KR101043807B1 - A functional salt and a method of preparing the same - Google Patents

Abstract

The present invention relates to a functional salt and a method for preparing the same. In particular, the functional salt produced by the production method of the present invention is remarkably excellent in antioxidant activity compared to the existing salt, has an excellent free radical scavenging activity, excellent liver protection function, as well as natural substances as an active ingredient As it does not cause resistance or safety related problems, it can be widely used for liver protection and liver damage prevention.

Sun salt, antioxidant, liver

Description

Functional salt and preparation method thereof {A FUNCTIONAL SALT AND A METHOD OF PREPARING THE SAME}

The present invention relates to a functional salt and a method for preparing the same.

The main component of salt is sodium chloride (NaCl), which is physiologically indispensable for animals. Specifically, sodium chloride is contained in the body, in particular, body fluids to adjust the osmotic pressure of the body fluids, sodium as a component of sodium chloride is combined with phosphoric acid to maintain the acid group parallel to the body fluid as a buffer material, such as bile juice, interest solution or serous It forms an alkaline digestive fluid, acts as a neurostimulant, and chlorine, another component of sodium chloride, constitutes gastric juice.

Salt used as a seasoning to taste the food in daily life is a component that is closely related to the physiological activity and life support of the animal as described above, so if the salt intake is short-term, the secretion of digestive fluid is insufficient Loss of appetite occurs, and in the long term, general force, boredom, fatigue or mental anxiety occur, and sudden loss of salinity in the body causes obvious physical and mental loss such as dizziness, indifference, conscious turbidity, and deprivation.

On the other hand, the liver is the largest organ in our body is a very important organ responsible for various metabolism, detoxification, decomposition, synthesis and secretion. More specifically, it has the function of managing energy metabolism, so that all the nutrients absorbed from food are metabolized to the liver to produce energy and supplied or stored throughout the body, and serum proteins of about 2,000 enzymes, albumin and coagulation factors Fats such as bile acids, phospholipids, and cholesterol are synthesized, stored, and distributed in the liver. In addition, since the liver detoxifies drugs, alcohol, toxic substances and the like as a detoxifying and degrading function, hepatocytes are susceptible to damage and therefore drug, toxic, alcoholic liver diseases, and the like can frequently occur. The liver also plays an important role in maintaining our lives because it has the ability to excrete various metabolites into the duodenum through the bile ducts and immune function.

As described above, the liver functions to protect the whole body, and the risk of exposure to many toxic substances in addition to nutrients is higher than that of other organs, which is very high.

The liver is an organ with excellent regenerative capacity, and if there is a slight damage, the liver recovers to a normal level.However, if the damage persists, part of the liver tissue is completely destroyed and liver function is reduced. . When these liver damage becomes chronic, liver fibrosis or cirrhosis of the liver, regardless of the cause progresses to liver cancer. Liver fibrosis, cirrhosis, liver cancer, etc. are currently a disease that does not have a clear treatment and treatment, the mortality rate is also a disease. Therefore, preventing and treating such liver damage before it becomes chronic is very important to inhibit the progression of liver fibrosis, cirrhosis or liver cancer.

The liver has no subjective symptoms at the beginning of the disease, and only when the degree of liver damage is severe, subjective symptoms appear, so early detection of liver disease is difficult.

Currently, the most commonly used drugs for the prevention and treatment of hepatitis and cirrhosis include ursodeoxycholine acid, immunostimulants, antiviral agents, silymarin, unzidose polysaccharides, vitamin B group, and methionine. There has been no clinically satisfactory effect yet. Therefore, in recent years, efforts have been made to find a way to prevent as well as to treat liver damage without burdening the liver easily in usual times.

Therefore, in relation to the prevention of liver function, it has an antioxidant effect, can prevent liver damage caused by oxidative stress, protect liver cells, protect the liver from natural substances that have no problems with resistance or stability and It is urgent to develop an ingredient effective for preventing liver damage and a functional food composition using the same as an active ingredient. In particular, the three sides are the sea, in consideration of the geographical environment of Korea, which is rich in marine resources, there is an increasing demand to develop functional food compositions effective in protecting liver from liver natural substances and preventing liver damage.

The present invention is to improve the problems of the prior art as described above, to provide a functional food composition which is effective in preventing liver damage and preventing liver damage comprising a natural substance as an active ingredient and a method for improving the antioxidant capacity of the natural substance. For the purpose of

In order to achieve the above object, the present invention provides a method for producing sun salt with improved antioxidant capacity and a sun salt with excellent antioxidant capacity prepared by the method.

In addition, in order to achieve the above object, the present invention provides a functional food additive composition effective for liver protection and liver damage prevention comprising the functional salt as an active ingredient.

The inventors of the present invention while studying antioxidants derived from natural substances that do not have a problem of stability, such as side effects or resistance, when salt and bamboo are mixed at a specific ratio and heated by a specific method, the antioxidant activity of the salt is remarkably The present invention was completed, confirming that the hepatoprotective function was improved.

Hereinafter, the present invention will be described in more detail.

In the present invention, salt means all types of salt that can be used for food, not for industrial purposes, and include salt, rock salt, flower salt, roasted salt, bamboo salt and refined salt or refined salt, molten salt, refined salt, processed salt, and the like. And, preferably, sun salt or purified salt.

In the present invention, the natural salt refers to an unprocessed salt made by concentrating seawater by evaporating water using natural forces such as wind and sunlight after introducing seawater or seawater into a reservoir or salt field, and having coarse and translucent hexagons. Has the characteristic of having a crystal. The sun salt is called mineral salt because it is rich in many minerals (minerals) such as calcium, magnesium, zinc, potassium, and iron.

The present invention provides a method for preparing salt having improved liver protection and improved antioxidant activity.

The method for preparing salt having improved hepatoprotective function and improved antioxidant capacity includes: a) a first heat treatment step of heat-treating a natural salt; b) mixing the heat-treated natural salt and bamboo; c) a second heat treatment step of performing heat treatment while raising the temperature of the heat-treated natural salt and bamboo mixture, and d) a third heat treatment step of maintaining the temperature of the mixture on which the second heat treatment step is performed.

In the present invention, the natural salt refers to an unprocessed salt made by concentrating seawater by evaporating water using natural forces such as wind and sunlight after introducing seawater or seawater into a reservoir or salt field, and having coarse and translucent hexagons. Has the characteristic of having a crystal.

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The first heat treatment step a) of heat-treating the natural salt is performed for 30 minutes to 24 hours, preferably 1 hour to 12 hours, more preferably 2 hours to 700 hours, at 700 to 900 ° C, preferably 750 to 850 ° C. The heat treatment may be performed for 6 hours, and the heat treatment may be performed once.

The bamboo is a generic name of an evergreen perennial plant belonging to a monocotyledonous plant rice plant and bamboo subfamily, preferably, the maturity of bamboo may be three to four years old. The bamboo is preferably cut off and used immediately because the sap is insufficient when one week or more. For example, the bamboo may be to use the form of a chip (chip) after crushing the bamboo maturity of 3 to 4 years old. The bamboo chip may have a size of 1 cm × 2 cm, in consideration of the degree of mixing with salt, productivity and process cost.

B) mixing the heat-treated natural salt and bamboo is performed by mixing the natural salt and bamboo at a weight ratio of 40:60 to 70:30 (salt: bamboo), preferably 50:50 to 60:40. can do.

In the case of mixing the natural salt and bamboo, it is remarkably superior to the mixture of bamboo and MgSO ₄ , which are added to prepare the salt, and a mixture of Banbusae Caulis and MgSO ₄ together. It was confirmed that there is an antioxidant activity improving effect.

C) heat-treating while raising the temperature of the heat-treated natural salt and bamboo mixture is heated to the salt and bamboo mixture for 4 to 6 hours, preferably 4 hours 30 minutes to 5 hours 30 minutes so that the mixture is 1,100 ℃ By heat treatment until the The second heat treatment process may be performed by the method described in FIG. 1 as an example. Specifically, the salt and bamboo mixture is heated so that the temperature rises to 2 ° C. per minute until the temperature reaches 200 ° C., and the heat is increased to 4 ° C. per minute until the temperature reaches 200 ° C. to 400 ° C., and 400 ° C. to 1,100 ° C. This can be done by applying heat so that the temperature rises up to 4 ° C. per minute. More specifically, in the above method, in order for the salt and bamboo mixture to be heated from 0 ° C. to 200 ° C., 1 hour 20 minutes to 2 hours may elapse, and the mixture is heated from 200 ° C. to 400 ° C. In order to be, 30 minutes to 1 hour 10 minutes may be elapsed, and in order for the mixture to be heated from 400 ° C. to 1,100 ° C., 2 hours 30 minutes to 3 hours 10 minutes may be elapsed.

The step d) of performing heat treatment while maintaining the temperature of the mixture in which the second heat treatment step is performed may be performed for 15 to 60 minutes, preferably 20 to 40 minutes, at 1,100 ° C. for the mixture. This can be done in a way. Step d) as the third heat treatment process may be performed continuously with step c).

In the case of performing the heat treatment process, unlike the conventional method for producing bamboo salt, not only can produce a functional salt with a significant antioxidant capacity in one heating process, but also heated to 1,200 to 1,300 ℃ to dissolve the salt Compared with the existing processes such as re-solidification and crushing, the work process is simple and the manufacturing cost such as fuel cost and labor cost is reduced.

The present invention also provides a salt which is improved in the hepatoprotective function produced by the production method, and the antioxidant activity is improved. The salt with improved antioxidant capacity prepared by the preparation method was found to be very excellent in the antioxidant activity in the experiment to measure the antioxidant activity using DPPH, the animal experiment was confirmed to have excellent hepatoprotective activity.

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In addition, the present invention provides a functional food additive composition which is effective in preventing liver damage and preventing liver damage, which comprises a salt having improved liver protection function and an antioxidant activity as an active ingredient prepared by the method.

Improved hepatoprotective function produced by the above method, and the improved antioxidant capacity of the salt was measured by various methods, it was confirmed that the remarkably excellent antioxidant capacity, hepatocyte protection function is excellent, there is no cytotoxicity It has been proposed as an active ingredient of a functional food additive composition that is effective in protecting liver, preventing liver damage.

The present invention relates to a functional food additive composition which is effective in preventing liver damage and preventing liver damage, comprising a salt having improved liver protection function and an antioxidant activity as an active ingredient prepared by the manufacturing method. Improved liver protection function prepared by the manufacturing method, food additive composition which is effective in preventing liver damage and liver damage containing salts with improved antioxidant capacity as an active ingredient while adding the salty taste of foods such as tablet salt or rock salt Its excellent antioxidant activity prevents and improves liver damage caused by oxidative stress. Therefore, the food additive composition of the present invention, which can be used in place of tablet salt or rock salt added to give saltiness to food, improves the taste of food and at the same time is effective in protecting liver and preventing liver damage, thereby improving diseases related to liver damage. Or an excellent effect of preventing.

In the present specification, the term "food" means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. It is intended to include all foods, health foods and beverages.

As a food to which the functional food additive composition which is effective in the liver protection and liver damage prevention which contains the salt which improves the hepatoprotective function manufactured by the said manufacturing method of this invention, and improves antioxidant activity as an active ingredient can be added, For example, there are various foods, beverages, gums, teas, vitamin complexes, functional foods, and the like. In addition, the food in the present invention includes special nutritional products (e.g., prepared oils, infants, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasoned foods ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, and the like, and natural seasonings (eg, ramen soup). The food, beverage or food additives may be prepared by a conventional manufacturing method.

The functional food additive composition, which is effective in preventing liver damage and preventing liver damage, including a salt having improved liver protection function and an antioxidant activity as an active ingredient, prepared by the manufacturing method, various ingredients usually included in food additives It may be included, and may further include suitable carriers, excipients and diluents commonly used in the manufacture of food acceptable food supplement additives or food additives.

For example, the functional food additive composition, which is effective in preventing liver damage and preventing liver damage, including a salt having improved liver protection function and antioxidant activity as an active ingredient, prepared by the manufacturing method, includes various nutrients, vitamins, and minerals. (Electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers , Preservatives, glycerin and the like. These components can be used independently or in combination. The ratio of such additives is not so important, but may be selected in the range of 0 to 20,000 parts by weight per 100 parts by weight of salt, which is improved in the hepatoprotective function produced by the production method of the present invention and the antioxidant capacity is improved.

In addition, the intake status and weight of the functional food additive composition, which is effective in protecting the liver and preventing liver damage, including the salt having an improved anti-oxidant activity as an active ingredient, which has been improved by the manufacturing method, and has an antioxidant activity. Depending on the drug form, route of administration, and duration, it may be appropriately selected by those skilled in the art. Preferably, the food additive composition of the present invention is 0.0001 to 12 g / kg per day (based on the amount / weight of functional salts) based on the improved salt protection function and the antioxidant activity is improved by the production method as an active ingredient May be administered at 0.01 to 9 g / kg to be more effective. The method of administration may be administered once a day, may be administered several times, and the dosage does not limit the scope of the present invention in any aspect.

The salt produced by the preparation method of the present invention is improved and the antioxidant activity is improved by using DPPH and iron ion reducing power. As a result, it was confirmed that the antioxidant activity was remarkably excellent. This has been found to be excellent and not cytotoxic, and has been found to be widely used in connection with liver protection and prevention of liver damage.

As described above, according to the present invention, the manufacturing process is simple and can significantly improve the antioxidant capacity of the salt in an economical way compared to the existing method, since the salt prepared by the method is very excellent in the antioxidant capacity, Functional food additive composition which is effective in eliminating free radicals in the body to prevent or improve liver damage caused by oxidative stress, as well as protecting liver and preventing liver damage using the salt prepared by the above method as an active ingredient. Not only does it protect the liver and prevent liver damage, but it is a natural substance that is used as an active ingredient unlike conventional compounds used for protecting the liver or preventing liver damage. In particular, natural salt produced in Jeollanam-do can be used as a raw material. Typically it would also be advantageous in that a very large effect industrially.

Hereinafter, preferred embodiments of the present invention will be described. However, the following examples are only preferred embodiments of the present invention, and the present invention is not limited to the following examples.

Example  One: Antioxidant activity  Preparation of Improved Functional Salt

Solar salts used in the experiments were purchased from Taepyung Salt (Korea), refined salts were purchased from Hangeum (Korea), and NaCl (PS, Purified salts) was S9625 (Sigma, USA, NaCl 99.5%). Was purchased and used, purple bamboo salt (PBS) and key power salt (BS, Bamboo salt) was purchased from fresh water (Korea).

Bamboo was purchased from plastic & bamboo, and MgSO ₄ and bamboo were purchased from Farm Maker. Functional salts with improved antioxidant capacity are processed salts prepared using reagent grade salts according to the raw materials, Processed purified salts (P-PS) and Processed solar salts (P-SS) for functional salts prepared using sun salt. ) And functional salts prepared using baked salt through one heat treatment, respectively, were named P-RS (Processed Roated salts).

Preparation of the functional salt corresponding to the present invention is carried out by a method of heat-treating the natural salt divided into two times. The heat treatment corresponding to the pre-treatment of the sun salt was performed by heat-treating the sun salt at 800 ° C. for 2 hours. Heat treatment corresponding to the main treatment of the sun salt was performed by applying heat treatment under the conditions of FIG. 1 and Table 1 below.

Addition of subsidiary materials to confirm the optimal condition of functional salts with antioxidant and hepatoprotective functions was performed in 65 parts by weight based on 1 part by weight of functional salts (salt: MgSO ₄ : Mixed), and in the case of functional salt 2, 60: 40 (salt: bamboo) based on the weight part, and in the case of functional salt 3, 60: 30: 10 (salt: bamboo: MgSO ₄ ).

TABLE 1

Step Temp (℃) Step duration
in h: min Temp-min
(℃ / min) Temp gradient type One 200 01:40 2 Gradient 2 400 00:50 4 Gradient 3 1,100 02:55 4 Gradient 4 1,100 00:30 - Step-n-hold

TABLE 2

More specifically, each mixture of the raw material salt and the additive was added to a ceramic container made of alumina was prepared by heating in an electric furnace (electric heating, Korea) for the time described in FIG.

Example  2: Characterization and Characterization of Functional Salt

First of all, NaCl was measured by Mohr method, and about 1 g of sample was diluted with distilled water, and then diluted to 100 mL, followed by 10 mL of 10% potassium chromate (K ₂ CrO ₄ ) indicator. It was titrated with nitric acid solution (AgNO ₃ ), and calculated according to the following formula 1 and expressed as the amount of salt.

[Calculation 1]

In addition, the pH was measured using a pH meter (Model 730p, Istda, Korea) in 1% salt solution, and the oxidation-reduction potential (ORP) was measured, respectively, and the results are shown in Table 3. .

In addition, the measurement of the mineral component is dissolved in each of the functional salt in 0.1% nitric acid solution, filtered by pure water without salt, and diluted to a certain concentration in consideration of the concentration of each element to be analyzed, to prepare an experimental solution. Then, using the prepared experimental solution, the mineral component and its content were analyzed using an atomic absorber (Atomic absorption spectrophotometer, Spectra AA 220Fs, Varian Co., Australia). In the analysis using the atomic absorber, 20 ml of a mixture of nitric acid and perchloric acid mixed at a volume ratio of 4: 1 was added to a sample containing the functional salt, and thermally decomposed at 100 ° C., and neutralized with ammonia, and then nitric acid solution. The concentration of each component in the atomic absorber was measured by diluting to a concentration corresponding to the element to be analyzed using the results, and the results are shown in Table 4.

[Table 3]

Sample name ¹⁾ DDW PBS pH
ORP
(mV) pH
ORP
(mV) P-RS (1) 9.17 15 6.83 78 P-RS (2) 10.66 -239 7.22 -243 P-RS (3) 9.66 -48 6.88 42 P-SS (1) 9.32 135 6.84 72 P-SS (2) 10.21 -225 7.04 -206 P-SS (3) 9.82 -79 7.11 -20 P-PS (1) 8.36 170 6.84 83 P-PS (2) 10.52 -236 7.29 -249 P-PS (3) 9.92 -37 7.06 20 P-RS 8.77 78 7.0 60 P-SS 9.06 102 6.94 87 P-PS 5.75 173 7.06 95 RS 10.55 20 7.58 42 SS 8.05 88 7.09 92 PS 6.1 120 7.09 106 BS 11.12 -128 7.76 -128 PBS 10.66 -224 7.24 -243

[Table 4]

 Sample Major elements (ppm) minor elements (ppm) NaCl
(%) Na K Cl
(Cl ^-
ion) Ca Mg S
(SO ₄ ^{2 -}
ion) Fe Zn Cu Mn Sr Br
(ion) PPS2 373,000 1,265 574,690 95 14,971 6,716 17.79 1.54 1.88 3.62 0.30 368 88.3 PSS2 387,000 5,576 605,557 1,643 12,211 1,714 20.77 1.95 1.83 7.41 7.16 704 94.9 PRS2 387,750 4,606 599,550 1,942 12,146 2,628 33.42 2.02 1.67 6.63 7.58 546 93.6

As shown in Table 3, irrespective of the type of salt corresponding to the present invention, in the case of functional salt 2 to which only bamboo was added as a subsidiary material, a negative value was measured based on ORP (redox potential) to confirm that reducing power was formed. In particular, in the case of sun salt (PRS-2) corresponding to the present invention, it was confirmed that the most excellent antioxidant capacity improvement effect on the basis of reducing power formation. The results described in Table 3 above are shown in Table 5 below.

In addition, as shown in Table 4, when using the natural salt, it was confirmed that the mineral content is remarkably improved compared to the case of using the reagent refining salt, the recent consumer who requires the mineral supply when the salt intake associated with the intake of salt Considering the tendency of the improvement of the mineral content was evaluated that the natural salt, in particular, the natural salt after heat treatment once has the best effect.

TABLE 5

Sample Name Reagent grade
Salt (PS) bamboo
adding MgSO ₄ Death standardization
Burning process pH
(in DDW) ORP
(in PBS) compare RS ○ - - - - 10.55 7.58 Untreated control PRS ○ - - - ○ 8.77 7.0 No Subsidies PRS-1 ○ - ○ ○ ○ 9.17 78 Subsidiary fee addition group PRS-2 ○ ○ - - ○ 10.66 -243 Subsidiary fee addition group PRS-3 ○ ○ ○ - ○ 9.66 42 Ingredient Addition Group

Example  3: of functional salt Antioxidant activity  Measure

3-1. DPPH Using Antioxidant activity  Measure

The antioxidant capacity of the functional salt and the raw material salt prepared in Example 1 was measured using DPPH radical scavenging ability, and the measurement method was measured by modifying the method of Abe and Yamacuchi.

First, 400 μL of 100 mM Tris-HCl buffer (pH 7.4) was added to 100 μL of the salt or functional salt sample, followed by 200 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH, Sigma, ST. Louis, MO, USA). 500 μL of Methanol solution was added. The mixture was mixed with a Vortex mixer and reacted in the dark for 10 minutes, and the absorbance was measured at 517 nm. Scavenging activity was measured by the absorbance of DPPH radical itself to the DPPH radical scavenging absorbance of the sample, the results are shown in Figure 2 and 3 and Table 6.

[Equation 2]

TABLE 6

Samples DPPH radical scavenging activity
(IC ₅₀ ) PS ND SS ND RS ND BS 〈100 PBS 8.75 PPS2 4.24 PSS2 22.15 PRS2 3.49

As shown in FIG. 2 and FIG. 3, in the case of the functional salt 2 treated only with bamboo of the present invention, the remarkable DPPH radical scavenging ability was confirmed, compared with the salt treated by the other method, and it was confirmed that it was excellent in antioxidant activity. In addition, in the case of the baked salt heat-treated one time the natural salt of the present invention, it was confirmed that there is a remarkably excellent antioxidant capacity compared to the case of using a simple natural salt or using a purified salt, about 450 compared to the porridge salt that is reported to have excellent antioxidant capacity in general It was found to have good antioxidant capacity of%. This difference in antioxidant capacity can be more clearly confirmed by comparing the IC50 values described in Table 6 above. According to Table 8, it was confirmed that in the case of the baked salt obtained by heat-treating the natural salt of the present invention once, only about 30% of the bamboo salt could be used to obtain the same result as the bamboo salt.

In the case of the salt of the present invention, by performing the heat treatment process twice, the treatment process is simple and the cost is reduced compared to the porridge salt which performs the heat treatment process a total of nine times, the antioxidant capacity is comparable or remarkably excellent, As an alternative antioxidant salt, the industrial effect was expected to be very large.

In addition, in the case of the antioxidant capacity according to the concentration of the sample salt, as shown in Figure 3, it was confirmed that the antioxidant capacity is improved in proportion to the salt treatment concentration.

3-2. Antioxidant Activity Measurement Using Iron Ion Reducing Power

The antioxidant power measurement using iron (Fe ³⁺ ) ion reducing power is a method of measuring the reducing power of converting ferric (Fe ³⁺ ) state into ferrous state (Fe ²⁺ ) in the sample solution using the method of Benzie et al. (1996). Was performed. In the state of Fe ³⁺ -TPTZ complex, electrons are donated by antioxidant or reductant to be reduced to Fe ²⁺ -TPTZ comples, and the color becomes blue.

Specifically, 100ul of ferrous reducing antioxidative power (FRAP) prepared by mixing 10mM 2,4,6-tripyridyl triazine (TPTZ), 0.25M acetate buffer (pH 3.6) and 20mM ferric chloride in a ratio of 1: 10: 1, respectively. 30ul of each salt sample and 90ul of distilled water were added and reacted for 4 minutes. The control group was reacted with distilled water instead of the sample. The results were expressed as the concentration (mM) of the sample capable of reducing 1 mM ferrous sulphate, and the results are shown in FIG. 4.

As shown in FIG. 4, the functional salt produced by the method of the present invention has an improved antioxidant capacity, the existing reducing power is reported to have an antioxidant capacity to be equal to or superior to the bamboo salt or porridge salt In particular, in the case of the baked salt heat-treated once a natural salt, it was confirmed that it has a remarkably excellent reducing power compared to the porridge salt.

Example 4 Measurement of Hepatoprotective Function

4-1. Cytotoxicity Measurement

Cytotoxicity was used for HepG2, a human liver cancer cell line passaged and cultured from the Bank of Korea Cell Line. The medium for cell culture was MEM-α containing penicillin G (100 IU / mL) and streptomycin (100 mg / mL). FBS was mixed to 10% of the total amount and used to adjust to pH 7.4.

The cells were cultured in a CO ₂ incubator adjusted to 37 ° C. and 5% CO ₂ , and passaged in 75-cm ² plastic flasks (Nunc, Denmark) once every 2-3 days to maintain cell lines.

Hepatotoxicity measurement of the present invention was performed by the following method. Specifically, HepG2 cells, the cultured human liver cancer cell line, were treated with 0.05% Trypsin-EDTA to isolate cells, and adjusted to 1 × 10 ⁵ cells / mL for each well, placed in 100μL wells, and then at 37 ° C. humidified 5% CO ₂ And 95% air incubator (Sanyo, USA) for 24 hours pre-culture. To confirm the inhibition of HepG2 cell growth, the cultured cells were seeded in 96 well plates and incubated 24 hours before at 37 ° C., 5%, CO ₂ incubator. Functional salt of the present invention and the existing salt was treated with 10,000ug / ml and incubated in the same conditions as the pre-culture. 100 μl of MTT solution (Sigma, USA) having a final concentration of 0.4 mg / ml was added to the culture solution, and the reaction solution was removed for 4 hours and the supernatant was removed. After 200 µl of dimethylsulfoxide (DMSO: Sigma, USA) was added to dissolve the precipitate, the absorbance was measured at 570 nm by MISA to form blue formazan by mitochondrial dehydrogenase in mitochondria of living cells. 5 is shown.

As shown in FIG. 5, in general, the salt was found to have very low cytotoxicity even when treated with 10,000 ug / ml. In particular, in the case of the functional salt produced by the method of the present invention, the cytotoxicity was improved. It was confirmed that there was little, and excellent cell safety was confirmed compared with other salts. Therefore, it is evaluated that the salt of the present invention can be used for various purposes including food or medicine.

4-2. Measure liver protection

Liver protection was measured using a 12-week-old Sprague-Dawley rat (Damul Science Co., Ltd., Korea). The breeding of the test animals was adapted to the diet of the Harlan Rodent diet (2018S) and weighed by measuring their weight. This was done by classifying into experimental group of. The breeding conditions of the experimental animals were carried out at 20 ℃ and humidity conditions of 50 to 60% and day and night lighting conditions of 12 hours, water and diet were to be ingested freely.

The feed used in this study was a high-salt feed formulated with Rodent Purified diet (2018S, Harlan Co., USA) with each salt adjusted to 8% based on NaCl content. CCl ₄ used in the experiment was prepared by diluting 1: 1 using olive oil and intraperitoneally injected at a content of 2 mg / kg (CCl ₄ / body weight). The positive control group used in the experiment was treated with olive oil at a content of 2 mg / ml, the negative control group was treated with CCl ₄ and olive oil mixture at a content of 2 mg / kg, and each experimental treatment group ingested each salt. , CCl ₄ and the olive oil mixture is treated with a content of 2mg / kg.

In the plasma GOT measurement, blood collected from the test animal was left at room temperature for 15 minutes, and then centrifuged at 3,000 rpm for 10 minutes to separate plasma, and an automatic blood analyzer (VITRIOS 60 automatic Blood Chemistry analyser, Johnon & Johnon , USA) to measure GOT and GPT values. GOT and GPT of all the experimental animals were measured during the test period at intervals of 0 hours from the beginning of the experiment, the results are shown in FIG. Figure 6 is the average of the measured values of three experimental animals for each experimental group.

GOT is an abbreviation of enzyme called glutatmic oxaloacetic transaminase. It is an enzyme present in hepatocytes. When the cell membrane is destroyed by liver damage, it is leaked out of the cell and the value is increased. Damage to hepatocytes increases the amount of these enzymes released into the medium, and drugs with hepatoprotective action have been reported to reduce the amount of these enzymes released.

As shown in FIG. 6, the functional salt produced by the manufacturing method of the present invention was measured to significantly reduce the GOT content compared to the roasted salt after heat treatment of the natural salt once. In addition, it was determined that the GOT content is slightly reduced compared to the sun salt, and was found to be the best method for improving the hepatoprotective function of the sun salt.

1 is a graph showing a heat treatment process of a method for improving the antioxidant capacity of the sun salt according to an embodiment of the present invention.

2 is a graph measuring the antioxidant capacity of the functional salt measured using DPPH according to an embodiment of the present invention.

Figure 3 is a graph measuring the antioxidant capacity of each functional salt concentration measured using DPPH according to an embodiment of the present invention.

Figure 4 is a graph showing the reducing power of the functional salt measured by using the iron reducing power according to an embodiment of the present invention.

Figure 5 is a graph showing the cytotoxicity of functional salts measured cytotoxicity measured using MTT Assay according to an embodiment of the present invention.

6 is a graph showing the hepatoprotective action of functional salts measured using GOT values according to an embodiment of the present invention.

Claims (5)

a) a first heat treatment step of heat-treating the natural salt for 1 hour to 12 hours at 700 to 900 ℃ b) mixing the natural salt and bamboo which have been subjected to the first heat treatment to 40:60 to 70:30 (natural salt: bamboo) based on a weight ratio; c) a second heat treatment step of heating the sun salt and bamboo mixture for 4 to 6 hours and heating the mixture until 1,100 ° C .; And d) a third heat treatment step of heat-treating the mixture on which the second heat treatment step is performed at 1,100 ° C. for 15 to 60 minutes Hepatoprotective function is improved, including the method of producing a salt with improved antioxidant capacity. According to claim 1, wherein the step c) is applied to heat the mixture so that the temperature of the mixture rises 2 ℃ per minute when the temperature of the mixture ranges from 0 ℃ to 200 ℃, the temperature of the mixture is from 200 ℃ to 400 ℃ In the range is applied to heat the mixture so that the temperature rises 4 ℃ per minute, and when the temperature of the mixture in the range of 400 ℃ to 1,100 ℃ by heating to increase the temperature 4 minutes per minute to improve the hepatoprotective function is improved And a method for producing salt having improved antioxidant capacity. delete delete delete

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