KR20100087418A - Method for preparation of soluble chitosan having deacetylation of 40-60% and oral medicine using this - Google Patents

Abstract

PURPOSE: A method for simply preparing water-soluble chitosan which is deacetylated by 40-60% is provided to be safely used without side effects to human body and to obtain strong antibiotic effect, antioxidant effect, and cell activation effect. CONSTITUTION: A method for preparing water-soluble chitosan includes a step of manufacturing the water-soluble chitosan which 40-60% is deacetylated by adding acetic acid, acetic anhydride, and KOH to the chitosan which 100% is deacetylated. The amount of the acetic acid is 90-110%(v/w) of the weight of the chitosan. The amount of the KOH is the 90-110%(w/w) of the weight of the chitosan and the KOH is capable of neutralizing a chitosan solution. The amount of the acetic anhydride is 20-50%(v/w) of the weight of the chitosan.

Description

Method for preparation of soluble chitosan having deacetylation of 40-60% and oral medicine using this}

The present invention relates to a water-soluble chitosan having a deacetylation degree of 40-60%, and more particularly, to a method for preparing a water-soluble chitosan having a deacetization degree of 40-60% which can be produced with chitosan having various molecular weights and utilizing the same. Oral preparations (oral cleaning agents and oral breathing agents).

Conventional water soluble chitin. A method for producing chitosan, the chitin. Various methods have been tried for the acceptance of chitosan.

First, the molecular weight is lowered to oligosaccharides.

However, the oligosaccharides produced are natural polymer chitin, although they have many useful points due to the ease of encapsulation, water solubility, enhanced absorption of chitosan in the body, and activation of useful intestinal bacteria. There is also the adverse effect of giving up many of the good things inherent in chitosan. Among them, polymer chitin, which is the driving force that enables the continuous antibacterial activity, has a particularly low adsorption and cohesion. The coating action of the tooth surface by chitosan also cannot be expected.

Secondly, chitosan is dissolved in organic acid and dried to produce chitosan organic acid salt.

This method expands the usability of chitosan by dissolving it in acid and filtering it without using acid, filtering it, and easily encapsulating it, and easily absorbing it. However, the bitter taste of chitosan still causes many disorders and requires masking agents. In addition, this chitosan organic acid salt is stable when dissolved in water and pH is lower than 5.5. As the liquid gets more neutral, chitosan precipitates and the solution becomes turbid.

The third is chitin. It is a method of making water-soluble derivatives by polymerizing various functional groups on chitosan.

Several derivatives have already been made, and their use has diversified. For example, carboxymethyl chitin and hydroxypropyl chitin are widely used for medical and cosmetic applications, and in addition, there are continuous studies on these derivatives such as copolymers using acrylic polymers are used in wastewater and fibers. It is not endless. However, since the preparation of such derivatives is relatively difficult and expensive, it is not economical and difficult to mass-produce, and it is difficult to expect the expansion of the application range.

Accordingly, the present inventors have conducted extensive research to solve the above problems, and as a result, the inventors have invented water solubility if the degree of deacetization in the chitosan component is in the 40-60% range.

Already chitin. Chitosan has a well-established position as a food for specific health by oral administration. It has antibacterial and bactericidal effects, high affinity with human tissues, and does not cause rejection reactions. Although various developments have been made in high-tech medicines such as therapeutic agents, there is no consensus among researchers about the tissue reaction and tissue activating activity during bio-application based on the concentration, shape, degree of deacetization, molecular weight, and the like. In particular, it is necessary to examine in more detail the absorption absorption process that is important when applied directly to biological tissues.

Accordingly, the present invention provides chitin having various functions and uses. In solubilizing chitosan, the object is to provide a water-soluble chitosan with 40-60% deacetylation in its most natural form.

The present invention also provides a method for making water-soluble chitosan, which has undergone such deacetization, from 40-60% relatively easily from chitosan having various molecular weights.

The present invention is chitin. It can be used for the entire application range of chitosan (clothing, textiles, food, cosmetics, etc.) and its purpose is to supply breakthrough ingredients to health drinks, alcoholic disorders, oral preparations, chewing gum, foods, etc. There is this.

Of the above application, in particular, by preparing oral preparations having a wide pH stability, tasteless, odorless, strong adsorption power that other water-soluble chitosan does not have, can be used safely without side effects on the human body, and also to inhibit the development and growth of bacteria It is possible to achieve the original purpose of prevention and treatment of periodontal disease and gum disease caused by the oral cavity, and to effectively mask bad smell-causing substances such as methylmercaptan and garlic odor in the oral cavity and to continuously suppress and remove bad breath. It was improved to suit the symbol of.

Looking at the present invention in more detail as follows.

The water-soluble chitosan having a deacetylation degree of 40-60% of the present invention is prepared by adding acetic acid, acetic anhydride, and KOH to chitosan having a deacetylation degree of 100% having various molecular weights.

The water-soluble chitosan having undergone 40-60% deacetization according to the present invention may have various molecular weights and may have various degrees of deacetization.

According to the present invention, the amount of acetic acid is suitable for 90-110% (v / w) of the weight of chitosan, which is enough to dissolve chitosan, and the amount of KOH is 90-110% of the weight of chitosan, which is enough to neutralize the chitosan solution. (w / w) is suitable, and the most important amount of acetic anhydride is preferably 20-50% (v / w) of the weight of chitosan.

In the present invention, the degree of deacetization was most sensitive to the amount of acetic anhydride and the reaction time.

The water-soluble chitosan having undergone 40-60% deacetization according to the present invention can be used in powder form, gel form or liquid form by controlling moisture. In addition, in addition to the above components, if necessary, functional ingredients such as vitamins may be further included as other additives.

Among them, oral preparations using water-soluble chitosan having a deacetization degree of 40-60% were preferred for removing periodontal disease and calculus by inhibiting dental proteins, periodontal glycoproteins, and periodontal bacteria.

Water-soluble chitosan 40 to 60% deacetylation of the present invention can be prepared using chitosan having a variety of viscosity,

It has a pH stability and has the advantages of both chitin and chitosan.

That is, since it is water-soluble, it is water-soluble, so that it can be easily used, and is less sensitive to pH than ordinary chitosan, so there is less fear of alteration when mixed with other drugs.

In the present invention, the lower the degree of deacetylation, the higher the stability against pH, while having too low the degree of deacetylation increased the insoluble content (DA 30% or less). In addition, as the degree of deacetization (DA) was higher (over 60%), the astringent taste was gradually increased, and the stability to pH was also lowered.

Chitosan used in the present invention is a natural biopolymer material having a cationic activity, and is effective in strong antibacterial action, antioxidant activity, cell activation action, bleeding prevention, tartar removal, and tooth decay prevention. In particular, as the chitosan is water-soluble, the range of use in the industry has increased, and in each field, the value of the water-soluble chitosan is effectively used in combination with other substances.

Oral preparations using chitosan of the present invention confirmed the excellent suitability and superiority in the application to oral preparations through experiments on the antibacterial, odor removal and oral bacteria. Due to these various excellent properties, it will solve various problems in the existing oral preparation market and expand the applicability of the pharmaceutical industry and oral preparation.

Example 1

20 g of chitosan having a viscosity of 8-10 CPS and 100% deacetization is dissolved in a mixed solution of 20 ml of glacial acetic acid and 2 liters of water. 6 ml of acetic anhydride was added thereto, stirred for about 1 hour, and 12 g of KOH was dissolved in 100 ml of water. Again 2 ml of acetic anhydride are added. After stirring for 30 minutes, 6 g of KOH was added to 50 ml of water. The stirring reaction was completed for 1 hour. Measure the pH to see if it is neutralized and filter. After washing the alcohol, dried under reduced pressure at 50 ℃ DA. Check the viscosity.

Example 2

It carried out similarly to Example 1 except having increased the quantity of acetic anhydride added initially to 7 ml.

Example 3

The same procedure as in Example 1 was carried out except that chitosan having a viscosity of 80-100 CPS and a deacetization degree of 100% was used.

Example 4

It carried out similarly to Example 3 except having reduced the quantity of acetic anhydride added initially to 5 ml.

Example 5

The same procedure as in Example 1 was carried out except that chitosan having a viscosity of 800 CPS and a deacetization degree of 100% was used.

Example 6

After adding 6 ml of acetic anhydride, it carried out similarly to Example 5 except having made reaction time into 2 hours.

The analysis process was carried out as follows.

<Deacetization degree measurement>

The free amino group is measured by the clade titration method, and the degree of deacetization is determined by the following equation.

Deacetization degree (%) = free amino group X / 161

                    --------------------------- = -------------- × 100

                    Free amino group + bound amino group X / 161 + Y / 203

Where X = 1/400 × 1/1000 × f × 161 × V

        Y = 0.5 / 100-X

     V: 1 / 400N PVSK titration value (ml)

     f: factor of 1 / 400N PVSK

    161: equivalent molecular weight of the chitosan structural unit

    203: equivalent molecular weight of the structural unit of the chitin

Accurately weigh 1.0 g (as an anhydride) of chitosan, dissolve it in 0.5% (v / v) acetic acid to make exactly 200 g. The 1g is correctly taken to a 200 ml Erlenmeyer flask, 30 ml of water is added, and mixed sufficiently.

After adding 2-3 drops of 0.1% toludine blue solution as an indicator, it was titrated in 1 / 400N PVSK solution.

<Viscosity measurement>

Accurately weigh 5.0 g (as anhydride) of chitosan, dissolve it in 0.5% (v / v) acetic acid to make 500 g accurately. After stirring and dissolving and filtering, it is left to stand for 1 hour to remove the air chamber. The solution was measured using a rotational viscometer.

The deacetylation degree and viscosity measured are shown in Table 1.

(Table 1)

   division  Example 1  Example 2  Example 3  Example 4  Example 5  Example 6   DA (%)    49.1    30.8    52.7    58.2    50.4    42.1 Viscosity (CPS)   15-16   20-21    155    121    850    930

Experimental Example 1 Antibacterial Effect of Chitosan on Oral Bacteria

As the test solution, water-soluble chitosan of Example 1 of the present invention was used, and 1 mg / ml of chlorohexidine was used as a positive control solution to secure the validity of the experiment. Chlorohexidine has been widely used since the 1970s as an effective antibacterial agent for oral use.

In this experiment, Streptococcus mutans (ACTT 25175), Streptococcus sanguis (ACTT 10556), Actinomyces viscocus (ACTT 15987), Porphyro-monas gingivalis (ACTT 33277), Candidamonas albicans (ACTT 10259) , 7 strains of Eikenella corrodens (ACTT 51724) and Fusobacterium nucleatum (ACTT 27872) were cultured to determine the antimicrobial activity.

After centrifugation (5856Xg, 4 ℃, 15 minutes) of the culture medium of the bacteria, the supernatant was removed, and diluted with a new culture solution so that the culture cell viable concentration at the start of the measurement was 5 * 10 ⁵ CFL / ml. The viable cell concentration at the start point was measured by absorbance at 660 nm using a spectrophotometer, and then confirmed by the flat plate guide method. Water-soluble chitosan and chlorohexidine were used twice in stages until the minimum bacterial growth inhibitory concentration was confirmed. 2 ml of diluted chitosan, 2 ml each of chlorohexidine and distilled water and 50 ml of culture medium were doubled and cultured for 48 hours under the appropriate conditions for each strain. Absorbance was measured at 660 nm wavelength. At this time, the lowest concentration at which the absorbance was 0.05 or less was defined as the minimum bacterial growth inhibitory concentration. When the turbidity was so severe that it was difficult to measure the correct absorbance despite the correction, it was confirmed by the flat plate guide method.

(Table 2) Minimum bacterial growth inhibitory concentration and dilution factor for each strain of water-soluble chitosan and chlorohexidine

Strain CTS 50 Chlorohexidine Dilution factor mg / ml Dilution factor mg / ml  Streptococcus mutans 8 1.25 256 0.0039  Streptococcus sanguis One 10.00 128 0.0078  Actinomyces viscocus 4 2.50 256 0.0039  Porphyro-monas gingivalis 32 0.31 2,048 0.0005  Candidamonas albicans 2 5.00 64 0.0156  Eikenella corrodens 2 5.00 128 0.0078  Fusobacterium nucleatum 128 0.08 2,048 0.0005

As shown in Table 2, the minimum growth inhibitory concentration of the water-soluble chitosan of the present invention was the lowest at 0.08 mg / ml and 0.31 mg / ml for F. nucleatum and P.gingivalis, and the minimum growth rate of chlorohexidine, a positive control solution. Growth inhibitory concentration was lowest at 0.0005 mg / ml for P.gingivalis and F. nucleatum. This is a very high concentration compared to the minimum biocidal inhibitory concentration of chlorohexidine, a positive control solution, but the water-soluble chitosan of the present invention is classified as a food additive and is less restricted by the oral application concentration than chlorohexidine. Considering the fact that rollhexidine has a lot of side effects such as pigmentation on the tongue and teeth, it is highly applicable.

 Microbiological results of the present invention also showed that C. albicans is sensitive to chitosan itself. As shown in Table 2, the water-soluble chitosan of the present invention increased the antifungal activity. In other words, chitosan inhibits the adhesion of C. albicans to human oral cells and has antifungal activity.

The antimicrobial activity of the water soluble chitosan of the present invention significantly inhibited most bacterial growth tested.

Claims (3)

A process for producing a water-soluble chitosan having a deacetization degree of 40-60% by adding acetic acid, acetic anhydride, and KOH to deacetization degree 100% chitosan. The amount of acetic acid is 90-110% (v / w) of the weight of chitosan, the amount of KOH is 90-110% (w / w) of the weight of chitosan, and the amount of acetic anhydride is 20- of the weight of chitosan. A process for producing a water-soluble chitosan having a degree of deacetization of 40-60%, characterized in that 50% (v / w) is added. Oral preparation using water-soluble chitosan prepared by the method of claim 2