WO2021038904A1 - Manufacturing method for composition with reduced urushiol content, composition manufactured using said manufacturing method, obesity prevention agent, fat accumulation suppression agent, and serum leptin quantity increase suppression agent - Google Patents

Abstract

Provided is a manufacturing method for a lacquer tree-derived composition that can secure an effective component of the lacquer tree while removing urushiol in a simple step. A composition with a reduced urushiol content is manufactured using a step in which a solution including a mixed solvent of water and alcohol and an extracted lacquer tree component within said mixed solvent is condensed to obtain a condensed solution that is one-half of the volume of the solution or less.

Description

A method for producing a urushiol content-reducing composition, a composition produced by the production method, an anti-obesity agent, an adipose accumulation inhibitor, and a blood leptin amount increase inhibitor

The present invention relates to a method for producing a sumac-derived composition capable of reducing the content of urushiol, and a composition useful as an anti-obesity agent, an inhibitor of fat accumulation, and an inhibitor of an increase in blood leptin amount.

Research on the development of health drinks and pharmaceutical compositions using ingredients obtained from plants growing in nature has been actively conducted. Among them, there are studies to extract the components contained in sumac and evaluate the medicinal effect, and as a result of these studies, it has been reported that sumac contains various active ingredients. (Non-Patent Documents 1 to 4).
Here, sumac contains various active ingredients, and is highly expected as a raw material for pharmaceuticals. However, it is also known that urushiol contained in sumac has problems such as inducing strong allergic symptoms such as allergic contact dermatitis (Non-Patent Document 5). Therefore, in order to safely use the active ingredient contained in sumac, it is necessary to extract the active ingredient from sumac and remove urushiol. For this reason, conventionally, the active ingredient is obtained from sumac by carrying out two steps, a step of removing urushiol from sumac and a step of extracting the active ingredient.

However, the method of obtaining the active ingredient of sumac in two steps requires chemicals and equipment in each step, and doubles the labor by the number of steps, which requires cost and time. The issue is that the production efficiency is low. In addition, since there is a loss of components in each process, there is also a problem that the yield of the obtained active ingredient is low.
Therefore, in order to solve the problems of the prior art, the present inventors provide a method for producing a sumac-derived composition capable of removing urushiol in a simple step while securing the active ingredient of sumac. We proceeded with the study for the purpose.

The present inventors have conducted diligent studies in order to solve such problems of the prior art. As a result, it was found that there is a method in which the active ingredient of sumac can be concentrated while significantly removing urushiol by performing a one-step treatment on the solution containing the ingredient of sumac. The present invention has been proposed based on these findings, and specifically has the following configuration.

[1] A step of concentrating a solution containing a mixed solvent of water and alcohol and a component of sumac extracted in the mixed solvent to obtain a concentrated solution having a volume of the solution reduced to half or less. A method for producing a urushiol content-reducing composition.
[2] The production method according to [1], wherein the sumac is a leaf of sumac.
[3] The production method according to [1] or [2], wherein when the alcohol concentration before concentration is a% by volume and the alcohol concentration after concentration is b% by volume, ab is 10% by volume or more. ..
[4] The production method according to any one of [1] to [3], wherein the concentration reduces the alcohol concentration of the solution to less than 50% by volume.
[5] The production method according to any one of [1] to [4], wherein the concentration is carried out under reduced pressure.
[6] The production method according to any one of [1] to [5], wherein the concentration is carried out at 20 to 50 ° C.
[7] The production method according to any one of [1] to [6], wherein urushiol contained in the solution is precipitated by the concentration.
[8] The production method according to any one of [1] to [7], wherein the concentration reduces the concentration of urushiol contained in the solution to less than 1/10.
[9] The production method according to any one of [1] to [8], wherein the urushiol concentration of the concentrated solution is less than 0.1% by weight.
[10] The production method according to any one of [1] to [9], wherein 80% by weight or more of kercitrin contained in the solution before concentration remains in the concentrated solution after concentration.
[11] The production method according to any one of [1] to [10], further comprising a step of drying the concentrated solution into a powder.
[12] A composition produced by the production method according to any one of [1] to [11].
[13] The composition according to [12], which further contains one or more selected from the group consisting of elm, oyster, lychee, litchi and keninjin.
[14] An anti-obesity agent comprising the composition according to [12] or [13].
[15] A fat accumulation inhibitor containing the composition according to [12] or [13].
[16] An inhibitor for increasing the amount of leptin in blood, which comprises the composition according to [12] or [13].

According to the production method of the present invention, urushiol can be removed from the sumac extract in a simple step while reliably leaving the active ingredient of sumac. Therefore, according to the production method of the present invention, a sumac-derived composition having a reduced urushiol content can be efficiently obtained at a low production cost.
Since the composition produced by the production method of the present invention has a reduced urushiol content, it can be safely used as an obesity inhibitor, a fat accumulation inhibitor, and a blood leptin amount increase inhibitor.

It is a graph which shows the change of the solution volume, ethanol concentration, urushiol residual rate and cercitrin residual rate in the process of concentrating lacquer extract. A group of mice fed a high-fat diet (Group I), a group of mice fed a high-fat feed containing 1% by weight of dried urushi leaf extract powder (Group II), and a high content of 2% by weight of dried urushi leaf extract powder. It is a graph which shows the weight change of the mouse group (group III) which fed a fat feed. A group of mice fed a high-fat diet (Group I), a group of mice fed a high-fat feed containing 1% by weight of dry urushi leaf extract powder (Group II), and a high content of 2% by weight of dry urushi leaf extract powder. It is a graph which shows the intraperitoneal fat mass after 56 days feeding of the mouse group (group III) which fed a fat feed. A group of mice fed a high-fat diet (Group I), a group of mice fed a high-fat feed containing 1% by weight of dried urushi leaf extract powder (Group II), and a high content of 2% by weight of dried urushi leaf extract powder. It is a graph which shows the amount of blood leptin immediately before feeding (day 0) and the amount of blood leptin after feeding for 56 days of a group of mice fed with a fat feed (group III).

The present invention will be described in detail below. The description of the constituent elements described below may be based on typical embodiments or specific examples, but the present invention is not limited to such embodiments. In this specification, the numerical range represented by using "-" means a range including the numerical values before and after "-" as the lower limit value and the upper limit value.

<Manufacturing method of urushiol content reduction composition>
In the method for producing a urushiol content-reducing composition of the present invention, a mixed solvent of water and alcohol and a solution containing the components of sumac extracted in the mixed solvent are concentrated to halve the volume of the solution. It is characterized by including a step (concentration step) of obtaining the concentrated solution described below.

The lacquer tree used in the production method of the present invention can be selected from those belonging to the lacquer family, and a plant species belonging to the genus Rhus family is particularly preferable. For example, Rhus verniciflua, Rhus trichocarpa, Rhus ambigua, Rhus javanica, Yamahaze (Rhus sylvestris) can be exemplified, but not limited to these.
The poison ivy is composed of leaves, roots, trunks, branches, etc., and any part may be used for the manufacturing method, but the leaves and the bark and xylem of the trunks and branches are easy to collect, especially the leaves. It is preferable to use a portion. Although the bark and xylem have been actively studied, there are not many research examples on the leaves of sumac, and by reducing the urushiol content by the production method of the present invention, research and development of the active ingredient thereof has greatly progressed. Can be made to.

The leaves of poison ivy can be used in a state where they can be easily extracted. For example, it may be extracted in the form of fragments or powder.
When the leaves are shredded or powdered, the collected leaves may be treated as they are with a cutter, a shredding machine, a colloid mill, etc., but once dried, shredded and powdered. It is preferable to do so. The leaves are generally dried until the water content is less than 10% by weight, preferably less than 5% by weight, more preferably less than 3% by weight. The drying may be natural drying or mechanical drying. In addition, when drying, it is preferable to carry out within 30 minutes from the leaf collection.

The drying temperature of the leaves is not particularly limited. Therefore, rapid heating and drying may be performed using a pressurized drum heating device, electromagnetic waves, or the like. When a pressurized drum heating device is used, it is preferable to heat and dry it in the range of 80 to 140 ° C. The drying time is usually 2 minutes or less, preferably 1 minute or less, and more preferably about 40 to 50 seconds. When an electromagnetic wave such as a microwave oven is used, it can be heated and dried at 600 W for about 20 to 50 seconds, for example. By heating and drying under such conditions, the activity of undesired enzymes contained in the leaf raw material can be suppressed or inactivated, and the decomposition of physiologically active components can be prevented to some extent.

The product that has been rapidly heated and dried may be used as it is, or may be further dried at a low temperature before use. When low temperature drying is performed, it is preferably performed in the range of −5 ° C. to 10 ° C. The low temperature drying can be performed by using a hot air drying device such as a far infrared drying device, a ventilation drying device, an ice temperature drying device, or the like alone or in combination. For example, it can be dried at ice temperature after being dried by far infrared rays. By performing such low-temperature drying, decomposition of the physiologically active component can be prevented.

The leaves can be shredded and pulverized using equipment and tools suitable for the purpose. For example, it can be made into a powder of 50 to 100 μm using a colloidal mill. Such shredding or pulverization may be performed before drying, after high temperature drying, or after low temperature drying.

The leaf portion to be extracted may be not only the leaf portion powdered or fragmented in this way, but also the collected leaf portion itself, a high-temperature dried leaf, a low-temperature dried leaf, or the like. In order to improve the extraction efficiency, it is preferable that the leaves are fragmented to some extent.

The extraction solvent is a mixed solvent of alcohol and water. Examples of the alcohol include methanol, ethanol, propanol, isopropanol, butanol, isobutanol, tert-butanol, pentanol, isopentanol, hexanol, and isohexanol. Methanol, ethanol, and propanol are preferable, and ethanol is preferable. More preferred. Only one of the alcohols may be used, or a combination of alcohols may be used. The alcohol concentration of the extraction solvent is preferably 60% by volume or more, more preferably 65% by volume or more, and further preferably 70% by volume or more. The alcohol concentration of the extraction solvent is preferably 85% by volume or less, more preferably 80% by volume or less, and further preferably 75% by volume or less.

Extraction of the components of sumac is carried out by bringing a mixed solvent of alcohol and water into contact with sumac and eluting the components of sumac into the mixed solvent. As a result, a mixed solvent of alcohol and water and a solution containing the components of sumac extracted in the mixed solvent are obtained. In the following description, this solution is referred to as "lacquer extract". Extraction can be carried out by immersing the sumac in a mixed solvent of alcohol and water and leaving it for a certain period of time. This extraction may be carried out at room temperature or under reflux. Moreover, it is preferable to carry out while stirring the solution. Further, extraction may be performed using an extraction device such as a Soxhlet extractor.
The obtained sumac extract may be subjected to the concentration step as it is, but it is preferable that the insoluble matter is removed by filtration or the like before the concentration step.

In the concentration step of the present invention, the solution of sumac extract prepared as described above is concentrated to obtain a concentrated solution having a volume reduced to less than half of the volume before concentration. "Concentration" in this concentration step means removing a part of the mixed solvent from the mixed solvent of alcohol and water and the solution containing the components of sumac extracted in the mixed solvent (urushi extract). To do.
In this concentration process, urushiol is precipitated and the urushiol content in the solution is reduced. By removing the precipitated urushiol from the concentrated solution by filtration or the like, a composition having a reduced urushiol content can be obtained. Thereby, in the production method of the present invention, the urushiol content of the solution can be significantly reduced while the active ingredient of sumac is surely left in the solution by a simple step of concentrating the solution.

The solution can be concentrated, for example, by evaporating a part of the mixed solvent contained in the solution. In this case, it is preferable to evaporate at least a part of alcohol, and it is preferable that the proportion of alcohol evaporated is larger than the proportion of water evaporated. Here, the ratio of the evaporated portion means the ratio of the volume lost due to evaporation to the volume before concentration. The concentration may be carried out under normal pressure or reduced pressure, but it is preferably carried out under reduced pressure. As a result, the mixed solvent can be evaporated at a low temperature, so that thermal denaturation and thermal decomposition of the physiologically active component can be suppressed. The temperature for concentration varies depending on the pressure conditions, but is preferably 20 to 50 ° C, more preferably 25 to 45 ° C.

In this concentration step, the alcohol concentration of the solution is reduced by concentrating the solution. When the alcohol concentration before concentration is a% by volume and the alcohol concentration after concentration is b% by volume, ab is preferably 10% by volume or more, more preferably 20% by volume or more, and 30% by volume. It is more preferable that the volume is% or more.
Further, it is preferable to reduce the alcohol concentration of the solution to less than 50% by volume, more preferably to less than 45% by volume, and further preferably to less than 40% by volume by concentrating the solution.
Further, by concentrating the solution, the concentration of urushiol contained in the solution is preferably reduced to less than 1/10 of the concentration of urushiol before concentration, more preferably less than 1/20, and 30 It is more preferable to reduce it by less than one-half.

In the present invention, the volume of the concentrated solution obtained in the concentration step is 1/2 or less, may be 1/3 or less, 1/5 or less, 1/7 or less, 1 / of the volume of the solution before concentration. It may be 9 or less. The lower limit of the volume of the concentrated solution is 1/20 from the viewpoint that urushiol can be easily removed by the filtration operation.

The urushiol concentration of the concentrated solution obtained in the concentration step is preferably less than 0.1% by weight, more preferably less than 0.05% by weight, still more preferably less than 0.01% by weight. , More preferably less than 0.005% by weight.
Further, it is preferable that 80% by weight or more of kercitrin contained in the solution before concentration remains in the concentrated solution after concentration, and 85% by weight or more of kercitrin contained in the solution before concentration is concentrated after concentration. It is more preferable that it remains in the solution.
The content of each component contained in the sumac extract or concentrated solution can be measured by high performance liquid chromatography (HPLC). For the specific measurement conditions, the description in the column of Examples can be referred to.

The obtained concentrated solution may be used as it is as a composition, but in order to enhance the effect, it is also preferable to carry out a step of drying the concentrated solution into a powder. The concentrated solution may be dried by natural drying or freeze-drying. Further, after powdering, the concentration may be adjusted by further dissolving in physiological saline or the like. In addition, heat sterilization may be performed before powdering.

<Composition>
The urushiol content-reducing composition (composition of the present invention) produced by the production method of the present invention contains a physiologically active substance (active ingredient) such as quercitrin and polyphenol derived from sumac, and induces allergies. Since the content of urushiol is significantly reduced, it is useful as a pharmaceutical composition and a health drink. Therefore, a prophylactic or therapeutically effective amount of the composition of the present invention can be used to prevent or treat a disease by administering it to mammals including humans.

The composition of the present invention can be used as an anti-obesity agent and an inhibitor of fat accumulation. Therefore, by administering an effective amount of the anti-obesity agent and the anti-fat accumulation inhibitor of the present invention to a living body, the accumulation of body fat such as intraperitoneal fat is suppressed and obesity is effectively prevented. can do.
In addition, the composition of the present invention can be used as an inhibitor for increasing the amount of leptin in blood. Leptin is an adipose tissue-derived hormone that activates leptin receptors in hypothalamic neurons to suppress feeding and increase energy expenditure, while it is known that the higher the degree of obesity, the higher the amount of leptin in the blood. Has been done. Therefore, the amount of leptin in blood is generally used as an index of the degree of obesity, and the lower the amount of leptin in blood, the more obesity is suppressed. It is presumed that when an effective amount of the blood leptin amount increase inhibitor of the present invention is administered to a living body, fat accumulation is suppressed and obesity is prevented, and as a result, the increase in blood leptin amount is suppressed. In addition, the fact that the action of suppressing the increase in the amount of leptin in blood of the present invention does not cause the insufficient action of leptin is that the amount of leptin in blood is not caused by the system to which the urushiol content reducing composition of the present invention is applied in the following examples. It can be confirmed from the fact that weight gain and fat accumulation are also suppressed at the same time as the increase in leptin is suppressed.

The composition of the present invention can be used in various forms depending on its intended use.
For example, when the composition of the present invention is used as a pharmaceutical product, various dosage forms can be selected depending on the route of administration thereof. The compositions of the present invention can be administered orally or parenterally. For example, rectal administration, nasal administration, buccal administration, sublingual administration, intravaginal administration, intramuscular administration, subcutaneous administration, and intravenous administration can be performed. Among them, the composition of the present invention is preferably orally administered, subcutaneously or transdermally administered, and is particularly preferably orally administered.

Examples of preparations suitable for oral administration include tablets, capsules, powders, fine granules, granules, liquids, and syrups, and preparations suitable for parenteral administration include injections, infusions, and suppositories. , Inhalants, transdermal absorbents, transmucosal absorbents, patches and the like. The injection may be used for any of intravenous injection, intramuscular injection, subcutaneous injection, infusion and the like. The composition of the present invention is particularly preferably any of oral preparations, injections, and patches.

Pharmacologically and pharmaceutically acceptable additives can be added to the composition of the present invention, if necessary. For example, excipients, disintegrants or disintegrants, binders, lubricants, surfactants, coatings, dyes, diluents, bases, solubilizers or solubilizers, isotonic agents, pH adjusters. , Stabilizers, propellants, adhesives, wetting agents and the like can be used. In addition, other physiologically active ingredients (for example, one or more selected from the group consisting of elm, wolfberry, wolfberry (wolfberry), litchi and carrot (carrot)) may be added. By using these additives in appropriate combinations, the composition of the present invention can be provided with various additional functions.

Specific examples of the above excipients include starch, cornstarch, sucrose, lactose, mannitol, crystalline cellulose, carboxymethyl cellulose, gum arabic, and inorganic salts.

Examples of the disintegrant or disintegrant include wheat starch, rice starch, corn starch, potato starch, partially pregelatinized starch, hydroxypropyl starch, sodium carboxymethyl starch, tragant, crystalline cellulose, methyl cellulose, and low-substituted hydroxypropyl cellulose. Specific examples include carmellose, carmellose calcium, carmellose sodium, and croscarmellose sodium.

Examples of the binder include agar, gelatin, wheat starch, rice starch, corn starch, potato starch, dextrin, pregelatinized starch, partially pregelatinized starch, hydroxypropyl starch, crystalline cellulose, crystalline cellulose / carmellose sodium, methyl cellulose, and hydroxy. Propyl cellulose, low substitution hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose phthalate, hydroxypropyl methyl cellulose acetate succinate, carmellose sodium, ethyl cellulose, carboxymethyl ethyl cellulose, hydroxyethyl cellulose, purulan, polyvinylpyrrolidone, aminoalkyl methacrylate copolymer, methacryl Specific examples include acid copolymers, polyvinyl acetal diethylaminoacetate, polyvinyl alcohol, gum arabic, powdered arabic rubber, white cellac, tragant, purified sucrose, and macrogol.

Examples of the lubricant include wheat starch, rice starch, corn starch, sucrose fatty acid ester, stearic acid, calcium stearate, magnesium stearate, hydrous silicon dioxide, light anhydrous silicic acid, synthetic aluminum silicate, and dry aluminum hydroxide gel. , Talk, magnesium aluminometasilicate, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, braces, hydrogenated oil, polyethylene glycol are specific examples.

Examples of the surfactant include sucrose fatty acid ester, soybean lecithin, polyoxyl stearate, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene glycol, sorbitan sesquioleate, sorbitan trioleate, sorbitan monostearate, and mono. Specific examples include glycerin stearate, sorbitan monopalmitate, sorbitan monolaurate, polysorbate, sodium lauryl sulfate, and lauromacrogol.

Further, the composition of the present invention can be designed so that the active ingredient is slowly released as needed. It can also be designed so that the active ingredient is released intensively where it is needed in the body. Such sustained-release preparations and drug delivery systems can be designed and manufactured according to methods well known in the pharmaceutical industry.

Further, an organic or inorganic carrier can be used in the composition of the present invention. Examples of such carriers include lactose, starch, vegetable and animal fats and fats and oils.

The administration target can be mammals other than humans such as dogs and humans. The dose of the composition of the present invention is appropriately determined according to various conditions such as the purpose of treatment or prevention, the sex, body weight, age of the patient, the type and degree of disease, the dosage form, the route of administration, and the number of administrations. For example, in the case of oral administration, 0.1 μg to 100 mg (dry weight of sumac leaf extract) / kg body weight / day can be administered once to several times a day, but the dose is in this range. It is not limited to.

The composition of the present invention does not necessarily have to be in the form of a pharmaceutical product. For example, the composition of the present invention can be safely and effectively used as a functional food or a functional beverage by including it in various foods and beverages. In particular, it can be effectively used as a food or beverage labeled to have an anticancer effect.
Specifically, it can be used for black tea, soft drinks, juices, candy, starchy foods, various processed foods, etc., but the usage form is not limited to these specific examples. Further, if necessary, a gelling agent or the like may be added to improve the texture.

The features of the present invention will be described in more detail below with reference to Examples and Comparative Examples. The materials, amounts used, ratios, treatment contents, treatment procedures, etc. shown in the following examples can be appropriately changed as long as they do not deviate from the gist of the present invention. Therefore, the scope of the present invention should not be construed as limiting by the specific examples shown below. In the following description, "%" representing the mixing ratio of the mixed solvent means "volume%".

The component analysis, antioxidant activity and α-glucosidase inhibitory activity of the composition prepared in this example were measured by the following methods.
(A) Component analysis by high performance liquid chromatography (HPLC) Quercetin, quercetin, sulfretin, fisetin, fustin and the like were component-analyzed by an HPLC system (Agilent Technologies, Inc .: Agilent 1100 series). Specifically, a linear gradient elution (95:) using a reverse phase column (manufactured by Tosoh Corporation: TSKgel ODS-80Ts (inner diameter 4.6 mm × length 250 mm)) and using a 10 mM phosphoric acid-acetonitrile mixed solvent as a mobile phase. 5 → 5: 95, 60 minutes) was carried out to detect each component and measure the content.
For urushiol, 90% acetonitrile was eluted as a mobile phase using an HPLC system (Shimadzu: LC6A system) and a reverse phase column (Nomura Chemical: Develosil ODS-5 (inner diameter 10 mm x length 250 mm)). Then, detection was performed at a detection wavelength of 272 nm, and the content was measured.
(B) Measurement of total polyphenol content The total polyphenol content was measured using the Folin-Ciocalt method.
(C) Measurement of Alcohol Concentration The alcohol concentration was measured using an alcohol concentration meter (manufactured by AS ONE Corporation: DAAL-25).
(D) Measurement of antioxidant activity Measurement of DPPH radical scavenging activity:
In a 96-well microplate, 60 μL of the sample solution for measurement, 60 μL of 0.2 M MES (2-morpholinoatherulphonic acid) buffer (pH 6.0), 60 μL of 20% ethanol solution, and 400 μM DPPH (2,2) dissolved in ethanol. 60 μL of a solution of −Diphenyl-1-picrylhydrazyl) was added, and the reaction was carried out at room temperature for exactly 20 minutes. Absorbance at 520 nm was measured using a microplate reader. A regression line was prepared using Trolox, and the amount was calculated as the amount of Trolox (μmol Trolox / g) corresponding to the amount of sample added (μL / assay).
Measurement of Oxygen Radical Absorption Capacity (ORAC):
To a 96-well microplate, 25 μL of the sample solution for measurement and 150 μL of the Fluorescein solution (dissolved in 75 mM phosphate buffer (pH 7.4)) were added, and the mixture was incubated at 37 ° C. for 30 minutes. Immediately after adding 25 μL of AAPH (2,2'-azobis (2-methylpropionamide) dihydrochloride) solution and stirring with shaking, the fluorescence intensity was measured using a plate reader maintained at 37 ° C. (Em: 485 nm, Ex: 528 nm). .. Then, the change with time of the fluorescence intensity was measured for 60 minutes at 5-minute intervals. The area under the curve of the graph recording the fluorescence intensity was calculated, and the ORAC value was displayed as the amount equivalent to Trolox (μmol Trolox / g) per 1 g of sample weight.
(E) Measurement of α-glucosidase inhibitory activity To a 96-well microplate, add 10 μL of a sample solution for measurement and 40 μL of yeast-derived α-glucosidase (0.1 mU / mL) dissolved in 67 mM phosphate buffer (pH 6.8). , Preheated at 37 ° C. for 15 minutes. Subsequently, 50 μL of p-nitrophenyl α-D-glucopyranoside (1 mM) dissolved in 67 mM phosphate buffer (pH 6.8) was added to initiate the reaction. After reacting at 37 ° C. for exactly 30 minutes, 100 μL of 0.5 M sodium carbonate solution was added and mixed to stop the reaction. Absorbance at 400 nm was measured using a microplate reader. The α-glucosidase inhibition rate was calculated using the following formula.
Inhibition rate (%) = {(CS) / (CB)} x 100
C: Absorbance at 400 nm of the control S: Absorbance at 400 nm of the sample B: Absorbance at 400 nm of the blank Note that the final concentration of the sample during the enzymatic reaction when the inhibition rate shows 50% is defined as the IC50 value, which is α. -Used as a measure of glucosidase inhibitory activity.

(Example 1) Production and evaluation of dry powder of sumac leaf extract (composition for reducing urushiol content) 70% ethanol / 30 with respect to 800 g of dry powder of sumac leaves (Rhus verniciflua leaf) collected in Wonju, South Korea. % Water mixed solvent (10 times amount, 8 L) was added, and the mixture was stirred at room temperature (25 ° C.) for 16 hours to obtain a sumac leaf extract. The sumac leaf extract obtained here corresponds to a solution containing a 70% ethanol / 30% water mixed solvent and the components of sumac extracted in the mixed solvent. The obtained sumac leaf extract was suction-filtered using a Buchner funnel equipped with a filter paper (No. 131). The filtrate was concentrated under reduced pressure to about 1/10 volume (about 800 mL) in a hot water bath at 40 ° C. using a rotary evaporator. The concentration process produced a black precipitate containing urushiol. Subsequently, in order to separate the concentrated solution and the black precipitate containing urushiol, suction filtration of the concentrate was performed again using a Büchner funnel equipped with filter paper (No. 131), and the filtrate (concentrated solution) was recovered. ..
When the components of the obtained concentrated solution were analyzed, urushiol was not detected, and it was confirmed that it contained quercitrin and polyphenols.
Then, the concentrated solution is sterilized by heating at 90 ° C. for 5 minutes, allowed to cool sufficiently, pre-frozen at -80 ° C., and further freeze-dried in a freeze-dryer for about 6 days to dry the sumac leaf extract. A powder (composition 1) was obtained.

(A) Component analysis The dry powder of sumac leaf extract obtained in Example 1 was subjected to component analysis by high performance liquid chromatography (HPLC). As a result, quercetin was found to be contained in the bark and xylem. Quercetin, sulfretin, fisetin, fustin, etc. were not detected.

(B) Measurement of changes in the contents of urushiol and quercitrin in the concentration process In order to investigate changes in the contents of urushiol and quercitrin in the concentration process, a 70% ethanol / 30% water mixed solvent was used under the same conditions as in Example 1. The lacquer leaf extract was prepared, and the lacquer leaf extract (900 mL) was concentrated under reduced pressure to about 1/10 volume (100 mL). In this concentration process, a part of the solution (solution sample) is taken step by step, the ethanol concentration, urushiol content and cercitrin content of the solution sample are measured, and from the measurement results, when the solution sample is taken. The ethanol concentration of the solution, the residual rate of urushiol, and the residual rate of quercitrin were determined. The result is shown in FIG. In FIG. 1, the horizontal axis shows the volume of the solution when the solution sample was taken, the left vertical axis shows the ethanol concentration of the solution, and the right vertical axis shows the urushiol residual rate and the cercitrin residual rate of the solution. .. Here, the residual ratio means the ratio of the content at each time point when the content before concentration is 100% by weight.
As shown in FIG. 1, the urushiol content decreased remarkably as the concentration progressed, while the useful ingredient quercitrin was hardly lost in the concentration process and maintained the initial content. From this, it was found that the urushiol content can be selectively reduced by concentrating the sumac extract with a mixed solvent of ethanol and water.

(Comparative Example 1) Preparation and comparative evaluation of unconcentrated sumac leaf extract Add 10 times (8 L) of ethanol / water mixed solvent to 800 g of dry sumac leaf powder and stir at room temperature (25 ° C) for 16 hours. A lacquer leaf extract was obtained. At this time, the ethanol concentration of the mixed solvent was 70%, 40%, or 35%.

With respect to the obtained sumac leaf extract and a solution sample having an ethanol concentration of 35% (concentrated solution of sumac leaf extract) collected for the evaluation of (B) above, the urushiol content, quercitrin content and total polyphenol amount, and the total polyphenol amount, and , DPPH (2,2-Diphenyl-1-picrylhydrazyl) radical scavenging activity and α-glucosidase inhibitory activity are shown in Table 1. Here, the target values of the content of each component were set to 5% by weight or more for quercitrin and 15% by weight or more for total polyphenols. The higher the value shown in Table 1, the higher the activity of the DPPH radical scavenging activity, and the lower the value shown in Table 1, the higher the activity of α-glucosidase inhibitory activity. ..

As shown in Table 1, the concentrated solution of sumac leaf extract with 70% ethanol (ethanol concentration: 35%) has a urushiol content of less than 0.001% by weight, which is extremely small, and has a quercitrin content and total polyphenols. The amount exceeded the target of 5% by weight and 15% by weight. On the other hand, in the unconcentrated sumac leaf extract, when the ethanol concentration is 70%, the quercitrin content and the total polyphenol amount exceed the target values, but the urushiol content is as high as 5.9% by weight, which is as it is. Was not usable as a pharmaceutical raw material. Further, when the ethanol concentration was lowered to 35% or 40%, the urushiol content was reduced to some extent, but the quercitrin content and the total polyphenol amount were also reduced, and the DPPH radical scavenging activity and the α-glucosidase inhibitory activity were also lowered. In particular, the α-glucosidase inhibitory effect was significantly lower than that of the concentrated solution of sumac leaf extract.
From the above results, the production method of the present invention, which concentrates the extract of sumac leaves in a water / alcohol mixed solvent, can easily and efficiently remove only urushiol, which is an allergic component, and is a useful component. It was found that it has the characteristic that the amount of quercitrin and polyphenol can be maintained without decreasing, and is extremely superior to the method without the concentration step.

[Evaluation of activity]
(C) Evaluation of Antioxidant Activity With respect to the dry powder of sumac leaf extract (composition 1) obtained in Example 1, the oxygen radical absorption capacity (ORAC) was measured to evaluate the antioxidant activity, and the total amount of polyphenols was catechin. Measured in equivalents. The results are shown in Table 2.

(D) Evaluation of Weight Gain Suppressing Action, Intraperitoneal Fat Accumulation Suppressing Action, and Blood Leptin Amount Increase Suppressing Action Regarding the dried urushi leaf extract powder (composition 1) obtained in Example 1, the weight gain suppressing action and the intraperitoneal fat The accumulation inhibitory effect and the blood leptin amount increase inhibitory effect were evaluated. The test was performed on 6 week old male C57BL / 6J mice. Specifically, 18 C57BL / 6J mice (purchased from Nippon SLC Co., Ltd.) were divided into three groups I, II, and III, and group I was a high-fat feed (manufactured by Oriental Yeast Co., Ltd .: HFD-60). , Group II was given a high-fat feed containing 1% by weight of composition 1, and Group III was given a high-fat feed containing 2% by weight of composition 1, and the body weight was measured twice a week. On the 56th day from the start of the test, the amount of intraperitoneal fat was measured and blood was collected to measure the amount of leptin in the blood. Statistical processing was performed by the Dunnett method, and the significance level was set to 0.05. The measurement result of the body weight is shown in FIG. 2, the measurement result of the abdominal fat mass is shown in FIG. 3, and the measurement result of the blood leptin amount is shown in FIG. In FIGS. 2-4, error bars represent standard errors. The amount of leptin in blood was measured by the ELISA method.
As shown in FIGS. 2 to 4, a tendency of suppressing weight gain was observed in Group II and a significant effect of suppressing weight gain was observed in Group III as compared with Group I. In addition, Group III showed a tendency to suppress the accumulation of intraperitoneal fat as compared with Group I, and a significant effect of suppressing the increase in blood leptin level was observed.

According to the production method of the present invention, the urushiol content of the sumac extract can be significantly reduced by a simple step, so that the composition contains the active ingredient of sumac and the content of urushiol that induces allergies is reduced. Goods can be manufactured efficiently at low cost. Therefore, the present invention has high industrial applicability.

Claims (16)

1. A method for producing a urushiol content-reducing composition, which comprises a step of concentrating a solution of raw sumac in a mixed solvent of water and alcohol to obtain a concentrated solution in which the volume of the solution is halved or less.
   Urushiol content, which comprises a step of concentrating a solution containing a mixed solvent of water and alcohol and a component of sumac extracted in the mixed solvent to obtain a concentrated solution in which the volume of the solution is halved or less. Method for producing reduced composition.
2. The manufacturing method according to claim 1, wherein the sumac is a leaf of sumac.
3. The production method according to claim 1 or 2, wherein when the alcohol concentration before concentration is a% by volume and the alcohol concentration after concentration is b% by volume, ab is 10% by volume or more.
4. The production method according to any one of claims 1 to 3, wherein the concentration reduces the alcohol concentration of the solution to less than 50% by volume.
5. The production method according to any one of claims 1 to 4, wherein the concentration is carried out under reduced pressure.
6. The production method according to any one of claims 1 to 5, wherein the concentration is carried out at 20 to 50 ° C.
7. The production method according to any one of claims 1 to 6, wherein urushiol contained in the solution is precipitated by the concentration.
8. The production method according to any one of claims 1 to 7, wherein the concentration reduces the concentration of urushiol contained in the solution to less than 1/10.
9. The production method according to any one of claims 1 to 8, wherein the urushiol concentration of the concentrated solution is less than 0.1% by weight.
10. The production method according to any one of claims 1 to 9, wherein 80% by weight or more of kercitrin contained in the solution before concentration remains in the concentrated solution after concentration.
11. The production method according to any one of claims 1 to 10, further comprising a step of drying the concentrated solution into a powder.
12. A composition produced by the production method according to any one of claims 1 to 11.
13. The composition according to claim 12, further containing one or more selected from the group consisting of elm, oyster, lychee, litchi and keninjin.
14. An anti-obesity agent comprising the composition according to claim 12 or 13.
15. A fat accumulation inhibitor containing the composition according to claim 12 or 13.
16. An agent for suppressing an increase in the amount of leptin in blood, which comprises the composition according to claim 12 or 13.

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