JP2001335505A - Antistress agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antistress agent safe and effective in humans or animals. SOLUTION: This antistress agent comprises an extract derived from sugarcanes. A food and a feed comprise the antistress agent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an anti-stress agent for humans or animals. The invention also relates to a food or feed comprising an anti-stress agent for humans or animals.

[0002]

2. Description of the Related Art The cause of stress is mental,
There are various things such as overwork, infectious disease, pregnancy, drug administration, and as a result, it is known that a wide variety of diseases occur.

For example, in humans and companion animals, stress affects a wide range of the immune system, nervous system, digestive system, circulatory system and the like. Opportunistic infections and carcinogenesis due to decreased immunity in the immune system, neuropathy such as depression and epilepsy in the nervous system, gastric and duodenal ulcers and irritable bowel syndrome in the digestive system, hypertension and ischemic heart disease in the circulatory system Etc. are reported to be caused by stress.

[0004] Also, in livestock and farmed fish, overcrowding, insufficient management of temperature and humidity, excessive feeding and transport of concentrated feed in the livestock barn and cages cause stress, poor growth, decreased spawning rate, and meat quality. It causes diseases such as lowering, transportation fever, pneumonia, diarrhea, and gastric ulcer.

[0005] It is generally known that the thymus and spleen atrophy due to stress, and the theory that this is a physiological response is now mainstream (Abo, T.,
Therapy, 81, 1026-1032, 1999). In other words, the thymus is mainly responsible for the immune response to foreign antigens, but during stress, the system that handles internal abnormalities becomes relatively important. Therefore, thymus atrophy is often used as an index for observing a stress state experimentally.

[0006] Conventionally, as an anti-stress substance, an anti-stress composition containing β-carotene as an active ingredient (Japanese Patent Application Laid-Open No.
No. 65068), an anti-stress agent containing L-athenin as an active ingredient (Japanese Patent Application Laid-Open No. 6-100442), a stress-responsive food prepared by mixing a plurality of aspirating crude drugs with spirulina (Japanese Patent Application Laid-Open No. 9-202), astaxanthin as well as/
Or an anti-stress composition comprising an ester thereof as an active ingredient (JP-A-9-124470), a high glutathione content yeast or a mixture of glutathione and yeast, for relieving stress in livestock and poultry. Feed (JP-A-9-262061), an anti-stress agent containing a tripeptide having angiotensin-converting enzyme inhibitory activity and / or a salt thereof as an active ingredient (JP-A-11-206201)
No. 100328).

[0007]

However, some of the conventionally known anti-stress agents do not always exhibit sufficient effects or are not stable. Further, to cope with various symptoms due to a wide variety of causes, there is a long-awaited need to develop new anti-stress agents. Therefore, an object of the present invention is to provide a safe and highly effective novel anti-stress agent.

[0008]

Means for Solving the Problems The present inventors have conducted intensive studies in view of these backgrounds, and as a result, have found that a sugarcane-derived extract has a safe and effective anti-stress effect, and completed the present invention. Was. That is, the present invention is an anti-stress agent comprising an extract derived from cane sugar as an active ingredient.

The present invention is particularly directed to an anti-stress composition comprising a fraction obtained by subjecting a raw material selected from sugar cane juice, a solvent extract of cane sugar and molasses derived from cane sugar to column chromatography using a fixed carrier as an active ingredient. Agent.

[0010] Alternatively, the present invention is an anti-stress agent comprising, as an active ingredient, an extract obtained by extracting sugarcane-derived bagasse as a raw material with water, a hydrophilic solvent, or a mixture thereof.

Although sugar cane is originally cultivated as a raw material plant to obtain sucrose, the present invention extracts an extract from a raw material selected from cane juice, molasses derived from cane sugar and a solvent extract of cane sugar by column chromatography. Since the method or the method of extracting sugarcane, which is normally discarded, from bagasse, which is a residue obtained by pressing the sugarcane, is employed, even if the extract of the present invention is extracted, sucrose can be recovered from the sugarcane juice at the same yield as before. It does not interfere with conventional sucrose production. That is, since the extract of the present invention can be obtained from the portion of sugarcane except for sucrose, it can be produced at low cost and also makes effective use of resources.

In the present invention, "anti-stress agent"
Relieves stress caused by various factors,
It also refers to an agent for preventing, treating, or ameliorating various symptoms and diseases induced in the immune system, nervous system, digestive system, circulatory system, and the like by stress.

In the present invention, an animal means a vertebrate other than a human, and includes mammals, birds and fish. For example, livestock such as cows, pigs, horses, goats, sheep, poultry such as chickens, quails, ducks, etc. And companion animals such as cats.

In the present invention, the sugarcane-derived extract is
It is an extract obtained from sugar cane as a raw material.

[0015] In one embodiment, the extract derived from sugar cane is a raw material selected from sugar cane juice, a solvent extract of sugar cane and molasses derived from sugar cane (hereinafter, simply referred to as a raw material) using a fixed carrier. It is a fraction obtained by column chromatography. More preferably, the raw material is
A fraction obtained by passing the solution through a column filled with a synthetic adsorbent as an immobilized carrier and eluting the component adsorbed by the synthetic adsorbent with a solvent selected from water, methanol, ethanol and a mixture thereof. Alternatively, of the fractions obtained by separating the raw materials by column chromatography using a difference in affinity in a column packed with an ion exchange resin as a fixed carrier, a wavelength of 420 n
m is a fraction that absorbs light.

Heretofore, the evaluation of the color value of raw materials, intermediate products and products derived from sugar cane has been carried out based on the absorbance of light having a wavelength of 420 nm. Since the absorbance is slightly affected by the pH of the sample, the absorbance is usually measured after adjusting the pH to around neutral. In the present invention, the absorbance is measured after adjusting the pH of the sample to a range of 6 to 8. As described in the Examples, 0.25 g of the lyophilized powder of the obtained fraction was mixed with 0.5 mM phosphate buffer (p
H7.5) to make the total amount 100 ml, and when measuring the absorbance at a wavelength of 420 nm using a 1 cm cell,
The high effect of the present invention is recognized in the fraction having an absorbance of 0.8 or more. However, it is not known whether the absorbance at a wavelength of 420 nm is a value indicating the amount of the pigment derived from sugar cane and is a value indicating the amount of the active ingredient itself. Due to the difference in the absolute amount of the pigment contained, the effect and the absorbance are not always proportional to various extracts derived from different sugarcane.
Measure the absorbance of multiple fractions obtained from one raw material,
The fraction having a relatively high absorbance is the fraction of the present invention.

When a column is filled with a synthetic adsorbent as an immobilized carrier and column chromatography is performed, the affinity of the active ingredient having an anti-stress effect to the synthetic adsorbent is very strong, so that when the raw material is passed through the column. The active ingredient is adsorbed on the synthetic adsorbent. Thereafter, when elution is performed with a solvent, the components adsorbed on the synthetic adsorbent are desorbed and eluted. On the other hand, when an ion exchange resin is used as the fixed carrier,
Since there is a difference in the affinity between the active ingredient and the other components for the ion exchange resin, the raw material is supplied to the column, and then water is flowed as an eluent to reduce the elution rate between the active ingredient and the other components. The two can be separated based on the differences. The affinity between the active ingredient for the anti-stress effect and the resin is not so strong as that of the adsorption.

[0018] Alternatively, in another embodiment, the sugarcane-derived extract is obtained by converting sugarcane-derived bagasse into water, a hydrophilic solvent,
And an extract obtained by elution with a solution selected from a mixture thereof, and more preferably, bagasse, which is a residue obtained by pressing sugar juice from cane sugar, with water, ethanol,
And an extract obtained by extraction with a solution selected from a mixture thereof.

The sugar cane juice according to the present invention is a pressed juice obtained by pressing sugar cane (sugar cane), a leached juice obtained by leaching sugar cane with water, or a lime-treated clean juice obtained at a raw sugar manufacturing plant. Includes concentrated juice.

The solvent extract of sugar cane in the present invention means an extract obtained by concentrating and drying a solution obtained by extracting sugar cane with a general-purpose organic solvent and then re-dissolving it in water. Examples of the organic solvent immediately above include alcohols such as methanol and ethanol, and these may be used alone or in combination. Further, these solvents and water may be used in combination.

The molasses derived from cane sugar in the present invention means a molasses obtained by subjecting a mixture of a sugar crystal obtained in a crystallization step and a mother liquor to centrifugation and separating the mixture from the sugar crystals. The first honey in the factory, the second honey, sugar waste honey,
And molasses at a refined sugar production plant, 1st to 7th molasses, refined waste honey and the like. In addition, those obtained by subjecting molasses to a desugaring treatment, such as a separated liquid obtained by performing alcohol fermentation using these molasses as a raw material, can also be used.

In the present invention, bagasse typically means bagasse discharged in a sugar-making process in a sugar factory. In addition, the bagasse discharged in the sugar-making process in the raw sugar factory mentioned here includes not only the final bagasse that has exited the final press but also the shredded sugar cane that has been cut into the subsequent press including the first press. . Preferably, bagasse discharged after squeezing the sugar juice by a squeezing step in a raw sugar factory is used. The bagasse discharged from the squeezing process has different moisture, sugar content and composition ratio depending on the type of sugar cane, harvest time, and the like. In the present invention, these bagasses can be used arbitrarily. Further, similarly to the raw sugar factory, for example, bagasse remaining after the cane squeezing discharged in the brown sugar factory may be used. Alternatively, in a small-scale implementation at a laboratory level, bagasse obtained by squeezing a sugar solution from sugar cane may be used.

Such a sugarcane-derived extract can be more specifically obtained, for example, as follows.

First, a method for obtaining by column chromatography will be described.

Sugar cane juice, a solvent extract of cane sugar, or molasses derived from cane sugar (hereinafter sometimes referred to simply as a raw material) is passed through a column filled with a fixed carrier. The above raw materials can be used as they are or adjusted to an arbitrary concentration with water. In addition, in order to remove foreign substances, it is preferable to filter the raw material before the treatment with the column. The method of filtration is not particularly limited, and means such as screen filtration, diatomaceous earth filtration, microfiltration, and ultrafiltration widely used in the food industry can be preferably used.

As the fixed carrier, a synthetic adsorbent and an ion exchange resin are preferable.

First, a preferred embodiment of the method using a synthetic adsorbent as a fixed carrier is as follows.

As the synthetic adsorbent, preferably, an organic resin can be used, and for example, an aromatic resin, an acrylic methacrylic resin, an acrylonitrile aliphatic resin and the like can be used. More preferably an aromatic resin,
In particular, an unsubstituted aromatic resin can be used. As the synthetic adsorbent, for example, an aromatic resin such as a styrene-divinylbenzene resin can be used, and as the aromatic resin,
For example, a porous resin such as an aromatic resin having a hydrophobic substituent, an unsubstituted aromatic resin, and an aromatic resin obtained by applying a special treatment to an unsubstituted group can be used. More preferably, an aromatic resin that has been subjected to a special treatment of an unsubstituted group can be used. Such synthetic adsorbents are commercially available, for example, Diaion ™ HP-10, HP-20, HP-21, H
P-30, HP-40, HP-50 (above, unsubstituted aromatic resin, manufactured by Mitsubishi Chemical Corporation); SP-82
5, SP-800, SP-850, SP-875, SP
-70, SP-700 (above, an aromatic resin obtained by subjecting an unsubstituted group to a special treatment, manufactured by Mitsubishi Chemical Corporation); SP-9
00 (aromatic resin, manufactured by Mitsubishi Chemical Corporation); Amberlite (trademark) XAD-2, XAD-4, XAD-1
6, XAD-2000 (above, aromatic resin, manufactured by Organo Co., Ltd.); Diaion (trademark) SP-205, S
P-206, SP-207 (above, aromatic resin having a hydrophobic substituent, manufactured by Mitsubishi Chemical Corporation); HP-2M
G, EX-0021 (above, aromatic resin having a hydrophobic substituent, manufactured by Mitsubishi Chemical Corporation); Amberlite (trademark), XAD-7, XAD-8 (above, acrylic ester resin, stock) Diaion (trademark) HP1MG, HP2MG (above, acrylic acid methacrylic resin, manufactured by Mitsubishi Chemical Corporation); Sephadex (trademark) LH20, LH60 (above, derivatives of cross-linked dextran, Amersham Pharmacia Biotech Co., Ltd.) Manufactured). Among them, SP-850 is particularly preferred.

The amount of the fixed carrier varies depending on the size of the column, the type of the solvent, the type of the fixed carrier, and the like. The wet volume is preferably 0.01 to 5 times the solid content of the raw material.

By passing the raw material through the column, the component having an anti-stress effect in the raw material is adsorbed on the fixed carrier, and most of sucrose, glucose, fructose and inorganic salts flow out as it is.

The components adsorbed on the fixed carrier are eluted by the solvent. Here, in order to elute the component having the anti-stress effect efficiently, it is preferable to sufficiently wash out the remaining sucrose, glucose, fructose and inorganic salts by water washing. Thereby, the adsorbed component having the target anti-stress effect can be efficiently collected. The elution solvent is preferably selected from water, methanol, ethanol and a mixture thereof, more preferably a mixed solvent of water and alcohol, particularly preferably a mixed solvent of ethanol and water. In order to efficiently elute components having the desired effect at room temperature,
A 50 / 50-60 / 40 (vol / vol) ethanol-water mixed solvent is preferred. By further increasing the column temperature, the ethanol mixture ratio of the ethanol-water mixture solvent can be reduced, and components having an anti-stress effect can be eluted. In this case, the inside of the column is in a normal pressure or a pressurized state. Components having an anti-stress effect are
Present in the fraction eluted with the solvent. The elution rate is not particularly limited since it varies depending on the size of the column, the type of the solvent, the type of the fixed carrier, and the like.
hr ^-1 is preferred. In addition, SV (Space velocity, space velocity) is a unit of how many times the amount of liquid per unit volume of resin flows per hour.

The component having the anti-stress effect can be preferably obtained as follows, but is not limited to the following. That is, 0.01 to the solid content of the raw material
After passing the raw material at a column temperature of 60 to 97 ° C. through a column packed with a 5-fold wet volume of an unsubstituted aromatic resin, the column is washed with water, and the components adsorbed on the column are then absorbed. At 50 / 50-60 at a column temperature of 20-40 ° C.
/ 40 (vol / vol) ethanol-water mixed solvent, and the amount of eluate collected from the start of elution is 4% of the resin.
Collect fractions eluting within double wet volume.

On the other hand, a preferred embodiment of the method using an ion exchange resin as the fixed carrier is as follows.

The ion exchange resin is classified into a cation exchange resin and an anion exchange resin from the viewpoint of ion exchange properties. In the present invention, a cation exchange resin can be preferably used. More preferably, a strongly acidic type sodium ion type or potassium ion type cation exchange resin can be used. In addition, from the viewpoint of the form of the resin, the ion exchange resin is a gel type resin, a porous type, a microporous type,
Although it is classified into a porous resin such as a high-porous resin, a gel-type ion exchange resin can be preferably used in the present invention. More preferably, a strongly acidic gel-type cation exchange resin of sodium ion type or potassium ion type can be used. Such ion exchange resins are commercially available, for example, SK1B,
SK104, SK110, SK112, SK116 (Mitsubishi Chemical Corporation), UBK530, UBK550
(Above, for chromatographic separation, Mitsubishi Chemical Corporation), Amberlite (trademark) as Amberlite IR120
BN, IR124, XT1006, IR118, Amber List 31, Amberlite CG1 for chromatograph
20, CG6000 (above, Organo Corporation), Dowex (trademark) type, HCR-S, HCR-W
2, HGR-W2, Monosphere 650C, Marathon C600, 50W × 2, 50W × 4, 50W × 8 (Dow Chemical Japan Co., Ltd.), Muromak 50W
X (Muromachi Chemical Co., Ltd.), as Purolite (trademark) type, C-100E, C-100, C-100 × 1
0, C-120E, PCR433, PCR563K, P
CR822, PCR833, PCR866, PCR88
3, PCR 892, PCR 945 (all of which are described above by MP Ionics). Above all, U
The BK series is particularly preferred.

The amount of the fixed carrier varies depending on the size of the column, the type of the fixed carrier, and the like. It is preferably 2 to 10,000 times, more preferably 5 to 10,000 times the solid content of the raw material.
~ 500 times the wet volume.

The raw material is passed through the above column, and then subjected to chromatography using water as an eluent, and a fraction that absorbs light having a wavelength of 420 nm is collected from a large number of obtained fractions to obtain an objective extract. Can be obtained. In the following:
This method is sometimes called ion chromatography separation.

The conditions for passing the liquid vary depending on the composition of the raw materials and the type of the fixed carrier. In the case of a single-column batch separation method using degassed water as an eluent, the flow rate is SV = 0.
3 to 1.0 hr ^-1 , sample supply is 1 to 20
%, And the temperature is preferably 40 to 70 ° C. For each of the fractions obtained by this separation method, the absorption at a wavelength of 420 nm, the electric conductivity (a measure of the amount of salt), and the concentrations of sucrose, glucose and fructose are analyzed. Peaks appear at 420 nm in absorbance peak, electrical conductivity peak, sucrose and reducing sugar peaks in this order. Fractions corresponding to fractions 3 to 14 in FIG. 1 described below are collected as fractions absorbing light having a wavelength of 420 nm. Particularly, fractions 3 to 8 are preferable. Further, for fractions 17 to 30 after sucrose was completely discharged, although the absorbance at a wavelength of 420 measured using the fractions as they were was low, a sample obtained by concentrating them together was sample 8 of Production Example 8. As shown, the absorbance is high and an anti-stress effect is also observed. In other words, fractions 17 to 30 have low concentrations and are not suitable for efficiently collecting as solids, but the purity of the active ingredient per solid is relatively high. Therefore, the whole non-sucrose fraction (combination of fractions 1 to 8 and / or fractions 17 to 30) has a concentration of the active ingredient lower than that of only fractions 3 to 8, but similarly as an extract of the present invention. Can be used. In the case of the simulated moving bed continuous separation method,
Since the supply amount of the raw material liquid, the flow rate of the eluent, and the flow rate of extracting each fraction are set according to the composition of the raw material, the type of the fixed carrier, and the amount of the resin, general flow conditions cannot be indicated.

The composition of this fraction obtained by a simulated moving bed continuous separation method using a second honey as a raw material in a raw sugar factory varies depending on the type of raw material and the separation ability of the ion exchange resin. Is 6% or less, the non-sugar content is 90% or more, and the apparent pure sugar ratio is 10%. The apparent pure sugar ratio is a percentage of the sugar content (direct optical rotation with respect to the specified amount of pure sucrose measured by a sugar tester) to the Brix (Bx.) Content (percentage of solid content measured by a Brix meter).

The fraction obtained by the single-column batch separation method has a polyphenol content of about 5 per solid (freeze-dried solid) in the case of a fraction obtained using the second honey as a raw material.
%, Electrical conductivity ash (salt) is about 44.7%, and sugar is about 5%.

The substance which is the active ingredient of the anti-stress effect is
Although it is apparent that the active ingredient itself has an absorption at 420 nm, it is apparent that the active ingredient itself has an absorption at 420 nm.

Next, a method for obtaining an extract derived from sugar cane by extracting bagasse will be described.

The bagasse extract is obtained by extracting bagasse with a solution selected from the group consisting of water, a hydrophilic solvent, and a mixture thereof. As the hydrophilic solvent, for example, lower alcohols such as methanol and ethanol, ketones such as acetone, and acetates such as methyl acetate and ethyl acetate can be used. More preferably, ethanol is used as the hydrophilic solvent. Preferred solvents for the extraction are in a ratio up to 60/40 by volume,
More preferably, it is an ethanol-water mixed solvent containing ethanol at a ratio of 50/50 or less by volume. The extraction temperature is
For efficient extraction, 50 to 100 ° C is preferable. The extraction time varies depending on the raw material, type, state and the like of bagasse, but is usually 1 to 3 hours. As the extraction method, a general versatile method can be used, and examples thereof include a method of extracting the bagasse and the extraction solvent together in a container, a method of extracting by circulating the extraction solvent, and a method of extracting continuously. . As a device for the extraction, for example, a Desmet-type extruder, a Lulugi-type extruder, or the like can be arbitrarily used. Since the extract extracted from bagasse has a high sugar content, the sugar may be removed by subjecting the bagasse extract to the same column chromatography treatment as described above.

The extract obtained from sugar cane by various methods as described above can be concentrated by conventional means (removal of solvent under reduced pressure, freeze-drying, etc.) to obtain a component having the effect of the present invention. it can. The thus obtained component having an anti-stress effect can be stored in a liquid or powder form concentrated to a solid content of 20% or more. In the case of preservation, especially in the case of a liquid state, it is preferable to preserve it in a frozen state or in an alcoholic state in order to prevent putrefaction.

The anti-stress agent according to the present invention is effective for prevention, treatment and improvement of various stresses regardless of mental stress and physical stress. It is also effective in preventing, treating, and improving various diseases caused by stress.

The timing and frequency of administration of the anti-stress agent according to the present invention are appropriately selected depending on the type of stress, the type of symptoms or disease caused by stress, and are not particularly limited.

The dose of the anti-stress agent of the present invention can be appropriately determined according to the purpose, and is not particularly limited. When administered orally, it is usually 1 to 1,000 mg / kg body weight, preferably 50 to 1,000 mg / kg body weight per day.

The administration form of the extract derived from sugar cane according to the present invention is not particularly limited, and examples thereof include oral, intravenous, intramuscular, subcutaneous, intradermal, intraperitoneal, rectal, sublingual, transdermal, ophthalmic, etc. Can be administered by any method.

The anti-stress agent according to the present invention may be administered as it is, or after adsorbed on a commonly used carrier for pharmaceutical preparations, it can be converted into a solid preparation or a liquid preparation by a known method, or With or without formulation
It can also be mixed with food, feed, drinking water and the like.

When preparing a solid preparation for oral use, the extract contains excipients, binders, binders, disintegrants, lubricants, coloring agents, flavoring agents, antioxidants, solubilizing agents and the like. After the addition, tablets, coated tablets, granules, powders, capsules and the like are made in a usual manner.

Examples of the above-mentioned excipients include starch, corn starch, dextrin, flour, wheat middling, bran, rice bran, rice bran oil cake, soybean meal, soybean powder,
Soybean oil residue, kinako, glucose, lactose, sucrose, maltose, vegetable oil, animal oil, hydrogenated oil, higher saturated fatty acids, other fatty acids, yeast, mannitol, crystalline cellulose, silicon dioxide, silicon dioxide, anhydrous silicon, calcium silicate, silicic acid, monophosphate Calcium hydrogen, tricalcium phosphate, calcium dihydrogen phosphate and the like are used.

As the binder, for example, polyvinylpyrrolidone, ethylcellulose, methylcellulose, gum arabic, tragacanth, hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium alginate, sodium caseinate, sodium carboxymethylcellulose, propylene glycol, sodium polyacrylate and the like are used. .

As the lubricant, for example, magnesium stearate, talc, stearic acid and the like are used.

As coloring agents and flavors, pharmaceuticals, foods,
What is necessary is just to permit addition to a feed, and it does not specifically limit.

As the antioxidant, for example, ascorbic acid, α-tocopherol, ethoxyquin, dibutylhydroxytoluene, butylhydroxyanisole and the like which are permitted to be added to pharmaceuticals, foods and feeds may be used. Tablets and granules can be coated as needed.

When manufacturing an injection preparation, a pH adjusting agent, a buffer, a suspending agent, a solubilizing agent,
Add stabilizer, tonicity agent, antioxidant, preservative, etc.,
It can be manufactured by an ordinary method. At this time, if necessary,
It is also possible to use a lyophilized agent. This injection can be administered intravenously, subcutaneously, intramuscularly, or the like.

Examples of the suspending agent include methylcellulose, polysorbate 80, hydroxyethylcellulose, acacia, tragacanth powder, sodium carboxymethylcellulose, polyoxyethylene sorbitan monolaurate and the like.

Examples of the solubilizer include polyoxyethylene hydrogenated castor oil, polysorbate 80, nicotinamide, and polyoxyethylene sorbitan monolaurate.

As the preservative, for example, methyl parahydroxybenzoate, ethyl paraoxybenzoate, sorbic acid and the like are used.

The present invention also provides a food and feed containing the above-mentioned anti-stress agent. The food and feed may be solid or liquid, and are not particularly limited. Examples of the food include confectionery, soft drinks, functional seasonings, health foods, and the like. Examples of the feed include feed for companion animals such as dog food and cat food, feed for livestock, feed for cultured fish and shellfish, and the like.

[0060]

EXAMPLES The present invention will be described in detail below with reference to specific production examples and examples, but the present invention is not limited to these production examples and examples.

Production Example 1 Approximately 1,000 liters of squeezed juice (18.6% solids) of sugar cane obtained in the production process of a raw sugar production plant was heated to 80 ° C. with a juice heater, and the tube type External filtration (MH25 type,
Filtration treatment was performed with an effective membrane area of 2 m ² × 3, molecular weight cut off of 100,000, manufactured by Daicel Chemical Industries, Ltd. to obtain a treatment liquid of about 750 liters. 15 liters of the synthetic adsorbent SP-850 (trademark, Mitsubishi Chemical Corporation) was packed in a column (column size: inner diameter 17.0 cm, height 100 cm) equipped with a water jacket, and the above-mentioned squeezed juice filtration solution was added thereto. The solution was passed at a flow rate of 75 liters / hour (SV = 5 hr ^-1 ). During the passage of the squeezed juice filtration liquid, water at 65 ° C. was constantly circulated through the water jacket. Next, 45 liters of ion-exchanged water was supplied at a flow rate of 30 liters /
The column was washed for a time (SV = 2 hr ^-1 ).
After washing with ion-exchanged water, the fraction eluted from the column was subjected to detection of saccharides. As a result, a Beflet Brick (Bx.) Meter (Nagoya type, N-1E type) was used. Was confirmed to be about 0. Thereafter, 30 liters of a 55% aqueous ethanol solution (ethanol / water = 55/45 (volume / volume)) as an eluting solvent was passed through the column at a flow rate of 30 liters / hour (SV = 2 hr ^-1 ) to synthesize and adsorb. The component adsorbed on the agent was eluted.
During the elution solvent and the subsequent passage of ion-exchanged water,
Water at 25 ° C. was constantly circulated through the water jacket. The eluate eluted from the column with a 55% aqueous ethanol solution and the eluate obtained by flowing ion-exchanged water for 20 minutes are mixed, concentrated under reduced pressure by a concentrator to about 20 times, and freeze-dried overnight. Thus, 435 g of a brown powder (an extract derived from cane sugar) was obtained.

Production Example 2 Purified juice (solid content 1) obtained in the production process of a raw sugar production plant
(1.5%) A column treatment was carried out in the same manner as in Production Example 1, except that about 620 liters were used without ultrafiltration treatment. The obtained eluate was concentrated under reduced pressure about 20-fold with a concentrator, and then freeze-dried overnight to obtain 210 g of a brownish powder (an extract derived from sugar cane).

Production Example 3 1 kg of dry bagasse obtained at a raw sugar production plant was put in a bag made of nylon net, put into a tank together with the bag, 25 liters of water at 80 ° C. was added, and the mixture was stirred and extracted for 1 hour. The obtained extract was filtered with a cotton filter to remove foreign substances.
The filtrate was concentrated under reduced pressure using a centrifugal thin film concentrator, and then lyophilized overnight to obtain 26.31 g of a brownish powder (an extract derived from sugar cane).

Production Example 4 An extract was extracted in the same manner as in Production Example 3 except that 2 kg of raw bagasse obtained at a raw sugar production plant was used. The obtained extract was filtered through a cotton filter to remove foreign substances, half of which was concentrated under reduced pressure using a centrifugal thin film concentrator, and then freeze-dried overnight to obtain 16.24 g of a brown powder (sugar-derived extract).
I got The filtrate from which the remaining foreign matter was removed was 0.45.
After filtration with a μm cellulose acetate filter for sterilization, the mixture was concentrated under reduced pressure with a centrifugal thin film concentrator, and then lyophilized overnight to obtain 15.45 g of a brown powder (an extract derived from sugar cane).

Production Example 5 350 g of dried bagasse obtained in a sugar factory was put in a nylon net bag, and the bag was put in a stainless steel pan with a bag.
0/50 (vol / vol) ethanol-water mixed solvent 5.2
Five liters were added and extracted for 2 hours at room temperature. The obtained extract was filtered through filter paper (Toyo Filter Paper No. 2, manufactured by Advantech Toyo Co., Ltd.) to remove foreign substances, and the filtrate was concentrated under reduced pressure with an evaporator, and then lyophilized overnight to give 6.72.
g of a brownish powder (an extract derived from cane sugar) was obtained.

Production Example 6 500 g of dry bagasse obtained in a sugar factory was put in a nylon net bag, and the bag was put into a stainless steel pan with a bag.
0/50 (vol / vol) ethanol-water mixed solvent 7.5
One liter was added and extracted at room temperature for 24 hours. The obtained extract was filtered with filter paper (Toyo Filter Paper No. 2, manufactured by Advantech Toyo Co., Ltd.) to remove foreign substances, and the filtrate was concentrated under reduced pressure with an evaporator, and then lyophilized overnight to give 9.95.
g of a brownish powder (an extract derived from cane sugar) was obtained.

Production Example 7 In a raw sugar factory, sucrose crystals were collected twice in a crystal can,
Ion exchange column chromatography was performed by a pseudo moving bed continuous separation method using second honey, which is a honey whose crystals were removed by centrifugation, as a raw material, using a separation sugar filled with a cation exchange resin. Since the steps from the preparation of the raw materials to the ion-exchange chromatography separation are performed continuously, the solids concentration and composition of the liquid in each step fluctuate slightly with time, but the following concentrations and compositions are measured values in steady-state operation. . The second honey had a Brix (Bx.) Of about 85. Since this concentration was high for performing column chromatography, it was diluted to about 50 Brix. Slaked lime and sodium carbonate were added thereto to aggregate impurities, and diatomaceous earth filtration was performed. The resulting filtrate was Brix 47.3, sugar content (Pol.) 23.6,
The purity ratio (purity) was 49.9 and the reducing sugar content was 2.5%. The filtrate was used as a raw material for ion exchange chromatography. Ion exchange chromatography was performed by a simulated moving bed continuous separation method using UBK530 (Mitsubishi Chemical Corporation) as a cation exchange resin. The separation tower filled with resin is divided into eight sections, and the amount of resin per tower is 6.5 m ³ . Supply and extraction were continuously performed by switching the supply of the raw material liquid and the eluent (water) and the extraction positions of the sucrose fraction and the non-sucrose fraction at regular intervals. The default values at the regular time are a supply flow rate of 3 m ³ / hour, an elution water flow rate of 13.5 m ³ / hour, and a non-sucrose fraction extraction flow rate of 1.
2.13 m ³ / hour, withdrawal flow rate of sucrose fraction 4.37 m ³
/ Hour, switching time was 267 seconds. By this chromatography, a sucrose fraction and a non-sucrose fraction were separated. These are fractions 10 to 1 in FIG.
7, and the combination of fractions 1 to 9 and fractions 18 to 30. The sucrose fraction is about 87% sucrose per solid
(By HPLC analysis) Brix is about 35;
This fraction was mixed with the fresh juice, returned to the sugar-making main step, and an operation of recovering sucrose again was performed. The non-sucrose fraction obtained had a sucrose content of about 0.3% (by HPLC analysis) and a Brix of about 8. The non-sucrose fraction was concentrated by a concentration can, and Brix 40.0, sugar content (Pol.)
2.3, pure sugar ratio (Purity) 5.8, reducing sugar content 5.
4%. This non-sucrose fraction was subjected to lyophilization overnight for use in subsequent tests. 0.25 g of the obtained freeze-dried powder was added to a 0.5 mM phosphate buffer (pH 7.0).
In 5), a 100 ml solution was prepared, and the absorbance at a wavelength of 420 nm was measured. The absorbance was 1.11.

Production Example 8 Using the second honey treated liquid obtained at the raw sugar factory as a raw material, ion exchange column chromatography was performed by a single-column batch separation method. The second honey treating liquid used as a raw material is 2
After the honey is diluted, it is subjected to cleaning treatment with sodium carbonate and diatomaceous earth filtration. The analysis values of this raw material liquid were Brix 47.4, sugar content (Pol.) 23.2, pure sugar rate (Purity) 48.9, and reducing sugar content 3.2%. Using this raw material, fractionation separation by ion exchange chromatography was performed by a single column batch separation method using an FPLC system (manufactured by Pharmacia). Gel type strongly acidic cation exchange resin UBK530, sodium ion type (trade name, Mitsubishi Chemical Corporation) 500 in the column
ml. The column has an inner diameter of 26 mm and a height of 1000
mm, with a flow adapter. The flow conditions are
Using degassed distilled water as an eluent, flow rate SV = 0.5
The reaction was performed at hr ^-1 (4.17 ml / min) at a temperature of 60 ° C. About 25 ml of the raw material was supplied to the column. The fractionation conditions were as follows: collection of the eluate was started 30 minutes after the supply of the raw material, and the collection was performed for 3.6 minutes (about 15 ml / tube) per test tube.
This was collected. Wavelength 420 n for 30 fractions obtained
m, absorbance, electrical conductivity, and sugar concentration were measured.
It was shown to. Here, for the absorbance measurement, 0.5 mM
0.1 ml of each fraction was added to 2 ml of phosphate buffer (pH 7.5).
ml was added to make a sample. For electrical conductivity measurement,
Each fraction was diluted to 0.5% with distilled water to prepare a sample. Sugar concentration was measured by HPLC. In order to analyze each peak, the absorption peak portion at a wavelength of 420 nm was combined into four, the sucrose peak portion into three, and the sucrose peak and subsequent portions into one. That is, fractions 3 and 4 are combined into sample 1, fractions 5 and 6 are combined into sample 2, fractions 7 and 8 are combined into sample 3, fractions 9 and 10
To the sample 4, the fractions 11 and 12 to the sample 5, the fractions 13 and 14 to the sample 6, and the fractions 15 and 16 to the sample 7.
Then, fractions 17 to 30 were combined to obtain Sample 8. Fractions 1 and 2 were discarded because they had few components to elute. Each sample was lyophilized overnight to a powder. 0.25 g of the obtained lyophilized powder was dissolved in 0.5 mM phosphate buffer (pH 7.5) to make 100 ml, and the absorbance at a wavelength of 420 nm was measured. The distribution ratio of freeze-dried solids, conductivity ash (salt), sucrose, glucose and fructose contents, and polyphenol content were also measured. The conductivity ash was calculated from the calibration curve of the relationship between the electric conductivity and the known sulfated ash, and the sucrose, glucose and fructose contents were determined by HPLC analysis. (%). The polyphenol content was measured by the Folin-Ciocalteu method in which a calibration curve was drawn using an aqueous catechin solution as a standard solution, reacted with a phenol reagent, and measured for absorbance at a wavelength of 765 nm, and expressed as catechin-converted values. The distribution ratio of the freeze-dried solid is the ratio (%) of the weight of the solid content of each sample to the sum of the weights of the solid content of all the samples. The liquid separation results of each sample are shown in Table 1 below. The absorbance of Sample 8 was relatively high at 0.86, because the tailed components were collected and concentrated. Sample 8 is a combination of 14 fractions, while the other samples are two fractions together. Therefore, although the absorbance at 420 nm is high, it is inefficient to collect only this fraction as the fraction of the present effect. From the sugar content, it can be seen that samples 1 to 3 and 8 correspond to the non-sucrose fraction and samples 4 to 7 correspond to the sucrose fraction.

[0069]

[Table 1]

Production Example 9 The following operation was performed on the solution of the non-sucrose fraction obtained by repeating Production Example 7 in order to further concentrate the active ingredient. The solution (Bx38.8) of the non-sucrose fraction obtained according to Production Example 7 was diluted 2-fold (w / w) using ion-exchanged water. 50 ml of the synthetic adsorbent SP-850 (trademark, Mitsubishi Chemical Corporation) was added to a water jacketed column (column size; inner diameter 3.9c).
m, height 23.2 cm), and 1.5 liters of the above solution of the non-sucrose fraction was added thereto at a flow rate of 0.25 liter /
The solution was passed at a rate of time (SV = 5 hr ^-1 ). During the passage of the non-sucrose fraction solution, water at 65 ° C. was constantly circulated through the water jacket. Next, 0.15 liter of ion-exchanged water was supplied at a flow rate of 0.25 liter / hour (S
(V = 5 hr ^-1 ) to wash the column. After washing with ion-exchanged water, saccharides were detected in the fraction eluted from the column.
In the total (Nagoya Atago Co., Ltd., type N-1E), it was confirmed that Bx was about 0. Thereafter, 0.1 liter of a 55% aqueous ethanol solution (ethanol / water = 55/45 (volume / volume)) was passed through the column at a flow rate of 0.1 liter / hour (SV = 2 hr ^-1 ) as an elution solvent. The components adsorbed on the synthetic adsorbent were eluted. During the passage of the elution solvent and the subsequent ion-exchanged water flow, water at 65 ° C. was constantly circulated through the water jacket. This operation was repeated 5 times, and the eluate eluted from the column with a 55% aqueous ethanol solution and the ion-exchanged water thereafter were used for 60 times.
The eluates obtained by flowing the mixture for minutes were mixed, concentrated under reduced pressure by a concentrator, and then freeze-dried overnight to obtain 24 g of brown powder (sugar-derived extract).

Acute toxicity test of sugarcane-derived extract The extract powder obtained in Production Example 1 was used for a single oral administration toxicity test in rats. Sixteen male and female Sprague-Dawley SPF rats (Crj: CD (SD)) were obtained at the age of 5 weeks and quarantined and acclimated for about 1 week. The breeding conditions were a temperature of 23 ± 3 ° C., a relative humidity of 50 ± 20%, a ventilation frequency of 1 hour to 15 times, lighting of 12 hours a day, and a solid feed (C
FR-1 (trade name), Oriental Yeast Co., Ltd.) and drinking water were allowed to freely ingest and bred. Thereafter, healthy animals were selected and subjected to the test at the age of 6 weeks. The weight range at the time of administration was 157 to 171 g for males and 123 to 133 g for females. Rats fasted overnight (approximately 16 hours) before administration are given a gavage dose of a sugar cane-derived fraction prepared with distilled water to a predetermined concentration in a fixed dose (10 ml / kg body weight) once. did. Animals in the control group were similarly administered only sterile distilled water. The dose is 200 mg / kg and 100
0 mg / kg, and the control group was added to this for a total of 3 mg / kg.
Groups were used. The number of animals in one group was 5 for both sexes. Refeeding after fasting was started 6 hours after administration, and the animals were bred under the above breeding conditions for 14 days thereafter. The results are shown in Table 2 below.

[0072]

[Table 2]

After 14 days from the administration, no death of rats was observed even at the maximum dose of 1000 mg / kg for both males and females. Therefore, the lethal dose is estimated to exceed 1000 mg / kg. No abnormalities were observed in any of the rats during breeding, and the weights of the males and females in each test solution administration group were almost equivalent to the control group. Anatomical examination showed no abnormalities in organs / tissues of the external surface, head, chest and abdomen in any of the rats. From the above results, it is considered that the toxicity of the extract powder obtained in Production Example 1 in rats after a single oral administration toxicity test was extremely weak.

Example 1 Effect of Cane-Derived Extract on Cold Stress (1) Cold-stressed mice (Slc: ICR, male, 5 weeks old at the start of the experiment, body weight of about 22 to 26 g, 4 animals per group) An effect derived from sugar cane was administered to examine the effect. First, Production Example 1,
The cane-derived extract prepared by the methods shown in 2, 4 and 7 was added in an amount of 0.5 mg so as to give a dose of 500 mg / kg per administration.
dissolved or suspended in 1 L of distilled water for injection, and
Oral administration for 4 consecutive days. Each mouse was then stressed for 4 days. That is, 4 hours on the 4th day from the start of the extract administration, 7 hours on the 5th day, 24 hours on the 6th and 7th days
For hours, each mouse was housed in a cold room at 5 ° C. and subjected to cold stress. After 4 days of stress, the mice were weighed and necropsied, and the liver, spleen and thymus were weighed. The control group was orally administered with 0.5 ml of distilled water for injection, and was similarly subjected to cold stress. Further, as a normal control group, a group which was bred at room temperature and orally administered 0.5 ml of distilled water for injection was also set. The results are shown in Tables 3 and 4.

[0075]

[Table 3]

[0076]

[Table 4]

As is clear from the above table, the weight of the spleen and thymus was reduced in the control group as compared to the normal control group, whereas the decrease was slight in the group to which the cane-derived extract was administered. Was. Therefore, the sugarcane-derived extract of the present invention has an anti-stress effect.

Example 2 Effect of Cane-Derived Extract on Cold Stress (2) Cold-stressed mice (Slc: ICR, male, 5 weeks old at the start of the experiment, body weight of about 24-28 g, 4 animals per group) The effect was examined by administering a sugarcane-derived extract. Using the sugarcane-derived extract prepared by the method shown in Production Example 3, the effect of the sugarcane-derived extract on cold stress was examined in the same manner as in the method shown in Example 1. The results are shown in Tables 5 and 6.

[0079]

[Table 5]

[0080]

[Table 6]

As is clear from the above table, the weight of the spleen and the thymus was reduced in the control group as compared with the normal control group, whereas the reduction was slight in the group to which the cane-derived extract was administered. Was. Therefore, the sugarcane-derived extract of the present invention has an anti-stress effect.

[0082]

Industrial Applicability The extract derived from sugar cane according to the present invention is useful as an anti-stress agent in humans and animals. Since sugar cane has been used as food since ancient times, it is excellent in safety. Further, in the present invention, since the waste portion after the extraction of sucrose is used, it can be manufactured at low cost, resources can be effectively used, and the contribution to industrial development is great.

[0083]

[Brief description of the drawings]

FIG. 1 is a graph showing the absorbance, electric conductivity, and sugar concentration of a fraction obtained by separation using an ion exchange resin in Production Example 8.

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Claims (14)

[Claims] 1. An anti-stress agent comprising an extract derived from sugar cane as an active ingredient. 2. The extract derived from sugar cane is a fraction obtained by treating a raw material selected from cane juice, a solvent extract of cane sugar and molasses derived from cane sugar by column chromatography using a fixed carrier. Item 7. The anti-stress agent according to Item 1. 3. A raw material selected from a sugar cane extract, a sugar cane juice, a solvent extract of sugar cane, and a molasses derived from sugar cane, is passed through a column filled with a synthetic adsorbent as a fixed carrier, and The anti-stress agent according to claim 2, wherein the component is a fraction obtained by extracting the component adsorbed with a solvent selected from water, methanol, ethanol, and a mixture thereof. 4. A column utilizing a difference in affinity between a column in which an extract derived from sugar cane is selected from sugar cane juice, a solvent extract of sugar cane and molasses derived from sugar cane, and which is packed with an ion exchange resin as a fixed carrier. The anti-stress agent according to claim 2, which is a fraction that absorbs light having a wavelength of 420 nm among the fractions obtained by separation by chromatography. 5. The anti-stress agent according to claim 4, wherein the ion exchange resin is a cation exchange resin. 6. The anti-stress agent according to claim 5, wherein the cation exchange resin is a strongly acidic cation exchange resin. 7. The anti-stress agent according to claim 6, wherein the strongly acidic cation exchange resin is of a sodium ion type or a potassium ion type. 8. The method according to claim 4, wherein the ion exchange resin is a gel type.
8. The anti-stress agent according to any one of 7 above. 9. The method according to claim 4, wherein the column chromatography is performed by a simulated moving bed continuous separation method.
The anti-stress agent according to item. 10. The anti-stress agent according to claim 1, wherein the sugarcane-derived extract is obtained by extracting bagasse with water, a hydrophilic solvent, or a mixture thereof. 11. The anti-stress agent according to claim 10, wherein the hydrophilic solvent is ethanol. 12. The method according to claim 12, wherein the hydrophilic solvent for extraction is 60/40.
The anti-stress agent according to claim 10, which is an ethanol-water mixed solvent containing ethanol at a volume ratio or less. (13) A food containing the anti-stress agent according to any one of (1) to (12). 14. A feed containing the anti-stress agent according to any one of claims 1 to 12.