Collagen material, its preparation method and application in collagen hemostatic powder

Abstract

The application relates to the technical field of collagen materials, and particularly discloses a collagen material, a preparation method thereof and application of the collagen material in collagen hemostatic powder. The preparation method of the collagen material is as follows: enzyme hydrolysis reaction: adjusting a raw material sample liquid to pH 3.5-6.0, adding a composite enzyme, carrying out enzyme hydrolysis at a temperature of 30-40 DEG C for 4-12 h, and centrifuging to obtain a supernatant; the composite enzyme is composed of trypsin and phospholipase in a weight ratio of 1-5:0.02-0.08; alkali treatment: adjusting the supernatant to pH>13, and carrying out alkali treatment at a temperature of 15-25 DEG C for more than 40 h; neutralization reaction: adjusting pH to 6.8-7.6, eluting, and removing water by extrusion, so that the collagen material is obtained. The application also provides application of the collagen material in the collagen hemostatic powder. The collagen hemostatic powder provided by the application has excellent hemostatic function and water absorption efficiency, and the hemostatic material has high stability.

Description

Technical Field

[0001] This application relates to the technical field of adhesive raw materials, specifically to an adhesive raw material, its preparation method, and its application in collagen hemostatic powder. Background Technology

[0002] Collagen raw materials have garnered widespread attention in the medical field due to their excellent hemostatic properties and ability to guide tissue repair and regeneration. However, while the chemical cross-linking agents used in traditional processes improve material stability, they also introduce complex manufacturing processes and potential safety hazards, leading to residual cross-linking agents potentially causing cytotoxicity and adverse immune responses. Furthermore, existing collagen extraction and purification techniques often fail to effectively remove impurities, such as residual fat, which can also affect the biocompatibility and clinical safety of the materials. Summary of the Invention

[0003] To address the aforementioned technical problems, this application provides a collagen raw material, its preparation method, and its application in collagen hemostatic powder.

[0004] In a first aspect, this application provides a method for preparing an adhesive raw material, specifically comprising the following steps in sequence:

[0005] Sample solution: Take the raw material, freeze and slice it, and place it in 8-15 times its weight of water to make the sample solution;

[0006] Enzymatic hydrolysis: Adjust the pH of the sample solution to 3.5-6.0, add the compound enzyme, and hydrolyze at 30-40℃ for 4-12 hours. Centrifuge and collect the supernatant. The weight ratio of the compound enzyme to the raw material is 1000:0.04-0.12. The compound enzyme is composed of trypsin and phospholipase in a weight ratio of 1-5:0.02-0.08.

[0007] Alkalization reaction: Adjust the pH of the supernatant to >13 and alkalize it at 15-25℃ for more than 40 hours;

[0008] Neutralization reaction: Adjust the pH to 6.8-7.6, elute, squeeze out water to obtain the rubber raw material.

[0009] This application employs a combination of enzymatic hydrolysis, alkalization, and neutralization reactions. A complex enzyme consisting of trypsin and phospholipase in a weight ratio of 1-5:0.02-0.08 is used for enzymatic hydrolysis, and the pH value during the hydrolysis process is controlled. Simultaneously, the process parameters for the alkalization and neutralization reactions are optimized to prepare a collagen raw material with high extraction rate, purity, and biosafety. Furthermore, this collagen raw material exhibits excellent application results in the preparation of collagen-based composite hemostatic powder.

[0010] In the enzymatic hydrolysis stage, the pH of the sample solution is precisely adjusted to 3.5-6.0, and a specific ratio of a compound enzyme is added. This compound enzyme, composed of trypsin and phospholipase in an optimized ratio, can efficiently hydrolyze the sample under mild conditions of 30-40℃. After 4-12 hours of enzymatic hydrolysis, the supernatant is obtained by centrifugation. The key to this step is to ensure the full progress of the enzymatic hydrolysis reaction, thereby maximizing the release of the target gum raw material. Subsequently, the alkalization reaction stage begins. The pH of the supernatant is adjusted to greater than 13, and alkalization is carried out at a low temperature of 15-25℃ for more than 40 hours. This step effectively removes impurities while maintaining the integrity and bioactivity of the gum raw material. The long alkalization process ensures the thorough removal of impurities. Then, in the neutralization reaction stage, the pH is again precisely adjusted to 6.8-7.6. Through elution and dehydration by squeezing, high-purity, highly biosafe gum raw material is finally obtained. The key to this step is to ensure that the pH value of the gum raw material is moderate, neither affecting its bioactivity nor hindering subsequent application and processing. Based on the above, by precisely controlling the conditions and time of enzymatic hydrolysis, alkalization and neutralization reactions, this application has successfully developed a highly efficient, high-purity and biosafety-based method for extracting gum raw materials.

[0011] Preferably, the process parameters for the enzymatic hydrolysis reaction are: enzymatic hydrolysis at 33-38℃ for 8-10 hours.

[0012] Preferably, the weight ratio of the compound enzyme to the bovine Achilles tendon raw material is 1000:0.06-0.10.

[0013] Preferably, the complex enzyme is composed of trypsin and phospholipase in a weight ratio of 2-4:0.04-0.06.

[0014] Preferably, the raw material is selected from one or more of mammalian Achilles tendons and mammalian surface skin.

[0015] Secondly, this application provides an adhesive raw material, which is prepared using the above-described preparation method.

[0016] Thirdly, this application provides a collagen hemostatic powder, which is obtained by freeze-drying and grinding the above-mentioned collagen raw materials into powder;

[0017] Alternatively, the above-mentioned adhesive raw materials can be prepared into a flocculent liquid, freeze-dried and ground into powder;

[0018] Alternatively, the above-mentioned raw materials can be formulated into a flocculent liquid, and additives can be added to prepare a collagen-based composite hemostatic powder.

[0019] Fourthly, this application provides a collagen-based composite hemostatic powder, specifically comprising the following components: 12-16 parts of collagen raw material, 4-8 parts of modified starch, 0.6-1.4 parts of polyethyleneimine, and 60-80 parts of 10-20 wt% aqueous acetic acid solution;

[0020] The modified starch is prepared by dispersing starch and corn oligopeptides in a 5-15 wt% boric acid aqueous solution, mixing and stirring at 40-60℃ for 60-90 min, and then drying and grinding to obtain the modified starch.

[0021] In the technical solution provided in this application, the gelatinous raw material, as the main component of the hemostatic material, possesses excellent biocompatibility and biodegradability. It can quickly adhere to the bleeding site, forming a temporary physical barrier to prevent further blood loss. Simultaneously, the porous structure of the gelatinous raw material facilitates the aggregation of platelets and coagulation factors, accelerating the coagulation process. Secondly, the addition of modified starch provides the hemostatic material with better water absorption and swelling properties; it can rapidly absorb water from the blood, causing the material to swell and tightly fill the bleeding site, thus more effectively applying pressure for hemostasis. Furthermore, modified starch can form a stable complex with the gelatinous raw material, enhancing the material's mechanical strength and stability. The introduction of polyethyleneimine brings a positive charge effect to the collagen hemostatic powder; since red blood cells carry a negative charge on their surface, polyethyleneimine can rapidly aggregate red blood cells to the bleeding site through electrostatic attraction, forming a tight thrombus, thereby achieving rapid hemostasis. Simultaneously, polyethyleneimine also has antibacterial properties, effectively preventing infection and creating a favorable environment for wound healing. Therefore, collagen raw materials, modified starch, and polyethyleneimine, as raw materials for collagen-based composite hemostatic powder, achieve rapid, effective, and safe hemostatic effects through their unique functions and synergistic effects. At the same time, the hemostatic material prepared by this method has excellent stability and a longer shelf life. This innovative hemostatic material is expected to play an important role in the medical field and bring benefits to patients' treatment.

[0022] The modified starch in the adhesive raw material is reacted with starch, corn oligopeptides, and boric acid aqueous solution under specific temperature and time control, which optimizes the microstructure of the modified starch, promotes platelet adhesion and aggregation, and improves its compatibility with blood and hemostatic effect. Simultaneously, the dried and ground modified starch is more likely to synergistically interact with the adhesive raw material and polyethyleneimine, thus adhering closely to the bleeding site and further enhancing the hemostatic effect.

[0023] Preferably, the collagen-based composite hemostatic powder is prepared from the following components in parts by weight: 13-15 parts of collagen raw material, 5-7 parts of modified starch, 0.8-1.2 parts of polyethyleneimine, and 65-75 parts of 10-20 wt% aqueous acetic acid solution.

[0024] Preferably, the collagen-based composite hemostatic powder is prepared from the following components in parts by weight: 14 parts of collagen raw material, 6 parts of modified starch, 1 part of polyethyleneimine, and 70 parts of 10-20 wt% aqueous acetic acid solution.

[0025] Preferably, in the method for preparing the modified starch, the weight ratio of starch, corn oligopeptide, and 5-15wt% boric acid aqueous solution is 7-11:0.4-1.2:30-50.

[0026] Preferably, in the method for preparing the modified starch, the weight ratio of starch, corn oligopeptide, and 5-15wt% boric acid aqueous solution is 8-10:0.6-1.0:35-45.

[0027] In one specific embodiment, the weight ratio of starch, corn oligopeptides, and 5-15 wt% boric acid aqueous solution is 7:0.4:30, 7:0.6:30, 7:0.8:30, 7:1.0:30, 7:1.2:30, 8:0.4:30, 8:0.6:30, 8:0.8:30, 8:1.0:30, 8:1.2:30, 9:0.4:30, 9:0.6:30, 9:0.8:30, 9:1.0:30, 9:1.2:30, 10:0.4:30, 10:0.6:30, 10:0.8:30, 10:1.0:30, 10:1.2:30, and 11:0. 4:30, 11:0.6:30, 11:0.8:30, 11:1.0:30, 11:1.2:30, 7:0.4:35, 7:0.6:35, 7:0.8:35, 7:1.0:35, 7:1.2:35, 8:0.4:40, 8:0.6:40, 8:0.8:40, 8:1.0:40, 8:1.2:40, 9:0.4:45, 9:0.6:45, 9:0.8:45, 9:1.0:45, 9:1.2:45, 10:0.4:50, 10:0.6:50, 10:0.8:50, 10:1.0:50, 10:1.2:50.

[0028] Experiments have shown that using the above-mentioned weight ratio of starch, corn oligopeptides, and 5-15wt% boric acid aqueous solution to prepare modified starch as a raw material component of collagen-based composite hemostatic powder can further improve the performance of collagen-based composite hemostatic powder.

[0029] Preferably, the performance parameters of the polyethyleneimine are: a 50% aqueous solution with a molecular weight of 3500-5000.

[0030] Fifthly, this application provides a method for preparing the above-mentioned collagen-based composite hemostatic powder: dissolve the collagen raw material in a 10-20 wt% aqueous acetic acid solution and mix evenly; then add modified starch at a stirring speed of 3000-6000 rpm and stir for 40-80 min; then adjust the stirring speed to 8000-12000 rpm, add polyethyleneimine, and stir for 90-150 min to obtain a slurry;

[0031] The slurry was poured into a mold and freeze-dried, and then ground to obtain the collagen-based composite hemostatic powder.

[0032] Sixthly, this application provides the application of the above-mentioned adhesive raw materials, or the above-mentioned collagen hemostatic powder, or the above-mentioned collagen-based composite hemostatic powder in the preparation of medical materials.

[0033] In summary, the technical solution of this application has the following effects:

[0034] The preparation method of the adhesive raw material provided in this application significantly reduces fat residue and greatly improves the biocompatibility of the material; it enhances the hemostatic properties of the material and promotes wound healing and tissue repair; the optimized production process makes the preparation of the material more efficient and economical, and facilitates large-scale production and clinical application.

[0035] This application utilizes a combination of collagen raw materials, modified starch, polyethyleneimine, and an aqueous acetic acid solution to prepare a collagen-based composite hemostatic powder. This powder can effectively accelerate the activation of coagulation factors, shorten coagulation time, and the hemostatic material has strong water absorption and stability. Detailed Implementation

[0036] The present application will be further described in detail below with reference to embodiments, comparative examples and performance test results. These embodiments should not be construed as limiting the scope of protection claimed in this application.

[0037] The experimental reagents used in this application were specifically sourced from the following sources: potato starch (CAS No.: 9005-25-8), purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; corn oligopeptides (item number YMDJT01) and rice oligopeptides (item number DMT01), purchased from Xi'an Shizeyuan Biotechnology Co., Ltd.; and polyethyleneimine (item number S25141, 50% aqueous solution, molecular weight 2000), polyethyleneimine (item number S25142, 50% aqueous solution, molecular weight 3500), polyethyleneimine (item number S25143, 50% aqueous solution, molecular weight 5000), and polyethyleneimine (item number S25144, 50% aqueous solution, molecular weight 10000).

[0038] Preparation Example

[0039] Preparation Examples 1-6

[0040] Preparation Examples 1-6 provide a rubber raw material and its preparation method, respectively.

[0041] The difference between the above preparation examples lies in the type of complex enzyme.

[0042] In Preparation Example 1: The complex enzyme is composed of trypsin and phospholipase in a weight ratio of 3:0.05.

[0043] In Preparation Example 2: The complex enzyme was composed of a mixture of trypsin and phospholipase in a weight ratio of 1:0.08.

[0044] In Preparation Example 3: The complex enzyme was composed of trypsin and phospholipase in a weight ratio of 5:0.02.

[0045] In Preparation Example 4: The complex enzyme was composed of a mixture of pepsin and phospholipase in a weight ratio of 3:0.05.

[0046] In Preparation Example 5: The complex enzyme was composed of a mixture of trypsin and phospholipase in a weight ratio of 0.05:3.

[0047] In Preparation Example 6: The complex enzyme was composed of a mixture of pepsin and trypsin in a weight ratio of 3:0.05.

[0048] The preparation method of the adhesive raw material in the above preparation example is as follows:

[0049] Sample solution: Take 1 kg of raw beef Achilles tendon, freeze and slice it, and place it in 10 kg of water by weight to make sample solution;

[0050] Enzymatic hydrolysis: Adjust the pH of the sample solution to 5.5, add 0.08g of compound enzyme, and enzymatically hydrolyze at 35℃ for 9h. Centrifuge and collect the supernatant. The weight ratio of compound enzyme to bovine Achilles tendon raw material is 1000:0.08. The compound enzyme is composed of trypsin and phospholipase mixed in a weight ratio of 1-5:0.02-0.08.

[0051] Alkalization reaction: Adjust the pH of the supernatant to 13.5 and alkalize it at 20℃ for 50 h;

[0052] Neutralization reaction: Adjust the pH to 7.2, elute, squeeze out water, and freeze dry to obtain the rubber raw material.

[0053] Preparation Example 7

[0054] Preparation Example 7 provides a rubber raw material and its preparation method.

[0055] The preparation method of the adhesive raw material in this preparation example is as follows:

[0056] Sample solution: Take 1 kg of raw beef Achilles tendon, freeze and slice it, and place it in 10 kg of water by weight to make sample solution;

[0057] Enzymatic hydrolysis: Adjust the pH of the sample solution to 5.5, add 0.08g of compound enzyme, and enzymatically hydrolyze at 25℃ for 9h. Centrifuge and collect the supernatant. The weight ratio of compound enzyme to bovine Achilles tendon raw material is 1000:0.08. The compound enzyme is composed of trypsin and phospholipase mixed in a weight ratio of 1-5:0.02-0.08.

[0058] Alkalization reaction: Adjust the pH of the supernatant to 13.5 and alkalize it at 20℃ for 50 h;

[0059] Neutralization reaction: Adjust the pH to 7.2, elute, squeeze out water to obtain the rubber raw material.

[0060] Preparation Example 8

[0061] Preparation Example 8 provides a rubber raw material and its preparation method.

[0062] The preparation method of the adhesive raw material in this preparation example is as follows:

[0063] Sample solution: Take 1 kg of raw beef Achilles tendon, freeze and slice it, and place it in 10 kg of water by weight to make sample solution;

[0064] Enzymatic hydrolysis: Adjust the pH of the sample solution to 5.5, add 0.08g of compound enzyme, and enzymatically hydrolyze at 35℃ for 9h. Centrifuge and collect the supernatant. The weight ratio of compound enzyme to bovine Achilles tendon raw material is 1000:0.08. The compound enzyme is composed of trypsin and phospholipase mixed in a weight ratio of 1-5:0.02-0.08.

[0065] Alkalization reaction: Adjust the pH of the supernatant to 12 and alkalize it at 20°C for 50 hours;

[0066] Neutralization reaction: Adjust the pH to 7.2, elute, squeeze out water to obtain the rubber raw material.

[0067] Preparation Example 9

[0068] Preparation Example 9 provides a rubber raw material and its preparation method.

[0069] The preparation method of the adhesive raw material in this preparation example is as follows:

[0070] Sample solution: Take 1 kg of raw beef Achilles tendon, freeze and slice it, and place it in 10 kg of water by weight to make sample solution;

[0071] Enzymatic hydrolysis: Adjust the pH of the sample solution to 5.5, add 0.08g of compound enzyme, and enzymatically hydrolyze at 35℃ for 9h. Centrifuge and collect the supernatant. The weight ratio of compound enzyme to bovine Achilles tendon raw material is 1000:0.08. The compound enzyme is composed of trypsin and phospholipase mixed in a weight ratio of 1-5:0.02-0.08.

[0072] Alkalization reaction: Adjust the pH of the supernatant to 13.5 and alkalize it at 20℃ for 50 h;

[0073] Neutralization reaction: Adjust the pH to 6.2, elute, squeeze out water to obtain the rubber raw material.

[0074] Performance testing

[0075] Collagen extraction rate test: After crushing the beef Achilles tendon raw material, it was dried in an oven at 105℃ to constant weight, and the initial dry weight was obtained. The hydroxyproline content in the beef Achilles tendon raw material and the prepared collagen raw material was tested according to GB / T9695.23-2008 "Determination of Hydroxyproline Content in Meat and Meat Products", and the collagen extraction rate in beef tendon was calculated accordingly.

[0076] Total protein content test of rubber raw materials: Bovine serum albumin was used as the standard protein, and the Folin-phenol reagent method was used for determination.

[0077] Biosafety testing of raw materials for gum: According to the cytotoxicity test method of ISO 10993-5, the effect of the extract on L929 mouse fibroblasts was evaluated by MTT method or direct contact method, and the cell survival rate was calculated.

[0078] Test results are shown in Table 1.

[0079] Table 1. Performance test results of the rubber raw materials in the preparation examples.

[0080]

[0081] As shown in the table above, Preparation Example 4 used a combination of pepsin and phospholipase at a weight ratio of 3:0.05 to prepare collagen raw materials; Preparation Example 5 used a combination of trypsin and phospholipase at a weight ratio of 0.05:3 to prepare collagen raw materials; Preparation Example 6 used a combination of pepsin and trypsin at a weight ratio of 3:0.05 to prepare collagen raw materials; Preparation Example 7 used an enzymatic hydrolysis reaction at 25°C to prepare collagen raw materials; Preparation Example 8 used an alkalization reaction at pH=12 to prepare collagen raw materials; and Preparation Example 9 used a neutralization reaction at pH=6.2 to prepare collagen raw materials. The resulting collagen raw materials had low extraction rates, low purity, and low biosafety. In contrast, the preparation examples in this application used a combination of trypsin and phospholipase at a weight ratio of 1-5:0.02-0.08 to form a complex enzyme, and then selected specific alkalization and neutralization reactions, resulting in collagen raw materials with high extraction rates, high purity, and high biosafety.

[0082] Example

[0083] Examples 1-3

[0084] Examples 1-3 provide a collagen-based composite hemostatic powder and its preparation method, respectively.

[0085] The difference in the above embodiments lies in the source of the adhesive raw materials, as detailed below.

[0086] The adhesive raw material in Example 1 was derived from Preparation Example 1.

[0087] The adhesive raw material in Example 2 was derived from Preparation Example 2.

[0088] The adhesive raw material in Example 3 was derived from Preparation Example 3.

[0089] The preparation method of the collagen-based composite hemostatic powder in the above embodiments is as follows:

[0090] (1) The modified starch is prepared by dispersing 90g starch and 8g corn oligopeptide in 400g of 10wt% boric acid aqueous solution, placing it at 50℃, mixing and stirring at 1000rpm for 75min, and then drying and grinding to obtain modified starch with an average particle size of 2-50μm.

[0091] (2) Weigh the corresponding weights of each raw material component. Take 14g of the rubber raw material (prepared in the preparation example) and dissolve it in 70g of 15wt% acetic acid aqueous solution. Mix well. Then add 6g of modified starch at a stirring speed of 4500rpm and stir for 60min. Then adjust the stirring speed to 10000rpm and add 1g of polyethyleneimine (50% aqueous solution, molecular weight 3500). Stir for 120min to obtain the slurry.

[0092] The slurry was poured into a mold and freeze-dried at -75℃ for 4 hours, dried for 25 hours, and ground until the particle size was ≤200μm to obtain collagen-based composite hemostatic powder.

[0093] Examples 4-5

[0094] Examples 4-5 respectively provide a collagen-based composite hemostatic powder and its preparation method.

[0095] The difference between the above embodiments and Embodiment 1 is that the amount of each component in the collagen-based composite hemostatic powder is different, as shown in Table 2.

[0096] Table 2. Dosage of each component in the collagen-based composite hemostatic powder in Examples 1, 4-5 and Comparative Examples 1-3

[0097]

[0098] All other process parameters in the above embodiments are the same as those in Embodiment 1.

[0099] Examples 6-12

[0100] Examples 6-12 respectively provide a collagen-based composite hemostatic powder and its preparation method.

[0101] The difference between the above embodiments and Embodiment 1 is that the preparation method of the modified starch is different, as shown below.

[0102] In Example 6: The modified starch was prepared as follows: 70g of starch and 12g of corn oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution, and the mixture was stirred at 1000rpm for 75min at 50℃. After drying and grinding, modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm were mixed in a weight ratio of 0.5:1 to obtain modified starch.

[0103] In Example 7: The modified starch was prepared as follows: 110g of starch and 4g of corn oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution. The mixture was placed at 50°C and stirred at 1000rpm for 75min. After drying and grinding, modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm were mixed in a weight ratio of 0.5:1 to obtain modified starch.

[0104] In Example 8: The modified starch was prepared as follows: 80g of starch and 10g of corn oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution, and the mixture was stirred at 1000rpm for 75min at 50℃. After drying and grinding, modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm were mixed in a weight ratio of 0.5:1 to obtain modified starch.

[0105] In Example 9: The modified starch was prepared as follows: 100g of starch and 6g of corn oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution. The mixture was placed at 50°C and stirred at 1000rpm for 75min. After drying and grinding, modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm were mixed in a weight ratio of 0.5:1 to obtain modified starch.

[0106] In Example 10: The modified starch was prepared as follows: 90g of starch and 8g of corn oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution, and the mixture was stirred at 1000rpm for 75min at 50℃. After drying and grinding, the modified starch was prepared by mixing modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm in a weight ratio of 1:0.5.

[0107] In Example 11: The modified starch was prepared as follows: 90g of starch and 8g of corn oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution, and the mixture was stirred at 1000rpm for 75min at 50℃. After drying and grinding, modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm were mixed in a weight ratio of 0.2:1 to obtain modified starch.

[0108] In Example 12: The modified starch was prepared as follows: 90g of starch and 8g of corn oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution, and the mixture was stirred at 1000rpm for 75min at 50℃. After drying and grinding, the modified starch was prepared by mixing modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm in a weight ratio of 0.6:1.

[0109] All other process parameters in the above embodiments are the same as those in Embodiment 1.

[0110] Examples 13-15

[0111] Examples 13-15 respectively provide a collagen-based composite hemostatic powder and its preparation method.

[0112] The difference between the above embodiments and Embodiment 1 is that the type of polyethyleneimine is different, as detailed below.

[0113] In Example 13, the performance parameters of polyethyleneimine were: 50% aqueous solution, molecular weight 5000.

[0114] In Example 14, the performance parameters of polyethyleneimine were: 50% aqueous solution, molecular weight 2000.

[0115] In Example 15, the performance parameters of polyethyleneimine were: 50% aqueous solution, molecular weight 10000.

[0116] All other process parameters in the above embodiments are the same as those in Embodiment 1.

[0117] Comparative Example

[0118] Comparative Examples 1-3

[0119] Comparative Examples 1-3 each provide a collagen-based composite hemostatic powder and its preparation method.

[0120] The difference between the above comparative example and Example 1 is that the amount of each component in the collagen-based composite hemostatic powder is different, as shown in Table 2.

[0121] All other process parameters in the above comparative examples are the same as those in Example 1.

[0122] Comparative Example 4-5

[0123] Comparative Examples 4 and 5 respectively provide a collagen-based composite hemostatic powder and its preparation method.

[0124] The differences between the above comparative example and Example 3 are as follows.

[0125] In Comparative Example 4, an equal amount of unmodified starch was used to replace the modified starch.

[0126] In Comparative Example 5: The modified starch was prepared as follows: 90g of starch and 8g of rice oligopeptide were dispersed in 400g of 10wt% boric acid aqueous solution and placed at 50℃. After mixing and stirring at 1000rpm for 75min, the mixture was dried and ground. Modified starch with an average particle size of 2-50μm and an average particle size of 80-200μm were mixed in a weight ratio of 0.5:1 to obtain modified starch.

[0127] All other process parameters in the above comparative examples are the same as those in Example 1.

[0128] Performance testing

[0129] (1) Hemostatic effect test: 70 SD rats weighing 280-310g were used. The temperature was 20-25℃, with sufficient light and free access to food and water. All experimental animal operation procedures complied with the "Guidelines for the Management and Use of Laboratory Animals".

[0130] Rats were allowed free access to food for 3 days before the experiment. One day before the experiment, the hair on the back was removed with 8% sodium sulfide solution. 24 hours later, the rats were disinfected with ethanol and anesthetized by intraperitoneal injection of 10% chloric acid hydrate (1-1.5 mL / kg). Using surgical scissors and forceps, two full-thickness skin layers with a diameter of 0.5 × 0.5 cm were cut off on both sides of the spine line on the back of each rat.

[0131] The animals were randomly divided into groups of 8. The collagen-based composite hemostatic powder from the examples and comparative examples was sprinkled on the wound, and gauze was laid flat on the wound. After 30 seconds, the gauze was removed to observe the bleeding. The observation was repeated every 10 seconds until the bleeding stopped. The hemostasis time was recorded and the average value was taken.

[0132] (2) Water Absorption: Weigh 1g (accurate to 0.0001g) of hemostatic powder and immerse it in ultrapure water. Every 30 minutes, remove the gelled sample, wipe off any remaining water, and weigh it. Weigh five times and record the average weight as the final mass. Place the weighed sample in a petri dish, add water again, and allow it to swell. Record the final mass as the mass after three consecutive weighings where the weight remains unchanged. The formula for calculating water absorption is: Water Absorption = (Final Mass of Sample after Water Absorption Test - Original Weight of Sample) / Original Weight of Sample × 100%

[0133] (3) Stability: The collagen-based composite hemostatic powders prepared in the examples and comparative examples were stored at room temperature in a sealed manner for 6 months and 12 months to test their hemostatic effect and water absorption, and to evaluate their stability.

[0134] Test results are shown in Table 3.

[0135] Table 3. Application performance test results of collagen-based composite hemostatic powder in the examples and comparative examples.

[0136]

[0137]

[0138] Based on the above test results, and by comparing the application performance test results of collagen-based composite hemostatic powder in the examples and comparative examples, it can be seen that the amounts of each raw material component in the collagen-based composite hemostatic powder in comparative examples 1-3 are not matched. In comparative example 4, an equal amount of unmodified starch was used to replace modified starch. In comparative example 5, modified starch was prepared by using starch, corn oligopeptides, and 5-15wt% boric acid aqueous solution. When the collagen-based composite hemostatic powder prepared was obtained, the hemostatic function and water absorption of the material were poor.

[0139] In contrast, the collagen-based composite hemostatic powder prepared using the technical solution of this application has good hemostatic function and water absorption properties, and good stability, which can effectively repair damaged and defective tissues.

[0140] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A method for preparing an adhesive raw material, characterized in that, Specifically, the following steps are performed sequentially: Sample solution preparation: Take raw bovine Achilles tendon, freeze and slice it, and place it in 8-15 times its weight of water as the sample solution; Enzymatic hydrolysis: Adjust the pH of the sample solution to 3.5-6.0, add the compound enzyme, and hydrolyze at 30-40℃ for 4-12 hours. Centrifuge and collect the supernatant. The weight ratio of the compound enzyme to the bovine Achilles tendon raw material is 0.04-0.12:1000. The compound enzyme is composed of trypsin and phospholipase mixed in a weight ratio of 1-5:0.02-0.

08. Alkalization reaction: Adjust the pH of the supernatant to >13 and alkalize it at 15-25℃ for more than 40 hours; Neutralization reaction: Adjust the pH to 6.8-7.6, elute, squeeze out water, and freeze dry to obtain the rubber raw material.

2. The method for preparing the adhesive raw material according to claim 1, characterized in that, The process parameters for the enzymatic hydrolysis reaction are: enzymatic hydrolysis at 33-38℃ for 8-10 hours.

3. The method for preparing the adhesive raw material according to claim 1, characterized in that, The weight ratio of the compound enzyme to the bovine Achilles tendon raw material is 0.06-0.10:1000; the weight ratio of the compound enzyme is 2-4: It consists of a mixture of trypsin and phospholipase at concentrations of 0.04-0.

06.

4. A collagen-based composite hemostatic powder, characterized in that, Specifically, it includes the following components: 12-16 parts of the adhesive raw material prepared by the preparation method according to any one of claims 1-3, 4-8 parts of modified starch, 0.6-1.4 parts of polyethyleneimine, and 60-80 parts of 10-20 wt% aqueous acetic acid solution; The modified starch is prepared by dispersing starch and corn oligopeptides in a 5-15 wt% boric acid aqueous solution, mixing and stirring at 40-60℃ for 60-90 minutes, and then drying and grinding to obtain modified starch.

5. The collagen-based composite hemostatic powder according to claim 4, characterized in that, In the preparation method of the modified starch, the weight ratio of starch, corn oligopeptide, and 5-15wt% boric acid aqueous solution is 7-11: 0.4-1.2：30-50。 6. The method for preparing the collagen-based composite hemostatic powder according to any one of claims 4-5, characterized in that, Dissolve the raw material in a 10-20 wt% acetic acid aqueous solution and mix well; then add the modified starch at a stirring speed of 3000-6000 rpm and stir for 40-80 min; then adjust the stirring speed to 8000-12000 rpm, add polyethyleneimine, and stir for 90-150 min to obtain a slurry. The slurry was poured into a mold and freeze-dried, and then ground to obtain the collagen-based composite hemostatic powder.

7. The application of the collagen-based composite hemostatic powder as described in claim 6 in the preparation of medical materials.

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