KR101900304B1 - Compositions for Detoxifying Nicotine Containing Green Tea Extract and Black Tea Extract and Method for Preparing the Same - Google Patents

Abstract

The present invention relates to a composition for detoxifying nicotine containing a green tea extract and a black tea extract, and a production method thereof.

Description

TECHNICAL FIELD The present invention relates to a composition for detoxifying nicotine containing a green tea extract and a black tea extract and a method for preparing the same,

The present invention relates to a composition for detoxifying nicotine containing a green tea extract and a black tea extract, and a process for producing the same.

Generally, active oxygen species produced by long-term smoking cause damage to cells by attacking and damaging protein, lipid, nucleic acid, etc. constituting material of cell, thereby causing oxidative damage of these cells and tissues and diseases related to aging and aging, Cancer, duodenal ulcer, ulcerative colitis, bronchitis, cerebrovascular disease, and obstruction. Smoking has also been associated with sleep disturbances, premature menopause, decreased fertility, and impaired motor function. These diseases are caused by 40 kinds of toxic carcinogens in tobacco smoke, among which harmful substances are nicotine, tar and carbon monoxide (Hoffmann, Melikian & Wynder, 1996; Trushine & Hecht, 1998; Witschi, Espiritu, Maronpot, Pinkerton & Jones, 1997; Hecht, 1995; Wynder & Muscat, 1995).

Smoking causes mental and physical dependence, which is caused by the nicotine component in the cigarette. Nicotine is a colorless or light yellow colored poisonous substance that is absorbed into the lungs through smoking and metabolizes in the body at a rapid rate. It acts on the cell membrane of the nerve node and causes an increase in blood pressure, skeletal muscle spasm, And so on. Nicotine also plays an important role in promoting senescence (Damaj, Welch & Martin, 1996).

Nicotine is readily absorbed throughout the body. When you smoke, nicotine reaches the brain after 7 to 19 seconds. This rapid onset of action enhances the smoking behavior. During smoking, nicotine is absorbed into the arteries through the lungs and reaches the brain within a short time. About 70 to 80% of the nicotine inhaled into the body is converted to cotinine, about 10% of the cotinine is excreted in the urine and the remainder is converted to 4-oxo-4 (3-phridyl) butanoic acid (hydroxy acid) (Benoititz, Jacob, Fong and Gupta, 1994).

After smoking, the concentration of nicotine in the arterial blood and brain is rapidly increased, and then decreases over 20 to 30 minutes as the muscles are distributed throughout the body. Nicotine is mainly oxidized to its major metabolite, cotinine, in the kidney, and part of it is excreted into the urine by kidneys as nicotine itself. Therefore, the concentration of nicotine and cotinine in the urine can be a direct measure of smoking and secondhand smoke. In other words, cotinine has a half-life in the body of 17 to 20 hours, which is much longer than 2 hours of nicotine and can be used as a marker of nicotine (Kang, Buseo, 2000).

Several methods have been sought to minimize the risk of smoking, including electronic cigarettes, tobacco pipes, nicotine replacement therapy, and food-grade smoking cessation supplements. This kind of smoking cessation is only to reduce the taste and habit of temporary cigarettes, and it is not a product that can basically quit smoking.

The precise scientific mechanism of the food, which has been reported as a nicotine antidote, has not yet been established. It is also argued that the withdrawal phenomenon can be further increased due to the decrease in the concentration of nicotine, which causes smoking poisoning. In addition, a significant amount of vitamin C is consumed in the body during smoking, while green tea and black tea contain 5-8 times more vitamin C than lemon, and catechin binds nicotine at a very low concentration to form a precipitate (2000). These features complement the existing nicotine antidote problems.

In addition to antioxidant activity, green tea and black tea prevent hypertension and arteriosclerosis, prevent aging by inhibiting lipid peroxidation, prevent obesity by inhibiting neutral lipid production, detoxify toxic heavy metals such as mercury and cadmium, (Kim & Rhee, 1994). It is known that there are many physiological activities such as detoxification by binding.

The difference in composition between green tea and black tea is due to the presence of fermentation. Black tea has a high (multimeric) polyphenol content and green tea has a high (monomeric) polyphenol content. As described above, the green tea and the black tea contain a large amount of catechin and polyphenol. Therefore, it is an object of the present invention to minimize the withdrawal of smoking cessation by increasing the rate of removal of nicotine, which is a toxic substance introduced into the body from cigarette smoke. In addition, green tea or black tea, which is generally consumed with antioxidant function, is considered to be effective in the rejection reaction by smoking cessation.

However, A Study on the Actual Condition of Student Smoking by Measuring Urine Nicotine and Establishment of Student Smoking Rate Monitoring System, Yeungnam University, Final Report of the Ministry of Health and Welfare. 2. Gyeong Yun Joo and Donghee Lee (2000). Promoting Transformation of Nicotine to Cotinine by Green Tea Extract, Journal of the Environmental Agriculture Society of Korea. 19 (2). 147-153. 3. King, Kwak Young-suk, and Ju Jin-hyung (2000). How to abuse drug abuse, Seoul: Shin Il Sang Publishing Co. 4. Benowitz NL, Jacob P, Fong I and Gupta S. Nicotine metabolic profile in man: Comparison of cigarette smoking and transdermal nicotine. J. Pharmacol. Exp. Ther. 1994.268,296-301. 5. Damaj MI, Welch SP & Martin BR. Characterization and modulation of acute tolerance to nicotine in mice. J. Pharmacol. Exp. Ther. 1996.277,454-461. 6. Ghosheh OA, Browne D, Rogers T, De Leon J, Dwoskin LP, Crooks PA. 2000. A simple high performance liquid chromatographic method for the quantification of total cotinine, total 3'-hydroxycotinine and caffeine in the plasma of smokers. J Pharm Biomed Anal 23: 543-549. 7. Hecht SS. Tobacco smoke carcinogens and lung cancer, Natl. Center Inst. 1995.91, 1194-1210. 8. Hoffmann DAA, Melikian EL & Wynder. Scientific challenges in environmental carcinogenesis, Prev. Med. 1996.25,14-22. 9. Kim MJ, Rhee SJ. Effects of Korean green tea, oolong tea and black tea beverage on the removal of cadmium in fat. J Korean Soc Food Nutr. 1994.23: 784-791. 10. Kyerematen GA, Morgan ML, Chattopadhyay B, deBethizy JD, VessllES. 1990. Disposition of nicotine and eight metabolites in smokers and nonsmokers: identification of smokers of two metabolites that are no longer lived than cotinine. Clin Pharmacol Ther 48: 641-651. 11. Trushine N. Hecht. Streoselective metabolism of nicotine and tabacco-specific N-nitrosamines to 4-hydroxy-4- (3-pyridyl) butanoic acid in rats, Chem. Res. Toxicol. 1998.12,164-171. 12. Witschi HI, Espiritu RR, Maronpot KE, Pinkerton AD, Jonse. The carcinogenic potential of the gas phase of environmental tobacco smoke, Carcinogenesis 1997.18,2035-2042. 13. Wynder EL. Muscat. The changing epidemiology of smoking and lung cancer histology, Environ. Health Perspect. 1995.103, 8, 143-148.

Therefore, the present inventors evaluated the ability of nicotine degradation according to the mixing ratio of green tea extract and black tea extract and examined the effect of the mixture of green tea extract and black tea extract on the efficacy of nicotine degrading ability Thus, it was confirmed that nicotine decomposition ability of the mixture of green tea extract and black tea extract was remarkably excellent.

Accordingly, an object of the present invention is to provide a composition for detoxifying nicotine containing a green tea extract and a black tea extract.

Another object of the present invention is to provide a method for preparing a composition for detoxicating nicotine containing a green tea extract and a black tea extract.

Yet another object of the present invention is to provide a nicotine detoxification use of green tea extract and black tea extract.

The present invention relates to a composition for detoxifying nicotine containing a green tea extract and a black tea extract, and a process for producing the same.

The effects of green tea extract and black tea extract on the potency of nicotine degrading ability were investigated by evaluating the ability of nicotine degradation according to the mixing ratio of green tea extract and black tea extract. It was confirmed that the nicotine decomposition ability of the mixture of black tea extract was remarkably excellent.

Hereinafter, the present invention will be described in more detail

One example of the present invention relates to a composition for detoxifying nicotine containing a green tea extract and a black tea extract.

The green tea extract and / or the black tea extract may be in the form of solution, concentrate or powder. The green tea extract and / or the black tea extract may be in the form of a solution, a concentrate or a powder.

The nicotine according to the present invention has the following structural formula (1) and the molecular formula is C ₁₀ H ₁₄ N ₂ . It is a pale yellowish liquid with odor like pyridine. It has a boiling point of 247 ° C (partial decomposition) and a specific gravity of 1.0097. It is highly volatile at room temperature and readily oxidizes to brown when in contact with light or air. The aqueous solution shows weak alkalinity and dissolves in water unlimitedly at 60 ° C or below and 210 ° C or above.

[Chemical Formula 1]

Nicotine is synthesized in the roots of tobacco, and as plants grow in the same way as inorganic salts, they accumulate in the leaves. In leaf, it exists as malic acid and citric acid, and 0.5 to 0.8% or more of dry weight is contained in leaves. Pyridine ring and pyrrole ring. The biosynthetic mechanism is not yet known, but it is theorized that the pyrrole ring is derived from ornithine.

Nicotine is a kind of alkaloid, a compound produced by twin cypress plants. Nicotine is found mainly in tomatoes, potatoes and leaves of Solanaceae plants, especially in the leaves of tobacco. However, the place to be synthesized is roots, which are synthesized in roots and stored in leaves. Nicotine has also been used as an insecticide in the past because of its neurotoxicity to insects. Low levels of nicotine are arousing to humans and other mammals, but higher concentrations can lead to greater toxicity. Humans absorb nicotine mainly through tobacco activities and feel an arousal effect, and nicotine addiction is regarded as one of the most serious addicts to humans.

Also, nicotine-like substances such as noricotine and anabasin are also contained in tobacco, and nicotine is also contained in plants belonging to the genus Hanshako, Ungasia, and the locust bean. Nicotine has strong toxicity, which excites or paralyzes the central nervous and peripheral nerves, contracts the intestines and blood vessels, and promotes the rise in blood pressure. Therefore, acute poisoning by this and the occurrence of chronic poisoning caused by nicotine absorption by smoking are problematic. Nicotine sulphate may be used as an agricultural insecticide (nicotine) and also as a contact agent or fumigant.

The biggest problem with nicotine is dependency. In general, the success rate of smokers quitting smoking is considerably lower, with nicotine dependence being the most likely cause. The major cause of nicotine dependence is known to be the reduced sensitivity of nicotinic acetylcholine receptors and consequently the reduction of dopamine secretion. These nicotine dependence experiments are well underway in rats and gorillas, and it is known that there is actually a dependency effect.

Although nicotine gum or patches are used as a solution to nicotine dependence, the effectiveness of such a method is still controversial. Nicotine is not only a symptom of this addiction but also a substance that is toxic by itself. If you take it directly rather than smoking it can lead to the death of a man by the amount of nicotine contained in two cigarettes. Acute nicotine poisoning can cause vomiting, pulsation, and a rise in breathing. In severe cases, symptoms such as cramping, stunning, and respiratory paralysis occur. Although smoking is known to have a great influence on cancer, nicotine itself does not play an important role in the cancer induction because the carcinogenic effect is not well known.

The green tea according to the present invention keeps green because it stops fermentation by stopping the activity of polyphenol oxidase through high heat to the green leaf or steamed steam. Transparent blue light The tea color and fresh taste are excellent article. In China, more than 70% of the total production of tea is green tea, and in Korea, green tea is being produced mainly in Boseong, Hadong and Jeju Island. The characteristic of green tea is green tea (green tea, green tea, green tea). It has a fresh and elegant fragrance for a long time and the taste is very neat. The fragrance that can be felt in green tea is the scent of the young leaf, the scent of the scent of the scent of the car on the surface of the car, the scent of the night, the scent of flowers, the scent of flowers, and the scent of scent.

The representative types of green tea include Korea's right-wing and trio, Okocha in Japan, West Lake Jungjeong in China, Huangshan Mongbong, Anguil White Tea, and Silver City Jade. Green tea has high content of polyphenol and vitamin C, which is very beneficial for diseases such as hypertension, diabetes, obesity and arteriosclerosis.

The green tea may be one or more selected from the group consisting of leaves, stems, and roots of green tea, and may be, for example, a leaf of green tea.

The green tea extract may be a crude extract obtained by extracting green tea with at least one solvent selected from the group consisting of water and straight chain or branched alcohols having 1 to 4 carbon atoms.

(V / v), more than 10% to less than 100% (v / v), more than 20% to less than 100% (v / v) in the case of using a mixture of water and alcohol in a solvent used for preparing a crude extract of green tea (V / v), less than 100% (v / v), less than 40% to less than 100% (v / Or a branched or unbranched alcohol aqueous solution of 1 to 4 carbon atoms, which is less than 100% (v / v) or less than 70% and less than 100% (v / v)

The alcohol aqueous solution may be at least one selected from the group consisting of an aqueous methanol solution, an aqueous ethanol solution, an aqueous propanol solution, and an aqueous butanol solution.

The green tea extract according to the present invention may be a solvent fraction obtained by fractionating the solvent crude extract with an additional solvent. For example, at least one solvent selected from the group consisting of ethyl ether, ethyl acetate, and butanol is used as the solvent crude extract Solvent fraction.

For example, when the green tea extract is extracted with at least one solvent selected from the group consisting of water and a straight chain or branched alcohol having 1 to 4 carbon atoms, at least one solvent selected from the group consisting of ethyl ether, ethyl acetate, and butanol May be a solvent fraction using a solvent.

The content of the green tea extract as an active ingredient in the composition according to the present invention can be appropriately controlled depending on the mode and purpose of use, the condition of the patient, the type of symptoms and the severity, and is 0.001 to 99.9% by weight, or 0.1 to 99.9% %, Preferably 0.1 to 50 wt%, or 0.1 to 40 wt%.

The black tea according to the present invention is a fermentation difference after progress of fermentation by microorganisms. After the tea is processed first, it is processed again (the method of accumulating the primary processed tea leaves) and the bad process (the method of spraying water to the primary processed tea leaves and increasing the temperature and humidity of the tea leaves itself) So that microorganisms are generated in the car. Black tea is mainly produced in the southwestern regions of China such as Honam, Hubei, Sichuan, Yunnan, and Guangxi. The color of the leaves is black with black or dark brown. The aroma and flavor of black tea is mild and soft and deepens as it grows older. It has a dark red color such as dark red. Black cars are pedestrians who have not oppressed and oppressed cars. Put them in a basket or wrap them in paper, wrap them in bamboo leaves, pack and store them. It is well-known for its six-wheeled cars, four-wheeled cars, The longer you are, the more gentle and tranquil you will be able to drink late at night and the weak and the young children.

The black tea may be one or more selected from the group consisting of leaves, stems and roots of green tea. For example, heat is applied to the leaves of green tea to inactivate enzymes in the tea, and then the microorganisms Means fermented and aged tea, and is distinguished from pre-fermented tea using internal enzymes of tea leaves.

Specifically, the black tea used in the present invention may be contained in the form of an extract, or may be contained as a pulverized product of the black tea itself, or as a dry product of a black tea, but is not limited thereto.

The black tea extract may be a crude extract obtained by extracting a black tea with at least one solvent selected from the group consisting of water and linear or branched alcohols having 1 to 4 carbon atoms.

In the case of using a mixture of water and alcohol in a solvent used for preparing a crude extract of black tea, it is preferable to use 1% to 100% (v / v), 10% to 100% (v / (V / v), less than 100% (v / v), less than 40% to less than 100% (v / Or a branched or unbranched alcohol aqueous solution having 1 to 4 carbon atoms, which is less than 100% (v / v) or less than 70% and less than 100% (v / v)

The alcohol aqueous solution may be at least one selected from the group consisting of an aqueous methanol solution, an aqueous ethanol solution, an aqueous propanol solution, and an aqueous butanol solution.

The black tea extract according to the present invention may be a solvent fraction obtained by fractionating the solvent crude extract with an additional solvent. For example, at least one solvent selected from the group consisting of ethyl ether, ethyl acetate and butanol is used as the solvent crude extract Solvent fraction.

For example, the solvent crude extract obtained by extracting the black tea with at least one solvent selected from the group consisting of water and a straight chain or branched alcohol having 1 to 4 carbon atoms is mixed with at least one selected from the group consisting of ethyl ether, ethyl acetate, and butanol May be a solvent fraction using a solvent.

The content of the black tea extract as an active ingredient in the composition according to the present invention can be appropriately adjusted depending on the mode and purpose of use, patient condition, symptom type, and hardness, and is 0.001 to 99.9% by weight, or 0.1 to 99.9% %, Preferably 0.1 to 50 wt%, or 0.1 to 40 wt%.

The content of the green tea extract and the black tea extract in the composition according to the present invention may be 1 to 99, 1 to 1, 99 to 1.

The composition according to the present invention may be a pharmaceutical composition, a food or a pharmaceutical composition.

When the composition according to the present invention is a pharmaceutical composition, it can be administered to mammals including human in various routes. The mode of administration may be any mode conventionally used and may be administered, for example, by oral, skin, intravenous, intramuscular, subcutaneous, and the like routes, preferably orally. The composition of the present invention can be administered orally or parenterally in the form of powders, granules, tablets, liquids, pills, flakes, gels, bars, jellies, capsules, ointments, suspensions, emulsions, syrups, Parenteral formulations, suppositories, and parenteral formulations in the form of sterile injectable solutions.

The pharmaceutical composition of the present invention may contain pharmaceutically acceptable and physiologically acceptable carriers, excipients, and diluents in addition to the above-mentioned mixed extracts.

Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition of the present invention include dextrin, crystalline cellulose, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, Gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used can be used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose sucrose), lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used.

For oral administration, suspensions, solutions, emulsions, syrups, ointments and the like may be used. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, have.

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, transdermal preparations and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like.

As the suppository preparation, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

In an embodiment where the pharmaceutical composition of the present invention is applied to humans, the pharmaceutical composition of the present invention may be administered alone, but is generally administered in a pharmacological manner selected in consideration of the mode of administration and standard phamaceutical practice. May be administered in admixture with a carrier.

For example, the pharmaceutical composition for detoxifying nicotine containing the green tea extract and the black tea extract of the present invention may be in the form of tablets containing starch or lactose, in the form of capsules containing the active ingredient alone or in an excipient, Oral or sublingual, in the form of an elixir or a suspension containing the active compound, e.g.

Such liquid preparations may contain suspending agents (e. G., A mixture of a semisynthetic glyceride such as methylcellulose, witepsol or apricot kernel oil and a PEG-6 ester or a mixture of PEG-8 and caprylic / Glyceride mixture such as a mixture of glycerides).

The dose of the pharmaceutical composition for detoxifying nicotine containing the green tea extract and the black tea extract of the present invention may be varied depending on the age, body weight, sex, dosage form, health condition and disease severity of the patient, And may be administered once or several times a day at a predetermined time interval.

For example, the daily dose may be 0.1 to 500 mg / kg, preferably 0.5 to 300 mg / kg, based on the active ingredient content. The above-mentioned dosage is an average case, and the dose may be high or low depending on individual differences.

If the daily dose of the pharmaceutical composition for detoxicating nicotine containing the green tea extract and the black tea extract of the present invention is less than the above dose, a significant effect can not be obtained, and if it exceeds the above range, it is not economical, There is a possibility that an undesirable side effect may appear. Therefore, it is preferable to set the above range.

In particular, when the composition for detoxicating nicotine containing the green tea extract and the black tea extract of the present invention is administered to a human body, there is no worry about side effects in comparison with other synthetic drugs in view of the general characteristics of natural extracts. In fact, The toxicity test on the composition for detoxifying nicotine containing the extract and the black tea extract proved to have no effect on the living body.

When the composition according to the present invention is a food composition, the food may be various health functional foods, drinks, food additives, medicines, cosmetics and the like

Examples of the health functional foods include, but are not limited to, capsules, tablets, pills, and liquid preparations.

Examples of the above-mentioned general food include, but are not limited to, canned foods, retorted foods, frozen foods, confectionery, bakery products, jam, processed fish products, beverages, beverages, soups, dressings and liquors.

Examples of the food additives described above may be nutritional enhancers, flavor enhancers, flavoring agents, but are not limited thereto.

The content of the green tea extract and / or the black tea extract as an active ingredient contained in the health functional food is not particularly limited depending on the form of the food, the desired use, etc. For example, 0.01 to 15 wt% And the health beverage composition may be added at a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

The health beverage composition of the present invention contains the green tea extract and the black tea extract as an essential ingredient in the indicated ratios, and there is no particular limitation on the liquid ingredient, and various flavors or natural carbohydrates, such as ordinary beverages, ≪ / RTI >

Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like Sugar, and sugar alcohols such as xylitol, sorbitol, and erythritol.

Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above .

The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition for detoxifying nicotine containing the green tea extract and the black tea extract of the present invention can be used as a flavoring agent, a coloring agent and an aging agent (such as cheese, chocolate, etc.) such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors ), Pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention

Another example of the present invention relates to a method for preparing a composition for detoxicating nicotine containing a green tea extract and a black tea extract.

The green tea extract is a crude extract obtained by extracting at least one member selected from the group consisting of leaves, stems and roots of green tea with at least one solvent selected from the group consisting of water and linear or branched alcohols having 1 to 4 carbon atoms .

The black tea extract is a crude extract obtained by extracting at least one member selected from the group consisting of leaves, stems and roots of green tea with at least one solvent selected from the group consisting of water and straight chain or branched alcohols having 1 to 4 carbon atoms .

The production process of the green tea extract according to the present invention will be described in more detail as follows:

The green tea is washed with water to remove stigmata, dried and then subjected to reflux extraction with an extraction solvent of about 5 to 20 times by volume, preferably 7 to 15 times by volume of the weight of the green tea. After extraction, the filtrate is collected by filtration.

The extraction temperature is not particularly limited, but it is preferably 40 to 110 占 폚, preferably 55 to 90 占 폚.

The extraction process may be repeated once or several times. In a preferred example of the present invention, a method of re-extraction after the first extraction may be adopted. In the case of mass production of herbal extracts, The loss is generated due to the high water content, so that the extraction efficiency is lowered only by the first extraction. In addition, the extraction efficiency of each step was verified, and it was found that about 80 to 90% of the total extraction amount was extracted by the second extraction.

In one example of the present invention, when the extraction step is repeated twice, the obtained residue is subjected to reflux extraction again with an extraction solvent, about 5 to 15 times by volume, preferably 8 to 12 times by volume. After extraction, it is filtered and combined with the filtrate obtained previously and concentrated under reduced pressure to prepare green tea extract. The extraction efficiency can be increased by mixing the extraction solution obtained after the second extraction and the filtrate obtained after each extraction, but the extract of the present invention is not limited to the extraction number.

If the amount of the solvent used in the preparation of the green tea extract is too small, stirring becomes difficult and the solubility of the extract lowers and the extraction efficiency becomes poor. When the amount is too large, the amount of the solvent used in the subsequent purification step increases, The use amount of the solvent is preferably within the above-mentioned range.

The thus-obtained filtered green tea extract is used in an amount of about 10 to 30 times, preferably 15 to 25 times, more preferably about 20 times as much as the total amount of the concentrate for controlling the content of the remaining lower alcohol, The mixture is concentrated by azeotropy with water in a weight ratio of 1 to 5 times, preferably 2 to 3 times, and the same amount of water is added again to homogeneously suspend it, followed by lyophilization to prepare a powdery green tea extract

The production process of the black tea extract according to the present invention will be described in more detail as follows:

The black tea is washed with water to remove stigmata, dried, and then subjected to reflux extraction with an extraction solvent of about 5 to 20 times by volume, preferably 7 to 15 times by volume of the weight of the green tea. After extraction, the filtrate is collected by filtration.

The extraction temperature is not particularly limited, but it is preferably 40 to 110 占 폚, preferably 55 to 90 占 폚.

The extraction process may be repeated once or several times. In a preferred example of the present invention, a method of re-extraction after the first extraction may be adopted. In the case of mass production of herbal extracts, The loss is generated due to the high water content, so that the extraction efficiency is lowered only by the first extraction. In addition, the extraction efficiency of each step was verified, and it was found that about 80 to 90% of the total extraction amount was extracted by the second extraction.

In one example of the present invention, when the extraction step is repeated twice, the obtained residue is subjected to reflux extraction again with an extraction solvent, about 5 to 15 times by volume, preferably 8 to 12 times by volume. After extraction, the mixture is filtered and combined with the previously obtained filtrate, followed by concentration under reduced pressure to prepare a black tea extract. The extraction efficiency can be increased by mixing the extraction solution obtained after the second extraction and the filtrate obtained after each extraction, but the extract of the present invention is not limited to the extraction number.

If the amount of the solvent used in the preparation of the black tea extract is too small, stirring becomes difficult and the solubility of the extract lowers and the extraction efficiency becomes poor. When the amount is too large, the amount of the solvent used in the subsequent purification step is increased, The use amount of the solvent is preferably within the above-mentioned range.

The filtered black tea extract obtained as described above is used in an amount of about 10 to 30 times, preferably 15 to 25 times, more preferably about 20 times as much as the total amount of the concentrate for controlling the content of the remaining lower alcohol, The mixture is concentrated by azeotropic distillation 1 to 5 times, preferably 2 to 3 times, with water in the weight ratio, and the same amount of water is added again to homogeneously suspend it, followed by lyophilization, whereby the extract can be prepared as a powdery black tea extract.

The extraction method used in the present invention may be any conventionally used method such as, but not limited to, cold-water extraction, hot water extraction, ultrasonic extraction, or reflux-cooling extraction.

Another example of the present invention relates to the use of green tea extract and black tea extract for nicotine detoxification.

The green tea extract and the black tea extract are the same as those described above.

The present invention relates to a composition for detoxifying nicotine containing a green tea extract and a black tea extract and a method for preparing the same, wherein the composition for detoxifying nicotine containing a green tea extract and a black tea extract according to the present invention increases the nicotine resolution in the body when smokers ingest It is expected to be able to do.

1 is a diagram showing a human body test progress chart according to an embodiment of the present invention.
FIG. 2 is a graph showing a difference in cotinine change between groups according to an embodiment of the present invention before and after consumption of a green tea and a black tea mixture. FIG.
FIG. 3 is a graph showing a difference in cotinine change between groups after one week of discontinuation of a mixture of green tea and black tea according to an embodiment of the present invention.
FIG. 4 is a graph showing differences in cotinine changes between groups before, during and after 1 week of ingestion of a mixture of green tea and black tea according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments of compositions for detoxifying nicotine.

1-1. Green tea extract manufacturing

1,000 g of purified water was mixed with 100 g of green tea leaves, followed by heating at 80 캜 for 4 hours. After cooling, the mulch was squeezed out, sifted through a 200 mesh screen, and the residue was removed. Thereafter, the mixture was filtered through a 300 meshes screen to remove minute residues. The filtrate was concentrated under reduced pressure to obtain a concentrated green tea extract. The concentrated green tea extract was lyophilized to obtain green tea extract (17.5 g, 17.5%).

1-2. Manufacture of black tea extract

1,000 g of purified water was mixed with 100 g of black tea, and the mixture was kept at 80 캜 and heated for 4 hours. After cooling, the mulch was squeezed out, sifted through a 200 mesh screen, and the residue was removed. Thereafter, the mixture was filtered through a 300 meshes screen to remove minute residues. The filtrate was concentrated under reduced pressure to obtain a concentrated black tea extract. The concentrated black tea extract was lyophilized to obtain a black tea extract (17.9 g, 17.9%).

1-3. Preparation of mixture of green tea extract and black tea extract

(Green tea extract: black tea extract = 3: 1) were uniformly mixed to obtain a green tea-black tea mixed extract (196.0 g, 98%) of 150 g of green tea extract and 50 g of black tea extract.

In Experimental Example 1 below, the nicotine degrading ability was measured according to the mixing ratio of the extract of Example 1.

[ Experimental Example 1 ]

1-1. How to measure nicotine degradation ability

In order to evaluate the nicotine degradation ability by the mixing ratio of green tea extract and black tea extract, the nicotine resolving ability by direct mixing of each water extract and the nicotine resolving ability by cell culture were measured.

<Nicotine metabolism process>

70 to 80% of the nicotine inhaled into the body is converted to cotinine, 10 to 15% of the converted cotinine is excreted in the urine and the remainder is 4-oxo-4- (3-pyridyl) 4- (3-pyridyl)) butanoic acid. (4-oxo-4- (3-pyridyl)) butanoic acid. That is, 85% of keto acid is 4-hydroxy-4- (3-pyridyl) butanoic acid (4-hydroxy-4- (3-pyridyl) butanoic acid) Four percent of the nicotine that is not metabolized to cotinine is converted to nicotine-1'-N-oxide by monooxygenase containing flavin and is mostly excreted in the urine. Nicotine has a half-life of about 30 minutes and has a half-life of more than 15 hours for cotinine. Conversion of nicotine to the detoxified product The content of nicotine was measured by the content of cotinine, which is a stable decomposition product of nicotine.

For HPLC analysis, 2 mL of dichloromethane and 0.6 mL of 5 M NaOH were added to 3 mL of the sample, mixed for 15 minutes, and then centrifuged at 2,000 x g for 5 minutes. The dichloromethane layer was blown off using nitrogen gas and then analyzed with a solution of 0.5 mL of the mobile phase solution. The mobile phase was adjusted to pH 4.7 by adding 0.1 M sodium phosphate / methanol (v / v, 90/10) with triethylamine. The flow rate was 1 mL / min and the UV wavelength was 254 nm.

1-2. Ability to decompose nicotine by mixture

To evaluate the decomposition ability of cotanine of green tea extract and nicotine of black tea extract, 200 ml of 10 mM nicotine, a mixture of green tea extract and black tea extract (0: 1, 1: 3, 1: (1 ug / mL) were mixed and incubated at 37 ° C for 2 hours, and the amount of cotinine produced was measured by HPLC. The results are shown in Table 1 below. .

Green tea extract (%) 0 25 50 75 100 Black tea extract (%) 100 75 50 25 0 Cotinine content (ng / ml) 40.05 87.09 78.67 123.56 52.87 % vs black tea - 117.45% 96.43% 208.51% 32.01% % vs green tea -24.25% 64.72% 48.80% 133.71% -

As shown in Table 1, the decomposition of nicotine into cotinine by the single treatment of green tea extract and black tea extract was 40.05 ng / ml and 52.87 ng / ml respectively, but when the mixture of green tea extract and black tea extract was treated, The extracts of green tea showed higher values of 117.45, 96.43, 208.81 and 32.01% in the ratio of 1: 3, 1: 1, 3: 1 and 1: The ratio of -24.25, 64.72, 48.80, and 133.71% at 1: 0, 1: 3, 1: 1 and 3: 1, respectively.

2-2. Measurement of nicotine degradation ability by cell culture

Hepatocyte-derived Hep-G2 cell line was used for the measurement of nicotine resolution using cell culture. DMEM medium supplemented with 10% FBS, 1% penicillin / streptomycin was added for 1 week in a 5% CO2 incubator The cultured hepatocytes were cultured in DMEM (Dulbecco's modified eagle medium) supplemented with 1 mM nicotine and 5% FBS for 1 day. Then, samples were added to the cultured cells at a concentration of 1 mg / mL and cultured for 3 hours. After culturing, the culture solution and cells were recovered to quantify cotinine.

Cells were washed 3 to 4 times with phosphate buffered saline (PBS) stored at 4 ° C, and cells were collected using a scraper. The collected cells were suspended in 100 μL of PBS and disrupted using an ultrasonic grinder. The cotinine content was measured for the crushed cells and the culture solution, and the results are shown in Table 2 below.

Green tea water extract (%) 0 25 50 75 100 Black tea extract (%) 100 75 50 25 0 Cotinine content (ng / ml) 25.83 82.37 66.08 99.26 40.59 % vs black tea - 218.89 155.83 284.28 57.14 % vs green tea -36.36 102.93 62.80 144.54 -

As shown in Table 2, in the cell culture experiment using the Hep-G2 cell line, a cotinine production pattern similar to that of the direct mixing method was observed. As in the substrate test (Table 1), the highest nicotine resolving power was shown in the ratio of green tea 3: black tea 1, and the substrate test before the green tea and black tea mixture directly affected the nicotine decomposition. , The hepatocyte detoxification ability of the hepatocytes is enhanced, which means that the hepatocytes promote nicotine decomposition.

In the following Experimental Example 2, the change of the nicotine degradation ability according to the ingestion of the green tea extract and the black tea extract was examined.

[ Experimental Example 2 ]

2-1. Target person selection and characteristics

The subjects of this experiment were selected as subjects who used 10 ~ 20 cigarettes daily for more than 1 year for adults 20 years old or older who intend to voluntarily participate and explain the purpose and method of the study. Sixteen patients were randomly assigned to two groups (n = 8 in the test group and n = 8 in the placebo group). The general characteristics of the subject, the daily smoking amount and the smoking characteristics are shown in Tables 3 and 4 below.

kidney weight age BMI The test group (n = 8) 177.88 + - 4.55 77.13 + - 8.58 36.75 ± 6.36 24.30 ± 1.64 The placebo control group (n = 8) 174.75 ± 9.79 76.63 + - 14.85 33.63 + - 5.18 24.90 ± 2.53

Test group
n (%) Placebo control
n (%) χ² (p) Smoking amount
(splint) 5
10
15
20
25
35 0 (0.0)
2 (25.0)
1 (12.5)
4 (50.0)
0 (0.0)
1 (12.5) 1 (12.5)
2 (25.5)
0 (0.0)
4 (50.0)
1 (12.5)
0 (0.0) 4.000 (0.549) Smoking period
(year) 4
5
8
10
12
14
15
20
21 0 (0.0)
1 (12.5)
1 (12.5)
0 (0.0)
1 (12.5)
0 (0.0)
2 (25.0)
2 (25.0)
1 (12.5) 1 (12.5)
0 (0.0)
0 (0.0)
1 (12.5)
0 (0.0)
2 (25.0)
0 (0.0)
4 (50.0)
0 (0.0) 10.667 (0.221)

The subjects were selected by screening test, and the test mixture, green tea and black tea mixture, was taken at a dose of 1000 mg once a day. As shown in FIG. 1, the second test was performed after the first test, and the third test was performed after one week after the first test. The experiment was carried out with a total of 4 measurements including the screening visit. Experimental product composition (range of calculation of items by experimental items)

Item range Caffeine (%) <15 Epigallocatechin gallate (EGCG) (%) 6-14.6 Polyphenol (%) > 36 Total catechin and theaflavin (%) > 18

2-2. Analysis items and measurement methods

Nicotine is influenced by urine pH when excreted in the urine, but cotinine is almost unaffected by urine volume and pH. In addition, the urine cotinine measurement is often less susceptible to contamination in the atmosphere during sampling, even when compared with the carbon monoxide concentration test used in the evaluation of smoking, and the sensitivity and specificity are high. In this experiment, Respectively. Cotinine was measured in urine samples using an ELISA kit, the first urine sample of the first day of the measurement on the day of measurement, which can be measured in blood, saliva and urine, but noninvasive and easily obtainable.

2-3. Statistical analysis

The mean and standard deviation were calculated by descriptive statistics of the data collected using the SPSS (SPSS 21.0 for window, SPSS Inc., Chicago, IL, USA) statistical program. The two-way ANOVA was used for the interaction effect between pre-ingestion and post-ingestion according to experimental group and control group. Statistical significance was set at p <0.05.

2-4. Results of cotinine change measurements before and 1 week after ingestion

As shown in FIG. 2 and Table 6, the change in cotinine before and after consumption of the green tea and black tea mixture in this human body test was significantly higher (470.46 ± 310.77 ng / ml) than that before consumption (709.47 ± 138.15 ng / ml) and 33.69% (39.01 ± 278.34ng / ml), respectively.

In the control group, there was no difference in the mean change between before and after the product (653.60 ± 169.85 ng / ml) and after (654.59 ± 212.78 ng / ml), and the change tendency of cotinine before and after ingestion of green tea and black tea mixture group and placebo group The interaction effect was confirmed by a significant difference (p = 0.040).

group The dictionary (n = 8) Post (n = 8) difference t (p) F (p) mean ± SD mean ± SD mean ± SD Cotinine
(ng / ml) Test group 709.47 ± 138.15 470.46 ± 310.77 -239.01 + 278.34 2.429 (0.046) 5.110 (0.040) Placebo control 653.60 + - 169.85 654.59 ± 212.78 1.00 ± 111.72 -0.025 (0.981) t (p) 0.722 (0.482) -1.383 (0.188)

2-5. One week after ingestion and one week of discontinuation of cotinine measurement

As shown in FIG. 3 and Table 7, in the human body application test, the change of cotinine for 1 week after 1 week of ingestion of the green tea extract and the black tea extract was observed after 1 week of ingestion of green tea and black tea (470.46 ± 310.77 ng / ml) And brown tea extract increased 31% (212.45 ± 270.34 ng / ml) after stopping the mixture for 1 week (682.91 ± 96.80 ng / ml), but there was no statistically significant difference.

In the placebo group, the mean increase was 657.59 ± 212.78 ng / ml and 707.77 ± 198.09 ng / ml, respectively, but not statistically significant. Therefore, there was no significant interaction effect between experimental group and placebo group after 1 week of discontinuation of mixture of green tea extract and black tea extract.

group One week after ingestion (n = 8) One week after discontinuation (n = 8) difference t (p) F (p) mean ± SD mean ± SD mean ± SD Cotinine
(ng / ml) Test group 470.46 ± 310.77 682.91 + - 96.80 212.45 ± 270.34 -2.223 (0.062) 2.259 (0.155) Control group 654.59 ± 212.78 707.77 ± 198.09 53.17 ± 129.45 -1.162 (0.283) t (p) -1.383 (0.191) -0.319 (0.754)

2-6. Measurements of cotinine change before ingestion, one week after ingestion and one week after ingestion

As can be seen from FIG. 4 and Table 8, the change in cotinine before, 1 week after ingestion and 1 week after ingestion of green tea extract and black tea extract in this human body test showed that the preincubation (709.47 ± 138.15ng / ml), intake (470.46 ± 310.77 ng / ml) and one week after discontinuation (682.91 ± 96.80 ng / ml) were decreased after ingestion compared to before consumption, but increased after 1 week after stopping, (P = 0.044). There was no statistically significant difference between pre- and post-

group Before ingestion (n = 8) One week after ingestion (n = 8) After 1 week of discontinuation (n = 8) F (p) mean ± SD mean ± SD mean ± SD Cotinine
(ng / ml) Test group 709.47 ± 138.15 470.46 ± 310.77 682.91 + - 96.80
5.007 (0.014)
Control group 653.60 + - 169.85 654.59 ± 212.78 707.77 ± 198.09 t (p) 0.722 (0.482) -1.383 (0.188) -0.319 (0.754)

[ Experimental Results ]

The results were as follows: 1) When a mixture of green tea extract and black tea extract was consumed for 1 week, green tea extract and black tea extract were mixed for 1 week and 1 week after stopping the administration of test group (mixture of green tea extract and black tea extract) (P = 0.040), and 2) green tea extract and black tea extract were consumed for 1 week, but there was no significant increase in cotinine after 1 week (p = 0.062) and the likelihood of persistence of efficacy for 1 week after ingestion was confirmed. Therefore, it was confirmed that smokers could increase the nicotine resolution in the body when they take a mixture of green tea extract and black tea extract at a dose of 1000 mg per day.

It is to be understood that the invention is not limited to the form set forth in the foregoing description. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims. It is also to be understood that the invention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention as defined by the appended claims.

Claims (7)

Green tea extract and black tea extract,
Wherein the weight ratio of the green tea extract to the black tea extract is 3: 1 to 1: 3.
delete delete delete delete delete Preparing a green tea extract, preparing a black tea extract;
Mixing the green tea extract and the black tea extract at a weight ratio of 3: 1 to 1: 3;
Wherein the method comprises the steps of:

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