JP5713332B2 - Anti-fatigue composition - Google Patents

Description

  The present invention relates to a composition containing cryptoxanthin and having endurance enhancing action and / or anti-fatigue action on humans and animals, which can be used for foods and drinks, pharmaceuticals, quasi drugs, feeds and the like.

  Fatigue is a condition characterized by reduced work and inefficiency in performance following mental or physical activity, and is said to recover by rest or sleep, but it does not recover but prolongs Then, it may become chronic fatigue syndrome. In addition, for athletes and athletes, increasing endurance and recovering from fatigue lead to improved results, and products for enhancing endurance and anti-fatigue increase efficiency when other people work. Food and drinks and medicines are on sale.

  The same applies to animals, and there is a demand for feeds that are effective in enhancing endurance and anti-fatigue in animals such as racehorses and animals such as pets.

  Until now, various foods and drinks and pharmaceuticals have been sold for the purpose of fatigue recovery. Examples thereof include herbal medicines, herbal extracts, royal jelly, vitamins, minerals, caffeine, amino acids, taurine and the like. Patent Document 1 proposes an anti-fatigue composition comprising a combination of coenzyme Q10, carnitine and an organic acid, and Patent Document 2 proposes an anti-fatigue agent containing a mugwort organic solvent extract as an active ingredient.

  Carotenoids are a kind of pigment component of animals, plants, and microorganisms. These foods often contain specific carotenoids, such as β-carotene for carrots, astaxanthin for crustacean shells, and lycopene for tomatoes. .

  Among them, cryptoxanthin is a carotenoid that is specifically contained in Wenzhou oranges and persimmons, and is one of the six major carotenoids detected in human serum. The usefulness of cryptoxanthin is a carotenoid that is clearly shown to be effective in preventing osteoporosis, diabetes, alcoholic liver injury, rheumatism, arteriosclerosis and the like from animal studies and epidemiological studies.

Patent Document 3 and Patent Document 4 propose that among carotenoids, astaxanthin can be used as an effect of improving the duration of muscle function, fatigue recovery, and fatigue preventive agent. However, the fact that cryptoxanthin has such a muscle endurance enhancement and anti-fatigue effect is not known at all.
JP-A-2005-97161 JP 2006-45082 A Special table 2001-514215 gazette JP 2006-16409 A

  An object of this invention is to provide the new composition with high safety | security which can exhibit endurance enhancement and an anti-fatigue effect.

  As a result of intensive studies in order to solve such problems, the present inventors have found that cryptoxanthin exhibits endurance enhancement and an anti-fatigue effect, and completed the present invention.

  That is, the gist of the present invention is a composition having endurance enhancing action and / or anti-fatigue action characterized by containing cryptoxanthin and / or an ester thereof, preferably cryptoxanthin and / or Alternatively, the ester body is derived from one or more selected from the group consisting of Unshu oranges, Enzyme-treated products of Unshu oranges, and Solvent extracts of Unshu oranges. More preferably, in the present invention described above, flavonoids and / or derivatives thereof are further contained. Preferably, 0.0001 to 3% by mass of cryptoxanthin and / or an ester thereof, flavonoids and / or the derivatives thereof. A derivative containing 0.001 to 30% by mass, and preferably, cryptoxanthin and / or an ester thereof and a flavonoid and / or a derivative thereof are derived from Wenzhou mandarin or an enzyme-treated product of Unshu mandarin. It is.

  Another gist of the present invention is a food / beverage product, pharmaceutical or feed comprising the above-described composition of the present invention.

  According to the present invention, a composition having high safety and excellent endurance enhancement and anti-fatigue effects can be obtained, and a food, beverage, medicine, quasi-drug, feed containing the composition of the present invention, and By doing so, it becomes easy to obtain endurance enhancement and an anti-fatigue effect.

  Hereinafter, the present invention will be described in detail.

  The cryptoxanthin and / or ester thereof in the present invention is not particularly limited, and examples thereof include α-cryptoxanthin, β-cryptoxanthin and fatty acid esters thereof. The fatty acid length is not particularly limited, and examples thereof include fatty acid esters such as lauric acid (C12), myristic acid (C14), palmitic acid (C16), and stearic acid (C18). Moreover, as a supply source of these cryptoxanthin and / or its ester body, those which have a large production amount and are derived from Unshu mandarin, an ancient fruit of Japan, are preferable. Orange juice, etc.), enzyme-treated products of Unshu oranges, and those derived from solvent extract from Unshu oranges.

  The enzyme-treated product of Unshu mandarin in the present invention is obtained by adding an enzyme to Unshu mandarin orange or squeezed residue of Unshu mandarin and strengthening cryptoxanthin (increased concentration). The specific manufacturing method is as follows. Moreover, after squeezing the fruit of Wenzhou mandarin oranges with an inline squeezer, chopper helper squeeze machine, brown squeeze machine, etc., it is filtered or filtered with a paddle type or screw type finisher, etc. It is prepared by collecting the juice residue obtained by sieving or centrifuging.

  The enzyme used for producing the enzyme-treated product of Unshu mandarin is not particularly limited as long as it can decompose organic matter contained in Unshu mandarin, particularly a biopolymer constituting a cell wall, etc. Glucoamylase, cellulase, hemicellulase, pectinase, mannanase, xylanase, protease, peptidase, lipase, maceration enzyme (cell wall degrading enzyme) and the like are used. Among these, saccharide hydrolase cellulase, hemicellulase, pectinase, mannanase, xylanase, and malation enzyme are preferable enzymes because of their high efficiency of strengthening cryptoxanthin, which is an active ingredient. As the enzyme agent to be added, these purified enzymes may be used, or microbial cells or cultures exhibiting these activities, or crudely purified products thereof may be used. Commercially available enzymes can also be used. For example, for pectinase, Sumiteam PX (manufactured by Shinnippon Chemical Co., Ltd.), Sumiteam SPC (manufactured by Shinnippon Chemical Co., Ltd.), pectinex SRL (manufactured by Novozymes Japan Ltd.) ), Sumiteam PMAC (manufactured by Shinnippon Kagaku Kogyo Co., Ltd.), etc., and cellulosin GM5 (manufactured by HIBI), cellulosin HC (manufactured by HIBI), etc. can be used as the hemicellulase. Cellulase Y-NC (manufactured by Yakult Pharmaceutical Co., Ltd.), Entilon MCH (Pinto Kasei Kogyo Co., Ltd.), Cellulase R10 (manufactured by Yakult Pharmaceutical Co., Ltd.), Sumiteam CAP (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Cellulase TP-3 (Kyowa Kasei Co., Ltd.) can be used, As the protease, protease M (manufactured by Amano Enzyme Co., Ltd.), orientase 20A (manufactured by HBI Co., Ltd.) and the like can be used.

These enzymes may be used alone, or two or more kinds of enzymes may be mixed and used.
The amount of the enzyme to be added is not particularly limited, and may be added according to the reactivity of the enzyme. For example, when pectinase is used, it is preferably 1 to 100,000 units, more preferably 10 to 10,000 units, based on 100 g of Wenzhou mandarin oranges.

  After the enzyme is added, the enzyme and the extract are uniformly mixed by stirring or the like to allow the enzyme reaction to proceed. As the reaction temperature at this time, it is preferable to carry out the reaction under conditions where the enzyme is not inactivated and is not easily spoiled, and where cryptoxanthin is not lost. Specifically, the temperature is 0 ° C to 90 ° C, preferably 0 ° C to 80 ° C, and more preferably 0 ° C to 70 ° C. Needless to say, the pH of the reaction is preferably carried out under the optimum conditions of the enzyme, and it is preferably pH 2 to 12, preferably pH 2.5 to 8. The reaction time depends on the squeezed residue used and the amount of the enzyme, but it is usually preferable to set the reaction time to 1 to 48 hours. In the reaction, the reaction may be performed while stirring the reaction product, or may be a stationary reaction.

  After completion of the enzyme treatment, the enzyme-treated reaction product may be used as it is, or a product that has undergone some processing may be used. Specifically, a residue obtained by solid-liquid separation of the reaction product, a product obtained by drying the residue obtained by solid-liquid separation, or a product obtained by drying the reaction product as it is without solid-liquid separation may be used. In addition, a water washing method that can easily remove reaction products such as sugars produced by enzyme treatment by solid-liquid separation of the enzyme-treated reaction product, addition of water, and solid-liquid separation again. When used, it is preferable because impurities can be easily removed.

  The solvent extract of Unshu mandarin in the present invention is obtained by extracting a component containing cryptoxanthin from an enzyme-treated product of Unshu mandarin and / or Unshu mandarin using a solvent and / or supercritical carbon dioxide. When extracting with a solvent, alcohols such as methanol, ethanol, propanol and isopropanol, ketones such as acetone and methyl ethyl ketone, esters such as methyl acetate and ethyl acetate, and halogenated hydrocarbons such as acetonitrile and chloroform Extraction may be performed using aliphatic hydrocarbons such as pentane and hexane, aromatic hydrocarbons such as benzene and toluene, ethers, pyridines, and polyethers. Among these, from the viewpoint of safety, it is preferable to use acetone, ethanol, hexane, and supercritical carbon dioxide, and it is particularly preferable to use ethanol and supercritical carbon dioxide.

  Further, impurities may be subsequently removed. Examples of the impurity removal method include water washing, organic solvent washing, silica gel column, resin column, reverse phase column and the like, activated carbon treatment, partitioning with solvents of different polarity, recrystallization method, vacuum distillation method and the like. .

  The content of cryptoxanthin and / or its ester in the composition of the present invention is 0.0001% by mass to 3% by mass, preferably 0.001% by mass to 2% by mass, and more preferably 0.01% by mass. What is necessary is just to contain-1 mass%.

  In the composition of the present invention, it is desirable to contain a flavonoid and / or a derivative thereof in addition to the above-described cryptoxanthin and / or an ester thereof in terms of enhancing the endurance enhancing action and / or the anti-fatigue action. The flavonoid and / or derivative thereof in the present invention is not particularly limited, and hesperidin, naringin, rutin and α-glucosylated hesperidin, α-glucosylated naringin, α-glucosylated rutin which are derivatives thereof, and Examples include hesperidin, naringin, hesperetin, naringenin, and quercetin, which are aglycones of rutin. Of these, hesperidin and its derivatives, and hesperetin are preferred because of their increased endurance or anti-fatigue effects. As these supply sources, commercially available compounds may be used, or natural products containing these compounds or processed products thereof may be used. For example, flavonoids and / or derivatives thereof are also included in those obtained by enzymatic treatment of Unshu mandarin as described above, and when used, cryptoxanthin and / or its ester form are also included. It is preferable because the composition of the present invention is easily produced.

  In the composition of the present invention, the content of the flavonoid and / or its derivative is 0.001% by mass to 50% by mass, preferably 0.01% by mass to 30% by mass, and more preferably 0.8% by mass. What is necessary is just to contain 1 mass%-20 mass%. The ratio of cryptoxanthin and / or ester thereof to flavonoid and / or derivative thereof is preferably 1: 1 to 1: 10,000, more preferably 1: 2 to 1: 1,000 as a mass ratio. : 10 to 1: 500 is most preferable.

  The measurement of cryptoxanthin used high performance liquid chromatography (HPLC). As an HPLC apparatus, LC-10A manufactured by Shimadzu Corporation was used, a Resolve C18 (φ3.9 × 150 mm) column manufactured by Waters Co. was connected, and a sample to which an equal amount of methanol was added was introduced. The mobile phase was analyzed for content at methanol: ethyl acetate = 7: 3, column temperature 30 ° C., flow rate 1.0 ml / min, detection wavelength 450 nm.

  The flavonoid content was measured using high performance liquid chromatography (HPLC). As the HPLC apparatus, LC-10A manufactured by Shimadzu Corporation was used, an Asahi Pak-50 (φ4.6 × 150 mm) column manufactured by Asahi Kasei Kogyo Co., Ltd. was connected, and a sample dissolved in 0.1N NaOH was diluted 10 times with distilled water and introduced. did. The mobile phase was analyzed for water: acetonitrile = 8: 2, column temperature 30 ° C., flow rate 1.5 ml / min, and detection wavelength 285 nm.

  In the composition of the present invention, a substance having an endurance enhancing or anti-fatigue effect other than cryptoxanthin and flavonoids may be contained. In this case, the blending amount is preferably 0.01% by mass to 90% by mass as the amount of substances other than cryptoxanthin and flavonoids. As substances other than cryptoxanthin and flavonoids that have endurance enhancing or anti-fatigue effects, known substances can be used. For example, taurine, glucuronic acid, glucuronolactone, theanine, caffeine, γ- Aminobutyric acid, capsaicin, polyphenols, catechins, xanthine derivatives, amino acids such as methionine, leucine, arginine, herbal medicines such as ginseng, deer cocoon, cow yellow, ground yellow, eggplant, royal jelly, minerals such as calcium, magnesium, zinc, iron, It is preferable that various vitamins are blended alone or in combination.

  The method of orally ingesting the composition of the present invention may be ingested as it is by the composition of the present invention alone, or powder, tablet, granule, capsule, gel, soft capsule, paste, syrup, suspension, emulsification. It may be processed into liquids and drinks and taken. The amount of intake for oral intake may be selected in a timely manner depending on the age, sex, weight, etc. of the subject. Usually, for adults, 0.01 mg to 1 g of cryptoxanthin per day, flavonoids and / or 0.1 mg to 10 g of the derivative, preferably 0.1 mg to 100 mg of cryptoxanthin, and 1 mg to 100 mg of flavonoid and / or its derivative may be taken. The standard of intake when parenterally ingesting can be 0.01 mg to 100 mg of cryptoxanthin.

  Another invention of the present invention is a food / beverage product, a feed and a medicine containing the composition of the present invention described above. As pharmaceuticals, injections, infusions, powders, tablets, granules, capsules, pills, intestinal solvents, suspensions, syrups, oral liquids, lozenges, emulsions, liquids for external use, poultices, nasal drops, It can be taken in the form of ear drops, eye drops, inhalants, ointments, lotions, suppositories, enteral nutrients and the like. These can be used alone or in combination of two or more depending on the symptoms. The content of the endurance enhancing and / or anti-fatigue composition may be determined in a timely manner in the range of 0.1% to 90%, but the formulation is designed so that the above-mentioned standard of daily intake can be taken. It is preferable.

  The food and drink of the present invention includes all foods and / or beverages such as foods for specified health use, health foods, functional foods and quasi drugs in addition to general foods. The food and / or beverage is not particularly limited. For example, in addition to the above-mentioned pharmaceutical form, bread, udon, buckwheat, rice, and other staple foods, cheese, wiener, sausage, ham, seafood Processed foods, ice cream, cookies, cakes, jelly, pudding, candy, chewing gum, yogurt, gummy, chocolate, biscuits and other sweets, soft drinks, seasonings, alcoholic beverages, energy drinks, coffee, tea And beverages such as milk, fruit juices, and soft drinks. The content of the composition of the present invention may be determined in a timely manner in the range of 0.1% to 70%, but it is preferable to determine the blending amount so that the above-mentioned guideline for daily intake can be ingested. .

  As feed, for example, grains such as corn, wheat, barley and rye, bran such as bran and rice bran, potatoes such as corn gluten meal and corn jam meal, skimmed milk powder, whey and fish meal It can be produced by blending animal feeds such as bone meal, yeasts such as beer yeast, calciums such as calcium phosphate and calcium carbonate, vitamins, fats and oils, amino acids, saccharides and the like. As a form of feed, it can be used for pet food, racehorse / livestock feed, feed for cultured fish, and the like. The content of the composition may be appropriately determined in the range of 0.01 to 20%, and 0.001 mg to 10 mg of cryptoxanthin per kg of body weight per day and 0.01 mg to 100 mg of flavonoids and / or derivatives thereof per day. It is preferable to add to feed raw materials so that it can be ingested.

  Examples of this experiment are described below. In addition, this invention does not limit the range by this Example.

Reference example 1
The residue after squeezing the fruit juice from Wenzhou mandarin orange (mandarin orange juice, moisture content of about 90%), pectinase enzyme agent Sumiteam PX for food processing (manufactured by Shin Nippon Chemical Industry Co., Ltd., unit number: pectinase 5,000u / g, Add 10 g of arabanase 90 u / g) and cellulase, hemicellulase enzyme cellulase Y-NC (manufactured by Yakult Pharmaceutical Co., Ltd., unit number: cellulase 30,000 u / g), stir well and let stand at room temperature for 8 hours. Reaction was performed. This reaction solution is centrifuged, the supernatant is removed, water is added and stirred, the supernatant is removed again by centrifugation, and the drum temperature is set to 110 ° C., rotating at 1 rotation / minute. Dried at speed. In 50 g of the powder pulverized by a pulverizer, β-cryptoxanthin (as a free form) was contained by 0.5 mass% and hesperidin was contained by 15 mass%.

Reference example 2
8 kg of the residue after squeezing the fruit juice from Satsuma mandarin (mandarin orange juice, water content of about 90%) was freeze-dried, 5 L of ethanol was added thereto, and the mixture was stirred for 1 hour to extract β-cryptoxanthin. Filtration was performed to collect the ethanol fraction, and ethanol was distilled off using an evaporator to obtain Unshu orange extract. This extract contained 5% by mass of β-cryptoxanthin (as a free form).

Examples 1 and 2 and Comparative Example 1
Four-week-old Wister female rats were divided into 3 groups (Example 2 group, Comparative Example 1 group), and exercise training was performed on each group. As exercise training, treadmill walking exercise (10 m / min for 5 minutes, 25-35 m / min for 20 minutes, 10 m / min for 5 minutes for a total of 30 minutes) was performed every week for 5 days. In Example 1, 50 mg of the powder containing β-cryptoxanthin produced in Reference Example 1 was orally administered daily, and exercise training was continued for 2 months. In Example 2, 10 mg of the extract containing β-cryptoxanthin produced in Reference Example 2 was orally administered daily, and exercise training was continued for 2 months. In Comparative Example 1, β-cryptoxanthin was not administered orally and exercise training was continued for 2 months.

  In the measurement, the time from the end of the walking exercise training to the time when the rat started to move again was measured for about 3 weeks in the latter half of the exercise training program. (It was used as a measure of fatigue.) After the exercise training program, a similar walking exercise was performed, and venous blood was collected immediately after the exercise. Plasma lactic acid levels were measured and compared to rest values. Therefore, a catheter was placed in the right external jugular vein under halothane anesthesia several days before the experiment.

  Table 1 shows the time required to start the exercise of Example 1, Example 2 and Comparative Example 1, and Table 2 shows the lactic acid value. Apparently, in the group that orally ingested the composition containing β-cryptoxanthin, shortening of the time until the start of exercise and a decrease in the lactic acid level are observed, and it can be seen that there is an increase in endurance and an anti-fatigue effect.

Example 3 and Comparative Example 2
Twelve volunteer men (mean age 33.4 years ± 9.97) were grouped into a test meal group (Example 3) and a placebo group (Comparative Example 2). Using a bicycle ergometer, exercise was performed for 30 minutes at an exercise intensity of 80% load of the maximum heart rate, and blood lactate levels immediately before and after exercise were measured. The blood lactic acid level was measured with Lactate Pro LT-1710 (Arkray).

  In Example 3, the powder containing β-cryptoxanthin produced in Reference Example 1 was processed into a soft capsule based on olive oil, and 100 mg of the powder was ingested every day. In Comparative Example 2, soft capsules containing olive oil without powder were ingested every day. After 5 weeks and 9 weeks from the start of intake, exercise with a bicycle ergometer was performed in the same manner as before the start of intake, and a questionnaire about blood lactate level and subjective exercise intensity (Borg index) was performed.

  FIG. 1 shows the amount of increase in blood lactate before and after exercise, and FIG. 2 shows a comparison between a questionnaire survey after exercise before the start of intake of the Borg index and a survey result after intake. . In Example 3, a decrease in lactic acid level was observed with ingestion, whereas in Comparative Example 2, there was almost no change in lactic acid level, and a change in the Borg index was also decreased in Example 3. In Comparative Example 2, it is clear that there is little change or an increasing tendency. From these facts, it is apparent that the group ingesting the composition containing β-cryptoxanthin has endurance enhancement and anti-fatigue action, both subjectively and from the blood lactic acid level.

It is a figure which shows the change of the raise amount of the blood lactic acid level before and behind the exercise | movement in Example 3 and Comparative Example 2. FIG. It is a figure which shows the change of the subjective exercise intensity (Borg index) in Example 3 and Comparative Example 2.

Claims (1)

An enzyme-treated product containing cryptoxanthin and / or a fatty acid ester thereof, and hesperidin obtained by treating the juice residue of Unshu oranges with an enzyme capable of degrading a polymer constituting the cell wall of Unshu oranges, and / or Or
A solvent extract containing cryptoxanthin and / or a fatty acid ester thereof obtained by extracting the squeezed residue of Unshu mandarin with ethanol, and for enhancing endurance and / or anti-resistance Fatigue medicine.