KR101253527B1 - Manufacturing method for rehmannia radix preparata - Google Patents

Abstract

The present invention relates to a method that can be produced more economically in a simple process by shortening the maturation period significantly, more specifically, immersed in the sulfur; Primary maturing aged egg yolk for 44 to 76 hours at 70 to 90 ° C. and 80 to 90% humidity; First aged aged sulfur for 45 to 55 ℃, 80 to 90% humidity for 92 to 100 hours secondary aging; And a second step of aged aged second aged sulfur for 44 to 76 hours at 60 to 80 ℃, 80 to 90% humidity; In addition, the present invention relates to sujikuhwang excellent in antioxidant properties and immunity produced by the above method.

Description

Manufacturing method for Sukji sulfur {Manufacturing method for rehmannia radix preparata}

The present invention relates to a method that can be produced more economically in a simple process by significantly shortening the maturation period. In addition, the present invention relates to sujikuhwang excellent in antioxidant properties and immunity produced by the above method.

Rehmannia gluten (Rehmannia glutinosa Libosch.var.purpurea Makino or the root of the same plant) is a perennial herb belonging to the Scrophulariaceae. Lively sulfur is widely recognized for its efficacy in blood, tonic, sedation, anemia, hemostasis, bleeding, uterine bleeding, tuberculosis.

Dried dried turmeric and dried liquor are placed in red wine or white liquor, then steamed, steamed, steamed, and dried repeatedly. They are classified as Rehmanniae Radix Preparata, and their pharmacology is used differently. Sukjihwang has the efficacy of blood, consonant, menstrual irregularities, weak constitution, children's development, dementia, premature ejaculation, erectile dysfunction. In addition, the recent improvement of blood circulation to improve the peripheral microcirculation of various chronic diseases, the increase of blood superoxide dismutase (glucothione peroxidase) and glutathione oxidase (glutathione peroxidase), inhibition of lipid peroxidation, immunosuppressive effect, Increased activity of fibrinolytic system, reduction of polybrene-induced erythrocyte aggregation induced by polybrene, increase of erythrocyte deformability and ATP content have been reported. Sukjihwang is the difference between the pharmacological action of the sulfur and the sulfur is because the steaming, steaming and drying of the sulfur is a new substance beneficial to the human body. The more the steaming and drying process is repeated, the higher the quality of Sukjihwang is recognized. During the steaming process of sucrose, the major saccharide of sucrose is changed to 5-HMF (5-hydroxymethyl-2-furaldehyde). 5-HMF is used as an indicator to evaluate the quality of processed foods with high sugar content. 5-HMF is also used as an indicator for the quality evaluation of ripened sulfur. The higher the content of 5-HMF, the higher the quality of ripened sulfur.

However, in the conventional method for producing sulfur, it is difficult to completely steam even if steamed over several times, so the main needle-steaming-drying must be repeated. Usually steaming 1 ~ 3 days, steaming 1 ~ 3 hours, drying 1 ~ 2 days for 1 steaming, and 2 steaming ~ 9 steaming, the steaming time is reduced to 6 hours ~ 1 day In order to produce Sukji sulfur at a full time, it takes a month or more. There is a problem in that the productivity is very low because the steaming and drying work must be continued in the manufacturing process. In addition, because of the problems of rancidity, germ propagation and multiplication, off-flavor, the alcohol used in the main needle can not be used repeatedly, which leads to higher production costs. As described above, the production process of sulfuric acid is very complicated and requires a lot of time, cost, and manpower. Therefore, it is required to develop a method of producing sulfuric acid, which has excellent physiological activity while being able to manufacture using less raw materials in a shorter time with a simpler process.

In the existing patent on the manufacturing method of the ripening sulfur, charcoal is used in the manufacturing process of the ripened sulfur to make the amount of steam absorbed by the sulfur during the steaming process constant (Registration No. 839326), or adding an active ingredient such as hawthorn or apple Patent Nos. 407237 and 407244 have attempts to enhance physiological activity. However, the above patents basically adhere to the method of vesicle bleeding, and thus the above-mentioned problems are not solved at all.

Registered Patent # 839326 Patent No. 407237 Patent Number 407244

The present invention is to solve the above-mentioned problems, solve the problem of prolonged maturation period, lowering of work efficiency, cost increase due to liquor used in the distillation of the conventional acupuncture, steaming-drying process 9 times It is an object of the present invention to provide a method that can be produced economically by a simple process in a short period of time and the sulfur content prepared by the method.

It is another object of the present invention to provide a method for producing sucrose which is excellent in physiological activity such as sulfated activity and immune activity after ripening, and sucrose which is prepared by the method.

The present invention for achieving the above object is a step of immersing in sulfur; Primary maturing aged egg yolk for 44 to 76 hours at 70 to 90 ° C. and 80 to 90% humidity; First aged aged sulfur for 45 to 55 ℃, 80 to 90% humidity for 92 to 100 hours secondary aging; And a second step of aged aged second aged sulfur for 44 to 76 hours at 60 to 80 ℃, 80 to 90% humidity;

When the aging step is completed may further include the step of drying after the third aging so that it can be used for distribution and storage without altering for a long time. Alternatively, it can be used after being refrigerated or refrigerated when used for a short time. The drying process may be carried out under the conditions used in the drying of common sulfur. That is, it may be dried at room temperature or may be dried by hot air drying at 50 ~ 80 ℃ hot air. The temperature of the hot air is more preferably 50 ~ 65 ℃ in order to prevent the carbonation of sulfur and the production of benzopyrene, which is a carcinogen during drying. The drying time depends on the size of the drying equipment, the drying temperature, Naturally, it should be appropriately adjusted according to the amount of sulfur to be dried together.

In the following, the optimum conditions of the manufacturing process of the sulfur of the present invention will be described in more detail with reference to the preliminary experimental results.

In the present invention, the turmeric is characterized in that it is raw or dried sulfur. Even in the prior art, the ripened sulfur is prepared from raw sulfur or dried sulfur, the present invention can also produce the ripened sulfur from raw sulfur or dried sulfur.

Jujum is to immerse the sulfur in immersed in liquor, the liquor can be used as a sulfur or white wine, considering the economic aspects it is preferable to immerse in sulfur. Both Hwangju and Liquor are grain wines, and Hwangju is yellow like Makgeolli, and Liquor means clear and transparent wine. The bitter taste of jihwang is removed during the immersion process, and the sugar content of sujijihwang is increased by sugars contained in grain wine, white wine, and white wine.

Since dried liquor removes most of the moisture, white liquor or yellow wine easily penetrates the liquor. Therefore, the effect of distillation is more effective even if the dipping time is shorter than the raw yellow liquor, which has to be replaced with white liquor or yellow liquor. Excellent. In other words, when raw sulfur is used as a raw material, it is preferable that the main needle is made in the range of 48 to 72 hours, and when the dry sulfur is used as a raw material, it is sufficient to immerse it for 24 to 48 hours. Therefore, it is preferable to perform a lap needle for 24 to 76 hours at normal temperature. In addition, it is preferable to spray the liquor used during the aging during the first and third ripening step. The color of saengjihwang is darkened by sprinkling and the gloss is formed on the surface to improve the color and sugar content. The aging step means just before aging, middle or immediately after aging. Because the secondary ripening step is low temperature, if the sprinkling of alcohol in the second ripening step may cause mold due to the yeast of the liquor. Therefore, the sprinkling of alcohol is limited to the first and / or third ripening step, and the number of times that the sprinkling of alcohol in the ripening step is preferably 1 to 4 times. If the number of sprinkles is too high, the process efficiency is reduced.

The first aging step is carried out 44 ~ 76 hours at 70 ~ 90 ℃, 80 ~ 90% humidity. If the temperature is less than 70 ℃ mold caused by the yeast of the liquor used in the alcohol or a lot of smell peculiar to the liquor, if the temperature is too high, benzopyrene (benzo (α) pyrene) is produced. Benzopyrene is a polycyclic aromatic hydrocarbon compound that is mainly produced during the non-combustion process of organic compounds. Benzopyrene is known to be toxic to the liver by oxidizing it into cytochrome P-450 to form 7-8-diol and then reoxidizing to diepoxide. When the humidity is more than 90% of the ripening, turmeric decayed, less than 80% of the maturation effect was not good. In addition, if the maturation time is less than 44 hours, the maturation of the sulfur was low, and if the ripening for more than 76 hours, the surface was browned without the maturation of the maturity or the maturation of the inside.

When the first ripening step is completed, lower the temperature to 45 ~ 55 ℃, 92 ~ 100 hours Secondary aging. The secondary aging step is to allow the aging to be sufficiently done to the inside while preventing the browning change or burning of the surface by continuing aging in milder conditions. In the second aging step, if the aging temperature is too low or the aging time is too short, aging did not occur sufficiently. If the temperature is too high, browning will occur in a state where the inside is not sufficiently aged. Secondary aging time is 100 hours is enough, and a slight increase in time does not lower the quality of the turmeric, but in consideration of economical efficiency is preferably less than 100 hours.

When the second ripening is completed, to reduce the sour taste of the ripening sulfur and to increase the sugar content, and to make the surface brown, to increase the color, the ripening temperature is raised to 60 ~ 90 ℃ again for the third aging 44-76 hours. If the temperature is too low or the time is too short, the acidity of sujijihwang high acidity is strong, while the sugar content is low and the color is not good. On the other hand, if the temperature is too high or the time is too long, benzopyrene may be formed and the surface may burn. Naturally, the lower the aging temperature, the longer the aging time should be.

According to the method of producing sukji sulfur according to the present invention consisting of a total of three aging step as described above, 8 to 13 days, preferably 9 to 11 days including the immersion time can be produced sujiji sulfur because it takes more than a full time early Significantly reduced time can greatly improve productivity. In addition, since only one time the needle is used, the amount of alcohol used in the needle is only 10-20% of the conventional production cost is significantly lower.

The first to third aging step is more preferably carried out in a aging machine equipped with a constant temperature, humidity. In the case of using a aging machine equipped with a constant temperature and humidity function, the temperature and humidity can be adjusted more easily, thereby further improving productivity.

The present invention also relates to sulphurous sulfur produced by the above method.

According to the manufacturing method of the present invention, not only the time required for producing the ripened sulfur is short, but also the yield compared to the dry weight of the ripened sulfur by the method of dry pulp, which can be produced more economically. Sulfur sulfur according to the present invention is significantly improved in the sugar content by the conventional method and the pH is not low, so the texture is more excellent. In addition, the content of 5-HMF, which is an indicator of Sukji sulfur, was significantly higher than that of Sukju, and the content of Catapol, which causes gastrointestinal disorders, was much lower than that of Sukju. In terms of antioxidant activity, all the items of DPPH radical scavenging ability, FRAP value, Hydroxy radical scavenging activity and total phenolic content, which are the measurement criteria, showed superior efficacy compared to succinic sulphur. In addition, the cytotoxicity of the RAW264.7 cell line, the ability to inhibit NO production, and the cytokine production were lower than those of S. aureus.

As described above, the method of manufacturing sucrose sulfur of the present invention greatly shortens the manufacturing period of sucrose sulfur, which takes 15 to 30 days or more, and the process is very simple, and greatly reduces the amount of liquor used for sake, improving productivity and producing The unit price can be greatly reduced.

In addition, according to the present invention, by using a aging machine that maintains a constant temperature and humidity, the temperature and humidity are maintained without opening and closing the aging machine, and it is possible to solve the inconvenience of having to repeat the conventional needle-vapor-drying nine times.

Figure 1 is a photo of sukji sulfur produced by one embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these embodiments are merely examples for explaining the content and scope of the technical idea of the present invention, and thus the technical scope of the present invention is not limited or changed. In addition, it will be apparent to those skilled in the art that various modifications and changes can be made within the scope of the present invention based on these examples.

Example

The raw sulfur used in the preparation examples and the comparative examples was considered to be Goryeo's sulfur, which was used by purchasing GAP sulfur grown in a farmer's house in Namsan, Geumsan-gun. Guernsey sulfur was used by drying the purchased Goryeo sulfur in 3 ~ 4 days in a dryer of 40 ~ 50 ℃.

Preparation Example 1 Preparation of Sulfur Sulfur from Raw Sulfur

The raw paprika was cut into 3 cm lengths, and the rice wine was soaked (alcohol degree 7 °) so that the raw paprika was submerged for 48 hours at room temperature. After picking out the dried raw sulfur, put it in a tray, put it in a thermo-hygrostat, and aged it for 48 hours at 80 ° C and 80-90% humidity. After 24 hours and 48 hours after the first aging, 50 ml of rice wine was sprayed per 100 g of fresh sulfur. When the first ripening was completed, the internal temperature of the thermo-hygrostat was lowered to 50 ° C., and the second ripened for 96 hours at a humidity of 80 to 90%. When the second ripening was completed again, 50 ml of rice wine was sprayed per 100 g of fresh sulfur, and the internal temperature of the thermo-hygrostat was raised to 80 ° C. to ripen for three hours at 80 ° C. and 80 to 90% humidity for 48 hours. The prepared sulfur content is shown in Figure 1 (a).

Preparation Example 2 Preparation of Succinate Sulfur from Dried Yellow Sulfur

Dried guangji was cut into 3 cm in length and put the makgeolli (alcohol water 7 °) enough to immerse the dried guangji was 48 hours at room temperature. After dipping the dried Guernsey sulfur, put it in a tray, put it in a thermo-hygrostat, and aged it first at 70 ° C. and 80-90% humidity for 72 hours. After the first aging, 50 ml of makgeolli was sprayed per 100 g of dried goji berries. When the first ripening was completed, the internal temperature of the thermo-hygrostat was lowered to 50 ° C., and the second ripened for 96 hours at a humidity of 80 to 90%. When the second ripening is completed, spray 50 ml of rice wine per 100g of Guernsey Sulfur, raise the internal temperature of the thermo-hygrostat, and then 24 hours at 70 ℃, 80 ~ 90% humidity, then 48 at 60 ~ 70 ℃, 80 ~ 90% humidity. Sulfur was prepared by aging three times. Photographs of the prepared sulphur was shown in Figure 1 (b).

Comparative Example 1 Preparation of Ripe Yellow Sulfur by Gutyocysis of Raw Sulfur

Raw paprika was cut into 3 cm and put soaked rice wine soaked for 48 hours at room temperature. The steamed raw sulfur was steamed for 2 hours, and hot air dried at 50 ° C. for 18 hours to prepare 1 steamed sulfur. From 2 steam to 9 steam, except for 6 hours of immersion in makgeolli, the steaming, steaming, and drying process were repeated up to 9 steam by the same method as 1 steaming to prepare hungjeokpo sujiji sulfur.

Comparative Example 2 Preparation of Sukjihwang by Gujeobu Bubbles

Except for using dry sulfur instead of raw sulfur as a raw material was prepared jingugu sukji sulfur by the same method as in Comparative Example 1.

Example 1 Measurement of Production Yield and Water Content of Sulfur

Table 1 shows the production yield of the ripened sulfur of the above Preparation Example and Comparative Example by calculating the amount (%) of the final ripening sulfur obtained from 100 g of raw material. For accurate comparison of the yield, raw moisture of dried raw and dried sulfur and dried sulfur were measured and shown together. Moisture was measured using an infrared moisture meter (Sartorius, Germany) after taking 2-3 g of the sample by grinding the green, dry or sulfur. For comparison March 2010 purchased on the market The moisture content of the commercial ripened sulfur was shown together.

As can be seen from Table 1, the production examples 1 and 2 of the present invention compared to the method of Comparative Examples 1 and 2 according to the prior art when comparing the yield calculated in terms of dry weight when using raw sulfur as a raw material It was 5.9 times higher and 7.2 times higher than dried Guangji sulfur. Particularly, in the case of Preparation Example 1 using raw sulfur, the yield was 142.2% of the dry weight, and the yield was more than 100%. This is because Makgeolli was replaced with moisture and the useful components of Makgeolli were accumulated in the riped sulfur. .

The low yield of sulphurous sulfur in the comparative example due to vesicles is thought to be due to the dissolution of the water-soluble components of the sulfur in the repeated immersion process, and the yield reduction as the immersion sulfur in the preparation example of the present invention is omitted. It is judged that did not occur. The decrease in yield means not only the amount of sucrose which was obtained, but also that the loss of useful components did not occur during the manufacturing process, and thus, the content of useful components in the prepared sucrose was expected to increase greatly.

Example 2 Evaluation of Physicochemical Properties of Sulfur

After preparing the sample by grinding the sujijihwang prepared in the Preparation Example and Comparative Example, the results were examined in the physicochemical properties of sugar and pH are shown in Table 2 below. For comparison March 2010 purchased on the market It also shows the characteristics of commercially ripened sulfur.

pH was measured according to the Association of Official Agricultural Chemists (AOAC) method (1990). That is, 5 g of the sample was put together with 20 mL of distilled water, homogenized, and centrifuged at 3,000 rpm for 15 minutes. pH was measured with a supernatant pH meter (420 Benchtop, Orion Research, USA).

The sugar content was taken from the supernatant and measured with a sugar meter (N-1E Brix 0-32%, Atago, Japan).

The ripening sulfur of Preparation Example 1 and Preparation Example 2 prepared by the manufacturing process of the present invention showed a higher sugar content than that of Comparative Example 1 and Comparative Example 2 prepared by the agglomerate cells of the prior art, and the pH was similar or Slightly higher. Commercially available sulfur content was low in both sugar and pH.

Example 3 Analysis of Active Ingredient Content of Sulfur

Reducing sugar, Catapol, 5-HMF content in Preparation Examples 1 and 2, Comparative Examples 1 and 2, and commercially available Sukji sulfur were measured, and the results are shown in Tables 3 and 4, respectively. Reducing sugar content was expressed as glucose content by measuring the absorbance at 550 nm using a spectrophotometer (Model UV-1800, 240V) by colorimetric method by DNS using the supernatant used in the pH and sugar measurement of Example 1. The standard curve was prepared by reacting glucose (Duksan pharmaceutical Co., LTD. Yonginuoop, Kyongkido, Korea.) According to the concentration.

The content of Catapol and 5-HMF was 2 g of sucrose and sample, 100 mL of 30% MeOH was added and blended for 1 minute. After extracting by filtration with ultrasonic shaking for 2 hours, the filtrate was centrifuged at 13000 rpm for 10 minutes. The supernatant was filtered with 0.45 μm syringe fliter and analyzed by HPLC. Mobile phase conditions were 1% ACN, flow rate was 1 min / mL, and HPLC column was Agilent (USA), C18 4.6 × 150 nm 5 ㎛. The sample injection volume was 10 μl, and the analysis wavelength was measured at catalpol 204-206 nm and 5-HMF 285 nm.

As can be seen in Table 3, the reducing sugar content of the preparation of sujijihwang prepared by the manufacturing method of the present invention is about 1.15 times when prepared from raw suhwanghu, and about 1.5 times higher when prepared from dried sujihwang compared to the comparative example sujihwang by the gujeunggu The value is shown. This indicates that the reducing sugar content is increased even if the number of spraying makgeolli in the dipping and ripening process of makgeolli in the method of the present invention decreases.

Referring to Table 4, catapol, which is known to cause digestive disorders, which is a representative side effect of turmeric, contained only 6.7% of Preparation Example 1 compared to Comparative Example 1, and Preparation Example 2 was not detected. On the contrary, the 5-HMF content, which is an indicator material of the ripened sulfur, was increased about 3.2 times compared to Comparative Example 1, and the ripened sulfur of Preparation Example 2 was 23 to 25 times higher than that of Comparative Example 1 or commercially available ripened sulfur by vesicles. Increased. The 5-HMF content of commercially ripened sulfur was almost similar to that of Comparative Example 1.

Example 4 Analysis of Antioxidant Activity of Sulfur

50 mL of methanol was added to 1.5 g of the powdered dry sample of Preparation Example or Comparative Example, homogenized, and stirred at room temperature for 15 hours. Remove the methanol solution of 4 ℃, 20 minutes at 3000 rpm centrifugation by concentration under reduced pressure using an evaporator and the resulting supernatant to give the extract of 0 .6 ~ 0.7g each. Antioxidant activity was measured by the following method using the obtained extract, and the result is shown in Table 5.

1) Total Phenolic Content Analysis

Samples were prepared by adding 20 mM PBS buffer so that the final concentration of the extract was 250 mg / mL. In 2.5 mL of distilled water, 0.33 mL of sample, 0.16 mL of Foline-Denis reagent, and 0.3 mL of saturated Na ₂ CO ₃ solution were developed for 30 minutes in the dark, and the absorbance was measured at 760 nm to quantify the content of phenol. (Standard used tannic acid.)

2) DPPH radical scavenging activity measurement

Methanol was added to the final concentration of the extract to 250 mg / mL was used as a sample solution. 150 μL of a 1.5 × 10 ⁻⁴ mM DPPH (1,1-diphenyl-2-picryl hydrazyl) solution was added to 50 μL of the sample solution, and the absorbance was measured at 515 nm using a spectrophotometer after 30 minutes. After calculating the radical scavenging ability (%) by the following equation (1), IC ₅₀ which is a concentration at which the radical scavenging ability becomes 50% from the calibration curve for the radical scavenging ability of each concentration was obtained.

&Quot; (1) "

3) Hydroxy radical scavenging activity measurement

20 mM PBS buffer was added to the final concentration of the extract to 250 mg / mL was used as a sample solution. To 0.15 mL of sample solution, add 0.35 mL of buffer, 0.1 mL of 3 mM deoxyribose solution, 0.1 mL of 0.1 mM ascorbic acid solution, 0.1 mL of 0.1 mM EDTA solution, 0.1 mL of 0.1 mM FeCl ₃ solution, and 0.1 mL of 1 mM H ₂ O ₂ solution. After stirring well, the reaction was carried out at 37 ° C. for 1 hour. After the reaction, 1 mL of 2% TCA solution and 1 mL of 1% TBA solution Put them in order and mix and react for 20 minutes at 100 ℃ Cooled and centrifuged. The supernatant obtained by centrifugation was measured for absorbance at 532 nm using a spectrophotometer, the radical scavenging ability (%) was calculated by Equation 2 below, and the radical scavenging ability was 50% in the calibration curve for each concentration. IC ₅₀ , which is a concentration at, was obtained.

&Quot; (2) "

3) Ferric-reducing antioxidant potential (FRAP) measurement

FRAP measurements were made with reference to the methods of Benzie IFF and Strain JJ (Methods Enzymol. 299: 15-27, 1996). FRAP samples were prepared by warming 25 mL acetate buffer (300 mM, pH3.6) at 37 ° C and then dissolving 10 mM 2,4,6-Tris (2-pyridyl) -s-tri-azine (TPTZ) dissolved in 40 mM HCl. , Sigma Chemical co., St. Louis, MO, USA) 2.5 mL and 20 mM ferric chloride (FeCl ₃ ) 2.5 mL was added thereto. 0.03 mL of the extract and 0.09 mL of distilled water were added to the prepared 0.9 mL FRAP reagent and reacted at 37 ° C. for 10 minutes, and the absorbance was measured at 593 nm. From the measured absorbance, the FRAP value was obtained by the following equation (3). The control group was measured by adding ethanol instead of the sample. The standard curve was calculated by substituting the calibration formula of FeSO ₄ , which was repeatedly prepared at concentrations of 0.025, 0.05, 0.1, 0.2, 0.5 and 1 mM.

&Quot; (3) "

In this case, x means FRAP value and y means absorbance of 593 nm at the corresponding concentration.

From Table 5, the sulfur content of the preparation example was confirmed that the IC ₅₀ value for radical scavenging decreased compared with the comparative example according to the prior art. IC ₅₀ for DPPH radical scavenging was significantly reduced to 56.3% in Preparation Example 1 and 20.1% in Comparative Example 2 compared to Comparative Example 1, and IC ₅₀ for Hydroxyl radical scavenging was prepared compared to Comparative Example 1. Example 1 was reduced to 98.6%, Preparation Example 2 was 83.3% compared to Comparative Example 2. The FRAP value of Manufactured 1 and 2 increased 109% and 308.4%, respectively, compared to Comparative Examples 1 and 2, and the total phenolic content also increased by 150% and 215%. Indicated. In particular, the antioxidant capacity of the ripening sulfur of Preparation Example 2 prepared from dried koji sulfur was the best.

Example 5 Analysis of Cytotoxicity and Immune Ability of Sukjihwang

1) Analysis of Immunity in RAW 264.7

(1) Preparation of raw or sulfur

Sulfur sulfur sample or saenghwang sulfur sample of Preparation Example 1, Comparative Example 1 or Comparative Example 2 was lyophilized and then ground to a powder. 5 g of powder was added to 45 mL of distilled water and shaken with ultrasonication for 2 hours. After centrifugation at 3000rpm for 15 minutes, only the supernatant was collected and concentrated under reduced pressure using an evaporator, and distilled water was added so that the final concentration was 1 g / mL.

(2) RAW 264.7 cell line culture

The mouse macrophage cell line RAW 264.7 was distributed by the Bank of Korea Cell Line and 10% heat-inactivated fetal bovine serum (10% FBS) and 1% penicillin in Dulbecco's Modifide Eagle's Medium (DMEM, Hyclone Laboratories, Inc. cat.SH30243.01). The solution was added to a 100 mm dish and incubated at 37 ° C. in a 5% CO ₂ incubator.

The number of macrophages was adjusted to 5 × 10 ⁴ cells / mL cells and the cells were dispensed into each well of a 96 well culture plate and incubated for 9 hours in 37 ° C. and 5% CO ₂ incubator. After cultivation, Preparation Example 1, Comparative Example 1, or a sample of Live Broccoli was added to each well according to the concentrations shown in Table 5 below, and 2 ug / mL of lipopolysaccharide (LPS from E. coli 055: B5, SIGMA cat.M2128) was added thereto. Was adjusted to 100 μL and then cultured in 37 ℃, 5% CO ₂ incubator. After 22 hours, 10 hours of thiazolyl blue tetrazolium bromide (<= 97.5% TLC, SIGMA, M5655-1G, 53096D) reagent for measuring lymphocyte proliferation capacity was added so that the time of LPS treatment was 24 hours. Incubated further in a% CO ₂ incubator.

(3) RAW 264.7 proliferation measurement

MTT assay was carried out to investigate the cytotoxic effects of Sook-ji-hwang or Saeng-hwang-hwang according to the preparation method in RAW 264.7 cells. After removing the culture medium from the culture cells of (2), the culture cell surface was washed with phosphate buffered saline (1 × PBS). 100 μL of DMSO (SIGMA, cat.D2650, Lot.BNBB3566) was added, and after 10 minutes, cell proliferation was measured by absorbance at 540 nm, and the results are shown in Table 6.

Referring to Table 6, while the concentration-dependent toxicity to cell proliferation was shown, Comparative Example 1 showed more than 95% cell proliferation ability up to 2.5 mg / mL concentration, and Preparation Example 1 showed more than 95% cell proliferation capacity at all test concentrations. Indicated.

(4) NO production inhibitory ability

The amount of nitric oxide (NO) produced from RAW 264.7 cell line was measured using the nitric oxide detection kit (iNtRON, Lot.90510243, Cat.21021) as the form of NO ₂ present in the cell culture. Indicated. More specifically, 100 μL of the cell culture supernatant of (2) and 100 μL of Griess reagent (1% sulfanilamide in 5% phosphoric acid + 1% a-naphthylamide in H ₂ O) were mixed and reacted for 10 minutes in a 96 well plate. After absorbance at 540 nm was measured. The concentration of NO ₂ was calculated by a standard curve made using sodium nitrite.

The NO production of LPS-treated spheres was 11.5 μM, resulting in about 2.9-fold increase of NO production by LPS treatment. The raw sulfur and the homework sulfur of Comparative Example 1 and Preparation Example 1 all reduced NO production in a concentration-dependent manner, but the raw sulfur was the lowest in the reduction rate and the reduction of NO production by the ripe sulfur of the production example was most remarkable.

(5) TNF-α Secretion Capacity of RAW 264.7 Cells

The culture supernatant of RAW 264.7 was obtained by centrifugation and then in supernatant using TNF-α (cat.88-7324-88) and IL-6 (cat.88-7064-88) ELISA set (eBioscience immunoassy kit). TNF-α and IL-6 (cat.88-7064-88) concentrations were measured quantitatively and the results are shown in Table 8 and Table 9, respectively.

Referring to Table 8 and Table 9, both the raw sulfur and the ripening sulfur of Comparative Example 1 and Preparation Example 1 inhibited the secretion of TNF-α and IL-6, as well as the inhibition of NO production, the effect of raw sulfur is the lowest, Was most noticeable. Particularly, for the TNF-α secretion, the ripening sulfur of Preparation Example 1 showed a strong inhibitory effect compared to the raw sulfur or the ripening sulfur of Comparative Example 1.

Claims (6)

Digging the tongue;
Primary maturing aged egg yolk for 44 to 76 hours at 70 to 90 ° C. and 80 to 90% humidity;
First aged aged sulfur for 45 to 55 ℃, 80 to 90% humidity for 92 to 100 hours secondary aging; And
Second aged aged sulfur for 44-76 hours at 60-80 ℃, 80-90% humidity;
Sukji sulfur production method characterized in that the sprinkling of the alcohol used in the first hand and the aging step in the third aging step.
The method of claim 1,
Step of drying after the third aging; The manufacturing method of sukji sulfur further comprising a.
3. The method according to claim 1 or 2,
The sulfuric acid is a production method of ripe sulfur, characterized in that the raw sulfur or dried sulfur.
delete 3. The method according to claim 1 or 2,
The first to third aging step is a method of producing sukji sulfur characterized in that it is carried out in a aging machine equipped with a constant temperature, humidity.
Sukji sulfur produced by the method of claim 1 or 2.

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