KR20040075643A - Method for production of a sweet drink made from fermented rice with anti-cancerous effect by adding herb remedies and a sweet drink thereof - Google Patents

Abstract

PURPOSE: A method of making sweet rice drink(shikye)containing powders of herbal plants such as Ulmus davidiana, Gardeniae Fructus, Puerariae Radix, mulberry leaf, Glycyrrhizae Radix and ginger is provided. The product has the functionality of preventing cancer as well as characteristic taste, flavor and color of shikye. CONSTITUTION: A mixture of 25 parts by weight of glutinous rice and 25 parts by weight of coix seeds is immersed in 100 parts by weight of water for 2hr at room temperature and steamed for 10min at 120deg.C to give shikye cooked rice. A mixture of 47 parts by weight of malt powder(yotkirum), 0.5 parts by weight of Ulmus davidiana powder, 0.5 parts by weight of Gardeniae Fructus powder, 0.5 parts by weight of Puerariae Radix, 0.5 parts by weight of mulberry leaf powder, 0.5 parts by weight of Glycyrrhizae Radix powder and 0.5 parts by weight of ginger powder is added to 500 parts by weight of purified water and extracted for 2hr at 60deg.C to give malt extract. Thereafter, the shikye cooked rice is mixed with the malt extract and saccharified for 5hr at 50deg.C.

Description

Method for production of a functional herbal Sikhye and cancer prevention {Method for production of a sweet drink made from fermented rice with anti-cancerous effect by adding herb remedies and a sweet drink about}

The present invention relates to Sikhye, and more specifically, the malt liquid obtained by hot water extraction after adding elm bark powder, gardenia powder, brown root powder, mulberry leaf powder, licorice powder and ginger powder to malt powder is obtained from glutinous rice and yulmu. It is related to herbal Sikhye prepared by saccharification by addition to Sikhye rice.

Sikhye is a traditional Korean drink that has been greatly loved by families since the Joseon Dynasty.

In the general manufacturing method of Sikhye, first, sift the fine malt powder sifted into a bag, extract it from the water at about 50 ° C., and then sediment is settled. The malt is prepared by pouring the prepared malt water into rice cooked with non-glutinous rice or glutinous rice and keeping it at 50 ° C. for 4-5 hours. The rice grains are glycosylated by the enzymatic action contained in the malt.

If a few grains come out, scoop them out, rinse them in cold water, and then drain them. Then, add sugar to the brewed foods, boil them, and cool them to make Sikhye.

In addition to the increase in palatability for Sikhye, research is being actively conducted to increase palatability of Sikhye as it is commercialized and mass produced and sold.

On the other hand, as a conventional technique,

By adding hemp to Korean Patent Publication No. 10-0296729, the taste and aroma are improved, and Sikhye is disclosed to maintain and promote the health of the human body when drinking.

In the Republic of Korea Patent Publication No. 1998-068217 is prepared by mixing Ganoderma lucidum, Sol 닢, ginseng, 칡, licorice and the like, the manufacturing method of Sikhye is unusual and has a simplified manufacturing process is disclosed,

By manufacturing ginseng or Schisandra chinensis in Republic of Korea Patent Publication No. 1997-0058599 discloses a method of producing Sikhye added the health functional and unique flavor of herbal ingredients,

Sikhye has been disclosed to have the effect of weight loss and prevention of hypertension by adding herbal medicines such as tofu leaf, green tea, buckwheat leaf, persimmon leaf, mountain lion, Dong Kyuja to Republic of Korea Patent Publication No. 10-0173840.

However, there has been no prior art related to Korean dietary supplement with cancer prevention effects.

Therefore, the present inventors have repeatedly studied to provide functional health Sikhye effective in cancer prevention with the palatability of Sikhye, and as a result, elm powder, gardenia powder, root powder, mulberry leaf powder, licorice powder and ginger powder in malt powder After mixing the malt solution obtained by hot water extraction to glutinous rice, Sikhye rice prepared from Yulmu and saccharifying, by leaching into the active ingredient Sikhye of the herbal medicine, it was found that Sikhye reinforced with the functionality of cancer prevention can be prepared. The present invention has been completed.

Accordingly, an object of the present invention is to provide a herbal Sikhye having a cancer prevention effect and a manufacturing method by adding the malt solution from which the herbal medicine is leached to the Sikhye rice prepared from glutinous rice, Yulmu and saccharified.

In order to achieve the above object, the present invention,

Mixing 25 parts by weight of glutinous rice and 25 parts by weight of yulmu, pouring 100 parts by weight of water and soaking at room temperature for 2 hours, and then cooking for 10 minutes at a temperature of 120 ° C. to prepare Sikhye rice;

47 parts by weight of malt oil, 0.5 parts by weight of elm bark powder, 0.5 parts by weight of gardenia powder, 0.5 parts by weight of root powder, 0.5 parts by weight of mulberry leaf powder, 0.5 parts by weight of licorice powder and 0.5 parts by weight of ginger powder Hot water extraction at a temperature of 60 ° C. for 2 hours to obtain malt solution; And

After mixing the prepared Sikhye rice and malt solution prepared by the above step provides a method for producing herbal Sikhye comprising the step of saccharifying for 5 hours at 50 ℃.

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

Herbal medicines added to enhance the functionality of Sikhye in the present invention are elm bark, gardenia, roots, mulberry leaves, licorice and ginger, the elm bark is used for moisturizing, diuretic, small bovine poison (독) , Gardenia is used for the treatment of insomnia and jaundice. It has the effect of anti-inflammatory, hemostasis and diuresis. The roots are sweating, fever, and laxative in Chinese medicine, high fever, headache, hypertension, heart failure, diarrhea, and shoulder Used when Mulberry leaves have the effect of heat lowering, coughing, licorice has the effect of improving sweetness, ginger is effective in indigestion, vomiting, diarrhea, promotes blood circulation, anti-inflammatory and analgesic effect.

In the present invention, the malt solution to which the herbal agent is added is prepared by adding the herbal medicine during malt solution preparation.

During the production of Sikhye, during the malt production or saccharification process, it is possible to extract the herbal medicines in parallel with the malt production process and the saccharification process by adding the herbal medicine to the malt production process. The addition of the herbal medicine to the manufacturing process has the advantage of less bitterness, thus adding a large amount of herbal ingredients.

This is because, in the present invention, the temperature is higher during the production of the malt solution, the effect of hot water extraction is excellent, and at the same time it is possible to more efficiently leach the active ingredient from the herbal medicine by the action of various enzymes derived from malt at the same time leaching.

In addition, the present invention can extract the active ingredient from the herbal medicine without a separate extraction process, there is an advantage to simplify the process.

Herbal medicines used in the health Sikhye of the present invention is not particularly limited to the form, for example, the water content is dried to 10% or less so that the leaching of the active ingredient is easily made, and then pulverized to 0.1 ~ 0.5cm use.

In addition, the herbal medicines used in the present invention have a strong intrinsic taste and a characteristic irritating odor, which may impair the overall palatability of Sikhye, so that 3.0 wt% of malt oil is added equally to the weight of water. On the other hand, the prepared herbal foods can be ingested by adding an additional sweetener according to preference.

Sikhye of the present invention prepared as described above has the effect of inhibiting cancer occurrence as a result of the experiment by the following experimental example.

Hereinafter, the configuration of the present invention will be described in more detail with reference to the following examples, but the scope of the present invention is not limited only to these examples.

Example 1: Preparation of herbal Sikhye of the present invention

After mixing 25 g of glutinous rice and 25 g of yulmu, 100 g of water was poured and soaked at room temperature for 2 hours, and then cooked for 10 minutes at a temperature of 120 ° C. to prepare Sikhye rice.

In addition, to 500 g of purified water, 47 g of malt oil, 0.5 g of elm bark powder, 0.5 g of gardenia powder, 0.5 g of root powder, 0.5 g of mulberry leaf powder, 0.5 g of licorice powder, 0.5 g of licorice powder and 0.5 g of ginger powder were added, and then the temperature was 60 ° C. Hot water was extracted for 2 hours to obtain malt solution.

After mixing the Sikhye rice and malt solution prepared in the above step 2 and saccharified for 5 hours at 50 ℃ to prepare a herbal Sikhye.

Comparative Example 1: Preparation of general Sikhye prepared by the same process as the present invention except adding the herbal medicine

In the production of malt, general Sikhye was prepared in the same manner as in Example 1 except for adding a herbal medicine. However, the amount of malt added at this time was 50 g.

Experimental Example 1: sensory test

The taste of Sikhye was scored based on the following evaluation criteria for Sikhye of Example 1 and Comparative Example 1, targeting 10 professional sensory test personnel. The evaluation was repeated three times and the values are shown in Table 1.

Panel 1 Panel 2 Panel 3 Panel 4 Panel 5 Panel 6 Panel 7 Panel 8 Panel 9 Panel 10 total score 4 5 3 4 5 3 3 5 4 4 40 The score indicates the superiority and inferiority of the Korean dietary supplement prepared in Example 1 compared to the general Sikhye prepared in Comparative Example 1, "very good" 5 points, "excellent" 4 points, "similar". 3 points, "bad" 2 points, "very bad." 1 point was given.

From the results of Table 1, the herbal Sikhye of the present invention, unlike the conventional prediction, despite the addition of herbal medicines, has almost the same flavor as general Sikhye, and I tend to be aware of the health food from the unique flavor of Chinese medicine By doing this, it turned out that the rather favorable flavor is shown.

Experimental Example 2: Effect of the Herbal Medicine Sikhye on Inhibiting Solid Cancer Growth

In the present experimental example, Balb / c mouse to see the anti-tumor effect by Sikhye obtained from Example 1 (hereinafter referred to as "Hansikhye") and Sikhye obtained from Comparative Example 1 (hereinafter referred to as "General Sikhye") After implanting 'sarcoma-180' tumor cells in the left inguinal subcutaneous of the 0.25 mg / kg sample was administered for 20 days, after 32 days from the mouse was dissected tumor was isolated and weighed.

As a result, as shown in Table 2, the control group implanted with only tumor cells (S-180 + PBS, administered with the same amount of PBS instead of the sample) had a tumor weight of 5.98g, whereas the group injected with Hansikhye (S-180 + FS) decreased to 4.45g, showing significant tumor suppression effect, and the general Sikhye injected group (S-180 + CS) was 5.90g, which was not significantly different from the control group.

Therefore, it was found that Han, Hye-hye had a solid growth inhibition effect.

Tumor weight measurement after Sikhye administration to Balb / c mice transplanted with 'sarcoma-180' cells Sample Dose (mg / kg) Tumor Weight (g) S-180 + PBS 0.25 5.98 ± 0.02 S-180 + CS ¹ 0.25 5.90 ± 0.74 S-180 + FS ² 0.25 4.45 ± 1.12 [Note] 1: General Sikhye 2: Han Hye-hye

Experimental Example 3 Weight Changes of Organs in Mice fed Sikhye

In order to measure the effects of the present invention, Hansikhye and general Sikhye in other organs other than the tumor, the control (Control) was only free diet, the tumor cell control group (S-180 + PBS) free diet and injection PBS The treated group was given a free diet and injected with the same amount of tumor cells (Sarcoma-180 cells) after transplanting the sikhye with the same amount of tumor cell control group, the sample was administered for 20 days and the weight of each organ Measured.

As a result, as shown in Table 3, the weight ratio of liver, which is responsible for the detoxification of various harmful substances generated during metabolic processes, was the control group implanted with tumor cells only (S-180 + PBS) and the group injected with general Sikhye (S- 180 + CS) was slightly higher than S-180 + FS, and spleen was lower than the control group implanted with tumor cells. There was no big difference.

Weight of each organ of the mouse fed dietary medicinal herb and general sikhye of the present invention Sample Weight (g) Spleen / Weight (%) Liver / Weight (%) Heart / Weight (%) Height / weight (%) Control 28.85 ± 1.8 0.35 ± 0.01 5.71 ± 0.5 0.51 ± 0.14 1.87 ± 0.06 S-180 + PBS 29.4 ± 1.4 1.57 ± 0.10 7.73 ± 0.8 0.33 ± 0.02 1.59 ± 0.03 S-180 + CS ¹ 29.8 ± 1.3 1.45 ± 0.10 7.56 ± 0.7 0.32 ± 0.03 1.68 ± 0.19 S-180 + FS ² 28.5 ± 2.0 1.39 ± 0.02 6.69 ± 0.3 0.36 ± 0.02 1.65 ± 0.05 [Note] 1: General Sikhye 2: Han Hye-hye

Experimental Example 4: Effect of Increased Immune Activity of the Invention Herbal Medicine and General Sikhye

In this experimental example, the tumor cells were transplanted in the same manner as in Experiment 3, and the spleens of each mouse were sterilely extracted after 20 days of administration of the present invention, the method of the present invention, and the natural killer (NK) of the spleen. ) Cell activity was examined.

In order to analyze the activity of natural killer cells, first, effector cells (NK cells) were prepared. The spleens of the mice were aseptically extracted in a clean bench, washed three times with 5 mL of RPMI 1640 medium containing penicillin (100 U / mL) and streptomycin (100 μg / mL), and then finely ground.

The cell suspension was filtered through a 70 μm nylon mesh, centrifuged to collect lymphocytes, and then suspended in the same medium.

Lymphocytes were isolated by centrifugation (500 × g, 30 min, 18 ° C.) using histopacque-1077 (histopaque-1077). The prepared cells were resuspended in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamine, penicillin (100 U / mL), streptomycin (100 μg / mL).

This was incubated for 1 to 2 hours at 37 ° C. in a CO ₂ incubator to allow cells to adhere to the culture flask (6 cm Petri dish). Non-adhesive natural killer cells were collected by centrifugation, resuspended in culture medium, and the appropriate number of cells was counted. Subsequently, cell destruction activity of natural killer cells was analyzed by MTT method.

Yac-1 mouse lymphoma cells were used as target cells. Yac-1 cells are the most convenient target cell line for measuring NK cell activity.

Yac-1 cells were diluted to a density of 5 × 10 ⁴ cells / mL in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamine penicillin (100 U / mL) and streptomycin (100 μg / mL). 50 μl was added to the wells.

In addition, 50 μl of effector cells and Yac-1 cells were added to 96-well plates for NK cell activity analysis of NK cells × 10 ⁷ cells / mL.

After incubating for 3 days at 37 ° C. in a CO ₂ incubator, 10 μl of MTT (5 mg / mL) solution was added to each well and incubated at 37 ° C. for 4 hours. 25 μl of SDS (10% in 0.02N HCl) was added thereto, followed by color development at room temperature for 30 minutes, and OD was measured at 540 nm. Percent cytotoxicity was calculated by the following formula.

The results showed that the S-180 + FS group showed the highest inhibitory effect of 55.42%, 49.35% in the control group, 31.52% in the transplantation of tumor cells only, and transplanted with the general Sikhye group. Was 33.42%. In the above results, the group receiving general Sikhye showed low NK cell immune activity, similar to the group transplanted with only tumor cells.

Experimental Example 5: Glutathione-S-transferase Activity Changes in Mouse Liver by Invention of Korean Invention

The phase II phase of the metabolic enzyme system in the liver removes foreign substances by including endogenous or externally administered toxic substances or by converting them into water-soluble substances to release them to the body. Glutathione-S-transferase (glutathione- S-transferase) is an enzyme that is involved in detoxification by transferring and excreting toxic substances and peroxides in the body using reduced glutathione.

In this experimental example, the glutathione-S-transferase activity in the rats fed the sample and the diet was measured as in Example 4.

First, in order to measure glutathione-S-transferase (GST) activity, mice were killed and the liver was perfused with physiological saline of 4 ° C. or lower to remove blood remaining in the liver, and the liver was extracted.

0.1 M potassium phosphate buffer (pH 7.4) was added per 1 g of liver tissue, and the mixture was ground with a glass teflon omogenizer under ice cooling.

The ground liquor was used as a homogenate fraction, which was centrifuged at 13,000 rpm for 10 minutes to remove nuclei and unbroken cell sections, and the supernatant obtained by ultracentrifugation at 105,000 × g for 1 hour was used as a cytosol fraction. It was made.

75 μl of 0.04 M reduced glutathione was added to 0.1 M potassium phosphate buffer (pH 6.5), 0.1 mL of enzyme solution was added, and 0.5 mL of 20% trichloroacetic acid was added to the blank to inactivate the enzyme, and the sample was reacted at 25 ° C. for 5 minutes. 25 μl of 0.12M 1-chloro-2,4-dinitrobenzene was added to each sample and reacted at 25 ° C. for 2 minutes. 20% trichloroacetic acid was added to the sample to complete the reaction, followed by centrifugation. After absorbance was measured at 340 nm, the activity was calculated using a molar extinction coefficient of 1-chloro-2,4-dinitrobenzene 9.6 mM ¹ cm ^-1 .

Units of enzymatic activity were expressed as nmole number of 2,4-dinitrobenzene-glutathione produced by mg protein for 1 minute.

As a result, glutathione-S-transferase activity was 391.5nmol / mg protein / min in the control group without transplanting tumor cells and 282.5nmol / mg protein / min in the control group in which tumor cells were transplanted (S-180 + PBS). fell.

In the group receiving general Sikhye (S-180 + CS), it was lower than the control group implanted with tumor cells at 275.4 nmol / mg protein / min, but in the group receiving Han Sikhye (S-180 + FS) The activity of glutathione-S-transferase was significantly higher than that of the control group transplanted with 512.5 nmol / mg protein / min.

Experimental Example 6: Glycogen Distribution in Mouse Liver Tissues of the Invention

In this example, the glycogen distribution in the liver tissue of rats fed the sample and the diet was examined as in Experimental Example 4. The rat liver was excised and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) at 4 ° C. for 12 hours, followed by sequential dehydration and clarification, and embedded in paraffin. A micrometer continuous section was produced.

Deparaffinization, hematoxylin-eosin (H-E) staining and periodic acid Schiff (PAS) reaction were performed for histological changes and glycogen distribution.

As a result, the tumor cells 'sarcoma-180' transplanted and PBS-administered group (S-180 + PBS) had liver-specific hepatic lobules, but had weak turbid swelling, bullous degeneration, fat degeneration and cupper (Kupffer). Hepatic cell changes, such as changes in cellular and local necrosis, are also observed.

In the control group, not only did the histological changes significantly increase, but also increased the number of eosinophilic cells and the cells of the nucleus-concentrating cells, especially in the periphery of the hepatic lobe.

Histologic changes were similar to that of S-180 + PBS in the case of dieting general Sikhye (S-180 + CS), and in the case of dieting Han Sikhye (S-180 + FS), fat degeneration and cupper cell changes were observed. However, there was a slight change in local necrosis.

This is summarized in Table 4.

Intrahepatic Tissue Changes in Mice Group / HC CS HD FC KC FN Normal + 0- + +-++ + 0- + S-180 + PBS +++ ++ +++ +++ ++ S-180 + CS +++ ++ +++ +++ ++ S-180 + FS +++ ++ ++ ++ + Note: HC: histological change, CS: boil formation, HD: hydrophilic alteration, FC: lipid change, KC: cupper cell response, FN: focal lesion 0: no change or negligible +: weakly ++: moderate Shown as a level +++: Shows bad ++++: Shows very bad

Glycogen distribution was decreased in the surrounding and intermediate regions of 'S-180 + PBS' compared to normal liver tissue, and 'S-180 + CS' and 'S-180 + FS' were also compared to normal liver tissue. It decreased slightly in the surrounding area. This indicates that the herbal dietary invention of the present invention causes less damage to hepatocytes than general sikhye. This is summarized in Table 5.

Glycogen Distribution in Mouse Liver group PZHL IZHL CZHL Normal ++++ ++-+++ ++-+++ S-180 + PBS +++ ++ ++-+++ S-180 + CS ++-+++ ++-+++ ++-+++ S-180 + FS +++ ++-+++ ++-+++ Note: PZHL: Peripheral section of hepatic lobe, IZHL: Intermediate section of mesenchymal lobe, CZHL: Central section of mesenchymal lobe, 0: No change or negligible +: Appear weak +++: Appear moderate ++++: Very bad

Experimental Example 7 Immunohistologic Responses of Mouse Liver Tissues by the Invention of the Invention

In this experimental example, in order to see the immunohistochemical response of tumor-associated proteins to liver tissues of rats fed with a sample and a diet as in Experiment 4, deparaffinizing 10 mM for immunohistochemical observations of proteins associated with each tumor Treatment was performed at 95 ° C. for 5 minutes in sodium citrate buffer (pH 6.0) and again placed in the same buffer at room temperature for 20 minutes.

This was treated with 3% methanol hydroperoxide at room temperature for 30 minutes and washed three times with PBS (0.01M, pH 7.5) three times for 5 minutes to remove the non-specific reactions, 'goat normal serum (Vector Lab., PK-6101). Treated at room temperature for 30 minutes.

Removed and Bax (sc-493), Bcl-2 (sc-783), Rb (sc-050), c-fos (sc-253), c-jun (sc-1694) from Santa Cruz Biotechnology Inc. ) Was diluted to 500: 1 and reacted in a 4 ° C wet room for 16 hours.

Washed with PBS and reacted 'biotinylated anti-rabbit lgG (Vector Lab., PK-6101)' for 30 minutes at room temperature, washed with PBS and ABC kit (avidin-biotin-peroxidae complex, vector Lab., PK- 6101) at room temperature for 60 minutes.

After washing with PBS again, it was developed for 5 minutes at room temperature with a 'DAB substract' kit (vector Lab., SK-4100). After washing with distilled water, control dyeing with 'Mayer's hematoxylin' was carried out to determine the degree of reaction under an optical microscope.

As a result, immunohistochemical reactions of tumor-associated proteins against hepatocytes showed only Bax, Rb, and c-fos among six antibodies, and showed stronger responses in the nucleus than cytoplasm, and also in inflammatory cells between some hepatocytes. Indicated.

Bax reacts only in the nucleus and decreases in 'S-180 + PBS' and 'S-180 + CS' compared to the normal group.

Rb increased in the nucleus and cytoplasm of S-180 + PBS and S-180 + CS compared to the normal group, especially in the cytoplasm of central venous hepatocytes. In the case of FS ', cytoplasmic change was small.

c-fos was increased in 'S-180 + PBS' compared to the nucleus and cytoplasm of normal group, but also increased similarly to control group in 'S-180 + CS' and 'S-180 + FS'. This is summarized in Table 6.

Immunohistochemical Responses of Tumor-associated Proteins in Rat Liver group Region / Antibody Bax Bcl-2 Rb c-jun c-fos Normal Nuclear cytoplasm ++++ 0 00 +++ 0- + 00 +++ 0- + S-180 + PBS Nuclear cytoplasm ++ 0 00 ++++++ 00 ++++++ S-180 + CS Nuclear cytoplasm ++-+++ 0 00 +++++ 00 ++++ S-180 + FS Nuclear cytoplasm +++ 0 00 ++++ 00 ++++-++ Note: 0: No change or negligible +: Appears weak +++: Appears moderate +++: Appears severe +++: Appears very bad

As described above, the present invention is a Sikhye rice prepared from glutinous rice, yulmu the malt solution obtained by hot water extraction after mixing the elm powder, gardenia powder, brown root powder, mulberry leaf powder, licorice powder and ginger powder to malt powder It is very useful in the health food industry because it has an excellent effect to produce Sikhye reinforced with the functionality of cancer prevention by adding to saccharification.

Claims (2)

Mixing 25 parts by weight of glutinous rice and 25 parts by weight of yulmu, pouring 100 parts by weight of water and soaking at room temperature for 2 hours, and then cooking for 10 minutes at a temperature of 120 ° C. to prepare Sikhye rice; 47 parts by weight of malt oil, 0.5 parts by weight of elm bark powder, 0.5 parts by weight of gardenia powder, 0.5 parts by weight of brown powder, 0.5 parts by weight of mulberry leaf powder, 0.5 parts by weight of licorice powder and 0.5 parts by weight of ginger powder Hot water extraction at a temperature of 60 ° C. for 2 hours to obtain malt solution; And The method of producing herbal Sikhye comprising the step of saccharifying for 5 hours at 50 ℃ after mixing the prepared Sikhye rice and malt solution prepared through the step. Herbal Sikhye having a cancer prevention function by being produced by the method of claim 1.