NL193398C - Antioxidant oil composition and method for its preparation. - Google Patents

Description

1 193398

Antioxidant oil composition and method for its preparation

The present invention relates to oil composition obtained by heating a raw grain material comprising rice germs and / or wheat germ and soybeans at a temperature not higher than 100 ° C, adding koji (aspergillus orizae) and fermenting the heat being material, processing the fermented material into a powder and adding the powdered material to an oil mixture comprising a pasta oil obtained from sesame heated at a temperature not higher than 100 ° C.

Such a composition is known from GB 2,195,889 A. This reference describes an anti-oxidizing composition obtained by fermenting plant seeds, grains and / or germs thereof with microorganisms and adding an oil obtained by heating of plants.

Antioxidants that have an excellent effect when used in the prevention and treatment of various inflammations and chronic diseases such as chronic arthro-rheumatism, thromboplebitis, progressive systemic sclerema, Buerger's disease, Raynaud's disease, persistent skin ulcers and such, which are caused by the destruction of the cellular organization by reactive oxygen and lipid peroxides produced in the human body, have recently become of interest.

Research has clarified that low molecular weight antioxidants such as flavonoids, polyphenols, tannins, tocopherols, vitamin B1 and the like are present in plants, and that by administering them orally as a drug or health food, the production of the above reactive oxygen suppressed in lipid peroxide in the body (see Journal of Japan Pharmacist Association,

No. 39, Vol. 12, Supplement, "Bioavailability of SOD (Superoxide Dismutase) and crude drug", biz. 1097-1199, published December 1, 1987).

Japanese Patent 1,366,268 discloses a vegetable foodstuff obtained by mixing green tea powder with fried brown rice powder and soybean powder, adding a small amount of koji fungus to the mixture, which mixture is then placed in a mixture for about 4 days. mixture of sesame oil and soybean oil is kept to extract the effective ingredients, then the precipitates are removed by centrifugation, and the remaining oily compound is enclosed in a gelatin capsule or the like.

However, the vegetable foodstuff of Japanese Patent 1,366,268 has a low anti-oxidant activity, which is caused by the way in which the raw vegetable material is processed.

GB 2,195,889 A describes an antioxidant composition with increased activity obtained by gently heating plant seeds, grains or germs thereof so that they do not burn, adding a microorganism to the heated plant material to ferment it, and adding the fermented material to a vegetable oil obtained from a plant which was also gently heated. Optionally, vitamin C, vitamin C derivatives or a plant containing them are added to the composition. As the plant seed, a seed containing antioxidants such as the aforementioned flavonoids, polyphenols, tannins, tocopherols, vitamin B1 and the like is used, including rice, wheat, barley, soybeans, azuki beans (Phascolus radiatus L, var. Typicus Prain), corn, pearl barley (hatomugi) (Coix 40 Lacryma-jobi var.muyen Stapf L), peas. As the vegetable oil, sesame oil, soybean oil, cottonseed oil, corn oil, safflower oil, primrose oil, rice bran oil, rapeseed oil, olive oil and the like are used.

However, it has been found that although the above antioxidant composition markedly suppresses the production of reactive oxygen and lipid peroxide in vitro, these antioxidant activities do not occur sufficiently in the human body.

45 It has now been found that by incorporating in an antioxidant composition according to GB 2,195,899 A in addition to the oil obtained from heated plant material, an oil obtained from crude sesame, and adding a certain ratio between oil mixture and oil composition. To fit, an oil composition is obtained which has a surprisingly high anti-oxidant activity and a high cell penetration capacity.

The invention thus provides an oil composition as described in the preamble, characterized in that the oil mixture further comprises an oil obtained from crude sesame, and the ratio of the oil mixture to the total amount of the powdered material and the oil mixture is 60 to 95% by weight.

As raw material containing antioxidants in large amounts, in addition to rice sprouts, wheat germ and 55 soybeans, the plant species described in GB 2,195,899 A can be mentioned, i.e. barley, azuki beans, corn, pearl barley (hatomugi), peas and the like. In addition to rice sprouts, wheat germ and soybeans, rice bran, pearl barley (hatomugi) and wheat are particularly suitable. Therefore, the raw grain material 193398 2 may contain at least one material selected from rice bran, pearl barley (hatomugi) and wheat in addition to the rice germ and / or wheat germ. The amount of the germs (rice germ and / or wheat germ) in the raw grain material should be at least more than or equal to 1% by weight.

The antioxidants present in the raw grain material described above together with 5 other compounds form a complicated macromolecular polymer structure. In this macromolecule, the antioxidants have no activity, so it is necessary to break the bonds holding the macromolecule together by gentle heating or the like, thereby releasing low molecular weight antioxidants. However, if the heating temperature is too high, the low molecular weight antioxidants are deactivated, so that it is necessary to take this fact into account when choosing the heating conditions.

To take this into account it is necessary that the raw wheat material is heated at a temperature not higher than 100 ° C, heating slowly and for a sufficient time. In detail, the raw grain material is placed in a vessel made of ceramic material such as pottery or the like which radiates with infrared rays from the far-red region, which preferably have a wavelength of 5 to 14 µm, while heating is performed with slow stirring keeping the temperature at about 90 to 96 ° C.

The heating time varies depending on the types of grains, so it cannot be readily set, but it is preferably from about 30 minutes to 3 hours. Incidentally, the heating method is not limited to the above method, provided that the antioxidants in the raw grain material are sufficiently released in the form of low molecular compounds, while the deactivation thereof is prevented.

After the heat treatment described above, koji (Aspergillus orizae) is added to the crude material to ferment it. The fermentation is preferably carried out at 20 to 36 ° for about 2 to 6 days. When the fermentation is carried out in a fermenter, about 2 to 3 hours is sufficient. Fermentation is carried out to promote the release and conversion to low molecular weight compounds of the antioxidants in the raw grain material, and through this fermentation step, the activity of the antioxidant composition can be significantly increased compared to raw material that is only subject to heat treatment.

Then the fermented material is ground to convert it into a fine powder. Grinding can be performed with a commercially available grinder. However, some types of grinders generate high temperatures during use, which deactivates the antioxidants. It is therefore preferred to use those grinders that do not generate a high temperature in use. For example, a stone mill can be used.

Then, the oil is prepared by mixing an oil obtained from hot steam (hereinafter referred to as sesame paste oil hereinafter) with an oil obtained from crude sesame, in an appropriate ratio, and the above fine powder is added to the oil mixture . The sesame paste oil is an oil obtained by heating crude sesame at a temperature not above 100 ° C, slowly and for sufficient time, and grinding and pressing it, but the fine solids formed by grinding the sesame will remain as is, so the product will look like a 40 paste. This sesame paste oil contains the low molecular weight antioxidants in high concentration, and by using it it is possible to obtain the oil composition with high activity of the antioxidants.

However, this sesame paste oil has a high viscosity, and the size of an oil droplet is also large, so that the composition in which only the sesame paste oil is mixed with the fine powder described above has poor penetration into the insides of cells in a diseased portion. When 45 sesame oil obtained from crude sesame is added to the sesame paste oil, the droplet size may be small, and the penetration into the insides of the cells in a diseased portion is improved.

The oil recovered from crude sesame is an oil obtained by grinding and squeezing crude sesame and removing the solids, and it is commercially available as a regular sesame oil. Since the mixing ratio of the sesame paste oil to the regular sesame oil also differs by 50 depending on the amount of fine powder to be added, it cannot be defined in all cases. The mixing ratio is preferably 1 to 3 wt. parts of the regular sesame oil per wt. part sesame paste oil. The ratio of the oil mixture to the fine powder added thereto is such that the mixed oil makes up 60 to 95% by weight of the total of both. If the oil mixture is less than 60% by weight, the ability to pass through the cell membrane is small. On the other hand, if the oil mixture is more than 95% by weight, the concentration of antioxidants as effective compounds is low, so that the effect of suppressing the production of reactive oxygen and a lipid peroxide also decreases.

3 193398

The oil composition of the present invention, which is administered orally, can be prepared by adding the fine powder to the oil mixture by enclosing the whole once the oil mixture has been prepared in a gelatin capsule or the like. Preferably, however, the material mixture can be allowed to mature at 20 to 35 ° C, more preferably at 28 to 30 ° C for about 1 week, for about 3 to 30 days.

By maturing the mixture, the release and conversion into low molecular weight compounds of the antioxidants is further promoted, as the koji remains in the fine powder, and the antioxidants adhere well to the oil mixture, so that the activity and osmotic ability to the inside of the cells of the reactive compounds is further improved.

The oil composition of the present invention may also include a raw grain material, consisting of rice sprouts or wheat sprouts and soybeans, and optionally at least one product selected from rice bran, pearl barley or wheat, which as such is ground into a fine powder without heating and to ferment, and is added to the oil mixture together with the above-described fine powder which has been heated and fermented.

In the fine powder of the raw grain material which has not been heated and fermented, the antioxidants as effective compounds have not been released and have been converted into low molecular weight compounds. However, when added to the oil mixture along with fine powder that has been heated and fermented, the release and conversion to low molecular weight compounds of the antioxidants continues even after encapsulation, due to the fact that the koji in the fermented fine powder remains, so that there is the advantage that the effect of suppressing the production of reactive oxygen or lipid peroxide is maintained for a long period of time.

In contrast, in the oil composition using only heated and fermented fine powder, due to enzyme reactions and the like, which continue continuously even after encapsulation, the antioxidant compositions as effective ingredients are gradually decomposed so that the effective period is short compared to the composition in which the untreated fine powder is added.

However, if the amount of untreated fine powder is too large, the activity of the effective ingredient decreases. Therefore, it is preferred that the untreated fine powder be about 0.5 to 1 wt. part per wt. part of the heated and fermented fine powder. Incidentally, when two types of fine powder are included, the amount of the oil mixture is 60 to 95% by weight of the total amount of powder and oil mixture.

In the oil composition of the present invention thus obtained, both the activity of the antioxidants and the osmotic ability to enter the cells are high, and, as is also apparent from the results of the clinical tests to be described below, it shows surprising effects against various inflammations and chronic diseases, whereby conventional anti-inflammatory agents have been unable to obtain sufficient therapeutic effect.

The oil composition of the present invention can be incorporated into gelatin capsules or the like, and administered orally as a medicament. In addition, in case of abnormal pigmentation such as dermatitis, chloasma, freckles and the like or wrinkles and the like, the oil can be applied directly to an affected area. In addition to dermatitis and the like, the oil composition can be used in aging and chronic diseases such as chronic arthroreuma, thromoplebitis, progressive systemic sclerema, Buerger's disease, Raynaud's disease, persistent skin ulcers and the like. The composition of the invention can also be used in the treatment and prementia of various other pollution-caused diseases, burns, superficial wounds, fatigue, adverse effects of excessive alcohol consumption, constipation and the like.

In addition, the use of the oil composition of the present invention is not associated with side effects, since only grain, koji and sesame are used as the starting material, so that it is possible to use the composition as a health food for maintaining and increasing health. It is not necessary to say that in the preparation of a composition it is possible to add other drugs or ingredients useful in health, such as various vitamins, minerals and the like, as well as fragrances, flavors, dyes and the like.

55 Example

Rice germ, soybeans, rice bran, hatomugi {pearl barley) and wheat each of one part was put in a vessel made of pottery and emitted infrared rays from the distant region with a 193398 4 wavelength from 4 to 14 µm, and was stirred for 3 hours stirring gently to avoid scalding heated at 90 to 96 ° C. Koji was then added in an amount of 3% of the total amount, and fermented at 36 to 40 ° C for 72 hours, after which the fermented material was processed into a fine powder using a stone mill. In addition, the above-mentioned five kinds of grain materials in a raw form, in a non-heated and fermented state were milled with a stone mill to obtain the same amount of fine powder. Furthermore, sesame was heated in the same manner as the above materials, and after being ground with a stone mill, squeezed to obtain a sesame paste oil.

Then, 28.1 wt. parts of sesame paste oil mixed with 48.9 wt. parts of a commercially available sesame oil, to which mixture 11.5 wt. parts of the heated and fermented fine powder and 11.5 wt. parts of the untreated fine powder were added, and the resulting mixture was stirred homogeneously. The mixture was allowed to mature at 28 ° C for one week to obtain an oil composition, which was then placed in gelatin capsules.

Effect of the Invention [I] In Vitro Assay 1. For an in vitro assay of the ability to reach and penetrate oily spots, the oil composition of the invention was added to a TBA (thiobarbituric acid) reaction system, in which an oily unsaturated fatty acid, decosahexacetic acid (C21H31COOM), reacted with reactive oxygen generated by ultraviolet radiation to produce lipid peroxide, and the degree of suppression of lipid peroxide production was determined.

To 0.1 ml of decosahexaenoic acid diluted 100 times with ethanol, 1.8 mg / ml of the test sample of the oil composition of the present invention was added, and the amount of lipid peroxide produced was determined by the TBA reaction. In the TBA reaction, 0.2 ml of 7% sodium dodecyl sulfate, 2 ml of 0.1 N HCl, and 0.3 ml of phosphorus tungsten acid (H3 [P (W, 010) 4] .14H2 O) were mixed, t.5, after which 1, 8 mg / ml of the test sample was added to the mixture, then 1 ml of a reagent in which 0.67% TBA was mixed with acetic acid in a ratio of 1: 1 was added, and the assay was performed with excitation at 515 nm and emission at 553 nm using a fluorescence spectrophotometer.

As comparative examples, assays were similarly made with the vegetable nutrient of Japanese Patent 1,366,268 (Comparative Example 1) and the antioxidant composition described in Japanese Unexamined Patent Application Laid-Open No. 63-79834 (Comparative Example 2) . Results are shown in Table 1.

TABLE 1 35 -; -

Test sample Mean (value after 6 minutes)

Control (UV +) 461 ± 62 40 Oil composition according to the invention (1.8 mg / ml) 121 ± 13 ***

Comparative Example 1 (1.8 mg / ml) 271 ± 34 **

Comparative Example 2 (1.8 mg / ml) 358 ± 45 * 45 UV +: Ultraviolet Radiation *: 0.01 <P <0.05 (vs control) **: P <0.01 (vs control) *** : P <0.0001 (vs control)

Test Results 50 Although each of the samples tested suppressed the generation of the lipid peroxide (TBA reacting compounds) from the unsaturated fatty acid (decosahexanoic acid) by irradiation with ultraviolet radiation (102), the oil composition of the present invention significantly suppressed this generation (P <0 , 0001). This result confirms the fact that the oil composition of the present invention has higher anti-oxidant activity and penetration to the interior of the cell in which a disease occurs than the conventional compositions (Comparative Examples 1 and 2). ).

2. A [3,4-3H2] antioxidant composition was prepared in which the low molecular weight antioxidants present in the oil composition of the present invention were labeled with the isotope (3H), in a test tube was placed containing human tissue, and the amount (cpm) of [3H2] bound to its cell membrane was determined with a scintillation counter, examining the ability to reach the cell membrane (labeled tritiated thymidine was 2 Ci / mM).

The same experiments were also performed for comparative examples 1 and 2. The results are shown in table 2. As can be seen from table 2, the oil composition of the present invention had the highest affinity for the cell membrane compared to comparative examples 1 and 2.

TABLE 2 10 -

Test sample cpm incorporation

Control 15643 cpm / 103 cells

Oil composition of the present invention (1.8 mg / ml) 56372 cpm / 103 cells Comparative Example 1 (1.8 mg / ml) 33451 cpm / 103 cells

Comparative Example 2 (1.8 mg / ml) 30567 cpm / 103 cells 3. The oil composition of the present invention was subjected to an ultrasonic treatment and added to a reactive oxygen generating system (neutrophil and xantine-xantine oxidase) so that the a living body converted to a concentration of 1.6 mg / ml, and the three types of reactive oxygen (O 2 ', H 2 O 2 and OH *) were determined as compared to the situation when nothing was added (control). The concentration converted to a living body is an amount believed to be present in the blood when a usual daily dose (9 g in the case of the oil composition of the present invention) is taken up in a living body.

The methods of determination for the three types of reactive oxygen are as follows.

For the determination of 02 ', the amount of reduction of ferric cytochrome C by 02' was determined at a wavelength of 550 nm with a Beckman spectrophotometer, and converted to the amount of 02

For the determination of H202, use was made of the fact that H202 reduces the fluorescence generated by scopoletin in the presence of peroxidase, using scopoletin and peroxidase, determining the decrease in fluorescence and scopoletin with a fluorescence spectrophotometer of Hitachi Ltd. with an excitation at 370 nm and an emission at 460 nm.

For the determination of OH *, use was made of the fact that α-keto-methiol-butanoic acid (KMB) reacts with OH- to form ethylene gas, the method involving quantifying the amount of ethylene gas with a gas chromatogram made by a gas chromatograph Hitachi Ltd. and the amount of ethylene was calculated back to the amount of OH *. The same tests were also performed for comparative examples 1 and 2. The results are shown in Table 3.

TABLE 3 40 -

Reactive Oxygen

Test sample 02 'H202 OH *

Control 1.532 nmol 485 pmol 854 pmol 45 Oil composition of the invention (1.8 0.589 nmol 161 pmol 283 pmol mg / ml)

Comparative Example 1 (1.8 mg / ml) 1.285 nmol 403 pmol 707 pmol

Comparative Example 2 (1.8 mg / ml) 0.231 nmol 538 pmol 108 pmol 50

Test results:

The antioxidant activity of the oil composition of the present invention was slightly less than that of Comparative Example 2, but higher than that of Comparative Example 1. As can be seen from Tables 1 and 2, Comparative Example 2 contains a smaller amount of oily compounds, so that 55 it has a poor ability to reach the cell in which the disease takes place.

Hence, viewed as a whole, the oil composition of the present invention, the antioxidant activity of which is high and the ability to penetrate inside the cell of a

Claims (3)

193398 6 most affected, most suitable as an antioxidant. [II] Clinical Trial The therapeutic effect was studied in 96 patients with autoimmune diseases, collagen diseases such as chronic arthroreuma, haemastenosis, nephrititis, hepatocirrhosis, chloasma, freckles and the like, who have not yet responded or were exacerbated with the use of non-steroidal antiphlogistic drugs , steroids and the antioxidant composition of Comparative Example 2. The results are shown in Table 4. TABLE 4 10 - Test Sample Disease Oil Comparative Comparative Theorem of Example 1 Example 2 Invention (1.8mg / ml) (1, 8mg / ml) (1.8mg / ml) Chronic arthroreuma 13/18 (72%) 0/7 (0%) 4/14 (28%) Angiitis 3/5 (60%) 0/3 (0%) 1/4 (25%) 20 Progressive systemic sclerema 9/12 (75%) 2/10 (20%) 6/12 (50%) Dermatomyosytis 6/8 (75%) 2/12 (16%) 6/13 (46%) Thromboplebitis 2 / 3 (66%) 0/4 (0%) 1/5 (20%) Buerger's disease 6/8 (75%) 0/3 (0%) 3/6 (50%) Raynaud's disease 10/12 ( 83%) 3/8 (37%) 7/12 (58%) 25 Crus varicosis 8/11 (72%) 4/9 (44%) 11/15 (73%) Stubborn skin ulcers 3/5 (60%) 1/5 (20%) 2/6 (33%) Chloasma, freckles 11/14 (78%) 4/12 (33%) 17/25 (68%) The percentages refer to the percentage of patients who recover. Oil composition obtained by heating a raw grain material comprising rice germs and / or wheat germ and soybeans at a temperature not higher than 100 ° C, adding koji (Aspergillus Orizae) and fermenting the material that has been heated, processing the fermented material into a powder, and adding the powdered material to an oil mixture comprising a paste oil obtained from sesame heated at a temperature not higher than 100 ° C, characterized in that the oil mixture further comprises an oil obtained from crude sesame, and the ratio of the oil mixture to the total amount of the powdered material and the oil mixture is 60 to 95% by weight.