JPH05310586A - Active oxygen eliminating agent prepared from rice bran and wheat bran - Google Patents

Abstract

PURPOSE:To provide an active oxygen eliminating agent producible at a low cost, having excellent effect to eliminate active oxygen causing various disorders and safely usable in wide field such as pharmaceuticals, foods and cosmetics. CONSTITUTION:Rice bran or wheat bran is extracted with water or an organic solvent and the extract is used as the active oxygen eliminating agent or as a component of the agent.

Description

Detailed Description of the Invention

[0001]

INDUSTRIAL APPLICABILITY The present invention is a safe, inexpensive, and safe scavenging effect of active oxygen causing various obstacles, which is obtained by using a water extract or an organic solvent extract from rice bran or bran as it is or containing them. , An active oxygen scavenger that can be used in a wide range of fields such as pharmaceuticals, foods, and cosmetics.

[0002]

2. Description of the Related Art When human beings maintain a healthy body, SOD which is an active oxygen scavenging enzyme in the living body and active oxygen in the body
(Superoxide dismutase) is always balanced and the concentration of active oxygen is kept almost constant. However, currently, SOD production is decreased due to imbalance of dietary habits, excessive stress, and aging, and on the other hand, active oxygen is increased due to smoking, air pollution and the like.

As a result, active oxygen is excessively present in the living body, causing various tissue disorders. Particularly in the elderly, the SOD activity decreases and the active oxygen concentration increases, causing disorders such as rheumatoid arthritis and Pechet's disease. In addition, lipid peroxides generated by active oxygen, myocardial infarction, stroke, cataract, spots, freckles, wrinkles,
It is a leading cause of modern diseases such as diabetes, arteriosclerosis, stiff shoulders, and coldness.

In addition, even if not aged, active oxygen is particularly likely to be generated at a site such as the skin which is directly stimulated by environmental factors such as ultraviolet rays, so that the melanin pigment increases with the increase of the active oxygen concentration. It is easy to cause problems such as the generation of blemishes, stains, and wrinkles.

Therefore, SODs that eliminate excess active oxygen, which causes various disorders as described above, have attracted attention, and in order to prevent or treat these disorders, SODs are used as medicines, cosmetics and foods. Attempts have been made to add and use.

However, since SOD is unstable to heat, deactivates by oral administration, and is extremely expensive, elimination of active oxygen by SOD has not been successful yet.

[0007] In view of the above circumstances, studies on active oxygen scavengers (including antioxidants that act similarly to SOD enzymes) have been conducted, and active oxygen scavengers using crude drug extract and the like have also been developed. At present, not only is it expensive due to a special raw material, but it is also difficult to supply a stable material.

[0008]

Since various obstacles caused by active oxygen have been recognized as described above, various studies have been actively conducted to eliminate active oxygen in the living body. In addition, nowadays, it is desired to live in a healthier and older life with the aging society. On the other hand, active oxygen scavengers are also attracting attention from the viewpoint of beauty. Therefore, there is a demand for the development of an active oxygen scavenger that is safe and inexpensive for the human body, has an excellent effect of scavenging active oxygen that causes various disorders, and that can be easily manufactured and can be stably supplied.

[0009]

[Means for Solving the Problems] From the viewpoint of animal and plant harmony, the present inventors have conducted research on various plant components centering on rice, which is a staple food. In the process, it was found that by-products of cereals such as rice bran and bran had an extremely excellent active oxygen scavenging effect which could not be predicted until now, and the present invention was completed.

That is, the present invention is an active oxygen scavenger characterized by containing a water extract or an organic solvent extract from rice bran or bran as it is or containing the same. By extraction (including acid and alkali extraction) or extraction with an organic solvent such as alcohol, a very excellent active oxygen scavenger exhibiting the above effects can be obtained simply, inexpensively and completely safely.
Examples of the raw material of the present invention include bran such as rice, barley, fluff, and loin, and bran such as wheat and rye. However, flour of the skin portion such as pericarp and seed coat of all grains can be used.

In extracting water, rice bran or bran is watered. The amount of water added is efficiently extracted in an amount of 2 to 5 times the amount of the sample, and may be appropriately selected depending on the yield, workability, end use purpose and the like. After that, the mixture is heated and the extraction is completed at the time when it reaches a boiling state. After the extraction is completed, squeezing and filtration are carried out depending on the purpose of use to obtain a clear extract. Note that hot water may be added from the beginning to perform extraction.

Although the active ingredient in the extract has not been clarified, it has been confirmed that this unknown active ingredient is stable to heat. Therefore, the extraction temperature during water extraction is effectively high. If it is left at a low temperature for a long time, it can be sufficiently extracted. However, at a low temperature of 40 ° C. or lower, it is necessary to make the pH acidic or alkaline or to add a preservative. The extraction time may be short in the case of boiling extraction, but if the temperature is lower than that, several hours to one day is required. If the temperature is low, it takes several days to one month. However, even in this case, it is more effective to heat at the end as much as possible.

Since the active ingredient in the extract is stable to acid and alkali, acid extraction or alkali extraction is also effective. Further, in the case of water extraction, it is efficient to carry out a pretreatment by reacting an enzyme (lipase, cellulase, etc.) that acts on the bran and bran to perform extraction. This is probably because the pretreatment makes it easier for the active ingredient to be extracted.

Furthermore, it has been found that an extract having this effect can be extracted even by extraction with an organic solvent. This is extremely effective in promoting the elucidation of the active ingredient, and also in the usage of extracting the active ingredient into a concrete or blending it with a substance insoluble in water. It is desirable to use an organic solvent such as alcohol that is safe for administration to the human body.

It is more effective to extract the active ingredient from the bran or bran of the present invention by organic solvent extraction or water extraction as described above and further solvent extraction of the active ingredient in the extract. However, this is thought to be because a more concentrated state can be obtained, and the same effect can be obtained by concentrating. Furthermore, even if the extract was combined with fermentation such as alcoholic fermentation and lactic acid fermentation, the same effect was obtained.

Bran and bran were by-products in their respective cereals, and their utilization was limited, and it was unexpected that they could be used as active oxygen scavengers. Furthermore, the possibility of producing an active oxygen scavenger by such a simple treatment is expected to open a new aspect to the use of bran and bran.

The effect of eliminating the active oxygen of the product of the present invention will be described below. First, the effect of the product of the present invention as a superoxide scavenger was examined. The test method was the NBT method.

Preparation of Reagents 0.05M Na ₂ CO ₃ buffer (pH 10.2) 3 mM xanthine solution; 45.64 mg of xanthine is dissolved in a buffer solution to make 100 ml. 3 mM EDTA solution; EDTA.2Na 11
Dissolve 1.7 mg with distilled water to make 100 ml. BSA solution; Bovine Serum Albumin (Sigma) 1
Dissolve 5 mg in distilled water to make 10 ml. 0.75 mM NBT solution; NBT (nitroblue tetrazolium) 61.32 mg was dissolved in distilled water to obtain 10
Make up to 0 ml. Xanthine oxidase solution: Dilute xanthine oxidase with distilled water and adjust it so that the absorbance in the blank test of the operation method (analysis method) described below is in the range of 0.2 to 0.23. 6 mM CuCl ₂ solution; CuCl ₂ · 2H ₂ O
Dissolve 102.29 mg in distilled water to make 100 ml.

Operation method 2.4 ml of Na ₂ CO ₃ buffer solution was placed in a test tube, and xanthine solution, EDTA solution, BSA solution, NBT was added to this.
Add 0.1 ml of each solution. Then, 0.1 ml of the sample solution is added, and after standing at 25 ° C. for 10 minutes, 0.1 ml of the xanthine oxidase solution is added, and the mixture is quickly stirred and incubated at 25 ° C. After 20 minutes, 0.1 ml of CuCl ₂ solution was added to stop the reaction, and the absorbance was measured at 560 nm. For comparison, instead of the sample, superoxide dismutase (Cu, Zn type SOD, activity 3000 to
4000 unit / mg Wako Pure Chemical Industries, Ltd.) The same procedure was performed for 0.1 ml of the aqueous solution, and this value was used as the superoxide elimination rate 1
00. In addition, distilled water is used instead of the sample, and the same procedure is performed to obtain a blank. The measurement results are shown in Table 1.

[0020]

[Table 1] The poured water extract was obtained in Example 1, the organic solvent extract was obtained in Example 3, the alcohol-fermented product after water extraction was obtained in Example 4, and barley bran and wheat bran were: It carried out according to the Example of rice bran.

As described above, it was found that both the water extract and the organic solvent extract have a superoxide scavenging effect.

Next, the thermal stability of the product of the present invention was examined. First, the product of the present invention and SO obtained in Example 1
D was heat-treated at 70 ° C. for 10 minutes, and its superoxide scavenging ability was examined. The superoxide erasing rate was measured by the above method. The results are shown in Table 2.

[0023]

[Table 2] Note: As in Table 1, each extract used was that obtained in each Example.

As described above, it was found that SOD is unstable to heat, whereas all the products of the present invention have excellent thermal stability. From this, it can be said that the active ingredient for eliminating active oxygen in the product of the present invention has excellent stability against heat.

Since the product of the present invention shows extremely remarkable elimination of active oxygen and is safe for sanitary products, it can be used for medicines, cosmetics, foods and the like, and its use will be described. In addition, the following data was conducted for rice bran.

As a medicine, it can be used as an anti-ulcer agent. The test methods for examining the anti-ulcer action of the product of the present invention and the results thereof are shown below. The effect of oral administration of the product of the present invention on restraint water immersion stress ulcer was examined. The method was performed according to the method of Watanabe et al.
That is, 8-week-old male ddY mice were fasted for 24 hours, orally the product of the present invention obtained in Example 1 was orally administered at 0.3 ml / mouse, and after 30 minutes, put into a stress gauge and put in a water at 15 ° C. The protrusions were dipped and restrained water immersion stress was applied. After 5 hours, the cervical vertebrae were dislocated and sacrificed, and the stomach was excised. After that, 1.5 ml of 1% formalin solution was injected into the stomach, and further immersed in the same solution to lightly fix the stomach tissue, and
It was left as it was for 4 hours. After that, an incision was made along the greater curvature, and the length (mm) of the damage occurring in the glandular stomach was measured, and the total sum per animal was expressed as the ulcer index. As a control, the same amount of physiological saline was orally administered 30 minutes before being put into a stress gauge. Fifteen mice each were used. The results are shown in Table 3.

[0027]

[Table 3]

As shown in Table 3, the average ulcer index in mice administered with physiological saline as a control was 60.
On the other hand, the average ulcer index in the mice to which the present invention product was administered was 31.0, clearly showing that the present invention product is effective as an anti-ulcer agent against restraint water immersion stress ulcer by oral administration. It turned out to be. As a result, it was revealed that the product of the present invention directly acts from the gastrointestinal mucosa and exhibits an effective action as an anti-ulcer agent.

The product of the present invention can also be used as a skin treatment agent. The product of the present invention is applied daily to panelists of various skin diseases.
The results are shown in Table 4, which was applied twice a day in the morning and evening to the affected area and continuously applied for 1 month.

[0030]

[Table 4]

As shown in Table 4 above, since this product has various skin-treating effects, it can be seen that it has a fibroblast-activating action and further an antibacterial action.
Moreover, since it is also useful for xeroderma, acne and the like, it can be seen that it also has a moisturizing effect and an effect of appropriately suppressing the increase of sebum, but actually suppresses this moisturizing effect and the increase of sebum. The test methods for examining the action and the results are shown below.

First, in order to demonstrate the strength of the moisturizing effect of the product of the present invention, a single coating test was conducted using a moisture meter (SKICON200). An environment in which the room temperature was 20 ° C. and the relative humidity was 65% was set as a measurement condition, and the panelists were allowed to rest in the above environment for about 10 minutes before the measurement. The test site (both sides) was selected on the flexion side of the forearm where no rash was observed. The panelists were five people who were suffering from xeroderma. The average value of changes in the water content of the stratum corneum between the main test (using the product of the present invention obtained in Example 1) read by a moisture meter and the control test (using water) is shown in FIG. The measurement method of the one-time application test is as follows.

Measurement method 1) A test site of 5 × 5 cm and a control site are set on the forearm flexion side of the panel. 2) Measure the water content of the stratum corneum at each site. 3) Measure the water content of the stratum corneum immediately after coating the sample and after 30, 60, 90 and 120 minutes.

From FIG. 1, it was confirmed that the water content of the stratum corneum of the product of the present invention increased about 10 times as much as that of the control, immediately after coating. In addition, regarding the period from 30 minutes to 120 minutes after the application, in the application site of the present invention, from the control 2 to 120 minutes.
It can be seen that the water content is tripled.

Next, in order to numerically demonstrate the therapeutic effect of the product of the present invention on xeroderma, a water load test was carried out using the moisture meter (SKICON200) before use of the product of the present invention and after use for 2 weeks. .. The panelists were the 5 persons used in FIG. 1, and the measurement conditions were the same as the one-time application test. A control (measurement at a site not coated with the product of the present invention) was always set so that the effect of the seasonal change in the water content of the living horny layer would not influence the effect determination. The stratum corneum water content was shown as the average value of 5 panelists. The result is shown in FIG. As the product of the present invention, the product obtained in Example 1 was used. The measuring method of the water load test is as follows.

Measuring Method 1) The water content of the stratum corneum of the test site is measured. 2) Place one drop of distilled water on the test site, and after 10 seconds, completely wipe off the water drop with dry gauze. 3) Immediately after wiping, the water content of the stratum corneum is measured after 30, 60, 90 and 120 seconds.

As shown in the graph of FIG. 2, by applying the product of the present invention, the water absorption capacity of the skin (the water content of the stratum corneum after 0 seconds of water loading minus the value of the moisture content of the stratum corneum before loading) ),
It can be seen that both the water retention capacity (the curve drawn by the water content of the stratum corneum from 0 to 120 seconds after water loading) is improved at the same time.

That is, the skin before use of the product of the present invention has a very low water content in the stratum corneum before water loading (average 8.0), and the water absorption capacity (average 50.0) is also considerably reduced. In addition, the stratum corneum of the skin of a normal person also has a water-retaining ability, and after 30 seconds of water loading, the stratum corneum of the skin of the normal person returns to the value before the water loading. These results show that the water absorption capacity, the water retention capacity, and the barrier function are all reduced in the measured morbid stratum corneum. On the other hand, the skin after use of the product of the present invention has a two- to three-fold increase in the water content of the stratum corneum and the water absorption capacity before water loading, and the water retention capacity is considerably improved to the same level as that of a normal person. I understood.

From the above, it can be said that the product of the present invention has an excellent action for improving the water content of the pathological stratum corneum and improving the barrier function. Further, when the product of the present invention is evaluated in combination with the moisturizing action obtained from the one-time application test, the product of the present invention increases the water absorption capacity and water retention capacity of the stratum corneum and absorbs a large amount of water from the outside world.
Furthermore, it can be said that it has a moisturizing action of giving the stratum corneum the property of not releasing the once absorbed water.

Further, in order to experimentally demonstrate the secretion inhibiting effect of the sebum amount of the product of the present invention, changes in the sebum amount after washing the face were measured. As panelists, five persons randomly selected from the ones used in Table 4 were used, and the average value of the change in the amount of sebum between the main test (after washing the face and applying the product of the present invention) and the control test (only the face washing) was shown in FIG. It was shown to. As the product of the present invention, the product obtained in Example 1 was used.

As shown in the graph of FIG. 3, it was found that the application of the product of the present invention significantly suppressed the increase in the amount of sebum. The inhibitory effect on the amount of sebum secretion of the product of the present invention also confirmed the preventive and therapeutic effect on acne. The following tests revealed that when the product of the present invention is applied as cosmetics to the skin, it has the effects of smooth skin, fine texture, wrinkle and rejuvenation, and anti-aging effect.

The product of the present invention was applied to the right arm part of the panelist twice a day for one month, and the site of application of the product of the present invention was measured by a dynamic friction meter. The same part of the left arm was used as a control. The panelists were 6 people. The measurement conditions are as follows. Temperature 25 ℃ Humidity 60% Sensor used KES-SE Friction tester SE-2 type (0.5mm piano wire used) Friction static load 50gf Measuring speed 1mm / sec Measuring distance 30mm (Integral effective range 20mm)

The MMD (coefficient of variation) was 0.0189 in the part of the left arm to which the product of the present invention was not applied, but in the part of the right arm to which the product of the present invention was applied for one month, the MMD was
(Coefficient of variation) fell to 0.0060. The average values of 6 persons were almost the same. It is considered that this is because the fluctuation due to the unevenness of the surface was reduced, and from this, it became clear that the texture of the skin became finer and that the wrinkled skin became younger. When MIU (friction coefficient) was also examined at the same time, it was 0.137 before application, but the skin after application for 1 month dropped to 0.081, and the effect of making skin smooth, the effect of softening skin, Furthermore, it has been found that it also has an anti-aging effect.

Further, the absorption spectrum of the product of the present invention was measured in order to demonstrate the ultraviolet absorption effect of the product of the present invention as data. The results are shown in Fig. 4. From FIG. 4, it can be seen that the product of the present invention has a UVB region of 280 to 320 nm and a wavelength of 320 to 320 nm.
Low wavelength region of UVA of 380 nm (about 320 to 3
It was found that the absorption effect was excellent at 40 nm).

Since the product of the present invention has sufficient absorption only in the UVB region, it may be considered that the effect is not sufficient, but the shortest wavelength of ultraviolet rays reaching the surface of the earth is 295.
In terms of nm, the longest wavelength is said to be 320 to 330 nm, and therefore, it can be said that the ultraviolet absorbing ability in a substantial sense is sufficient. In addition, this is about 2. of the natural herb extracts and natural herb components (oil-soluble licorice extract P-U, lycochalcone A, baicalein, etc.) currently used, which have the ability to absorb ultraviolet rays.
It is worth five times the absorption capacity. Also from this, it was proved that the product of the present invention has an excellent ultraviolet absorbing ability.

Further, in order to exemplify the whitening effect of the present invention, a tyrosinase activity inhibitory effect was tested. As an operation method, 1 ml each of a substrate solution (0.04% tyrosine solution) and a buffer solution (McllvaineBuffer pH 6.8) was accurately placed in an absorption cell, and 1 ml each of water and the product of the present invention obtained in Example 1 were obtained. Put each one accurately, stir and mix, and 35 ℃
After 5 minutes, adjust the absorbance scale to the wavelength of 475 nm and perform zero correction, and then add exactly 0.02 ml of tyrosinase solution (5.3 mg of tyrosinase dissolved in 0.9% NaCl solution) and immediately Incubated with agitation. The absorbance at this time was measured over time (every 3 minutes) and is shown in Table 5.

[0047]

[Table 5]

From the measurement results shown in Table 5, it is understood that the product of the present invention has a tyrosinase activity inhibitory action. From this, it can be said that the product of the present invention has a whitening effect. Further, as described above, the product of the present invention also has a moisturizing action enough to be used as a medicine. Therefore, all of the basic effects of cosmetics are satisfied,
It has a wide range of applications such as creams, emulsions, lotions, cleansing, packs and soaps. Also, by drinking the product of the present invention, the same effects as described above can be obtained.

The product of the present invention can also be used as a food preservative, antioxidant, and freshness-retaining agent. As a representative of Gram-positive bacteria, Bacillus subtili causes spoilage of rice and bread.
s, Bacillus cereus, and Escherichia coli as a general indicator of contamination as a representative of Gram-negative bacteria
The antibacterial activity test of the product of the present invention against ia coli and the results thereof are as follows.

As the medium, 10 ml of ordinary agar medium added with 1 ml of the product of the present invention (obtained in Example 5) was used. As a control, 1 ml of water was added instead of the product of the present invention. Incubate at 35 ℃,
The growth state of each bacterium after 24, 48 and 72 hours was observed and shown in Tables 6 to 8.

[0051]

[Table 6]

[0052]

[Table 7]

[0053]

[Table 8] Note 1 Evaluation: −: No growth +: Little growth ++: Development +++: Large growth

As can be seen from Tables 6 to 8, in the medium to which water was added as a control, the growth of all the bacteria was already large at the time of 10-hour culture, whereas the product of the present invention was used. In the medium supplemented with, Bacillus subt
The growth of ilis and Bacillus cereus was completely inhibited even after 72 hours of culture. Also, Es
Growth was observed in cherichia coli by culturing for 48 hours or more, and in this case as well, a large antibacterial effect was recognized as compared with the control.

[0055]

As is apparent from the above data, by extracting rice bran and bran from a water extract or an organic solvent, it is completely safe and stable to heat, has an excellent effect of eliminating active oxygen, and has various effects. It was found that an active oxygen scavenger having an effect can be obtained.

Since rice bran has long been used as livestock feed, vegetable fertilizer,
It is used as a material for pickles or a raw material for rice oil, and its safety is proven. Therefore, the present invention can be used as a pharmaceutical for preventing or treating the above-mentioned diseases, and can also be added to foods, cosmetics and the like to be useful for promoting health and beauty. That is, the present invention has found that the active oxygen scavenger that can be used in a wide range of fields has been proven to be safe, and that it can be easily obtained from bran or bran, which is an inexpensive cereal by-product. It is extremely significant to find a new use application of bran and bran, which has been used only as a by-product with low utility value until now.

[0057]

【Example】

Example 1 To 5 kg of rice bran, 15 liters of warm water at 60 ° C. and 50 g of liquefying enzyme
Was added and stirred well. Then, the temperature was gradually raised, and the mixture was extracted by boiling for 5 minutes and then cooled to 25 ° C. Then, it was squeezed with a squeezing machine to obtain 13 liters of the squeezed liquid (the product of the present invention) and a residual amount of 6.5 kg.

Example 2 To 1 kg of rice bran, 5 liters of 0.1% hydrochloric acid water was added, well stirred, and left for 7 hours. Then, it was squeezed with a squeezing machine and separated into 4.5 liters of compressed liquid and 1.3 kg of residue. This compressed liquid was neutralized with an alkaline liquid to obtain the product of the present invention.

Example 3 To 1 kg of rice bran was added 3 liters of 95% alcohol, and the mixture was stirred well and left to stand. Four days later, it was squeezed with a squeezing machine to obtain 2.5 liters of the squeezing liquid and 1.2 kg of the residue. A portion of 1 liter of this squeezed liquid was added with 500 ml of water, and alcohol was completely removed by a rotary evaporator.
I got ml.

Example 4 To 1 liter of the water extract obtained in Example 1 was added 20 ml of cultured yeast (association No. 9), the temperature was kept at 20 ° C., and alcohol fermentation was carried out for 5 days. Then, filtration was performed to obtain 950 ml of the product of the present invention.

[Brief description of drawings]

FIG. 1 shows a moisture meter (S
It is a graph which shows the result of having performed the coating test once using KICON200).

FIG. 2 is a graph showing the results of a water load test before using the product of the present invention and after using it for 2 weeks.

FIG. 3 is a graph showing the results of testing the change in the amount of sebum when the product of the present invention is applied after washing the face and only when the face is washed.

FIG. 4 shows an ultraviolet absorption spectrum of the product of the present invention.

Claims (1)

[Claims] 1. An active oxygen scavenger characterized by comprising a water extract or an organic solvent extract from rice bran or bran as it is or containing the same.