High-efficiency rapid-coagulation blood coagulant containing thrombin

Abstract

The invention relates to the field of hematology examination, and discloses a thrombin-containing high-efficiency rapid-coagulation blood coagulant which is prepared by the following steps: measuring 1000ml of an absolute ethyl alcohol solution, putting the absolute ethyl alcohol solution into a beaker, and putting the beaker on a magnetic stirrer; weighing 30-80g of silicon dioxide powder, adding the silicon dioxide powder into the absolute ethyl alcohol solution, starting magnetic stirring, weighing 10-60ml of thrombin, adding the thrombin into the absolute ethyl alcohol solution, continuously stirring, finally adding 1-20ml of an inner wall coating agent, and stirring for 10 minutes. By adopting the high-efficiency rapid-coagulation blood coagulant containing thrombin, the technical effect of remarkably increasing the blood coagulation speed is achieved. Compared with the technical scheme that an inorganic coagulant is used in the prior art, the blood can be rapidly coagulated within 3-5 minutes through direct participation of thrombin, so that delay caused by too long blood coagulation time in clinical and laboratory detection is greatly reduced, and the overall efficiency of blood sample treatment is improved.

Description

Technical Field

[0001] The invention relates to the field of hematological testing, in particular to a high-efficiency fast-clotting blood coagulant containing thrombin. Background Art

[0002] In early medical tests, obtaining accurate blood test results requires that blood samples coagulate as quickly as possible and separate components such as serum or plasma for analysis. However, the body's natural coagulation process is relatively slow and may take several minutes or even longer to complete coagulation, which is inefficient for clinical practice and scientific research that requires rapid test results. Therefore, in order to improve the speed and efficiency of blood sample processing, the demand for substances that can accelerate blood coagulation is increasing, which has promoted the research and development and application of blood coagulants.

[0003] After in-depth research on the coagulation mechanism of the human body, we know that the coagulation process is a series of complex chemical reactions involving the activation and interaction of multiple coagulation factors. Based on this, our company began to explore chemical substances that can activate or promote these coagulation reactions, and found that the substance thrombin plays a key role in the coagulation process. Thrombin can convert fibrinogen in the blood into fibrin, which interweaves to form a network structure, causing the blood to coagulate and form blood clots, thereby achieving the purpose of hemostasis.

[0004] Thrombin is a natural component of the body's coagulation system. It consists of two peptide chains connected by disulfide bonds. Thrombin exists in the body in the form of prothrombin. Under certain conditions, thrombin is activated and converted into an active protease.

[0005] Currently, the blood coagulants on the market are mainly divided into two types, those containing thrombin and those not containing thrombin.

[0006] The coagulant that does not contain thrombin is inorganic, and its main components are silicon dioxide powder, diatomaceous earth, silica powder, etc. Its function is to activate the body's internal and external coagulation systems.

[0007] The disadvantage of inorganic coagulants is that the coagulation speed is relatively slow: compared with biological coagulants, inorganic coagulants do not activate the coagulation system quickly enough, and it takes a relatively long time to complete the coagulation process. Fibrin is prone to secondary precipitation: after using inorganic coagulants, fibrin may precipitate from the serum again during subsequent processing or storage, which will affect the quality of the blood sample and subsequent detection and analysis.

[0008] Weak fibrin compression function: In the coagulation process, inorganic coagulants do not compress fibrin strongly, making the structure of the blood clot not tight enough. The state of the blood cells after centrifugation may not be ideal, for example, the blood cells may be loose and flowing, which is not conducive to the detection of high-quality serum.

[0009] May affect test results: If inorganic coagulants are used in certain projects that require extremely high accuracy of test results, their components or some substances produced during the coagulant process may interfere with the test results and affect the doctor's accurate judgment of the patient's condition.

[0010] The coagulant containing thrombin can make 5 ml of venous blood coagulate in 5 to 8 minutes. Although it is relatively faster than inorganic coagulants, it may not be suitable for some scenarios with high requirements for coagulation time, which will affect the efficiency of detection or related operations. Summary of the invention

[0011] In view of the deficiencies in the prior art, the present invention provides a high-efficiency fast-clotting blood coagulant containing thrombin, which solves the problems of the existing coagulants such as slow coagulation speed, poor stability and inability to meet the rapid detection needs of emergency cases.

[0012] To achieve the above objectives, the present invention is implemented by the following technical scheme: a high-efficiency fast-clotting blood coagulant containing thrombin, comprising the following components: Thrombin, 10-60 g; Silicon dioxide, 30-80 g; Inner wall coating agent, 1-20g; Anhydrous ethanol, 1000ml.

[0013] Preferably, the thrombin is lyophilized thrombin.

[0014] Preferably, the particle size of the silicon dioxide is 1-10 μm.

[0015] Preferably, the specific surface area of ​​the silicon dioxide is 100-300m 2 / g.

[0016] Preferably, the specific content of thrombin is 30g.

[0017] Preferably, the specific content of the silicon dioxide is 50g, and the content of the inner wall coating agent is 20g.

[0018] Preferably, the inner wall coating agent is polyvinyl alcohol.

[0019] A method for preparing a high-efficiency fast-clotting blood coagulant containing thrombin comprises the following steps: Step 1: Take 1000 ml of anhydrous ethanol, add it into a stirring container and stir it at a stirring speed of 300-500 rpm for 30-60 minutes; Step 2: Add 30g-80g of silicon dioxide powder into the anhydrous ethanol solution and continue stirring for 30-60 minutes; Step 3, add 10g-60g thrombin and continue stirring for 30-60 minutes; Step 4: Add 1g-20g of inner wall coating agent and stir for 15-30 minutes; Step 5: Filter to obtain a uniform solution and store it Preferably, in steps 1 to 4, the stirring speed is 300-500 rpm, and the stirring time is 30-60 minutes; in step 5, 0.45 μm filter is used for filtering.

[0020] The present invention provides a high-efficiency fast-clotting blood coagulant containing thrombin, which has the following beneficial effects: 1. The present invention uses a high-efficiency fast-clotting blood coagulant containing thrombin, achieving the technical effect of significantly improving the blood coagulation speed. Compared with the technical solution of using inorganic coagulants in the prior art, the present invention can make the blood coagulate rapidly within 3-5 minutes through the direct participation of thrombin, thereby greatly reducing the delay caused by the long coagulation time in clinical and laboratory tests, and improving the overall efficiency of blood sample processing.

[0021] 2. The present invention adopts a composite technical solution of silicon dioxide and thrombin with an optimized ratio, achieving the technical effect of enhancing the serum separation effect. Compared with the technical solution using a single component in the prior art, the composite coagulant of the present invention not only accelerates the blood coagulation process through the synergistic effect of silicon dioxide and thrombin, but also effectively improves the compactness of the blood clot, avoids the precipitation of blood cells and fibrin, ensures the rapid separation of high-quality serum, and provides a reliable sample for subsequent detection and analysis.

[0022] 3. The present invention uses anhydrous ethanol as a technical solution of solvent, achieving the technical effect of improving the activity and stability of thrombin. Compared with the technical solution of using aqueous solution or other organic solvents in the prior art, the present invention utilizes the low water activity of anhydrous ethanol to effectively avoid the problem of loss of thrombin activity or denaturation, thereby ensuring the stability of thrombin during long-term storage. The use of this solvent system ensures the long-term effectiveness and reliability of the coagulant, and further improves the accuracy and consistency of sample processing.

[0023] 4. The present invention adopts the technical solution of inner wall coating agent, which achieves the technical effect of reducing the interaction between blood and the inner wall of the test tube and accelerating the coagulation process. Compared with the technical solution in the prior art that does not treat the inner wall of the test tube or uses non-special coating materials, the present invention effectively reduces the friction and adhesion of the blood sample during the coagulation process by coating the inner wall of the test tube with polyvinyl alcohol film, ensuring that the blood is evenly mixed and coagulated quickly in the test tube. This optimizes the blood coagulation process, avoids the delay in coagulation time caused by poor contact between the blood and the test tube surface, and thus improves the efficiency of the entire detection process. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The figure is a flow chart of the method of the present invention. DETAILED DESCRIPTION

[0025] The following will be combined with the drawings in the specification of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Embodiment 1: Please refer to the attached Figure 1 The embodiment of the present invention provides a high-efficiency rapid coagulation blood coagulant containing thrombin, comprising the following components: Thrombin (30 g); Silicon dioxide (50 g); Inner wall coating agent (20g); Anhydrous ethanol (1000ml).

[0027] This ratio ensures that the coagulation process is rapid and effective while avoiding the adverse effects caused by excessive addition of a certain component. Through a reasonable ratio, thrombin and silica can work together effectively to accelerate the coagulation of blood, while the inner wall coating agent ensures that the blood in the test tube can coagulate smoothly and reduce unnecessary interference. Anhydrous ethanol is used as a solvent to ensure the solubility and stability of each component, so that the entire coagulant can play the best effect when used.

[0028] In this embodiment, the specific technical implementation is as follows: Thrombin (30g) Thrombin plays a core role in the blood coagulation process, and can quickly convert fibrinogen into fibrin to form a blood clot. In order to ensure that the coagulation process is rapid and stable, the present invention uses lyophilized thrombin, which has high enzyme activity, can quickly activate coagulation factors in the blood at room temperature, and promote rapid coagulation of blood.

[0029] Thrombin accelerates the blood coagulation process by catalyzing the conversion of fibrinogen into fibrin. In the present invention, the addition amount of thrombin is 30g, and this ratio ensures the efficient progress of the blood coagulation process. Too little thrombin may cause the coagulation speed to be too slow, while too much may cause the clot structure to be unstable. Therefore, 30g of thrombin has been proven in the present invention to be able to effectively promote the coagulation of blood within 3-5 minutes, and will not affect the compactness and stability of the blood clot.

[0030] Silicon dioxide (50g) As an inorganic coagulant, silicon dioxide accelerates the coagulation process mainly by activating the internal and external coagulation systems of blood. It has good adsorption and a large specific surface area, which can promote the cascade reaction of platelet aggregation and coagulation factors, and accelerate the initial coagulation of blood.

[0031] Silica particles interact with coagulation factors (such as coagulation factors VII and X) in the blood through their surface, activating the coagulation system and starting the coagulation reaction. The particle size (1-10 μm) and specific surface area (100-300 m 2 / g) optimizes its dispersion and contact surface in the blood, effectively promotes the interaction between coagulation factors and platelets, and enables the coagulation process to be initiated quickly. 50g of silicon dioxide in this ratio can ensure its dispersion in the blood sample, avoid precipitation, and maximize its effect of promoting coagulation.

[0032] Inner wall coating agent (20g) The main function of the inner wall coating agent is to coat the inner wall of the test tube to reduce the friction and adhesion between the blood and the test tube surface, and prevent the friction or adhesion from interfering with the coagulation process. The common inner wall coating agent is polyvinyl alcohol, which can form a uniform film to prevent the inner wall of the test tube from adsorbing the blood sample.

[0033] The inner wall coating agent forms a thin film on the inner wall of the test tube, reducing the friction and adhesion between the blood and the test tube surface, allowing the blood sample to flow freely, thereby accelerating the coagulation reaction. This layer of film reduces the interaction between the components in the blood and the test tube surface, avoiding unnecessary losses and interference, and ensuring that the blood can complete coagulation and form a stable blood clot in a shorter time. 20g of the inner wall coating agent can effectively cover the entire inner wall of the test tube at this ratio, ensuring a smooth coagulation process.

[0034] Anhydrous ethanol (1000ml) As a solvent, anhydrous ethanol can help dissolve and disperse other ingredients, ensure the uniform distribution of each component, and maintain its activity. Ethanol can reduce the water activity in the solution, avoid the inactivation or degradation of thrombin due to water, and thus maintain its catalytic efficiency.

[0035] Anhydrous ethanol prevents the decrease of thrombin activity by reducing the water activity in the solution and helps other components to be evenly dispersed in the solution. Ethanol can also promote the stable dissolution of thrombin and other components such as silicon dioxide, thereby ensuring that they can function effectively when used. 1000ml of anhydrous ethanol ensures the stability of the solution and can effectively dissolve and disperse all components, providing a stable reaction environment.

[0036] Based on the above-mentioned ratio content, this embodiment also discloses a method for preparing a high-efficiency fast-clotting blood coagulant containing thrombin, comprising the following steps: Step 1: Solvent preparation: First, measure 1000 ml of anhydrous ethanol and add it to a beaker, and make sure that the ethanol is free of water contamination. Place the beaker on a magnetic stirrer and start stirring.

[0037] Step 2: Mix the ingredients. During the stirring process, slowly add 30g of thrombin and ensure that it is fully dissolved. Then add 50g of silicon dioxide and continue stirring until the silicon dioxide is evenly dispersed.

[0038] Step 3: Add inner wall coating agent. Add 20g of inner wall coating agent to ensure that the inner wall coating agent is evenly dispersed in the solution to avoid precipitation.

[0039] Step 4: Adjust the pH value of the solution. Through appropriate pH adjustment, ensure that the solution is in the neutral to weakly alkaline range to ensure the activity of thrombin.

[0040] Step 5: Packaging and storage: Pack the evenly mixed solution into appropriate containers, seal them and store them in a cool and dry place, avoiding exposure to moisture or high temperature.

[0041] Example 2: Low-concentration adaptive thrombin-containing high-efficiency fast-clotting blood coagulant step: Prepare the ingredients Thrombin: 15 g Silicon dioxide: 60g Inner wall coating agent (polyvinyl alcohol): 15g Anhydrous ethanol: 1000ml Dissolving and mixing: Add 1000 ml of anhydrous ethanol into a container, add 60 g of silicon dioxide, and stir until the silicon dioxide is completely dissolved.

[0042] Add 15 g of polyvinyl alcohol and continue stirring until it is completely dissolved.

[0043] Add thrombin: Add 15 g of thrombin to the solution and stir until the thrombin is completely dissolved, ensuring even distribution.

[0044] Inspection and adjustment: Test the coagulation time under standard conditions to ensure that coagulation is completed within 5 minutes. Check whether the contraction time of the blood clot meets the standard.

[0045] Packaging and storage: Dispense the coagulant into test tubes and store under refrigerated conditions.

[0046] Process parameters: Temperature: 25℃ (room temperature) Silica particle size: 2-8 μm, polyvinyl alcohol concentration: 15 g, thrombin addition amount: 15 g, stirring time: 25 minutes, storage conditions: refrigerated Summary of embodiments: This embodiment adopts the ratio of 15g thrombin, 60g silicon dioxide, and 15g inner wall coating agent, which is suitable for environments with loose requirements for coagulation time. By reducing the amount of thrombin added, it is ensured that the coagulation process will not be too fast within a long time window, and the blood clot structure is more stable. This ratio is more suitable for clinical applications that require a long time to process or a large sample size.

[0047] This embodiment is suitable for scenarios that require a slower coagulation speed. Reducing the amount of thrombin can effectively prolong the coagulation time, avoiding premature formation of unstable blood clots in blood samples due to excessively rapid coagulation in certain cases, thereby affecting subsequent centrifugation and sample processing. At the same time, increasing the proportion of silicon dioxide helps to improve the stability and uniformity of the coagulation process. It is suitable for some routine tests, especially for scenarios with high accuracy requirements.

[0048] Example 3: Highly effective and fast-clotting blood coagulant containing thrombin (applicable to a large number of samples) step: Prepare the raw materials: 50g thrombin, 40g silicon dioxide, 10g inner wall coating agent (polyvinyl alcohol), 1500ml anhydrous ethanol Dissolving and mixing: Add 1500ml of anhydrous ethanol to a beaker, add 40g of silicon dioxide, and gently stir until the silicon dioxide is completely dissolved. Add 10g of the inner wall coating agent and stir until it is completely dissolved.

[0049] Add thrombin: Add 50 g of thrombin to the above solution and continue stirring until the thrombin is completely dissolved, ensuring that all components in the solution are evenly distributed.

[0050] Inspection and adjustment: At room temperature, take a small amount of the mixed solution for coagulation time and clot retraction time testing to confirm that it meets the coagulation time standard of ≤5 minutes and clot retraction time of ≤15 minutes.

[0051] Packaging and storage: Divide the prepared coagulant solution into sealed test tubes and store them under refrigerated conditions to ensure that the product is not inactivated during storage.

[0052] Process parameters: Temperature: 25℃ (room temperature) Silica particle size: 1-10μm Polyvinyl alcohol concentration: 10 g, thrombin addition amount: 50 g, stirring time: 40 minutes, storage conditions: refrigerated Summary of embodiments: This embodiment adopts a ratio of 50g thrombin, 40g silicon dioxide, and 10g inner wall coating agent, which can effectively meet the needs of large-scale sample processing. This ratio can quickly and evenly promote blood coagulation, is suitable for the detection of large batches of samples, ensures that the coagulation time is within 5 minutes, and the blood clot contraction time is controlled within 15 minutes.

[0053] The thrombin content of this embodiment is higher, which helps to ensure a rapid and efficient coagulation reaction in the process of processing large quantities of samples. In large sample or emergency scenarios, the increase in the amount of thrombin can ensure a more rapid coagulation effect to improve the work efficiency of the laboratory. Compared with other embodiments, the higher thrombin ratio can effectively reduce waiting time and ensure the timeliness of sample processing, especially when processing a large number of samples that urgently need rapid diagnosis, it can achieve higher work efficiency.

[0054] Example 4: Highly effective and stable thrombin-containing high-efficiency and fast-clotting blood coagulant step: Prepare the raw materials: thrombin: 35g, silicon dioxide: 55g, inner wall coating agent (polyvinyl alcohol): 10g, anhydrous ethanol: 1000ml Dissolving and mixing: Add 1000 ml of anhydrous ethanol to a beaker, add 55 g of silica, and stir thoroughly to ensure it is evenly dispersed.

[0055] Add 10g of inner wall coating agent and continue stirring to ensure complete dissolution.

[0056] Add thrombin: Add 35 g of thrombin to the solution and stir until the thrombin is completely dissolved.

[0057] Inspection and adjustment: Take a small amount of the mixture and perform a coagulation time test to ensure that it is completely coagulated within 4 minutes. Check the contraction time of the blood clot and confirm that it is ≤12 minutes.

[0058] Packaging and Storage: Divide the mixture into test tubes and store them in a refrigerator to maintain their stability.

[0059] Process parameters: Temperature: 25°C (room temperature), silica particle size: 3-10 μm, polyvinyl alcohol concentration: 10 g, thrombin addition amount: 35 g, stirring time: 30 minutes, storage conditions: refrigerated Summary of embodiments: This embodiment adopts a ratio of 35g thrombin, 55g silicon dioxide, and 10g inner wall coating agent, which is suitable for rapid and stable blood coagulation. By reasonably adjusting the concentration of thrombin, it is ensured that the blood can coagulate within 4 minutes, and the stability of blood clot contraction is guaranteed.

[0060] Compared with Example 1, the amount of thrombin added is moderately reduced, so that the speed of the coagulation reaction is slightly slowed down, ensuring that the blood clot structure after coagulation is more compact. The proportion of silicon dioxide is increased, providing sufficient stability, making this ratio particularly suitable for situations where there is a certain tolerance in time, ensuring that the effect of rapid coagulation can be achieved while not affecting the final quality of the blood clot.

[0061] Comparative Example 1: Using a lower concentration of thrombin (Comparative Example 2) Prior art (Comparative Example 1): Ratio: Thrombin: 10g, Silicon dioxide: 60g, Inner wall coating agent: 15g, Anhydrous ethanol: 1000ml Experimental process: Raw material preparation: Prepare the components according to the above proportions, ensuring that the silica and inner wall coating agent are completely dissolved in ethanol.

[0062] Mixing: Add thrombin in order and stir evenly.

[0063] Test: Test the coagulation time and clot retraction time. The expected coagulation time is less than 5 minutes and the clot retraction time is less than 15 minutes.

[0064] Comparative Example: Example 2: Using a ratio of 15 g thrombin, 60 g silicon dioxide, and 15 g inner wall coating agent, compared with comparative example 1, the thrombin concentration is moderately increased to ensure a faster and more stable coagulation reaction rate.

[0065] Comparative analysis: Coagulation time: In comparative example 1, the lower thrombin concentration (10 g) resulted in a relatively long coagulation time, usually more than 5 minutes, which is suitable for some scenarios where the coagulation speed is not required. In the embodiment of the present invention, the thrombin concentration is higher (15 g), which makes the coagulation time faster, meeting the clinical rapid detection requirements.

[0066] Blood clot structure: In comparative example 1, due to the low concentration of thrombin, the blood clot structure may be loose and difficult to form a tight blood clot, which has a great impact on the quality of centrifugation and serum extraction. The embodiment of the present invention ensures that the concentration of thrombin can accelerate the shrinkage of blood clots, improve the blood processing efficiency and subsequent detection accuracy.

[0067] Comparative Example 2: Using a ratio without an inner wall coating agent (Comparative Example 1) Prior art (Comparative Example 2): Ratio: Thrombin: 30g, Silicon dioxide: 50g, Anhydrous ethanol: 1000ml Experimental process: Raw material preparation: prepare thrombin, silicon dioxide and ethanol according to the ratio.

[0068] Mixing: Mix thrombin with silica and add ethanol to dissolve.

[0069] Tests: Measures the clotting time and clot retraction time.

[0070] Comparative Example: Example 1: Contains 20 g of inner wall coating agent (polyvinyl alcohol) to improve the stability of blood clots and reduce interference with blood samples.

[0071] Comparative analysis: Coagulation reaction stability: In comparative example 2, there is no inner wall coating agent. Although the coagulation time is achieved within 3-5 minutes, due to the lack of the protective effect of the coating film, the interaction between the coagulation substance and the container wall may affect the purity of the blood sample during sample processing. In contrast, the present invention ensures the stability of the blood coagulation process by using the inner wall coating agent, avoiding secondary contamination and interference.

[0072] Blood clot stability: The embodiments of the present invention effectively reduce the adhesion of blood clots to the container wall by using an inner wall coating agent, thereby ensuring the stability of the blood clot after coagulation and a good serum separation effect.

[0073] Comparative Example 3: Proportion without silicon dioxide (Comparative Example 3) Prior art (Comparative Example 3): Ratio: Thrombin: 40g, inner wall coating agent: 15g, anhydrous ethanol: 1000ml Experimental process: Raw material preparation: prepare thrombin, inner wall coating agent and ethanol according to the ratio.

[0074] Mixing: After mixing thrombin with the inner wall coating agent, add ethanol to dissolve.

[0075] Tests: Measures the clotting time and clot retraction time.

[0076] Comparative Example: Example 3: In the example of the present invention, silicon dioxide is added to make the coagulation process faster and more stable by utilizing its property of promoting the reaction of the coagulation system.

[0077] Comparative analysis: Coagulation efficiency: In comparative example 3, the effect of silicon dioxide is lacking. Although the concentration of thrombin is high, the coagulation time may be disturbed, especially under the requirement of lower coagulation speed, the blood coagulation speed is still not ideal. Silicon dioxide can better activate the blood coagulation system, especially has a good effect on the activation of coagulation factors. On this basis, the embodiment of the present invention uses silicon dioxide to enhance the efficiency of the coagulant.

[0078] Blood clot firmness: A coagulant without silicon dioxide may cause the blood clot to be loose and not compact, thus affecting the separation effect of serum. In contrast, the embodiments of the present invention improve the structural stability of the blood clot by adding silicon dioxide in a reasonable manner, thereby achieving a better serum separation effect.

[0079] Comparative Example 4: Increasing the amount of inner wall coating agent (Comparative Example 2) Prior art (Comparative Example 4): Ratio: Thrombin: 15g, Silicon dioxide: 60g, Inner wall coating agent: 30g, Anhydrous ethanol: 1000ml Experimental process: Preparation of raw materials: prepare thrombin, silicon dioxide, inner wall coating agent and ethanol according to the above proportions.

[0080] Mixing: Add all ingredients to ethanol and stir until completely dissolved.

[0081] Inspection: Test the coagulation time and clot retraction time to ensure that they meet the standards.

[0082] Comparative Example: Example 2: Contains 15 g of inner wall coating agent. A smaller amount of coating agent ensures the balance of the coagulation reaction.

[0083] Comparative analysis: Coagulation time: In comparative example 4, due to the high concentration of the inner wall coating agent (30 g), the coagulation process may be too slow or too stable, affecting the demand for rapid blood coagulation. Excessive inner wall coating agent may cause the reaction to be too slow and the blood coagulation time to be prolonged, which is not suitable for scenarios requiring rapid detection.

[0084] Blood clot stability: Although the addition of the inner wall coating agent helps to improve the stability of the blood clot, excessive use may cause the blood clot to be too hard, affecting the subsequent centrifugation operation and even the serum separation effect. In the embodiment of the present invention, the amount of the inner wall coating agent used is 20g, which can maintain the stability of the blood clot without excessively affecting the coagulation speed.

[0085] Experiment 1: Coagulation time comparison experiment Experimental purpose: This experiment aims to verify the effects of different combinations of coagulants on blood coagulation time, especially the effect of the coagulant of the present invention on accelerating blood coagulation. By comparing the coagulation time of the embodiment of the present invention with that of the comparative example, the acceleration effect of different composition ratios and coagulant types on coagulation time is verified.

[0086] Experimental steps: Sample preparation: Prepare venous blood samples, each sample volume is 2ml. Make sure that the blood samples are collected at the same time period and try to avoid interference from external factors after collection.

[0087] Preparation of coagulant: Prepare different coagulants separately according to the ratios of the experimental group and the comparative group.

[0088] Experimental group (Example 1): Proportion: 30 g of thrombin, 50 g of silicon dioxide, 20 g of inner wall coating agent, and 1000 ml of anhydrous ethanol.

[0089] Comparative Example 1: Proportion: 20g thrombin, 50g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0090] Comparative Example 2: Proportion: 30g thrombin, 60g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0091] Mix and Observe: Dispense each coagulant group into labeled tubes, labeling them clearly.

[0092] Add 2 ml of blood sample to each test tube.

[0093] Gently invert the tube 5-10 times to thoroughly mix the blood and coagulant.

[0094] Place the test tube at room temperature (25°C) and observe and record the time required for blood clotting.

[0095] Record and statistics: Observe the blood coagulation time from the addition of the coagulant to the complete coagulation of the blood. Repeat each group of experiments 3 times, record the coagulation time each time, and calculate the average value.

[0096] Table 1: Coagulation time test data From the results of this experiment, it can be seen that Example 1 (i.e., the solution of the present invention) is significantly better than the coagulation time of Comparative Example 1 and Comparative Example 2. The average coagulation time of Example 1 is 3.26 minutes, while the average coagulation time of Comparative Example 1 is 4.2 minutes, and the average coagulation time of Comparative Example 2 is 3.87 minutes. The experimental data show that the coagulation time acceleration effect of the present invention is more significant, which can greatly shorten the blood coagulation time, especially in emergency and application scenarios where samples need to be processed quickly, and can effectively improve the overall efficiency.

[0097] The coagulant of the present invention comprises a combination of thrombin and silicon dioxide, which plays a key role in accelerating the blood coagulation process. As a biologically active ingredient, thrombin can quickly convert the fibrinogen in the blood into fibrin, and then form a blood clot, and this process requires a shorter time. In this experiment, the ratio of 30g thrombin and 50g silicon dioxide in Example 1 can not only activate the coagulation reaction more quickly, but also enhance the role of silicon dioxide particles as a surfactant in accelerating the coagulation process. Silicon dioxide can activate the internal and external coagulation mechanism of blood and improve the efficiency of coagulation reaction. Therefore, the present invention can complete coagulation in just a few minutes, which is significantly better than traditional inorganic coagulants.

[0098] Compared with Comparative Example 1 (lower thrombin content) and Comparative Example 2 (using more silica), the balanced ratio in Example 1 enables the coagulant to more effectively activate the coagulation reaction. Comparative Example 1 has a slower coagulation reaction start-up speed due to less thrombin, and although Comparative Example 2 increases the amount of silica, the excess silica may cause too much interference to the coagulation process, making the coagulation reaction not rapid and accurate enough. Therefore, the optimal ratio in Example 1 provides the best coagulation acceleration effect, can effectively shorten the blood coagulation time, and ensure more efficient separation of serum or plasma for subsequent testing in clinical practice.

[0099] Experiment 2: Comparative experiment on blood clot contraction time Experimental purpose: This experiment aims to verify the effect of different coagulant combinations on blood clot contraction time. By comparing the blood clot contraction time of the embodiment of the present invention with that of the comparative example, the effect of the coagulant of the present invention in accelerating blood clot contraction after promoting blood coagulation is demonstrated, and its advantages in practical applications are further verified, especially its beneficial effect in improving the quality of blood samples.

[0100] Experimental steps: Sample preparation: Prepare venous blood samples, the collection volume is 2ml. Ensure that the blood sample collection process is strictly controlled to reduce external factors that may affect the experimental results.

[0101] Preparation of coagulant: Prepare different coagulants according to the ratios of the experimental group and the control group.

[0102] Experimental group (Example 1): Proportion: 30 g of thrombin, 50 g of silicon dioxide, 20 g of inner wall coating agent, and 1000 ml of anhydrous ethanol.

[0103] Comparative Example 1: Proportion: 20g thrombin, 50g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0104] Comparative Example 2: Proportion: 30g thrombin, 60g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0105] Mix and observe: Add each coagulant according to the above proportions into the marked test tubes.

[0106] Add 2 ml of blood sample to each test tube, making sure that the blood sample and coagulant in each test tube are fully mixed.

[0107] Gently invert the tube 5-10 times to mix the blood and coagulant evenly.

[0108] The test tube was placed at room temperature (22-25°C).

[0109] The shrinkage of the blood clot was observed at fixed time intervals (e.g., every 1 minute), and the time when serum began to precipitate was recorded.

[0110] The experiment was repeated three times in each group, the time of each blood clot contraction was recorded, and the average value was calculated.

[0111] Table 2: Experimental data of blood clot contraction time Experimental summary: From the experimental data, it can be seen that Example 1 (the solution of the present invention) shows a significant advantage in blood clot contraction time. The average blood clot contraction time of Example 1 is 12.43 minutes, while the average time of Comparative Example 1 is 15.17 minutes, and the average time of Comparative Example 2 is 13.53 minutes. It can be clearly seen from the comparison that the present invention can significantly shorten the blood clot contraction time and further enhance the processing efficiency of blood samples. Especially for the clinical environment of rapid processing and analysis of samples, the present invention has important practical application value.

[0112] The present invention significantly accelerates the formation of fibrin in the blood and the tightness of its network structure through an effective combination of coagulants. While promoting blood coagulation, thrombin also accelerates the interweaving and network formation of fibrin. After the fibrin network is formed, it will promote the contraction of blood clots and separate serum from blood cells. In Example 1, the appropriate ratio of thrombin concentration and silicon dioxide makes the network structure of fibrin tighter during the coagulation process, thereby promoting the contraction process of blood clots. In contrast, the thrombin concentration in Comparative Example 1 is low, resulting in a slower contraction speed of the blood clot; the content of silicon dioxide in Comparative Example 2 is high, but too much silicon dioxide may not enhance its effect, but may lead to over-activation in the coagulation process, affecting the tightness of the blood clot, thereby prolonging the contraction time of the blood clot.

[0113] The mechanism analysis of this experiment can further prove that the coagulant of the present invention not only accelerates the coagulation process of blood, but also optimizes the effect of blood clot contraction through a more efficient ratio of silica and thrombin, avoids the side effects caused by over-activation of the coagulation system, and ultimately improves the quality and processing efficiency of blood samples.

[0114] Experiment 3: Comparative experiment on serum separation efficiency Experimental purpose: This experiment aims to further verify that the present invention can effectively improve the quality of serum separation while accelerating blood sample processing by comparing the effects of different coagulant combinations on serum separation efficiency, and provide strong technical support for rapid sample processing in clinical experiments and scientific research.

[0115] Experimental steps: Sample preparation: Collect venous blood samples from healthy adults, 2 ml for each sample. When collecting all blood samples, ensure that there is no external contamination and collect them under the same environmental conditions.

[0116] Preparation of coagulant: According to the ratios of the experimental group and the comparative example, different coagulants were prepared respectively, and the specific ratios were as follows: Experimental group (Example 1): Ratio: 30 g thrombin, 50 g silicon dioxide, 20 g inner wall coating agent, 1000 ml anhydrous ethanol.

[0117] Comparative Example 1: Proportion: 20g thrombin, 50g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0118] Comparative Example 2: Proportion: 30g thrombin, 60g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0119] Mix and Observe: Add each coagulant into the test tube according to the above ratio to ensure the correct amount of coagulant in each test tube.

[0120] Add the collected blood sample into each test tube as required to ensure that the blood sample and the coagulant are evenly mixed.

[0121] Gently invert the test tube 5-10 times to allow the blood to fully contact with the coagulant.

[0122] Place the test tube at room temperature (22-25°C), observe whether the blood completely coagulates in a short period of time, and record the time when coagulation begins.

[0123] After the blood was completely coagulated, it was centrifuged (3000 rpm, 10 minutes) and the serum separation was observed and recorded.

[0124] Record the transparency of the serum, the compactness of the blood clot, and the amount of serum in each test tube.

[0125] Data collection and recording: Record the clarity of each group of sample serum (1-5 points, 1 point for turbidity, 5 points for transparency), as well as the volume of separated serum (unit: ml). Each group of experiments was repeated three times, and the average value was calculated.

[0126] Table 3: Experimental data of serum separation efficiency It can be seen from the experimental data that the serum separation effect of Example 1 of the present invention is significantly better than that of the comparative example group. Example 1 has achieved better results in serum clarity (5 points), serum volume (more than 1.5 ml), blood clot tightness (5 points) and serum separation time (about 8 minutes). Compared with Comparative Example 1 (serum clarity 4 points, less serum volume, and longer serum separation time), Example 1 can significantly improve the separation efficiency of serum, effectively improve the quality of blood samples, and reduce the processing time of samples. The effect of Comparative Example 2 is closer to that of Example 1, but still inferior to that of Example 1.

[0127] The reasonable ratio of thrombin and silicon dioxide of the present invention plays a key role in the coagulation process. The addition of thrombin promotes the rapid conversion of fibrinogen in the blood into fibrin, forming a tight clot structure. Silicon dioxide, as a solid particle, can accelerate the blood coagulation process and help fibrin to be interwoven into a network structure, thereby promoting the contraction of the blood clot. In Example 1, the moderate amount of thrombin and the optimal ratio of silicon dioxide can give full play to the effects of these two components, not only shortening the coagulation time, but also ensuring the high efficiency of blood clot contraction, so that serum separation is faster and clearer. However, the thrombin content of comparative example 1 is too low, resulting in an insufficiently tight clot structure, a small amount of separated serum and poor quality; although comparative example 2 increases the content of silicon dioxide, its effect is still not as good as that of Example 1 of the present invention, indicating that a single increase in the content of silicon dioxide cannot effectively improve the serum separation effect, but may lead to a less than ideal coagulation process, affecting the quality of the blood sample.

[0128] Experiment 4: Experiment on the effect of different coagulants on blood clotting time Experimental purpose: This experiment aims to verify the effect of the high-efficiency fast-clotting blood coagulant provided by the present invention in accelerating the blood clotting process by comparing the blood clotting time of different coagulant combinations, focusing on observing the specific effect of the ratio of thrombin and silica on the clotting time.

[0129] Experimental steps: Sample preparation: Collect venous blood samples from healthy adults, 2 ml for each sample. When collecting all blood samples, ensure that there is no external contamination and collect them under the same environmental conditions.

[0130] Preparation of coagulant: different coagulants were prepared according to the ratio of the experimental group and the comparative example. The specific ratios are as follows: Experimental group (Example 1): Proportion: 30 g of thrombin, 50 g of silicon dioxide, 20 g of inner wall coating agent, and 1000 ml of anhydrous ethanol.

[0131] Comparative Example 1: Proportion: 20g thrombin, 50g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0132] Comparative Example 2: Proportion: 30g thrombin, 60g silicon dioxide, 20g inner wall coating agent, 1000ml anhydrous ethanol.

[0133] Comparative Example 3: Ratio: 25g thrombin, 50g silicon dioxide, 15g inner wall coating agent, 1000ml anhydrous ethanol.

[0134] Mix and Observe: Add each coagulant into the test tube according to the above ratio to ensure the correct amount of coagulant in each test tube.

[0135] Add the collected blood sample into each test tube as required to ensure that the blood sample and the coagulant are evenly mixed.

[0136] Gently invert the tube 5-10 times to ensure that the coagulant is in full contact with the blood.

[0137] The test tube was placed at room temperature (22-25°C) and the time when the blood began to coagulate was recorded.

[0138] The blood coagulation progress was checked every 30 seconds, and the total time required for blood coagulation was recorded.

[0139] Data collection and recording: Record the time when each group of samples starts to coagulate and the time when they are completely coagulated. Repeat each group of experiments three times and calculate the average value.

[0140] Table 4: Effects of different coagulants on blood coagulation time From the experimental data, Example 1 is significantly better than other comparative example groups. In terms of coagulation start time and complete coagulation time, Example 1 performs best, and the coagulation time difference (i.e., the time from the addition of the coagulant to complete coagulation of the blood) is significantly lower than that of Comparative Example 1, Comparative Example 2 and Comparative Example 3. The coagulation time of Example 1 is shorter, and the coagulation process is more efficient, indicating that the optimal ratio of thrombin and silicon dioxide can significantly accelerate the blood coagulation process. In contrast, the coagulation time difference of Comparative Example 1 and Comparative Example 3 is longer, indicating that the excess of silicon dioxide or the deficiency of thrombin may affect the activity of the coagulant and the coagulation efficiency.

[0141] The role of thrombin in the present invention is to catalyze the conversion of fibrinogen into fibrin, form a tight clot structure, and accelerate the process of blood coagulation. As a solid particulate matter, silicon dioxide can not only promote the rapid reaction of thrombin and blood, but also increase the activity of coagulation factors in the coagulation process and shorten the coagulation time. The optimal ratio of thrombin and silicon dioxide in Example 1 can effectively regulate the interaction between fibrinogen and coagulation factors in the blood, thereby achieving a more efficient and faster coagulation process. On the contrary, if the ratio of thrombin is too low or there is too much silicon dioxide, it may lead to prolonged coagulation time and the coagulation process is not smooth enough. Therefore, the high-efficiency and fast-clotting blood coagulant of the present invention has significant advantages in blood sample processing, and is particularly suitable for clinical scenarios where rapid detection is required.

[0142] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A high-efficiency fast-clotting blood coagulant containing thrombin, characterized in that: Includes the following components: Thrombin, 10-60 g; Silicon dioxide, 30-80 g; Inner wall coating agent, 1-20g; Anhydrous ethanol, 1000ml.

2. The high-efficiency rapid-coagulation blood coagulant containing thrombin according to claim 1, characterized in that: The thrombin is freeze-dried thrombin.

3. The high-efficiency rapid-coagulation blood coagulant containing thrombin according to claim 1, characterized in that: The particle size of the silicon dioxide is 1-10 μm.

4. The high-efficiency rapid-coagulation blood coagulant containing thrombin according to claim 1, characterized in that: The specific surface area of ​​the silicon dioxide is 100-300m ² / g.

5. The high-efficiency rapid-coagulation blood coagulant containing thrombin according to claim 1, characterized in that: The specific content of the thrombin is 30g.

6. The high-efficiency rapid-coagulation blood coagulant containing thrombin according to claim 1, characterized in that: The specific content of the silicon dioxide is 50g, and the content of the inner wall coating agent is 20g.

7. The high-efficiency rapid-coagulation blood coagulant containing thrombin according to claim 1, characterized in that: The inner wall coating agent is polyvinyl alcohol.

8. A method for preparing a high-efficiency and fast-clotting blood coagulant containing thrombin, according to the high-efficiency and fast-clotting blood coagulant containing thrombin according to any one of claims 1 to 6, characterized in that: The following steps are involved: Step 1: Take 1000 ml of anhydrous ethanol, add it into a stirring container and stir it at a stirring speed of 300-500 rpm for 30-60 minutes; Step 2: Add 30g-80g of silicon dioxide powder into the anhydrous ethanol solution and continue stirring for 30-60 minutes; Step 3: Add 10g-60g thrombin and continue stirring for 30-60 minutes; Step 4: Add 1g-20g of inner wall coating agent and stir for 15-30 minutes; Step 5: Filter to obtain a uniform solution and store it.

9. The method for preparing a high-efficiency rapid-coagulation blood coagulant containing thrombin according to claim 8, characterized in that: In step 1 to step 4, the stirring speed is 300-500 rpm, and the stirring time is 30-60 minutes; in step 5, a 0.45 μm filter is used for filtering.

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