KR20070084933A - Antioxidant-substance extracted from cucumber plants - Google Patents

Abstract

An extract of Cucumis sativus L. is provided to remove the toxicity of paraquat completely and show excellent active oxygen species(such as hydrogen peroxide, superoxide anion and hydroxyl radical) scavenging activity, thereby being used for an antioxidant composition. The antioxidant composition comprises an extract of Cucumis sativus L. as an effective ingredient. The method for preparing the extract comprises the steps of: (a) extracting Cucumis sativus L. with at least one solvent selected from the group consisting of methanol, ethanol, propanol, and chloroform and concentrating it under reduced pressure; (b) respectively fractionating the concentrate with hexane, ethyl ether, ethyl acetate, butanol and water; and (c) performing silica gel column chromatography, Sep-Pack C18 cartridge and HPLC in sequence on fractions obtained from the step(b).

Description

Cucumber Plants with Cucumber Herbal Extract with Antioxidant Activity

1 shows detoxification activity against fractions and paraquats of cucumber herbal extracts.

Figure 2 shows the effect of inhibiting the leakage of cell membrane electrolyte by paraquat cucumber extract (water soluble fraction).

Figure 3 shows the effect of inhibiting the production of malondialdehyde by paraquat cucumber extract (water soluble fraction).

Figure 4 shows the antioxidant activity of cucumber herb extract (aqueous fraction) in scavenging 1,1-diphenyl-2-picrylhydrazil compared to ascorbic acid, alphatocopherol, resveratrol.

Figure 5 shows the antioxidant activity of cucumber herb extracts (aqueous fractions) scavenging hydrogen peroxide.

Figure 6 shows the antioxidant activity of cucumber herb extracts (aqueous fractions) scavenging superoxide anions.

The present invention relates to a cucumber herbal extract having an antioxidant activity, and more particularly, Cucumis sativus L. herbal (young leaf) extract completely removes the toxicity of paraquat, hydrogen peroxide (hydrogen peroxide), superoxide anion ( The present invention relates to an antioxidant composition containing a cucumber herbal extract by revealing a strong antioxidant activity with excellent scavenging activity of active oxygen species such as superoxide anion) and hydroxyl radical.

In addition to hydroxyl radicals, free radicals and reactive oxygen species or reactive nitrogen species and peroxynitrite (ONOO-) are well known to play an important role in causing a wide variety of human diseases. 1,1-diphenyl-2-picrylhydrazyl (DPPH) is a free radical compound and has been widely used to test the free radical scavenging ability of various chemicals. Peroxynitrite, the result of the reaction of superoxide and nitric oxide, is a species that destroys cells by oxidizing some cellular components such as proteins, lipids, and DNA. It is known to be associated with diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. In addition, reactive oxygen species (ROS) are closely associated with various degenerative disorders associated with aging. Active oxygen species, such as hydrogen peroxide, superoxide anion, hydroxyl radicals, and nitric oxide, can cause aerobic metabolism and damage all components of the cell, such as nucleic acids, proteins, and lipids. Is generated. Various efforts have been made to prevent oxidative damage caused by the various radicals, and in particular, there is a growing interest in natural compounds having antioxidant activity. Among the antioxidants, antioxidants that inhibit lipid peroxidation play an important role in not only preserving food but also protecting oxidative damage of living cells. Many natural products found in plants are known to retard the oxidation process in natural environments as scavengers of free radicals and free radicals, and in experiments as reducing agents.

In the body, superoxide dismutase (SOD) plays an important role in protecting somatic cells from free radicals. Alzheimer's disease is reported to be caused by an imbalance between free radicals and antioxidants in somatic cells. Types of dementia include cerebrovascular dementia caused by heart disease, high blood pressure, and arteriosclerosis, and Alzheimer's disease with unknown causes. Symptoms of Alzheimer's dementia occur in the order of dysentery, confusion, memory impairment, and complications of dementia, resulting in a sharp decrease in brain neurons leading to death. The cause of Alzheimer's dementia has not yet been accurately identified. The following hypothesis explains the cause of dementia. The first hypothesis is attributed to the abnormality of the 21st chromosome of amyloid (beta protein) involved in precursor formation, and the other hypothesis is caused by mutation of some amino acids forming amyloid. If the disease occurs early in the old age, the 19th chromosome abnormalities are said to develop dementia after age 65. Some hypotheses are caused by the accumulation of a large amount of aluminum in the neurofibrillary tangles, and some are due to the imbalance between free radicals and antioxidants and antioxidant enzymes.

There is no specific treatment for Alzheimer's disease.Tacrine, a substance that activates the action of acetylcholine, has been reported in the United States, and L-Cartinine, a potent antioxidant, increases levels of acetylcholine. Was also reported. In addition, saponin extract is manufactured and sold as a treatment for dementia, and many drugs are being tested, but the clinical effect has not been recognized. On the other hand, inflammation, lung disease, arthritis, etc. can also occur depending on the conditions of the active oxygen, and research on the role of SOD as a countermeasure against this.

Paraquat (1,1-dimethyl 4.4'-dipyridium dichloride) is a fast-acting herbicide that has been used most widely in the world, including Korea, since herbicides have been discovered since the mid-1950s. The mechanism is being studied extensively. When paraquat is exposed to the human body, gastrointestinal disorders, hepatic insufficiency, and renal injuries due to local stimulation appear. Finally, lung injuries appear, leading to death by lung fibrosis and kidney failure. In humans, the mortality of paraquat is 4 mg / kg and the half-lethal concentration (LD50) is 120 mg / kg. Paraquat-induced pulmonary toxicity represents the ideal of the destructive and proliferative phases. At the beginning of the destruction phase, the membrane is completely exposed to facilitate the secondary collapse of the basement membrane of the alveolar, and the most characteristic pulmonary edema of the destruction phase appears, which progresses to the regeneration phase, and the lung rapidly becomes fibroblasts. Differentiate and infiltrate with early fibroblasts and sometimes progress to fibrosis. When a single high dose of paraquat is administered to an animal, ultrastructural lesions are first observed in alveolar epithelial cell type I of alveolar epithelial tissues, swelling of the vesicles and mitochondria, and mitochondrial water. Increasing vesicles and thick vesicles appear in the cytoplasm and necrosis occurs in the basal membranes exposed on type I cells when administered at higher doses than half lethality. Paraquat's pulmonary toxicity mechanism is due to the in vivo interaction of paraquat radicals produced by the redox cycle with NADPH-dependent cytochrome P450 reductase and xanthine oxidase. The production of free radicals, superoxide anion and hydroxyl radicals, results in cell membrane damage, degradation of hyaluronic acid, inactivation of proteins, DNA damage and NADPH due to lipid peroxidation. Inhibition of fatty acid biosynthesis by hydrogen desorption occurs. In addition, as in the mechanism of action of quinone-based anticancer drugs, the reduction of intracellular NADPH causes a reduction of reduced glutathione, which promotes lipid peroxidation and causes disorders. Paraquat stimulates alveoli in the lungs to induce inflammation, stimulates fibroblasts in lung tissues to promote fibronectin, collagen biosynthesis, and cause pulmonary fibrosis. do. Paraquat is currently used for indirect concentration or direct handling carelessness and suicide for death, but there are no effective treatments and treatments. Commonly used therapies and treatments include adsorbents and activated charcoal to adsorb and remove paraquat in the gastrointestinal tract, and hemodialysis, hemofusion, and forced diuresis for the purpose of promoting the excretion of paraquat. . In particular, in order to reduce pulmonary toxicity of paraquat, spermine, a competitive paraquat inhibitor, is used in consideration of the use of a receptor of an amine compound such as putrescine in the process of paraquat inflow into the lung. Polyamine compounds such as spermidine, etc. are used, and various studies have been conducted to block the main metabolic system of oxygen radical generation, and steroids such as glucocorticoid, an anti-inflammatory agent, Steroid preparations, immunosuppressants, B-blockers, vitamin C, tocopherol and superoxide dismutase (SOD) have been studied.

Thus, the present inventors have conducted a study on the antioxidant activity of the cucumber herb extract has a function of completely detoxifying paraquat treated in plants, malondialdehyde (MDA) measurement, electrolyte leakage, soybean It was confirmed through leaf biotest, and the scavenging activity of active oxygen species such as hydrogen peroxide, superoxide anion, hydroxyl radical, etc. was examined to show that the extract of cucumber herb has strong antioxidant activity. The present invention was completed by revealing.

Accordingly, it is an object of the present invention to provide an antioxidant composition and paraquat antidote containing Cucumis sativus L. herbal extract as an active ingredient.

In addition, another object of the present invention to provide a pharmaceutical, health food, cosmetics, pesticides, etc. containing a cucumber ( Cucumis sativus L.) herbal extract as an active ingredient.

The present invention is characterized by an antioxidant composition containing Cucumis sativus L. herbal extract as an active ingredient.

In another aspect, the present invention is characterized by another paraquat antidote containing Cucumis sativus L. herbal extract as an active ingredient.

In addition, the present invention includes the use of medicines, health foods, cosmetics, pesticides and the like containing Cu ( cucumis sativus L.) herbal extract as an active ingredient.

The present invention will be described in more detail as follows.

In the present invention, Cucumis sativus L. Herbal (young leaf) extract completely eliminates the toxicity of paraquat, active oxygen species such as hydrogen peroxide, superoxide anion, hydroxyl radical, etc. The present invention relates to an antioxidant composition containing a cucumber herbal extract by showing an excellent scavenging activity and having a strong antioxidant activity.

First, the cucumber herbal extract according to the present invention is obtained through the following process.

Cucumber herb (young leaves) is extracted with one or more solvents selected from methanol, ethanol, propanol and chloroform and concentrated under reduced pressure. The concentrate was partitioned with hexane, ethyl ether, ethyl acetate, butanol and water, and concentrated under reduced pressure.

The fraction concentrate is purified by silica gel column chromatography, Sep-Pak C18 cartridge and HPLC (High Performance Liquid Chromatography) to obtain an extract with antioxidant activity.

Cucumber herbal extract thus obtained completely removes the toxicity of paraquat, and has strong antioxidant activity due to its excellent scavenging activity by active oxygen species such as hydrogen peroxide, superoxide anion and hydroxyl radical. It can be applied as a therapeutic agent for paraquat antidote, cancer, aging, arteriosclerosis, arthritis, neurodegenerative diseases and the like.

Therefore, the present invention includes an antioxidant composition or paraquat antidote containing the cucumber herbal extract as an active ingredient.

The composition of the present invention can be administered in various oral and parenteral dosage forms during actual clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used, are used. It is prepared by. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the active ingredient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like. Are mixed to prepare. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

The effective dosage of the cucumber herbal extract may vary depending on the age, physical condition, and weight of the patient, but is generally administered within the range of 1 to 100 mg / kg (weight) per day. In addition, within a daily effective dosage range is divided into once a day or several times a day.

In addition, the health food is a food prepared by adding the cucumber herbal extract to food materials such as beverages, teas, spices, gums, confections, or the like, encapsulated, powdered, suspensions, etc. Means to bring, unlike the general medicine has the advantage that there is no side effect that can occur when taking long-term use of the drug as a raw material.

Therefore, the cucumber herbal extract according to the present invention has various antioxidant functions such as inhibiting DNA damage by hydroxyl radical (· OH) and removing superoxide anion (· O ₂ ⁻ ), and has a function of completely detoxifying paraquat. Since the cucumber herbal extract of the present invention and the composition comprising the same is to prevent various diseases that can be caused by oxygen free radicals, development of functional foods, additives for the oxidation of foods, cosmetics with skin aging prevention function, powdery mildew It can be usefully used for pesticides such as control. In addition, it is possible to develop a therapeutic agent, antidote for paraquat poisoning or contaminated patients, and can be easily cultivated and produced at low cost.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1: Isolation and Paraquat Detoxification Effect of Cucumber Herbal Extract

After the development of the cucumber herb leaf 3, only newly emerged new leaves were collected (1.2 kg fresh wt.), And the filtrate was filtered four times with miracloth and extracted four times with the same amount of methanol. It concentrated under reduced pressure (CCA-1100, EYELA). The methanol concentrate was extracted with hexane (n-hexane), ethyl ether (ethyl ether), ethyl acetate (ethyl acetate), butanol and butanol, and concentrated under reduced pressure. The concentrate of each fraction was prepared so that the concentration was 0.1%, and the solution was prepared to have a paraquat concentration of 100 μM and treated with 3 mL per pot on golden bean cotyledon.

As a result of the bioassay by polar fractions, it was confirmed that the water layer extract had antioxidant activity, and the water layer fractions (water soluble fractions) identified with antioxidant activity were subjected to silica gel column chromatography (400 g, 70 to 230 mesh) to inhibit the antioxidant activity. Indicative fractions were obtained. This active fraction was subjected to preparative HPLC using Sep-Pak C18 cartridge to obtain the same active fraction. The active fraction was subjected to silica gel column chromatography (250 g, 70-230 mesh) to obtain a fraction showing antioxidant activity. This active fraction was diluted with chloroform: methanol: water: acetic acid (55: 36: 8: 1, v / v / v / v) and chloroform: methanol: water (6: 4: 1, v / v / v) solvent and butanol. Methanol: Water (4: 1: 2, v / v / v) was analyzed under solvent conditions and developed with ninhydrine. The bioassay confirmed that this fraction was an antioxidant and was analyzed by HPLC (Waters, 2795) using a Cap Cell Pak NH2 column. DPPH free radical scavenging activity was specified for the peaks obtained through the analysis, and the active material was obtained by extractive having antioxidant activity through preparative HPLC.

In each of the solvent fractions, 0.1% of the water-soluble fractions showed a marked effect of reducing paraquat activity. In Paraquat alone treatment, spots were formed on the drug treatment site after 1 day of treatment, and after 3 days, the entire treated area was deciduous.However, when Paraquat was prepared and treated with an aqueous fraction solution, No symptoms were observed. On the other hand, in the hexane, ethyl ether, ethyl acetate and butanol fraction, the mortality by paraquat was 70, 63.3, 63.3 and 43.3%, respectively [Fig. 1].

Example 2: Reduction effect of electrolyte leakage of cucumber herbal extract

Cucumber herbaceous solvent fraction of Example 1 was dissolved in a small amount of distilled water, diluted with a buffer solution (MES), adjusted to a final concentration of 0.1%, poured into 7 mL in a 5 mm diameter Petri dish, and then the golden soybean cotyledon 7 40 lobe sections were teethed using a mm cork borer. Paraquat raw material was prepared to be 100 times the concentration of the treatment with distilled water, and treated with 70 mL each so that the final treatment concentration was 10 μM. The other procedure was followed by Kenyon et al. (1985). The water soluble fraction showed a marked effect of reducing paraquat activity even at the degree of electrolyte leakage. In the 10 μM paraquat treatment, the electrolyte leakage showed a modest increase up to 4 hours of light irradiation, and then rapidly increased to 186.3 μmho / cm after 12 hours. However, when the water-soluble fractions were treated with 0.1% concentration in the same concentration of paraquat, it gradually increased up to 8 hours, and even after 12 hours of light irradiation, the electrolyte leakage was much lower at 50.3 μmho / cm [FIG. 2].

Example 3: Inhibitory Effect of Cucumber Herbal Extract on Malondialdehyde Production

The same method as the electrolyte leakage test was carried out to modify and supplement the methods of Buge (Buege, 1978) and Slater (Slater, 1984), and then to determine the amount of malonaldehyde (MDA) produced using thiobarbituric acid (TBA). Measured. At this time, the treatment concentration of paraquat was 10, 3.3, 1 μM, and the aqueous fraction concentration was 0.1%. Malondialdehyde production in golden soybean cotyledon at paraquat 10, 3.3 and 1 μM concentrations was 24.5, 19.4 and 7.7 nmol / 40 discs, respectively. When 0.1% aqueous fractions were treated at each concentration of these paraquats, the malondialdehyde production was 16.3, 11.1 and 3 nmol / 40 discs, respectively, which significantly reduced the degree of lipid peroxidation (FIG. 3).

Example 4: 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of cucumber herbal extract

1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured using the Blois (1958) method. 990 μl of 100 μM DPPH solution prepared with ethanol was added to 1 ml cuvette, and then ascorbic acid, a-tocopherol and resveratrol prepared at 100 times the desired final concentration. 10 μl each of the cucumber herb water-soluble fractions was stirred with a stirrer and reacted at 20 ° C. for 30 minutes, and the absorbance was measured at 517 nm. 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity was expressed as a decrease in absorbance for no treatment. At this time, alpha-tocopherol, a fat-soluble antioxidant, was used as a water-soluble trolox. Existing antioxidants, ascorbic acid, alphatocopherol and resveratrol, showed concentration-dependent 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Cucumber herbal water-soluble fractions also showed similar trends as 1,1-antioxidant. Diphenyl-2-picrylhydrasil showed free radical scavenging activity. Ascorbic acid, a water-soluble antioxidant, showed a radical scavenging activity of 90.4% at 50 μM concentration, and alphatocopherol showed a scavenging activity of 90.2% at 100 μM concentration. Resveratrol required a relatively high concentration compared to these two antioxidants and showed a scavenging capacity of 78.1% at 250 mM [Fig. 4].

Example 5: Hydrogen Peroxide Scavenging Activity of Cucumber Herbal Extract

Hydrogen peroxide scavenging activity was measured using phenol red and Horseradish peroxidase (HRPase) with reference to the method of Rinkus and Taylor. Phenolic red (phenol red, sodium salt) was prepared in 0.2 M potassium phosphate buffer (pH 6.2) to a final concentration of 7.5 mM, and horseradish peroxidase (HRPase) was added in 0.2 M potassium phosphate buffer. The final concentration was prepared to be 0.5 U / ㎖, where phenol red: horseradish peroxidase was used in a mixture of 1: 2 ratio. Each concentration sample was treated with 300 mM hydrogen peroxide (H ₂ O ₂ ) and allowed to stand for 20 minutes on 30 ℃ growth. This reaction was treated with a phenol red / horseradishperoxidase (phenol red / HRPase) solution and left to stand for 10 minutes after shaking using a stirrer and stored in an ice bath until measurement. Absorbance was measured at 610 nm using a spectrophotometer. The scavenging effect of hydrogen peroxide on hydrogen peroxide was not high. The scavenging capacity of hydrogen peroxide at concentrations of 0.3, 0.6, 1.25, 2.5 and 5 mg / ml was 0.2, 2.2, 5.3, 10.0 and 21.3%, respectively, with superoxide anion or hydroxyl radicals. It was shown to be relatively lower than the scavenging effect for [Fig. 5].

Example 6: Superoxide Anion Radical of Cucumber Herbal Extract ₂ ^- ) Scavenging activity

Superoxide anion scavenging activity was measured by xanthine / xanthine oxydase system and nitro blue tetrazolium (NBT) reduction method. The final amount was added to the reaction to 1.5 ml containing 50 mM potassium phosphate buffer (pH 7.8), 0.05 mM xanthine, 0.6 mM nitroblue tetrazolium (NBT), and the concentration samples. Was added. The reaction was initiated by adding 25 mUm- ¹ xanthine oxidase dissolved in 50 mM potassium phosphate buffer. The reaction was reacted at 25 ° C. for 20 minutes. In this case, a blank of the spectrophotometer was used to insert 50 mM potassium phosphate buffer solution instead of xanthine oxidase, and the control group measured absorbance at 560 nm using a spectrophotometer instead of ascorbate and distilled water in which the sample was dissolved. It was. The scavenging capacity for superoxide anions at concentrations of 0.3, 0.6, 1.25, 2.5 and 5 mg / ml of cucumber herb water-soluble fractions was 2.0, 4.2, 23.2, 43.9 and 71.0%, respectively [Fig. 6].

Example 7: hydroxyl radical (.OH) scavenging activity of cucumber herbal extract

Hydroxyl radical scavenging activity was measured by referring to the deoxyribose method of Elizabeth and Rao (1990). 20 mM potassium phosphate-potassium hydroxide buffer (H ₂ PO ₄ -KOH, pH 7.4), 3 mM 2-deoxyribose to a final volume of 3.5 ml in an eppendorf tube. , 0.1 mM EDTA, 0.05 mM iron chloride (FeCl ₂ ) and samples were treated by concentration, and the reaction was initiated with addition of 3 mM hydrogen peroxide (H ₂ O ₂ ). The reaction mixture was shaken for 30 seconds, stored at 37 ° C. for 1 hour, and the reaction was terminated by adding 0.5% of 2.8% trichloroacetic acid (TCA), followed by 1% TBA dissolved in 50 mM sodium hydroxide (NaOH). 0.5 ml of a reagent (thiobarbituric acid) was added, and the resultant was transferred to a water bath and boiled at 100 ° C. for 10 minutes, and then absorbance was measured at 532 nm using a spectrophotometer. Cucumber herb water-soluble fraction showed a hydroxyl radical scavenging effect in a concentration dependent manner. Hydroxyl radical scavenging ability was 4.8, 11.9, 40.5 and 67.5% when cucumber herb water soluble fraction concentrations were 0.6, 1.25, 2.5 and 5 mg / ml. 7].

Example 8 Inhibition of Xanthine Oxidase Activity of Cucumber Herbal Extract

The inhibitory effect of xanthine oxidase activity was carried out according to the method of Huh et al. (2001). A xanthine solution (100 mM) and xanthine oxidase (EC 1.2.3.2, 0.04 units) prepared in a 1 ml cuvette containing 700 ml of 0.1 M phosphate buffer (pH 7.8) at 10 times the final concentration. And 100 ml of each of the cucumber herb water-soluble fractions were incubated at room temperature for 5 minutes, and the degree of uric acid production was measured at 295 nm. The inhibitory effect of xanthine oxidase activity was measured from the comparison of the absorbance change in the untreated solution with the change in absorbance in the sample treatment group. 8].

Example 9: Toxicity Test

Toxicity experiment was performed on the cucumber herbal extract of Example 1 as follows. Cucumber herb extract was dissolved in dimethylsulfoxide (DMSO), diluted with water, and then administered to each mouse (10 mice per group) 10 g / kg and observed for 7 days, but no rats died.

Preparation Example 1 Preparation of Powder

10 g of active ingredients

50 g of corn starch

40 g of carboxy cellulose

Total amount 100 g

A powder was prepared by crushing and mixing the ingredients listed above. 100 mg of powder was added to the 5 times hard capsule to prepare a capsule.

Preparation Example 2 Preparation of Tablet

10 g of active ingredients

70 g of lactose

15 g of crystalline cellulose

5 g of magnesium stearate

Total amount 100 g

The tablets were prepared by direct tableting method after mixing the ingredients listed above finely. The total amount of each tablet is 100 mg, of which the active ingredient content is 10 mg.

Preparation Example 3 Preparation of Injection

10 mg of active ingredient

Sodium chloride 600 mg

Ascorbic Acid 100 mg

Water for injection

Total amount 100 ml

Injections were prepared in the same composition as above. This solution was placed in an ampoule and heat sterilized at 120 ° C. for 30 minutes.

As described above, the cucumber herbal extract according to the present invention has various antioxidant functions such as inhibition of DNA damage by hydroxyl radical (.OH) and removal of superoxide anion (· O ₂ ⁻ ), which completely detoxifies paraquat. Cucumber herbal extract of the present invention and the composition comprising the same because it has a function to prevent various diseases that can be caused by oxygen free radicals, development of functional foods, additives for the oxidation of foods, preventing skin aging It can be usefully used for cosmetics, pesticides and the like for controlling powdery mildew. In addition, it is possible to develop a therapeutic agent, antidote for paraquat poisoning or contaminated patients, and can be easily cultivated and produced at low cost.

Claims (7)

Cucumis ( Cucumis sativus L.) Antioxidant composition comprising a herbal extract as an active ingredient. Cucumis sativus L. Paraquat antidotes, characterized in that it contains an herbal extract as an active ingredient. Cucumis ( Cucumis sativus L.) Medicine comprising the herbal extract as an active ingredient. Cucumis ( Cucumis sativus L.) A health food comprising herbal extract as an active ingredient. Cucumis ( Cucumis sativus L.) A cosmetic comprising the herbal extract as an active ingredient. Cucumber Cucumis sativus L. Pesticides characterized in that it contains as an active ingredient. 1) extracting the cucumber herb with one or more solvents selected from methanol, ethanol, propanol and chloroform and concentrating under reduced pressure; 2) fractionating the concentrate into hexane, ethyl ether, ethyl acetate, butanol and water, respectively; And 3) Purifying the extract having antioxidant activity by silica gel column chromatography, Sep-Pak C18 cartridge and HPLC (High Performance Liquid Chromatography) on the fraction. Separation method of cucumber herbal extract having an antioxidant activity, characterized in that consisting of.

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