Novel environment-friendly biological liquid bone glue preparation process

Abstract

The invention provides a novel environment-friendly biological liquid bone glue preparation process, and relates to the technical field of biological environment-friendly material preparation. The preparation process of the novel environment-friendly biological liquid bone glue comprises the following steps: preparing the following raw materials: 90g of bone glue, 100g of water, a proper amount of protease, 8g of acrylic acid, 7.6 g of ammonium thiocyanate, 10ml of methanol and 10ml of coral dealbatus extracting solution; decomposing the bone glue, namely adding 90g of bone glue into a 500ml volumetric flask, adding 100g of water, uniformly stirring to fully dissolve the bone glue, adding a proper amount of protease, carrying out enzymolysis reaction at normal temperature, and uniformly stirring; heating in a water bath: putting the bone glue solution subjected to enzymolysis into a water bath kettle, heating to 80 DEG C, keeping stirring in the heating process to ensure that the temperature is uniform, and keeping the temperature for 10 minutes after heating to 80 DEG C to promote the complete proceeding of the enzymolysis reaction. Unsaturated monomers such as acrylic acid are introduced for chemical crosslinking, so that the mechanical strength and stability of the bone glue are remarkably improved.

Description

Technical Field

[0001] The present invention relates to the technical field of preparation of environmentally friendly biological materials, in particular to a novel process for preparing environmentally friendly biological liquid bone glue. Background Art

[0002] As global environmental awareness continues to increase and the concept of sustainable development becomes more popular, the demand for green and environmentally friendly materials in all walks of life is increasing. In the field of materials science, traditional bone glue has been widely used in food, medicine, cosmetics, adhesives and other industries due to its excellent bonding properties and biodegradability.

[0003] In recent years, with the advancement of biotechnology, the application of enzymatic hydrolysis technology in material preparation has gradually attracted attention. Enzymatic hydrolysis technology uses specific biological enzymes to selectively decompose raw materials, and can achieve the preparation of target products under relatively mild conditions. Compared with traditional high-temperature and high-pressure treatment, enzymatic hydrolysis technology has the advantages of low energy consumption, mild reaction conditions, and environmental friendliness. Through enzymatic hydrolysis technology, the molecular weight and properties of bone glue can be more accurately controlled, thereby preparing bone glue products with better performance. However, traditional bone glue preparation processes usually rely on high-temperature and high-pressure treatment. This method not only has high energy consumption, but also has a certain impact on the environment. In addition, traditional bone glue may have problems such as insufficient stability and poor anti-corrosion performance in certain application scenarios, which limits its wider application. Therefore, technical personnel in this field provide a new environmentally friendly biological liquid bone glue preparation process to solve the problems raised in the above background technology. Summary of the Invention

[0004] (1) Technical problems solved In view of the shortcomings of the existing technology, the present invention provides a new environmentally friendly biological liquid bone glue preparation process, which solves the problems of high energy consumption, insufficient stability and poor anti-corrosion performance in the existing preparation process.

[0005] (2) Technical solution To achieve the above objectives, the present invention is implemented through the following technical solutions: A novel environmentally friendly biological liquid bone glue preparation process, comprising: Prepare the raw materials, including 90g of bone glue, 100g of water, an appropriate amount of protease, 8g of acrylic acid, 7.6g of ammonium thiocyanate, 10ml of methanol and 10ml of schizonepeta tenuifolia extract; For gelatin decomposition, add 90g of gelatin into a 500ml volumetric flask, add 100g of water, stir evenly to fully dissolve the gelatin, add appropriate amount of protease, carry out enzymatic hydrolysis at room temperature, and stir evenly; Heat the enzymatically hydrolyzed gelatin solution in a water bath and heat it to 80°C. Stir the solution during heating to ensure uniform temperature. After heating to 80°C, maintain this temperature for 10 minutes to promote the complete enzymatic hydrolysis reaction. Chemical crosslinking: add 8g of acrylic acid to the gelatin solution at 80°C and stir evenly to allow the acrylic acid to fully react with the gelatin to form a crosslinked structure. The reaction time is 30 to 60 minutes. Add 7.6g of ammonium thiocyanate as a fixative to the above solution and stir evenly to allow the ammonium thiocyanate to react fully. The reaction time is 15 to 30 minutes; Add anticoagulant and preservative: add 10 ml of methanol as anticoagulant to the reaction system, stir evenly, then add 10 ml of radix serrata extract as preservative, stir evenly; After the reaction is completed, continue stirring the reaction system for 10 minutes to ensure that all components are fully mixed and reacted. After the reaction is completed, stop heating and let it cool naturally to room temperature to obtain a new environmentally friendly biological liquid bone glue.

[0006] Preferably, the decomposition of bone gelatin comprises the following steps: S1. Weighing and dissolving bone glue: Use a high-precision electronic balance to accurately weigh 90 g of bone glue; S2. Place the weighed amount of bone glue into a clean 500 mL volumetric flask. S3. Use a graduated cylinder to measure 100 mL of distilled or deionized water and slowly pour it into the volumetric flask. S4. Use a glass rod to gently stir to initially disperse the gelatin in the water. S5. Determine the amount of protease to be used based on its activity. Slowly add the protease to the volumetric flask and gently stir with a glass rod to evenly distribute the protease in the gelatin solution. S6. Place the volumetric flask on a thermostatic magnetic stirrer, set the temperature to room temperature, turn on the stirrer, adjust the speed to 200 rpm to 300 rpm, and proceed with the enzymatic hydrolysis reaction for 2 to 4 hours, the specific time being determined by the characteristics of the gelatin and the activity of the protease.

[0007] Preferably, an appropriate amount of catalyst may be added during the chemical cross-linking process, and the catalyst is ammonium sulfate or methylenebisacrylamide.

[0008] Preferably, the protease used in the decomposition of bone gelatin is alkaline protease or neutral protease, the amount of enzyme added is 0.48 g, and the pH value is adjusted to 8.5±0.1 using sodium bicarbonate solution.

[0009] Preferably, after the reaction is completed, the viscosity of the product is measured using a viscometer and compared with the standard value to ensure that the product meets expectations. The product is placed at different temperatures to observe its stability, and an accelerated aging test is performed to evaluate the long-term stability of the product. At the same time, a microbial challenge test is performed to evaluate the antiseptic effect of the product, and the microbial content of the product is regularly tested to ensure its safety.

[0010] Preferably, the chemical crosslinking comprises the following steps: S1. Preparation of a crosslinker: using acrylic acid monomer, purified by vacuum distillation or using an inhibitor scavenger to ensure its reactivity, dissolving the purified acrylic acid monomer in an appropriate amount of deionized water to prepare a solution of a certain concentration; S2. Implementation of the cross-linking reaction: Transfer the enzymatically hydrolyzed gelatin solution to a clean container and adjust the pH of the solution to 7.0 to 8.0 to ensure optimal reactivity of acrylic acid; S3. Addition of an initiator: adding an appropriate amount of initiator to the gelatin solution in an amount of 0.1% to 1.0% by mass of acrylic acid; S4. The cross-linking agent is added, and the prepared acrylic acid solution is slowly added to the gelatin solution while stirring, maintaining a stirring speed of 300rpm to 500rpm to ensure that the acrylic acid is evenly distributed in the gelatin solution; S5. Control the reaction conditions, heat the reaction system to 80°C to 90°C to promote the decomposition of the initiator and the generation of free radicals, maintain the temperature for 30 to 60 minutes, and after the reaction is completed, stop heating and naturally cool to room temperature.

[0011] Preferably, the adding of the anticoagulant and preservative comprises the following steps: S1. Methanol was added. 10 ml of the prepared methanol was slowly added to the reaction system. While adding methanol, stirring was continued to ensure that the methanol was evenly distributed in the reaction system. After methanol was added, the reaction system was stirred for 10 minutes to ensure that the methanol and the gelatin were fully mixed. S2. Add preservatives, select fresh Chinese herb, rinse it with clean water to remove dirt and impurities, then dry it in a cool place, crush the dried Chinese herb into coarse powder, use water to perform decoction extraction, after the extraction is completed, filter to remove solid residue to obtain Chinese herb extract, concentrate the extract to obtain a concentrated solution, dissolve the Chinese herb extract or extract in an appropriate amount of deionized water to prepare a solution of a certain concentration, and slowly add the prepared Chinese herb extract to the reaction system.

[0012] (3) Beneficial effects The present invention provides a novel environmentally friendly biological liquid bone glue preparation process, which has the following beneficial effects: In the present invention, enzymatic hydrolysis technology is adopted to carry out the reaction at room temperature or relatively low temperature, thereby avoiding the high energy consumption problem caused by high temperature and high pressure treatment in the traditional bone glue preparation process. Less waste water and waste gas are generated during the reaction process, and the impact on the environment is relatively small. The prepared bone glue product itself has good biodegradability and will not cause long-term burden on the environment after use, which is in line with the concept of sustainable development.

[0013] In the present invention, the enzymatic hydrolysis technology can achieve sufficient decomposition of bone gelatin in a relatively short time, significantly shortening the production cycle and improving production efficiency.

[0014] In the present invention, by introducing unsaturated monomers such as acrylic acid for chemical cross-linking, the mechanical strength and stability of bone glue are significantly improved, so that it can maintain good performance in various application scenarios. By adjusting the enzymatic hydrolysis conditions, the types and amounts of chemical cross-linking agents and preservatives, bone glue products with different performance characteristics can be prepared to meet the needs of different application scenarios.

[0015] In the present invention, by using natural plant extracts as preservatives, potential safety hazards and environmental pollution problems caused by chemical preservatives are avoided. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Schematic diagram of the overall system process of the present invention. DETAILED DESCRIPTION

[0017] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. 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 making creative efforts are within the scope of protection of the present invention.

[0018] Example 1 like Figure 1 As shown, the embodiment of the present invention provides a novel environmentally friendly biological liquid bone glue preparation process, comprising: Prepare the raw materials, including 90g of bone glue, 100g of water, an appropriate amount of protease, 8g of acrylic acid, 7.6g of ammonium thiocyanate, 10ml of methanol and 10ml of schizonepeta tenuifolia extract; For gelatin decomposition, add 90g of gelatin into a 500ml volumetric flask, add 100g of water, stir evenly to fully dissolve the gelatin, add appropriate amount of protease, carry out enzymatic hydrolysis at room temperature, and stir evenly; Heat the enzymatically hydrolyzed gelatin solution in a water bath and heat it to 80°C. Stir the solution during heating to ensure uniform temperature. After heating to 80°C, maintain this temperature for 10 minutes to promote the complete enzymatic hydrolysis reaction. Chemical crosslinking: add 8g of acrylic acid to the gelatin solution at 80°C and stir evenly to allow the acrylic acid to fully react with the gelatin to form a crosslinked structure. The reaction time is 30 to 60 minutes. Add 7.6g of ammonium thiocyanate as a fixative to the above solution and stir evenly to allow the ammonium thiocyanate to react fully. The reaction time is 15 to 30 minutes; Add anticoagulant and preservative: add 10 ml of methanol as anticoagulant to the reaction system, stir evenly, then add 10 ml of radix serrata extract as preservative, stir evenly; After the reaction is completed, continue stirring the reaction system for 10 minutes to ensure that all components are fully mixed and reacted. After the reaction is completed, stop heating and let it cool naturally to room temperature to obtain a new environmentally friendly biological liquid bone glue.

[0019] The decomposition of bone gelatin involves the following steps: S1. Weighing and dissolving bone glue: Use a high-precision electronic balance to accurately weigh 90 g of bone glue; S2. Place the weighed amount of bone glue into a clean 500 mL volumetric flask. S3. Use a graduated cylinder to measure 100 mL of distilled or deionized water and slowly pour it into the volumetric flask. S4. Use a glass rod to gently stir to initially disperse the gelatin in the water. S5. Determine the amount of protease to be used based on its activity. Slowly add the protease to the volumetric flask and gently stir with a glass rod to evenly distribute the protease in the gelatin solution. S6. Place the volumetric flask on a thermostatic magnetic stirrer, set the temperature to room temperature, turn on the stirrer, adjust the speed to 200 rpm to 300 rpm, and proceed with the enzymatic hydrolysis reaction for 2 to 4 hours, the specific time being determined by the characteristics of the gelatin and the activity of the protease.

[0020] During the chemical cross-linking process, an appropriate amount of catalyst can be added, and the catalyst is ammonium sulfate or methylene bisacrylamide.

[0021] The protease used in gelatin decomposition is alkaline protease or neutral protease, the amount of enzyme added is 0.48 g, and the pH value is adjusted to 8.5±0.1 using sodium bicarbonate solution.

[0022] After the reaction is completed, the viscosity of the product is measured using a viscometer and compared with the standard value to ensure that the product meets expectations. The product is placed at different temperatures to observe its stability and conduct accelerated aging tests to evaluate the long-term stability of the product. At the same time, microbial challenge tests are conducted to evaluate the antiseptic effect of the product. The microbial content of the product is regularly tested to ensure its safety.

[0023] Chemical cross-linking involves the following steps: S1. Preparation of a crosslinker: using acrylic acid monomer, purified by vacuum distillation or using an inhibitor scavenger to ensure its reactivity, dissolving the purified acrylic acid monomer in an appropriate amount of deionized water to prepare a solution of a certain concentration; S2. Implementation of the cross-linking reaction: Transfer the enzymatically hydrolyzed gelatin solution to a clean container and adjust the pH of the solution to 7.0 to 8.0 to ensure optimal reactivity of acrylic acid; S3. Addition of an initiator: adding an appropriate amount of initiator to the gelatin solution in an amount of 0.1% to 1.0% by mass of acrylic acid; S4. The cross-linking agent is added, and the prepared acrylic acid solution is slowly added to the gelatin solution while stirring, maintaining a stirring speed of 300rpm to 500rpm to ensure that the acrylic acid is evenly distributed in the gelatin solution; S5. Control the reaction conditions, heat the reaction system to 80°C to 90°C to promote the decomposition of the initiator and the generation of free radicals, maintain the temperature for 30 to 60 minutes, and after the reaction is completed, stop heating and naturally cool to room temperature.

[0024] The addition of anticoagulant and preservative comprises the following steps: S1. Methanol was added. 10 ml of the prepared methanol was slowly added to the reaction system. While adding methanol, stirring was continued to ensure that the methanol was evenly distributed in the reaction system. After methanol was added, the reaction system was stirred for 10 minutes to ensure that the methanol and the gelatin were fully mixed. S2. Add preservatives, select fresh Chinese herb, rinse it with clean water to remove dirt and impurities, then dry it in a cool place, crush the dried Chinese herb into coarse powder, use water to perform decoction extraction, after the extraction is completed, filter to remove solid residue to obtain Chinese herb extract, concentrate the extract to obtain a concentrated solution, dissolve the Chinese herb extract or extract in an appropriate amount of deionized water to prepare a solution of a certain concentration, and slowly add the prepared Chinese herb extract to the reaction system.

[0025] While 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 these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A novel environmentally friendly biological liquid bone glue preparation process, characterized by: include: Prepare the raw materials, including 90g of bone glue, 100g of water, an appropriate amount of protease, 8g of acrylic acid, 7.6g of ammonium thiocyanate, 10ml of methanol and 10ml of schizonepeta tenuifolia extract; For gelatin decomposition, add 90g of gelatin into a 500ml volumetric flask, add 100g of water, stir evenly to fully dissolve the gelatin, add appropriate amount of protease, carry out enzymatic hydrolysis at room temperature, and stir evenly; Heat the enzymatically hydrolyzed gelatin solution in a water bath and heat it to 80°C. Stir the solution during heating to ensure uniform temperature. After heating to 80°C, maintain this temperature for 10 minutes to promote the complete enzymatic hydrolysis reaction. Chemical crosslinking: add 8g of acrylic acid to the gelatin solution at 80°C and stir evenly to allow the acrylic acid to fully react with the gelatin to form a crosslinked structure. The reaction time is 30 to 60 minutes. Add 7.6g of ammonium thiocyanate as a fixative to the above solution and stir evenly to allow the ammonium thiocyanate to react fully. The reaction time is 15 to 30 minutes; Add anticoagulant and preservative: add 10 ml of methanol as anticoagulant to the reaction system, stir evenly, then add 10 ml of radix serrata extract as preservative, stir evenly; After the reaction is completed, continue stirring the reaction system for 10 minutes to ensure that all components are fully mixed and reacted. After the reaction is completed, stop heating and let it cool naturally to room temperature to obtain a new environmentally friendly biological liquid bone glue.

2. The novel environmentally friendly biological liquid bone glue preparation process according to claim 1, characterized in that: The decomposition of bone gelatin comprises the following steps: S1. Weighing and dissolving bone glue: Use a high-precision electronic balance to accurately weigh 90 g of bone glue; S2. Place the weighed amount of bone glue into a clean 500 mL volumetric flask. S3. Use a graduated cylinder to measure 100 mL of distilled or deionized water and slowly pour it into the volumetric flask. S4. Use a glass rod to gently stir to initially disperse the gelatin in the water. S5. Determine the amount of protease to be used based on its activity. Slowly add the protease to the volumetric flask and gently stir with a glass rod to evenly distribute the protease in the gelatin solution. S6. Place the volumetric flask on a thermostatic magnetic stirrer, set the temperature to room temperature, turn on the stirrer, adjust the speed to 200 rpm to 300 rpm, and proceed with the enzymatic hydrolysis reaction for 2 to 4 hours, the specific time being determined by the characteristics of the gelatin and the activity of the protease.

3. The novel environmentally friendly biological liquid bone glue preparation process according to claim 1, characterized in that: During the chemical cross-linking process, a suitable amount of catalyst may be added, and the catalyst is ammonium sulfate or methylene bisacrylamide.

4. The novel environmentally friendly biological liquid bone glue preparation process according to claim 1, characterized in that: The protease used in the gelatin decomposition is alkaline protease or neutral protease, the amount of enzyme added is 0.48 g, and the pH value is adjusted to 8.5±0.1 using sodium bicarbonate solution.

5. The novel environmentally friendly biological liquid bone glue preparation process according to claim 1, characterized in that: After the reaction is completed, the viscosity of the product is measured using a viscometer and compared with the standard value to ensure that the product meets expectations. The product is placed at different temperatures to observe its stability, and an accelerated aging test is performed to evaluate the long-term stability of the product. At the same time, a microbial challenge test is performed to evaluate the antiseptic effect of the product, and the microbial content of the product is regularly tested to ensure its safety.

6. The novel environmentally friendly biological liquid bone glue preparation process according to claim 1, characterized in that: The chemical cross-linking comprises the following steps: S1. Preparation of a crosslinker: using acrylic acid monomer, purified by vacuum distillation or using an inhibitor scavenger to ensure its reactivity, dissolving the purified acrylic acid monomer in an appropriate amount of deionized water to prepare a solution of a certain concentration; S2. Implementation of the cross-linking reaction: Transfer the enzymatically hydrolyzed gelatin solution to a clean container and adjust the pH of the solution to 7.0 to 8.0 to ensure optimal reactivity of acrylic acid; S3. Addition of an initiator: adding an appropriate amount of initiator to the gelatin solution in an amount of 0.1% to 1.0% by mass of acrylic acid; S4. The cross-linking agent is added, and the prepared acrylic acid solution is slowly added to the gelatin solution while stirring, maintaining a stirring speed of 300rpm to 500rpm to ensure that the acrylic acid is evenly distributed in the gelatin solution; S5. Control the reaction conditions, heat the reaction system to 80°C to 90°C to promote the decomposition of the initiator and the generation of free radicals, maintain the temperature for 30 to 60 minutes, and after the reaction is completed, stop heating and naturally cool to room temperature.

7. The novel environmentally friendly biological liquid bone glue preparation process according to claim 1, characterized in that: The addition of the anticoagulant and preservative comprises the following steps: S1. Methanol was added. 10 ml of the prepared methanol was slowly added to the reaction system. While adding methanol, stirring was continued to ensure that the methanol was evenly distributed in the reaction system. After methanol was added, the reaction system was stirred for 10 minutes to ensure that the methanol and the gelatin were fully mixed. S2. Add preservatives, select fresh Chinese herb, rinse it with clean water to remove dirt and impurities, then dry it in a cool place, crush the dried Chinese herb into coarse powder, use water to perform decoction extraction, after the extraction is completed, filter to remove solid residue to obtain Chinese herb extract, concentrate the extract to obtain a concentrated solution, dissolve the Chinese herb extract or extract in an appropriate amount of deionized water to prepare a solution of a certain concentration, and slowly add the prepared Chinese herb extract to the reaction system.

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