Compound frequently combined ultrasonic reinforced double-enzyme heparin sodium extracting technique

A technology of heparin sodium and ultrasound, applied in the field of heparin sodium, can solve the problems of reduced yield of heparin sodium, difficult to achieve titer, and great influence on the biological activity of heparin sodium.

Inactive Publication Date: 2009-01-14
罗时通
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the technological process of producing heparin sodium by the above-mentioned method is relatively simple and easy to realize, the weak point of existence is that the process is delicate, because the crude product heparin sodium is thro...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Make small intestine mucosa (10 pairs of small intestines), about 4-6 kg after each small intestine is mixed with water, pump the small intestines scraped into a conventional enzymolysis tank, add 2.3-2.8% (volume) of Refined salt, adjust the pH value to 9.0-9.2; add trypsin and lipase (ratio: 10:2) at 42°C, accounting for 0.4% by weight of the total solution; continue to heat up to 52°C, keep warm for 3 hours and then carry out enzymatic hydrolysis , then continue to heat up to 80°C-85°C, keep warm for 15 minutes and then filter, then adsorb the filtered solution in the above steps: filter with adsorption water, add resin to absorb when cooled to 58°C; wash: collect resin, rinse with water several times , and then heated with water to 58-60°C, adjusted the pH to 7.5, until the resin was washed clean, used 1.2N saline solution, adjusted the pH to 8, added to the resin, heated to 60°C, washed for 0.5h, removed the washing solution, Filter and dry the resin; elution: afte...

Embodiment 2

[0021] Scrape the small intestine mucosa (10 pairs of small intestines), mix each small intestine with water about 4-6 kg, pump the scraped small intestine into the multiple frequency ultrasonic reactor (20+40kHZ), add 2.3- The refined salt of 2.8% (volume) adjusts pH value 9.0-9.2; Add trypsin and lipase (ratio is 10: 2) when 42 ℃, account for total solution weight ratio and be 0.4%; Continue heating to 52 ℃, After 3 hours of heat preservation, carry out enzymatic hydrolysis, then continue to heat up to 80°C-85°C, heat for 15 minutes, then filter, and carry out the normal steps of adsorption, washing, elution, precipitation, dehydration and drying of the filtered solution.

[0022] The yield comparison table for products with different ultrasonic intensities:

[0023] Ultrasound intensity

[0024] It can be seen from the above table that the optimal ultrasonic intensity is 0.5W / m 2 .

Embodiment 3

[0026] Scrape the small intestine mucosa (10 pairs of small intestines), mix each small intestine with water about 4-6 kg, pump the scraped small intestine into a multiple frequency ultrasonic reactor (20+40kHZ), and the ultrasonic intensity is 0.5W / m 2 , before heating, add 2.3-2.8% refined salt to adjust the pH value to 9.0-9.2; add trypsin and lipase (ratio is 10: 2) at 42°C, accounting for 0.4% by weight of the total solution; continue to add Warm to 52°C, keep warm for different times, then carry out enzymatic hydrolysis, then continue to heat up to 80°C-85°C, keep warm for about 15 minutes, then filter, and carry out the normal steps of adsorption, washing, elution, precipitation, dehydration and drying of the filtered solution.

[0027] Product yield comparison table under different ultrasonic enzyme time:

[0028] During sonication

[0029] It can be seen from this that only 2 hours of enzymatic hydrolysis is required, which saves about 33% in time compared w...

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PUM

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Abstract

The invention relates to a heparin sodium technique, in particular to a heparin sodium extracting technique utilizing multi-frequency combination and ultrasonic double-enzyme reinforcement. Firstly, the scraped small intestinal mucosa is sucked in a conventional enzymolysis vessel, and refined salt with a proportion of 2.3 percent to 2.8 percent by weight is firstly added thereto before heating, and the pH value is adjusted to 9.0 to 9.2; secondly, when the solution obtained from the first step is heated to 40 DEG C to 45 DEG C, trypsase and lipase are added, and the proportion is maintained between 10 : 0.5 to 10 : 5, the weight proportion of the double enzyme in the solution in the first step accounts for 0.3 percent to 0.4 percent, the solution is continuously heated to 50 DEG C to 55 DEG C, the enzymolysis under heat preservation is performed for 2 hours to 5 hours, then the solution is continuously heated to 80 DEG C to 85 DEG C, and the solution is filtered after being insulated for 10 minutes to 20 minutes; and thirdly, the filtered solution obtained from the second step is treated through the conventional steps of adsorbing, washing, eluting, precipitating, dehydrating and drying, so as to obtain the heparin sodium product. Compared with the prior art, the invention has the advantages that: firstly, the entire procedure time is substantially reduced by 33 percent; secondly, the yield of products is increased by 29.6 percent; and thirdly, the titer is stabilized to be higher than 120 (the titer of the domestic advanced technology product is between 90 and 110).

Description

Contents of the invention [0001] The present invention relates to a technology of heparin sodium, in particular to the background technology of the technology of double-enzyme extraction of heparin sodium combined with multiple frequency combination ultrasound. Background technique [0002] At present, almost all production in our country uses pig small intestine as raw material, and several methods such as salt hydrolysis, alkali hydrolysis and enzymatic hydrolysis are used in conventional reactors, among which salt hydrolysis is the most common application, and its production process is: raw material (intestinal mucosa), conventional reactor Inner brine extraction, filtration, resin adsorption, elution, ethanol precipitation, dehydration, drying, crude heparin sodium. [0003] Although the process of producing heparin sodium by the above method is relatively simple and easy to realize, the weak point of existence is that the process is delicate, because the crude product h...

Claims

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Application Information

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IPC IPC(8): C12P19/04
Inventor 罗时通
Owner 罗时通
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