KR20170029328A

KR20170029328A - Extraction method of collagens from plant

Info

Publication number: KR20170029328A
Application number: KR1020150126529A
Authority: KR
Inventors: 조환기
Original assignee: 청양포포나무농장영농조합법인
Priority date: 2015-09-07
Filing date: 2015-09-07
Publication date: 2017-03-15
Also published as: KR101783469B1

Abstract

The present invention relates to a method for extracting collagen from a gold coin, comprising: (a) dispersing in a alkaline aqueous solution having a pH of 9 to 13 any one selected from the group consisting of roots, leaves, stems, flowers, ; And (b) adding an acid after a predetermined time elapses after the step (a) to neutralize the solution to a pH of 6 to 8 to elute the soluble vegetable collagen. According to the present invention as described above, it is possible to extract vegetable collagen easily from a gold coin, thereby solving the weak points of animal collagen and contributing to the localization of vegetable collagen which depends on import alone.

Description

Extraction method of collagen from plant]

The present invention relates to a method for extracting vegetable collagen, and more particularly, to a method for extracting collagen from a gold coin.

Collagen is the most abundant fibrous protein in the animal's body. It is the main component of the skin's dermis and connective tissue, and 90% of the proteins that make up the bone are collagen. Collagen, which is generally used for medical and cosmetic purposes, And a polymer protein consisting of three spiral chains of polypeptide chains of about 100,000 molecular weight extracted from the skin.

The molecular weight of collagen is approximately 130,000 and is a Polypeptide that constitutes one third of the mammalian tissue (skin, connective tissue, and the major component of organic matter in bone and teeth). It is produced by dissolving mineral parts of bone with phosphoric acid.

Isolation from tissues is obtained by organic solvent extraction, acid alkali treatment, and trypsin hyaluronidase treatment to give insoluble collagen. The constituent amino acid is proline oxyproline (hydroxyproline) glycine glutamic acid and the like. Among them, the content of oxyproline which is not present in other proteins is high.

These nutritional values of collagen are as follows: 1) Reinforcement of proteins such as health supplements, functional beverages, sports nutrition, and weight control foods 2) Effect and value in dietary fat, sugar, and salt restriction 3) It is known that it acts as an effective carrier of active ingredients such as aroma 4) It acts as a joint protecting agent by an ideal nutritional supply for degenerative joint disease 5) It promotes growth by nutrition of hair and nails 6) It strengthens and protects connective tissues.

Recently, the aging delay has been actively studied due to the rapid increase of the elderly population, and the craving for youth is hardly possible for all ages. In this age of boom, the efficacy and effectiveness of collagen has increased, and the number of people looking for collagen is increasing.

However, most of the collagen currently in circulation is an animal collagen produced by selecting raw materials from cattle, and it is anxious due to foot-and-mouth disease and mad cow disease. Accordingly, the development of vegetable protein (hereinafter referred to as "vegetable collagen") containing the main components of collagen, such as Hydroxy lysine and Hydroxy lysine (hereinafter referred to as "vegetable collagen"), There is a need to supplement the above.

Currently, vegetable collagen is not produced in Korea at present, but it is produced by some foreign companies. However, since the products extracted from foreign countries are extracted by using a chemical called glycol as a solvent, they are used only in cosmetics. However, So that the use thereof is avoided.

1. Korean Patent Publication No. 10-2012-0134935 2. Korean Patent Publication 10-2011-0028889

Accordingly, an object of the present invention is to establish a water-soluble vegetable collagen production method using gold leaf which is one of the plants in a hibiscus plant material, thereby solving the weak points of animal collagen and localizing the collagen of vegetable.

In order to achieve the above object,

(a) dispersing any one selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof in an alkaline aqueous solution having a pH of 9 to 13; And

(b) adding an acid after a predetermined time elapses after the step (a) to neutralize the solution to a pH of 6 to 8 to elute the water soluble vegetable collagen; and do.

According to the present invention, there is provided a method for extracting vegetable collagen from a gold coin according to the present invention, which comprises the steps of: (a) extracting a vegetable collagen from a gold coin, which is selected from the group consisting of roots, leaves, May be a gold-plated granule made by mixing powder selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof and a binder.

In the method of extracting vegetable collagen from gold coin according to the present invention as described above, the predetermined time of step (b) may be 1 to 6 hours.

According to the present invention, there is further provided a method of extracting vegetable collagen from a gold coin according to the present invention, the method further comprising a centrifugal separation step of centrifuging the product of step (b) to isolate a layer containing vegetable collagen .

In the method of extracting vegetable collagen from gold coin according to the present invention as described above, it is preferable that any one selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof is a gold coin Do.

According to the present invention as described above, it is possible to extract vegetable collagen easily from a gold coin, thereby solving the weak points of animal collagen and contributing to the localization of vegetable collagen which depends on import alone.

FIG. 1 is a graph showing the cell proliferation ability of the extract prepared according to the present invention.

Hereinafter, the present invention will be described in detail.

The present invention relates to a method for extracting vegetable collagen from a gold coin, wherein the gold coin is an annual herbaceous plant belonging to the genus Hibiscus, which is also called a wild bouillon, . Recently, seeds have been introduced from China into Korea and cultivation has been carried out in Korea.

In July to August, the gold coins grow in excess of the height of a man. The diameter of the blossoms is usually 16 to 18 cm. The petals are yellow, thin and scarlet, and the leaves are very beautiful, with dark blue leaves. It is said that it is very important to harvest on time because it is only about six hours to bloom.

The flower of the gold coin is cut from the end of July and is torn and dried. Cosmetics and tea are used as raw materials, and gold leaf is also used for various purposes because it contains various physiologically active substances.

Hereinafter, a process for extracting vegetable collagen from a gold coin will be described in detail.

The method according to the present invention for extracting vegetable collagen from a gold leaf is carried out by dispersing any one selected from the group consisting of roots, leaves, stems, flowers and mixtures thereof in an alkaline aqueous solution having a pH of 9 to 13 Adding the acid to the mixture to neutralize the mixture to a pH of 6 to 8, and eluting the soluble vegetable collagen.

In this case, inorganic alkali such as sodium hydroxide solution or potassium hydroxide aqueous solution having a concentration of 0.1 to 10% is used as alkaline aqueous solution, and ammonia and amine are not used because chemical properties (skin and food use can not be used) are changed.

In addition, it can be boiled or heat treated before neutralization, and the temperature of the solution should be 60 ° C to 90 ° C during warming. It should be cooled in a short time after completion of the warming operation within 1 hour. In case of neutralization, various inorganic acids (eg hydrochloric acid, Organic acids (eg, citric acid, malic acid, etc.) can be used.

The process of decomposing plant tissues using alkaline is a process that destroys plant tissues and prevents water-soluble components from remaining in plant tissues.

In the present invention, it is possible to use all the outposts of gold coins, or to extract vegetable collagen by using any one of them or by using a mixture of various combinations. Preferably, flowers of gold coins are used.

In addition, the extraction time is not particularly limited, but it has been determined that the extraction time is sufficient for about 6 hours by one of the various tests of the present inventors. If the extraction time is less than 1 hour, sufficient time for extraction can not be secured. There is a problem in that the extraction time alone becomes unnecessarily long without increasing the efficiency.

In order to reduce the extraction time and increase the extraction efficiency, the present invention can be used as a granule of gold coins, which will be described in detail.

The powder of any one selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof is prepared as a powder, and the powder is mixed with a binder to produce gold and silver granules.

Such gold coins may be dried and used as powders. In the present invention, instead of extracting directly with powder, gold coins are prepared by mixing with a binder and then extracted with collagen.

As described above, the reason for manufacturing the gold granules is that when the powder is directly introduced into the extraction solvent, the disintegration of the gold powder is lowered and the specific gravity of the powder is smaller than that of the extraction solvent, This causes problems that can not be extracted very well apart.

Especially, in the case of gold coin, most of the viscous collagen is a water-soluble substance to be extracted, so that it is not dispersed even more.

In order to solve this problem, in order to solve this problem, a granule which is widely used in a pharmaceutical composition and a powder of gold powder is mixed to make granules, and the specific gravity of the granules is made higher than that of the extraction solvent, And the plant collagen extraction is facilitated by contacting the gold powder with water while gradually dissolving the binder in the extraction solvent and making contact with the extraction solvent.

The binder can be used without any particular limitation as long as it is widely known to make granules. Examples of the binder include lactose, anhydrous lactose, spray dried lactose, sugar, povidone, methylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose Cellulose, hydroxyethylcellulose, eudragit, gelatin, luocom, xanthan com, rebaudioside A, xylitol, maltose, maltitol, mannitol, palatinose, erythritol, trehalose, glucose, sorbitol, maltodextrin, dextrose , Oligosaccharide hydroxypropylmethylcellulose, carboxymethylcellulose sodium, carboxymethylcellulose calcium, microcrystalline cellulose, low substituted hydroxypropylcellulose, corn starch and the like, camellose calcium, pregelatinized starch, crospovidone, low substitution Hydroxypropylcellulose, glycolic acid Sodium, co-povidone, carbomer, carboxymethylcellulose, croscarmellose sodium and the like, and the content thereof is such that when it is enough that the extracting solvent can sink to a certain degree when the gold granule is made, , It takes too much time to release the bond in the extraction solvent and the extraction time becomes long

The method of producing the granules may be any of the direct method, dry granulation and wet granulation. However, it is easy to manufacture the granules by using the wet granulation method in view of the characteristics of the gold granules and when the wet granulation method is used, It is easy to control.

A simple description of wet granule manufacturing method is as follows. After mixing the gold powder and the binder, the mixture is homogenized in a certain container, and the homogenized mixture is sprayed with water free of charge. The free-sprayed water gives the binder a viscosity, and the water-soluble substance, which is absorbed by the surface of the gold powder, is eluted and added with the action of the binder, so that the binding force can be strengthened. The granules thus produced are dried and used.

In this case, the specific gravity is not so large that the dispersion exhibits the same physical behavior as in the case of not being made into a granule in the subsequent extraction solvent, thereby improving the dispersibility. Do not.

The weight of the binder is preferably 50 parts by weight to 150 parts by weight with respect to 100 parts by weight of the powdery copper powder when preparing the granules as described above, and when the amount is 50 parts by weight or less, the specific gravity is small, In the case of 150 parts by weight, the binding force is too strong and the extraction time is long.

In order to solve the disadvantage that the extraction time is prolonged in the present invention, granules can be prepared by further including a disintegrant used for preparing granules or tablets. The disintegration is intended to allow the granules to quickly dissolve in the extraction solvent to improve contact with the extraction solvent.

As long as it is widely known as disintegration in the technical field to which the present invention belongs, it can be used without any particular limitation, and it is suitable to use 5 to 20 parts by weight with respect to 100 parts by weight of gold powder.

Although extraction of vegetable collagen is completed as described above, only a layer containing vegetable collagen is separated from the extraction stock solution containing gold powder and granules thereof using a centrifugal separator to obtain only the vegetable collagen layer When the step of concentrating vegetable collagen is carried out, the extraction method according to the present invention is completed.

The conditions and methods of centrifugation are well known in the art to which the present invention belongs, and can be carried out without particular limitations as long as those skilled in the art are familiar with the present invention.

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

< Example 1>

100 g of the outpowder including gold leaf roots was placed in an aqueous solution of sodium hydroxide of pH 10, and the mixture was slowly stirred. After 5 hours, the citric acid aqueous solution was added to neutralize the mixture to pH 7, followed by centrifugation at 1000 rpm for 30 minutes Lt; / RTI > to separate the layer containing vegetable collagen.

< Test Example 1: Analysis of amino acids in extracts>

Total amino acid composition and OH-Proline analysis were applied to total amino acid analysis. For extraction and pretreatment, 100 μL of the sample was subjected to hydrolysis and PITC labeling using the PICO-tag method, and the PITC labeled samples 20 μL was taken out from 400 μL and loaded on HPLC to obtain chromatogram. The assay conditions were a reaction amount of 100 μL or mg, an injection volume of 50 μL, a PITC-labeling volume of 400 μL, and an injection amount of 12.5 μL.

Hydroxyproline (Hydroxyproline) and Hydroxylysine (Hydroxy lysine) were abundantly contained as 11.81 / 100g and confirmed as vegetable collagen. In addition, glutamic acid (glutamic acid Glutamic acid, aspartic acid, and arginine.

< Test Example 2: Evaluation of wrinkle-improving effect>

The cell efficacy of the wrinkle-improving effect on the collagen prepared in Example 1 was examined. The extract of Example PE (Patent Extract) was used as a reference material and the comparative raw materials were CP (commercial product) and OO Biotech Co., CE (Carrot Extract) and FE (Flammulina Velutipes Extract), which were developed by the Ministry of Health, Labor and Welfare.

The MTT assay method was applied to the test method, and the results are shown in Fig. As shown in FIG. 1, the cell proliferation ability was about 0.001% to 20.5%, CP 14.2%, CE 17.6%, FE 15.2%, and the extract according to the present invention had the highest value among the four raw materials Could know.

< Example 2>

100 g of the powder of the outpouring powder including the roots of the gold coins were placed in a wet granulator (FY-KS-2) homogenously mixed with 100 g of lactose to prepare gold coin granules. The granules were placed in an aqueous solution of sodium hydroxide at pH 10, and the granules were waited until the granules were sufficiently submerged. The granules were completely loosened and waited until the extraction was sufficiently completed. After 2 hours, an aqueous solution of citric acid was added to neutralize the mixture to pH 7 The mixture was placed in a centrifuge and stirred at 1000 rpm for 30 minutes to separate the layer containing vegetable collagen.

As a result of measuring the weight after separating the layer, the weight was found to be about 20% higher than that in Example 1, and it was confirmed that the granules were well extracted.

< Example 3>

(FY-KS-2) prepared by homogenously mixing 90 g of lactose and 10 g of corn starch into 100 g of the powder of the outpouring powder containing the gold leaf, The granules were placed in an aqueous solution of sodium hydroxide at pH 10, and the granules were slowly stirred until the granules were sufficiently submerged. While the granules were completely loosened, the time required until the extraction was sufficiently completed was 15 minutes And it was confirmed that the total extraction time was 1 hour and 15 minutes, and the extraction time could be shortened more than Example 2.

< Example 4>

100 g of the powder of the kinkakujisu was added to a wet granulation machine (FY-KS-2) homogeneously mixed with 100 g of lactose, to prepare a gold coin granule. The granules were placed in an aqueous solution of sodium hydroxide of pH 10, and the granules were waited until the granules were sufficiently submerged. The granules were completely loosened and the granules were completely loosened until the extraction was sufficiently completed. After 5 hours, an aqueous citric acid solution was added to neutralize the mixture to pH 7 The mixture was placed in a centrifuge and stirred at 1000 rpm for 30 minutes to separate the layer containing vegetable collagen.

As a result of weighing after separating the layers, the same weights as in Example 2 were used, but the weights thereof were measured to be about 15% higher, and it was confirmed that the extract was further extracted from the flower powder.

Claims

(a) dispersing any one selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof in an alkaline aqueous solution having a pH of 9 to 13; And
(b) adding an acid after a predetermined time elapses after the step (a) to neutralize the solution to a pH of 6 to 8 to elute the soluble vegetable collagen.

The method according to claim 1, wherein any one selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof is dispersed in step (a)
A method for extracting vegetable collagen from gold coins obtained by pulverizing any one selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof, .

The method of extracting vegetable collagen from a gold coin according to claim 2, wherein the gold coarse granule further comprises a disintegrant.

The method of any one of claims 1, 2, and 3, wherein the predetermined time of step (b) is 1 to 6 hours.

The method according to any one of claims 1, 2, and 3, further comprising a centrifugal separation step of separating the layer containing plant collagen by centrifuging the result of step (b) A method of extracting vegetable collagen from a gold coin.

The method according to any one of claims 1, 2, and 3, wherein the gold leaf is selected from the group consisting of roots, leaves, stems, flowers, and mixtures thereof, A method for extracting vegetable collagen from gypsum.