Abstract:
Disclosed is a stable and safe antioxidant composition which can be used routinely. Specifically disclosed is an antioxidant composition which contains one or more compounds selected from the group consisting of D-aspartic acid, derivatives and/or salts thereof. The composition may be used for the purpose of suppressing and/or improving skin conditions. The skin conditions may include, but is not limited to, fine wrinkles, a rough skin, a dry skin, a skin cancer, a skin allergy, an inflammation of the skin and a photosensitive dermatosis. The composition may be used for an external preparation for the skin, a food, and a pharmaceutical product for cataract.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a National Stage application of PCT/JP2010/055842, filed Mar. 31, 2010, which claims priority from Japanese application JP 2009-224742, filed Sep. 29, 2009. 
     TECHNICAL FIELD 
     The present invention relates to an antioxidant composition which comprises one or more compounds selected from the group consisting of D-aspartic acid, derivatives and/or salts thereof, a method of improving a skin condition comprising a step of administering the compound, and a method of treating and/or preventing cataract comprising a step of administering the compound. 
     BACKGROUND ART 
     Reactive oxygen species (ROS) oxidize non-selectively bioactive biological substances like nucleic acids, proteins, and lipids to cause damages in function of a living body or structure of an organ and a tissue. ROS are known to be a cause of skin disorders like a skin cancer, a skin allergy, an inflammation of the skin, and a photosensitive dermatosis (Non-Patent Document 1). It is also known that, with an effect on the epidermis, they cause skin conditions like fine wrinkles, a rough skin, a dry skin and the like. 
     PRIOR ART DOCUMENTS 
     Non-Patent Document 
     
         
         Non-Patent Document 1: Bickers, D. R. and Athar, M., J., Invest Dermatolg., 126:2565 (2006) 
       
    
     DISCLOSURE OF THE INVENTION 
     Problem to be Solved by the Invention 
     As an antioxidant composition, ascorbic acid (vitamin C), α(alpha)-tocopherol (vitamin E) or the like has been conventionally used for a cosmetic product and a pharmaceutical product. However, the stability is not sufficient. And therefore, there is a need to develop a stable and safe antioxidant composition that can be used routinely. 
     Means for Solving the Problem 
     In this regard, the present invention provides an antioxidant composition comprising one or more compounds selected from the group consisting of D- and/or L-aspartic acid, derivatives and/or salts thereof. 
     The antioxidant composition of the present invention may be used for suppressing and/or improving a skin condition. 
     Regarding the antioxidant composition of the present invention, the skin condition comprises, but not limited thereto, fine wrinkles, a rough skin, a dry skin, a skin cancer, a skin allergy, an inflammation of the skin, and a photosensitive dermatosis. 
     The antioxidant composition of the present invention may be used for an external preparation for the skin. 
     The antioxidant composition of the present invention may be used for a food. 
     The antioxidant composition of the present invention may be used for a pharmaceutical product for cataract. 
     Regarding the antioxidant composition of the present invention, the pharmaceutical product for cataract may be a therapeutic agent for cataract or a prophylactic agent for cataract. 
     The antioxidant composition of the present invention may be used for an eye drop for cataract. 
     The cataract may be senile cataract. 
     The present invention provides a method of improving a skin condition comprising a step of administering an antioxidant composition comprised of one or more compounds selected from the group consisting of D- and/or L-aspartic acid, derivatives and/or salts thereof. 
     The skin condition that is suppressed and/or improved by the method of the present invention comprises, but not limited thereto, fine wrinkles, a rough skin, a dry skin, a skin cancer, a skin allergy, an inflammation of the skin, and a photosensitive dermatosis. 
     Regarding the method of the present invention, the antioxidant composition of the present invention may be used for an external preparation for the skin. 
     Regarding the method of the present invention, the antioxidant composition of the present invention may be used for a food composition. 
     The present invention also provides a method of treating and/or preventing cataract comprising a step of administering a composition comprised of one or more compounds selected from the group consisting of D- and/or L-aspartic acid, derivatives and/or salts thereof. 
     Regarding the method of treating and/or preventing cataract of the present invention, the pharmaceutical product for cataract may be an eye drop. 
     Regarding the method of treating and/or preventing cataract of the present invention, the cataract may be senile cataract. 
     As used in the present description, the term “salt” of aspartic acid indicates any salt comprising a metal salt and an amine salt and the like, provided that the antioxidant effect of aspartic acid is not impaired. The metal salt may comprise an alkaline metal salt, an alkaline earth metal salt and the like. The amine salt may comprise a triethylamine salt, a benzylamine salt and the like. 
     As used in the present description, the term “derivatives” of aspartic acid indicates an aspartic acid molecule that is covalently bound to any atomic group via its amino group, carboxyl group, or side chain, provided that the antioxidant effect of aspartic acid is not impaired. The atomic group comprises, but is not limited to, a protective group, such as N-phenylacetyl group, and 4,4′-dimethoxytrityl (DMT) group, a biopolymer, such as a protein, a peptide, a saccharide, a lipid, and a nucleic acid; a synthetic polymer, such as a polystyrene, a polyethylene, a polyvinyl, and a polyester; and a functional group such as an ester group. The ester group may comprise, for example, an aliphatic ester, such as methyl ester, and ethyl ester, and an aromatic ester. 
     An amino acid has optical isomers which are the L-form and D-form. A natural protein has L-amino acids bound by peptide bonds and only L-amino acids are employed excluding some exceptions such as a bacterial cell wall. Therefore, it has been considered that in a mammal including a human only L-amino acids are present and only L-amino acids are utilized (Kinouchi, T. et al., TANPAKUSHITSU KAKUSAN KOSO (PROTEIN, NUCLEIC ACID AND ENZYME), 50:453-460 (2005), Lehninger Principles of Biochemistry [Vol. 1] 2nd ed., pp 132-147 (1993), Japanese-language translation, Hirokawa Shoten Ltd., Harper&#39;s Biochemistry, Original version, 22nd ed., pp 21-30 (1991), Japanese-language translation Maruzen Co., Ltd.). Accordingly, only L-amino acids have been mostly employed as amino acids academically and industrially for a long time. 
     Exceptional cases where a D-amino acid is employed are, for example, a case of using as a raw material for an antibiotics produced by a microorganism, and, a case of a food additive employing a D-amino acid in a DL-amino acid mixture for the purpose of reducing cost of fractionating only an L-amino acid from a mixture of the L- and D-amino acids, which are obtained in equimolar amounts by synthesizing the amino acids. Nevertheless, there has been no case of using only D-amino acid industrially as a bioactive substance. 
     D-aspartic acid is found to be localized in the testis or the pineal body, and it is known to be involved in the control of hormone secretion (Japanese Patent Unexamined Publication No. 2005-3558). However, physiological activity of D-aspartic acid in the skin has not been clearly elucidated. 
     As shown in the following Examples, it has not been known that L- and D-aspartic acids can suppress oxidative damages so far. Thus, the antioxidant composition comprising L- and/or D-aspartic acid according to the present invention is a novel invention. 
     Recently, it was reported that ddY mice were allowed to ingest freely a 10 mM aqueous solution of a D-amino acid for two weeks and then examined for the D-amino acid concentration in each organ, which was 3 to 1000 pmol per gland in the pineal body and 2 to 500 nmol per wet gram in the brain tissue (Morikawa, A. et al., Amino Acids, 32:13-20 (2007)). Based on this, the lower limit for daily intake amount of L- and D-aspartic acids that are contained in the composition of the present invention is calculated as described below. 
     The aspartic acid of the present invention has an effect on suppressing oxidative damages in cultured human fibroblast cells at the concentrations of 0.1 μM (micro-molar) to 10 μM (micro-molar), as shown in the following examples. Thus, the amount of the aspartic acid that is contained in the agent for improving skin conditions, an external preparation for skin, and a food composition of the present invention may be any content, provided that the aspartic acid in the above concentration range is distributed to fibroblast cells in a skin tissue of a living organism. When the composition of the present invention is an external preparation, the content of the aspartic acid may be 0.000015% by weight to 50% by weight, or up to the maximum weight concentration that can be formulated, in the total composition of the present invention. Specifically, when the composition is an external preparation, the content of the aspartic acid is preferably 0.00003% by weight to 30% by weight, and the most preferably 0.0003% by weight to 3% by weight. When the composition of the present invention is an internal agent, the content of the aspartic acid may be 0.00001% by weight to 100% by weight. When the composition of the present invention is an internal agent, the content of the aspartic acid is preferably 0.00002% by weight to 80% by weight and the most preferably 0.0002% by weight to 60% by weight. Further, the lower limit of a daily intake amount of D-aspartic acid that is contained in the composition of the present invention may be 0.01 ng, preferably 0.1 ng, and more preferably 1 ng per 1 kg of body weight. The lower limit of a daily intake amount of L-aspartic acid that is contained in the composition of the present invention is an amount that is less than the general dose of commercially available medicines (20 mg per 1 kg of body weight), e.g., 0.01 mg, preferably 0.1 mg, and more preferably 1 mg per 1 kg of body weight. 
     The composition of the present invention may further comprise, in addition to the single body of aspartic acid, a salt of aspartic acid and/or a derivative capable of releasing aspartic acid by a drug metabolizing enzyme and the like in vivo, one or more pharmaceutically acceptable additives, provided that the antioxidative effect of aspartic acid on oxidative damages is not impaired. Such an additive comprises, but is not limited to, a diluent and an extender, a binder and an adhesive, a lubricant, a glidant, a plasticizer, a disintegrant, a carrier solvent, a buffering agent, a colorant, a flavoring agent, a sweetener, a preservative and a stabilizer, an adsorbent, as well as other pharmaceutical additives known to those skilled in the art. 
     The composition of the present invention may be prepared by only using, as an active ingredient, aspartic acid, salts of aspartic acid and/or derivatives capable of releasing aspartic acid by a drug metabolizing enzyme and the like in vivo. However, within the range that the effect on oxidative damages of the present invention is not impaired, it may be appropriately formulated with other components that are used for an external preparation for the skin like cosmetics including quasi drugs and pharmaceutical products, if necessary. Examples of other ingredients (i.e., optionally formulated ingredients) comprise an oil, a surface active agent, a powder, a colorant, water, alcohols, a thickening agent, a chelating agent, silicones, an antioxidant, an UV absorbing agent, a moisturizing agent, a flavoring agent, various pharmaceutically active ingredients, a preservative, a pH adjusting agent, and a neutralizing agent. 
     A dosage form of an antioxidant composition of the present invention that is used for suppressing and/or improving skin conditions (herein below, referred to as an “agent for improving skin conditions”) may be any one that is commonly used for a quasi drug composition and a pharmaceutical composition comprising an external preparation for skin like an ointment, a cream, an emulsion, a lotion, a pack, gel, and a patch; and an oral preparation like a powder, granules, a soft capsule, and a tablet; a nasal preparation like a nasal spray; and an injection solution. 
     A dosage form of the external preparation for the skin according to the present invention is not specifically limited, provided that it is conventionally used for an external preparation for the skin, and it comprises an ointment, a cream, an emulsion, a lotion, a pack, a gel, and a patch. 
     The food composition of the present invention may further comprise, in addition to aspartic acid, salts of aspartic acid and/or derivatives capable of releasing aspartic acid by a drug metabolizing enzyme and the like in vivo, a flavoring agent, a colorant, a preservative, and other components that can be used for a food product, provided that the effect of aspartic acid on oxidative damages is not impaired. 
     The food composition of the present invention may be any one employed conventionally as a food composition comprising, but not limited to, a candy, a cookie, bean paste, a French dressing, a mayonnaise, a French bread, a soy sauce, yogurt, dried seasoning powder for rice, seasoning sauce/sauce for natto (Japanese fermented soybean), natto, unrefined black vinegar. 
     ROS are known to cause not only skin disorders but also cataract. They are believed not only to disturb lipid constitution by producing lipid peroxides to polyunsaturated fatty acids inside crystalline lens via reduction of hydrogen peroxide and free radical chain reactions, but also to impair the membrane function by denaturing proteins eventually causing opacity of the lens. (“SUISHOTAI SONO SEIKAGAKUTEKI KIKO (BIOCHEMICAL MECHANISM OF CRYSTALLINE LENS”, pp 318-323, by Maesato Takami and Iwata Shuzo, edited by Iwata Shuzo, published by Medical-Aoi Publications, Inc., Tokyo (1986)). According to the findings above and the Examples described below, D- and/or L-aspartic acid having an antioxidant activity is effective for prophylaxis or treatment of cataract. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a graph showing the effect of carnosine on oxidative damages induced by hydrogen peroxide in normal human skin fibroblast cells. 
         FIG. 2  is a graph showing the effect of L-aspartic acid on oxidative damages induced by hydrogen peroxide in normal human skin fibroblast cells. 
         FIG. 3  is a graph showing the effect of D-aspartic acid on oxidative damages induced by hydrogen peroxide in normal human skin fibroblast cells. 
         FIG. 4  is a graph showing the effect of carnosine on oxidative damages by AAPH in normal human skin fibroblast cells. 
         FIG. 5  is a graph showing the effect of L- and D-aspartic acids on oxidative damages induced by AAPH in normal human skin fibroblast cells. 
     
    
    
     DESCRIPTION OF EMBODIMENTS 
     Examples of the present invention described below are intended only to exemplify the present invention rather than to limit the technical scope thereof. The technical scope of the present invention is limited only by the description in claims. 
     Example 1 
     Experiment for Evaluation of Antioxidant Effect 
     1. Object 
     ROS include reactive oxygen species in the narrow sense comprising superoxide anion, hydroxyl radical, hydroperoxide and singlet oxygen, and reactive oxygen species in the broad sense comprising an alkoxy radical, hydroperoxyl radical, a peroxyl radical, hydroperoxide, and a transition metal-oxygen complex and the like. Among the ROS, hydroxyl radical has the most potent oxidizing activity, but has very short lifetime. As such, it oxidizes non-selectively body components like nucleic acids, proteins, and lipids that are present in the vicinity of their generation site. However, peroxyl radical has a weak oxidizing activity but is relatively stable. As such, it can diffuse and cause a cell membrane damage via free radical chain reactions of polyunsaturated fatty acids. Meanwhile, hydroxyl radical generates a peroxyl radical but no hydroxyl radical is generated from a peroxyl radical. Since the working mechanism is different between a hydroxyl radical and a peroxyl radical, an effective antioxidant may be also different for each of them. For such reasons, in the present examples, an antioxidant effect was evaluated for both hydrogen peroxide and 2,2′-azobis (2-amidinopropane) dihydrochloride salt (herein below, referred to as “AAPH”), that are the representative examples of a compound which can generate hydroxyl radical and a peroxyl radical, respectively. As a positive control, carnosine having a known antioxidant activity was used. 
     2. Materials and Methods 
     2-1. Cells 
     For the evaluation of an antioxidant effect on hydrogen peroxide, human neonatal skin fibroblast cells (trade name: Cryo NHDF-Neo, manufactured by Sankyo Junyaku Co., Ltd.) were inoculated to a 24-well plate to have 1×10 5  cells per well. The cells were then cultured for four hours in a medium for cell culture (trade name: D-MEM (1 g/L glucose), manufactured by Wako Pure Chemical Industries) supplemented with 10% bovine fetal serum (herein below, referred to as a “standard medium”) in a 5% CO 2  and saturated water vapor atmosphere at 37° C. (degrees Celsius). For the evaluation of an antioxidant effect on AAPH, antibiotics (penicillin, streptomycin, and fungizone) were supplemented to the standard medium and the cells were cultured for one day. 
     2-2. Medium for Evaluation of Antioxidant Effect 
     Subsequently, the culture medium was switched to a medium for cell culture (trade name: D-MEM (1 g/L glucose), manufactured by Wako Pure Chemicals Industries) supplemented with 0.5% bovine fetal serum (herein below, referred to as “low-serum medium”) to which 0.01% or 0.05% carnosine, or 0.1 μM (micro-molar) or 10 μM (micro-molar) D- or L-aspartic acid had been added, and the cells were cultured for two days under atmosphere of 5% CO 2  and saturated moisture at 37° C. (degrees Celsius). For the experiment for the evaluation of oxidative damages on AAPH, the culture medium was switched to the medium for cell culture to which 5 ppm or 100 ppm carnosine, or 10 μM (micro-molar) D- or L-aspartic acid had been further added, and the cells were cultured for two days. The low-serum medium described above to which neither carnosine nor aspartic acid had been added was employed as a negative control. 
     2-3. Addition of Oxidant 
     After culturing for two days, 1 mM or 4 mM hydrogen peroxide, or 50 mM or 100 mM AAPH was added to the medium for evaluating an antioxidant effect, and the antioxidant effect was evaluated for carnosine or aspartic acid. The low-serum medium described above to which neither hydrogen peroxide nor AAPH had been added was employed as a control for evaluation of toxicity of an antioxidant when no oxidant was added. 
     2-4. Quantification of Oxidative Damages 
     Two hours after the addition of hydrogen peroxide or AAPH, AlarmarBlue (trade name: Biosource, manufactured by Biosource International Inc.) was added to have final concentration of 10%. Two to three hours later, according to the methods by Ahmed S. A. et al. (J. Immunol. Method., 170, 211-224 (1994)) and the instructions provided by the manufacturer, the fluorescent intensity of the supernatant was measured with an excitation wavelength of 544 nm, and an emission wavelength of 590 nm. 
     3. Results 
     3-1. Antioxidant Effect of Carnosine on Hydrogen Peroxide 
       FIG. 1  shows the results of the experiment obtained by examining the antioxidant effect of carnosine on hydrogen peroxide in Cryo NHDF-Neo cells. The error bars for each experimental condition indicate the standard deviations of the experimentally measured values obtained by repeating the experiment three times under the identical condition. The asterisk (*) indicates that t is less than 5% by Bonferroni/Dunn test. 
     The ratio of viable cells for the control group for evaluating toxicity of the antioxidant without addition of an oxidant was 102% when carnosine had not been added. When the concentration of carnosine was 0.01%, it was 100%. When the concentration of carnosine was 0.05%, it was 97%. The ratio of viable cells in the case of addition of 1 mM hydrogen peroxide was 45% when carnosine had not been added. When the concentration of carnosine was 0.01%, it was 51%. When the concentration of carnosine was 0.05%, it was 53%. The ratio of viable cells in the case of 4 mM hydrogen peroxide was 14% when carnosine had not been added. When the concentration of carnosine was 0.01%, it was 21%. When the concentration of carnosine was 0.05%, it was 45%. Thus, when the concentration of hydrogen peroxide was 4 mM, a significant difference in the ratio of viable cells was observed for the case in which 0.05% carnosine had been added compared to the ratio without addition of carnosine. Based on the results above, the antioxidant effect of carnosine on hydrogen peroxide was confirmed in the experiment system of the present example. 
     3-2. Antioxidant Effect of L-Aspartic Acid on Hydrogen Peroxide 
       FIG. 2  shows the results of the experiment obtained by examining the antioxidant effect of L-aspartic acid on hydrogen peroxide in Cryo NHDF-Neo cells. The error bars for each experimental condition indicate the standard deviations of experimentally measured values obtained by repeating the experiment three times under the identical condition. 
     The ratio of viable cells for the control group for evaluating toxicity of an antioxidant without addition of an oxidant was 93% when L-aspartic acid had not been added. When the concentration of L-aspartic acid was 0.1 μl (micro-molar), it was 88%. When the concentration of L-aspartic acid was 10 μM (micro-molar), it was 97%. The ratio of viable cells in the case of 1 mM hydrogen peroxide was 61% when L-aspartic acid had not been added. When the concentration of L-aspartic acid was 0.1 μM (micro-molar), it was 62%. When the concentration of L-aspartic acid was 10 μM (micro-molar), it was 62%. The ratio of viable cells in the case of 4 mM hydrogen peroxide was 36% when L-aspartic acid had not been added. When the concentration of L-aspartic acid was 0.1 μM (micro-molar), it was 33%. When the concentration of L-aspartic acid was 10 μM (micro-molar), it was 32%. Based on the results above, a statistically significant antioxidant effect of L-aspartic acid on hydrogen peroxide was not observed. 
     3-3. Antioxidant Effect of D-Aspartic Acid on Hydrogen Peroxide 
       FIG. 3  shows the results of the experiment obtained by examining the antioxidant effect of D-aspartic acid on oxidative damages induced by hydrogen peroxide in Cryo NHDF-Neo cells. The error bars for each experimental condition indicate the standard deviations of experimentally measured values obtained by repeating the experiment three times under the identical condition. The asterisk (*) indicates that p is less than 5% by Bonferroni/Dunn test. The double asterisk (**) indicates that p is less than 1% by Bonferroni/Dunn test. 
     The ratio of viable cells for the control group for evaluating toxicity of an antioxidant without addition of an oxidant was 97% when D-aspartic acid had not been added. When the concentration of D-aspartic acid was 0.1 μM (micro-molar), it was 86%. When the concentration of D-aspartic acid was 10 μM (micro-molar), it was 97%. In the case of 1 mM hydrogen peroxide, the ratio of viable cells was 55% when D-aspartic acid had not been added. When the concentration of D-aspartic acid was 0.1 μM (micro-molar), it was 62%. When the concentration of D-aspartic acid was 10 μM (micro-molar), it was 63%. In the case of 4 mM hydrogen peroxide, the ratio of viable cells was 22% when D-aspartic acid had not been added. When the concentration of D-aspartic acid was 0.1 μM (micro-molar), it was 29%. When the concentration of D-aspartic acid was 10 μM (micro-molar), it was 34%. Thus, when the concentration of hydrogen peroxide was 4 mM, a significant difference in the ratio of viable cells was observed for the case in which 0.1 μM (micro-molar) or 10 μM (micro-molar) D-aspartic acid had been added compared to the ratio without addition of D-aspartic acid. Based on the results above, the concentration-dependent antioxidant effect of D-aspartic acid on hydrogen peroxide was observed. 
     3-4. Antioxidant Effect of Carnosine in Experiment for Evaluating Oxidative Damages Induced by AAPH 
       FIG. 4  shows the experiment obtained by examining the antioxidant effect of carnosine in the experiment for evaluating oxidative damages induced by AAPH in Cryo NHDF-Neo cells. The error bars for each experimental condition indicate the standard deviations of experimentally measured values obtained by repeating the experiment three times under the identical condition. The double asterisk (**) indicates that p is less than 1% by Bonferroni/Dunn test. 
     The ratio of viable cells for the control group for evaluating toxicity of an antioxidant without addition of an oxidant was 100% when carnosine had not been added. When the concentration of carnosine was 5 ppm, it was 93%. When the concentration of carnosine was 100 ppm, it was 103%. The ratio of viable cells in the case of 100 mM AAPH was 31% when carnosine had not been added. When the concentration of carnosine was 5 ppm, it was 60%. When the concentration of carnosine was 100 ppm, it was 85%. When AAPH had concentration of 100 mM, a significant difference was observed for the ratio of viable cells for the case in which carnosine had been added at the concentration of 100 ppm compared to the ratio without addition of carnosine. Based on the results above, the antioxidant effect of carnosine on AAPH was confirmed by the experiment system of the present example. 
     3-5. Antioxidant Effect of L- and D-Aspartic Acids in Experiment for Evaluating Oxidative Damages Induced by AAPH 
       FIG. 5  shows the experiment obtained by examining the antioxidant effect of L- and D-aspartic acids in the experiment for evaluating oxidative damages induced by AAPH in Cryo NHDF-Neo cells. The error bars for each experimental condition indicate the standard deviations of the experimentally measured values obtained by repeating the test three times under the identical condition. The asterisk (*) indicates that p is less than 5% by Bonferroni/Dunn test. The triple asterisk (***) indicates that p is less than 0.1% by Bonferroni/Dunn test. 
     The ratio of viable cells for the control group for evaluating toxicity of an antioxidant without addition of the oxidant was 95% when L- and D-aspartic acids had not been added. When the concentration of D-aspartic acid was 10 μM (micro-molar), it was 102%. When the concentration of L-aspartic acid was 10 μM (micro-molar), it was 80%. In the case of 100 mM AAPH, the ratio of viable cells was 51% when L- and D-aspartic acids had not been added. When the concentration of D-aspartic acid was 10 μM (micro-molar), it was 96%. When the concentration of L-aspartic acid was 10 μM (micro-molar), it was 69%. When AAPH had concentration of 100 mM, a significant difference was observed for the ratio of viable cells for the case in which L- and D-aspartic acids had been added at concentration of 10 μM (micro-molar) compared to the ratio without addition of L- and D-aspartic acids. Based on the results above, it was indicated that D-aspartic acid had more potent antioxidant effect on AAPH compared to L-aspartic acid. 
     4. Conclusions 
     Based on the experimental results of the Examples above, D-aspartic acid was found to have an antioxidant effect on both hydrogen peroxide and AAPH. However, L-aspartic acid was found to have an antioxidant effect on AAPH only. Thus, it was indicated that D-aspartic acid is effective against both hydroxyl radical and a peroxyl radical, but L-aspartic acid is effective only against a peroxyl radical. 
     Example 2 
     Formulation examples of a composition comprising aspartic acid according to the present invention, i.e., an emulsion preparation, a patch, a tablet, a soft capsule, a granule, a beverage, a candy, a cookie, bean paste, a French dressing, a mayonnaise, a French bread, a soy sauce, yogurt, dried seasoning powder for rice, seasoning/sauce for natto, natto, unrefined black vinegar, cream, body cream, gel, a peel-off mask, a wet pack, an emulsion, a skin lotion, and an aerosol preparation, are given below. The aspartic acid in the formulation examples is either D-form and/or L-form. 
     These formulation examples are all illustrative and not intended to limit the technical scope of the present invention. 
     Formulation Example 1 
     Emulsion Preparation 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Content 
               
               
                   
                 (Composition) 
                 (% by weight) 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Aspartic acid 
                 0.42 
               
               
                   
                 Behenyl alcohol 
                 0.2 
               
               
                   
                 Cetanol 
                 0.5 
               
               
                   
                 Glycerin monofatty acid ester 
                 1.8 
               
               
                   
                 Hydrogenated castor oil POE 
                 1.0 
               
               
                   
                 (60) 
               
               
                   
                 White petrolatum 
                 2.0 
               
               
                   
                 Liquid paraffin 
                 10.0 
               
               
                   
                 Isopropyl myristate 
                 3.0 
               
               
                   
                 Methyl polysiloxane (6cs) 
                 1.5 
               
               
                   
                 Concentrated glycerin 
                 13.0 
               
               
                   
                 Dipropylene glycol 
                 2.0 
               
               
                   
                 Carboxyvinyl polymer 
                 0.25 
               
               
                   
                 Sodium hyaluronate 
                 0.005 
               
               
                   
                 Potassium hydroxide 
                 Proper quantity 
               
               
                   
                 Lactic acid 
                 Proper quantity 
               
               
                   
                 Edetate sodium 
                 Proper quantity 
               
               
                   
                 Ethylparaben 
                 Proper quantity 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.000 
               
               
                   
                   
               
             
          
         
       
     
     Formulation Example 2 
     Patch 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (% by weight) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Aspartic acid 
                 0.3 
               
               
                   
                 Polyacrylic acid 
                 3.0 
               
               
                   
                 Sodium polyacrylate 
                 2.5 
               
               
                   
                 Gelatin 
                 0.5 
               
               
                   
                 Sodium carboxymethyl cellulose 
                 4.0 
               
               
                   
                 Polyvinyl alcohol 
                 0.3 
               
               
                   
                 Concentrated glycerin 
                 14.0 
               
               
                   
                 1,3-Butylene glycol 
                 12.0 
               
               
                   
                 Aluminum hydroxide 
                 0.1 
               
               
                   
                 Edetate sodium 
                 0.03 
               
               
                   
                 Methylparaben 
                 0.1 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 3 
     Tablet 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (mg/tablet) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Aspartic acid 
                 360.5 
               
               
                   
                 Lactose 
                 102.4 
               
               
                   
                 Calcium carboxymethyl 
                 29.9 
               
               
                   
                 cellulose 
                   
               
               
                   
                 Hydroxypropyl cellulose 
                 6.8 
               
               
                   
                 Magnesium stearate 
                 5.2 
               
               
                   
                 Crystalline cellulose 
                 10.2 
               
               
                   
                   
                 515.0 
               
               
                   
               
             
          
         
       
     
     Formulation Example 4 
     Tablet 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (mg/tablet) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Sucrose ester 
                 70 
               
               
                   
                 Crystalline cellulose 
                 74 
               
               
                   
                 Methyl cellulose 
                 36 
               
               
                   
                 Glycerin 
                 25 
               
               
                   
                 Aspartic acid 
                 475 
               
               
                   
                 N-Acetylglucosamine 
                 200 
               
               
                   
                 Hyaluronic acid 
                 150 
               
               
                   
                 Vitamin E 
                 30 
               
               
                   
                 Vitamin B6 
                 20 
               
               
                   
                 Vitamin B2 
                 10 
               
               
                   
                 α(alpha)-Lipoic acid 
                 20 
               
               
                   
                 Coenzyme Q10 
                 40 
               
               
                   
                 Ceramide ( Konjac  extract) 
                 50 
               
               
                   
                 L-Proline 
                 300 
               
               
                   
                   
                 1500 
               
               
                   
               
             
          
         
       
     
     Formulation Example 5 
     Soft Capsule 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (mg/capsule) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Edible soybean oil 
                 530 
               
               
                   
                   Eucommia ulmoides  extract 
                 50 
               
               
                   
                 Ginseng extract 
                 50 
               
               
                   
                 Aspartic acid 
                 100 
               
               
                   
                 Royal jelly 
                 50 
               
               
                   
                 Maca 
                 30 
               
               
                   
                 GABA 
                 30 
               
               
                   
                 Beeswax 
                 60 
               
               
                   
                 Gelatin 
                 375 
               
               
                   
                 Glycerin 
                 120 
               
               
                   
                 Glycerin fatty acid ester 
                 105 
               
               
                   
                   
                 1500 
               
               
                   
               
             
          
         
       
     
     Formulation Example 6 
     Soft Capsule 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (mg/capsule) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Brown rice germ oil 
                 659 
               
               
                   
                 Aspartic acid 
                 500 
               
               
                   
                 Resveratrol 
                 1 
               
               
                   
                 Lotus germ extract 
                 100 
               
               
                   
                 Elastin 
                 180 
               
               
                   
                 DNA 
                 30 
               
               
                   
                 Folic acid 
                 30 
               
               
                   
                   
                 1500 
               
               
                   
               
             
          
         
       
     
     Formulation Example 7 
     Granule 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (mg/pack) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Aspartic acid 
                 400 
               
               
                   
                 Vitamin C 
                 100 
               
               
                   
                 Soybean isoflavone 
                 250 
               
               
                   
                 Reduced lactose 
                 300 
               
               
                   
                 Soybean oligosaccharide 
                 36 
               
               
                   
                 Erythritol 
                 36 
               
               
                   
                 Dextrin 
                 30 
               
               
                   
                 Flavoring agent 
                 24 
               
               
                   
                 Citric acid 
                 24 
               
               
                   
                   
                 1200 
               
               
                   
               
             
          
         
       
     
     Formulation Example 8 
     Beverage 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (g/60 mL) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   Eucommia ulmoides  extract 
                 1.6 
               
               
                   
                 Ginseng extract 
                 1.6 
               
               
                   
                 Aspartic acid 
                 0.25 
               
               
                   
                 Reduced maltose syrup 
                 28 
               
               
                   
                 Erythritol 
                 8 
               
               
                   
                 Citric acid 
                 2 
               
               
                   
                 Flavoring agent 
                 1.3 
               
               
                   
                 N-Acetylglucosamine 
                 1 
               
               
                   
                 Sodium hyaluronate 
                 0.5 
               
               
                   
                 Vitamin E 
                 0.3 
               
               
                   
                 Vitamin B6 
                 0.2 
               
               
                   
                 Vitamin B2 
                 0.1 
               
               
                   
                 α (alpha)-Lipoic acid 
                 0.2 
               
               
                   
                 Coenzyme Q10 
                 1.2 
               
               
                   
                 Ceramide ( Konjac  extract) 
                 0.4 
               
               
                   
                 L-Proline 
                 2 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 60 
               
               
                   
               
             
          
         
       
     
     Formulation Example 9 
     Candy 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (% by weight) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Sugar 
                 50 
               
               
                   
                 Syrup 
                 48 
               
               
                   
                 Aspartic acid 
                 1 
               
               
                   
                 Flavoring agent 
                 1 
               
               
                   
                   
                 100 
               
               
                   
               
             
          
         
       
     
     Formulation Example 10 
     Cookie 
                                                       (Composition)   Content (% by weight)                                    Weak flour   45.0           Butter   17.5           Granulated sugar   20.0           Aspartic acid   4.0           Egg   12.5           Flavoring agent   1.0               100.0                    
Method for Producing Formulation Example 10 (Cookie)
 
     Granulated sugar is added in portions to butter while stirring, to which an egg, aspartic acid and a flavoring agent are added and stirred. After mixing thoroughly, uniformly sieved weak flour is added and stirred at a low speed, and allowed to stand as a bulk in a refrigerator. Thereafter, it is molded and baked for 15 minutes at 170° C. (degrees Celsius) to obtain a cookie. 
     Formulation Example 11 
     Bean Paste 
                                                       (Composition)   Content (g)                                    Soybean   1000           Malted rice   1000           Salt   420           Aspartic acid   16.8           Water   Remainder               4000                    
Method for Producing Formulation Example 11 (Bean Paste)
 
     Malted rice is mixed thoroughly with a salt. Washed soybeans are soaked overnight in three times its volumes of water, which are then drained off, and new water is added while boiling, and poured into a colander to collect the broth (tanemizu fluid), to which aspartic acid is dissolved at 10% w/v. The boiled beans are minced immediately, combined with malted rice mixed with salt, to which the tanemizu fluid containing aspartic acid dissolved therein is added and kneaded evenly to obtain a clay-like hardness. Dumplings are made and stuffed in a container compactly without forming any voids, and the surface of the content is smoothened and sealed with a plastic film. After three months, the content is transferred to a new container and the surface is smoothened and sealed with a plastic film. Instead of adding aspartic acid to the tanemizu fluid, a malted rice producing a large amount of aspartic acid may be employed. Such malted rice can be selected by quantifying aspartic acid by the method described in Japanese Patent Unexamined Publication No. 2008-185558. Alternatively, a commercially available bean paste can be supplemented with aspartic acid or a salt thereof. 
     Formulation Example 12 
     French Dressing 
                                                       (Composition)   Content (g)                                    Salad oil   27.4           Vinegar   30.4           Sodium chloride   0.9           Aspartic acid   0.30           Pepper   1.0               60.0                    
Method for Producing Formulation Example 12 (French Dressing)
 
     Vinegar is combined with sodium chloride and aspartic acid, and then stirred thoroughly to be dissolved. Salad oil is added to the mixture and the mixture is stirred thoroughly and then pepper is added. 
     Formulation Example 13 
     Mayonnaise 
                                                       (Composition)   Content (g)                                    Salad oil   134.0           Vinegar   5.5           Sodium chloride   0.9           Aspartic acid   0.5           Egg yolk   18           Sugar   0.2           Pepper   0.9               160.0                    
Method for Producing Formulation Example 13 (Mayonnaise)
 
     An egg yolk (room temperature) is combined with vinegar, sodium chloride, aspartic acid and pepper, and stirred thoroughly using a whipping apparatus. Stirring is continued while adding salad oil in portions to form an emulsion. Finally, sugar is added and the mixture is stirred. 
     Formulation Example 14 
     French Bread 
                                                       (Composition)   Content (g)                                    Hard flour   140           Weak flour   60           Sodium chloride   3           Sugar   6           Aspartic acid   2           Dry yeast   4           Lukewarm water   128               343                    
Method for Producing Formulation Example 14 (French Bread)
 
     Lukewarm water is combined with 1 g of sugar and dry yeast, which is then allowed to undergo a pre-fermentation. Hard flour, weak flour, sodium chloride, 5 g of sugar and aspartic acid are placed in a bowl, into which the pre-fermented yeast is placed. After kneading thoroughly into a ball-like dough, a primary fermentation is conducted at 30° C. (degrees Celsius). The dough is kneaded again and allowed to stand, and then shaped into suitable forms, which are subjected to a final fermentation using an electronic fermentation machine. After forming coupes, baking is conducted for 30 minutes in an oven at 220° C. (degrees Celsius). 
     Formulation Example 15 
     Soy Sauce 
                                                       (Composition)   Content (g)                                    Commercially available soy   995.8           sauce               Aspartic acid   4.2               1000                    
Method for Producing Formulation Example 15 (Soy Sauce)
 
     Commercially available soy sauce is supplemented with aspartic acid, and stirred thoroughly. Instead of adding aspartic acid or a salt thereof, malted rice producing a large amount of aspartic acid may be employed for fermenting soy sauce. Such malted rice can be selected by quantifying aspartic acid by the method described in Japanese Patent Unexamined Publication No. 2008-185558. Further, aspartic acid or a salt thereof may be added to commercially available soy sauce. 
     Formulation Example 16 
     Yogurt 
                                                       (Composition)   Content (g)                                    Milk   880             L. bulgaricus     50             S. thermophilus     50           Aspartic acid   20               1000                    
Method for Producing Formulation Example 16 (Yogurt)
 
     Fermentation is conducted at 40 to 45° C. (degrees Celsius). Other commercially available fermentation seed organisms may be employed and commercially available yogurt may be supplemented with aspartic acid. Instead of adding aspartic acid or a salt thereof, a seed organism producing a large amount of aspartic acid may be employed. Such an organism can be selected by quantifying aspartic acid by the method described in Japanese Patent Unexamined Publication No. 2008-185558. Further, aspartic acid or a salt thereof may be added to commercially available yogurt. 
     Formulation Example 17 
     Dried Seasoning Powder for Rice 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (g) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Aspartic acid 
                 50 
               
               
                   
                 Laver 
                 15 
               
               
                   
                 Sodium L-glutamate 
                 10 
               
               
                   
                 Sodium chloride 
                 2 
               
               
                   
                 Roasted sesame 
                 10 
               
               
                   
                 Dried mackerel shavings 
                 10 
               
               
                   
                 Sugar 
                 1 
               
               
                   
                 Soy sauce 
                 2 
               
               
                   
                   
                 100 
               
               
                   
               
             
          
         
       
     
     Formulation Example 18 
     Seasoning, Sauce for Natto 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (g) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Commercially available sauce 
                 9.96 
               
               
                   
                 for natto 
                   
               
               
                   
                 Aspartic acid 
                 0.04 
               
               
                   
                   
                 10 
               
               
                   
               
             
          
         
       
     
     Formulation Example 19 
     Natto 
                                                       (Composition)   Content (g)                                    Commercially available natto   19.9           Aspartic acid   0.1               20                    
Method for Producing Formulation Example 19 (Natto)
 
     Instead of adding aspartic acid or a salt thereof, an organism producing a large amount of aspartic acid may be employed for producing natto. Such an organism can be selected by quantifying aspartic acid by the method described in Japanese Patent Unexamined Publication No. 2008-185558. Further, aspartic acid or a salt thereof may be added to commercially available natto. 
     Formulation Example 20 
     Unrefined Black Vinegar 
                                                       (Composition)   Content (g)                                    Commercially available   995.8           unrefined black vinegar               Aspartic acid   4.2               1000                    
Method for Producing Formulation Example 20 (Unrefined Black Vinegar)
 
     Instead of adding aspartic acid or a salt thereof, an organism producing a large amount of aspartic acid may be employed for producing vinegar, black vinegar or unrefined vinegar. Such an organism can be selected by quantifying aspartic acid by the method described in Japanese Patent Unexamined Publication No. 2008-185558. Further, aspartic acid or a salt thereof may be added to commercially available unrefined black vinegar. 
     Formulation Example 21 
     Cream 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Liquid paraffin 
                 3 
               
               
                   
                 White petrolatum 
                 1 
               
               
                   
                 Dimethyl polysiloxane 
                 1 
               
               
                   
                 Stearyl alcohol 
                 1.8 
               
               
                   
                 Behenyl alcohol 
                 1.6 
               
               
                   
                 Glycerin 
                 8 
               
               
                   
                 Dipropylene glycol 
                 5 
               
               
                   
                 Macadamia nut oil 
                 2 
               
               
                   
                 Hydrogenated oil 
                 3 
               
               
                   
                 Squalane 
                 6 
               
               
                   
                 Stearic acid 
                 2 
               
               
                   
                 Cholesteryl hydroxystearate 
                 0.5 
               
               
                   
                 Cetyl 2-ethylhexanoate 
                 4 
               
               
                   
                 Polyoxyethylene hydrogenated 
                 0.5 
               
               
                   
                 castor oil 
                   
               
               
                   
                 Self-emulsifying glyceryl 
                 3 
               
               
                   
                 monostearate 
                   
               
               
                   
                 Potassium hydroxide 
                 0.15 
               
               
                   
                 Sodium hexametaphosphate 
                 0.05 
               
               
                   
                 Trimethyl glycine 
                 2 
               
               
                   
                 Potassium ascorbyl tocopheryl 
                 1 
               
               
                   
                 phosphate 
                   
               
               
                   
                 Tocopheryl acetate 
                 0.1 
               
               
                   
                 Aspartic acid 
                 0.42 
               
               
                   
                 Paraben 
                 Proper quantity 
               
               
                   
                 Edetate trisodium 
                 0.05 
               
               
                   
                 4-t-Butyl-4′-methoxy 
                 0.05 
               
               
                   
                 dibenzoylmethane 
                   
               
               
                   
                 Glyceryl ethylhexanoate 
                 0.05 
               
               
                   
                 dimethoxycinnamate 
                   
               
               
                   
                 Colorant 
                 Proper quantity 
               
               
                   
                 Carboxyvinyl polymer 
                 0.05 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 22 
     Body Cream 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Dimethyl polysiloxane 
                 3 
               
               
                   
                 Decamethyl cyclopentasiloxane 
                 13 
               
               
                   
                 Dodecamethyl 
                 12 
               
               
                   
                 cyclohexasiloxane 
                   
               
               
                   
                 Polyoxyethylene 
                 1 
               
               
                   
                 methylpolysiloxane copolymer 
                   
               
               
                   
                 Ethanol 
                 2 
               
               
                   
                 Isopropanol 
                 1 
               
               
                   
                 Glycerin 
                 3 
               
               
                   
                 Dipropylene glycol 
                 5 
               
               
                   
                 Polyethylene glycol 6000 
                 5 
               
               
                   
                 Sodium hexametaphosphate 
                 0.05 
               
               
                   
                 Tocopherol acetate 
                 0.1 
               
               
                   
                 Aspartic acid 
                 0.3 
               
               
                   
                   Foeniculum vulgare  (Fennel) 
                 0.1 
               
               
                   
                 extract 
                   
               
               
                   
                   Hamamelis virginiana  (Witch 
                 0.1 
               
               
                   
                 Hazel) extract 
                   
               
               
                   
                 Ginseng extract 
                 0.1 
               
               
                   
                 L-Menthol 
                 Proper quantity 
               
               
                   
                 Paraoxybenzoic acid ester 
                 Proper quantity 
               
               
                   
                 (Paraben) 
                   
               
               
                   
                 Edetate trisodium 
                 0.05 
               
               
                   
                 Dimorpholinopyridazinone 
                 0.01 
               
               
                   
                 Isopentyl trimethoxycinnamate 
                 0.1 
               
               
                   
                 trisiloxane 
                   
               
               
                   
                 Iron oxide yellow 
                 Proper quantity 
               
               
                   
                 Cobalt titanium oxide 
                 Proper quantity 
               
               
                   
                 Dimethyl distearyl ammonium 
                 1.5 
               
               
                   
                 hectorite 
                   
               
               
                   
                 Polyvinyl alcohol 
                 0.1 
               
               
                   
                 Hydroxyethylcellulose 
                 0.1 
               
               
                   
                 Trimethylsiloxysilicate 
                 2 
               
               
                   
                 Flavoring agent 
                 Proper quantity 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 23 
     Gel 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Dimethyl polysiloxane 
                 5 
               
               
                   
                 Glycerin 
                 2 
               
               
                   
                 1,3-Butylene glycol 
                 5 
               
               
                   
                 Polyethylene glycol 1500 
                 3 
               
               
                   
                 Polyethylene glycol 20000 
                 3 
               
               
                   
                 Cetyl ethylhexanoate 
                 3 
               
               
                   
                 Citric acid 
                 0.01 
               
               
                   
                 Sodium citrate 
                 0.1 
               
               
                   
                 Sodium hexametaphosphate 
                 0.1 
               
               
                   
                 Dipotassium glycyrrhizate 
                 0.1 
               
               
                   
                 Aspartic acid 
                 0.4 
               
               
                   
                 Tocopheryl acetate 
                 0.1 
               
               
                   
                   Scutellaria Baicalensis  root 
                 0.1 
               
               
                   
                 extract 
                   
               
               
                   
                   Saxifraga sarmentos  extract 
                 0.1 
               
               
                   
                 Edetate trisodium 
                 0.1 
               
               
                   
                 Xanthan gum 
                 0.3 
               
               
                   
                 Acrylates/C10-30 alkyl 
                 0.05 
               
               
                   
                 acrylate crosspolymer (Pemulen 
                   
               
               
                   
                 TR-2) 
                   
               
               
                   
                 Agar powder 
                 1.5 
               
               
                   
                 Phenoxyethanol 
                 Proper quantity 
               
               
                   
                 Dibutylhydroxytoluene 
                 Proper quantity 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 24 
     Peel-Off Mask 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Ethanol 
                 10 
               
               
                   
                 1,3-Butylene glycol 
                 6 
               
               
                   
                 Polyethylene glycol 4000 
                 2 
               
               
                   
                 Olive oil 
                 1 
               
               
                   
                 Macadamia nut oil 
                 1 
               
               
                   
                 Phytosteryl hydroxystearic 
                 0.05 
               
               
                   
                 acid 
                   
               
               
                   
                 Lactic acid 
                 0.05 
               
               
                   
                 Sodium lactate 
                 0.1 
               
               
                   
                 Disodium ascorbyl sulfate 
                 0.1 
               
               
                   
                 Potassium ascorbyl tocopheryl 
                 0.1 
               
               
                   
                 phosphate 
                   
               
               
                   
                 Aspartic acid 
                 0.3 
               
               
                   
                 Fish collagen 
                 0.1 
               
               
                   
                 Sodium chondroitin sulfate 
                 0.1 
               
               
                   
                 Sodium carboxymethyl cellulose 
                 0.2 
               
               
                   
                 Polyvinyl alcohol 
                 12 
               
               
                   
                 Paraoxybenzoic acid ester 
                 Proper quantity 
               
               
                   
                 (Paraben) 
                   
               
               
                   
                 Flavoring agent 
                 Proper quantity 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 25 
     Wet Pack 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Glycerin 
                 1 
               
               
                   
                 1,3-Butylene glycol 
                 8 
               
               
                   
                 Xylit 
                 2 
               
               
                   
                 Polyethylene glycol 1500 
                 2 
               
               
                   
                 Rosemary oil 
                 0.01 
               
               
                   
                 Sage oil 
                 0.1 
               
               
                   
                 Citric acid 
                 0.02 
               
               
                   
                 Sodium citrate 
                 0.08 
               
               
                   
                 Sodium hexametaphosphate 
                 0.01 
               
               
                   
                 Hydroxypropyl-β(beta)-cyclodextrin 
                 0.1 
               
               
                   
                 Aspartic acid 
                 0.2 
               
               
                   
                 Birch extract 
                 0.1 
               
               
                   
                 Lavender oil 
                 0.01 
               
               
                   
                 Xanthane gum 
                 0.05 
               
               
                   
                 Carboxylvinyl polymer 
                 0.15 
               
               
                   
                 Paraoxybenzoic acid ester 
                 Proper quantity 
               
               
                   
                 (Paraben) 
                   
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 26 
     Emulsion 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Liquid paraffin 
                 7 
               
               
                   
                 White petrolatum 
                 3 
               
               
                   
                 Decamethyl cyclopentasiloxane 
                 2 
               
               
                   
                 Behenyl alcohol 
                 1.5 
               
               
                   
                 Glycerin 
                 5 
               
               
                   
                 Dipropylene glycol 
                 7 
               
               
                   
                 Polyethylene glycol 1500 
                 2 
               
               
                   
                 Jojoba oil 
                 1 
               
               
                   
                 Isostearic acid 
                 0.5 
               
               
                   
                 Stearic acid 
                 0.5 
               
               
                   
                 Behenic acid 
                 0.5 
               
               
                   
                 Pentaerythritol tetra 
                 3 
               
               
                   
                 (2-ethylhexanoate) 
                   
               
               
                   
                 Cetyl 2-ethylhexanoate 
                 3 
               
               
                   
                 Glycerin monostearate 
                 1 
               
               
                   
                 Polyoxyethylene-glycerin 
                 1 
               
               
                   
                 monostearate 
                   
               
               
                   
                 Potassium hydroxide 
                 0.1 
               
               
                   
                 Sodium hexametaphosphate 
                 0.05 
               
               
                   
                 Stearyl glycyrrhetinate 
                 0.05 
               
               
                   
                 Aspartic acid 
                 0.3 
               
               
                   
                 Royal jelly extract 
                 0.1 
               
               
                   
                 Yeast extract 
                 0.1 
               
               
                   
                 Tocopherol acetate 
                 0.1 
               
               
                   
                 Acetylated sodium hyaluronate 
                 0.1 
               
               
                   
                 Edetate trisodium 
                 0.05 
               
               
                   
                 4-t-Butyl-4′-methoxydibenzoyl 
                 0.1 
               
               
                   
                 methane 
                   
               
               
                   
                 2-Ethylhexyl 
                 0.1 
               
               
                   
                 paramethoxycinnamate 
                   
               
               
                   
                 Carboxylvinyl polymer 
                 0.15 
               
               
                   
                 Paraben 
                 Proper quantity 
               
               
                   
                 Flavoring agent 
                 Proper quantity 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 27 
     Emulsion 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Dimethylpolysiloxane 
                 2 
               
               
                   
                 Behenyl alcohol 
                 1 
               
               
                   
                 Batyl alcohol 
                 0.5 
               
               
                   
                 Glycerin 
                 5 
               
               
                   
                 1,3-Butylene glycol 
                 7 
               
               
                   
                 Erythritol 
                 2 
               
               
                   
                 Hydrogenated oil 
                 3 
               
               
                   
                 Squalane 
                 6 
               
               
                   
                 Pentaerythritol tetra 
                 2 
               
               
                   
                 (2-ethylhexanoate) 
                   
               
               
                   
                 Polyoxyethylene glyceryl 
                 1 
               
               
                   
                 isostearate 
                   
               
               
                   
                 Polyoxyethylene glyceryl 
                 1 
               
               
                   
                 monostearate 
                   
               
               
                   
                 Aspartic acid 
                 0.3 
               
               
                   
                 Potassium hydroxide 
                 Proper quantity 
               
               
                   
                 Sodium hexametaphosphate 
                 0.05 
               
               
                   
                 Phenoxyethanol 
                 Proper quantity 
               
               
                   
                 Carboxylvinyl polymer 
                 0.1 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 28 
     Skin Lotion 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Ethyl alcohol 
                 5 
               
               
                   
                 Glycerin 
                 1 
               
               
                   
                 1,3-Butylene glycol 
                 5 
               
               
                   
                 Polyoxyethylene-polyoxypropylene 
                 0.2 
               
               
                   
                 decyltetradecyl ether 
                   
               
               
                   
                 Sodium hexametaphosphate 
                 0.03 
               
               
                   
                 Trimethyl glycine 
                 1 
               
               
                   
                 Sodium polyasparaginate 
                 0.1 
               
               
                   
                 Potassium ascorbyl tocopheryl 
                 0.1 
               
               
                   
                 phosphate 
                   
               
               
                   
                 Thiotaurine 
                 0.1 
               
               
                   
                 Aspartic acid 
                 0.2 
               
               
                   
                 Edetate trisodium 
                 0.1 
               
               
                   
                 Carboxylvinyl polymer 
                 0.05 
               
               
                   
                 Potassium hydroxide 
                 0.02 
               
               
                   
                 Phenoxyethanol 
                 Proper quantity 
               
               
                   
                 Flavoring agent 
                 Proper quantity 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 29 
     Skin Lotion 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (%) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Ethanol 
                 10 
               
               
                   
                 Dipropylene glycol 
                 1 
               
               
                   
                 Polyethylene glycol 1000 
                 1 
               
               
                   
                 Polyoxyethylene methyl 
                 1 
               
               
                   
                 glucoside 
                   
               
               
                   
                 Jojoba oil 
                 0.01 
               
               
                   
                 Glyceryl 
                 0.1 
               
               
                   
                 tri(2-ethylhexanoate) 
                   
               
               
                   
                 Polyoxyethylene hydrogenated 
                 0.2 
               
               
                   
                 castor oil 
                   
               
               
                   
                 Polyglyceryl diisostearate 
                 0.15 
               
               
                   
                 Sodium N-stearoyl-L-glutamate 
                 0.1 
               
               
                   
                 Citric acid 
                 0.05 
               
               
                   
                 Sodium citrate 
                 0.2 
               
               
                   
                 Potassium hydroxide 
                 0.4 
               
               
                   
                 Dipotassium glycyrrhizate 
                 0.1 
               
               
                   
                 Arginine hydrochloride 
                 0.1 
               
               
                   
                 L-Ascorbic acid 2-glucoside 
                 2 
               
               
                   
                 Aspartic acid 
                 0.2 
               
               
                   
                 Edetate trisodium 
                 0.05 
               
               
                   
                 Octyl 4-methoxycinnamate 
                 0.01 
               
               
                   
                 Dibutylhydroxytoluene 
                 Proper quantity 
               
               
                   
                 Paraben 
                 Proper quantity 
               
               
                   
                 Deep sea water 
                 3 
               
               
                   
                 Flavoring agent 
                 Proper quantity 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 30 
     Stock Solution of Aerosol Urea Preparation for External Use 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 (Composition) 
                 Content (% by weight) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Ethanol 
                 15.0 
               
               
                   
                 Polyoxyethylene hydrogenated 
                 1.5 
               
               
                   
                 castor oil 50 
                   
               
               
                   
                 Diphenhydramine 
                 1.0 
               
               
                   
                 Dibucaine 
                 2.0 
               
               
                   
                 Tocopheryl acetate 
                 0.5 
               
               
                   
                 Aspartic acid 
                 0.1 
               
               
                   
                 Isostearic acid 
                 0.1 
               
               
                   
                 1,3-Butylene glycol 
                 3.0 
               
               
                   
                 Polyethylene glycol 400 
                 3.0 
               
               
                   
                 Camphor 
                 0.05 
               
               
                   
                 Urea 
                 20.0 
               
               
                   
                 Purified water 
                 Remainder 
               
               
                   
                   
                 100.00 
               
               
                   
               
             
          
         
       
     
     Formulation Example 31 
     Aerosol Urea Spray 
                                                       (Composition)   Content (% by weight)                                    Stock solution of aerosol urea   65.0           preparation for external use               Dimethyl ether   35.0               100.00                    
Method of Filling Formulation Example 31 (Aerosol Urea Spray)
 
     Stock solution of an aerosol urea external preparation and dimethyl ether are filled in a pressure resistant aerosol aluminum can of which internal surface is coated with Teflon (registered trade mark) to prepare an aerosol preparation.