invigorating qi and promoting blood circulation, and clearing the bowels and eliminating dampness. In clinical applications, Shenkang Injection has shown significant therapeutic effects. Multiple studies have shown that it can effectively reduce serum urea nitrogen and creatinine levels while increasing hemoglobin levels. These effects help to alleviate the severity of chronic renal failure to some extent and reduce plasma viscosity and erythrocyte aggregation.

[0068] Specifically, the formula for preparing Shenkang Injection for treating chronic renal failure is as follows: 1500g rhubarb, 1500g salvia miltiorrhiza, 4500g astragalus membranaceus, and 1500g safflower.

[0069] The preparation method involves extracting the raw materials rhubarb and salvia miltiorrhiza according to the above formula proportions, followed by a first alcohol precipitation to remove tannins, a second alcohol precipitation followed by water precipitation, a third alcohol precipitation followed by a second water precipitation, and finally filtration. Then, extracting astragalus and safflower according to the above formula proportions, followed by a first alcohol precipitation followed by water precipitation, a second alcohol precipitation followed by water precipitation, and finally filtration. The two sets of liquids are then combined and subjected to ultrafiltration, fine filtration, dispensing, and sterilization. Alternatively, the raw materials rhubarb and salvia miltiorrhiza are extracted according to the above formula proportions, followed by a first alcohol precipitation to remove tannins, a second alcohol precipitation followed by water precipitation, a third alcohol precipitation followed by a second water precipitation, pyrogen removal, and sterilization. Then, extracting astragalus and safflower according to the above formula proportions, followed by a first alcohol precipitation followed by water precipitation, a second alcohol precipitation followed by water precipitation, pyrogen removal, and sterilization. Finally, the two sets of liquids are combined and subjected to ultrafiltration, fine filtration, dispensing, and sterilization.

[0070] Specifically, the effective components of safflower are quinone chalcones (safflower yellow and red pigments), flavonoids, spermidine, alkaloids, polyacetylenes, organic acids, etc. The nanofiltration membrane has a molecular weight cutoff of 200-600 Da, which can effectively retain flavonoid components and make the filtration of mixture B more refined.

[0071] Specifically, the upper part of the third liquid storage tank 5 is equipped with valve one connected to the first liquid storage tank 3 and valve two connected to the second liquid storage tank 4. After opening valve one and valve two, the mixed liquid A and mixed liquid B flow into the third liquid storage tank 5 and mix automatically, so that subsequent work can be carried out.

[0072] Specifically, when the mixture A in the first storage tank 3 needs to undergo multiple alcohol precipitations, only valve one is opened to allow mixture A to flow into the third storage tank 5, then it is drawn out and injected into the first drug alcohol precipitation tank 1 for repeated operation. Alternatively, the first storage tank 3 and the second storage tank 4 are each connected to different storage tanks, such as... Figure 3 As shown, the first storage tank 3 is connected to the fourth storage tank 21 and undergoes multiple alcohol precipitations separately. The second storage tank 4 is connected to the fifth storage tank 22 and undergoes multiple alcohol precipitations separately. When it is necessary to mix the two, the third storage tank 5 is used for mixing.

[0073] Specifically, when cleaning the first alcohol precipitation tank 1 and the second alcohol precipitation tank 2, pour clean water into the tanks to rinse them and remove impurities from the filter screen. The bottoms of both the first alcohol precipitation tank 1 and the second alcohol precipitation tank 2 can be opened to drain the impurities.

[0074] Specifically, when multiple drug components need to be precipitated with alcohol, multiple alcohol precipitation tanks can be set up to carry out the precipitation simultaneously. The precipitated liquid is directly collected in a third storage tank for mixing and use, which improves efficiency and makes it convenient to use.

[0075] A method of using a traditional Chinese medicine purification device includes the following steps:

[0076] S1: The different components of the drug solution are injected into the first drug solution alcohol precipitation tank 1 from the inlet 8 and into the second drug solution alcohol precipitation tank 2 from the inlet 9.

[0077] S2: Temperature sensors installed on the side walls of the first medicinal liquid alcohol precipitation tank 1 and the second medicinal liquid alcohol precipitation tank 2 measure the temperature of the injected medicinal liquid. If the temperature is higher than the set value, the stirring rod in the spray stirring structure will rotate to stir and cool the medicinal liquid. If the temperature is lower than the set value, the heating system on the spray stirring structure will be turned on to heat the medicinal liquid.

[0078] S3: When the temperature is suitable, add alcohol liquid to the first alcohol precipitation tank 1 and the second alcohol precipitation tank 2. Under the control of the flow controller, the alcohol liquid flows into the central drive shaft 6 through the feed pipe 7 and then flows out from the first hole 13. At the same time, the drive mechanism is turned on to rotate the central drive shaft 6. As the alcohol liquid flows in, the stirring rod 11 sprays and stirs thoroughly. The alcohol liquid flowing out from the second hole 14 and the third hole 15 of the stirring rod 12 impacts the bottom of the first alcohol precipitation tank 1 and the second alcohol precipitation tank 2, breaking up the clumps of alcohol liquid and allowing it to act again.

[0079] S4: After the stirring of the drug solution and alcohol liquid is completed, mixture A and mixture B are obtained respectively. The drive mechanism is turned off, and the mixture settles in the first drug solution alcohol precipitation tank 1 and the second drug solution alcohol precipitation tank 2.

[0080] S5: After sedimentation, turn on the vacuum suction device. Mixture A and mixture B are drawn back from hole 13, hole 24 and hole 35 into the first storage tank 3 and the second storage tank 4, respectively. Impurities are blocked by the filter screen and left into the first drug alcohol precipitation tank 1 and the second drug alcohol precipitation tank 2.

[0081] S6: Mixture B passes through the nanofiltration membrane in the second storage tank 4 and flows together with mixture A into the third storage tank 5 for later use. The relative density of the pharmaceutical solution is adjusted to 1.2 at 60°C using water for injection to obtain the injection solution.

[0082] Detection of the injection solution: chromatographic column was ACQUITY. HSS T3 (2.1 mm×100 mm, 1.8 μm); the mobile phase is acetonitrile (A)-0.1% formic acid water (B), with gradient elution: 0-5 min, 5% A; 5-10 min, 5%-10% A; 10-28 min, 10%-15% A; 28-43 min, 15%-25% A; 43-50 min, 25%-32% A; 50-62 min, 32%-50% A; 62-68 min, 50%-70% A; 68-75 min, 70%-100% A; 75-83 min, 100% A; 83-85 min, 100%-5% A; 85-90 min, 5% A. The injection volume is 10 μL; the flow rate is 0.2 mL·min-1; the column temperature is 40°C, and the detection wavelength is 254 nm.

[0083] The results are shown in Figure 4-7 , wherein Figure 4 is the UPLC fingerprint of Shenkang Injection; Figure 5 is the reference fingerprint of Shenkang Injection; Figure 6 is the UPLC chromatogram of mixed reference substances; Figure 7 is the UPLC chromatogram of the test sample of Shenkang Injection. It can be seen from the above figures that the quality of the Shenkang Injection of the present application is good.

[0084] Application Example

[0085] Experimental Animals

[0086] SPF-grade male SD rats (8 to 10 weeks old) were purchased from Henan Provincial Laboratory Animal Center, with license number: SCXK (Yu) 2022-0001, and certificate number: 4110062311. The rats were housed in a specific pathogen-free (SPF) animal room (12-hour light-dark cycle, temperature of 20-24°C and humidity of 40-70%), and provided with strictly controlled standard diet and distilled water. This experimental protocol was approved by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University (No.: 2023-KY-0543).

[0087] 1. Establishment of chronic renal failure model

[0088] After numbering 50 SD rats, 10 rats were randomly selected as the normal group, and the remaining 40 rats were administered with 200 mg·kg -1 dose of 2.5% adenine suspension by intragastric administration for 2 weeks, followed by 100 mg·kg -1 dose of 2.5% adenine suspension by intragastric administration for 1 week, once a day. After modeling, 3 rats were randomly selected from the normal group and the model group respectively, and blood was collected from the ophthalmic vein for detection of BUN and Scr (P800 automatic biochemical analyzer was used to detect the levels of BUN, Scr and UA in rat serum, and Advia 2400 biochemical analyzer was used to detect P in serum5+ Mg 2+ (Level). After sacrifice, kidney tissue was harvested to observe kidney size and perform HE staining for pathological observation. As shown in Table 1, BUN and Scr were significantly higher than in the normal group, and the presence of fibrosis and inflammatory cell infiltration in the pathological observation indicated successful model establishment.

[0089] Blood biochemistry test results showed that the model group had levels of Bun, Scr, UA, and P. 5+ and Mg 2+ All indicators were significantly elevated (P<0.01), indicating that the kidneys had already experienced dysfunction and were unable to maintain homeostasis, leading to the accumulation of toxic metabolites and disturbances in water, electrolyte, and acid-base balance. The low- and high-dose groups of Shenkang injection significantly reduced these indicators (P<0.01), indicating that Shenkang injection has an effect on improving kidney function. In contrast, the positive control group only improved the relevant indicators to a certain extent, but the effect was not significant. This is because the positive control group mainly acts as an antioxidant and cannot comprehensively improve kidney function.

[0090] Table 1 Blood biochemical indicators of different experimental groups

[0091]

[0092] Note: a P<0.01, compared with the control group; b P<0.05, compared with the model group; c P<0.01, compared with the model group

[0093] Forty rats that successfully developed the model were then randomly divided into a model group, a low-dose Shenkang group, a high-dose Shenkang group, and a positive control group, with 10 rats in each group. The daily treatment regimen was as follows: the low-dose Shenkang group received a tail vein injection of 3 mL / kg Shenkang injection, the high-dose Shenkang group received a tail vein injection of 6 mL / kg Shenkang injection, the positive control group received an intraperitoneal injection of the reactive oxygen species (ROS) inhibitor NAC (5 mg / kg), and the normal group and the model group received an intravenous injection of an equal volume of physiological saline. This treatment was administered once daily for 14 consecutive days. After the treatment, the rats were sacrificed, and their kidneys and serum were collected and stored at -80°C or fixed in 10% neutral formalin and then embedded in paraffin for further analysis.

[0094] 2. Cell culture and in vitro experiments

[0095] The rat renal tubular epithelial cell line NRK-52E was cultured in DMEM medium + 10% FBS at 37℃, 95% air and 5% CO2. The cultured NRK-52E cells were randomly divided into 4 groups: (1) Control group: NRK-52E cells were starved in DMEM containing 1% FBS for 24h, and then cultured in DMEM containing 10% FBS for 24h. (2) TBHP group: After starvation, NRK-52E cells were cultured in DMEM medium supplemented with 10% fetal bovine serum and 150μmol / L TBHP (tert-butanol peroxide, a commonly used oxidant for constructing cellular oxidative damage) for 24h to construct a renal cell oxidative stress damage model. (3) TBHP+NAC group: During starvation treatment, 5mmol / L NAC was added to the culture medium, and then 150μmol / L TBHP was added for 24h treatment. (4) TBHP+SK (animal serum containing Shenkang) group: The drug-containing serum of the low-dose Shenkang injection group was added to the culture medium during starvation treatment, followed by incubation with 150 μmol / L TBHP for 24 h. The preparation method of the drug-containing serum was as follows: normal rats were injected with Shenkang injection (3 mL / kg) via the tail vein twice a day for 3 consecutive days. One hour after the last administration, blood was collected from the orbital cavity and centrifuged at low temperature (parameters set to 3000 r / min, 3 min). The drug-containing serum was obtained after centrifugation and inactivated by heating in a 56℃ water bath for 30 min. The drug-containing serum was made into lyophilized powder. 0.5 mg of the lyophilized powder was added to 10 mL of blank serum, which is equivalent to a final concentration of 50 μg / mL.

[0096] ① Kidney histopathological evaluation: HE staining was used to evaluate changes in kidney histological morphology (kidney tissue from each group of rats was fixed in 10% neutral formalin buffer and embedded in paraffin. Sections (5μm) were stained with hematoxylin and eosin (HE). Images of each slide were acquired using an optical microscope and evaluated blinded by a pathologist.)

[0097] In the control group, glomeruli were evenly distributed in the cortex, with uniform cell number and matrix within the glomeruli. No obvious abnormalities were observed in the renal tubules, with neat and regular brush borders and no obvious abnormalities in the medulla. The connective tissue between the urinary tubules was the renal interstitium, with no obvious interstitial proliferation. No obvious inflammatory changes were observed. Compared with the control group, in the model group rats, the kidney tissue showed extensive renal tubular dilation and flattened epithelial cells; a large number of renal tubular epithelial cells showed hydropic degeneration with loose and pale cytoplasm; diffuse connective tissue proliferation was observed in the interstitium between the renal tubules, accompanied by extensive lymphocyte infiltration; a large number of renal tubules atrophied, with narrowing or disappearance of the lumen; a small number of protein casts were observed; urate crystals were observed in a large number of renal tubules; and a small number of necrotic cells were observed in a small number of renal tubules. Compared with the model group, the renal tissue of the Shenkang and positive drug control groups showed a small amount of renal tubular dilation; a small amount of hydropic degeneration of renal tubular epithelial cells; a small amount of connective tissue hyperplasia between renal tubules, accompanied by local lymphocyte infiltration; and a small amount of urate crystals within the renal tubules. The pathological condition was significantly improved. Figure 8 .

[0098] ② Immunohistochemistry: Paraffin-embedded sections (5 μm) were dewaxed and rehydrated using a gradient of ethanol and water. Antigens were activated by microwave in citrate buffer (pH 6.0), and then placed in H2O2 solution for 15 min to block endogenous peroxidase. The slides were then incubated overnight at 4°C with primary antibodies of KIM-1 (1:300), GRP78 (1:600), CHOP (1:100), and α-SMA (1:100), respectively. After incubation with the corresponding secondary antibodies for 1 h at room temperature, the slides were stained with 3,3'-Diaminobenzidine tetrahydrochloride (DAB), followed by hematoxylin counterstaining to identify cell nuclei. Images of the immunohistochemically stained slides were obtained using an optical microscope.

[0099] KIM-1 is a biomarker for proximal tubular injury in the kidney. This study investigated the protective effect of Shenkang injection on kidney injury by detecting KIM-1 expression in kidney tissue using immunohistochemistry. Results showed a significant increase in KIM-1-positive tubular cells in the model group. Both the Shenkang group and the positive control group significantly reduced the number of KIM-1-positive tubular cells. These results indicate that Shenkang injection can effectively protect renal filtration function, maintain the integrity of kidney morphology, and alleviate kidney injury. Figure 9 .

[0100] Endoplasmic reticulum (ER) stress and oxidative stress are interrelated and play important roles in the pathogenesis of various human diseases. To investigate whether the protective effect of Shenkang injection on renal injury is related to the regulation of ER stress, this study used immunohistochemical staining and Western blotting to detect the expression of important ER stress effector proteins GRP78 and CHOP. Both immunohistochemical staining and Western blotting showed that, compared with the control group, the expression of GRP78 and CHOP was significantly increased in the model group, while the expression of GRP78 and CHOP was significantly reduced after treatment with Shenkang injection and positive control drugs. The results indicate that Shenkang injection can effectively inhibit excessive ER stress in pathological renal conditions, thereby exerting a renal protective effect. Figure 9 and Figure 10 .

[0101] NRK-52E cells were cultured in 24-well slides, fixed with 4% paraformaldehyde, and stained with immunofluorescence for 10 minutes. The slides were blocked with 5% BSA and 0.5% Triton X-100 at room temperature for 2 hours, then incubated overnight at 4°C with primary antibody against GRP78 (1:200). After washing with PBS, the slides were incubated with the corresponding fluorescent secondary antibody, and cell nuclei were identified using DAPI. Images were captured using a fluorescence microscope. It was observed that Shenkang injection could inhibit the expression of KIM-1, GRP78, and CHOP, thereby suppressing endoplasmic reticulum over-stress and exerting a renal protective effect.

[0102] ③ Western blot analysis: In in vivo sample analysis, kidney tissue (including cortex and medulla) was homogenized, and total protein was extracted using RIPA lysis buffer. In in vitro sample analysis, cultured NRK-52E cells were collected, and total protein was extracted using cell lysis buffer. Equal volumes of total protein were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene fluoride (PVDF) membrane. Endoplasmic reticulum stress was detected using primary antibodies against GRP78 (1:1000), anti-CHOP (1:1000), and anti-ATF4 (1:1000). Using anti-GAPDH antibody as a control, protein bands were visualized using a Bio-Rad ChemiDoc XRS+ chemiluminescence imaging system after reaction with primary antibody and horseradish peroxidase (HRP)-labeled secondary antibody.

[0103] To observe collagen deposition in renal interstitial fibrosis, this study performed immunohistochemical analysis on type I collagen, the most abundant collagen in the ECM. The results showed that in the control group, Collagen I was mainly distributed in the vessel wall, while in the model group, Collagen I expression was significant, mainly distributed in the renal tubular basement membrane. The levels of type I collagen in the renal tubular basement membrane were significantly reduced in both the renal health group and the positive control group.

[0104] To further verify the protective effect of Shenkang injection on renal interstitial fibrosis, this study performed α-SMA immunohistochemical staining to detect myofibroblast proliferation. The results showed that in the control group rats, α-SMA was mainly expressed in vascular smooth muscle cells. In the model group rats, α-SMA was significantly increased in the kidneys, mainly located on the renal tubular basement membrane, and renal tubules atrophied; conversely, Shenkang injection and the positive control significantly inhibited myofibroblast proliferation. In summary, these results indicate that Shenkang injection can effectively inhibit the proliferation of myofibroblasts, reduce ECM deposition, and thus inhibit the formation of renal interstitial fibrosis. Figure 11 .

[0105] ④ Detection of reactive oxygen species (ROS): The ROS production in NRK-52E cells was detected using a reactive oxygen species detection kit. NRK-52E cells were incubated with a reactive oxygen species fluorescent probe (DCFH-DA) at 37°C for 20 min, washed three times in PBS, and the cell nuclei were identified by DAPI co-staining. Images were captured using a fluorescence microscope.

[0106] To further verify the relationship between the protective effect of Shenkang injection and excessive endoplasmic reticulum stress, this study used TBHP-induced NRK-52E cells as a cell model of oxidative stress injury. Immunofluorescence staining showed that TBHP treatment significantly increased the expression of GRP78 in NRK-52E cells, while NAC or Shenkang serum treatment effectively inhibited GRP78 production. Furthermore, Western blot analysis showed that TBHP treatment significantly increased the production of important endoplasmic reticulum stress effector proteins such as GRP78, CHOP, and ATF-4 in NRK-52E cells, while NAC and Shenkang serum treatment significantly reduced the production of these proteins. These results indicate that the protective effect of Shenkang injection against kidney injury is related to the inhibition of excessive endoplasmic reticulum stress. Figure 12 It can be seen that Shenkang injection inhibits excessive endoplasmic reticulum stress in NRK-52E cells.

[0107] ⑤ Mitochondrial membrane potential (MMP) measurement: The MMP of NRK-52E cells was detected using the JC-1 mitochondrial membrane potential detection kit. NRK-52E cells were incubated with JC-1 solution at 37°C for 20 minutes, and then washed with 1×JC-1 staining buffer. Images were captured using a fluorescence microscope. The red-green fluorescence ratio was analyzed using ImageJ software.

[0108] Endoplasmic reticulum stress and oxidative stress are related, and both play important roles in the pathogenesis of various human diseases, such as kidney disease. Studies have reported that excessive ROS production is associated with the pathogenesis of oxidative stress and triggers mitochondrial damage. In this in vitro oxidative stress-induced cell model, TBHP treatment increased ROS production in NRK-52E cells, while NAC and renal serum significantly reduced excessive ROS production. (See [link to study]). Figure 13 .

[0109] JC-1Assay was used to detect the mitochondrial membrane potential (MMP) of NRK-52E cells. The results showed that the relative red-to-green fluorescence intensity in TBHP-treated NRK-52E cells was significantly lower than that in control NRK-52E cells. However, the relative red-to-green fluorescence intensity in the TBHP+NAC group was higher than that in the TBHP group. Furthermore, renal serum showed a similar effect to NAC in maintaining the MMP of NRK-52E cells. These data indicate that renal serum can alleviate oxidative damage by reducing excessive ROS production and maintaining MMP. Figure 14 .

[0110] All aspects not detailed in this invention are conventional technical means known to those skilled in the art.

[0111] The above content shows and describes the basic principles, main features, and beneficial effects of the present invention. The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine purification device, characterized in that: The system includes a first alcohol precipitation tank (1) and a second alcohol precipitation tank (2). The first alcohol precipitation tank (1) is connected to a first storage tank (3), and the second alcohol precipitation tank (2) is connected to a second storage tank (4). Both the first storage tank (3) and the second storage tank (4) are connected to a third storage tank (5). A central drive shaft (6) is installed inside both the first alcohol precipitation tank (1) and the second alcohol precipitation tank (2). The central drive shaft (6) is equipped with a spray stirring structure and a drug extraction structure. The spray stirring structure is equipped with a heating system to prevent the drug solution from agglomerating after encountering cold ethanol, scientifically controlling the temperature difference between the drug solution and the alcohol liquid, and ensuring sufficient alcohol precipitation of the drug solution. The spray stirring structure includes... A stirring rod 1 (11) is connected to the central drive shaft (6). The central drive shaft (6) and the stirring rod 1 (11) are hollow structures with interconnected inner cavities. The circumferential surface of the stirring rod 1 (11) is provided with several holes 1 (13) that communicate with the inner cavity. Each hole 1 (13) is spirally distributed along the axis of the stirring rod 1 (11). A stirring rod 2 (12) is provided at the bottom of the central drive shaft (6). The hole 2 (14) provided on the stirring rod 2 (12) is oriented downwards. The drug extraction structure includes a hole 3 (15) provided at the bottom of the central drive shaft (6). The settled drug solution flows from hole 3 (15), hole 1 (13), and hole 2 (14) into the first storage tank (3). The method of using the device includes the following steps: S1: The different components of the drug solution are injected into the first drug solution alcohol precipitation tank (1) through the inlet one (8) and into the second drug solution alcohol precipitation tank (2) through the inlet two (9); S2: Temperature sensors installed on the side walls of the first medicinal liquid alcohol precipitation tank (1) and the second medicinal liquid alcohol precipitation tank (2) measure the temperature of the injected medicinal liquid. If the temperature is higher than the set value, the stirring rod in the spray stirring structure will rotate to stir and cool the medicinal liquid. If the temperature is lower than the set value, the heating system on the spray stirring structure will be turned on to heat the medicinal liquid. S3: When the temperature is suitable, add alcohol liquid to the first alcohol precipitation tank (1) and the second alcohol precipitation tank (2). The amount of alcohol liquid delivered is controlled by the flow controller and flows into the central drive shaft (6) through the feed pipe (7) and then flows out from the first hole (13). At the same time, the drive mechanism is turned on to make the central drive shaft (6) rotate. As the alcohol liquid flows in, the first stirring rod (11) sprays and stirs fully. The alcohol liquid flowing out from the second hole (14) and the third hole (15) of the second stirring rod (12) impacts the bottom of the first alcohol precipitation tank (1) and the second alcohol precipitation tank (2), breaking up the clumps of alcohol liquid and acting again. S4: After the mixing of the drug solution and alcohol liquid is completed, the first drug solution alcohol precipitation tank (1) and the second drug solution alcohol precipitation tank (2) respectively obtain mixture A and mixture B. The drive mechanism is turned off, and sedimentation takes place in the first drug solution alcohol precipitation tank (1) and the second drug solution alcohol precipitation tank (2). S5: After sedimentation, turn on the vacuum suction device. Mixed liquid A and mixed liquid B are drawn back from hole one (13), hole two (14), and hole three (15) into the first storage tank (3) and the second storage tank (4), respectively. Impurities are blocked by the filter screen and left in the first drug alcohol precipitation tank (1) and the second drug alcohol precipitation tank (2). S6: Mixture B passes through the nanofiltration membrane in the second storage tank (4) and flows together with mixture A into the third storage tank (5) for later use.

2. The traditional Chinese medicine purifying device according to claim 1, characterized in that: Several stirring rods (11) are alternately arranged along the height direction of the central drive shaft (6).

3. The traditional Chinese medicine purifying device according to claim 2, characterized in that: The stirring rods (12) are evenly distributed around the central drive shaft (6).

4. The traditional Chinese medicine purification device according to any one of claims 1-3, characterized in that: The central drive shaft (6) is connected to the feed pipe (7), and the feed pipe (7) is equipped with a flow controller for controlling the supply of alcohol liquid.

5. The traditional Chinese medicine purification apparatus according to any one of claims 1-3, characterized in that: The bottoms of the first drug solution alcohol precipitation tank (1) and the second drug solution alcohol precipitation tank (2) are conical.

6. The traditional Chinese medicine purification apparatus according to any one of claims 1-3, characterized in that: The third hole (15) is connected to the vacuum liquid aspirator, and filters are provided on the third hole (15), the first hole (13), and the second hole (14).

7. The traditional Chinese medicine purification apparatus according to any one of claims 1-3, characterized in that: A drive mechanism (10) is provided on the central drive shaft (6).

8. The traditional Chinese medicine purification apparatus according to any one of claims 1-3, characterized in that: Temperature sensors are installed on the side walls of the first drug solution alcohol precipitation tank (1) and the second drug solution alcohol precipitation tank (2).

9. The traditional Chinese medicine purification apparatus according to any one of claims 1-3, characterized in that: The first liquid storage tank (3) is equipped with a nanofiltration membrane.

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