Vehicle-mounted snake venom treatment device and method for treating snake venom using the same

Abstract

The application discloses a vehicle-mounted snake venom processing device and a method for processing snake venom by using the same. The vehicle-mounted snake venom processing device comprises a vehicle body, a workbench and a power supply device arranged in the vehicle body, and the workbench is provided with a separation device, a drying device, a detection device and a storage device. The drying device comprises a vehicle-mounted freeze dryer and an oil-free vacuum pump, and the crude snake venom separated by the separation device is subjected to freeze drying to obtain freeze-dried snake venom powder. The detection device comprises one or more of an enzyme label instrument, an ultraviolet spectrophotometer and a moisture tester for detecting biochemical indexes of the freeze-dried snake venom powder. The storage device is used for storing the freeze-dried snake venom powder which passes the detection. The vehicle-mounted snake venom processing device has a compact structure and is convenient to move and use, is suitable for collecting and processing snake venom in the scattered snake breeding of present snake breeders, significantly shortens the snake venom processing time, effectively retains active ingredients of the snake venom, improves the medicinal value of the snake venom, and is favorable for classified storage of the snake venom through on-site detection and analysis.

Description

Technical Field

[0001] This invention relates to the field of biomedical extraction technology, and in particular to a vehicle-mounted snake venom processing device and a method for processing snake venom using the same. Background Technology

[0002] Snake venom is a liquid secreted by venomous snakes from their venom glands. Proteins are the main toxic components of snake venom, accounting for approximately 90% to 95% of its dry weight. These proteins include two main categories: toxins and enzymes, classifying them as biotoxins. There are about twenty different types of enzymes and toxins in snake venom. In addition, it also contains some small peptides, amino acids, carbohydrates, lipids, nucleosides, biogenic amines, and metal ions. The composition of snake venom is extremely complex, and different snake venoms exhibit unique characteristics in terms of toxicity, pharmacology, and toxicological effects.

[0003] The medicinal value of snake venom is constantly being explored, and the quality requirements for snake venom are continuously increasing. With the deepening of research on snake venom, the quality requirements are gradually becoming more stringent. Currently, there is no unified standard for the quality evaluation and control of snake venom. Furthermore, traditionally, crude snake venom samples, after collection, generally need to be sent to a laboratory for analysis of biochemical parameters such as moisture content, protein content, phosphodiesterase, and L-amino acid oxidase. This process is difficult to process and preserve in a timely manner, leading to the mixing of crude snake venom samples of different purities, which affects the quality and stability of the crude venom and significantly increases the difficulty of further drug utilization. Therefore, there is a need to develop a mobile, vehicle-mounted snake venom processing device that can rapidly separate and test the quality of crude snake venom and efficiently preserve it, as well as a method for processing snake venom using this device. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to overcome the limitations of existing regions and the shortcomings of existing technologies, and to provide a mobile integrated vehicle-mounted snake venom treatment device that can separate and detect snake venom quality and is easy to use, as well as a method for treating snake venom using the vehicle-mounted snake venom treatment device.

[0005] One of the technical solutions adopted by this invention to solve its technical problem is:

[0006] A vehicle-mounted snake venom processing device includes a vehicle body, a workbench and a power supply unit installed inside the vehicle body. The workbench is equipped with a separation device, a drying device, a detection device and a storage device. The drying device includes a vehicle-mounted freeze dryer and an oil-free vacuum pump, which freeze-dries the crude snake venom obtained by the separation device to obtain freeze-dried snake venom powder. The detection device includes one or more of the following for detecting biochemical indicators of the freeze-dried snake venom powder: an enzyme-linked immunosorbent assay (ELISA) reader, an ultraviolet spectrophotometer, and a moisture analyzer. The storage device is used to store the qualified freeze-dried snake venom powder. The power supply unit is electrically connected to and supplies power to the separation device, the detection device, the drying device and the storage device.

[0007] This invention addresses the current situation where snake venom is collected in small quantities at a time, and most snake venom needs to be collected from breeding farms and transported back to processing farms for separation, drying, and laboratory testing. This results in snake venom not being processed in a timely manner, leading to damage to its active ingredients. The invention proposes a vehicle-mounted snake venom processing device, which facilitates timely processing of snake venom. The vehicle-mounted processing equipment also utilizes travel time between breeding farms to process snake venom samples, significantly shortening the processing time from collection to freeze-dried powder. Furthermore, the testing device can analyze the quality of different batches of snake venom, facilitating the classification and preservation of samples, especially removing substandard samples. This effectively avoids the blind collection of snake venom and improves the quality and stability of snake venom products. The vehicle-mounted structure of the snake venom processing device allows for rapid processing of snake venom from different breeding farms, improving equipment utilization. In addition, the vehicle-mounted snake venom processing device of this invention has a compact structure, high space utilization, and is easy to move and use.

[0008] Biochemical indicators of freeze-dried snake venom powder include water content, protein content, and enzyme activity. The enzyme activity determination method is based on the enzyme activity determination method described in Section III, Enzyme Activity Determination, of the article "Research and Utilization of Snake Venom I. Determination of Enzyme Activity in the Venom of Several Common Venomous Snakes in my country" (Chinese Journal of Biochemistry and Biophysics, June 1976, Vol. 8, No. 2), published by the Fourth Research Laboratory of Yunnan Institute of Zoology (Kunming).

[0009] The separation device is a vehicle-mounted centrifuge, the drying device is a freeze dryer, and the storage device is an ultra-low temperature freezer, preferably a -20℃ vehicle-mounted freezer.

[0010] The power supply unit includes a storage battery and a charging inverter. The storage battery and the original vehicle battery supply power to the separation device, drying device, detection device and storage device through the charging inverter to ensure the normal operation of the above equipment.

[0011] The vehicle-mounted snake venom treatment device also includes a snake venom collection device, which includes a refrigeration unit, a storage container, a vacuuming unit, and a venom extractor that penetrates the top of the refrigeration unit and extends into its interior for venom extraction. The bottom of the venom extractor is provided with an outer tube that communicates with the vacuuming unit and an embedded main tube that communicates with the storage container. The top of the venom extractor is provided with at least two layers of sterile flexible membrane. The storage container is placed inside the refrigeration unit and is sealed to the bottom of the venom extractor.

[0012] In one exemplary embodiment, the venom extractor has a filter element in its center. Preferably, a microporous membrane is placed on the filter element. The filter element inside the venom extractor can filter the snake venom fluid that has passed through the sterile flexible membrane, removing impurities. In particular, the filter element with the microporous membrane can not only filter impurities in the snake venom fluid, but also effectively maintain a negative pressure environment within the space enclosed by the storage container and the bottom of the venom extractor (especially when the sterile flexible membrane is torn by snake fangs after repeated use), promoting the rapid passage of the snake venom fluid through the venom extractor into the storage container.

[0013] In one exemplary embodiment, the poison extractor is any one of a sand core filtration funnel, a glass plate filtration funnel, or a filtration funnel with a cooling jacket, and the upper part of the poison extractor is a cylindrical body or a conical body.

[0014] The sterile flexible membrane has 2 to 20 layers, and the number of layers is proportional to the snake's tooth length and bite force. For example, when collecting venom from the five-step snake, a 14 to 20-layer sterile flexible membrane (using 3M sterile flexible membrane) is used; when collecting venom from the cobra, a 5 to 7-layer membrane is used. The multi-layered sterile flexible membrane can both stimulate the snake to actively spray venom and perform primary filtration of the venom, reducing the penetration of impurities.

[0015] In one exemplary embodiment, the sterile flexible membrane is secured to the top of the toxicant by a clamping component.

[0016] The clamping component is an elastic rubber band or a retaining ring with a tightening bolt.

[0017] The vacuum pumping device is an oil-free diaphragm vacuum pump.

[0018] The refrigeration device is a digitally displayed, temperature-controlled vehicle-mounted DC refrigerator, with a commonly used refrigeration temperature of -4°C. The storage container is preferably a ground-glass conical flask, but beakers, flasks, or test tubes can also be used. The refrigeration device includes a support plate to support the storage container, ensuring a secure connection between the storage container and the bottom of the extraction device. The top of the refrigeration device has a through-hole for the extraction device to pass through, and a sealing plug is installed at the through-hole to ensure a stable refrigeration environment.

[0019] In one exemplary embodiment, the snake venom collection device also includes a mobile cart, which is equipped with a frame for placing a vacuum device and a refrigeration device, so as to facilitate the rapid movement and use of the snake venom collection device.

[0020] The pore size of the microporous filter membrane is 20μm~100μm. For example, the mucus of the five-step snake is relatively thin, so a microporous filter membrane with a pore size of 20μm is used, while the mucus of the cobra is relatively thick, so a microporous filter membrane with a pore size of 100μm is used.

[0021] During use, the vacuum device maintains the vacuum level in the storage container and the toxicant at -0.03 to -0.05 MPa.

[0022] The vehicle also includes a fresh air system and an air conditioning system. The fresh air system can effectively filter microorganisms in the air and reduce the possibility of snake venom contamination. The air conditioning system consists of three parts: primary, medium and high efficiency. The air conditioning system maintains the internal temperature of the vehicle at 20-25℃ and the humidity at 50%-75%, which is conducive to the stability of experimental testing and the normal operation of experimental equipment.

[0023] Since snake venom loses its activity after being removed from the snake's body and remains at room temperature for several hours, and its toxicity disappears over time, this invention employs a mobile snake venom collection device. This device facilitates the collection of snake venom and allows for its refrigeration in a constant low-temperature environment. The collected venom is then rapidly processed using a vehicle-mounted processing device, which quickly centrifuges and freeze-dries the liquid snake venom into freeze-dried snake venom powder, extending its storage time and preserving its effective active ingredients.

[0024] A method for treating snake venom includes the following steps:

[0025] 1) Snake venom collection: Collect raw snake venom from snake farmers, or collect raw snake venom on-site using snake venom collection devices;

[0026] 2) Snake venom processing: The snake venom stock solution obtained in step 1) is separated using a separation device. The separated crude snake venom (supernatant after centrifugation) is then placed in a drying device for freeze drying to obtain freeze-dried snake venom powder. The biochemical indicators of the freeze-dried snake venom powder are then detected using a microplate reader and an ultraviolet spectrophotometer. Based on the detected biochemical indicators, the freeze-dried snake venom powder is classified and placed in a storage device for refrigeration.

[0027] In step 1), the specific operation of collecting raw snake venom solution on-site using a snake venom collection device is as follows:

[0028] a. Prepare the snake venom collection device: Determine the number of sterile flexible membrane layers based on the tooth length and bite force of the snake species from which the venom will be collected; then assemble the snake venom collection device—place the storage container inside the refrigeration unit, then seal the venom extractor to the top of the storage container, then connect the outer tube of the venom extractor to the vacuum device via a flexible tube, lay a microporous filter membrane inside the venom extractor, then lay multiple layers of sterile flexible membrane on the top of the venom extractor and secure it with clamping components; start the refrigeration unit and adjust the temperature of the refrigeration unit to the specified refrigeration temperature;

[0029] b. Venom collection: Activate the vacuum device, then bring the snake's head into contact with the sterile flexible membrane at the top of the venom extractor to induce the snake to actively open its mouth and bite the sterile flexible membrane. The snake's teeth will insert into the membrane and automatically spray out venom. Under the negative pressure suction effect created by the vacuum device, the venom will quickly pass through the microporous filter membrane and flow into the storage container along the embedded main pipe of the venom extractor.

[0030] c. Return the snake to its enclosure after collecting venom.

[0031] Preferably, in step b, before starting the vacuum device, an enzyme protectant for stabilizing snake venom activity is added to the storage container. More preferably, a thrombin-like protectant is added. The thrombin-like protectant is selected from one or more of the following: amino acids such as glycine, dextran, trehalose, human serum albumin, and hydrolyzed gelatin. The enzyme protectant is stored at 2-8°C and added to the storage container before use. By adding the enzyme protectant to the storage container, the enzyme activity in the snake venom is maintained. Compared to storage without an enzyme protectant, the snake venom enzyme activity is nearly doubled in this method of protection.

[0032] In step 2), the freeze-drying time is 4~8 hours, and the freeze-dried snake venom powder has a water content ≤5%, L-amino acid oxidase activity ≥15~25 U / mg, alkaline phosphatase activity ≥10~25 U / mg, phosphodiesterase activity ≥15~30 U / mg, and ribonuclease activity ≥20~30 U / mg.

[0033] The beneficial effects of the vehicle-mounted snake venom treatment device of the present invention:

[0034] The vehicle-mounted snake venom processing device has a compact structure, high space utilization, and is easy to move and use, making it suitable for the collection and processing of snake venom by snake farmers in their dispersed operations. Compared with conventional snake venom collection and processing methods, it significantly shortens the processing time, effectively preserves the active ingredients of the snake venom, and improves its medicinal value. Furthermore, on-site testing and analysis facilitate the classification and preservation of snake venom, making it easier to classify, preserve, and apply snake venom based on its type, purity, or enzyme activity.

[0035] This vehicle-mounted snake venom processing device utilizes a mobile snake venom collection unit and a vacuum device to create a slight negative pressure inside the storage container and sampler. This accelerates the flow of snake venom automatically sprayed from the snake into the storage container, which is beneficial for the retention of active ingredients in the snake venom and reduces the amount of snake venom residue on the inner wall of the sampler, thereby improving the efficiency and yield of snake venom collection.

[0036] Secondly, the venom collection device has a microporous filter membrane on the filter element inside the venom extractor to filter the snake venom again, reducing the impurity content of the snake venom entering the storage container and improving the purity of the snake venom.

[0037] In addition, the snake venom collection device is easy to use and has a simple structure. It uses a digital display temperature-controlled vehicle-mounted DC refrigerator to provide a constant environment for storing snake venom, preventing the snake venom from losing its activity and improving the retention rate of enzyme activity.

[0038] The beneficial effects of this invention's snake venom treatment method using a vehicle-mounted snake venom treatment device are as follows:

[0039] This method is simple and efficient, and can quickly collect, separate, test and preserve snake venom, effectively retaining the active ingredients of snake venom and improving its medicinal value. Attached Figure Description

[0040] Figure 1 —This is a schematic diagram of the structure of a vehicle-mounted snake venom treatment device in Example 1;

[0041] Figure 2 —A top view of the interior of the vehicle body in Embodiment 1;

[0042] Figure 3 —This is a schematic diagram of the snake venom collection device of a vehicle-mounted snake venom treatment device in Example 1;

[0043] Figure 4 -for Figure 3 A schematic diagram of the structure of the poison extraction device;

[0044] Figure 5 —A top view of the interior of the vehicle body of a vehicle-mounted snake venom treatment device in Embodiment 2;

[0045] Figure 6 —A top view of the interior of the vehicle body of a vehicle-mounted snake venom treatment device in Embodiment 3;

[0046] Figure 7 —This is a schematic diagram of the snake venom collection device of a vehicle-mounted snake venom treatment device in Example 3;

[0047] Figure 8 -for Figure 7 A schematic diagram of the poison extraction device.

[0048] In the diagram: 1. Vehicle body; 2. Storage device; 3. Detection device; 31. Spectrophotometer; 32. Microplate reader; 33. Moisture analyzer; 34. pH meter; 35. Analytical balance; 4. Workbench; 5. Drying device; 51. Vehicle-mounted freeze dryer; 52. Oil-free vacuum pump; 6. Separation device; 7. Power supply device; 8. Snake venom collection device; 81. Sterile flexible membrane; 82. Clamping component; 83. Venom extractor; 831. Filter element; 832. Outer tube; 833. Embedded main tube; 834. Ground joint connector; 835. Cooling jacket; 84. Storage container; 85. Refrigeration device; 851. Support plate; 852. Sealing plug; 86. Vacuum pump; 87. Mobile trolley. Detailed Implementation

[0049] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0050] Example 1

[0051] Reference Figures 1-4 The vehicle-mounted snake venom processing device of this embodiment includes a vehicle body 1 and a workbench 4, a power supply device 7, and a snake venom collection device 8 installed inside the vehicle body 1. The vehicle body 1 includes a base, a driver's cab, and a cargo compartment. The snake venom collection device 8 is placed at the exit of the cargo compartment (the cargo compartment exit is located at the rear of the cargo compartment). The workbench 4 is installed inside the cargo compartment. The workbench 4 is equipped with a separation device 6, a drying device 5, a detection device 3, and a storage device 2. The drying device 5 includes a vehicle-mounted freeze dryer 51 and an oil-free vacuum pump 52, which freeze-dries the crude snake venom separated by the separation device 6 to obtain freeze-dried snake venom powder. The detection device 3 includes an enzyme-linked immunosorbent assay (ELISA) reader 32, an ultraviolet spectrophotometer, and a moisture analyzer 33 for detecting the biochemical indicators of the freeze-dried snake venom powder. The storage device 2 is used to store the qualified freeze-dried snake venom powder. The power supply device 7 is electrically connected to and supplies power to the separation device 6, the detection device 3, the drying device 5, and the storage device 2.

[0052] The biochemical indicators of freeze-dried snake venom powder include its moisture content, protein content, and enzyme activity. The enzyme activity determination method was based on the enzyme activity determination method described in Section III, Enzyme Activity Determination, of the article "Research and Utilization of Snake Venom I. Determination of Enzyme Activity in the Venom of Several Common Venomous Snakes in my country" (Acta Biochimica & Biophysics, June 1976, Vol. 8, No. 2), published by the Fourth Research Laboratory of the Yunnan Provincial Institute of Zoology (Kunming). Enzyme activity was determined using a UV spectrophotometer, mainly targeting L-amino acid oxidase, alkaline phosphatase, phosphodiesterase, and ribonuclease. The specific procedures are as follows:

[0053] L-amino acid oxidase activity was determined using the Nessler reagent method for free ammonia. The reaction system consisted of: 0.2 mL of 1 mg / mL snake venom solution (prepared in 0.1 M Tris-HCl buffer, pH 7.6), 1 mL of 0.1 M L-leucine (prepared in the same buffer), for a total reaction volume of 1.2 mL. After incubation at 37°C for 2 hours, 1 mL of 27% trichloroacetic acid was added to stop the reaction. After standing for 20 minutes, the mixture was filtered. 1 mL of the filtrate was added to 1 mL of distilled water and 3 mL of Nessler reagent solution, and the ammonia production was determined colorimetrically at 430 nm.

[0054] Alkaline phosphodiesterase and phosphodiesterase activities were determined using the Bessey method. The reaction system consisted of: 0.2 mL of 1 mg / mL snake venom solution (prepared in 0.05 M glycine buffer containing 1.2 × 10⁻² M MgCl₂, pH 8.8), 1.0 mL of 1.2 × 10⁻² M disodium p-nitrophenyl phosphate or di-p-nitrophenyl phosphate (prepared in the above buffer, pH 8.8), for a total reaction volume of 1.2 mL. After incubation at 37 °C for 5–40 minutes, 10 mL of 0.02 N NaOH was added to stop the reaction. The formation of p-nitrophenol was measured colorimetrically at 420 nm.

[0055] Ribonuclease activity was determined according to the McDonald method. The reaction system consisted of: 0.2 mL of 5–8 mg / mL snake venom solution (prepared in 0.1 M Tris-HCl buffer, pH 7.6), 1.0 mL of 7 mg / mL yeast solution (prepared in the above buffer), for a total reaction volume of 1.2 mL. After incubation at 30 °C for 30 minutes, 2 mL of MacFadyen's reagent was added to stop the reaction. After standing for 30 minutes, the mixture was filtered. 1 mL of the filtrate was added to 1 mL of distilled water and 6 mL of 0.2% 3,5-dihydroxytoluene solution. The mixture was heated in a boiling water bath for 20 minutes for color development. The oligonucleotides were measured at 680 nm.

[0056] The enzyme activity of the freeze-dried snake venom powder was determined using a 72-type spectrophotometer (31) manufactured by Shanghai Analytical Instrument Factory; its protein content was determined using an enzyme-linked immunosorbent assay (ELISA) reader (32); and its moisture content was rapidly determined using a moisture analyzer (33). The composition and activity of the freeze-dried snake venom powder, separated and freeze-dried using a vehicle-mounted snake venom processing device, were tested to remove substandard raw materials and facilitate the classification and storage of the venom according to its source, type, and enzyme activity.

[0057] The separation device 6 is a vehicle-mounted centrifuge, optionally the Kaida TDL5M vehicle-mounted centrifuge. The drying device 5 is a freeze dryer (such as the Linhai Tanshi Vacuum Equipment Co., Ltd. oil-free vacuum pump freeze dryer SPH-20). The storage device 2 is an ultra-low temperature freezer, preferably a -20℃ vehicle-mounted freezer, to reduce the impact of storage and regulation on the active ingredients during snake venom transportation.

[0058] The power supply device 7 includes a storage battery and a charging inverter. The storage battery and the original vehicle battery supply power to the separation device 6, the drying device 5, the detection device 3 and the storage device 2 through the charging inverter to ensure the normal use of the above equipment.

[0059] The vehicle body 1 also includes a fresh air system and an air conditioning system. The fresh air system facilitates efficient filtration of microorganisms in the air, reducing the possibility of snake venom contamination. The air conditioning system consists of three parts: primary, medium and high efficiency. The air conditioning system maintains the internal temperature of the vehicle body at 20-25℃ and the humidity at 50%-75%, which facilitates the stability of experimental testing and the normal operation of experimental equipment.

[0060] The snake venom collection device 8 includes a refrigeration device 85, a storage container 84, a vacuum device 86, and a venom extractor 83 that penetrates the top of the refrigeration device 85 and extends into it for venom extraction. The bottom of the venom extractor 83 is provided with an outer tube 832 that communicates with the vacuum device 86 and an embedded main tube 833 that communicates with the storage container 84. The top of the venom extractor 83 is provided with at least two layers of sterile flexible membrane 81. The storage container 84 is placed inside the refrigeration device 85 and is sealed to the bottom of the venom extractor 83.

[0061] The venom extractor 83 has a filter element 831 in the middle, and a microporous filter membrane is placed on the filter element 831. The pore size of the microporous filter membrane is 20μm~100μm. For example, the mucus of the five-step snake is relatively thin, so a microporous filter membrane with a pore size of 20μm is used, while the mucus of the cobra is relatively thick, so a microporous filter membrane with a pore size of 100μm is used.

[0062] The filter element 831 inside the venom extractor 83 can filter the snake venom fluid that has passed through the sterile flexible membrane 81, removing impurities. In particular, the filter element 831 with a microporous membrane can not only filter impurities in the snake venom fluid, but also effectively maintain the negative pressure environment in the space enclosed by the storage container 84 and the bottom of the venom extractor 83 compared to a venom extractor 83 without a filter element 831 or without a microporous membrane filter element 831 (especially when the sterile flexible membrane 81 is torn by snake fangs after repeated use), promoting the rapid passage of the snake venom fluid through the venom extractor 83 into the storage container 84.

[0063] The poison extractor 83 is a sand core filter funnel, and the upper part of the poison extractor 83 is a cylindrical body.

[0064] The sterile flexible membrane 81 has 2 to 20 layers, and the number of layers is proportional to the snake's tooth length and bite force. For example, when collecting venom from the five-step snake, 14 to 20 layers of sterile flexible membrane 81 (using 3M sterile flexible membrane 81) are used; when collecting venom from the cobra, 5 to 7 layers are used. The multi-layered sterile flexible membrane 81 can both stimulate the snake to actively spray venom and perform primary filtration of the venom, reducing the penetration of impurities.

[0065] The sterile flexible membrane 81 is fixed to the top of the poison extractor 83 by the clamping component 82.

[0066] The clamping component 82 is a retaining ring with a tightening bolt.

[0067] The vacuum pumping device 86 is an oil-free diaphragm vacuum pump.

[0068] The refrigeration device 85 is a digital display temperature-controlled vehicle-mounted DC refrigerator, with a commonly used refrigeration temperature of -4℃. The storage container 84 is preferably a ground glass conical flask, but beakers, flasks, or test tubes can also be used. The refrigeration device 85 has a support plate 851 for supporting the storage container 84, and it is well connected to the ground glass joint 834 at the bottom of the storage container 84 and the poison extractor 83. Its sealing performance is good, ensuring the sealing conditions required for the vacuuming device 86 to create a vacuum. The top of the refrigeration device 85 has a through hole for the poison extractor 83 to pass through, and a sealing plug 852 is provided at the through hole to ensure that the refrigeration device 85 provides a stable refrigeration environment.

[0069] The snake venom collection device 8 also includes a mobile trolley 87, which is equipped with a frame for placing a vacuum device 86 and a refrigeration device 85, so as to facilitate the rapid movement and use of the snake venom collection device 8.

[0070] During use, the vacuum device 86 maintains the vacuum level in the storage container 84 and the poison extractor 83 at -0.03 to -0.05 MPa.

[0071] A method for treating snake venom includes the following steps:

[0072] 1) Snake venom collection: Remove the snake venom collection device 8 from vehicle 1 and transport it to the snake breeding base on-site using a mobile cart 87 to collect raw snake venom. The specific operation is as follows:

[0073] a. Prepare the snake venom collection device 8: Determine the number of layers of the sterile flexible membrane 81 based on the tooth length and bite force of the snake species from which the venom is to be collected; then assemble the snake venom collection device 8—place the storage container 84 into the refrigeration device 85, then seal the venom extractor 83 to the top of the storage container 84, then connect the outer tube 832 of the venom extractor 83 to the vacuum device 86 through a flexible tube, lay a microporous filter membrane inside the venom extractor 83, then lay multiple layers of sterile flexible membrane 81 on the top of the venom extractor 83 and fix it with the clamping component 82; start the refrigeration device 85 and adjust the temperature of the refrigeration device 85 to the specified refrigeration temperature;

[0074] b. Venom Collection: First, an enzyme protectant for stabilizing snake venom activity is added to the storage container 84. The enzyme protectant is selected from one or more of the following: amino acids such as glycine, dextran, trehalose, human serum albumin, and hydrolyzed gelatin. For example, 1% glycine is added to the storage container 84 as an enzyme protectant. The enzyme protectant is stored at 2-8℃ and added to the storage container 84 before use. Then, the vacuum device 86 is activated, and the snake's head is brought into contact with the sterile flexible membrane 81 at the top of the venom extractor 83 to induce the snake to actively open its mouth and bite the sterile flexible membrane 81. The snake's teeth are inserted into the membrane and the venom is automatically sprayed out. Under the negative pressure suction effect formed by the vacuum device 86, the venom is quickly filtered through the microporous filter membrane and flows into the storage container 84 along the embedded main pipe 833 of the venom extractor 83.

[0075] c. Return the snake to its enclosure after collecting venom.

[0076] 2) Snake venom processing: The snake venom stock solution obtained in step 1) is separated using the separation device 6. The separated crude snake venom (supernatant after centrifugation) is then placed in the drying device 5 for freeze-drying for 8 hours to obtain snake venom freeze-dried powder. The biochemical indicators of the snake venom freeze-dried powder are then detected using a microplate reader 32 and an ultraviolet spectrophotometer 3. Based on the detected biochemical indicators, the snake venom freeze-dried powder is classified and placed in the storage device 2 for refrigeration.

[0077] According to the test results, the freeze-dried snake venom powder obtained by the snake venom treatment method in this embodiment has a water content of 3%, an L-amino acid oxidase activity of 25 U / mg, an alkaline phosphatase activity of 20 U / mg, a phosphodiesterase activity of 28 U / mg, and a ribonuclease activity of 30 U / mg.

[0078] Example 2

[0079] Reference Figure 5The vehicle-mounted snake venom processing device of this embodiment includes a vehicle body 1 and a workbench 4, a power supply device 7, and a snake venom collection device 8 installed inside the vehicle body 1. The vehicle body 1 includes a base, a driver's cab, and a cargo compartment. The workbench 4 is located in the cargo compartment near the exit. The workbench 4 is equipped with a separation device 6, a drying device 5, a detection device 3, and a storage device 2. The drying device 5 includes a vehicle-mounted freeze dryer 51 and an oil-free vacuum pump 52, which freeze-dries the crude snake venom obtained by the separation device 6 to obtain freeze-dried snake venom powder. The detection device 3 includes an enzyme-linked immunosorbent assay (ELISA) reader 32, an ultraviolet spectrophotometer 31, and an electronic scale for detecting the biochemical indicators of the freeze-dried snake venom powder. The storage device 2 is used to store the qualified freeze-dried snake venom powder and is located at the end of the cargo compartment away from the exit. The power supply device 7 is electrically connected to and supplies power to the separation device 6, the detection device 3, the drying device 5, and the storage device 2.

[0080] The detection device 3 in this embodiment can also be configured with conventional detection equipment as needed, such as an analytical balance 35, a pH meter 34, etc.

[0081] The separation device 6 is a vehicle-mounted centrifuge, the drying device 5 is a freeze dryer, and the storage device 2 is an ultra-low temperature freezer, preferably a -20℃ vehicle-mounted freezer.

[0082] The power supply device 7 includes a storage battery and a charging inverter. The storage battery and the original vehicle battery supply power to the separation device 6, the drying device 5, the detection device 3 and the storage device 2 through the charging inverter to ensure the normal use of the above equipment.

[0083] A method for treating snake venom includes the following steps:

[0084] 1) Snake venom collection: Collect raw snake venom (cobra venom) from snake farmers, add 1% glycine as an enzyme protectant to the raw snake venom, and refrigerate the raw snake venom at 4°C.

[0085] 2) Snake venom processing: The snake venom stock solution obtained in step 1) is separated using the separation device 6. The separated crude snake venom (supernatant after centrifugation) is then placed in the drying device 5 for freeze-drying for 6 hours to obtain snake venom freeze-dried powder. The biochemical indicators of the snake venom freeze-dried powder are then detected using a microplate reader 32 and an ultraviolet spectrophotometer 3. Based on the detected biochemical indicators, the snake venom freeze-dried powder is classified and placed in the storage device 2 for refrigeration.

[0086] According to the test results, the freeze-dried snake venom powder obtained by the snake venom treatment method in this embodiment has a water content of 4%, an L-amino acid oxidase activity of 18 U / mg, an alkaline phosphatase activity of 20 U / mg, a phosphodiesterase activity of 22 U / mg, and a ribonuclease activity of 21 U / mg.

[0087] Example 3

[0088] Reference Figures 6-8 The vehicle-mounted snake venom treatment device of this embodiment includes a vehicle body 1 and a workbench 4, a power supply device 7, and a snake venom collection device 8 installed inside the vehicle body 1. The vehicle body 1 includes a base, a driver's cab, and a cargo compartment. The snake venom collection device 8 is fixedly installed at the exit of the cargo compartment (the cargo compartment exit is located at the rear of the cargo compartment). The workbench 4 is installed inside the cargo compartment. On the workbench 4, from the outside to the inside of the cargo compartment exit, a separation device 6, a drying device 5, a detection device 3, and a storage device 2 are arranged sequentially to facilitate the rapid separation, drying, and detection of the collected snake venom. The process includes testing and storage; the drying device 5 includes a vehicle-mounted freeze dryer 51 and an oil-free vacuum pump 52, which freeze-dries the crude snake venom obtained by the separation device 6 to obtain freeze-dried snake venom powder; the detection device 3 includes an enzyme-linked immunosorbent assay (ELISA) reader 32, an ultraviolet spectrophotometer, a moisture analyzer 33, and an electronic balance for detecting the biochemical indicators of the freeze-dried snake venom powder; the storage device 2 is used to store the qualified freeze-dried snake venom powder; the power supply device 7 is electrically connected to and supplies power to the separation device 6, the detection device 3, the drying device 5, and the storage device 2.

[0089] The separation device 6 and power supply device 7 are the same as in Embodiment 1.

[0090] The snake venom collection device 8 includes a refrigeration device 85, a storage container 84, a vacuum device 86, and a venom extractor 83 that penetrates the top of the refrigeration device 85 and extends into it for venom extraction. The bottom of the venom extractor 83 is provided with an outer tube 832 that communicates with the vacuum device 86 and an embedded main tube 833 that communicates with the storage container 84. The top of the venom extractor 83 is provided with at least two layers of sterile flexible membrane 81. The storage container 84 is placed inside the refrigeration device 85 and is sealed to the bottom of the venom extractor 83.

[0091] The venom extractor 83 is a glass plate suction funnel with a cooling jacket 835, and the upper part of the venom extractor 83 is a cylindrical body. The cooling jacket 835 of the venom extractor 83 is connected to a cooling water circulator, and the temperature of the cooling water inside is preferably 2~8℃. This is more conducive to the retention of the effective active ingredients in the snake venom solution flowing into the venom extractor 83, especially in summer when the outside temperature is high. The glass plate suction funnel with the cooling jacket 835, as the venom extractor 83, is rapidly cooled by the cooling water pumped by the cooling water circulator, so that the venom extractor 83 quickly enters a low-temperature environment after leaving the snake venom gland, thus slowing down the inactivation rate of the effective ingredients in the snake venom during the collection process. The cooling water circulator can be a CW3000 laser chiller. The venom extractor 83 has a filter element 831 in the middle, and a microporous filter membrane is placed on the filter element 831. The pore size of the microporous filter membrane is 20μm~100μm. For example, the mucus of the five-step snake is relatively thin, so a microporous filter membrane with a pore size of 20μm is used, while the mucus of the cobra is relatively thick, so a microporous filter membrane with a pore size of 100μm is used.

[0092] The sterile flexible membrane 81 has 2 to 20 layers, and the number of layers is proportional to the length of the snake's teeth and the bite force. In this embodiment, the snake venom collection device 8 is used for collecting venom from the five-step snake. Since the teeth of the five-step snake are relatively long, basically 8-16 mm, the number of layers of the sterile flexible membrane 811 is 14 to 20, and the number of layers is proportional to the length of the snake's teeth and the bite force. This makes the thickness of the multi-layer sterile flexible membrane 811 slightly smaller than the length of the five-step snake's teeth by 1 to 2 mm. Thus, the multi-layer sterile flexible membrane 811 can both stimulate the snake to actively spray venom and perform primary filtration of the snake venom, reducing the penetration of impurities.

[0093] When the snake venom collection device 8 of this embodiment is used for collecting snake venom from cobras, since cobras have relatively short teeth, generally between 6 and 9 millimeters, the sterile flexible membrane 811 has 5 to 7 layers, which are proportional to the length of the snake's teeth and its biting force. This makes the thickness of the multi-layered sterile flexible membrane 811 slightly smaller than the length of the cobra's teeth by 0.5 to 1 millimeter. Thus, the multi-layered sterile flexible membrane 811 can both stimulate the snake to actively spray venom and perform primary filtration of the snake venom, reducing the penetration of impurities.

[0094] When the snake venom collection device 8 of this embodiment is used to collect snake venom from the banded krait, since the banded krait has the shortest teeth among snakes, basically 2-4 mm, and the sterile flexible membrane 811 has 2-4 layers, which are proportional to the length of the snake's teeth and the biting force, so that the thickness of the multi-layer sterile flexible membrane 811 is slightly smaller than the length of the cobra's teeth by 0.5-1 mm. Thus, the multi-layer sterile flexible membrane 811 can both stimulate the snake to actively spray venom and perform primary filtration of the snake venom, reducing the penetration of impurities.

[0095] The other components of the snake venom collection device 8 are the same as those described in Embodiment 1, and will not be repeated here.

[0096] A method for treating snake venom includes the following steps:

[0097] 1) Snake venom collection: Drive the snake venom processing device to the snake breeding area and use the snake venom collection device 8 to collect raw snake venom solution on site. The specific operation is as follows:

[0098] a. Prepare the snake venom collection device 8: Determine the number of layers of the sterile flexible membrane 81 based on the tooth length and bite force of the snake species from which the venom is to be collected; then assemble the snake venom collection device 8—place the storage container 84 into the refrigeration device 85, then seal the venom extractor 83 to the top of the storage container 84, then connect the outer tube 832 of the venom extractor 83 to the vacuum device 86 through a flexible tube, lay a microporous filter membrane inside the venom extractor 83, then lay multiple layers of sterile flexible membrane 81 on the top of the venom extractor 83 and fix it with the clamping component 82; start the refrigeration device 85 and adjust the temperature of the refrigeration device 85 to the specified refrigeration temperature;

[0099] b. Venom collection: First, add 1% glycine to the storage container 84 as an enzyme protectant to stabilize the activity of snake venom. The enzyme protectant is stored at 2~8℃ and is added to the storage container 84 when in use. Then, start the vacuum device 86 and manually bring the snake's head into contact with the sterile flexible membrane 81 at the top of the venom extractor 83 to induce the snake to actively open its mouth and bite the sterile flexible membrane 81. The snake's teeth are inserted into the membrane and the venom is automatically sprayed out. Under the negative pressure suction effect formed by the vacuum device 86, the venom is quickly filtered through the microporous filter membrane and flows into the storage container 84 along the embedded main pipe 833 of the venom extractor 83.

[0100] c. Return the snake to its enclosure after collecting venom.

[0101] 2) Snake venom processing: The snake venom stock solution obtained in step 1) is separated using the separation device 6. The separated crude snake venom (supernatant after centrifugation) is then placed in the drying device 5 for freeze-drying for 4 hours to obtain snake venom freeze-dried powder. The biochemical indicators of the snake venom freeze-dried powder are then detected using a microplate reader 32 and an ultraviolet spectrophotometer 3. Based on the detected biochemical indicators, the snake venom freeze-dried powder is classified and placed in the storage device 2 for refrigeration.

[0102] According to the test results, the freeze-dried snake venom powder obtained by the snake venom treatment method in this embodiment has a water content of 4.2%, an L-amino acid oxidase activity of 22 U / mg, an alkaline phosphatase activity of ≥ 25 U / mg, an alkaline phosphatase activity of ≥ 25 U / mg, and an alkaline phosphatase activity of ≥ 28 U / mg.

[0103] This invention discloses a vehicle-mounted snake venom processing device. The detection device 3 can be supplemented with appropriate equipment based on the physicochemical parameters of the freeze-dried snake venom powder to be tested. For example, when it is necessary to analyze the components in the freeze-dried snake venom powder, a liquid chromatography analyzer can be added. The number of layers of the sterile flexible membrane 81 on the venom extractor 83 can be changed according to the snake venom collected from different snakes, and is not limited to the number of layers exemplified in the above embodiments. These changes in technical features can be understood and implemented by those skilled in the art through the textual description, and therefore will not be further illustrated with accompanying drawings. In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

Claims

1. A vehicle-mounted snake venom treatment device, characterized in that, The device includes a vehicle body (1) and a workbench (4) and a power supply device (7) installed inside the vehicle body (1). The workbench (4) is equipped with a separation device (6), a drying device (5), a detection device (3), and a storage device (2). The drying device (5) includes a vehicle-mounted freeze dryer (51) and an oil-free vacuum pump (52) to freeze-dry the crude snake venom obtained by the separation device (6) to obtain freeze-dried snake venom powder. The detection device (3) includes one or more of an enzyme-linked immunosorbent assay (ELISA) reader (32), an ultraviolet spectrophotometer (31), and a moisture analyzer (33) for detecting the biochemical indicators of the freeze-dried snake venom powder. The storage device (2) is used to store the qualified freeze-dried snake venom powder. The power supply device (7) is electrically connected to and supplies power to the separation device (6), the detection device (3), the drying device (5), and the storage device (2). The vehicle-mounted snake venom treatment device also includes a snake venom collection device (8), which includes a refrigeration device (85), a storage container (84), a vacuum device (86), and a venom extractor (83) that penetrates the top of the refrigeration device (85) and extends into it for venom extraction. The venom extractor (83) has an outer tube (832) at the bottom that communicates with the vacuum device (86) and an embedded main tube (833) that communicates with the storage container (84). The top of the venom extractor (83) is provided with at least two layers of sterile flexible membrane (81). The storage container (84) is placed inside the refrigeration device (85) and is sealed to the bottom of the venom extractor (83). The venom extractor (83) is a glass plate suction funnel with a cooling jacket (835). The upper part of the venom extractor (83) is a cylindrical body. The cooling jacket (835) of the venom extractor (83) is connected to a cooling water circulator. The biochemical indicators of the freeze-dried snake venom powder were detected using the detection device (3). Based on the detected biochemical indicators, the freeze-dried snake venom powder was classified and placed in the storage device (2) for refrigeration.

2. The vehicle-mounted snake venom treatment device as described in claim 1, characterized in that, The separation device (6) is a vehicle-mounted centrifuge, the drying device (5) is a freeze dryer, and the storage device (2) is an ultra-low temperature freezer.

3. The vehicle-mounted snake venom treatment device as described in claim 1, characterized in that, The power supply device (7) includes a storage battery and a charging inverter. The storage battery and the original vehicle battery supply power to the separation device (6), the drying device (5), the detection device (3) and the storage device (2) through the charging inverter.

4. The vehicle-mounted snake venom treatment device as described in claim 1, characterized in that, The sterile flexible membrane (81) has 2 to 20 layers, and the number of layers is proportional to the length of the snake's teeth and the biting force.

5. The vehicle-mounted snake venom treatment device as described in claim 1, characterized in that, The poison extractor (83) has a filter core (831) in the middle, and a microporous filter membrane is placed on the filter core (831).

6. The vehicle-mounted snake venom treatment device as described in claim 1, characterized in that, The sterile flexible membrane (81) is fixed to the top of the poison extractor (83) by a clamping component (82).

7. The vehicle-mounted snake venom treatment device as described in any one of claims 1 to 6, characterized in that, The snake venom collection device (8) also includes a mobile trolley (87), which has a frame for placing a vacuum device (86) and a refrigeration device (85).

8. A method for treating snake venom using a vehicle-mounted snake venom treatment device, characterized in that, The vehicle-mounted snake venom treatment device as described in any one of claims 1 to 7, wherein the method comprises the following steps: 1) Snake venom collection: Collect raw snake venom from snake farmers, or collect raw snake venom on-site using a snake venom collection device (8); 2) Snake venom treatment: The snake venom raw liquid obtained in step 1) is separated by a separation device (6), and the separated crude snake venom is placed in a drying device (5) for freeze drying to obtain snake venom freeze-dried powder. The biochemical indicators of the snake venom freeze-dried powder are then detected by a detection device (3). Based on the detected biochemical indicators, the snake venom freeze-dried powder is classified and placed in a storage device (2) for refrigeration.

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