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A kind of production method of natural hirudin

A technology of natural hirudin and production method, applied in the direction of peptide preparation method, leech inhibitor, chemical instrument and method, etc., can solve the problems of high impurities and low purity, and achieve high product purity, high extraction rate and high quality stable effect

Active Publication Date: 2011-12-21
广西科康科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the natural hirudin that this invention extracts, its impurity is more, and purity is lower

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Take 15 kg of living Japanese leech, remove impurities, wash with water, dry naturally, put it in a refiner and refine repeatedly 3 times to obtain Japanese leech slurry; then add 150 kg of normal saline, soak for 40 minutes, and use Obtain 158.5 kilograms of filtrate after filtering through a 150-mesh sieve; add trichloroacetic acid to the above-mentioned filtrate, adjust the pH of the filtrate to 0.5 and carry out acid precipitation, leave standstill for 10 minutes, and centrifuge for 30 minutes with a 6000 rpm centrifuge to remove the precipitate, and obtain 151.3 kg of supernatant; put the above supernatant into a water bath and heat it with water. After the temperature of the supernatant in the pot reaches 85°C, continue to boil at a constant temperature for 25 minutes, take it out and cool it, and then adjust it with sodium hydroxide. pH to 7.3, then centrifuged with 10000 rev / min centrifuge for 10 minutes, to remove the precipitate, to obtain 147.6 kg of supernata...

Embodiment 2

[0041] Take 30 kg of the dead body of Hirudo phenanthae phenanthae that is frozen and preserved, put it into the refiner after it dissolves naturally, and repeatedly refine it for 4 times to obtain the Hirudo phenatus slurry; then add 60 kg of clean water to the slurry, soak for 25 minutes, After filtering with a 100-mesh sieve, 80.8 kg of filtrate was obtained; trichloroacetic acid was added to the above-mentioned filtrate, and the pH was adjusted to 3.3 for acid precipitation, left to stand for 45 minutes, centrifuged for 5 minutes with a 10000 rpm centrifuge, and the precipitate was obtained. 71.1 kg of supernatant; put the above supernatant into an electric heating jacketed pot for water-proof heating. After the temperature of the supernatant in the pot reaches 50°C, continue heating at a constant temperature for 50 minutes, take it out and cool it, and adjust it with sodium hydroxide. Adjust the pH to 6.5, then centrifuge with a centrifuge at 15,000 rpm for 15 minutes, rem...

Embodiment 3

[0043] Take 200 kg of Hirudo phenanthae live body, remove impurities, wash with water, dry naturally, put it in a refiner and repeatedly refine it twice to obtain Hirudo phenanthae slurry; then add 200 kg of clean water, soak for 30 minutes, and use After filtering through a 40-mesh sieve, 363.7 kg of filtrate was obtained; trichloroacetic acid was added to the above-mentioned filtrate, and the pH was adjusted to 1.7 for acid precipitation, left to stand for 60 minutes, centrifuged with 10000 rpm tube centrifuge, and the precipitate was obtained. 332.2 kg of supernatant; put the above supernatant into an aluminum bucket, and then boil it in a constant temperature water bath at 75°C for 20 minutes (starting from the time when the temperature of the supernatant reaches the same temperature as the water bath), Take it out for cooling, then adjust the pH to 7.0 with sodium hydroxide, then centrifuge with a tube centrifuge at 15,000 rpm to remove the precipitate, and obtain 325.3 kg...

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Abstract

The invention discloses a production method of natural hirudin. The production method is characterized by comprising the steps of extracting, acid depositing, heating, dehydrating and drying natural hirudin liquid, wherein the content of active ingredients of the obtained natural hirudin powder can be detected through Markwardt direct thrombin titration so that the quality is stable and the curative effect of the product is ensured. The method has the characteristics of simple process, high extraction ratio, low production cost, high product purity, long storage period and the like and can beused for providing safe and high-quality raw materials for foods, health-care foods, medicines or cosmetics.

Description

technical field [0001] The invention relates to a production method for extracting effective active ingredients from animal bodies, in particular to a method for producing natural hirudin by using blood-sucking leeches as raw materials. Background technique [0002] Natural hirudin is a low-molecular-weight polypeptide composed of 65-66 amino acids. Its molecular weight is only 7000 Daltons. It is easily soluble in water and saline, and it is very stable at room temperature. Simply increasing the temperature (100°C water bath), Or pH change (1 ~ 13) did not affect its activity. Natural hirudin is the strongest natural antithrombin substance found in the world so far. It has special effects on human cardiovascular and cerebrovascular diseases, especially thrombotic diseases such as cerebral thrombosis. Its preventive effect is unmatched by any substance. At the same time, relevant research also shows that natural hirudin is a direct thrombin inhibitor, its anticoagulant effe...

Claims

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Application Information

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IPC IPC(8): C07K14/815C07K1/30
Inventor 周维海周维官
Owner 广西科康科技集团有限公司
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