KR100441158B1 - The functional drinks containing Crataegi fructus extracts and process for preparation thereof - Google Patents

Abstract

The present invention relates to a functional beverage containing a mountain lion extract and a method for preparing the same, which are prepared in a powder form by concentrating the hot water extract or ethanol extract of the mountain lion, and then examined its physiological activity and has high antioxidant activity, whitening effect, and angiotensin The inhibitory activity of the converting enzyme was excellent. Therefore, the functional beverage containing the sansa extract of the present invention has a high antioxidant activity, a whitening effect, and has an excellent effect of preventing hypertension.

Description

The functional drinks containing Crataegi fructus extracts and process for preparation, etc.

The present invention relates to a functional beverage containing a mountain lion extract and a method for preparing the same, and more particularly, the present invention relates to the production of a functional beverage containing a mountain lion extract having high antioxidant activity, whitening effect and effective in preventing hypertension. will be.

Mountain Lion ( Crataegi fructus ) is a fruit of Sanryong and Crataegus pinnatifida ( BUNGE) , which grows in wild fields and valleys in Korea. It is a deciduous and deciduous arbor tree, also known as a locust tree, grows about 6m in height and has sharp thorns on its branches. White flowers of about 1.5-1.8 cm in diameter bloom in May-June. The hawthorn, also called agawi, is a ball-like nucleus and ripens in September-October, and is a small apple-like sphere with a diameter of 1.5- 2.5 cm, scattered with small white spots. In general, it is made of colorful or eaten raw, and is often drunk. Sun-dried fruit is called Sansa ( Crataegi Fructus ) and is used as a moisturizer and medicinal herb in oriental medicine. It is also eaten as a digestive agent after eating meat. Jusanji is found in many dry, sandy and stony dry mountainsides around the southern part of China's transfiguration. In Japan, it is cultivated as a bonsai in parks and gardens, and is also called mayflower in foreign countries.

In Korea, the hawthorn, Crataegus maximowiczii , birch ( C. pinnatifida f. Betulifolia ), broadleaf hawthorn ( C. pinnatifida var. Major ), and brash hawthorn ( C. pinnatifida var. Partita 7 kinds of wild hawthorn, wild jasminoides , C. pinnatifida var. Psilosa and C. pinnatifid var. Typica . After harvesting and drying the mature fruits of these plants, they are called hawthorn ( Crataegi Fructus ) in Chinese herbal medicine ( Buksansa (fruit of C. pinnatifida var. Major ) native to China, Namsansa (fruit of C. cuneata ) Digestion, formal medicine, etc. are used a lot. Fruits such as C. oxyacantha are also used in Europe.

In China, mountain lions are used as herbal medicines and herbal medicines, but they are also used as candy and medicinal ingredients, and as foods closely related to people's lives such as jelly.

Sansa's weakness and medicinal properties are lukewarm, non-toxic and subtractive in taste, while the ear is secreted in the stomach, stomach and liver. In particular, mountain breeding (山査 肉) is news (소식 食 積), uric acid health (위 健胃), toothy (治 肉 積), mountain fish blood (疝 瘀血), structural insects (驅 條 蟲) It is said to be used for the treatment of jinga, soak, obesity, carbonic acid, saree, jangfeng, postpartum morning pain, obesity, pediatric stagnation, dysmenorrhea and low back pain. In particular, good health, digestion, astringent, analgesic, sterilization, insecticides are known to be good, and is said to be good for hangovers. In contrast, European Sansa leaves and flowers extracts are used for the treatment of circulatory diseases such as hypertension, angina, arrhythmia, coronary atherosclerosis and hypercholesterolemia.

Pharmacological components of mountain lions include ursolic acid, quercetin, crataegolic acid, tartaric acid, and hyperside vitexin-4-rhamnoside. Citric acid, caffeic acid, choline, ascorbic acid (0.089%), carbohydrate (22%), protein (0.7%), fat (0.2%), lipase, etc. It is contained.

Recently, research on the physiological activity of natural products has been actively conducted. Kim et al. Found that the antioxidative effect of medicinal herb extracts was superior to synthetic antioxidants.Phenol-based compounds, which are antioxidant components isolated from natural products, have hydroxyl groups as free radical acceptors among free oxides. Since it forms a stable hybrid in a step (resonance hybrid) is known to act as an antioxidant. The phenolic constituents of the acidic acid are caffeic acid, protocatechuic acid and pyrogallol in the free phenolic acid of the acidic acid, caffeic acid in the soluble ester type phenolic acid, phloroglucinol, insoluble phenols. Protocatechuic acid and prologlucinol were identified in the acid, confirming that they are the main antioxidants of the hawthorn extract.

Currently, most clinical studies, pharmacological studies, and medicinal components of the midwifery have been conducted. However, studies on the enzymatic aspects of physiological activity of Korean midwifery have not progressed much. The development of commercialization has not been done so far.

Accordingly, an object of the present invention is to verify the physiological activity of Korean hawthorn, which is widely used as a herbal medicine, and to provide a functional drink containing hawthorn extract and a method of manufacturing the same.

The above object of the present invention is obtained by lyophilization by hot water extraction and ethanol extraction of Korean hawthorn, and then obtained in a powder state to investigate the antioxidant activity by measuring the electron donating ability and superoxide dismutase activity by measuring the tyrosinase inhibitory activity The anti-hypertensive effect was investigated by investigating the whitening effect and the inhibitory activity of angiotensin converting enzyme, and then preparing the functional beverage containing the hawthorn extract by adding hawthorn fragrance, honey, vitamin C, citric acid and fructose to the hawthorn extract obtained above. By achieving this.

Figure 1 is a schematic of the process of analyzing the electron donating ability of the hawthorn extract of the present invention.

2 is a graph showing the results of the electron donating ability of the hawthorn extract of the present invention.

Figure 3 illustrates the process of measuring the similar activity of the superoxide dismutase (SOD) of the mountain lion extract of the present invention.

Figure 4 shows the result of measuring the similar activity of the superoxide dismutase (SOD) of the mountain lion extract of the present invention.

Figure 5 is a schematic of the process for measuring the tyrosinase inhibitory activity of the hawthorn extract of the present invention.

Figure 6 shows the result of measuring the tyrosinase inhibitory activity of the mountain lion extract of the present invention.

Figure 7 illustrates the process of measuring the angiotensin converting enzyme (ACE) inhibitory activity of the hawthorn extract of the present invention.

8 is a graph showing the results of measuring the angiotensin converting enzyme (ACE) inhibitory activity of the hawthorn extract of the present invention.

The present invention comprises the steps of adding hot water extract and ethanol extract by adding distilled water to the Korean mountain lion; Lyophilizing the extract to concentrate and powder; Investigating the electron donating ability and superoxide dismutase activity, tyrosinase inhibitory activity and inhibitory activity of angiotensin converting enzyme of the hawthorn extract; Comprising the production of a beverage by adding the hawthorn fragrance, honey, vitamin C, citric acid, fructose to the hawthorn extract.

Hereinafter, the specific configuration and operation of the present invention will be described with reference to Examples, but the scope of the present invention is not limited only to these Examples.

EXAMPLE

Example 1 Preparation of Hawthorn Extract

Preparation of Hawthorn Extract

The mountain lion ( Crataegi Fructus ) used in the present invention was purchased after washing Korean water from Daegu Yangnyeong Market and dried in the shade. The extraction of the sample was performed by reflux extraction at 80 ° C. for 3 hours by adding 10 times the amount of distilled water to 100 g of the sample. The supernatant and sediment were separated, the supernatant was collected separately, and the precipitate was extracted once and repeatedly. Ethanol extraction of the sample was added to the 100g sample 10 times the amount of 70% ethanol and extracted for 24 hours at room temperature to separate the supernatant and sediments were extracted once again in the same manner as above. Each extracted supernatant was centrifuged, filtered, concentrated, lyophilized, stored in a freezer, and used as a sample of the present invention.

Reagents and Instruments

Reagents used in enzyme experiments to measure the physiological activity of hawthorn are DPPH (Sigma), Xanthine (Sigma), XOase (Sigma), Pyrogallol (Sigma), L-Dopa ( Sigma company), tyrosinase (Sigma company), rabbit lung acetone powder (Rigbit lung acetone powder, Sigma company), HHL (Sigma company) and the like were used.

The MTT assay used in experiments to investigate the effects of hawthorn extracts on the proliferation of melanoma cells was performed using RPMI 1640 medium (Gibcosa), fetal bovine serum (Sigma), penicillin / streptomycin (Gibco Co., Ltd.), MTT (Sigma Co., Ltd.), Trypsin 250 (Diffco Co., Ltd.) was used, Trypan blue stain 0.4% (Trypan blue stain 0.4%, Gibco Co., Ltd.) and hemasytometer (haemacytometer, Marien) Field company) was used. Other reagents were used as express reagents.

Instruments used in the present invention include UV / vis spectrophotometer (Hitachi Co., Ltd.), rotary vacuum evaporator (Tokyo Rikakikai Co., Ltd.), centrifuge (Hitachi Co., Ltd.), lyophilizer , Optical microscope (Olympus), CO ₂ incubator (Hanbaek Scientific), pH meter (Orion Research), BOD incubator (Hanbaek Scientific), pressurized steam sterilizer (Hanbaek Scientific), ELISA reader It measured using (Biorad Corporation make).

Example 2: Determination of the biological activity of the hawthorn extract

Experimental Example 1 Measurement of Electron Donating Ability of Sansa Extract

Free radicals occur as biological reactions during metabolism by flavoenzymes such as xanthine oxidase or glutathione reductase in the mitochondria, phagocytes or cytoplasm, and electron donating acts to provide electrons to these oxidative free radicals Inhibits oxidation. Therefore, the electron donating ability was investigated to determine the degree of antioxidant inhibition of the hawthorn extract. Electron donating abilities (EDA) of the hawthorn extracts were measured according to FIG. 1 by modifying the method of Blois. Each sample solution 2.0mL 2 × 10 ^-4 M of α, α- diphenyl picryl -β- - After stirring Hyde Brazilian (DPPH) into 1.0mL was allowed to stand for 30 min and then the absorbance was measured at 517nm. The electron donating ability was expressed as the absorbance reduction rate of the sample solution addition and no addition solution.

S _Abs : absorbance of the sample measured at 517 nm

B _Abs : absorbance of distilled water measured at 517 nm instead of DPPH

C _Abs : absorbance of distilled water measured at 517 nm instead of sample

As shown in FIG. 2, as a result of measuring the DPPH radical scavenging activity of each extract, the electron donating ability was increased as the concentration of the extract increased, and the hot water extract and the ethanol extract of 100 ppm of hawthorn were 29.5% and 35.7%. The electron donating ability of showed high electron donating ability of 89.4% and 91.8% at 500ppm concentration, respectively. In addition, the concentration of 1,000 ppm showed electron donating ability of 92% and 97.8%. It was found that the hawthorn extract of the present invention has a high electron donating ability as compared to the synthetic antioxidant BHT showed an electron donating ability of 26.3%, 44.8%, 66.7% at the concentration of 100ppm, 500ppm and 1,000ppm, respectively.

Experimental Example 2: Measurement of the similar activity of the superoxide dissolves (SOD)

Superoxide dissolvease is a natural antioxidant that oxidizes superoxide radicals into oxygen in vivo. It is known as a protein or enzyme that converts active oxygen species into stable substances and protects living organisms from oxygen injury. Therefore, SOD-like activity, which is known to be closely related to anti-oxidation as well as anti-ageing, was measured in the extracts of the hawthorn and investigated by the automatic oxidation of pyrogallol. SOD-like activity was measured by the method of FIG. 3 according to Marklund's method. To 0.2 mL of each sample solution, 3.0 mL of Tris-HCl buffer solution (50 mM tris + 10 mM EDTA, pH 8.5) and 0.2 mL of 7.2 mM pyrogalol were added and reacted at 25 ° C. for 10 minutes, followed by reaction with 1.0 N HCl 0.1 mL. The amount of pyrogallol oxidized in the reaction solution was stopped and measured at 420 nm. The similar activity of SOD was shown by the absorbance reduction rate of the sample solution addition group and no addition group.

S _Abs : absorbance of the sample measured at 420 nm

B _Abs : absorbance of buffer measured at 420 nm instead of pyrogallol

C _Abs : absorbance of buffer measured at 420 nm instead of sample

As shown in Figure 4, as a result of examining the SOD-like activity of each extract of the mountain lion of the present invention by concentration, the hot water extract and ethanol extract showed a slight similar activity of 19.6% and 24.5% at 5,000ppm, but as the concentration increased Activity increased.

Experimental Example 3: Determination of Tyrosinase Inhibitory Activity of Sansa Extract

Melanin is a natural pigment of a polymer widely found in animals, plants and microorganisms. Various kinds of melanin are known according to organisms, and are produced through multi-step processes such as enzymatic and non-enzymatic oxidation and polymerization of phenols. The mechanism of tyrosinase inhibitory activity may be a very important part in the prevention of browning in the food industry, especially the whitening effect of the cosmetic industry.

The in vivo melanin synthesis process is a continuous process of first producing L-3.4-dihydroxyphenylalanine using tyrosine as a substrate and transferring it back to L-dopaquinone. After enzymatic oxidation (hydroxylation and oxidation) proceeds by polymerization of each product.

Sanshima et al. See two major types of approaches to suppress melanogenesis. One type is a type that directly inhibits tyrosinase enzyme itself, which is a major enzyme of melanin production, and the other type is a type that suppresses melanin production in skin pigment cells, although there is no direct inhibition on tyrosinase isolated from cells. to be.

The present invention is to see the inhibitory effect of the enzyme activity on the electronic type, the measurement of the inhibitory activity of tyrosinase according to the method of Yagi et al. To measure the dopa chrome produced by the action of tyrosinase by colorimetric method. It was investigated according to the method of FIG. To the mixture of 0.2 mL of 10 mM L-DOPA, 0.2 mL of 0.175 M phosphate buffer (pH 6.8) and 0.5 mL of sample solution, 0.1 mL of tyrosinase (110 units / mL) of mushroom was added and reacted at 35 ° C. for 2 minutes. Dopa chromium produced in the solution was measured at 475 nm, and the inhibition rate was calculated by the following equation.

S _Abs : absorbance of the sample measured at 475 nm

B _Abs : absorbance of buffer measured at 475 nm instead of enzyme

C _Abs : absorbance of buffer measured at 475 nm instead of sample

As shown in Figure 6, by using the tyrosinase of mushrooms to measure the inhibitory effect of tyrosinase activity of each extract of the mountain lion of the present invention by concentration, the hot water extract and ethanol extract of the Although there was no inhibitory effect, high concentrations of 49.8% and 62.4% were inhibited at concentrations of 30,000 ppm, respectively. In addition, it was found that at 50,000 ppm, enzyme activity was inhibited in a concentration-dependent manner, showing high inhibitory activity at 82.8% and 96.7%, respectively.

Experimental Example 4 Measurement of Inhibitory Activity of Angiotensin Converting Enzyme (ACE)

In the mechanism of hypertension, the renin-angiotensin system plays an important role in blood pressure control. In particular, angiotensin converting enzyme is involved in the final step of synthesizing angiotensin II, an octapeptide having strong vasoconstrictive action by hydrolyzing C-terminal double peptide from angiotensin I, a decapeptide produced by renin. It is an enzyme. Angiotensin II produced has strong vasoconstrictive action and inhibits excretion of water and sodium by promoting the release of aldosterone from the adrenal cortex. ACE also inactivates bradykinin, a nonapeptide with vasorelaxant action, resulting in elevated blood pressure.

Therefore, in order to measure the inhibitory activity of the ACE of the hawthorn extract of the present invention, a modification of Cushman et al. Was carried out according to the method of FIG. That is, the experimental group was dissolved 0.15mL of substrate (Hippuryl-L-histidyl-L-leucine: HHL) in 0.1M potassium phosphate buffer (pH 8.3) containing 0.3M NaCl, rabbit lung acetone powder (Rabbit lung acetone powder) 0.1mL and 0.1mL of the sample solution were mixed, and the control group was added 0.1mL of distilled water instead of the sample solution and reacted at 37 ° C for 30 minutes. The reaction was stopped by adding 0.35mL of 1N HCl, and then 3mL of ethyl acetate was added. 1 mL of distilled water was added to the residue from which the solvent was distilled from the ethyl acetate layer, and the extracted hyperfuric acid was measured at absorbance of 228 nm, and the inhibition rate was determined according to the following equation.

S _Abs : absorbance of the sample measured at 228 nm

B _Abs : absorbance of distilled water measured at 228 nm instead of enzyme

C _Abs : absorbance of distilled water measured at 228 nm instead of sample

As shown in Figure 8, the inhibition of the ACE enzyme activity of the hawthorn extract of the present invention showed the inhibitory activity of 32.7% and 34.5% at a concentration of 10ppm hawthorn extract, respectively, but the inhibitory activity is great at increasing the concentration There was no change and ACE inhibitory activity was higher in hot water extract than ethanol extract. As a result, the hawthorn extract showed a high activity at a low concentration is a noteworthy result.

Example 3 Preparation of Functional Drink Containing Sansa Extract

The mountain lion extract of the present invention obtained the mountain lion extract according to the preparation method of Example 1 and examined its physiological activity through Example 2 it was confirmed that the antioxidant activity is high, whitening effect and high blood pressure prevention effect. Thus, in order to prepare a functional beverage containing the mountain lion extract having the above effect, according to the method of Example 1 to obtain a mountain lion extract in the powder form of the lioness extract 0.01 ~ 10% by weight of the mountain lion flavor 0.08% by weight, honey 0.95 It was prepared by adding 0.1% by weight, 0.1% by weight of vitamin C, 0.1% by weight of citric acid, and 14% by weight of fructose.

As described above, the present invention, as described through the examples, the hot water extract and ethanol extract of the mountain lion of the present invention has high electron donating ability, high antioxidant activity, high activity of superoxide dismutase related to scavenging of radicals, and whitening. It is effective and has an excellent effect of inhibiting the activity of angiotensin converting enzyme. Therefore, the functional beverage containing the hawthorn extract of the present invention is a useful invention in the functional beverage manufacturing industry because it has a high antioxidant activity, has a whitening effect, and has an excellent effect on preventing hypertension.

Claims (2)

10 times of distilled water was added to the hawthorn, and reflux-cooled at 80 ° C. for 3 hours. The supernatant and the precipitate were separated to obtain a supernatant. The supernatant was centrifuged and filtered to obtain a filtrate. Prepare powder, 0.01 to 10% by weight of the powdered lioness extract, 0.08% by weight of lioness scent, 0.95% by weight of honey, 0.1% by weight of vitamin C, 0.1% by weight of citric acid, characterized in that it comprises the production by adding 14% by weight fructose Method for producing a functional beverage containing a mountain lion extract. A functional beverage prepared by the method according to claim 1, which contains SOH-like activity, high whitening effect, active oxygen removing ability, delaying aging and hypertension prevention as an active ingredient.