JPH05320185A - 6-o-alpha-d-galactopyranosyl-l-ascorbic acid or its salt, production and use thereof - Google Patents

Abstract

PURPOSE:To obtain a new safe compound, excellent in preservation stability, capable of sufficiently exhibiting the activity of L-ascorbic acid in the living body, suitable for blending as a vitamin C nutrient supplement, a stabilizer, etc., in a food, drink, a nosotropic agent for susceptibility, etc., and producible in good yield. CONSTITUTION:The objective 6-O-(alpha-D-galactopyranosyl-L-ascorbic acid of the formula or its salt. This compound is obtained by reacting L-ascorbic acid or its salt with a solution comprising a compound containing a-galactosyl group (e.g. melibiose or phenyl-alpha-galactoside) with an alpha-galactosidase, preferably at 40-70 deg.C temperature and pH 3.5-8. Furthermore, an alpha-galactosidase produced by Candida guilliermondii H-404 (FERM P-11026) is preferred as the alpha- galactosidase from the viewpoint of the yield.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a highly stable L-ascorbic acid sugar derivative, that is, 6-O-α-D-galactopyranosyl-L-ascorbic acid or a salt thereof, a process for producing the same and uses thereof. It is about.

[0002]

2. Description of the Related Art L-ascorbic acid (vitamin C) is synthesized from L-glonolactone in the liver of mammals, but in humans, monkeys and guinea pigs, it is necessary to take it in as an essential vitamin from the outside because the synthase is deficient. There is. In vivo, L-ascorbic acid is involved in the hydroxylation of proline and lysine necessary for the synthesis of collagen, which is the main component of biological connective tissue, and reduces Fe ³⁺ of cytochrome C to Fe ²⁺ . It is also known to be involved in redox reactions such as oxidization, and also to prevent damage by radicals, immunopotentiation by increasing leukocytes, antiarteriosclerosis by suppressing cholesterol production, and antiallergic effect by antihistamine. Plays an important role in maintaining and promoting health. Scurvy has long been known as a deficiency of L-ascorbic acid, and weakens the skin,
Presents with petechiae, plaques, gingiva or submedullary hemorrhage.
To prevent this, 50 mg /
It is considered necessary to take daily L-ascorbic acid. As described above, L-ascorbic acid is used not only as a vitamin C enhancer as an essential nutrient supplier, but also as a sour agent, a reducing agent, an antioxidant, a bleaching agent due to its chemical structure and physiological activity. As a stabilizer, various chemical reaction bases, foods and drinks, etc., and also as a preventive agent, therapeutic agent or anti-susceptive agent for viral diseases, bacterial diseases, malignant tumors, susceptibility diseases such as diabetes, and further UV rays. It has an extremely wide range of applications including cosmetics such as absorbents, skin-beautifying agents and skin-whitening agents that suppress melanin production.
However, this L-ascorbic acid is a very unstable substance, and it is known that it is easily decomposed or modified by heat, light, oxygen, metal ions or the like. There have been many attempts to stabilize such unstable L-ascorbic acid, and several derivatives have been developed. For example, ascorbic acid phosphoric acid ester (JP-A-57-1
40789), glycosyl ascorbic acid (Japanese Patent Publication Sho 5)
No. 8-5920), ascorbic acid fatty acid ester (JP-A-62-84072), L-ascorbic acid derivative and a method for producing the same (JP-A No. 2-311490), and the like.

[0003]

As mentioned above, various ascorbic acid derivatives have been developed. Among them, the sugar derivatives are excellent in stability and are expected to be put to practical use. The yield at the time of reaction was low and there was a problem to be solved. The present invention provides an L-ascorbic acid sugar derivative or a salt thereof which is safe, has excellent storage stability, and can sufficiently exhibit the physiological activity of L-ascorbic acid in vivo, and a process for producing the same at a high yield. The present invention further aims to provide foods and drinks, anti-susceptive disease agents, and cosmetics containing the L-ascorbic acid sugar derivative as an active ingredient.

[0004]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that a mixture of L-ascorbic acid or a salt thereof and a galactose donor is treated with α-galactosidase. Operate under mild conditions, α
-By utilizing the transglycosylation reaction of galactosidase, it was found that the target L-ascorbic acid sugar derivative can be synthesized in good yield, and the present invention has been completed.

That is, the present invention is represented by the following formula 6
-O-α-D-galactopyranosyl-L-ascorbic acid or a salt thereof, and α-galactosidase are allowed to act on a solution of L-ascorbic acid or a salt thereof and a compound containing an α-galactosyl group. 6-O-
A method for producing α-D-galactopyranosyl-L-ascorbic acid or a salt thereof, and 6-O-α-D-galactopyranosyl-L-ascorbic acid or a salt thereof as an active ingredient. Food and drink, anti-susceptive agent,
And cosmetics are the main points.

[0006]

[Chemical 2]

The present invention will be described in detail below. The 6-O-α-D-galactopyranosyl-L-ascorbic acid of the present invention has a structure represented by the above formula 2, and is L-ascorbic acid which is a component used in medicines, foods, cosmetics and the like. It is composed of galactose and may be a salt such as sodium salt, potassium salt or lithium salt when used as an active ingredient of the food, drink, anti-susceptive disease agent and cosmetics of the present invention.

The α-galactosidase used in the present invention is reacted with a solution containing a galactosyl group-containing compound and L-ascorbic acid to give 6-glycol by a transglycosylation reaction.
Any source can be used as long as it synthesizes O-α-D-galactopyranosyl-L-ascorbic acid, and the origin and type are not limited. For example, Pycnopor
us cinnabarinus), Streptococcus bovis (St
reptococcus bovis), Diplococcus pneumoniae, Mortierella vinacea, Aspergillusnigar, Escheri Escheri
chia coli), Pseudomonas fluorescens (Pse
udomonas fluorescens), microorganisms such as Candida guilliermondii, Visia sativa, green coffee beans (Gr)
α-galactosidase produced by plants such as een coffee bean) can be used, but α-galactosidase produced by Candida guilliermondii is preferable from the viewpoint of yield.

Candida guilliermond used in the present invention
ii is ATCC6260, ATCC9058, AT
CC9390, ATCC14242, ATCC2201
7, ATCC22948, ATCC22949, ATC
C22995, ATCC26860, ATCC2887
3, ATCC32542, ATCC34134, ATC
C38290, ATCC42050, ATCC5682
2, IFO0437, IFO0454, IFO067
9, IFO0837, IFO0838, IFO191
3, IFO1972, H-404
MP-11026), but Candida guilliermondii H-404 in terms of reaction yield.
(Micromachinery Research Institute, FERM P-11026) is excellent.

As a method for producing α-galactosidase from these microorganisms, liquid culture or solid culture is usually used. In the case of liquid culture, the culture supernatant can be used as it is, and in the case of solid culture, the extract can be used as it is as an enzyme preparation. In some cases, the bacterial cells can be used as they are as an enzyme agent. In addition, an enzyme produced by a known method can be used if necessary. It is also possible to immobilize these enzymes or bacterial cells that produce the enzymes and use them in a continuous reaction or a batch reaction repeatedly.
Further, if necessary, L-ascorbic acid and a compound containing an α-galactosyl group are allowed to coexist during growth of a microorganism having an α-galactosyl group transfer reaction ability to give 6-O-α-
It is also possible to generate D-galactopyranosyl-L-ascorbic acid.

In the present invention, L-ascorbic acid includes
Not only free L-ascorbic acid, but also L-ascorbic acid metal salts such as alkali metal salts or alkaline earth metal salts thereof, or a mixture thereof. The compound containing an α-galactosyl group used as a substrate for α-galactosidase may be a natural product, an organic synthesis product or an enzymatic reaction product, regardless of its origin. Examples of natural products include melibiose, manninotriose, raffinose, stachyose, velbaucose, galactomannans such as guar gum and locust bean gum, and mixtures thereof, and organic compounds such as phenyl-α-galactoside and paranitrophenyl-. α-galactoside, ortho-nitrophenyl-α-galactoside, metanitrophenyl-α-galactoside, etc., as the enzymatic reaction product, melibiose, an enzymatic decomposition product of galactomannan such as guar gum or locust bean gum, or high-concentration galactose. An α-galacto-oligosaccharide composition produced by allowing α-galactosidase to act on a lactose hydrolyzate is used.

In the present invention, the concentration of L-ascorbic acid during the reaction is preferably 1 to 60% (W / V), preferably 5 to 40% (W / V), and 10 to 30%.
(W / V) is more preferable. The appropriate concentration of melibiose is 1-80% (W / V), 5-60%
(W / V) is preferable, and 10-40% (W / V) is more preferable. The concentration of lactose is 1 to 80% (W /
V) is suitable, 5-60% (W / V) is preferable,
10 to 40% (W / V) is more preferable.

The pH during the reaction is preferably 3.5 to 8.0, but the pH may be selected according to the enzyme used.

Next, the reaction temperature is suitably in the range of 10 to 90 ° C., preferably in the range of 30 to 80 ° C., and 40 to 70 ° C.
Is more preferable.

The reaction time is not particularly limited, but a time may be selected so that the production amount of the L-ascorbic acid sugar derivative is maximized.

The L-ascorbic acid sugar derivative produced by such a method should be separated by a usual separation means such as membrane separation, ion exchange column chromatography, activated carbon column chromatography, liquid chromatography, silica gel column chromatography and the like. You can At this time, the separated unreacted L-ascorbic acid and the compound containing the α-galactosyl group can be reused as a raw material for the transglycosylation reaction.

6-O-α-D-galactopyranosyl-L-ascorbic acid, which is an active ingredient of the food, drink, anti-susceptive diseases and cosmetics of the present invention, is a stable and safe vitamin C enhancer. Not only as taste improver, sour agent, stabilizer, quality improver, antioxidant, UV absorber, etc.
It can be advantageously used by blending it in 0.001% by weight or more according to the purpose thereof in foods and drinks, favorite foods, feeds, prophylactic agents and therapeutic agents for susceptive diseases, cosmetics such as skin beautifying agents and skin lightening agents.

Further, 6-O-α-D-galactopyranosyl-L-ascorbic acid is well harmonized with various substances having tastes such as sourness, saltiness, astringency, umami and bitterness,
As it has high acid resistance and heat resistance, it can be used for ordinary ordinary food and drinks, favorite foods such as synthetic liquor, fruit liquor, Western liquor, liquor such as chuhai, tea, juice, carbonated drink, lactic acid drink, lactic acid bacterium drink, sports drink, Instant beverages such as soft drinks, instant juice, instant tea, instant soup, soy sauce, powdered soy sauce, miso, powdered miso, moromi, sauce, ketchup,
Mayonnaise, dressing, lemon juice, vinegar, three cups vinegar, sushi vinegar, powdered sushi vinegar, mirin, mirin-style seasoning, tempura sauce, noodle sauce, Chinese sauce, dumpling sauce, roasted meat sauce, curry roux, stew, soup sauce, Dashi stock, various seasonings such as complex seasonings, salad oil, oil and fat foods such as margarine, syrup pickled fruits, syrups such as ice honey, butter cream, custard cream,
Spread pastes such as peanut paste, flower paste, fruit paste, jam, marmalade, syrup pickled fruits, processed fruits and vegetables,
Rice crackers, hail, rice cakes, steamed buns, uiro, rice cakes, bean paste, yokan, jelly, castella, hard candy and other Japanese sweets, bread, cookies, crackers, pies, biscuits,
Pudding, cakes, cream puffs, waffles, donuts, chocolate, chewing gum, caramel, candies, snacks and other Western confectionery, ice cream, sherbet and other ice confectionery, bread, cooked rice and other cereals, pickles, meat products , Fish meat products, boiled foods, prepared foods, milk drinks, dairy products, bottled products such as meat, fish meat, fruits, vegetables, canned products, etc., vitamin C enhancer, taste improver, acidulant, quality improver , It can be advantageously used as a stabilizer, an antioxidant and the like. Also,
For livestock, poultry, bees, silkworms, feeds and other feeds for domestic animals such as fish, for the purpose of vitamin C enhancer, taste improver, quality improver, stabilizer, antioxidant, palatability improver, etc. It can be blended.

In addition, troches, liver oil drops, complex vitamins, tube feedings, oral medicines, injections, toothpaste paste, mouthwash, mouthwash, lipstick, eye shadow,
Advantageously blended in various solids such as lotions, foundations, emulsions, hand creams, sunscreen creams, face soaps, shampoos, rinses, hair styling agents, pastes, liquids, cosmetics such as skin-care agents and whitening agents. Can be implemented. Further, it can be used as an ultraviolet absorber, a deterioration preventing agent, etc. by blending it with a plastic product or the like.

6-O-α-D-galactopyranosyl-L
-A disease that is prevented or treated by ascorbic acid is a susceptible disease in the present invention, and includes, for example, viral disease, bacterial disease, traumatic disease, immune disease, allergic disease, diabetes, cataract, malignant tumor, arteriosclerosis, etc. It may be. The shape of the prophylactic or therapeutic agent for 6-O-α-D-galactopyranosyl-L-ascorbic acid-sensitive diseases can be freely selected according to the purpose. For example, sprays, eye drops, nasal drops, mouthwashes, injections and other liquids,
Ointment, dusting agent, paste, powder, granule, capsule,
Examples include solid agents such as tablets and suppositories. When necessary, one or two other components such as therapeutic agents, physiologically active substances, antibiotics, adjuvants, bulking agents, stabilizers, coloring agents, flavoring agents, excipients, etc. It is also possible to use in combination with the above.

6-O-α-D-galactopyranosyl-L
The dose of ascorbic acid can be appropriately adjusted depending on the content, administration route, administration frequency and the like. The amount of 0.0005 to 100 g per day for an adult is good, preferably 0.0
The range of 05 to 50 g, more preferably 0.01 to 10 g is suitable.

6-O-α-D-galactopyranosyl-L
As a method of incorporating ascorbic acid, a known method such as mixing, dissolving, dipping, spraying, coating, spraying or pouring can be appropriately selected in the steps until the products are completed. In addition, 6-O-α-D-galactopyranosyl-
When L-ascorbic acid is a free acid, it is reacted with an aqueous solution of metal hydroxide, metal carbonate or the like, if necessary, to give 6-O-α-D-galactopyranosyl-L-ascorbic acid. It is also possible to use salt as a salt to adjust the pH and to prepare a substance having both a vitamin C action and a mineral, which can be used as a nutrient enhancer, a chemical, or the like.

[0023]

EXAMPLES Next, the present invention will be specifically described by way of examples. Example 1 Sodium L-ascorbate (Ishizu Pharmaceutical Special Grade Reagent) 1
0 g and 20 g of melibiose (manufactured by Sigma) were dissolved in distilled water to make 100 ml, and then the pH was adjusted to 4.5. To this, 1900 units of solution type α-galactosidase derived from Candida guilliermondii H-404 (Microtechnology Research Institute, FERMP-11026) was added and reacted at 50 ° C. for 5.5 hours. Solution type α-galactosidase is Ca
ndida guilliermondii H-404
MP-11026) with 1% melibiose, 3% polypeptone, 0.5% yeast extract, 0.05% K ₂ HPO.
_{4, 0.01% MgSO 4 · 7H} 2 O, 0.01% KC
60 ml of sterile liquid medium (pH 7.0) containing 1
By shaking culture at 30 ° C. for 4 days in a ml Sakaguchi flask, 0.8 unit / ml in the supernatant and 35 units / g in wet cells could be obtained. The activity was measured with 10 mM para-nitrophenyl-α-galactoside.
0.05 ml of α-galactosidase solution in 2 ml and 0.2 ml of 40 mM buffer (pH is in accordance with the optimum pH of the enzyme)
1 and then reacted at 40 ° C. for 10 minutes, 0.2 M Na
_The reaction was stopped by adding 0.5 ml of ₂ CO ₃ and the amount of liberated para-nitrophenol was measured by measuring the absorbance at 400 nm. 1 unit is 1 μm per minute
It was defined as the amount of enzyme that produces ole paranitrophenol.

The obtained reaction product was designated as Bio-Rad (BIO-RAD
) Column high-performance liquid chromatography column Aminex ion exclusion (AMINEX ION EXC
LUSION) HPX-87H. At this time,
0.01N H ₂ SO ₄ was used as the eluent, and the flow rate was 0.
The reaction was eluted at 6 ml / min. The detection at that time was performed with UV ₂₄₅ . As a result, 6.1 g of L-
A sugar derivative of ascorbic acid was produced. The reaction yield at this time was 30.5%.

Next, 50 ml of the above reaction product was applied to a 450 ml activated carbon column (for chromatography by Wako Pure Chemical Industries) and eluted with water at 10 ml / min. At this time, L-
Since ascorbic acid and L-ascorbic acid sugar derivative were eluted in this order, only the eluted L-ascorbic acid sugar derivative was collected and concentrated. The concentrate was added to 5 ml of anion exchange resin Dowex 2 × 8 column (Cl ⁻
Type), 100 ml of distilled water was passed through at a flow rate of 1 ml / min, and then 100 ml of 0.04 M NaCl was added.
After passing through, the target L-ascorbic acid sugar derivative was eluted. This fraction was desalted with an electrodialyzer (Micro Asylizer, manufactured by Asahi Kasei Corp.), freeze-dried to give 2.1 g of L.
A sugar derivative of ascorbic acid was obtained. The purification yield at this time was 68.8%.

¹³ C-NMR analysis was performed on the obtained L-ascorbic acid sugar derivative, and the following results were obtained. ¹³ C-NMR (σ value ppm) C-1 101.5 of galactose C-2 71.2 of galactose C-3 71.3 of galactose C-4 72.1 of galactose C-5 72.3 galactose of galactose C-6 64.0 L-ascorbic acid C-1 116.3 L-ascorbic acid C-2 180.1 L-ascorbic acid C-3 176.9 L-ascorbic acid C-4 81. 1 L-ascorbic acid C-5 73.9 L-ascorbic acid C-6 70.6

Further, the obtained L-ascorbic acid sugar derivative is hydrolyzed with α-galactosidase derived from Aspergillus niger to give L
-Ascorbic acid and galactose produced.

From the above results, it was revealed that the structure of the L-ascorbic acid sugar derivative obtained above was 6-O-α-D-galactopyranosyl-L-ascorbic acid.

Example 2 20 g of lactose (Ishizu Pharmaceutical Special Grade Reagent) was dissolved in distilled water, and p
After adjusting to H6, the volume was adjusted to 80 ml. To this solution, 800 units of β-galactosidase (manufactured by Sigma) derived from Aspergillus oryzae was added and reacted at 40 ° C for 10 hours. After completion of the reaction, the enzyme was inactivated by boiling to obtain a lactose hydrolyzate. This to Cand
ida guilliermondii H-404 (Microtech Lab.
P-11026) -derived α-galactosidase 1
Add 890 units, react at 50 ° C for 16 hours, and α-
A galactooligosaccharide composition was synthesized. Then, 15 g of sodium L-ascorbate (Ishizu Pharmaceutical Special Grade Reagent) was added and dissolved to make 100 ml, and the mixture was reacted at 50 ° C. for 5.5 hours.

When analyzed by high performance liquid chromatography in the same manner as in Example 1, 9.7 g of L-ascorbic acid sugar derivative was formed. The reaction yield at this time is 3
It was 2.3%.

Next, the L-ascorbic acid derivative was purified in the same manner as in Example 1 to obtain 6.7 g of the L-ascorbic acid sugar derivative. The purification yield at this time was 69.0%.

Example 3 Raffinose pentahydrate 1.7 g and L-ascorbic acid 1
pH 5.5 containing 90 units of commercially available Mortierella vinase-derived α-galactosidase (Seikagaku Corporation)
10 ml of the above solution was prepared and reacted at 50 ° C. for 8 hours.
The reaction solution was purified according to the method of Example 1, concentrated and powdered to obtain 0.13 g of a powdery product. Next, when the structure was analyzed in the same manner as in Example 1, it was 6-O-α-D-galactopyranosyl-L-ascorbic acid.

Example 4 Stability test of 6-O-α-D-galactopyranosyl-L-ascorbic acid Using the following components, a soft drink containing grapefruit juice was obtained according to a conventional method. Ingredients wt% Grapefruit juice 20.0 Sugar 12.0 Citric acid 1.0 6-O-α-D-galactopyranosyl-L-ascorbic acid 0.1 obtained in Example 1 0.1 Prepared to 100% water.

The obtained soft drink was stored at 40 ° C. for 120 days, and a stability test was conducted. That is, the vitamin C remaining in the soft drink water was analyzed by high performance liquid chromatography (LC-2 manufactured by Shimadzu Corporation, column: Bio-Rad AMI).
(NEX HPX-87H, flow rate: 0.6 ml / min, solvent solution: 0.01 MH ₂ SO ₄ , detection wavelength: 245 nm)
Was quantified at. The results are shown in Table 1. The numbers in Table 1 show the residual rate of vitamin C.

[0035]

[Table 1]

Next, the vitamin C activity test of the soft drink water obtained in Example 4 was conducted. That is, non-fat dry milk, non-fat soybeans, and various basic vitamins other than vitamin C were used as a basic feed, and Hartley male guinea pigs (body weight approximately 250
G, 10 fish) was bred for 20 days, and the resulting soft drink (6-O-α-D-galactopyranosyl-L-ascorbic acid concentration: 1.7 mg / ml) was orally administered 1 ml each day. It was administered and the change in body weight was measured. As a control group, 1 ml of distilled water was orally administered daily. The test results are shown in Table 2. The numbers in Table 2 show changes in the average body weight of guinea pigs.

[0037]

[Table 2]

Comparative Example 1 The procedure of Example 4 was repeated except that 2-O-glycosyl-L-ascorbic acid was used instead of 6-O-α-D-galactopyranosyl-L-ascorbic acid. Drinking water was obtained and a stability test was conducted as in Example 4. The results of the stability test are shown in Table 3.

Comparative Example 2 The procedure of Example 4 was repeated, except that 6-O-stearyl-L-ascorbic acid was used instead of 6-O-α-D-galactopyranosyl-L-ascorbic acid. Drinking water was obtained and a stability test was conducted as in Example 4. The results of the stability test are shown in Table 3.

Comparative Example 3 A soft drink was prepared in the same manner as in Example 4 except that L-ascorbic acid was used instead of 6-O-α-D-galactopyranosyl-L-ascorbic acid. A stability test was conducted as in Example 4. The results of the stability test are shown in Table 3.

[0041]

[Table 3]

As is clear from Table 3, the soft drink containing L-ascorbic acid (Comparative Example 3) and 6-O-stearyl-L-ascorbic acid as components (Comparative Example 2) is very unstable, The amount of added vitamin C decreased with time. Further, the soft drink containing 2-O-glycosyl-L-ascorbic acid as a component (Comparative Example 1) is relatively stable, but the stability is not so excellent as compared with the soft drink of the present invention. It was On the other hand, the soft drink (Example 4) containing 6-O-α-D-galactopyranosyl-L-ascorbic acid of the present invention as a component has a temperature of 40 ° C.
It was very stable in the long term.

[0043]

[Table 4]

As is clear from Table 4, in the control group, basal feed was given but vitamin C was deficient, so almost no weight gain was observed after the 7th day, and 3 out of 10 fish died after the 14th day. did. On the other hand, the group to which the soft drink containing 6-O-α-D-galactopyranosyl-L-ascorbic acid as a component (Example 4) was administered was a soft drink containing L-ascorbic acid as a component. Similar to the group to which (Comparative Example 3) was administered, the average daily weight gain was 4 to 5 g, and no abnormality was observed during the administration period. That is, the soft drink water (Example 4) containing 6-O-α-D-galactopyranosyl-L-ascorbic acid as a component of the present invention exhibits the same vitamin C activity as L-ascorbic acid in vivo. It was a thing.

Example 5 6-O-α-D-galactopyranosyl-L-ascorbic acid antioxidative activity test 6-O-α-D-galactopyranosyl-L-obtained in Example 1 1 g of ascorbic acid was dissolved in 10 ml of distilled water, and the whole amount was sprayed on 100 g of fried beans and dried to obtain a bean confectionery.

The obtained bean confectionery was stored at 40 ° C., 5 g was weighed out with time, and the solvent (chloroform: glacial acetic acid = 2:
After the extraction in 3), potassium iodide was added, the liberated iodine was titrated, and the increase in the peroxide value was measured to carry out the antioxidant capacity test. As a control group, an unsprayed bean confectionery was obtained, and the antioxidant ability test was conducted in the same manner. The test results are shown in Table 5. The numbers in Table 5 show the peroxide value.

Comparative Example 4 The procedure of Example 5 was repeated except that 2-O-glucosyl-L-ascorbic acid was used instead of 6-O-α-D-galactopyranosyl-L-ascorbic acid. Bean confectionery was obtained, and the antioxidant ability test was conducted in the same manner as in Example 5. The results of the antioxidant capacity test are shown in Table 5.

Comparative Example 5 A bean confectionery product was obtained in the same manner as in Example 5, except that BHA (butylhydroxyanisole) was used instead of 6-O-α-D-galactopyranosyl-L-ascorbic acid. The antioxidant ability test was conducted in the same manner as in Example 5. The results of the antioxidant capacity test are shown in Table 5.

[0049]

[Table 5]

As is apparent from Table 5, the confectionery prepared by spraying 2-O-glucosyl-L-ascorbic acid in which the hydroxyl group at the 2-position of L-ascorbic acid, which is considered to be an antioxidant active center, was sprayed (Comparative Example 4). The peroxide value of () showed almost the same increase as that of the control group. On the other hand, in the bean candy (Example 5) sprayed with 6-O-α-D-galactopyranosyl-L-ascorbic acid of the present invention, the increase in peroxide value was suppressed to about 60% of the control group. Compared with the case of spraying BHA known as an antioxidant (Comparative Example 5), the increase was only about 30%, showing a high antioxidant activity. That is, the bean confectionery (Example 4) sprayed with 6-O-α-D-galactopyranosyl-L-ascorbic acid of the present invention exhibits high antioxidant activity. Further, the bean confectionery obtained in Example 5 had stability and in vivo vitamin C activity similar to those in Example 4.

Example 6 Method for producing soft drink containing grapefruit juice Using these ingredients, a soft drink containing grapefruit juice was obtained according to a conventional method.

Example 7 Preparation method of chewing gum Using these ingredients, chewing gum was obtained according to a conventional method.

Example 8 Method for producing sweet bread Using these ingredients, confectionery bread was obtained according to a conventional method.

Example 9 Method for producing mayonnaise Using these ingredients, mayonnaise was obtained according to a conventional method.

Example 10 Preparation Method of Eye Drops and Eye Irritation Test Using 6-O-α-D-galactopyranosyl-L-ascorbic acid obtained in Example 1 and the following formulation, according to a conventional method. An eye drop was prepared. Next, a panel of 50 people evaluated the eye irritation of the eye drop having the above formulation. As a result, most did not complain of irritation.

Example 11 Preparation method of toothpaste and storage colorability test

Comparative Example 6 Instead of 6-O-α-D-galactopyranosyl-L-ascorbic acid (Example 11), a toothpaste containing the same amount of L-ascorbic acid and a control paste containing no toothpaste were added. Toothpaste was prepared. 30 toothpastes of Example 11 and Comparative Example 6 respectively
It was stored at 6 ° C. for 6 months, at 40 ° C. for 3 months, and at 50 ° C. for 3 months, and the coloring degree was judged according to the following criteria. -: No coloring +: Minor coloring ++; Heavy coloring

The results are shown in Table 6.

[0059]

[Table 6]

As is clear from Table 6, the toothpaste containing L-ascorbic acid (Comparative Example 6) was markedly colored, but 6-O-α-D-galactopyranosyl-L-ascorbic acid was added. The toothpaste prepared (Example 11) showed no coloration.

Next, the toothpaste of Example 11 and Comparative Example 6 was used by 30 patients with degree 1 alveolar pyorrhea for 6 months, and redness of the gingiva, disappearance of pus, improvement of the blind sac, and improvement of tooth movement. And the effectiveness was judged using the improvement of gingiva as an index. During the test, the patient was asked to brush his teeth twice a day in the morning and evening. The judgment was made every two months according to the following criteria. 5 points: 5 items in the index were improved. 4 points: 4 items in the index were improved. 3 points: 3 items in the above index were improved. 2 points: Two items in the above index were improved. 1 point: One item in the index was improved. 0 point: No improvement or deterioration. Table 7 shows the total score of each dentifrice use group divided by the number of patients on each determination day.

[0062]

[Table 7]

6-O-α-D-galactopyranosyl-L
-No case was worsened by the use of ascorbic acid-containing toothpaste (Example 10), and abnormalities in the oral cavity due to its use,
There was no sense of discomfort.

Example 13 Method for Producing Oral Pasta Using the 6-O-α-D-galactopyranosyl-L-ascorbic acid obtained in Example 1 and the following formulation, an oral pasta was prepared according to a conventional method. Obtained. Ingredients Weight g Ethanol 10.0 Squalene 20.0 Precipitable silica 5.0 Polyoxyethylene hydrogenated castor oil 0.1 Sodium lauryl sulfate 0.2 EDTA calcium 0.1 Perfume 0.6 Sorbitan monooleate 1.0 Glycyrrhizin Acid 0.1 Saccharin sodium 0.6 6-O-α-D-galactopyranosyl-L-ascorbic acid 0.5 The oral pasta obtained in Example 13 was as stable and improved as Example 11. Showed the effect.

Example 14 Method for Producing Oral Lozenge Using the 6-O-α-D-galactopyranosyl-L-ascorbic acid obtained in Example 1 and the following formulation, the ingredients were mixed well according to a conventional method. A troche for oral cavity was obtained by direct compression. Ingredients Weight g Lactose 97.0 Polyoxyethylene monostearate 0.2 Sodium lauryl sulfate 0.05 Chlorhexidine gluconate 0.02 Stevia extract 0.2 Perfume 0.1 Hydroxyethylcellulose 1.93 6-O-α- D-Galactopyranosyl-L-ascorbic acid 0.5 The oral troche obtained in Example 14 showed the same degree of stability and improvement effect as Example 11.

Example 15 Preparation Method of Mouthwash Using the 6-O-α-D-galactopyranosyl-L-ascorbic acid obtained in Example 1 and the following formulation, the mixture was mixed well according to a conventional method to prepare a mouse. I got a wash. Ingredients g Solbit 10.0 Ethanol 5.0 Polyoxyethylene hydrogenated castor oil 0.1 Sucrose monopalmitate 0.2 Sodium lauryl sulfate 0.05 Sodium saccharin 0.2 Sodium hydrogencarbonate 0.05 EDTA calcium 0.05 Perfume 0.6 Distilled water 83.65 6-O-α-D-galactopyranosyl-L-ascorbic acid 0.1 The mouthwash obtained in Example 15 was as stable and improved as Example 11. showed that.

Example 16 Method for producing creamy cosmetic product A creamy cosmetic product was obtained by a conventional method using the components of the following formulation. Ingredient Weight g Squalane 25.0 Wax 5.0 Olive oil 3.0 Xanthan gum 1.0 Monoammonium glycyrrhizinate 0.3 Methylparaben 0.2 Distilled water 61.5 Fragrance Suitable amount 6-O-α-D-galactopyranosyl- L-ascorbic acid 4.0

The above cream was applied to the face of 20 panelists (female) suffering from dark blacks, stains and freckles 2 times a day.
The panelist himself evaluated the effect on three levels: effective, moderately effective, and ineffective. Further, instead of 6-O-α-D-galactopyranosyl-L-ascorbic acid, the same amount of L-ascorbic acid was added,
A creamy cosmetic was obtained and tested in the same manner, and the results are shown in Table 8.

[0069]

[Table 8]

Further, the above cream was stored at 40 ° C. for 6 months and examined for appearance changes and odor. Table 9 shows the experimental results.

[0071]

[Table 9]

As is clear from Tables 8 and 9, the skin cosmetics of the present invention were found to be effective in terms of storage stability, whitening effect and rough skin improving effect.

[0073]

INDUSTRIAL APPLICABILITY The 6-O-α-D-galactopyranosyl-L-ascorbic acid or its salt, which is the L-ascorbic acid sugar derivative of the present invention, can be produced in good yield at low cost and is stable. It has excellent properties, is non-toxic because it is easily hydrolyzed in vivo, and exhibits sufficient vitamin C activity in vivo, and is a stabilizer, quality improver, antioxidant, bioactive agent, ultraviolet absorber, etc. As beverages, processed foods, seasonings,
It can be advantageously used by including it in foods and drinks such as favorite foods, preventive agents and therapeutic agents for susceptive diseases, and also cosmetics such as skin beautifying agents and skin lightening agents.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Office reference number FI technical display location A61K 7/48 9051-4C 31/70 ABA ADK 8314-4C C09K 15/06 C12N 9/40 7823- 4B C12P 19/58 7432-4B (72) Inventor Satoshi Kawanaka 24 Uji Kozakura, Uji-shi, Kyoto Prefecture (72) Inventor Munehiko Muho 24 24 Uji Kozakura, Uji-shi, Kyoto Prefecture Inventor Hiroshi Nakajima Uji Kozakura, Uji-shi, Kyoto Prefecture twenty four

Claims (5)

[Claims] 1. 6-O-α-D-galactopyranosyl-L-ascorbic acid represented by the following formula or a salt thereof. [Chemical 1] 2. L-ascorbic acid or a salt thereof, and α
-Α-in a solution containing a compound containing a galactosyl group
A method for producing 6-O-α-D-galactopyranosyl-L-ascorbic acid or a salt thereof according to claim 1, wherein galactosidase is allowed to act. 3. A food or drink comprising the 6-O-α-D-galactopyranosyl-L-ascorbic acid or the salt thereof according to claim 1 as an active ingredient. 4. An anti-susceptive disease agent comprising the 6-O-α-D-galactopyranosyl-L-ascorbic acid or the salt thereof according to claim 1 as an active ingredient. 5. A cosmetic comprising the 6-O-α-D-galactopyranosyl-L-ascorbic acid or salt thereof according to claim 1 as an active ingredient.