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Shellfish heparin with mild anticoagulant effect and its preparation method and application

An anticoagulation and shellfish technology, applied in the field of biomedicine, can solve the problems of extracting heparin, high energy consumption, and long production cycle of heparin.

Active Publication Date: 2021-04-23
GUANGDONG OCEAN UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there have been no reports on the extraction of heparin from shellfish such as sea mussels, clams, geoducks, cockles, and sand clams, let alone reports on the anticoagulant activity of these five shellfish heparins
Moreover, the extraction and production cycle of existing heparin is long, the energy consumption is high, and the activity loss of the product is large.

Method used

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  • Shellfish heparin with mild anticoagulant effect and its preparation method and application
  • Shellfish heparin with mild anticoagulant effect and its preparation method and application
  • Shellfish heparin with mild anticoagulant effect and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The preparation of embodiment 1 shellfish heparin

[0052] 1. A method for preparing shellfish heparin, comprising the following steps:

[0053] S1. Raw material treatment: wash shellfish raw materials, after shelling, put shellfish meat in acetone at 2°C and soak for 20 hours, remove acetone by suction filtration at room temperature, rinse with distilled water for 3 to 5 times, then add distilled water (solid-liquid ratio =1:3) for homogenization to obtain a homogenate solution; wherein, the shellfish is selected from any one of sea mussels, geoducks, mud clams, clams or clams.

[0054] S2. Autolysis: autolyze the homogenate in a water bath at 50°C for 5 hours;

[0055] S3. Enzymolysis: add NaOH to adjust the pH to 8.0, first add 0.5% trypsin, and enzymolyze at 37°C for 5 hours, then add HCl to adjust the pH to 7.0, then add 0.5% papain, and enzymolyze at 65°C 5h, to obtain the enzymatic solution;

[0056] S4. Enzyme inactivation and centrifugation: put the enzymati...

Embodiment 2

[0061] The preparation of embodiment 2 shellfish heparin

[0062] 1. A method for preparing shellfish heparin, comprising the following steps:

[0063] S1. Raw material processing: wash the shellfish raw material, remove the shell, add distilled water (material-to-liquid ratio=1:1.5) for homogenization, and obtain a homogenate;

[0064] S2. Autolysis: autolyze the homogenate in a water bath at 38°C for 7 hours;

[0065] S3. Enzymolysis: add NaOH to adjust the pH to 7.8, first add 0.3% trypsin, and enzymatically hydrolyze at 32°C for 6.5h, then add HCl to adjust the pH to 6.8, then add 0.3% papain, and enzymatically digest at 62°C Decompose for 5.5h to obtain the enzymatic solution;

[0066] S4. Enzyme inactivation and centrifugation: put the enzymatic solution in boiling water to inactivate the enzyme for 5 minutes, and centrifuge at 8000 rpm for 10 minutes;

[0067] S5. Concentration and alcohol precipitation: After concentrating the supernatant, add 0.3 times the volume o...

Embodiment 3

[0071] The preparation of embodiment 3 shellfish heparin

[0072] 1. A method for preparing shellfish heparin, comprising the following steps:

[0073] S1. Raw material treatment: wash shellfish raw materials, after shelling, put shellfish meat in acetone at 6°C and soak for 28 hours, remove acetone by suction filtration at room temperature, rinse with distilled water for 3 to 5 times, then add distilled water (ratio of solid to liquid) =1:4.5) for homogenization to obtain a homogenate;

[0074] S2. Autolysis: autolyze the homogenate in a water bath at 62°C for 3 hours;

[0075] S3. Enzymolysis: add NaOH to adjust the pH to 8.2, first add 0.7% trypsin, enzymatically hydrolyze at 42°C for 3.5h, then add HCl to adjust the pH to 7.2, then add 0.7% papain, enzymatically at 68°C Decompose for 4.5h to obtain the enzymatic solution;

[0076] S4. Enzyme inactivation and centrifugation: put the enzymatic solution in boiling water to inactivate the enzyme for 15 minutes, and centrifu...

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Abstract

The invention discloses shellfish heparin with mild anticoagulant effect, its preparation method and application. The preparation method is: raw material treatment; autolysis; enzymatic hydrolysis: adding NaOH to adjust the pH to 7.8-8.2, adding trypsin, enzymatic hydrolysis at 32-42°C for 3.5-6.5 h, then adding HCl to adjust the pH to 6.8-7.2 , add papain, and enzymolyze at 62-68°C for 4.5-5.5 h to obtain an enzymatic solution; inactivate enzyme, centrifuge; concentrate, alcohol precipitation; redissolve, deproteinize; desalt; dry. The present invention is the first to prepare shellfish heparin with anticoagulant effect from sea mussels, geoducks, clams, cockles, and scallops. The heparin comes from marine shellfish, has high safety and moderate action. The invention not only widens the source of heparin, but also has simple preparation method and stable product quality, and has good application value in the preparation of heparin with low side effect or other products with mild anticoagulant effect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. More specifically, it relates to a shellfish heparin with mild anticoagulant effect, its preparation method and application. Background technique [0002] Heparin is a glycosaminoglycan that exists in animals. It has various biological activities and is widely used clinically, especially as an anticoagulant. At present, there is no product that can completely replace it. Most of the heparin used clinically comes from porcine small intestine and bovine lung, but the heparin derived from porcine small intestine and bovine lung has too strong anticoagulant effect, which is likely to cause side effects such as bleeding effect and decreased platelets. In view of the problems of excessive anticoagulant activity and too many side effects of porcine and bovine heparin, it is necessary to develop heparin with moderate anticoagulant effect. Marine heparin has the characteristics of good compatibility...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/10A61P7/02
CPCA61P7/02C08B37/0003C08B37/0075
Inventor 钟赛意杜振兴周斯仪陈观兰陈菁陈建平谌素华洪鹏志
Owner GUANGDONG OCEAN UNIVERSITY
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