Extraction method of active substances of parnassia palustris and application of active substances in anti-tumor drugs

Abstract

The invention discloses an extraction method of active substances of parnassia palustris and application of the active substances to antitumor drugs, and relates to the technical field of oncotherapy, and the extraction method comprises the following steps: putting dried parnassia palustris into a crushing device for crushing; extracting the crushed parnassia palustris, saturated saline solution and 2-octanol in an empty bottle, adding a magnetic stirrer during extraction, sealing in an SPME operation table, and extracting a 1-tetradecanol extracting solution from the parnassia palustris after extraction; the application has the technical advantages that the active substance 1-tetradecanol is extracted from parnassia palustris, and the 1-tetradecanol reduces the energy supply of tumor cells by inhibiting the energy metabolism pathway of the tumor cells, so that the metabolic state of the tumor cells is changed to a certain extent, and the tumor cells can be effectively inhibited. The method has important significance in understanding the influence mechanism of drugs on the growth and survival of tumor cells; and a crushing structure composed of a cyclone separator and a centrifugal crusher is arranged, so that efficient crushing of parnassia palustris is realized, and stripping of wet air in parnassia palustris particles is realized.

Description

Technical Field

[0001] The present application relates to the technical field of tumor treatment, and in particular to a method for extracting active substances from Herba Eupatorii and its application in anti-tumor drugs. Background Art

[0002] Throughout humanity's long struggle against disease, malignant tumors have always been one of the most formidable threats to life and health. With the rapid advancement of modern medicine, anti-tumor drug research has evolved from traditional cytotoxic chemotherapy into a new era of precision and personalized treatment. Targeted therapies specifically inhibit the molecular pathways of tumor cells, while immunotherapies activate the body's own immune system to fight cancer cells. These breakthroughs have significantly prolonged patient survival. However, challenges such as tumor heterogeneity, drug resistance mechanisms, and treatment toxicity remain, urgently requiring the scientific and medical communities to work together to overcome them. Against this backdrop, Traditional Chinese Medicine (TCM) has re-entered the global anti-tumor research landscape due to its unique advantages: natural ingredients, stable effects, and minimal toxicity. TCM not only maintains the naturalness and bioactivity of its ingredients, but also, due to its ease of absorption and metabolism by the body, avoids the cumulative toxicity caused by long-term chemical residues and reduces the risk of drug resistance.

[0003] Tumors, as pathological neoplasms formed by the abnormal proliferation of the body under the influence of tumorigenic factors, have become a highly challenging malignant disease in contemporary medicine. Against the backdrop of accelerated industrialization, a surge in environmental pollutants, and increasing social pressure, their incidence has shown a significant upward trend. Although the main clinical treatments for malignant tumors, such as cytotoxic chemotherapy, radiotherapy, and radical surgery, can curb the progression of the disease, they generally have problems such as high treatment costs and severe immune damage. Some patients even suffer from secondary organ dysfunction due to treatment toxicity. Therefore, how to establish an anti-tumor Chinese medicine system based on the advantages of active substances in herbal medicines has become a technical problem to be solved by those skilled in the art, as well as how to achieve large-scale production of active substances and solve the problem of regular, non-stop automatic cleaning of the screen in the pulverizing equipment used for batch production. Summary of the Invention

[0004] This device provides a method for extracting active substances from Prunus mume and its application in anti-tumor drugs. The specific implementation methods are as follows:

[0005] This device provides a method for extracting active substances from euphorbia pulcherrima, comprising the following steps:

[0006] Step 100: Place the dried safflower grass into a crushing device for crushing;

[0007] Step 200: Extract the crushed Herba Cistanches, saturated saline, and 2-octanol in an empty bottle, add a magnetic stirrer during the extraction, and seal the bottle in an SPME operating table. After the extraction, extract 1-tetradecanol extract from the Herba Cistanches.

[0008] Preferably, each extraction operation in step 200 is pre-equilibrated at 70° C. for 10 minutes, and the extraction operation includes solid phase microextraction, gas chromatography and mass spectrometry analysis.

[0009] Preferably, in the extraction operation of step 200, the solid phase microextraction conditions are as follows: the aging extraction head is placed in the SPME operating table of the GERSTEL three-in-one autosampler for headspace adsorption for 20 minutes, and then the extraction head is inserted into the GC injection port for adsorption for 5 minutes, and the heating program is started at the same time.

[0010] Preferably, in the extraction operation of step 200, the gas chromatography conditions are: inlet temperature 200°C, non-split injection, injection time 1 min, total flow rate 20 mL / min, purge flow rate 3 mL / min, and carrier gas He; programmed temperature conditions: initial chromatographic column temperature 30°C, retained for 1 min, increased to 92°C at a rate of 14°C / min, retained for 2 min, increased to 200°C at a rate of 5°C / min, retained for 7 min, increased to 220°C at a rate of 6°C / min, and maintained for 8 min.

[0011] Preferably, in the extraction operation of step 200, the mass spectrometry conditions are: ion source temperature 200°C: GC-MS interface temperature 200°C: solvent delay time 5 min; mass scan range (m / z) 35-500, full scan mode.

[0012] The crushing device used in the method for extracting active substances from the herb includes:

[0013] A centrifugal grinder and a cyclone separator connected to the centrifugal grinder through a connecting pipe, wherein a switching assembly is provided at the bottom of the cyclone separator;

[0014] The switching assembly includes a disc structure composed of several receiving bins arranged along the circumference. The outer part of the disc structure is connected to a discharge cover for rotation. Each receiving bin is open to the outside and has an inclined screen built in. The inclined screen for screening the material is regularly replaced by the rotation of the disc structure.

[0015] Based on the above technical solution, the crushing structure is composed of a cyclone separator and a centrifugal crusher. After the safflower enters the crushing structure, the centrifugal crusher generates centrifugal force by rotating at high speed, efficiently crushing the safflower, and quickly breaking it into small particles. During the crushing process, the particles collide and rub against the internal components of the equipment, which fully destroys the structure of the safflower. The cyclone separator uses the centrifugal force field generated by high-speed rotation to form a rotating airflow within the separator between the moist air and the crushed safflower particles. During this process, the moist air, due to its lower density, is closer to the center of the rotating airflow, while the safflower particles are thrown toward the inner wall of the separator under the action of centrifugal force, thereby achieving separation between the two.

[0016] Preferably, a coarse screen box is provided between the equal-diameter cylinder and the conical cylinder of the cyclone separator, and a coarse screen is installed in the coarse screen box through a vibration assembly, and the vibration assembly is connected to a rotatable disc structure through a downward linkage through a push rod, and the disc structure is integrated with a square frame acting on the push rod, and the coarse screen is vibrated and prevented from blocking by the rotation of the disc structure.

[0017] Preferably, the top of the discharge hood is vertically connected to the conical cylinder through a material guide opening, a return material interface is opened on one side of the discharge hood, and a fracture is opened at the bottom, through which the bottom end inclined screen is cleaned and maintained, and the large particles of plum blossom grass in the receiving bin are radially discharged through the return material interface, and the small particles of plum blossom grass pass through the inclined screen and are axially discharged from the receiving bin.

[0018] Preferably, square frames are coaxially provided at both ends of the disc structure, one side of the square frame is laterally provided with a discharge port connected to the receiving bin, and the other side of the square frame is connected to the first motor.

[0019] Preferably, it also includes a fine screening box composed of a box body, the disc structure is arranged inside the box body, and the end of the box body is rotatably connected to the square frame through a driven disc; the side of the box body is provided with a lateral opening at a position close to the discharge port, and a cleaning hole connected to the fracture is opened at the bottom of the box body.

[0020] Based on the above technical solution, by setting up a switching component, the circumferential regular replacement of the inclined screen is achieved; a coarse screen is added at the front end of the inclined screen to disperse the adhered plum grass particles, making subsequent screening more accurate; the rotation of the switching component can indirectly drive the coarse screen and the vibration component to vibrate, and simultaneously realize the regular vibration cleaning of the coarse screen, avoiding the blockage of the coarse screen and ensuring the screening efficiency.

[0021] Preferably, the vibration assembly includes a movable seat, which is slidably connected to the first vertical guide groove on the coarse screen box through a guide column; the movable seat is vertically divided into three layers: a top plate, an intermediate plate and a bottom plate, and the top plate and the bottom plate are respectively connected to the conical cylinder and the return material interface through elastic rubber sleeves; the coarse screen is installed between the intermediate plate and the bottom plate, and a fastener is provided on the intermediate plate, and a driving structure that is driven by the fastener is provided between the top plate and the intermediate plate;

[0022] Preferably, the coarse screen is horizontally inserted into the guide seat, and its end abuts against the positioning back plate, and the positioning back plate and the coarse screen box are respectively provided with a second vertical guide groove and a pad for bottom support after the coarse screen is inserted, and the vertical movable height of the bottom plate is lower than the insertion height of the coarse screen;

[0023] Preferably, the fastener comprises a coaxially arranged transmission gear and a flipping claw, the bottom of the transmission gear is engaged with a tooth plate slidably connected to the bottom plate, and the end of the tooth plate is downwardly provided with a wedge block acting on the bottom of the coarse screen;

[0024] Preferably, a gear ring is provided between the top plate and the middle plate, the gear ring is engaged with the output end of the second motor, the gear ring is also engaged with any transmission gear on the same side, and a linkage member with reverse synchronous translation is provided between the two gear plates.

[0025] Based on the above technical solution, by arranging fasteners on the vibration assembly, the coarse screen can be manually replaced while the switching assembly is operating normally and without stopping the machine.

[0026] The application of the active substance of Herba Citri Reticulatae in anti-tumor drugs, wherein the active substance of Herba Citri Reticulatae is 1-tetradecanol.

[0027] Preferably, 1-tetradecanol is used in anti-glioma drugs.

[0028] In summary, this application has the following beneficial technical effects:

[0029] 1. This invention extracts the active substance 1-tetradecanol from Herba Eupatorii. 1-tetradecanol inhibits the energy metabolism pathway of tumor cells, reducing the energy supply of tumor cells, thereby changing the metabolic state of tumor cells to a certain extent. This is of great significance for understanding the mechanism by which drugs affect the growth and survival of tumor cells.

[0030] 2. The present invention achieves efficient pulverization of the safflower grass by providing a pulverization structure composed of a cyclone separator and a centrifugal pulverizer. It also removes the moist air from the safflower grass particles, thereby improving the overall pulverization accuracy and dryness of the material.

[0031] 3. The present invention uses a switching assembly to allow the inclined screen to be regularly replaced circumferentially. A coarse screen is added in front of the inclined screen to disperse adhering sedge particles, making the screening of sedge particles more accurate. The rotation of the switching assembly indirectly drives the coarse screen and the vibration assembly to vibrate, thereby achieving regular vibration cleaning of the coarse screen.

[0032] 4. The present invention has a simple structure. By arranging fasteners on the vibration assembly, the coarse screen can be manually replaced without stopping the machine or switching the assembly during normal operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is a result diagram of the TU686 plate cloning validation of 1-tetradecanol in the present invention;

[0034] Figure 2 This is a result diagram of the TU212 plate cloning validation of 1-tetradecanol in the present invention;

[0035] Figure 3 This is a result diagram of the CNE2 plate cloning validation of 1-tetradecanol in the present invention;

[0036] Figure 4 This is a diagram showing the results of cloning verification of 1-tetradecanol using NPC / HK-1 plates in the present invention;

[0037] Figure 5 This is a diagram showing the invasion and metastasis results of CNE2 in the present invention;

[0038] Figure 6 This is a graph showing the invasion and metastasis results of NPC / HK-1 in the present invention;

[0039] Figure 7 This is a graph showing the invasion and metastasis results of TU212 in the present invention;

[0040] Figure 8 This is the result diagram of U251 invasion and metastasis in the present invention;

[0041] Figure 9 The change in total ATP content when exploring the ability to inhibit cell proliferation in this invention Figure 1 ;

[0042] Figure 10 The change in total ATP content when exploring the ability to inhibit cell proliferation in this invention Figure 2 ;

[0043] Figure 11 This is a diagram showing the Seahorse test results of U87MG in the present invention;

[0044] Figure 12 This is an ECAR graph showing the effect of U87MG under 1-1-tetradecanol treatment in the present invention;

[0045] Figure 13 This is an OCAR diagram showing the effect of U87MG under 1-1-tetradecanol treatment in the present invention;

[0046] Figure 14 This is a diagram of the Seahorse experiment results of U251 in the present invention;

[0047] Figure 15 This is the ECAR graph of the effect of U251 under 1-1-tetradecanol treatment in the present invention;

[0048] Figure 16 This is the OCAR diagram of the effect of U251 under 1-1-tetradecanol treatment in the present invention;

[0049] Figure 17 This is a schematic diagram of the structure of the present invention Figure 1 ;

[0050] Figure 18 This is a schematic diagram of the structure of the present invention Figure 2 ;

[0051] Figure 19 is a cross-sectional view of the structure of the present invention;

[0052] Figure 20 It is a schematic structural diagram of the switching component and the vibration component in the present invention;

[0053] Figure 21 It is a schematic diagram of the exploded structure of the discharge cover and the switching assembly in the present invention;

[0054] Figure 22 is a cross-sectional view of the switching assembly structure of the present invention;

[0055] Figure 23 This is a schematic diagram of the structure of the switching component after it moves relative to the discharge cover in the present invention;

[0056] Figure 24 is a cross-sectional view of the structure of the switching component and the vibration component in the present invention;

[0057] Figure 25 is a cross-sectional view of the coarse screen box and the vibrating assembly structure of the present invention;

[0058] Figure 26 Schematic diagram of the cross-section explosion of the coarse screen and the vibration assembly structure of the present invention;

[0059] Figure 27 It is a schematic diagram of the explosion structure of the coarse screen and the vibration assembly in the present invention;

[0060] Figure 28 It is a partially enlarged structural diagram of the vibration component in the present invention;

[0061] Figure 29 This is a partially enlarged schematic diagram of the fastener in the present invention. Figure 1 ;

[0062] Figure 30 This is a partially enlarged schematic diagram of the fastener in the present invention. Figure 2 ;

[0063] Figure 31 It is a schematic diagram of the explosion structure of the vibration component in the present invention.

[0064] Description of reference numerals:

[0065] 1. Fine screen box, 2. Cyclone separator, 3. Coarse screen box, 4. Connecting pipe, 5. Discharge cover, 6. Centrifugal crusher, 7. Switching component, 8. Coarse screen, 9. Vibration component,

[0066] 101, cleaning hole, 102, driven disk, 103, box, 104, side opening, 201, equal diameter cylinder, 202, exhaust pipe, 203, tapered cylinder, 301, positioning back plate, 302, guide seat, 303, first vertical guide groove, 3011, second vertical guide groove, 501, return material interface, 502, guide opening, 503, fracture, 601, crusher body, 602, feed port, 701, square frame, 702, inclined screen, 703, first motor, 704, receiving bin, 705, discharge port, 706, limit guide Groove, 801, positioning rod, 802, plate body, 803, grid structure, 901, push rod, 902, abutment plate, 903, movable seat, 904, elastic rubber sleeve, 905, fastener, 906, linkage part, 907, guide column, 908, gear ring, 909, second motor, 9031, top plate, 9032, middle plate, 9033, bottom plate, 9034, horizontal guide groove, 9051, transmission gear, 9052, flip claw, 9053, gear plate, 9054, wedge block, 9061, connecting rod, 9062, elastic slide plate. DETAILED DESCRIPTION

[0067] The specific implementation of the present invention is described below with reference to the accompanying drawings and embodiments:

[0068] It should be noted that the structures, proportions, sizes, etc. illustrated in the drawings of this specification are only used to match the contents disclosed in the specification for people familiar with this technology to understand and read, and are not used to limit the conditions under which the present invention can be implemented. Any structural modification, change in proportional relationship or adjustment of size should still fall within the scope of the technical content disclosed in the present invention without affecting the efficacy and purpose that can be achieved by the present invention.

[0069] At the same time, the terms such as "upper", "lower", "left", "right", "middle" and "one" quoted in this specification are only for the convenience of description and are not used to limit the scope of implementation of the present invention. Changes or adjustments to their relative relationships should be regarded as the scope of implementation of the present invention without substantially changing the technical content.

[0070] The following is combined with Figure 1-31 This application is described in further detail.

[0071] The examples of the present application disclose a method for extracting active substances from Herba Eupatorii and its application in anti-tumor drugs.

[0072] Example 1

[0073] Reference Figures 1 to 16 This embodiment discloses a method for extracting active substances from Prunus mume, and the specific steps include:

[0074] Step 100: Place the dried safflower grass into a crushing device for crushing;

[0075] Step 200: Extract the crushed herba syringae, saturated saline, and 2-octanol in an empty bottle, add a magnetic stirrer during the extraction, and seal the bottle in an SPME operating table. After the extraction, extract 1-tetradecanol from the herba syringae;

[0076] Step 300: placing the 1-tetradecanol extract in a centrifuge and centrifuging it. After the centrifugation, the Herba Eupatorii extract generates a supernatant, and impurities are precipitated at the bottom of the centrifuge tube.

[0077] Step 400: Pour the centrifuged 1-tetradecanol extracts into Buchner funnels for suction filtration to obtain pure 1-tetradecanol extracts;

[0078] Step 500: Turn on the refrigerator, install the eggplant-shaped bottle containing the 1-tetradecanol extract on the rotary evaporator, and perform water bath rotary evaporation. Evaporation and concentration are carried out until the extract is nearly dry, and then the rotary evaporation operation can be stopped.

[0079] Step 600: Place the eggplant-shaped bottle after rotary evaporation concentration into a vacuum drying oven, set the drying temperature and time, remove residual solvent and moisture, and improve the purity of the 1-tetradecanol extract.

[0080] Step 700: Determine the concentration range of the 1-tetradecanol extract. Calculate the amount of DMSO solution required based on the concentration of the extract and the required sample volume. Use a micropipette to add the DMSO solution to dissolve the extract and prepare a 1-tetradecanol sample solution. When weighing, weigh the crushed safflower herb, the empty eggplant-shaped bottle, and the eggplant-shaped bottle after sample preparation in sequence to calculate the extraction rate.

[0081] In the above method steps, the centrifugation time of the centrifuge tube in step 300 is set to 5 minutes, and the speed of the centrifuge is set to 3000-5000 r / min. In step 500, the solvent extract is introduced into the eggplant-shaped bottle before the water bath and positioned with a clamp. The height and rotation speed of the eggplant-shaped bottle are adjusted during the water bath, and the water bath temperature is set; in step 200, each extraction operation is pre-equilibrated at 70°C for 10 minutes, and the extraction operation includes solid phase microextraction, gas chromatography and mass spectrometry analysis; the solid phase microextraction conditions are: use an aged extraction head to enter the SPME operating table of the GERSTEL three-in-one automatic sampler for headspace adsorption for 20 minutes, then insert the extraction head into the GC injection port for adsorption for 5 minutes, and at the same time start The gas chromatography conditions were as follows: injection port temperature 200°C, splitless injection, injection time 1 min, total flow rate 20 mL / min, purge flow rate 3 mL / min, and carrier gas He; programmed temperature conditions: initial column temperature 30°C, retained for 1 min, increased to 92°C at a rate of 14°C / min, retained for 2 min, increased to 200°C at a rate of 5°C / min, retained for 7 min, increased to 220°C at a rate of 6°C / min, and maintained for 8 min; mass spectrometry conditions were as follows: ion source temperature 200°C: GC-MS interface temperature 200°C: solvent delay time 5 min; mass scanning range (m / z) 35-500, full scan mode.

[0082] Example 2

[0083] Reference Figures 17 to 24 Based on the above embodiment, this embodiment also discloses a pulverizing device used in the method for extracting active substances from plum blossom grass, a centrifugal grinder 6 and a cyclone separator 2 connected to the centrifugal grinder 6 through a connecting pipe 4, a switching component 7 is provided at the bottom of the cyclone separator 2, and the switching component 7 includes a disc structure composed of several receiving bins 704 arranged along the circumference, and the outside of the disc structure is rotatably connected to a discharge cover 5, each receiving bin 704 is open to the outside, and has an inclined screen 702 built in. The inclined screen 702 for screening is regularly replaced by rotating the disc structure. In this structure, the centrifugal grinder 6 has a conventional grinder body 601, and the grinder body 601 has an axially arranged feed port 602.

[0084] A coarse screen box 3 is provided between the equal-diameter cylinder 201 and the conical cylinder 203 of the cyclone separator 2, and a coarse screen 8 is installed in the coarse screen box 3 through a vibration assembly 9, and the vibration assembly 9 is connected to a rotatable disc structure downward through a push rod 901. The disc structure is integrated with a square frame 701 acting on the push rod 901, and the coarse screen 8 is prevented from vibrating and blocking by the rotation of the disc structure. An exhaust pipe 202 is installed on the top of the equal-diameter cylinder 201. In this structure, the square frame 701 is provided with a limiting guide groove 706 along its circumference, and the end of the push rod 901 is provided with an abutment plate 902 that is slidably connected to the limiting guide groove 706, thereby improving the stability of the vertical sliding of the push rod 901.

[0085] The top of the discharge hood 5 is vertically connected to the conical cylinder 203 through the material guide opening 502. A return material interface 501 is provided on one side of the discharge hood 5, and a fracture 503 is provided at the bottom. The bottom end inclined screen 702 is cleaned and maintained through the fracture 503, and the large particles of plum blossom grass in the receiving bin 704 are radially discharged through the return material interface 501, and the small particles of plum blossom grass pass through the inclined screen 702 and are axially discharged from the receiving bin 704.

[0086] Square frames 701 are coaxially provided at both ends of the disc structure. One side of the square frame 701 is laterally provided with a discharge port 705 connected to the receiving bin 704, and the other side of the square frame 701 is connected to the first motor 703. In this structure, the disc structure is arranged inside the box body 103, and the end of the box body 103 is rotatably connected to the square frame 701 through the driven disc 102; the side of the box body 103 is provided with a lateral opening 104 at a position close to the discharge port 705, and the bottom of the box body 103 is provided with a cleaning hole 101 connected to the fracture 503. A brush can be installed at the position of the cleaning hole 101 in this structure. During the flipping of the disc structure, the brush automatically cleans the inclined screen 702, and the inclined screen 702 can also be disassembled and assembled from the bottom; the return material interface 501 can be directly incorporated into the feed port 602 through conventional feeding equipment, so that the uncrushed plum blossom grass can be re-introduced into the feed port 602 for multiple crushing.

[0087] Example 3

[0088] Reference Figures 17 to 31 Based on the above embodiments, this embodiment also discloses a crushing device used in the method for extracting active substances from plum blossom grass. The vibration component 9 includes a movable seat 903, which is slidably connected to the first vertical guide groove 303 on the coarse screen box 3 through a guide column 907. The movable seat 903 is vertically divided into three layers: a top plate 9031, an intermediate plate 9032 and a bottom plate 9033. The top plate 9031 and the bottom plate 9033 are respectively connected to the conical cylinder 203 and the return material interface 501 through an elastic rubber sleeve 904; and a fastener 905 is provided on the intermediate plate 9032, and a driving structure that is transmitted to the fastener 905 is provided between the top plate 9031 and the intermediate plate 9032. In this structure, the coarse screen 8 is installed between the intermediate plate 9032 and the bottom plate 9033. The distance between the intermediate plate 9032 and the bottom plate 9033 is greater than the thickness of the coarse screen 8, and a curtain is provided at the end of the guide seat 302 for sealing the interior after the coarse screen 8 is inserted.

[0089] Guide seats 302 and positioning back plates 301 are provided on both sides of the coarse screen box 3. The coarse screen 8 is horizontally inserted into the guide seat 302, and its end is against the positioning back plate 301. The positioning back plate 301 and the coarse screen box 3 are respectively provided with a second vertical guide groove 3011 and a pad for bottom support after the coarse screen 8 is inserted, and the vertical movable height of the bottom plate 9033 is lower than the insertion height of the coarse screen 8; the screen of the coarse screen 8 in this structure can also be further expanded, and its flow channel can be designed as a dehumidification material. At the same time, with the help of regular vibration of the coarse screen 8, the particles can be prevented from adhering to the dehumidification material.

[0090] The fastener 905 includes a coaxially arranged transmission gear 9051 and a flipping claw 9052. The bottom of the transmission gear 9051 is engaged with a tooth plate 9053 that is slidably connected to the bottom plate 9033, and a wedge block 9054 is provided at the end of the tooth plate 9053 to act on the bottom of the coarse screen 8.

[0091] A gear ring 908 is provided between the top plate 9031 and the middle plate 9032. The gear ring 908 is engaged with the output end of the second motor 909. The gear ring 908 is also engaged with any transmission gear 9051 on the same side. A linkage member 906 for reverse synchronous translation is provided between the two gear plates 9053. In this structure, the linkage member 906 includes an elastic slide 9062 and a horizontal guide groove 9034 provided on the top plate 9031. The elastic slide 9062 and the horizontal guide groove 9034 are connected. They are perpendicular to each other, and the end of the tooth plate 9053 is provided with an endpoint that is slidably inserted into the horizontal guide groove 9034. The elastic slide 9062 is a conventional spring, slider and slide groove structure, and the endpoints of the tooth plate 9053 are connected to both sides of the slider through connecting rods 9061. When the tooth plate 9053 on one side is translated along the direction of the horizontal guide groove 9034, the spring is compressed and the slider is driven, and in this state, the tooth plate 9053 on the other side also translates synchronously in the opposite direction along the direction of the horizontal guide groove 9034.

[0092] The specific implementation process is as follows: the plum blossom grass is fed into the feed port 602 of the centrifugal crusher 6, and the plum blossom grass crushed by the cyclone separator 2 enters the cyclone separator 2 through the connecting pipe 4. After the gas-solid separation is completed in the cyclone separator 2, the crushed plum blossom grass falls downward into the coarse screen 8. The mesh size of the coarse screen 8 is larger than that of the inclined screen 702, which is used to block large particles of impurities. The mesh size of the coarse screen 8 can also be set to cater to the lateral opening shape of the plum blossom grass particles.

[0093] The safflower grass particles fall vertically from the bottom into the receiving bin 704; the safflower grass particles with appropriate apertures pass through the inclined screen 702 and are directly discharged from the discharge port 705; the safflower grass particles with large apertures are periodically driven by the first motor 703 to rotate the disc structure, so that the safflower grass particles in the receiving bin 704 are discharged laterally from the return interface 501 and re-enter the centrifugal crusher 6;

[0094] When the first motor 703 drives the disc structure to rotate, the square frame 701 rotates synchronously, acting on the coarse screen 8, the movable seat 903 and the push rod 901 at the same time, realizing the vibration and anti-blocking of the coarse screen 8; when the disc structure completes the rotation, the elastic rubber sleeve 904 forces the coarse screen 8 and the movable seat 903 to return to their original position;

[0095] When the coarse screen 8 is replaced, the coarse screen 8 is pushed in from the guide seat 302 and is placed horizontally on the second vertical guide groove 3011 and the pad; the disc structure and the centrifugal crusher 6 operate normally, and the movable seat 903 does not touch the coarse screen 8 during vertical vibration; without stopping the machine, the coarse screen 8 is manually pulled out from the guide seat 302 during the time interval when the disc structure stops rotating; after replacing the coarse screen 8, the second motor 909 installed at the top plate 9031 is started to drive the gear ring 908 to rotate, and the two sides of the gear ring 908 are respectively transmitted to the fasteners 905, and the pair of fasteners 905 on the same side move in opposite directions with the help of the linkage member 906; the hook-shaped flipping claw 9052 flips down and acts on the bottom of the coarse screen 8, the tooth plates 9053 on both sides move inward, and the inclined surface of the wedge block 9054 presses on the bottom of the coarse screen 8, so that the coarse screen 8 is locked on the movable seat 903 for synchronous vibration.

[0096] Example 4

[0097] Reference Figures 26 to 28 Based on the above embodiments, this embodiment also discloses a crushing device used in the method for extracting active substances from kelp. The coarse screen 8 includes a plate body 802, a grid structure 803 is installed in the middle of the plate body 802, and a positioning rod 801 is provided on the side of the plate body 802. The positioning rod 801 is inserted into the second vertical guide groove 3011 on the positioning back plate 301.

[0098] A double-layer grid structure 803 can also be installed in the middle of the plate body 802. The double-layer grid structure 803 is staggered, and a conventional elastic closed structure is set between the two. A trigger head is provided on the top of the elastic closed structure. The disc structure and the square frame 701 rotate. When the plate body 802 is forced to move up to a fixed point with the help of the vibration component 9, the trigger head abuts against the coarse screen box 3 to close the double-layer grid structure 803, so that the plum blossom grass material on the upper layer of the double-layer grid structure 803 continues to be discharged when the switching component 7 is triggered.

[0099] Example 5

[0100] Reference Figures 1 to 16 This embodiment also discloses the application of the active substance of Herba Eupatorii in anti-tumor drugs. The active substance of Herba Eupatorii is 1-tetradecanol, and the application of 1-tetradecanol in anti-glioma drugs.

[0101] Example 6

[0102] Reference Figures 1 to 16Based on Example 5, this example also discloses a method for verifying the effect of the active substance of Herba Eupatorii after its application in anti-tumor drugs; in order to explore the inhibitory effect of 1-tetradecanol on tumors, it was respectively applied to two types of brain glioma cells, U87MG and U251, and the MTT (MTT: 3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide) cell viability assay was used to detect the effect of 1-tetradecanol on the viability of brain glioma cells.

[0103] The principle of the inhibition assay is that succinate dehydrogenase, present in the mitochondria of living cells, can be reduced by MTT molecules into water-insoluble, blue-purple crystalline formazan, which then deposits within the cells. Dead cells lack this enzyme activity and therefore cannot undergo this reduction reaction. By adding dimethyl sulfoxide (DMSO) to dissolve the intracellular formazan crystals, the absorbance (OD) at a wavelength of 490 nm can be measured, which indirectly reflects the number of viable cells. Within a certain cell population range, the amount of formazan crystals formed is proportional to the number of viable cells. The inhibition assay involves plating 5,000 cells per well (100 μl) of U87MG / U251 cells in the logarithmic growth phase in a 96-well plate. After overnight attachment, the cells are treated with the drug at varying concentrations and durations. After the target treatment time, add the prepared MTT solution (6ul per well), place in a cell culture incubator and continue to culture for 2-4h, discard the cell culture medium, add 100ul DMSO liquid to each well, and measure its absorbance at a wavelength of 490nm using a microplate reader.

[0104] When exploring the effect of 1-tetradecanol on the proliferation and clone formation ability of single cells, a plate cloning experiment was used for verification. The specific experimental method was as follows: logarithmically growing cells were collected and counted, and 1000 cells (2 ml culture medium) were plated per well of a 6-well plate. After the cells adhered (about 1 day later), drug treatment (by changing the medium) was performed for 10-14 days, and the clone formation was observed. The cell culture medium was discarded, paraformaldehyde was added for fixation for 15 minutes, and then crystal violet was used for staining for 15 minutes. The excess crystal violet stain was washed off, and the cells were dried. The size and number of clones were observed. Figures 1 to 4 , represent experimental verification using TU686 plate, TU212 plate clones, NPC / HK-1 plate clones and CNE2 plate clones, respectively. Part A is a diagram of clone formation in the blank control group, Part B is a diagram of clone formation in the 1-1-tetradecanol-treated group, Part C is a microscopic image of monoclonal clones in the blank control group, Part D is a microscopic image of monoclonal clones in the 1-1-tetradecanol-treated group, and Part E is a bar graph of the number of clones in the blank control on the 1-1-tetradecanol plate.

[0105] When investigating the effect of 1-1-tetradecanol on the invasion and migration ability of tumor cells, the specific experimental method was to collect logarithmically growing cells, count them, plate 10-50w cells per well in the upper chamber of the Transwell cell plate chamber, and add different treatment factors to the lower chamber; divide them into control group (cultured with DMEM high glucose medium), 1-1-tetradecanol treatment group (treated with 2μM drug concentration for 48h); combine Figures 5 to 8 , respectively, are the results of CNE2 invasion and metastasis, NPC / HK-1 invasion and metastasis, TU212 invasion and metastasis, and U251 invasion and metastasis. The left side of each figure is the Transwell experiment microscopic result figure, and the right side is the Transwell experiment counting result bar chart; by comparing the invasion and metastasis abilities of different cells after treatment with 1-1-tetradecanol, the research results show that the invasion and metastasis abilities of cells treated with 1-1-tetradecanol were significantly decreased, which suggests that 1-1-tetradecanol has a potential inhibitory effect on the invasion and metastasis process of tumor cells, which may achieve this effect by interfering with the migration-related mechanisms of cells or affecting the biological characteristics of cells, providing important clues for subsequent in-depth exploration of its mechanism of action and application prospects in tumor treatment.

[0106] By detecting the changes in total ATP content after drug treatment, the ability of 1-1-tetradecanol to inhibit cell proliferation was indirectly explored. Since tumor cell proliferation requires a lot of energy to maintain, and ATP is the main source of cell energy; the specific experimental method is: collect drug-treated (5μM concentration, 2h) cells and blank control cell group (normal growth in DMEM high glucose medium environment), wash with PBS, and operate according to the instructions of Biyuntian ATP enhanced detection kit; combine Figures 9 and 10 After treating different tumor cells with 1-1-tetradecanol, the test results showed that the total ATP content in the drug-treated tumor cells changed significantly, that is, the ATP content decreased significantly. This suggests that the drug may reduce the energy supply of tumor cells by inhibiting the energy metabolism pathway of tumor cells, thereby changing the metabolic state of tumor cells to a certain extent. This is of great significance for understanding the mechanism of drug effects on tumor cell growth and survival, and also provides an experimental basis for subsequent research on the potential application value of the drug in tumor treatment.

[0107] To explore the effect of 1-1-tetradecanol on cell metabolism, the Seahorse experiment was used to compare the energy metabolism of each stage to further determine its effect on tumor cell proliferation. The specific experimental method was as follows: according to different treatment groups, the blank control group (cultured in DMEM high-glucose complete medium) and the 1-1-tetradecanol drug treatment group (concentration of 5μM, 30min) were divided into groups according to the instructions of the Seahorse exclusive kit; combined with Figure 11Part A is the U87MG SeahorseXF glycolysis stress test curve, and Part B is the U87MG SeahorseXF cell mitochondrial stress test curve.

[0108] Combine Figures 11 to 13 , studied the effects of U87MG under 1-1-tetradecanol treatment; combined with Figures 14 to 16 , studied the effects of U251 under 1-1-tetradecanol treatment. Figure 12 and Figure 14 In the figure, part A is the glycolytic rate, part B is the glycolytic capacity, and part C is the glycolytic reserve. n = 3. The results are expressed as mean ± standard deviation. * represents P < 0.05 compared with the control group, and ns represents P > 0.05 compared with the control group. U87C is the blank control group (Control group), and U87G is the 1-1-tetradecanol-treated group; Figure 13 and Figure 16 Part A is the basal respiration, part B is the ATP production, part C is the maximum respiration, and part D is the residual respiration. n = 3. The results are expressed as mean ± standard deviation. * represents P < 0.05 compared with the Control group. ns represents P > 0.05 compared with the Control group. U87C is the blank control group (Control group), and U87B is the 1-1-tetradecanol-treated group.

[0109] Many other changes and modifications can be made without departing from the spirit and scope of the present invention. It should be understood that the present invention is not limited to the specific embodiments, and the scope of the present invention is defined by the appended claims.

Claims

1. A method for extracting active substances from Herba Eupatorii, characterized by: The specific steps include: Step 100: Place the dried safflower grass into a crushing device for crushing; Step 200: Extract the crushed Herba Cistanches, saturated saline, and 2-octanol in an empty bottle, add a magnetic stirrer during the extraction, and seal the bottle in an SPME operating table. After the extraction, extract 1-tetradecanol extract from the Herba Cistanches.

2. The extraction method according to claim 1, wherein In step 200 , each extraction operation is preceded by a pre-equilibration at 70° C. for 10 minutes. The extraction operation includes solid phase microextraction, gas chromatography, and mass spectrometry analysis.

3. The extraction method according to claim 2, characterized in that In the extraction operation of step 200, the solid phase microextraction conditions are as follows: the aging extraction head is placed in the SPME operating table of the GERSTEL three-in-one autosampler for headspace adsorption for 20 minutes, and then the extraction head is inserted into the GC injection port for adsorption for 5 minutes, and the temperature program is started at the same time.

4. The extraction method according to claim 2, characterized in that In the extraction operation of step 200, the gas chromatography conditions are: injection port temperature 200°C, splitless injection, injection time 1 min, total flow rate 20 mL / min, purge flow rate 3 mL / min, and carrier gas He; Program temperature conditions: initial column temperature 30 °C, hold for 1 min, increase to 92 °C at a rate of 14 °C / min, hold for 2 min, increase to 200 °C at a rate of 5 °C / min, hold for 7 min, increase to 220 °C at a rate of 6 °C / min, hold for 8 min.

5. The extraction method according to claim 2, characterized in that In the extraction operation of step 200, the mass spectrometry conditions are as follows: ion source temperature 200°C; GC-MS interface temperature 200°C; solvent delay time 5 min; mass scan range (m / z) 35-500, full scan mode.

6. The extraction method according to claim 1, characterized in that The crushing device includes: A centrifugal grinder (6) and a cyclone separator (2) connected to the centrifugal grinder (6) through a connecting pipe (4), wherein a switching assembly (7) is provided at the bottom of the cyclone separator (2); The switching assembly (7) comprises a disc structure composed of a plurality of receiving bins (704) arranged along the circumference of a circle, the outer portion of the disc structure being connected to a discharge cover (5) for rotation, each receiving bin (704) being open to the outside and having an inclined screen (702) built therein, and the inclined screen (702) for screening materials being regularly replaced by the rotation of the disc structure; A coarse screen box (3) is provided between the equal-diameter cylinder (201) and the conical cylinder (203) of the cyclone separator (2), and a coarse screen (8) is installed in the coarse screen box (3) via a vibration assembly (9), and the vibration assembly (9) is linked downwardly to a rotatable disc structure via a push rod (901), and the disc structure is integrated with a square frame (701) that acts on the push rod (901), and the coarse screen (8) is prevented from vibrating and blocking by the rotation of the disc structure; The top of the discharge cover (5) is vertically connected to the conical cylinder (203) through a material guide opening (502); a return material interface (501) is provided on one side of the discharge cover (5); a fracture (503) is provided at the bottom thereof; the inclined screen (702) at the bottom end is cleaned and maintained through the fracture (503); large particles of the sedge grass in the receiving bin (704) are radially discharged through the return material interface (501); and small particles of the sedge grass pass through the inclined screen (702) and are axially discharged from the receiving bin (704).

7. The extraction method according to claim 6, characterized in that Square frames (701) are coaxially provided at both ends of the disc structure. A discharge port (705) connected to the receiving bin (704) is laterally provided on one side of the square frame (701), and the square frame (701) on the other side is connected to the first motor (703).

8. The extraction method according to claim 7, characterized in that It also includes a fine screening box (1) consisting of a box body (103), a disc structure is arranged inside the box body (103), and the end of the box body (103) is rotatably connected to the square frame (701) through a driven disc (102); A lateral opening (104) is provided on the side of the box body (103) at a position close to the discharge port (705), and a cleaning hole (101) communicating with the fracture (503) is provided on the bottom of the box body (103).

9. Application of active substances from Herba Eupatorii in anti-tumor medicine, which uses the extraction method of active substances from Herba Eupatorii according to claim 1, characterized in that: The active substance of euphorbia pulegosa is 1-tetradecanol.

10. Use of the active substance of Herba Eupatorii as an anti-tumor drug according to claim 9, characterized in that: Application of 1-tetradecanol in anti-glioma drugs.

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