KR20010070901A - manufacturing method for chondroitin sulfate and its content - Google Patents

Abstract

PURPOSE: A chondroitin sulfate compound and a method for producing material containing the compound are provided to obtain the compound and composition thereof useable as raw materials of medicine, cosmetic, commercial food, health food and health aid food by extracting the compound from cartilage of cattle and purifying it into the resultant compound. CONSTITUTION: The chondroitin sulfate compound is prepared by a first process of removing impurities from cartilage of cattle by immersing the cartilage into a diluted hydrochloric acid solution; a second process of heating the obtained material in a pressure container under pressure; a third process of extracting a portion containing chondroitin sulfate component by adding any protease; a fourth process of removing proteins excluding chondroitin sulfate by regulating pH value of the extract; a fifth process of purifying the obtained material by removing ionic salts and amines through ion-exchange resin; and a sixth process of separating chondroitin sulfate and its salt with alcohol.

Description

Manufacturing method for chondroitin sulfate and its content

The present invention relates to a chondroitin sulfate and a method for producing the same, and more particularly, chondroitin sulfate to extract the chondroitin sulfuric acid, the main component of the animal connective tissue from the small cartilage to purify the high purity and to produce the contents at the same time and It relates to a method for producing the contents.

The current life expectancy of humans has been extended, but in addition, patients suffering from degenerative diseases such as osteoporosis, rheumatism, arthritis and deformable arthritis are also increasing.

Chondroitin sulfate is known to be effective in preventing, relieving and treating degenerative diseases caused by aging, brain cell necrosis and dry eye.

Conventionally, the chondroitin sulfate is extracted from 0.15M potassium chloride or 4.0M guanidinium chloride solution in 50mM sodium acetate buffer, using shark cartilage or snail as a raw material, and then centrifuged to remove salt. After dialysis, it was prepared by the method of freeze-drying, but the extraction method is complicated, and a facility such as a dialysis apparatus for removing salts is required, and there is a problem that it is difficult to purify the high purity and high purity.

In addition, chondroitin sulfate prepared as described above also has a problem in that almost all depend on imports.

Accordingly, the present invention has been made to solve the above problems, extract chondroitin sulfate from bovine cartilage and purify with high purity chondroitin sulfate, and at the same time to produce the contents of pharmaceuticals, cosmetic raw materials, general food raw materials, An object of the present invention is to provide a method for producing chondroitin sulfate and its contents which can be used as raw materials for health foods and health supplements.

1 is a process diagram showing a process for producing chondroitin sulfate and its contents according to the present invention.

The present invention for achieving the above object, the impurity removal process to remove impurities by dipping small cartilage in dilute hydrochloric acid solution; A pressurized heating process of putting the small cartilage that has undergone the impurity removal process into a pressurized container and heating it under a pressurized state; An extraction process of adding a protease to extract an extract containing chondroitin sulfate component from cartilage; Adjusting the pH of the extract to remove proteins other than chondroitin sulfate; Purifying the extract using ion exchange resin to remove ionic salts and amines; In addition, a separation process for separating chondroitin sulfuric acid and its salts using an alcohol; and a method for producing chondroitin sulfate and its contents comprising a technical gist.

Here, the small cartilage that has undergone the impurities removal process is preferably washed with ethanol.

Then, the pH of the extract is neutralized with caustic soda solution after the purification process.

In addition, the ion exchange resin used in the purification process is a cation exchange resin and an anion exchange resin is mixed, the alcohol used in the separation process includes at least one or more of the group consisting of ethanol, methanol, propyl alcohol, butyl alcohol. Is preferably configured.

Therefore, chondroitin sulfate can be extracted from bovine cartilage, refined into high purity chondroitin sulfate, and the contents thereof can be prepared to be used as raw materials for medicines, cosmetic raw materials, general food raw materials, health foods and health supplements. There is an advantage in that sulfuric acid and its contents are prepared.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

1 is a process chart showing a process for producing chondroitin sulfate and its contents according to the present invention.

As shown, chondroitin sulfate and a method for producing the same according to the present invention is largely composed of an impurity removal process, pressurized heating process, extraction process, protein removal process, purification process, pH neutralization process and separation process do.

First, the impurity removal process will be described.

The impurity removal process is a process of removing blood and other impurities contained in the small cartilage, and the small cartilage is immersed in dilute hydrochloric acid solution having a pH of about 1.0 to 6.0 to remove the blood and ash contained in the small cartilage. At this time, the immersion time is suitable for 3 to 30 hours, the shorter the better for the productivity improvement, but if the time is too short, the removal rate of the ash is reduced.

Table 1 shows the relationship between the dipping time of dilute hydrochloric acid solution and ash content.

As shown in Table 1, as the immersion time increases, the content of residual ash decreases. In view of the above point, the most suitable time is preferably about 18 hours.

The small cartilage, which has undergone the impurity removal process by dipping in dilute hydrochloric acid solution, is washed with 95% alcohol to remove other impurities.

Bovine cartilage washed with alcohol is used as a raw material for the next step.

The alcoholic cartilage bovine cartilage is a process for extracting chondroitin sulfate which is strongly bound to collagen from the barium cartilage.

In the pressure heating process, the cartilage is put into a pressure cooker, and then heated to 100 ° C to 130 ° C to extract the extract.

Pressurization time is made in about 1 hour-about 6 hours.

Table 2 shows the pressurized heating extraction time and the yield of chondroitin sulfate.

As shown in Table 2, the yield increased with increasing pressurization time. However, considering the production cost and productivity, it is preferable to heat the pressure for about 3 hours.

Next, an extraction process for re-isolating chondroitin sulfate, which binds to a protein such as collagen, is performed using the extract extracted in the pressurization heating process.

In the extraction process, chondroitin sulfate is separated by hydrolysis by adding 0.1% to 0.5% of a protease including alcarase, neutrase, pepsin, papain, etc. to the extract extracted in the pressurized heating process. Serve

Table 3 shows the hydrolysis time and the yield of chondroitin sulfate.

As shown in Table 3, it can be seen that as the hydrolysis time increases, the yield of chondroitin sulfate also increases. In case of extracting for more than 12 hours, the yield increased to a certain degree, but the decomposition resulted in denaturation of the extract, which lost its value as a final product.

Therefore, in consideration of productivity, it was found that it is most suitable to hydrolyze about 4 hours when the increase in yield is the maximum.

Residues such as collagen and undecomposed protein are still present in the extract that has undergone the extraction process. In order to remove residual impurities, a protein removal process proceeds to the next process.

The protein removal process is carried out by adding a sodium hydroxide solution to the extract extracted in the extraction process, by adjusting the pH to 7.5 ~ 9.0 to precipitate and remove impurities by filtration.

The extraction process of the protein removal process is a purification process, the purification process is a process of removing the salts and amines contained in the extract by using an ion exchange resin.

That is, in the extract, many salts and protein decomposition products exist in the form of amines in addition to chondroitin sulfate, so that cationic salts and amines are adsorbed using a cation exchange resin to purify chondroitin sulfate of high purity, and anion is used by using an anion exchange resin. The salts are adsorbed and removed.

Ion-exchange resins used in the purification process used commercially available resins of porous polymer materials composed of copolymers of styrene and divinylbenzene. In the case of cation exchange resin, the activation of the ion exchange resin was passed through hydrochloric acid solution and washed with purified water, and the anion exchange resin was passed through sodium hydroxide solution and washed with purified water.

The activated ion exchange resin was used by mixing a cation exchange resin and an anion exchange resin.

Table 4 is a table showing the volume ratio of the cation exchange resin and anion exchange resin and the yield of chondroitin sulfate.

As shown in Table 4, the yield of chondroitin sulfate was the best when the volume ratio of cation exchange resin: anion exchange resin was 9: 1.

After the above purification process, high purity chondroitin sulfate is present in the extract, and a pH neutralization process is performed to increase solubility in water.

The pH neutralization process is carried out by neutralizing the extract with a caustic soda solution, and after the above process, the solubility of the extract is increased to increase the solubility of water in pharmaceuticals, cosmetic raw materials, general food raw materials, health food and health It is in a state suitable for use as a raw material for supplements.

Next, a separation process for separating chondroitin sulfate is performed.

In the separation process, the neutralized extract is precipitated with chondroitin sulfate using ethanol, the precipitated chondroitin sulfate is recovered, washed with ethanol and dried to obtain chondroitin sulfate.

Although ethanol is used in the separation process, one or a mixture of two or more of methanol, propyl alcohol, butyl alcohol, and isomers thereof may be used.

The following describes a specific example of extracting high purity chondroitin sulfate.

<Example 1>

500 g of bovine cartilage, which was subjected to impurity treatment using dilute hydrochloric acid solution, was added to 5000 ml of purified water, and then extracted by pressurized heating at 121 ° C. for 3 hours. Hydrolysis to a degree. It was adjusted to pH 8.5, and after about 1 hour, it was confirmed that a precipitate was formed and filtered using diatomaceous earth. The filtrate was mixed with a cation exchange resin and an anion exchange resin at 9: 1 and eluted at a flow rate of 50 ml per minute in a packed resin column to neutralize the eluate with caustic soda.

The precipitated chondroitin sulfate was recovered using ethanol, washed with ethanol, and dried to extract about 13 g of white chondroitin sulfate.

<Example 2>

Except for using methanol instead of ethanol through the same process as in Example 1 was extracted white chondroitin sulfate of about 13g.

<Example 3>

Except for using propyl alcohol instead of ethanol through the same process as in Example 1 was extracted white chondroitin sulfate of about 13g.

<Example 4>

Except that butyl alcohol was used instead of ethanol, 13 g of white chondroitin sulfate was extracted through the same process as in Example 1.

<Example 5>

The proteolytic enzyme was extracted with about 13.5 g of white chondroitin sulfate through the same process as in Example 1 except that nutrase was used instead of alkalase.

<Example 6>

Except for using methanol instead of ethanol through the same process as in Example 5 was extracted white chondroitin sulfate of about 13.5g.

<Example 7>

Except that propyl alcohol was used instead of ethanol, white chondroitin sulfate of about 13.5 g was extracted through the same process as in Example 5.

<Example 8>

Except that butyl alcohol was used instead of ethanol, white chondroitin sulfate of about 13.5 g was extracted through the same process as in Example 5.

<Example 9>

500 g of bovine cartilage, which was subjected to impurity treatment using dilute hydrochloric acid solution, was added to 5000 ml of purified water, and extracted by pressurized heating at 121 ° C. for 3 hours. Then, the extract was adjusted to pH 3.0 and temperature of 50 ° C., and 0.3% of pepsin was added. Disassembled. It was adjusted to pH 8.5, and after about 1 hour, it was confirmed that a precipitate was formed and filtered using diatomaceous earth. The filtrate was mixed with a cation exchange resin and an anion exchange resin at 9: 1 and eluted at a flow rate of 50 ml per minute in a packed resin column to neutralize the eluate with caustic soda.

The precipitated chondroitin sulfate was recovered using ethanol, washed with ethanol, and dried to extract about 13.2 g of white chondroitin sulfate.

<Example 10>

13.2 g of white chondroitin sulfate was extracted by the same process as Example 9 except that methanol was used instead of ethanol.

<Example 11>

Except for using propyl alcohol instead of ethanol through the same procedure as in Example 9 was extracted white chondroitin sulfate of about 13.2g.

<Example 12>

Except for using butyl alcohol instead of ethanol through the same process as in Example 9 was extracted about 13.2g of white chondroitin sulfate.

<Example 13>

500 g of bovine cartilage, which was subjected to impurity treatment using dilute hydrochloric acid solution, was added to 5000 ml of purified water, and extracted by pressurized heating at 121 ° C. for 3 hours. Then, the extract was adjusted to pH 7.0 and temperature of 50 ° C. and 0.3% of trypsin was added. Disassembled. It was adjusted to pH 8.5, and after about 1 hour, it was confirmed that a precipitate was formed and filtered using diatomaceous earth. The filtrate was mixed with a cation exchange resin and an anion exchange resin at 9: 1 and eluted at a flow rate of 50 ml per minute in a packed resin column to neutralize the eluate with caustic soda.

The precipitated chondroitin sulfate was recovered using ethanol, washed with ethanol, and dried to extract about 14 g of white chondroitin sulfate.

<Example 14>

About 14 g of white chondroitin sulfate was extracted through the same process as in Example 13 except that methanol was used instead of ethanol.

<Example 15>

Except for using propyl alcohol instead of ethanol through the same process as in Example 13 was extracted about 14g of white chondroitin sulfate.

<Example 16>

Except for using butyl alcohol instead of ethanol through the same process as in Example 13 was extracted about 14g of white chondroitin sulfate.

<Example 17>

500 g of bovine cartilage, which was subjected to impurity treatment using dilute hydrochloric acid solution, was added to 5000 ml of purified water, and then extracted by pressurized heating at 121 ° C. for 3 hours. Disassembled. It was adjusted to pH 8.5, and after about 1 hour, it was confirmed that a precipitate was formed and filtered using diatomaceous earth. The filtrate was mixed with a cation exchange resin and an anion exchange resin at 9: 1 and eluted in a packed resin column at a flow rate of 50 ml per minute to neutralize the eluate with caustic soda.

The precipitated chondroitin sulfate was recovered using ethanol, washed with ethanol, and dried to extract about 13.5 g of white chondroitin sulfate.

<Example 18>

Except for using methanol instead of ethanol through the same process as in Example 17 was extracted white chondroitin sulfate of about 13.5g.

<Example 19>

Except for using propyl alcohol instead of ethanol through the same procedure as in Example 17 was extracted white chondroitin sulfate of about 13.5g.

<Example 20>

Except that butyl alcohol was used instead of ethanol, white chondroitin sulfate of about 13.5 g was extracted through the same process as in Example 17.

The present invention by the above configuration has the effect of supplying raw materials and mass production is possible by extracting chondroitin sulfate from bovine cartilage and purifying with high purity chondroitin sulfate.

In addition, since high-purity chondroitin sulfate is manufactured, it can also be used as a raw material for medicines, cosmetic raw materials, general food raw materials, health foods and health supplements.

Claims (5)

An impurity removal process for removing impurities by dipping small cartilage in dilute hydrochloric acid solution; A pressurized heating process of putting the small cartilage that has undergone the impurity removal process into a pressurized container and heating it under a pressurized state; An extraction process of adding a protease to extract an extract containing chondroitin sulfate component from cartilage; Adjusting the pH of the extract to remove proteins other than chondroitin sulfate; Purifying the extract using ion exchange resin to remove ionic salts and amines; And, Separation process for separating chondroitin sulfuric acid and its salts using alcohol; Chondroitin sulfuric acid and its production method comprising a. According to claim 1, Chondroitin sulfate and a method for producing the same, characterized in that the cartilage after the impurity removal process is washed with ethanol. 2. The method of claim 1, wherein the pH of the extract is neutralized with caustic soda solution after the purification process. The method of claim 1, wherein the ion exchange resin used in the purification process is mixed with a cation exchange resin and an anion exchange resin. The method of claim 1, wherein the alcohol used in the separation process comprises at least one or more of the group consisting of ethanol, methanol, propyl alcohol, and butyl alcohol.