KR20100000917A - Complex of antioxidant and chitosan having improved stability - Google Patents

Abstract

PURPOSE: An antioxidant-chitosan complex is provided to enhance stability of the antioxidant in aqueous media and minimize denaturation of the antioxidant by temperature, light ray, oxygen, and water. CONSTITUTION: An antioxidant-chitosan complex is produced by reacting chitosan with α-lipoic acid or glutathione antioxidant and chitosan under the presence of 3-dimethyl-aminopropyl-N-ethylcarbodimide. The reaction is performed under the presence of N-hydroxysucciinimide and acid as catalyst. A method for producing the antioxidant-chitosan complex comprises: a step of reacting the antioxidant and succinic acid under the presence of 3-dimethyl-aminopropyl-N-ethyl carbodimide; and a step of reacting the 3-antioxidant-carbonyl propanoic acid and chitosan under the presence of 3-dimethyl-aminopropyl-N-ethyl carbodimide. The antioxidant is coenzyme Q10, α-tocopherol, retinol, ascorbic acid, or butylated hydroxy toluene.

Description

Complex of antioxidant and chitosan having improved stability

The present invention relates to a complex of an antioxidant and chitosan, and more particularly, to a complex prepared by directly coupling an antioxidant to chitosan or by linking a linker such as succinic acid. The zeo-chitosan complex not only effectively increases the stability of the antioxidant, but can also block the odor of the antioxidant, and thus can be usefully applied to the pharmaceutical composition and the cosmetic composition.

Pharmaceutical compositions and / or cosmetic compositions include various antioxidants. The antioxidants include α-lipoic acid, coenzyme Q10, α-tocopherol, retinol, glutathione, ascorbic acid, butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin gallate Various compounds are known, such as late, gallocatechin gallate, silybin, diosmethin, camphorol, epicatechin, galangin and the like.

For example, glutathione plays an important role in the respiration of mammals and plant tissues, protects red blood cells by reducing hydrogen peroxide, a harmful by-product that occurs during various metabolic reactions, into water, and acts as a cofactor for various enzymes to regenerate immune cells. To get involved. In the elderly, 75 mg of glutathione per day has been reported to significantly increase the activity of immune cells. Coenzyme Q10 is a coenzyme that helps mitochondria create energy. It has been reported to have various pharmacological actions including antiviral, antibacterial and anticancer activity. In addition, tocopherol helps increase oxygen in blood, improves blood circulation, prevents arteriosclerosis, maintains elasticity of blood vessels and prevents blood coagulation, and has been reported to prevent and treat diseases such as cardiovascular diseases. .

On the other hand, when the antioxidants are present alone, there is a problem that the stability in a special aqueous medium is low, such as easily denatured (for example, reduced), depending on the antioxidants, the generation of inverse odor, that is, the problem of smell.

For example, α-lipoic acid, which is known to have various pharmacological effects such as enhancing the body's immune function, lowering blood sugar, and suppressing appetite, is easily reduced to produce dihydrolipoic acid, thereby causing an adverse smell. There is a problem that occurs. In order to solve this problem, encapsulation using liposomes has been proposed, but it is very difficult to encapsulate an excess of α-lipoic acid.

In addition, ascorbic acid, which is known as a representative antioxidant, has a structure similar to Y-lactone, and thus has a problem of being easily decomposed by reacting sensitively to external environments such as air, especially oxygen, heat, and light. As a solution to the problem of ascorbic acid stability, oxidation of ascorbic acid can be achieved by adding antioxidants, stabilizing in multiple emulsions, stabilizing in oil-based emulsions, and using zinc sulfate and L-tyrosine together. Inhibitors have been proposed (US Pat. No. 4,938,969, EP 553,667 B1, etc.). In addition, to improve the stability of ascorbic acid, sodium ascorbylphosphate, sodium ascorbyl phosphate, magnesium ascorbylphosphate, calcium ascorbylphosphate, and ascorbic acid polypeptide , Ethyl ascorbyl ether, ascorbyl dipalmitate, ascorbyl palmitate, ascorbyl glucoside, ascorbyl glucoside, ascorbyl ethylsilanol pectinate Examples of modifications with derivatives such as Ascorbyl ethylsilanol pectinate have been reported.

The inventors have conducted various studies to develop a method that can increase the stability of various antioxidants, including α-lipoic acid, glutathione, ascorbic acid and the like, especially in aqueous media. As a result, when the antioxidant was prepared in complex form with chitosan, it was found that the obtained antioxidant-chitosan complex not only has excellent stability but also can fundamentally block the problem of the deodorant of the antioxidant. In addition, since the antioxidant-chitosan complex may be present as an antioxidant and chitosan by decomposing the antioxidant-chitosan complex in a living environment (for example, a gastrointestinal acid having an acidic pH) or in skin tissue, the antioxidant-chitosan complex may be elevated. It has been found that synergistic effects can be expected in the pharmacological effects such as synergy effects, for example, whitening, anti-aging, skin protection, wrinkle improvement, and skin moisturizing.

Accordingly, the present invention aims to provide a complex of an antioxidant and chitosan.

According to an aspect of the present invention, an antioxidant selected from the group consisting of α-lipoic acid and glutathione and chitosan are obtained by a process comprising the step of reacting in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. Antioxidant-chitosan complexes are provided.

The reaction may preferably be carried out in the presence of N-hydroxysuccinimide and an acid as a catalyst, the acid including acetic acid, sulfonic acid, sulfuric acid, phosphoric acid and the like. Further, after performing the reaction, the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably molecular weight cut-off of about 2 000, followed by drying. It may further comprise.

According to another aspect of the invention, (a) coenzyme Q10, α-tocopherol, retinol, ascorbic acid, butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin Reaction of succinic acid with at least one antioxidant selected from the group consisting of gallate, gallocatechin gallate, silibine, diosmethine, camphorol, epicatechin, and galangin in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide To prepare 3-antioxidant-carbonyl propanoic acid; And (b) reacting the 3-antioxidant-carbonyl propanoic acid and chitosan obtained in step (a) in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. A chitosan complex is provided.

More preferably, the antioxidant is retinol, coenzyme Q10, α-tocopherol, or ascorbic acid. The reaction of step (a) may preferably be carried out in the presence of a base selected from the group consisting of dimethylaminopyridine, triethylamine, and isopropylethylamine. The reaction of step (b) may preferably be carried out in the presence of N-hydroxysuccinimide and an acid as a catalyst, the acid comprising acetic acid, sulfonic acid, sulfuric acid, phosphoric acid and the like. Further, after carrying out the reaction of step (b), the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably molecular weight cut-off of about 2 000. It may further comprise the step of drying.

The antioxidant-chitosan complex according to the present invention effectively increases the stability of the antioxidant in the aqueous medium, thereby minimizing the alteration caused by the environment such as temperature, light, oxygen and water even if stored in the aqueous composition for a long time. In particular, when an antioxidant such as α-lipoic acid is prepared in a complex with chitosan according to the present invention, it is possible to fundamentally block the occurrence of inverse odor due to degeneration (eg, reduction) of the antioxidant, that is, the problem of malodor. . In addition, chitosan is a non-toxic, biodegradable, bio-compatible material, in which the antioxidant-chitosan complexes are degraded and present as antioxidants and chitosan in vivo environment (e.g., gastrointestinal tract with acidic pH) or skin tissue. As a result, synergistic effects with antioxidants, for example, whitening, anti-aging, skin protection, anti-wrinkle, skin moisturizing effect can be expected synergistic effect. Therefore, the antioxidant-chitosan complex according to the present invention can be usefully applied to pharmaceutical compositions and cosmetic compositions.

The present invention provides an antioxidant-chitosan complex obtained by a process comprising reacting an antioxidant selected from the group consisting of α-lipoic acid and glutathione with chitosan in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. To provide.

The chitosan used in the preparation of the antioxidant-chitosan complex of the present invention is a polymer obtained by deacetylating chitin (see Structural Formula of Formula 1) in which an N-acetyl-glucosamine unit is oxygen-linked (see Structural Formula of Formula 2). Biodegradable and bio-compatible.

Chitosan may exist in various forms according to the degree of deacetylation and / or the degree of polymerization of the glucosamine unit, and the degree of deacetylation and weight average molecular weight of the chitosan used in the preparation of the antioxidant-chitosan complex according to the present invention is large. It is not limited. Preferably, the degree of deacetylation of chitosan may be in the range of 40 to 90%, more preferably 70 to 90%, and the weight average molecular weight may be 200 kDa to 600 kDa, more preferably about 300 kDa.

Chitosan not only inhibits the absorption of bile acids in the intestine, but also lowers the concentration of bile acids and dissolves blood clots to prevent atherosclerosis, heart disease, stroke and sudden death. It is known to be useful for diet because it absorbs excess fat in the body and discharges it out of the body. In addition, it is known to absorb chlorine in the intestine and discharge it to the outside to prevent the occurrence of high blood pressure, promote the growth of enteric beneficial bacteria, inhibit the growth of harmful bacteria to promote the absorption of nutrients and improve digestive function. In addition, chitosan is known to have an anticancer effect, increase the resistance of immune cells, and prevent adult diseases by adsorption of harmful substances in the large intestine and blood vessels.

The reaction of the antioxidant with chitosan is carried out using a coupling agent, i.e. 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The antioxidant can be added by dissolving in an organic solvent such as tetrahydrofuran, dimethylformamide and the like. The coupling reaction may be preferably carried out in the presence of N-hydroxysuccinimide and an acid as a catalyst, and the acid may include acetic acid, sulfonic acid, sulfuric acid, phosphoric acid, and the like, preferably acetic acid. In addition, the coupling reaction may be performed at a temperature of 20 ° C. to about 50 ° C., more preferably at room temperature (about 25 ° C.) to prevent antioxidants such as α-lipoic acid from forming polymers.

Although the equivalent ratio of the antioxidant and chitosan may vary depending on the degree of deacetylation of chitosan, it is unreacted to use 0.5 to 1.5 equivalents, more preferably 0.9 to 1 equivalents, of antioxidant per 1 equivalent of chitosan. May not leave an antioxidant (for example, α-lipoic acid), thereby facilitating purification.

As described above, the antioxidant-chitosan complex obtained by the coupling reaction has auto-emulsifying properties by having water solubility and fat solubility at the same time, and therefore, separation by dialysis using a semipermeable membrane is preferable. That is, after performing the coupling reaction, the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably molecular weight cut-off of about 2 000, thereby removing the unreacted material. After removal, it is preferable to separate the antioxidant-chitosan complex by drying the obtained filtrate. The drying may be carried out by ordinary vacuum drying, lyophilization and the like, more preferably by lyophilization.

The antioxidant-chitosan complex of the present invention can be obtained by reacting an antioxidant with chitosan via a linker. That is, the present invention includes, in addition to α-lipoic acid and glutathione, an antioxidant-chitosan complex obtained by linking various antioxidants with chitosan via a linker having a carboxylic acid moiety on both sides, preferably succinic acid. Specifically, the present invention comprises the steps of (a) reacting the antioxidant and succinic acid in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide to prepare 3-antioxidant-carbonyl propanoic acid; And (b) reacting the 3-antioxidant-carbonyl propanoic acid and chitosan obtained in step (a) in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. A chitosan complex.

Antioxidants that can be applied to the linker-mediated complex formation include various antioxidants, for example, coenzyme Q10, α-tocopherol, retinol, ascorbic acid, Butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin gallate, gallocatechin gallate, It may be selected from the group consisting of silbin, diosmetin, kaempferol, epicatechin, and galangin. More preferably, the antioxidant may be retinol, coenzyme Q10, α-tocopherol, or ascorbic acid.

The reaction of the antioxidant of succinic acid in step (a) is carried out using a coupling agent, i.e. 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The reaction can be carried out in an organic solvent such as dichloromethane, tetrahydrofuran, chloroform, ethyl acetate and the like. In addition, the reaction may be preferably carried out in the presence of a base selected from the group consisting of dimethylaminopyridine, triethylamine, and isopropylethylamine. The equivalent ratio of the antioxidant and succinic acid may be in the range of 0.5 to 0.6 equivalents, more preferably about 0.5 equivalents, based on 1 equivalent of antioxidant. The 3-antioxidant-carbonyl propanoic acid obtained from the above reaction can be separated by distillation or drying of the reaction mixture under reduced pressure. The reaction mixture is concentrated under reduced pressure, if necessary, and then a solvent such as ethyl acetate is added, followed by aqueous hydrochloric acid solution. After washing one or more times can be separated by drying.

The reaction of the 3-antioxidant-carbonyl propanoic acid with chitosan in step (b) is carried out using a coupling agent, ie 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The coupling reaction may be preferably carried out in the presence of N-hydroxysuccinimide and an acid as a catalyst, the acid may include acetic acid, sulfonic acid, sulfuric acid, phosphoric acid and the like, preferably acetic acid may be used. . In addition, the coupling reaction can be performed at 20 ° C. to about 50 ° C., more preferably at room temperature (about 25 ° C.) to prevent the 3-antioxidant-carbonyl propanoic acid from forming polymers. Although the equivalent ratio of the 3-antioxidant-carbonyl propanoic acid and chitosan may vary depending on the degree of deacetylation of chitosan, the amount of 3-antioxidant-carbonyl propanoic acid is 0.9-1.5 equivalents to 1 equivalent of chitosan, More preferably, the use in the range of 0.9 to 1 equivalent may not leave unreacted 3-antioxidant-carbonyl propanoic acid, so that purification is easy.

Since the antioxidant-chitosan complex (specifically, the antioxidant-linker-chitosan complex) obtained as described above has water-soluble and fat-soluble properties at the same time, it has an auto-emulsifying property, so that separation by dialysis using a semipermeable membrane is recommended. desirable. That is, after performing the coupling reaction, the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably molecular weight cut-off of about 2 000, thereby removing the unreacted material. After removal, it is preferable to separate the antioxidant-chitosan complex by drying the obtained filtrate. The drying may be carried out by ordinary vacuum drying, lyophilization and the like, more preferably by lyophilization.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are intended to illustrate the invention and should not be construed as limiting the invention.

Example  1: Complex synthesis of α-lipoic acid and chitosan

5.00 g (12.75 mmol) of chitosan at 90% deacetylation degree and 300 kDa of weight average molecular weight, 2.90 g (15.30 mmol) of 3-dimethyl-aminopropyl-N-ethyl carbodiimide, and 1.76 g of N-hydroxysuccinimide (15.30 mmol) was dissolved in 250 ml of a 1% acetic acid aqueous solution. To the obtained solution, a solution in which 3.16 g (15.30 mmol) of α-lipoic acid was dissolved in 30 ml of tetrahydrofuran was added, followed by reacting at room temperature for 72 hours. 500 ml of tetrahydrofuran was added to the reaction mixture, and the precipitate was separated by filtration. The obtained precipitate was completely dissolved in 300 ml of water, and then the Milipore dialysis tube, M.W. Unreacted material was removed by semipermeable membrane filtration for 2 days with cut-off 2,000. The obtained filtrate was lyophilized to obtain 6.14 g (yield 83%) of a complex of α-lipoic acid and chitosan.

¹ H NMR 400 MHz ppm 2.51 to 2.61 (α-lipoic acid, -SCH-, -SCH _2- , 3H, m) 1.55 to 2.18 (α-lipoic acid, -CH _2- , 8H, m); FT-IR Cm ^-1 3393.13 (-OH, br) 2928.29 (-CH _2- , w) 1645.82 (-NHCO-, s) 1553.28 (-NHCO-, s)

Example  2 : Glutathione  Complex synthesis of chitosan

The reaction was carried out in the same manner as in Example 1, except that 4.70 g (15.30 mmol) of glutathione was used instead of α-lipoic acid to obtain 5.94 g (yield 76%) of the complex of glutathione and chitosan.

¹ H NMR 400 MHz ppm 4.09 (glutathione, -NHCO CH ₂ NH-, 2H, t) 3.49 (glutathione, -CH ₂ CH ₂ CHNH _2- , 2 H, t) 3.18-2.93 (glutathione, -CH CH ₂ SH, 2H, m) 2.18 (gluthathione, -CO CH ₂ CH _2- , 2H, t); FT-IR Cm ^-1 3392.73 (-OH, br) 2927.32 (-CH _2- , w) 1682.26, 1679.64, 1644.64, 1552.15, (-NHCO-, s)

Example  3: 3-retinol-carbonyl Propanoic acid  synthesis

5.00 g (17.45 mmol) of retinol and 2.47 g (20.95 mmol) of succinic acid were added to 70 ml of dichloromethane with stirring, and 40.16 g (20.95 mmol) of 3-dimethyl-aminopropyl-N-ethyl carbodiimide was added over 30 minutes. Slowly added. 0.43 g (3.19 mmol) of dimethylaminopyridine was added to the reaction mixture, which was then stirred at room temperature for 9 hours. The reaction mixture was concentrated under reduced pressure, and then 100 ml of ethyl acetate was added. The reaction mixture was washed twice with 1N aqueous hydrochloric acid solution, concentrated under reduced pressure to remove the solvent, and then dried in vacuo to give 5.46 g (yield 81%) of 3-retinol-carbonyl propanoic acid as a solid.

¹ H NMR 400 MHz ppm 6.51-5.63 (retionol, -C = CH- , 6H, s) 4.75 (retinol, -OCH _2- , 2H, d) 2.62-2.52 (succinic, -CH ₂ CH _2- , 4H, m ) 1.96-1.57 (retinol ring, -CH _2- , 6H, m) 1.21 (retinol ring, -CH ₃ , 6H, s); FT-IR C m ^-1 3384.16 (-OH, br) 2982.34-2931.97 (-CH _2- , w) 1714.67 ( -CO 0-, s) 1516.27 (-C = CH-, m)

Example 4 Synthesis of 3-Reduced Coenzyme Q10-Carbonyl Propanoic Acid

Reduced instead of retinol The reaction was carried out in the same manner as in Example 3, except that 5.00 g (12.80 mmol) of coenzyme Q10 was used to obtain 6.93 g (yield 78%) of 3-reduced coenzyme Q10-carbonyl propanoic acid.

¹ H NMR 400 MHz ppm 5.80-5.20 (coenzyme Q ₁₀ , -C = CH- , 3H, s) 3.73 (coenzyme Q ₁₀ , -OCH _3- , 6H, s) 2.54-2.51 (succinic, -CH ₂ CH _2- , 4H, m) 2.12-3.22 (coenzyme Q ₁₀ , -CH _2- , 10H, m) 2.35 (coenzyme Q ₁₀ , -CH ₃ , 3H, s) 1.73-1.69 (coenzyme Q ₁₀ , -CH ₃ , 12H, s); FT-IR C m ^-1 3411.27 (-OH, br) 2979.36-2947.56 (-CH _2- , w) 1716.32 ( -CO 0-, s) 1516.71 (-C = CH-, m)

Example  5: 3-tocopherol-carbonyl Propanoic acid  synthesis

The reaction was carried out in the same manner as in Example 3, except that 5.00 g (17.46 mmol) of α-tocopherol was used instead of retinol to obtain 7.97 g (yield 86%) of 3-tocopherol-carbonyl propanoic acid.

¹ H NMR 400 MHz ppm 2.63-2.09 (tocopherol ring, -CH ₂ CH _2- , 4H, m) 2.56-2.48 (succinic, -CH _2- , 4H, m) 2.35 (tocopherol, -CH ₃ , 9H, s) 1.83 (tocopherol, 1H, s) 1.65-1.25 (tocopherol, -CH _2- , 18H, m) 1.01-1.06 (tocopherol, -CH ₃ , 15H, s); FT-IR C m ^-1 3413.18 (-OH, br) 2978.65-2932.49 (-CH _2- , w) 1716.87 ( -CO 0-, s) 1516.83 (-C = CH-, m)

Example  6: 3-ascorbic acid-carbonyl Propanoic acid  synthesis

The reaction was carried out in the same manner as in Example 3, except that 5.00 g (28.39 mmol) of ascorbic acid was used instead of retinol to give 5.72 g (yield 73%) of 3-ascorbic acid-carbonyl propanoic acid.

^{1 H NMR 400MHz ppm 4.71 (ascorbic} ring, - CH-, 1H, s) 3.94 (ascrobic acid, -CH-, 1H, m) 3.78 (ascorbic acid, -CH 2 -, 2H, m) 2.54 - 2.45 (succinic , -CH _2- , 4H, m); FT-IR C m ^-1 3392.27 (-OH, br) 2983.47-2987.68 (-CH _2- , w) 1716.87 ( -CO 0-, s) 1514.59 (-C = CH-, m)

Example  7: Retinol and  Synthesis of Complexes of Chitosan

The reaction was carried out in the same manner as in Example 1, except that 5.91 g (15.30 mmol) of 3-retinol-carbonyl propanoic acid obtained in Example 3 was used instead of α-lipoic acid, thereby preparing a complex of retinol and chitosan ( That is, 8.83 g (yield 73%) of retinol-linker-chitosan complex) were obtained.

¹ H NMR 400 MHz ppm 6.49-5.62 (retionol, -C = CH- , 6H, s) 4.69 (retinol, -OCH _2- , 2H, s) 2.62-2.52 (succinic, -CH ₂ CH _2- , 4H, m ) 1.89-1.52 (retinol ring, -CH _2- , 6H, m) 1.22 (retinol ring, -CH ₃ , 6H, s); FT-IR C m ^-1 3346.34 (-OH, br) 2956.85-2936. 75 (-CH _2- , w) 1718.49 ( -CO O-, s) 1644.58 (-NHCO-, s) 1553.93 (-NHCO-, s ) 1516.71 (-C = CH-, m)

Example  8: reduced coenzyme Q10 And synthesis of chitosan complex

The reaction was carried out in the same manner as in Example 1, except that 7.48 g (15.30 mmol) of 3-coenzyme Q10-carbonyl propanoic acid obtained in Example 4 was used instead of α-lipoic acid to reduce the reduced coenzyme Q10 and chitosan. 7.32 g (yield 69%) of the complex (ie, the reduced coenzyme Q10-linker-chitosan complex) was obtained.

¹ H NMR 400 MHz ppm 5.24 (coenzyme Q ₁₀ , -C = CH- , 3H, s) 3.50 (coenzyme Q ₁₀ , -OCH ₃ , 6H, s) 2.57-2.53 (succinic, -CH ₂ CH _2- , 4H, m) 2.12-2.63 (coenzyme Q ₁₀ , -CH _2- , 10H, m) 1.92 (coenzyme Q ₁₀ ring, -CH ₃ , 3H) 1.71-1.74 (coenzyme Q ₁₀ , -CH ₃ , 15H, s); FT-IR C m ^-1 3393.48 (-OH, br) 2929.46 (-CH _2- , w) 1716.68 ( -CO O-, s) 1646.75 (-NHCO-, s) 1554.65 (-NHCO-, s) 1517.74 (- C = CH-, m)

Example  9: Synthesis of Complex of Tocopherol and Chitosan

The reaction was carried out in the same manner as in Example 1, except that 5.91 g (15.30 mmol) of 3-tocopherol-carbonyl propanoic acid obtained in Example 5 was used instead of α-lipoic acid, and thus a complex of tocopherol and chitosan (ie, 8.83 g (yield 73) of tocopherol-linker-chitosan complex) were obtained.

¹ H NMR 400 MHz ppm 2.62-2.06 (tocopherol ring, -CH ₂ CH _2- , 4H, m) 2.52-2.43 (succinic, -CH _2- , 4H, m) 2.34 (tocopherol ring, -CH ₃ , 9H, s ) 1.79 (tocopherol, 1H, s) 1.63-1.28 (tocopherol, -CH _2- , 18H, m) 1.52-1.03 (tocopherol, -CH ₃ , 15H, s); FT-IR C m ^-1 3362.72 (-OH, br) 2994.73-2924.69 (-CH _2- , w) 1722.71 ( -CO O-, s) 1643.37 (-NHCO-, s) 1552.81 (-NHCO-, s ) 1425.78 (-C = C-, m)

Example  10: complex synthesis of ascorbic acid and chitosan

The reaction was carried out in the same manner as in Example 1, except that 4.23 g (15.30 mmol) of 3-ascorbic acid-carbonyl propanoic acid obtained in Example 6 was used instead of α-lipoic acid, and a complex of ascorbic acid and chitosan (ie, 6.59 g (yield 71%) of ascorbic acid-linker-chitosan complex) were obtained.

^{1 H NMR 400MHz ppm 1 H NMR} 400MHz ppm 4.71 (ascorbic ring, - CH -, 1H, s) 3.94 (ascrobic acid, -CH-, 1H, m) 3.76 (ascorbic acid, -CH 2 -, 2H, m) 2.49-2.41 (succinic, -CH _2- , 4H, m); FT-IR C m ^-1 3383.64 (-OH, br) 2997.81-2913. 54 (-CH _2- , w) 1722.39 ( -CO O-, s) 1641.48 (-NHCO-, s) 1548.72 (-NHCO-, s ) 1422.36 (-C = C-, m)

Test example. Properties, chemical stability, and sensory tests

3.0 g of each complex of the antioxidants and chitosan prepared in Examples 1, 2, 7, 8, 9, and 10 were completely dissolved in 1,000 ml of distilled water to prepare a clear solution, and stirred at 40 ° C. for 30 minutes, and micelle (micelle) aqueous solution was prepared. 10 ml of the obtained aqueous solution was taken and stored at 45 ° C. for 0 weeks, 1 week, 2 weeks, 3 weeks, and 4 weeks, whereby a change in properties, a residual content of an antioxidant, and a sensory test were performed.

The change in appearance was visually observed.

The residual content of the antioxidant was inverted by measuring the content (measurement content) of the antioxidant eluted from the antioxidant-chitosan complex into the aqueous solution by high performance liquid chromatography. In the high performance liquid chromatography, each sample was taken in a 10 ml-volume flask, each 0.1 g of the sample, adjusted to 10 ml with chloroform, vortexed for 1 minute, and centrifuged at 10000 rpm for 10 minutes. It was measured by high performance liquid chromatography. The high performance liquid chromatography conditions are as follows: column-ACE 5-C18 (4.6 * 150mm, 5㎛), Guard Column-ACE 5-C18 (2.1 * 12.5mm, 5㎛), mobile phase Mixed solution of acetonitrile and 0.1% aqueous sulfuric acid solution (80:20), detector wavelength-UV 224 nm, flow rate-1 ml / min, injection amount-2 μl.

Sensory tests were performed as follows: Each sample was measured and averaged by measuring the intensity of odor of 10 women in their late 20s, the intensity of odor was set as shown in Table 1 below.

Bad smell Explanation 0 Odorless One Detection 2 Normal odor 3 Strong odor 4 Extreme odor 5 Unbearable odor

As described above, the results of the property change, the residual content of the antioxidant, and the sensory test are shown in Table 2 below.

Composite (Example) Retention period Change of appearance Antioxidant Content (%) Sensory test (direct sensory method) Average value measured twice (%) Measurement content (%) Residual content (%) Example 1 Week 0 none 0.0012 0.79 99.21 1.3 1 week none 0.0017 1.12 98.88 1.6 2 weeks none 0.0023 1.51 98.49 2.1 3 weeks none 0.0032 2.11 97.89 2.7 4 Weeks none 0.0058 3.82 96.18 2.6 Example 2 Week 0 none 0.0022 1.24 98.76 1.3 1 week none 0.0031 1.75 98.25 1.2 2 weeks none 0.0037 2.08 97.92 1.7 3 weeks none 0.0054 3.04 96.96 2.4 4 Weeks none 0.0068 3.83 96.17 2.3 Example 7 Week 0 none 0.0041 2.14 97.86 0.7 1 week none 0.0112 5.86 94.14 0.6 2 weeks none 0.0147 7.67 92.33 1.2 3 weeks Slightly cloudy 0.0165 8.63 91.37 2.1 4 Weeks Slightly cloudy 0.0171 8.94 91.06 2.2 Example 8 Week 0 none 0.0037 1.88 98.12 0.6 1 week none 0.0066 3.35 96.65 1.7 2 weeks none 0.0118 5.99 94.01 1.5 3 weeks Slightly cloudy 0.0167 8.49 91.51 1.7 4 Weeks Slightly cloudy 0.0189 9.60 90.40 1.9 Example 9 Week 0 none 0.0034 1.96 98.04 0.7 1 week none 0.0041 2.37 97.63 0.7 2 weeks none 0.0048 2.77 97.23 1.1 3 weeks none 0.0057 3.29 96.71 1.3 4 Weeks none 0.0063 3.64 96.36 1.6 Example 10 Week 0 none 0.0031 0.55 99.45 0.8 1 week none 0.0094 1.66 98.34 0.7 2 weeks none 0.0133 2.34 97.66 1.4 3 weeks Pale yellow 0.0167 2.94 97.06 1.6 4 Weeks Pale yellow 0.0180 3.17 96.83 1.9

From the results of Table 2, when the antioxidants are easily discolored according to the present invention prepared with a complex with chitosan, it shows a high stability by maintaining a high residual content of at least 90.4% as a whole in an aqueous medium, there is little change in appearance In addition, it can be seen that there is little problem of malodor.

Claims (10)

An antioxidant-chitosan complex obtained by the production method comprising the step of reacting chitosan with an antioxidant selected from the group consisting of α-lipoic acid and glutathione in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The antioxidant-chitosan complex according to claim 1, wherein the reaction is carried out in the presence of N-hydroxysuccinimide and an acid as a catalyst. The antioxidant-chitosan complex according to claim 2, wherein the acid is acetic acid, sulfonic acid, sulfuric acid, or phosphoric acid. The process according to any one of claims 1 to 3, wherein after carrying out the reaction, the precipitate separated from the reaction mixture is dispersed in water, filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, and then drying. Antioxidant-chitosan complex, further comprising. (a) coenzyme Q10, α-tocopherol, retinol, ascorbic acid, butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin gallate, gallocatechin gallate, 3-antioxidant-carbonyl by reacting succinic acid with at least one antioxidant selected from the group consisting of silibine, diosmethine, camphorol, epicatechin, and galangin in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide Preparing propanoic acid; And (b) the antioxidant obtained by the preparation method comprising the step of reacting the 3-antioxidant-carbonyl propanoic acid and chitosan obtained in step (a) in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide- Chitosan Complex. The antioxidant-chitosan complex according to claim 5, wherein the antioxidant is retinol, coenzyme Q10, α-tocopherol, or ascorbic acid. 6. The antioxidant-chitosan complex according to claim 5, wherein said reaction of step (a) is carried out in the presence of a base selected from the group consisting of dimethylaminopyridine, triethylamine, and isopropylethylamine. The antioxidant-chitosan complex according to claim 5, wherein the reaction of step (b) is carried out in the presence of N-hydroxysuccinimide and an acid as a catalyst. The antioxidant-chitosan complex according to claim 8, wherein the acid is acetic acid, sulfonic acid, sulfuric acid, or phosphoric acid. The process according to any one of claims 5 to 9, after carrying out the reaction of step (b), the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, Antioxidant-chitosan complex, further comprising the step of drying.