KR20160139170A - Manufacturing Method of Extracts Powder of Black Rice Bran And Extracts Powder of Black Rice Bran By The Same - Google Patents
Manufacturing Method of Extracts Powder of Black Rice Bran And Extracts Powder of Black Rice Bran By The Same Download PDFInfo
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- KR20160139170A KR20160139170A KR1020150073576A KR20150073576A KR20160139170A KR 20160139170 A KR20160139170 A KR 20160139170A KR 1020150073576 A KR1020150073576 A KR 1020150073576A KR 20150073576 A KR20150073576 A KR 20150073576A KR 20160139170 A KR20160139170 A KR 20160139170A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
- A23L7/198—Dry unshaped finely divided cereal products, not provided for in groups A23L7/117 - A23L7/196 and A23L29/00, e.g. meal, flour, powder, dried cereal creams or extracts
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
- A23L7/115—Cereal fibre products, e.g. bran, husk
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
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- A23V2250/21—Plant extracts
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract
The present invention relates to a method for preparing an extract of black rice microgranular extract for prevention or treatment of antiinflammation and immunological diseases containing an extract of black rice microgranules as an active ingredient and to a method for extracting black rice microgranules according to the method. More specifically, And extracting powder by centrifugation, filtration, concentration, sterilization, cooling, and spray drying. The present invention also relates to a process for preparing the extracted rice bran rice, and a method for preparing the extracted rice bran rice.
Description
The present invention relates to a method for preparing an extract of black rice microgranular extract for prevention or treatment of antiinflammation and immunological diseases containing an extract of black rice microgranules as an active ingredient and to a method for extracting black rice microgranules according to the method. More specifically, And extracting powder by centrifugation, filtration, concentration, sterilization, cooling, and spray drying. The present invention also relates to a process for preparing the extracted rice bran rice, and a method for preparing the extracted rice bran rice.
Black rice, which is a kind of colored rice, is processed into various types of food due to its unique color and flavor, and its consumption is gradually increasing. The aroma of black rice is derived from alcohol components such as ethanediol and guaiacol and ketones such as hexadecanoic acid, hexanal and acetic acid, aldehydes and organic acids. And pigment components are reported to be the main components of glycosides such as cyanidin-3-glucoside (C3G) and peonidin-3-glucoside (P3G).
In particular, the pigment component of black rice contains polyphenol compounds having various structures and molecular weights, and such polyphenol compounds have been confirmed to have physiological activities such as antioxidant, antimicrobial and anticancer properties. In addition, black rice is supplied in brown rice, has higher dietary fiber content than ordinary brown rice, has unique flavor, protein, vitamin B, and mineral content.
Recently, as rice processed food is modernized and diversified, interest in research of rice as a food processing raw material is increasing with the development of a new processed food which mainly uses rice. However, until now, the use of black rice has remained at a level of simple processing, and an anthocyanin pigment, which is reported to be a substance promoting anti-inflammation and immunological activity, has not been developed. Therefore, there is a demand for a highly functional product that enhances anti-inflammatory and immunological activity by using an extract derived from black rice.
On the other hand, as a method of extracting an active ingredient containing an anthocyanin pigment reported as a substance promoting antiinflammatory and immunological activity of rice black rice, the Korean rice patent publication No. 2012-0098070 discloses a method of extracting black rice gyoza using an index Discloses an antioxidant functional composition comprising an extract of the ash extract, which is prepared therefrom, as an active ingredient, and Korean Patent No. 1163072 discloses a method of purifying and pulverizing the extract of Aronia fruit. However, the above methods are produced in a small-scale laboratory unit, and the extraction yield is low. When the above-mentioned index of extraction method is used, there is a disadvantage that expensive facility and maintenance cost can be obtained. In the latter case, It is produced by using sulfuric acid in the acidification process.
Therefore, the inventors of the present invention have found that it is possible to produce a black rice micropowder powder which can maximize the effective ingredients such as anthocyanin pigment which enhances the anti-inflammatory and immunological activity possessed by the black rice micropowder when spray-dried under specific conditions after hot water extraction Thereby completing the present invention.
It is an object of the present invention to provide a method for preparing an extract of black rice rice bran for the prevention or treatment of anti-inflammatory and immunological diseases, which comprises extracts extracted from black rice bran, as an active ingredient, and the resulting black rice bran extract powder.
In order to accomplish the above object, there is provided a method for producing a fermented soybean milk, comprising the steps of adding water to a black rice micropowder powder to extract hot water, centrifuging the extracted hot water extract, concentrating the filtered extract, A step of cooling the sterilized extract, and a step of spray-drying the cooled extract to prepare an extract powder.
In the step of extracting hot water, it is preferable that hot water extraction is performed for 1 to 3 hours by adding water at 50 to 70 ° C to the black rice micropowder powder.
The centrifugation and filtration may be performed by centrifuging with a decanter to remove the extracted foil, and then filtering the centrifuged liquid from which the extracted foil has been removed with a bag filter.
The concentration step can be carried out by natural recycling to 50 to 55 ° C. so that the evaporation amount is 500 kg / hr.
The sterilization step is preferably sterilized in a hemispherical concentrator at 90 DEG C for 10 minutes.
Preferably, the cooling step is performed at a temperature of 70 DEG C or lower.
In addition, the step of spray-drying the cooled extract to produce an extract powder may include the steps of: cooling the extracted extract at an inlet temperature of 180 to 200 ° C, an outlet temperature of 80 to 100 ° C, The nozzle press is preferably spray-dried using an electric heater having a pressure of 40 to 70 bar.
In addition, the present invention provides a method for preparing brown rice extract from rice bran.
There is provided a method for preparing anti-inflammatory and immunological diseases, comprising the steps of:
which is capable of inhibiting iNOS and
The details of other embodiments are included in the detailed description and drawings.
The method of the present invention for preparing an extract of B. thunbergii for the prevention or treatment of inflammation and immune diseases is suitable for mass production by hot water extraction, centrifugation, filtration, concentration, sterilization, cooling and spray drying.
In addition, the method of the present invention can further inhibit iNOS and
FIG. 1 is a flowchart showing a process for producing a black rice bran extract prepared according to an embodiment of the present invention.
2 is a photograph of a black rice bran extract powder according to Example (a) and Comparative Example (b) of the present invention.
FIG. 3 is a graph showing the effect of black rice bran extract on macrophages according to an embodiment of the present invention.
4 is a graph showing the effect of black rice extract of rice bran on iNOS and
FIG. 5 is a graph showing the effect of adding black rice bran extracts according to an embodiment of the present invention to an immune cell proliferation effect. FIG.
FIG. 6 is a graph showing the evaluation of NK cell proliferative activity by a black rice bran extract according to an embodiment of the present invention.
FIG. 7 is a graph showing changes in cytokine production amount in the treatment of black rice rice bran extract according to an embodiment of the present invention.
8 is a graph showing the effect of C3G on macrophages according to an embodiment of the present invention.
9 is a graph showing the influence of C3G on iNOS and
FIG. 10 is a graph showing the effect of C3G on immune cell proliferation (growth promotion effect on B and T cells) according to an embodiment of the present invention.
11 is a graph showing the evaluation of NK cell proliferative ability by C3G according to an embodiment of the present invention.
12 is a graph showing changes in IL-6 production amount with respect to C3G treatment according to an embodiment of the present invention.
13 is a graph showing the effect of red ginseng extract on macrophages according to an embodiment of the present invention.
FIG. 14 is a graph showing the effect of black rice bran extract and red ginseng extract on iNOS and
FIG. 15 is a graph showing the effect of the addition of black rice bran extract and red ginseng extract on the immune cell proliferation effect according to an embodiment of the present invention.
FIG. 16 is a graph showing the evaluation of NK cell proliferative activity by black rice microbrewroth extract and red ginseng extract according to an embodiment of the present invention. FIG.
17 is a graph showing changes in cytokine production amounts of black rice bran extract and red ginseng extract treatment according to an embodiment of the present invention.
BRIEF DESCRIPTION OF THE DRAWINGS The present invention is capable of various modifications and various embodiments, and specific embodiments are illustrated in the drawings and will be described in detail in the detailed description. It is to be understood, however, that the invention is not to be limited to the specific embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.
The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In the present application, the terms "comprises" or "having" and the like are used to specify that there is a feature, a number, a step, an operation, an element, a component or a combination thereof described in the specification, But do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.
The terms first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another.
The present invention relates to a method for preparing an extract of black rice rice bran extract for the prevention or treatment of anti-inflammatory and immunological diseases containing an extract of black rice rice bran as an active ingredient. More specifically, , Followed by concentration, sterilization, cooling, and spray drying to produce an extract powder.
The anti-inflammatory and anti-inflammatory and immunosuppressive anti-microbial powders for use in the present invention include extracts extracted from black rice bran as an active ingredient.
The black rice micropowder is a by-product obtained in the process of rice brown rice finishing to black rice. The black rice micropowder is washed with water to remove foreign matter, and freeze-dried or spray-dried in a freeze dryer to be pulverized.
Hereinafter, the present invention will be described in detail.
FIG. 1 is a flowchart showing a process for producing a black rice bran extract prepared according to an embodiment of the present invention.
In order to accomplish the object of the present invention, the present invention provides a method for producing a fermented beverage comprising the steps of adding water to a black rice goblet powder to effect hot water extraction (S10), centrifuging and filtering the extracted hot water (S20) (S30) of concentrating the filtered extract (S30), sterilizing the concentrated extract (S40), cooling the sterilized extract (S50), spray drying the cooled extract to prepare an extract powder (S60).
In step (S10) of adding water to the black rice micropowder powder, hot water extraction is performed by adding water to the black rice micropowder powder at 50 to 70 ° C for 1 to 3 hours. For example, the black rice micropowder powder and water are mixed at a weight ratio of 1:15 to 25, preferably 1:18 to 20, and subjected to hot water extraction at 50 to 70 ° C for 1 to 3 hours, preferably 1 hour, .
If the temperature is lower than the lower limit, the anti-inflammation and immunological activity can not be expected. If the temperature is above the upper limit, anti-inflammation and immunological activity may be poor. In addition, when the time of extraction of rice bran under the above-mentioned lower limit is low, anti-inflammation and immunological activity can not be expected. If the above-mentioned upper limit is exceeded, anti-inflammation and immune activity can not be expected and toxicity can be caused. Further, for example, water can be added to the black rice micropowder powder, and an extraction tank can be used for hot water extraction, and extraction can be performed by rotating the impeller at 21.93 Hz.
The step (S20) of centrifuging and filtering the extracted hot-water extract may include centrifuging with a decanter to remove the extracted foil, and filtering the centrifuged liquid from which the extracted foil has been removed with a bag filter have. According to one embodiment, since the characteristic of the raw material for black rice microgranules is a fine powder, centrifugation (main motor 52 Hz, vehicle speed motor 38 Hz, feed pump 4 rpm) is performed using a decanter rather than a general filtration process, It is possible to remove the foil and increase the recovery rate of the extract. The extract can be centrifuged and then filtered using a filter to increase clarity (transparency). For example, the filtration filter can be re-filtered using a 55 μm bag filter to increase clarity.
Next, it is preferable that the step (S30) of concentrating the filtered extract is carried out by natural circulation at 50 to 55 DEG C such that the evaporation amount is 500 kg / hr.
The step of sterilizing the concentrated extract (S40) may be sterilized using a concentrator to remove contamination of microorganisms. The sterilization can be carried out in a hemispherical concentrator at a temperature of 80 to 95 ° C for about 5 to 15 minutes. More preferably, it is preferable to sterilize the hemispherical concentrator at 90 DEG C for 10 minutes. When the extraction amount is small, the concentration step may be omitted and the drying step may be carried out.
The step of cooling the sterilized extract (S50) comprises cooling the sterilized extract to 70 DEG C or less.
The thus obtained black rice extract of rice bran can be used as an extract itself or can be used as powder for convenient use. In order to powder the black rice bran extract, the black rice bran extract powder can be prepared by freeze drying, spray drying or hot air drying. The method of pulverizing the black rice extract of rice bran can be carried out by reducing the volume of the extract by concentration under reduced pressure, followed by pulverization using one selected from the group consisting of freeze drying, spray drying and hot air drying. For example, in the case of freeze-drying, the treatment can be carried out at a temperature of -30 ° C to 30 ° C for 2 to 4 hours, but the burden of the process cost is large during lyophilization, and there is a possibility that the pulverization process is added after drying .
According to an embodiment of the present invention, the step (S60) of spraying and drying the cooled extract to prepare an extract powder may include: cooling the extracted extract at an inlet temperature of 180 to 200 ° C, The temperature is preferably 80 to 100 DEG C, and the nozzle pressure is preferably spray-dried using an electric heater having a pressure of 40 to 70 bar. During spray drying, a magnet rod (12,000 Gaus) can be installed at the end of the vibrating body to remove metal impurities. In the case of spray drying, powdering is performed well without addition of excipients.
Another aspect of the present invention is to provide a method for producing rice bran extract powder according to the present invention. It is also characterized in that it is a black rice extract of rice bran for the prevention and treatment of anti-inflammatory and immunological diseases, which is prepared according to the above-mentioned method for producing rice bran extract. The RBC extract powder prepared according to the method of the present invention can inhibit iNOS and
It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the present invention. Such variations and modifications are intended to be within the scope of the appended claims.
EXAMPLES: Preparation of Brown Rice Bran Extract Powder Using Spray Drying
A large amount of raw black rice was extracted, centrifuged and filtered, concentrated, sterilized, cooled, spray dried and pulverized.
First, 90 kg of the black rice micropowder powder prepared from the above-mentioned black rice bran extract was mixed with 1,800 kg of water at a weight ratio of 1:20, and the mixture was stirred at 70 ° C. for 1 hour using an extraction tank (capacity: 2 tons) And extracted by hot water. In order to filter the extracted hot-water extract, centrifugation (main motor 52 Hz, vehicle speed 38 Hz, feed pump 4 rpm) was performed by using a Decanter to remove the extracted foil, thereby increasing the recovery rate of the extract. The centrifuged filtrate was re-filtered through a bag filter (55 μm).
Thereafter, the filtered extract was concentrated to a temperature of 50 to 55 ° C using a natural circulation concentrator so that the evaporation amount was 500 kg / hr. Then, it was sterilized at 90 DEG C for 10 minutes using a hemispherical concentrator, and then cooled to 70 DEG C or less.
The cooled extract is heated at an inlet temperature of 180 to 200 ° C, an outlet temperature of 80 to 100 ° C, and a nozzle press of an electric heater of 40 to 70 bar Followed by spray drying. At the time of the spray drying, a magnet rod was provided at the end of the vibrating body to remove metal foreign substances, and a black rice bran extract powder was obtained.
Comparative Example: Preparation of Black Rice Bran Extract Powder by Freeze-Drying
50 g of rice bran powder and 20 times of water (1 kg) were added and mixed. The resulting extract was extracted with hot water using a constant temperature water bath at 70 캜 for 2 hours, filtered and filtered through a 55 탆 bag filter. Thereafter, lyophilized rice bran extract powder was prepared by freeze-drying at -30 ° C to 30 ° C, 0 Torr pressure, and 120 minutes to 9999 minutes according to the temperature, pressure, and time conditions shown in Table 1 below.
[Table 1] Freeze drying conditions
2 is a photograph of a black rice bran extract powder according to Example (a) and Comparative Example (b) of the present invention. FIG. 2 (a) is a granular black rice micropowder extract powder which is prepared according to one embodiment and has good flowability and solubility. FIG. 2 (b) shows a reddish brown light scattering It is a black rice germ extract powder.
Experimental Example 1: Measurement of anthocyanin content and yield of extract powder prepared according to Examples and Comparative Examples
<1-1> Determination of anthocyanin content and yield of the extract powder prepared according to the examples
The anthocyanin content and yield of the extract powder prepared according to the examples were measured (see Table 2).
[Table 2] Anthocyanin content and yield of extract powder prepared according to the examples
As shown in Table 2, the C3G yield (%) was 13.43% based on the raw material.
<1-2> Anthocyanin content and yield of extract powder prepared according to Comparative Example
The anthocyanin content and yield of the SV01 to SV09 extract powder prepared according to the comparative examples were measured and the average values were shown in Table 3 below.
[Table 3] Anthocyanin content and yield of extract powder prepared according to Comparative Example
As shown in Table 3, the C3G yield (%) was 9.18% based on the raw material.
When the results of Table 2 and Table 3 were compared, it was found that the yield (23.58%) of the extract powder through the spray drying process of the bulk extract of the above Example was higher than that of the brown rice extract powder by freeze- The yield of anthocyanin (C3G) is 13.43%, which is slightly lower than that of anthocyanin (25%) of the extract powder prepared in Comparative Example.
Therefore, the method of mass-production of black rice gruel according to the embodiment of the present invention is superior to the extraction method by a small amount of freeze-drying process according to the comparative example.
Experimental Example 2: Evaluation of immunological activity of the black rice extract (Pilot, Plant scale) prepared according to the examples
The extracts of black rice rice bran extract prepared by the examples were extracted from the pilot scale and the plant scale.
All of the black rice microgranular extracts prepared according to the above-described examples were dried by spray drying (SD) method and dissolved in experimental DMSO. The final concentration of the extracts was ug / ml.
(a) Effect on RAW 264.7 cell viability
The cytotoxicity of the black rice extract prepared according to the above example was evaluated by MTT assay using macrophage RAW 264.7 cells.
The concentration interval was set in comparison with the above experiment. The MTB assay was performed by culturing the cells for 24 hours after adding the black rice bran extract prepared according to the above examples.
As shown in FIG. 3, both of the black rice rice bran extracts prepared according to the above-described examples of two different sizes of extracting scale showed no toxicity at 0.1 to 1 ug concentration and showed the ability to proliferate.
(b) NO and H 2 O 2 Effect on production
The effect of the black rice extract on the production of NO and H 2 O 2 related to immunological activity was evaluated.
As shown in FIG. 4, when the two extracts were treated alone, the expression of iNOS protein was slightly increased by the pilot extract, and the expression of the iNOS protein was significantly increased in the plant extract-treated group. In the experimental group treated with LPS, protein expression was significantly decreased in both experimental groups. Expression was increased in the
(c) Influence on the proliferation of immune cells
In order to examine the effect of the black rice extract (pilot plant extract) prepared according to the above example on the immune cell proliferation effect, the growth promoting effect on the immune cells B and T cells, which play an important role in the human immune system, And evaluated.
B and T cells were incubated for 3 days. The culture was incubated with WST (Water-soluble tetrazolium salt) reagent and absorbance was measured at 450 nm.
As shown in FIG. 5, in the experimental group treated with the pilot extract, the B and T cells did not show the ability to proliferate, and the experimental groups treated with the plant extract showed the ability to proliferate B and T cells (FIG. 5). (* p < 0.05 vs. control)
(d) Immune enhancement effect of NK (Natural Killer) cell
In order to evaluate the immunity enhancement effect of NK cells by the black rice bran extract (pilot plant extract) prepared according to the above example, B cell and T cell supernatant which had been treated with the extract and subcultured four times were added to NK cell culture medium The cells were cultured for 3 days and the proliferative capacity of NK cells was confirmed by WST assay.
As shown in FIG. 6, no significant results were observed in the pilot extract group, and NK cell proliferation was confirmed in the plant extract group. (* p < 0.05 vs. control)
(e) Effect on cytokine expression
In order to investigate the effect of the black rice bran extract (pilot plant extract) prepared according to the above example on the production of cytokine IL-6, the expression level of intracellular mRNA was measured using Real Time-qPCR.
After incubation for 24 hours, the cells were extracted from the cells and subjected to RT-PCR and Real Time-qPCR Respectively.
As shown in FIG. 7, the expression of cytokines was increased in the test group treated with the extracts alone, and in the group treated with LPS, the expression of cytokines increased by LPS was significantly decreased . (* p < 0.05 vs. control, ** p < 0.05 vs. LPS)
Experimental Example 3: Evaluation of immunological activity of cyanidin-3-glucoside (C3G)
In order to evaluate the immunological activity of Cyanidin-3-Glucoside (C3G), a major anthocyanin component of rice black rice, the effects of cytotoxicity, production of NO and H 2 O 2 on macrophages, , Activity on NK cells, and cytokine expression were evaluated.
(a) Effect on RAW 264.7 cell viability
Cytotoxicity of C3G was assessed by MTT assay (cell redox potential) using macrophage RAW 264.7 cells.
The concentrations of the anthocyanins in the black rice extract were compared with those of the anthocyanin extracts of the black rice, and the cells were cultured for 24 hours.
As shown in FIG. 8, C3G showed no toxicity to macrophages up to a concentration of 10 nM.
(b) NO and H 2 O 2 Effect on production
When the immune response is activated, it is known that cells produce NO and H 2 O 2 .
Nitric oxide (NO) is a biologically active molecule with high reactivity and is produced from L-arginine by the iNOS protein. NO is involved in neurotransmission, vascular relaxation, and cell mediated immune responses. Especially, macrophages are stimulated by LPS It is known that iNOS is expressed and produces NO.
Hydrogen peroxide (H 2 O 2 ) is a kind of Reactive Oxygen Species (ROS) that is generated in cells. It is known that when the intracellular inflammatory reaction is increased, it is produced by immune cells and participates in immune reaction with NO. In particular, when ROS such as H 2 O 2 is produced by the immune response, cells express proteins that remove it, and Peroxiredoxin 1 (Prx 1) is known to play a major role.
In order to evaluate the immunoregulatory ability of C3G, the expression of iNOS, a protein that produces NO by inducing an immune response after treatment of macrophages with lipopolysaccharide (LPS), and
The macrophages were pretreated with C3G, treated with LPS, cultured for 24 hours, and the cells were lysed to recover the supernatant. The supernatant proteins were subjected to SDS-PAGE followed by Western blotting to confirm protein expression.
As shown in FIG. 5, when the LPS alone was treated, the immunoregulatory proteins iNOS and
iNOS protein significantly decreased protein expression from 1 nM, the lowest concentration of C3G treatment. The expression of
(c) Influence on the proliferation of immune cells
In order to confirm the effect of C3G on the immune cell proliferation, the growth promoting effect on the immune cells B and T cells, which plays an important role in the human immune system, was evaluated by the WST assay (cell proliferation assay).
B and T cells were added to each concentration of C3G. After culturing for 3 days, the culture was reacted with WST (Water-soluble tetrazolium salt) reagent and absorbance was measured at 450 nm.
As shown in FIG. 6, B cell showed 10-20% proliferative activity when C3G was added at 1 nM, 10 nM and 100 nM, and about 20% . No C3G concentration-dependent proliferative activity was observed in B cells and T cells (Fig. 10). (* p < 0.05 vs. control)
(d) Immune enhancement effect of NK (Natural Killer) cell
NK cells have unique characteristics and are immunoregulatory and immunosuppressive in various immune diseases (autoimmune diseases, infectious diseases, cancer, etc.).
To evaluate the effect of C3G on NK cell immunity, the supernatants of B cells and T cells cultured with C3G for 4 consecutive passages were added to NK cell culture medium and cultured for 3 days. Respectively.
As shown in FIG. 11, the proliferation of NK cells was confirmed in the experimental group treated with C3G at least 10 nM (* p <0.05 vs. control).
(e) Effect on cytokine expression
Cytokine is a protein active substance secreted by lymphocytes, inflammatory cells and hematopoietic cells involved in the immune response. Interleukin (IL) is a substance that is secreted by T lymphocytes and regulates the interaction of other cells.
IL-6 is secreted from monocytes and macrophages. B cells are involved in stimulating differentiation into plasma cells, stimulating secretion of antibodies, inducing the production of Ig, and differentiation and proliferation of T cells.
In order to investigate the effect of C3G on cytokine IL-6 production, intracellular mRNA expression level was measured using Real Time-qPCR.
After C3G treatment of macrophages and LPS treatment for 24 hours, RNA was extracted from the cells and subjected to RT-PCR and Real Time-qPCR.
As shown in FIG. 12, IL-6 mRNA expression level was significantly increased as a result of the LPS-induced immune response, and LPS-induced IL-6 mRNA expression was increased in the C3G-treated experimental group. In the case of C3G treatment, a weak inhibitory reaction was observed at 1 nM and a remarkable inhibitory reaction was observed at 10 nM or higher (Fig. 12). (* p < 0.05 vs. control, ** p < 0.05 vs. LPS)
Experimental Example 4: Evaluation test of immunological activity of red ginseng extract (plant scale) and red ginseng
In order to compare the immunological activity of the black rice bran extract prepared according to the above example, the immunological activity of the extract was compared with that of the red ginseng extract.
The red ginseng extract prepared in this example was dissolved in DMSO for experimental use after drying by spray drying (SD) method. The red ginseng extract was dissolved in commercial DMSO, ug / ml.
(a) Effect on RAW 264.7 cell viability
The cytotoxicity of red ginseng extract was evaluated by MTT assay using macrophage RAW 264.7 cells.
The concentration range was set in comparison with the above experimental example. The extract of black rice rice was not evaluated for toxicity, but the extract was added only for the concentration of red ginseng extract. Cells were cultured for 24 hours and MTT assay was performed.
As shown in FIG. 13, the red ginseng extract showed no toxicity in all the concentration ranges and did not show the same growth ability as the black rice extract.
(b) NO and H 2 O 2 Effect on production
The effects of the black rice bran extracts and the red ginseng extracts prepared according to the above examples on the production of NO and H 2 O 2 related to immunological activity were evaluated.
The expression of iNOS protein was increased by the two extracts when the two extracts were singly treated. In the experimental group treated with LPS, the protein expression was decreased significantly in both experimental groups. Expression was decreased.
As shown in FIG. 14, the expression of
(c) Influence on the proliferation of immune cells
In order to examine the effect of the black rice extract and red ginseng extract on the immune cell proliferation of the B cells and T cells, which play important roles in the human immune system, the WST assay was performed Respectively.
B and T cells were treated with WST (Water-soluble tetrazolium salt) reagent and incubated for 3 days. The absorbance at 450 nm was measured.
As shown in Fig. 15, in the experimental group treated with each extract, B, T cell showed the ability to proliferate. (* p < 0.05 vs. control)
(d) Immune enhancement effect of NK (Natural Killer) cell
To evaluate the immunity enhancement effect of NK cells by the black rice extract and red ginseng extract prepared according to the above examples, B cell and T cell supernatants which had been treated with the extract and cultured for 4 times were added to the culture solution of NK cell After incubation for 3 days, NK cell proliferative capacity was confirmed by WST assay.
As shown in FIG. 16, the addition of B, T cell supernatant showed significant NK cell proliferation in both experimental groups. (* p < 0.05 vs. control)
(e) Effect on cytokine expression
To investigate the effect of the black rice extract and red ginseng extract on cytokine production, the amount of mRNA expression in the cells was measured using Real Time-qPCR.
The cells were treated with the extracts of the black rice extract (Gd) and the red ginseng extract (G) prepared by the above example and the extracts were treated with LPS, and the cells were cultured for 24 hours. RNA was extracted from the cells RT-PCR and Real Time-qPCR were performed.
As shown in FIG. 17, three cytokine expressions were increased by the two extracts in the single treatment group, and all the cytokines were significantly decreased in the LPS-treated group. (* p < 0.05 vs. control, ** p < 0.05 vs. LPS)
Although the present invention has been shown and described with respect to certain preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the following claims It will be apparent to those skilled in the art.
Claims (10)
Centrifuging and filtering the extracted hot-water extract;
Concentrating the filtered extract;
Sterilizing the concentrated extract;
Cooling the sterilized extract; And
And spray-drying the cooled extract to prepare an extract powder.
Wherein the hot water extraction step comprises adding water to the black rice micropowder powder at 50 to 70 캜 and subjecting the resultant to hot water extraction for 1 to 3 hours.
Wherein the step of centrifuging and filtrating is performed by centrifuging with a decanter to remove the extracted foil and filtering the centrifuged liquid from which the extracted foil has been removed with a bag filter .
Wherein the concentrating step is carried out by natural recycling at 50 to 55 ° C. so that the evaporation amount is 500 kg / hr.
Wherein said sterilizing step is sterilized at 90 DEG C for 10 minutes in a hemispherical concentrator.
Wherein the step of cooling is performed at a temperature of 70 캜 or lower.
The step of spray-drying the cooled extract to produce an extract powder may include the steps of: cooling the extracted extract at an inlet temperature of 180 to 200 ° C, an outlet temperature of 80 to 100 ° C, wherein the nozzle press is spray-dried using an electric heater having a pressure of 40 to 70 bar.
which is capable of inhibiting iNOS and Prx 1 protein expression, immune cell proliferation, NK (Natural Killer) cell proliferation, IL-6 mRNA and TNF-alpha cytokine production.
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| KR20190127447A (en) * | 2018-05-04 | 2019-11-13 | 원광대학교산학협력단 | A composition for preventing and treating oral disease comprising black rice |
| KR20240029908A (en) | 2022-08-29 | 2024-03-07 | 농업회사법인 유한회사 별곡 | A Method For Extracting Protein From Rice Bran and Protein From Rice Bran using the same |
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| KR20210058741A (en) * | 2019-11-14 | 2021-05-24 | 대한민국(농촌진흥청장) | Health functional food for ameliorating stress comprising extract of black rice aleurone |
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| JP3653068B2 (en) * | 2002-03-08 | 2005-05-25 | 株式会社エル・エスコーポレーション | Health food and method for producing the same |
| EP1617837A4 (en) * | 2003-04-04 | 2008-08-06 | Tg Biotech Corp | Composition for preventing or treating allergic disease using black rice extract and its therapeutic use |
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| KR101163072B1 (en) | 2009-12-04 | 2012-07-05 | 권진환 | Process for purification and powdering of Aronia berry extract |
| KR20120098070A (en) | 2011-02-28 | 2012-09-05 | 건국대학교 산학협력단 | A subcritical water extraction of black rice bran and antioxidative composition comprising of the same as an effective component |
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| KR20190127447A (en) * | 2018-05-04 | 2019-11-13 | 원광대학교산학협력단 | A composition for preventing and treating oral disease comprising black rice |
| KR20240029908A (en) | 2022-08-29 | 2024-03-07 | 농업회사법인 유한회사 별곡 | A Method For Extracting Protein From Rice Bran and Protein From Rice Bran using the same |
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