KR101362800B1 - Prolamine-positive ion complex contaning phytosterol and method thereof - Google Patents

Abstract

The present invention relates to a prolamine-positive ion polymer complex containing phytosterol and a method thereof. According to an embodiment of a prolamine-positive ion polymer complex containing phytoster according to the present invention comprises phytosterol, prolamine, and a positive ion polymer. The phytosterol is stigmasterol, and the prolamine and the positive ion polymer are coating the phytosterol.

Description

PROLAMINE-POSITIVE ION COMPLEX CONTANING PHYTOSTEROL AND METHOD THEREOF}

The present invention relates to a prolamin-cation complex contained in phytosterol and a preparation method thereof.

For effective delivery, digestion, and absorption of water-soluble active ingredients in the body, the collected water-soluble active ingredients are preserved in the stomach with low pH and high acidic conditions, and the contents are stored in the intestine with a relatively long retention time. There is a need for a system that adjusts to release slowly. The system requires encapsulation, and this encapsulation technique is a technique for trapping a water-soluble active ingredient inside a substance or tissue so that a solid, liquid, or gaseous water-soluble active ingredient can be released at a controlled rate under specific conditions. Generally, synthetic or natural high molecular weight materials are used for the capsule.

On the other hand, chitosan is a natural polymer material obtained through deacetylation from chitin and has very low toxicity [J. Microencapsulation, 2000, 17, 625 ~ 638], it is easy to gel and form a film, and has a multivalent cationic property so that it can be easily combined with anionic materials [J. Chem. Technol. Biotechnol., 1990, 49, 395-404], and also has an excellent intestinal absorption-5-adhesion [J. Pharm. Sci., 2003, 92, 567 ~ 576], a lot of researches on the technology to effectively deliver drugs or functional substances through the encapsulation using chitosan. Capsule preparation method using chitosan include emulsion cross-linking [Int. J. Pharm., 1994, 11, 217-222], emulsion droplet coalescence [Pharm. Res., 1999, 16, 1830-1835], reverse micellar method [J. Controlled Release, 2001, 17, 317-323], spray-drying [Int.J. Pharm., 1994, 217-222), or an O / W / O multiple emulsion method (Korean Patent 10-2005-0014831) and the like are used. However, in the preparation of capsules using chitosan, if only chitosan is used, the preparation of capsules is not easy, and there are many ionic gelation methods for forming capsules in a short time through rapid crosslinking reaction between polyvalent cationic chitosan and crosslinking agents having counter ions. It is trying. In ionic gelation, an insoluble membrane is formed mainly by a crosslinking reaction on the polymer surface, and encapsulation is achieved through phase separation from the contents. The capsule thus prepared mainly consists of synthetic or natural polymers, and serves to control the release of the contents. At this time, the release control of the content is determined by the chemical structure, thickness and size of the capsule membrane and the concentration of the material constituting the capsule, the concentration of the content, the concentration of the crosslinking agent, and the pH [Int. J. Pharm., 2004, 274, 1-33, J. Controlled Release, 2004, 150, 5-28]. As the crosslinking agent added for the ionic gelation of chitosan, glutaraldehyde and sulfuric acid were mainly used [J. Microencapsulation, 1998, 15, 373-382; J. Microencapsulation, 2002, 19, 173-180; Korean Patent Nos. 10-1998-0056584, 10-2003-0085599]. However, these substances are toxic, making direct use in food difficult and limited to pharmaceuticals.

The present inventors have thus arrived at the present invention to prepare chitosan capsules which have high yield and high preservation in the stomach and are slowly released in the intestine.

An object of the present invention is to provide a composite comprising a phytosterol, a prolamine and a cationic polymer having a proper particle size, a stable zeta potential, and a constant elution.

In order to achieve the above object, in one embodiment of the present invention, a phytosterol, a prolamine, and a cationic polymer are included, and the prolamin and cationic polymers contain phytosterol, which is characterized by coating a phytosterol. Prolamin-cation polymer composite.

The "phytosterol" of the present invention includes, but is not limited to, sterol compounds derived from plants, including sterols and stanols in which sterols are hydrogenated. Representative sterols include β-sitosterol, stigmasterol, and campesterol, which are widely distributed in vegetable systems such as fruits, vegetables, nuts, grains, and legumes. Stanols include sitostanol and campestanol, which are found in small amounts in cereals and vegetable oils such as barley and corn. Phytosterols are known to inhibit cholesterol absorption, thereby reducing total cholesterol and LDL cholesterol levels in the blood. Plant sterols are sterols that are found in many plant foods. Cytosterols present in cottonseed and stigmasterols present in soybeans were known to exist only in the vegetable world and were named after them. On the other hand, cholesterol having 27 carbon atoms is called animal sterol, and carbon 28 ergosterol is called mycosterol (microbial sterol).

In the present invention, "prolamin" is selected from the group consisting of but not limited to zein, gliadin, hordein, secalin and kafirin It may be characterized in that any one or more, the prolamin may be characterized in that the zein (zein).

In the present invention, the "cationic polymer" may be characterized as chitosan, wherein the chitosan is poly [ß- (1-4) -2-amino-2-dioxy-D-glucopyranose], thiolated Chitosan, trimethylated chitosan, carboxymethyl chitosan and N- (2-hydroxyl propyl-3-trimethyl ammonium chitosan chloride may be any one or more selected from the group consisting of, the molecular weight of the cationic polymer is characterized in that 800-2000cp It may be, the concentration of the cationic polymer may be characterized in that 0.2 to 0.5%.

In one embodiment, in mixing phytosterol, prolamine and cationic polymer, the prolamin and cationic polymer coats phytosterol, preparing phytosterol-containing prolamin-cation polymer composite. It may provide a method.

In the above embodiment, the phytosterol is any one or more selected from the group consisting of β-sitosterol, stigmasterol, campesterol, sitostanol and campestanol. Provided is a prolamin-cationic polymer composite. Further, in the above embodiment, the prolamin is any one or more selected from the group consisting of zein, gliadin, hordein, secalin and kafirin. In particular, the prolamin may be zein, and the concentration of zein may be 15 mg / ml to 25 mg / ml. At this time, Jane is a protein extracted from corn is a kind of prolamin soluble in alcohol.

Also in this embodiment, the cationic polymer may be chitosan, wherein the chitosan is poly [ß- (1-4) -2-amino-2-dioxy-D-glucopyranose], thiolated chitosan, trimethylated Chitosan, carboxymethyl chitosan and N- (2-hydroxypropyl-3-trimethyl ammonium chitosan chloride may be any one or more selected from the group consisting of, the cationic polymer may be characterized in that the molecular weight of 800-2000cp, The concentration of the cationic polymer may be 0.3 to 0.7% The chitosan is a cationic polymer having an amino group which is a cation, and has a characteristic of strongly catching and releasing harmful substances with anions.

The present invention provides a complex for capturing phytosterol, it is possible to control the encapsulation efficiency, elution properties and the like by changing the molecular weight, concentration, etc. of chitosan included in the complex.

1 is a graph showing the particle size and zeta potential of the complex according to the concentration of zein when chitosan is not added.
Figure 2 is a graph showing the particle size and zeta potential of the complex according to the concentration of zein when low molecular weight 0.1% chitosan is added.
Figure 3 is a graph showing the particle size and zeta potential of the complex according to the concentration of zein when low molecular weight 0.5% chitosan is added.
Figure 4 is a graph showing the particle size and zeta potential of the complex according to the concentration of zein when low molecular weight 1% chitosan is added.
5 is a graph showing the particle size and zeta potential of the composite according to the concentration of zein when high molecular weight 0.5% chitosan is added.
6 is a graph showing the particle size and zeta potential of the complex according to the concentration of zein when high molecular weight 1% chitosan is added.
7 is a graph showing the particle size and zeta potential of the complex according to the low molecular weight chitosan concentration.
8 is a result of the release of the zein-chitosan complex according to the chitosan concentration (Zane 20mg / ml)
to be.
9 is a result of the release of the zein-chitosan complex according to the concentration of zein (chitosan 0.1%).

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the following examples

Example

Example  One. Stigmasterol  Preparation of Zane-chitosan Complexes Containing

Zein, a type of protein, was dissolved in 75% ethanol, chitosan was dissolved in 1% lactic acid, and stigmasterol was dissolved in methanol to prepare respective solutions (where to buy stigmasterol, zein and chitosan). Sigma).

The stigmasterol solution obtained by the above method was dropped into the zein solution and stirred at a speed of 600 rpm for 1 hour to coat stigmasterol with zein (coating material). Herein, the chitosan solution obtained by the above method was encapsulated by dropping at a ratio of 1:20 (chitosan solution: coating material) at a speed of 600 rpm for 1 hour to prepare a zein-chitosan complex containing stigmasterol.

Dropping 1% of Tween20 in the complex while stirring at a speed of 600rpm to adjust the concentration of Tween20 to 0.44% and ethanol was removed using a concentrator to measure particle size, zeta potential, and freeze-drying.

In the present invention, the zein may be used in place of gliadin, hordein, saclin, or kafirin.

Example  2. The Jane-chitosan Complex Stigmasterol  Confirm

The following conditions were used to analyze stigmasterol. The complex of Example 1 was filtered through a 0.45 μm membrane filter and analyzed by HPLC (HITACHI, High-Technologies Corporation, Tokyo, Japan). The column was a C ₁₈ reversed phase column (3.9 × 300 mm, Waters, USA) and the temperature of the column oven was analyzed at 30 ℃. Solvent conditions include acetonitrile containing 0.1% acetic acid and water containing 0.1% acetic acid as A and B solvents, respectively. The concentration of solvent A is increased from the initial 0% to 100% for 30 minutes. And analyzed. The flow rate of the solvent was 1 ml / min and the stigmasterol was confirmed at 254 nm using a UV detector.

Example  3. Stigmasterol  Particle size of the containing complex and Zeta potential  Measure

In the manufacturing method of Example 1, the composite was prepared under different conditions, and the particle size and the genta potential were measured accordingly.

(1) particle size measurement

The particle size of the emulsion droplets was measured using a particle size analyzer (Microtrac nanotrac 250, Microtrac Inc., USA).

(2) Zeta potential  Measure

Zeta potential is one of the important factors indicative of stability. The higher the value of the zeta potential, the higher the stability. Zeta potential was measured using a zeta potential analyzer (Zetaplus, Brookhaven Instruments corporation, USA).

3-1. Particle size of Jane nanoparticles and Zeta potential  Measure

Nanoparticles were prepared by the method of Example 1 using only zein without adding chitosan, and the particle size and the zeta potential were measured, and the particle size was very low (FIG. 1).

In addition, nanoparticles were prepared by the method of Example 1 using only chitosan without adding zein as a control, but the particle size and zeta potential were measured, but no complex was formed using only chitosan.

3-2. Particle size according to the amount of chitosan added Zeta potential  Measure

Low molecular weight (20-300cp) chitosan was added to 0.1, 0.5 and 1% of the total weight of the composite to determine the particle size and zeta potential according to the concentration of zein, respectively. As a result, the particle size also increased as the amount of chitosan increased. In the case of zeta potential, chitosan decreased as the concentration of zein increased with 0.1 and 0.5% of the total weight, and slightly increased as the concentration of zein increased with 1% but did not affect the particle size. (Figures 2-4).

On the other hand, when high molecular weight (800-2000cp) chitosan was used, the particle size and zeta potential were measured according to the concentration of zein by adding 0.5 and 1% of the total weight of the composite, respectively, and compared with the case of adding low molecular weight chitosan. It was confirmed that the particle size increases with increasing the concentration of (Fig. 5 and 6).

In addition, as a result of measuring the particle size and zeta potential according to the low molecular weight chitosan concentration, when the chitosan concentration was 0.1%, the particle size was very low (representing a stable zeta potential value) and the particle size increased as the concentration increased more than 0.5%. As a result, the zeta potential value was also lowered (FIG. 7).

Example  4. Depending on the concentration of chitosan Collection efficiency  And elution amount

A certain amount of capsules collected with stigmasterol was put into the elution medium, and sampled over time using a dissolution test apparatus (vision elite 8, Hanson, USA), followed by HPLC (Jasco, Tokyo, Japan). Measured. Elution medium was HCl solution (pH 1.2), PBS buffer (pH 7.4) was used, the temperature was 37 ℃, the stirring rate was maintained at 100rpm.

The result of changing the concentration of chitosan at 20 mg / ml and changing the concentration of chitosan is shown in FIG. 8. That is, when the concentration of chitosan was increased to 0.5%, the inhibition of elution was large. I could confirm it.

In addition, Table 1 below shows the collection efficiency ((composite content / total amount) X100) of the zein-chitosan composite prepared by varying the chitosan concentration.

Collection Efficiency of Zein-Chitosan Complex According to Chitosan Concentration (Zane 20mg / ml) Chitosan concentration (w / v) Collection efficiency (%) 0.1% 63.08 0.5% 90.43 1.0% 91.35

That is, when the chitosan concentration is increased to 0.5% compared to 0.1%, it can be seen that the efficiency is significantly increased.

Example  5. Jane's  Depending on concentration Collection efficiency  And elution amount

The experiment was conducted in the same manner as in Example 4, but the concentration of chitosan was fixed at 0.1%, and the result of changing the concentration of zein is shown in FIG. 9. As a result, it can be confirmed that dissolution inhibition appears as the concentration is higher.

In addition, the collection efficiency of the composite prepared by varying the concentration of zein in Table 2 was measured.

The collection efficiency of the zein-chitosan complex according to the concentration of zein (chitosan concentration 0.1% w / v) Jane concentration Collection efficiency (%) 10mg / ml 38.92 20mg / ml 63.08

In other words, the higher the concentration of zein was confirmed that the collection efficiency increased. This means that as the zein concentration increases, the coating efficiency increases. However, when the concentration of zein exceeds 20mg / ml did not show a significant increase in efficiency as in the above range.

While the present invention has been described with reference to exemplary embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. You will know. In addition, many modifications may be made to adapt a particular situation and material to the teachings of the invention without departing from the essential scope thereof. Accordingly, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention be construed as including all embodiments falling within the scope of the appended claims.

Claims (18)

A phytosterol-containing prolamine-cation polymer complex comprising phytosterol, prolamine and cationic polymer, wherein the prolamin and cationic polymer coats phytosterol. The method of claim 1,
The phytosterol is any one or more selected from the group consisting of β-sitosterol (β-sitosterol), stigmasterol (stigmasterol), campesterol (campesterol), sitostanol (campestanol) and campestanol (campestanol) Lolamine-Cation Polymer Composite. 3. The method of claim 2,
The phytosterol is stigmasterol (stigmasterol), characterized in that the prolamin-cation polymer composite. The method of claim 1,
The prolamin is any one or more selected from the group consisting of zein, gliadin, hordein, secalin, and kafirin. Cationic polymer composite. 5. The method of claim 4,
Prolamin-cation polymer composite, characterized in that the prolamin is zein. The method of claim 1,
And the cationic polymer is chitosan. The method according to claim 6,
The chitosan is poly [ß- (1-4) -2-amino-2-dioxy-D-glucopyranose], thiolated chitosan, trimethylated chitosan, carboxymethyl chitosan and N- (2-hydroxypropyl-). Prolamin-cationic polymer composite, characterized in that any one or more selected from the group consisting of 3-trimethyl ammonium chitosan chloride. The method of claim 1,
Prolamin-cationic polymer composite, characterized in that the molecular weight of the cationic polymer is 800-2000cp. The method of claim 1,
The concentration of the cationic polymer is 0.3 to 0.7% w / v, the concentration of zein is a prolamin-cationic polymer composite, characterized in that 15mg / ml to 25mg / ml. In mixing phytosterol, prolamine and cationic polymer,
The prolamin and cationic polymer is a method for producing a phytosterol-containing prolamin-cation polymer composite, characterized in that the coating film phytosterol (phytosterol). The method of claim 10,
The phytosterol is any one or more selected from the group consisting of β-sitosterol (β-sitosterol), stigmasterol (stigmasterol), campesterol (campesterol), sitostanol (campestanol) and campestanol (campestanol) Method for preparing a rollamine-cationic polymer composite. 12. The method of claim 11,
The phytosterol is stigmasterol (stigmasterol), characterized in that the production method for the prolamin-cation polymer composite. The method of claim 10,
The prolamin is any one or more selected from the group consisting of zein, gliadin, hordein, secalin, and kafirin. -Preparation of Cationic Polymer Composite. 14. The method of claim 13,
The prolamin is zein (zein) characterized in that the method for producing a prolamin-cation polymer composite. The method of claim 10,
The cationic polymer is chitosan, characterized in that the method for producing a prolamin-cationic polymer composite. 16. The method of claim 15,
The chitosan is poly [ß- (1-4) -2-amino-2-dioxy-D-glucopyranose], thiolated chitosan, trimethylated chitosan, carboxymethyl chitosan and N- (2-hydroxypropyl-). A method for producing a prolamin-cationic polymer composite, characterized in that any one or more selected from the group consisting of 3-trimethyl ammonium chitosan chloride. The method of claim 10,
The molecular weight of the cationic polymer is 800-2000cp, characterized in that the method for producing a prolamin-cationic polymer composite. The method of claim 10,
The concentration of the cationic polymer is 0.3 to 0.7% w / v, or the concentration of zein is 15 mg / ml to 25 mg / ml method for producing a prolamin-cationic polymer composite.

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