KR101038510B1 - prepartaion of concentrated syrups for oral use containing hydrolyzed products of Glycyrrhizae Radix extracts and its compounding herbal medicines - Google Patents

Abstract

The present invention relates to the preparation of an oral concentrated syrup containing a hydrolyzed extract of licorice and its complex herbal medicine, and more particularly, extract and hydrolysis by combining ginseng as a main agent, ginseng, bokyeong, raw sulfur, ginger and control. By converting the polysaccharides contained in herbal medicines into non-sugars, it enhances digestive tract absorption, enhances anti-inflammatory and analgesic effects, enhances antioxidant activity, strengthens stamina and immune function, and prevents deterioration and microbial contamination during storage. It is a method for producing an internally prepared concentrated syrup.

Licorice, hydrolyzed extract, absorption ability improvement

Description

Preparation of concentrated syrups for oral use containing hydrolyzed products of Glycyrrhizae Radix extracts and its compounding herbal medicines}

The present invention relates to a method for producing concentrated syrup for internal use containing a hydrolyzed extract of licorice and its complex herbal medicine. More particularly, fatigue inevitably suffers inevitably with social life under various environmental pollution and difficult economic conditions. Prevention and improvement of various circulatory diseases, digestive disorders, degenerative diseases, inflammation and pain caused by accumulation, obsession, mental tension, anxiety, emotional damage, digestive problems, sleep disorders and decreased immune function Among Chinese herbal medicines, licorice is the main ingredient, and ginseng, Bokryeong, raw sulfur, ginger, and control compound are manufactured as hydrolyzed extracts to promote gastrointestinal absorption ability, strengthen physical strength, improve various inflammation, protect liver, detoxification, cholesterol Licorice and its combined herbal medicines that can have a lowering effect, prevent and improve circulatory diseases It relates to a concentrated syrup production process of my taking containing decomposition extract.

Republic of Korea Patent Publication No. 10-2005-0108415

Republic of Korea Patent Publication No. 10-2006-0134057

In general, Ginger Gumcho-tang and Kyungokgo are used for reinforcing physical strength, improving various inflammations, protecting and detoxifying the liver, and preventing and improving circulatory diseases.

In particular, ginger persimmon tang has been used in herbal medicine, including ginseng, ginger and control, including licorice, and these ginger persimmon tang is lack of nutrition supplied to cardiomyocytes, myocardial contractility is not smooth, and bronchial and lung immune functions Degradation, lack of fluid in the oral cavity and bronchus, nourish the cardiomyocytes, oral cavity, bronchial mucous membranes and smooth the mucous membranes when the saliva is well on the lips, dry throat and mouth, cough and asthma and chest tightness. To boost immune function.

In addition, gyeongokgo has been treated as a folk remedy as a folk remedy for a long period of time, including ginseng, bokyeong, raw sulfur and honey are prescribed to strengthen stamina and stamina, enhance immune function, and show excellent efficacy in gastrointestinal and circulatory diseases.

However, in the case of extracting or aging with boiling water to produce the ginger persimmon tang or gyokgo, after combining constituent herbs such as ginseng, bokyeong, raw jihwang, ginger and control, including licorice, many active and auxiliary ingredients are complicated Reaction produced precipitates and aggregates, and the active ingredient was inactivated, which significantly lowered the pharmacological efficacy of the individual herb.

The present invention has been used for a long time in oriental medicine and folk remedies by showing the effect of strengthening stamina in the oriental medicine, promoting blood circulation, enhancing immune function, nourishing all cells, and activating brain cells to promote mental function Whey, nutrients to the myocardial cells to increase the contractile power of the myocardium, strengthen the function of the bronchial and lungs, promote blood circulation and promote the secretion of gastric juice secretion by combining the constituents of ginger persimmon tang to enhance digestion Improves unscientific manufacturing methods, improves the extraction efficiency of active ingredients, prevents alteration and microbial contamination during the storage and manufacturing of raw materials, thereby promoting blood circulation, improving digestive function, gastrointestinal protection, preventing bronchial damage, strengthening stamina and immune function To develop a manufacturing method that can maximize the efficacy, such as strengthening.

According to the method for producing concentrated syrup for internal use containing the hydrolyzed extract of licorice and its complex herbal according to the present invention, fatigue accumulation, mental tension, anxiety, emotional damage, digestive dysfunction, sleep disorders and immune deterioration Among the herbal medicines that are known to prevent and improve the inflammation and pain of various circulatory diseases, digestive diseases, and degenerative diseases, the licorice is the main ingredient, and ginseng, Fukryeong, Saenghwanghwang, ginger, and the control are combined into hydrolyzed extracts. By manufacturing, it increases the absorption ability of the active ingredient, promotes digestive function, strengthens stamina, improves immune function, protects and detoxifies liver, lowers cholesterol, prevents and improves circulatory diseases.

Ginger licorice tang to be applied to the present invention for achieving the above object has been used in herbal medicine prescribed by ginseng, ginger and control including licorice. Due to malnutrition supplied to cardiomyocytes, myocardial contractility is not smooth, bronchial and lung immune functions are deteriorated, oral and bronchial fluid is lacking. Asthma, when the chest is stuffy, nourish the cardiomyocytes, mouth, bronchus, pulmonary mucosa and smooth the mucous membranes to enhance immune function.

Gyungokgo has been treated as a folk remedy for a long time as ginseng, including Bokryeong, Saenghwang and Honey, which is prescribed as a folk remedy by reinforcing stamina and stamina, enhancing immune function, and showing excellent effects on gastrointestinal and circulatory diseases.

The present invention is prepared by the hydrolysis extraction and fermentation method of the constituent herbs of ginger persimmon tang and cheongogo respectively difficult and difficult to manufacture by converting the polysaccharide component contained in the herbal medicine to non-saccharides to enhance the absorption capacity of the active ingredient and deterioration And microbial contamination to the greatest extent possible.

In particular, when observing the efficacy of the active ingredient contained in the herbal medicine,

First, Glycyrrhizin (glycirrhizin) contained in licorice has anti-allergic effect, NK cell activation, interferon production, antipyretic effect, and anti-ulcer activity by inhibiting gastric acid secretion and protecting gastric mucosa.

Glycyrrhetic acid has anti-inflammatory, detoxification, antipyretic, gastrointestinal smooth muscle spasms and liver damage inhibitory effects by having steroid hormone-like action, and inhibits the activity of Phospholipase A2 (phospholipase A2). It inhibits Prostaglandin (prostaglandin) production and exhibits analgesic activity, as well as suppresses bronchial smooth muscle spasms, thereby showing antitussive and myocardial action.

LH-1 activates immune cells, enhances macrophage phagocytosis, promotes the production of immune memory cells, nourish cardiomyocytes, relieves cardiac contractions and normalizes cardiomotor activity.

Ligustrazine has anti-inflammatory activity, and β-Amyrin has antibacterial and antiviral effects, indicating antibacterial and anti-tumor activity against Staphylococcus aureus and Mycobacterium tuberculosis.

Second, Ginsenoside Rg1, Re, Rb2 (Ginseng Glycoside Rg1, Re, Rb2) contained in ginseng enhances adrenal cortex function and promotes immune cell production to enhance immune function, while Ginsenoside Rb1, Re and Adenosine (adenosine) It lowers the production of triglycerides in blood, and Ginsenoside Rg2 and Rg3 inhibit platelet coagulation and prevent thrombus formation.

Ginsenoside Rb1 has a platelet coagulation inhibitory effect, which prevents cerebral thrombosis, and has a lipolytic effect of fatty liver, which is used for the prevention and treatment of fatty liver due to alcohol drinking.It decreases Cholesterol (hypocholesterol) emia and activity of lymphocytes. Promotes immunity by promoting sedation and stabilizes nerves and improves insomnia.

Ginsenoside Rb2 inhibits cancer cell proliferation and Ginsenoside Rb2, Re, Rg1 enhances stamina by promoting protein synthesis, lipid synthesis and red blood cell production of bone marrow cells. Saponin has been reported to accelerate fatigue and accelerated condition reflexes to reduce fatigue and increase work efficiency.

On the other hand, it contains vitamin B12 (vitamin B12) and folic acid (folic acid) together with various kinds of amino acids to nourish cells and improve anemia.

Third, β-Pachymose (β-Pachimos), Pachymic acid, Lecithin, Lecithin, protein, fat, potassium, etc. contained in Bokryeong activate brain cells and stabilize the mind. It provides nutrition to improve weakness, and polysaccharides such as Pachyman and Pachymose are known to enhance immune function and exhibit antitumor activity. Choline promotes blood lipid degradation and is biosynthesized with Acethylcholine (acetylcholine) to activate brain cells.

Fourth, Catalpol (catalpol) contained in the live diabetes has a hypoglycemic action, Adenosine (adenosine) exhibits anti-thrombotic action, anti-rheumatic effect, iron with hematopoietic action to improve ischemia and promote renal blood flow diuretic Increase and relieve joint swelling and pain.

Fifth, 6-Gingerol (6-Gingerol) contained in Ginger inhibits prostaglandin synthesis, anti-ulcer, hepatotoxic detoxification, bile secretion and cardiac function, and Zingerol (Gingerol) strengthens the vascular locomotor center. It is known to promote blood circulation and promote digestion by promoting digestion. Shogaol has a hepatotoxic detoxification action, and Acetaldehyde has analgesic and mycelial action and diuretic effect by increasing renal glomerular blood flow. Furfural has a mucosal irritant effect and Caffeic acid has a hypolipidemic effect. Allylmethyl sulfide, allylmethyl sulfide, isopropyl sulfide, and diethyl sulfide, which are specific to ginger, are known to have antimicrobial activity.

Sixth, the control is rich in sugars, calcium, vitamin C, proteins, fats, etc. to nourish cells and activate metabolism. Ethyl-α-D-Fufuctofuranoside increases anti-allergic effect by increasing Cyclic AMP, and Nornuciferine has strong hypnosis and hardening effect. . Oxonuciferine has hypnotic action and C-AMP strengthens the mast cell membrane to show anti-allergic action and promote urine excretion. Maslinic acid has anti-inflammatory properties, inhibits histamine-induced bowel spasms, prevents stress ulcers, and exhibits antifungal activity.

In order to achieve the above object, a method for preparing an oral concentrated syrup containing a hydrolyzed extract of licorice and its complex herbal medicine is achieved by the following examples, and the constituent ratio corresponds to the present invention, Can be implemented in the range of ± 10%.

First embodiment;

To 400 grams of licorice, add 500 milliliters of 70% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50-60 ° C in a water bath, filter with filter paper, and collect the filtrate. 300 ml of purified water and enzyme solution 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃;

800 ml of 70% alcohol and 40 milliliters of 1% hydrochloric acid were added to 600 g of ginseng, warmed at 50-60 ℃ for 4 hours in a water bath, filtered through a filter paper, and the filtrate was collected. 400 ml of purified water and enzyme solution Adding 20 milliliters and fermenting at 28-32 ° C. for 30 days;

1.2 kg of 70% alcohol and 60 milliliters of 1% hydrochloric acid were added to 1.0 kg of Bokryeong, heated in a water bath at 50-60 ° C. for 4 hours, filtered through a filter paper, and the filtrate was taken. 800 ml of purified water and an enzyme solution were obtained. Adding 40 milliliters and fermenting at 28-32 ° C. for 30 days;

To 1.6 kg of raw sulfur, 2.2 liters of 70% alcohol and 110 milliliters of 1% hydrochloric acid were added and warmed in a water bath at 50-60 ° C. for 4 hours, filtered through a filter paper, and the filtrate was taken. Then, 1 liter of purified water and 50 milliliters of enzyme solution were added. Fermentation at 30 ° C. for 30 days;

To 400 grams of ginger, add 500 milliliters of 70% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50-60 ℃ in a water bath, filter with filter paper, filter the filtrate, and add 300 milliliters of purified water and 15 milliliters of enzyme solution. Fermentation at 30 ° C. for 30 days;

500 milliliters of 70% alcohol and 25 milliliters of 1% hydrochloric acid were added to 400 grams of control. The mixture was warmed at 50 to 60 DEG C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was collected. 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃;

Licorice, Ginseng, Bokyeong, Saenghwanghwang, Ginger, and the control solution, each extracted and fermented with alcohol solution from above, are mixed in one container and warmed at 80-90 ℃ for 4 hours in a water bath to completely volatilize the alcohol components, The concentrate was filled into an airtight container to obtain a final product.

Enzyme solution; Take 50,000 units of β-glucuronidase (betaglucuronidase) and add 100 ml of purified water.

Second embodiment;

To 400 grams of licorice, add 800 milliliters of 35% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50 ~ 60 ℃ in a water bath, filter with filter paper, take filtrate, add 15ml of enzyme solution, and add 30ml at 28∼32 ℃. Fermentation during;

To 600 grams of ginseng, 1.2 liters of 35% alcohol and 40 milliliters of 1% hydrochloric acid were added and warmed at 50 to 60 ° C. for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. Adding 20 milliliters and fermenting at 28-32 ° C. for 30 days;

2 kg of 35% alcohol and 60 milliliters of 1% hydrochloric acid were added to 1.0 kg of Bokryeong, warmed at 50 to 60 ° C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. Adding 40 milliliters and fermenting at 28-32 ° C. for 30 days;

To 1.6 kilograms of raw sulfur, 3.2 liters of 35% alcohol and 110 milliliters of 1% hydrochloric acid were added, warmed at 50 to 60 ° C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 50 milliliters of the fermentation for 30 days at 28 ~ 32 ℃;

To 400 grams of ginger, add 800 milliliters of 35% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50-60 ℃ in a water bath, filter with filter paper, take filtrate, add 15 milliliters of enzyme solution, and add 30ml at 28-32 ℃. Fermentation during;

To 400 grams of control, add 800 milliliters of 35% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50 ~ 60 ℃ in water bath, filter with filter paper, take filtrate, add 15ml of enzyme solution, and add 30ml at 28∼32 ℃. Fermentation during;

Licorice, Ginseng, Bokyeong, Saenghwanghwang, Ginger, and the control solution, each extracted and fermented with alcohol solution from above, are mixed in one container and warmed at 80-90 ℃ for 4 hours in a water bath to completely volatilize the alcohol components, The concentrate was filled into an airtight container to prepare a final product.

Third embodiment;

To 400 grams of licorice, 500 milliliters of purified water and 25 milliliters of 1% hydrochloric acid were added and warmed at 50 to 60 ° C. for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 300 milliliters of 95% alcohol and enzyme solution 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃;

To 600 grams of ginseng, 800 milliliters of purified water and 40 milliliters of 1% hydrochloric acid were added, warmed for 4 hours at 50-60 ° C in a water bath, filtered through a filter paper, and the filtrate was taken. 400 milliliters of 95% alcohol and enzyme solution Adding 20 milliliters and fermenting at 28-32 ° C. for 30 days;

1.2 kg of purified water and 60 milliliters of 1% hydrochloric acid were added to 1.0 kg of Bokryeong, warmed at 50-60 ° C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 800 ml of 95% alcohol and enzyme solution Adding 40 milliliters and fermenting at 28-32 ° C. for 30 days;

To 1.6 kilograms of fresh sulfur, 2.2 liters of purified water and 110 milliliters of 1% hydrochloric acid were added, heated at 50 to 60 ° C. for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 1 liter of 95% alcohol and an enzyme solution were added. 50 milliliters of the fermentation for 30 days at 28 ~ 32 ℃;

To 400 grams of ginger, 500 milliliters of purified water and 25 milliliters of 1% hydrochloric acid were added, warmed at 50 to 60 ° C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 300 milliliters of 95% alcohol and enzyme solution 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃;

To 400 grams of control, 500 milliliters of purified water and 25 milliliters of 1% hydrochloric acid were added, warmed at 50 to 60 ° C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 300 milliliters of 95% alcohol and enzyme solution 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃;

Licorice, Ginseng, Bokryeong, Saengjihwang, Ginger, and the control solution, which were extracted and fermented with purified water and alcohol solution from above, were all mixed in one container and warmed at 80-90 ° C. for 4 hours in a water bath to completely evaporate the remaining alcohol. The herbal complex concentrate was filled into an airtight container to prepare a final product.

Hereinafter will be described the action and experimental results of the extract or fermentation and hydrolyzate according to the present invention.

1. Identification and Quantification of Licorice and Its Hydrolysates

The contents of Glycyrrhizin (glycyrrhizin), the major component of licorice, and Glycyrrhetinic acid (glycyrrhitin acid), its hydrolyzate, were tested by HPLC analysis. The conditions were as follows.

Glycyrrhizin;

Column; Nucleosil C ₁₈ (4.0 × 250 mm)

Mobile phase; the mixture of HOH; CH ₃ CN; AcOH (560.0; 400.0; 40.0)

Flow rate; 1.2 mL / min

Injection volume; 20 μl

Detector; UV detector (λ _max = 254 nm)

Glycyrrhetinic acid;

Column; Spherisorb ODSI column C ₁₈ (4.0 × 250 mm)

Mobile phase; the mixture of HOH; CH ₃ CN; AcOH (170.0; 830.0; 5.0)

Flow rate; 1.2 mL / min

Injection volume; 20 μl

Detector; UV detector (λ _max = 254 nm)

Analysis of the contents of Glycyrrhizin and Glycyrrhetinic acid

HPLC analysis of the contents of Glycyrrhizin, a major component of licorice, and its hydrolyzate, Glycyrrhetinic acid, showed chromatograms of Glycyrrhizin at a wavelength of 245 nm. + 53.2, R² = 0.999 and good linearity up to 50 μg / mL.

[Figure 1] HPLC chromatogram of Glycyrrhizin

Column: Nucleosil C ₁₈ (4.0 × 250 mm)

Mobile phase: the mixture of HOH: CH ₃ CN: AcOH (560.0: 400.0: 40.0)

Flow rate: 1.2 mL / min

Injection volume: 20 μl

Detector: UV detector (λ _max = 254 nm)

On the other hand, the chromatogram of Glycyrrhetinic acid was shown at wavelength 245nm and the calibration curve was y = 74.95x-14.5, R² = 0.999.

[Figure 2] HPLC chromatogram of Glycyrrhetinic acid

Column: Spherisorb ODSI column C ₁₈ (4.0 × 250mm)

Mobile phase: the mixture of HOH: CH ₃ CN: AcOH (170.0: 830.0: 5.0)

Flow rate: 1.2 mL / min

Injection volume: 20 μl

Detector: UV detector (λ _max = 254 nm)

2. Principal Component Hydrolysis Experiment

1) Preparation and Measurement of Hydrolysates by Acid

The licorice extract was dissolved in pH 1, 3, 5, 7, 9 (90% ethanol) and then reacted at 40 ± 2 ° C for 2 days, and then the hydrolysis rate was measured for each hour.

2) Determination of hydrolysis using 1% HCl (hydrochloric acid), 1% NaOH (sodium hydroxide) and β-glucuronidase (β-glucuronidase)

Licorice extract was dissolved in 90% Ethanol, 1% HCl, 1% NaOH,

Hydrolysis rate was measured using β-glucuronidase (betaglucuronidase).

3) Measurement result of hydrolysis rate of main component

The increase rate of glycyrrhetinic acid by hydrolysis at pH 1.0 was 82.67% after 24 hours and 98.28% after 48 hours, higher than the group tested in other pH ranges. It was not done.

Table 1

Comparison of Hydrolysis Rate of Glycyrrhizin at Various pH Conditions in 90% Ethanol Solution at 40 ℃

pH1 pH3 pH5 pH7 pH9 24 hr 82.67 ±
2.41% 9.85 ±
2.51% 10.66 ±
2.55% 18.95 ±
1.98% 26.64 ±
1.37% 48 hr 98.28 ±
5.88% 25.44 ±
1.88% 13.88 ±
3.70% 45.30 ±
2.76% 73.04 ±
4.02%

Each data represents the mean ± SE from 5 experiments.

The hydrolysis rate using 1% HCl was 2-5 times higher than that of 1% NaOH and β-glucuronidase (1000 units).

[Table 2]

Comparison of Glycyrrhizin Hydrolysis Rate under Enzyme, Acid and Base Conditions in 90% Ethanol Solution at 40 ℃

B-glucuronidase 1% HCI 1% NaOH 24hr 61.25 ± 2.44% 66.49 ± 3.56% 12.5 ± 1.55% 48hr 85.23 ± 4.50% 180.28 ± 1.22% 15.75 ± 2.18% 72hr 118.78 ± 3.21% 240.09 ± 4.62% 24.57 ± 1.61%

Each data represents the mean ± SE from 5 experiments.

3. Antioxidant Experiment by DPPH Method

1,1-diphenyl-2-picrylhydrazil (1,1diphenyl-2-picrylhydrazyl, DPPH) was used for free radical scavenging activity of the main component and hydrogel. 1 mL of the solution diluted to various concentrations was added, shaken for 10 seconds with a vortex mixer, and allowed to stand at 20 ° C. for 30 min, and then absorbance was measured at 517 nm. Butyl hydroxy anisole (BHA) was used as a comparative standard. The magnitude of the activity is expressed in terms of Electron Donating Ability (EDA) for DPPH.

× 100EDA (%) =

× 100

Sample ABS; Absorbance of the test solution

Control ABS; Absorbance of Test Solution Added Methanol Instead of Sample

Antioxidant Effect Comparison

The free radical scavenging activity of the main component was BHA 70.41 ± 5.20% at 1000μg / ml concentration and licorice extract showed 55.67 ± 3.89% hydrogen donating ability and 55.05 ± 6.56 in licorice hydrolyzate extract. It did not show a big difference.

[Table 3]

Comparison of the Inhibitory Effects of Reactive Oxygen by DPPH Method

Sample EDA (1000 μg) EDA (100 μg) BHA
70.41 ± 7.25 67.56 ± 3.58 Licorice Extract
55.67 ± 3.89 38.11 ± 5.12 licorice
Hydrolysis extract
55.05 ± 6.56 32.27 ± 7.14

EDA; Hydrogen donating ability (%)

Each data represents the mean ± SE from 5 experiments.

4. Anti-inflammatory effect measurement experiment

1) Edema Suppression Experiment

The edema suppression experiment of hydrogel was conducted by applying the anti-inflammatory effect of carrageenan-induced rat foot edema. Edema was induced by dissolving 1% λ-carrageenan (λ-carrageenan) and 1% kaolin (kaolin) in PBS (phosphate buffered saline) at the bottom of the right foot of the rat. Drug application was applied once to the bottom of the right foot each 1 hour before induction.

Edema was measured using a plethysmometer and repeated three times at 6, 12, 24 and 48 hours before and after administration of 1% λ-carrageenan and 1% kaolin. The edema rate was calculated using the change of foot volume immediately after λ-carrageenan injection and at each time period. The edema rate (%, swelling) and edema inhibition rate (%, inhibition) were calculated by the following equation.

× 100 Edema rate (%) ⁼

× 100

] × 100Edema inhibition rate (%) = [1-

] × 100

V is the foot volume after 6, 12, 24 and 48 hours, V ₁ is the foot volume immediately after injection, and V _c is Edema rate of the control group, V _d is the edema rate of the treatment group.

2) Comparison of anti-inflammatory effects

When comparing the hydrogel edema and inhibitory effect by applying the treatment coefficient measurement method for carrageenan-induced edema, edema was the maximum at 2.3 mL in the control group 6 hours after carrageenan administration, and edema rate gradually after 12 hours. It began to decrease. Edema suppression effect of licorice extract began to appear after 12 hours, compared with 2.01 ± 0.03 ml in the control group, while 1.77 ± 0.04 mL in the control drug triamcinolone 1.80 ± 0.23 mL in the licorice extract, 1.75 ± 0.12 in the licorice hydrolyzate extract ml showed significant effect on the effect of suppressing edema.

Table 4

Inhibitory Effect of Licorice Hydrolyzate Extract on Carrageenan-Induced Foot Edema in Rats

Formulation
(Ml) Time (hr) 0 hr 6 hr 12 hr 24 hr 48 hr Control 1.14 ± 0.05 2.3 ± 0.08 2.01 ± 0.12 1.70 ± 0.05 1.35 ± 0.05 Licorice Extract 1.12 ± 0.03 2.06 ± 0.03 ^* 1.80 ± 0.04 1.49 ± 0.02 1.18 ± 0.07 Licorice Hydrolysis
extract 1.18 ± 0.08 2.11 ± 0.01 ^* 1.75 ± 0.06 ^* 1.40 ± 0.02 ^* 1.20 ± 0.06 ^*

Each data represents the mean ± SE from 5 experiments.

Statistically significantly value compared with control group data by T test ( ^* P <0.05, ^** P <0.01)

The edema rate of licorice extract, licorice hydrolyzate and triamcinolone decreased rapidly after 12 hours and the edema suppression effect was maintained until 48 hours.

In comparison, after 12 hours, the edema inhibition rate of the control group was only 20.44% in licorice extract, but 29.36% in triamcinolone and 36.7% in licorice hydrolyzate extract. While the X was 70.91%, the triamcinolone was 85.83% and the licorice hydrolyzed extract was 90.8%.

[Figure 3] Inhibitory Effect of Licorice Hydrolyzate Extract on Carrageenan-Induced Foot Edema in Rats

(GR: Licorice Extract, GHT: Licorice Hydrolyzate Extract, TA: Triamcinolone)

Each bar represents the mean ± SE from 5 experiments.

Statistically significantly value compared with GR group data by

Student's- t test ( ^* P <0.05, ^** P <0.01, ^*** P <0.001).

5. Analgesic effect measurement experiment

1) Heat pain threshold measurement experiment

The heat pain threshold was tested by applying a new heat pain threshold measurement method with increased sensitivity. First, a thermode (Turmod, Quck-Korea704D, Quick Electonic Equipment Factory, Korea) with a diameter of 2 mm of the probe tip was fixed at 50 ° C to avoid the reaction by lightly contacting the primary hyperalgesia and secondary hyperalgesia of rats. The time until this occurred was measured. The measurement was performed after adapting the rats for 10 minutes on a hot plate at 30 ° C for adaptation to the heat of the rats.

Measurements were performed before, 3 hours after induction, 6 hours after induction, and 9 hours after induction of hyperalgesia.

2) Mechanical pain threshold measurement experiment

Mechanical pain threshold measurement was performed by applying mechanical pain threshold measurement by acute compression method. First, the number of incidence of avoidance reflex in rats was measured using Von Frey filament (Bon Frey Filament, North Coast Medical Inc., Touch-Test Sensory Evaluation, USA). First, the rats were stimulated with low pressure using 1.4, 2, 4, 6, 8, 10, 15, and 26 g monofilament, to the primary and secondary hyperalgesia with minimal tenderness. The pressure was applied slightly to the tip of the monofilament. The number of evasion reflections was measured by applying a total of five times, and the interval between experiments was at least 2 minutes to prevent the adaptation of rats.

The measurement was conducted before, 3 hours after induction, 6 hours after induction, and 9 hours after induction of hyperalgesia.

3) Hazard flexion reflection test

The measurement of noxious flexion reflex was performed by applying the noxious flexion reflex measurement method by electrical nerve stimulation in inflammatory animal model. First, anesthetized rats with ether, followed by anesthesia, followed by anesthesia. Hazardous flexor reflex action potential was inserted into the femoral biceps of a rat using a pair of EL450 monopolar needles (Monopolar needle, Biopac system, USA). The EMG100C amplifier module (Amplifier module, Biopac system, USA) was used for recording. The ground electrode was attached to the tail.

Using a STIM100C stimulator module and STIMSOC (Seapac, Biopac system, USA), a pair of monopolar needles (monopolar needles) are inserted into the plantar feet of hyperalgesia. The timely pain threshold was induced, and then stimulated at 1.5 times the pain threshold and stored when the potential flexion reflex activity potential appeared on the EMG monitor.

The conditions for recording the activity potential were high pass filter, 500 Hz, low pass filter (low pass filter, 10 Hz, gain 1000 Hz, sample rate 1000 Hz, noise voltage) Tage) was 0.2 Hz and a notch-filter (notch filter) of 60 Hz was used, and the test stimulus was a single square wave with a 2 ms pulsation period.

The size of the reaction was calculated by RMS (root mean squre, mV). The recording and analysis of hazard flexion reflexes were carried out using the Acknowledge 3.8.1.Software program. Measurements were made before and after the onset of hyperalgesia.

4) Comparison of analgesic effects

Changes in the analgesia threshold were significantly different in all experimental groups compared to the control group. After 9 hours of induction, the control drugs of triamcinolone and licorice extract showed 25% analgesic inhibitory effect and 45% in licorice hydrolyzed extract. The best grade was shown in%.

[Figure 4] Analgesic Effects of Licorice Hydrolyzate Extracts on the Threshold of Threshold in Rats

(GR: Licorice Extract, GHT: Licorice Hydrolyzate Extract, TA: Triamcinolone)

Each bar represents the mean ± SE from 5 experiments.

Statistically significantly value compared with Blank group data by Student's- t test ( ^* P <0.05, ^** P <0.01).

After 6 hours of pain-induced change in mechanical pain threshold, significant differences were observed in all experimental groups compared to the control group. After 9 hours, triamcinolone, a control drug, showed about 20% pain suppression effect while 75 in licorice extract. Analgesic inhibitory effect was about%, and licorice hydrolyzed extract showed a much higher score (160%).

[Figure 5] Analgesic Effect of Licorice Hydrolyzate Extract on Mechanical Pain Threshold in Rats

(GR: Licorice Extract, GHT: Licorice Hydrolyzate Extract, TA: Triamcinolone)

Statistically significantly value compared with Blank group data by Student's- t test (* P <0.05, ** P <0.01).

Inflammatory animal models were 69.19 millivolts in the control group after 9 hours of change in adverse flexion response time due to electrical neurostimulation, 41.39 millivolts in the control triamcinolone, 40.15 millivolts in the licorice extract, and 40.46 millivolts in the licorice hydrolyzed extract All groups showed significant values, and the differences between the experimental groups were not significant.

[Table 5]

Analgesic Effect of Licorice Hydrolyzate Extract on Response Time of Hazardous Flexion Avoidance in Rats

pre 9h Blank 37.03 69.19 Licorice Extract 33.84 40.15 ^*** Triamcinolone 33.7 41.39 ^*** Licorice Hydrolyzate Extract 38.83 40.46 ^***

Each data represents the mean ± SE from 5 experiments.

Statistically significantly value compared with Blank group data by Student's- t test ( ^*** P <0.001).

On the other hand, the change in effective amplitude of harmful bend reflex was 0.45 seconds in the control group 9 hours after pain induction, while 0.41 seconds in triamcinolone, and 0.36 seconds in licorice extract and licorice hydrolyzed extract.

Table 6

Analgesic Effects of Licorice Hydrolyzate Extracts on the Amplitude Variation of Effective Value of Hazardous Flexion Avoidance in Rats

pre 9h Blank 0.41 0.45 Licorice Extract 0.35 0.36 ^* Triamcinolone 0.38 0.41 Licorice Hydrolyzate Extract 0.35 0.36 ^*

Each data represents the mean ± SE from 5 experiments.

Statistically significantly value compared with Blank group data by Student's- t test ( ^* P <0.05).

Claims (5)

To 400 grams of licorice, add 500 milliliters of 70% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50-60 ° C in a water bath, filter with filter paper, and collect the filtrate. 300 ml of purified water and enzyme solution 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃; 800 ml of 70% alcohol and 40 milliliters of 1% hydrochloric acid were added to 600 g of ginseng, warmed at 50-60 ℃ for 4 hours in a water bath, filtered through a filter paper, and the filtrate was collected. 400 ml of purified water and enzyme solution Adding 20 milliliters and fermenting at 28-32 ° C. for 30 days; 1.2 kg of 70% alcohol and 60 milliliters of 1% hydrochloric acid were added to 1.0 kg of Bokryeong, heated in a water bath at 50-60 ° C. for 4 hours, filtered through a filter paper, and the filtrate was taken. 800 ml of purified water and an enzyme solution were obtained. Adding 40 milliliters and fermenting at 28-32 ° C. for 30 days; To 1.6 kilograms of raw sulfur, 2.2 liters of 70% alcohol and 110 milliliters of 1% hydrochloric acid were added, heated at 50 to 60 ° C. for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 1 liter of purified water and an enzyme solution 50 milliliters of the fermentation for 30 days at 28 ~ 32 ℃; To 400 grams of ginger, 500 ml of 70% alcohol and 25 milliliters of 1% hydrochloric acid were added, warmed at 50 to 60 ° C. for 4 hours in a water bath, filtered through a filter paper, and the filtrate was collected. 300 ml of purified water and enzyme solution 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃; 500 milliliters of 70% alcohol and 25 milliliters of 1% hydrochloric acid were added to 400 grams of control. The mixture was warmed at 50 to 60 DEG C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was collected. 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃; Licorice, Ginseng, Bokyeong, Saenghwanghwang, Ginger, and the control solution, each extracted and fermented with alcohol solution from above, are mixed in one container and warmed at 80-90 ℃ for 4 hours in a water bath to completely volatilize the alcohol components, A method for producing a concentrated syrup for internal use containing a hydrolyzed extract of licorice and the complex herbal, characterized in that the concentrate is filled into an airtight container to produce a final product. The method of claim 1, The enzyme solution β-glucuronidase (betaglucuronidase) A method for producing concentrated syrup for internal use containing hydrolyzed extracts of licorice and its complex herbal medicine, characterized in that it is prepared in 100 milliliters by taking 50,000 units (titer) of purified water. To 400 grams of licorice, add 800 milliliters of 35% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50-60 ℃ in a water bath, filter with filter paper, and take the filtrate. 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃; Add 1.2 liters of 35% alcohol and 40 milliliters of 1% hydrochloric acid to 600 grams of ginseng, warm for 4 hours at 50-60 ℃ in a water bath, filter with filter paper, take filtrate, add 20 milliliters of enzyme solution, and add 30 ml at 28-32 ℃ for 30 days. Fermentation during; 2 kg of 35% alcohol and 60 milliliters of 1% hydrochloric acid were added to 1.0 kg of Bokryeong, warmed at 50 to 60 ° C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. Adding 40 milliliters and fermenting at 28-32 ° C. for 30 days; To 1.6 kilograms of raw sulfur, 3.2 liters of 35% alcohol and 110 milliliters of 1% hydrochloric acid were added and warmed at 50 to 60 ° C for 4 hours in a water bath, filtered through a filter paper, filtrate was added 50 ml of enzyme solution, and 30 days at 28 to 32 ° C. Fermentation during; To 400 grams of ginger, add 800 milliliters of 35% alcohol and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50-60 ℃ in a water bath, filter with filter paper, take filtrate, add 15 milliliters of enzyme solution, and add 30ml at 28-32 ℃. Fermentation during; To 400 grams of control, 800 milliliters of 35% alcohol and 25 milliliters of 1% hydrochloric acid were added, warmed at 50 to 60 ° C. for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 15 milliliters of fermentation for 30 days at 28 ~ 32 ℃; Licorice, Ginseng, Bokyeong, Saenghwanghwang, Ginger, and the control solution, each extracted and fermented with alcohol solution from above, are mixed in one container and warmed at 80-90 ℃ for 4 hours in a water bath to completely volatilize the alcohol components, A method for producing a concentrated syrup for internal use containing a hydrolyzed extract of licorice and the complex herbal, characterized in that the concentrate is filled into an airtight container to produce a final product. The method of claim 1, The enzyme solution β-glucuronidase (betaglucuronidase) A method for producing concentrated syrup for internal use containing hydrolyzed extracts of licorice and its complex herbal medicine, characterized in that it is prepared in 100 milliliters by taking 50,000 units (titer) of purified water.  To 400 grams of licorice, add 500 milliliters of purified water and 25 milliliters of 1% hydrochloric acid, warm for 4 hours at 50-60 ° C in a water bath, filter with filter paper, filter the filtrate, add 300 milliliters of 95% alcohol and 15 milliliters of enzyme solution. Fermentation at 30 ° C. for 30 days; Add 800 milliliters of purified water and 40 milliliters of 1% hydrochloric acid to 600 grams of ginseng, warm it for 4 hours in a water bath at 50-60 ℃, filter it with a filter paper, take a filtrate, add 400 milliliters of 95% alcohol and 20 milliliters of enzyme solution. Fermentation at 30 ° C. for 30 days; 1.2 kg of purified water and 60 milliliters of 1% hydrochloric acid were added to 1.0 kg of Bokryeong, warmed at 50-60 ° C for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 800 ml of 95% alcohol and enzyme solution Adding 40 milliliters and fermenting at 28-32 ° C. for 30 days; To 1.6 kilograms of fresh sulfur, 2.2 liters of purified water and 110 milliliters of 1% hydrochloric acid were added, heated at 50 to 60 ° C. for 4 hours in a water bath, filtered through a filter paper, and the filtrate was taken. 1 liter of 95% alcohol and an enzyme solution 50 milliliters of the fermentation for 30 days at 28 ~ 32 ℃; To 400 grams of ginger, 500 milliliters of purified water and 25 milliliters of 1% hydrochloric acid were added, heated at 50-60 ° C. for 4 hours in a water bath, filtered through a filter paper, and filtrate was collected. 300 ml of 95% alcohol and 15 milliliters of enzyme solution were added. Fermentation at 30 ° C. for 30 days; To 400 grams of control, 500 milliliters of purified water and 25 milliliters of 1% hydrochloric acid were added, warmed at 50 to 60 ° C for 4 hours in a water bath, filtered through a filter paper, and filtrate was taken. 300 ml of 95% alcohol and 15 milliliters of enzyme solution were added. Fermentation at 30 ° C. for 30 days; Licorice, Ginseng, Bokryeong, Saengjihwang, Ginger, and the control solution, which were extracted and fermented with purified water and alcohol solution from above, were all mixed in one container and warmed at 80-90 ° C. for 4 hours in a water bath to completely evaporate the remaining alcohol. A method for producing a concentrated syrup for internal use containing a licorice and a hydrolyzed extract of the complex herbal, characterized by filling the herbicidal complex concentrate in an airtight container to produce a final product.