阅读说明：本技术 盐解法利用母猪肠制备粗品肝素钠的方法 (Method for preparing crude heparin sodium by using sow intestines through salt hydrolysis method ) 是由 辛玉昌 黄蓉屏 范时兵 于 2019-09-27 设计创作，主要内容包括：本发明公开了盐解法利用母猪肠制备粗品肝素钠的方法。本发明中包括以下步骤：肠粘膜提取、盐解、分离、降温、吸附、树脂收集以及固化,本发明能够增加油脂与蛋白质之间的分离效果,降低生产成本,技术成熟、实用,肝素收率高,工艺品质稳定。(The invention discloses a method for preparing crude heparin sodium by utilizing sow intestines through a salt hydrolysis method. The invention comprises the following steps: the invention can increase the separation effect between the grease and the protein, reduce the production cost, has mature and practical technology, high heparin yield and stable process quality.)

1. The salt hydrolysis method for preparing crude heparin sodium by utilizing sow intestines comprises the following steps:

(1) intestinal mucosa extraction

Rolling healthy fresh sow intestines with deionized water by using a rolling machine for multiple times, then feeding the rolled intestines into an intestine scraping machine for scraping, and collecting intestinal mucosa mixed liquor prepared in the rolling and scraping processes;

(2) salt decomposition

Pumping the intestinal mucosa mixed solution into a salt decomposition tank by using a lift pump, stirring at normal temperature, adjusting the salinity of the mixed solution to 4.2 +/-0.3 ℃ by using saturated concentrated brine, adjusting the pH to 9.0 by using a sodium hydroxide solution, continuously heating to 58 +/-1 ℃, then adding trypsin, and carrying out heat preservation and deproteinization to ensure that the protein in the salt decomposition solution floats on the liquid surface after being solidified;

(3) separation of

Respectively separating oil and protein from the salinized solution through an intestinal residue filter barrel and a high-speed liquid-solid three-phase separator to obtain the salinized solution after protein and oil are separated, and pouring the salinized solution into a buffer tank;

(4) temperature reduction

Cooling the salt solution by using a plate heat exchanger to obtain an adsorption solution;

(5) adsorption

The adsorption solution is pumped into an adsorption tank, the pH value is adjusted to be more than 8.0, the salinity is more than 3.0, the treated resin is added for ion adsorption, and the adsorption is accelerated by stirring;

(6) resin collection

Collecting the adsorbed resin by a resin collecting tank, and pumping the resin into a rinsing tank for thorough cleaning to remove impurities such as protein, grease and the like;

(7) curing

And eluting, precipitating and drying the collected resin by using a traditional process to obtain a crude product of heparin sodium.

2. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the water quantity of the deionized water used in the step (1) is controlled to be 5 +/-1 Kg per sow intestine.

3. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the stirring speed used in step (2) was 60. + -. 5 rpm.

4. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the concentration of the sodium hydroxide solution used in the steps (2) and (5) is 0.3 +/-0.2 mol/L.

5. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the method comprises the following steps:

preparing an enzyme inactivation solution;

(1) one-time heat preservation

Keeping the temperature of the mixture at 58 + -1 deg.C, pH 8.5, salinity > 3.1, and maintaining for 120 min;

(2) high temperature inactivation

Rapidly heating to 96 + -1 deg.C, adjusting pH to 8.3 + -1, and maintaining for 10min to make protein in the salt solution float on the liquid surface after fixing.

6. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the method comprises the following steps:

preparing a salinization solution;

(1) rough fraction

Performing solid-liquid separation on the salinization liquid by using an intestinal residue filtering roller, and enabling the separated salinization liquid to enter a buffer tank;

(2) finely dividing

And (3) performing oil and protein fine separation on the coarsely separated salinized solution again by using a high-speed liquid-solid three-phase separator with the separation factor of 12200 +/-200 g and the rpm of 6400 +/-300 to obtain a second salinized solution.

7. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the temperature of the step (4) is controlled to be 60 +/-2 ℃.

8. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the resin in the step (5) is Bayer resin, and the dosage of the resin is 1Kg of Bayer resin added to every 60Kg of small intestine of a pig.

9. The salting process of claim 1 for preparing crude heparin sodium from the intestines of sows, comprising the steps of: the stirring speed in the step (5) is 55 +/-5 rpm, and the stirring time is 420 +/-60 min.

Technical Field

The invention belongs to a preparation method of crude heparin sodium, and particularly relates to a method for preparing crude heparin sodium by utilizing sow intestines through a salt hydrolysis method.

Background

Heparin sodium interferes with many links of the blood coagulation process, has an anticoagulant effect both in vitro and in vivo, has a complex action mechanism, enhances the inhibition effect of the antithrombin III on activated blood coagulation factors II, IX, X, XI and XII mainly through the combination with the antithrombin III, and has the effects of preventing platelet aggregation and destruction, preventing the formation of blood coagulation activating enzyme, preventing prothrombin from being changed into thrombin, inhibiting thrombin, preventing fibrinogen from being changed into fibrin and exerting the anticoagulant effect.

The technical problems in production of extracting heparin sodium in a sow farm mainly include high production cost, low heparin yield and lack of competitive advantages due to the fact that the oil and protein separation technology is not solved for a long time.

Disclosure of Invention

The invention aims to: in order to solve the technical problems in reality, the salt hydrolysis method is provided for preparing crude heparin sodium by utilizing sow intestines.

The technical scheme adopted by the invention is as follows:

(1) intestinal mucosa extraction

Rolling healthy fresh sow intestines with deionized water by using a rolling machine for multiple times, then feeding the rolled intestines into an intestine scraping machine for scraping, and collecting intestinal mucosa mixed liquor prepared in the rolling and scraping processes;

(2) salt decomposition

Pumping the intestinal mucosa mixed solution into a salt decomposition tank by using a lift pump, stirring at normal temperature, adjusting the salinity of the mixed solution to 4.2 +/-0.3 ℃ by using saturated concentrated brine, adjusting the pH to 9.0 by using a sodium hydroxide solution, continuously heating to 58 +/-1 ℃, then adding trypsin, and carrying out heat preservation and deproteinization to ensure that the protein in the salt decomposition solution floats on the liquid surface after being solidified;

(3) separation of

Respectively separating oil and protein from the salinized solution through an intestinal residue filter barrel and a high-speed liquid-solid three-phase separator to obtain the salinized solution after protein and oil are separated, and pouring the salinized solution into a buffer tank;

(4) temperature reduction

Cooling the salt solution by using a plate heat exchanger to obtain an adsorption solution;

(5) adsorption

The adsorption solution is pumped into an adsorption tank, the pH value is adjusted to be more than 8.0, the salinity is more than 3.0, the treated resin is added for ion adsorption, and the adsorption is accelerated by stirring;

(6) resin collection

Collecting the adsorbed resin by a resin collecting tank, and pumping the resin into a rinsing tank for thorough cleaning to remove impurities such as protein, grease and the like;

(7) curing

And eluting, precipitating and drying the collected resin by using a traditional process to obtain a crude product of heparin sodium.

Wherein the water amount of the deionized water used in the step (1) is controlled to be 5 +/-1 Kg per sow intestine.

Wherein the stirring speed used in the step (2) is a speed of 60 +/-5 rpm.

Wherein the concentration of the sodium hydroxide solution used in the steps (2) and (5) is 0.3 +/-0.2 mol/L.

Wherein the method comprises the steps of:

preparing an enzyme inactivation solution;

(2) one-time heat preservation

Keeping the temperature of the mixture at 58 + -1 deg.C, pH 8.5, salinity > 3.1, and maintaining for 120 min;

(2) high temperature inactivation

Rapidly heating to 96 + -1 deg.C, adjusting pH to 8.3 + -1, and maintaining for 10min to make protein in the salt solution float on the liquid surface after fixing.

Wherein the method comprises the steps of:

preparing a salinization solution;

(1) rough fraction

Performing solid-liquid separation on the salinization liquid by using an intestinal residue filtering roller, and enabling the separated salinization liquid to enter a buffer tank;

(2) finely dividing

And (3) performing oil and protein fine separation on the coarsely separated salinized solution again by using a high-speed liquid-solid three-phase separator with the separation factor of 12200 +/-200 g and the rpm of 6400 +/-300 to obtain a second salinized solution.

Wherein the temperature of the step (4) is controlled to be 60 +/-2 ℃.

Wherein the resin in the step (5) is Bayer resin, and the dosage of the resin is 1Kg of Bayer resin added per 60Kg of small intestine of a pig.

Wherein the stirring speed in the step (5) is 55 +/-5 rpm, and the stirring time is 420 +/-60 min.

In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:

1. in the invention, because deionized water is adopted in the step (1) to extract and mix the intestinal mucosa of the sow, the reaction between ions in water and the effective component heparin sodium is reduced, and the concentration of the heparin sodium in the extracted intestinal mucosa mixed solution is improved.

2. In the invention, because the temperature of the mixed liquor is continuously raised to 58 +/-1 ℃ in the step (2), the mixed liquor is adjusted to the salinity of 4.5 ℃ and the pH is adjusted to 9.0, and the mixed liquor is stirred at the speed of 60 +/-5 rpm, so that the salting-out reaction effect of the trypsin is improved.

3. In the invention, because the heat preservation and high-temperature inactivation steps are adopted in the step (2), different enzymes can be subjected to salt decomposition and then inactivation treatment on different proteins, so that the decomposition effect on impurities such as other proteins is improved

4. In the invention, as the intestinal residue filtering drum and the high-speed liquid-solid three-phase separator are respectively adopted in the step (3) to carry out coarse separation and fine separation on the enzyme inactivation liquid, the protein and the grease can be separated, so as to improve the purification effect of the heparin sodium.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

In the description of the present invention, it should be noted that the terms "first", "second", and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance; furthermore, unless expressly stated or limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, as they may be fixedly connected, detachably connected, or integrally connected, for example; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.

Example (b):

a. rolling healthy fresh sow intestines with deionized water by using a rolling machine for multiple times, then feeding the rolled intestines into a intestine scraping machine for scraping, and collecting intestinal mucosa mixed liquor prepared in the rolling and scraping processes, wherein the water amount is controlled to be 5 +/-1 Kg per sow intestine;

b. pumping the intestinal mucosa mixed solution into a salt decomposition tank by using a lifting pump, stirring at the speed of 60 +/-5 rpm at normal temperature, adjusting the salinity of the mixed solution to 4.2 +/-0.3 ℃ by using saturated concentrated saline, adjusting the pH to 9.0 ℃ by using 0.3 +/-0.2 mol/L sodium hydroxide solution, and continuously heating to 58 +/-1 ℃;

c. keeping the temperature of the mixed solution at 58 +/-1 ℃, the pH value of 8.5 and the salinity of more than 3.1, preserving heat for 120min, quickly heating the mixed solution to 96 +/-1 ℃, adjusting the pH value to 8.3 +/-1, and preserving heat for 10min to ensure that the protein in the salinized solution floats on the liquid surface after being solidified;

d. performing solid-liquid separation on the salinized liquid by using an intestinal residue filtering roller, enabling the separated salinized liquid to enter a buffer tank to obtain a first salinized liquid, and performing oil and protein fine separation on the coarsely separated first salinized liquid again by using a high-speed liquid-solid three-phase separator with the separation factors of 12200 +/-200 g and the rpm of 6400 +/-300 to obtain a second salinized liquid;

e. cooling the second salinization solution by using a plate heat exchanger, and controlling the temperature to be 60 +/-2 ℃ to obtain an adsorption solution;

f. pumping the adsorption solution into an adsorption tank, adjusting the pH to be more than 8.0 by using 0.3 +/-0.2 mol/L sodium hydroxide solution, adjusting the salinity of the mixed solution to be more than 3.0 ℃ by using saturated concentrated brine, adding 1Kg of Bayer resin into each 60Kg of sow intestines for ion adsorption, and stirring and adsorbing at the speed of 55 +/-5 rpm for 420 +/-60 min;

g. collecting the adsorbed resin by a resin collecting tank, pumping the resin into a rinsing tank for thorough cleaning, removing impurities such as protein, grease and the like, and discharging the adsorption waste liquid into sewage treatment;

h. and eluting, precipitating and drying the collected resin by using a traditional process to obtain a crude product of heparin sodium.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.