JPWO2008149838A1 - Novel royal jelly fraction, its production method and use - Google Patents

Abstract

The first task is to provide a royal jelly fraction that retains the useful physiological activity of royal jelly and is less likely to cause allergic symptoms, and the second task is to establish its production method. It is a third object to provide uses for various compositions such as foods and drinks, feeds, feeds, pet foods or cosmetics for external use, such as cosmetics, which are blended with paintings. It contains a royal jelly-derived protein in a ratio of less than 0.05 parts by mass with respect to 1 part by mass of 10-hydroxy-2-decenoic acid, has a useful physiological activity of royal jelly, and 10-hydroxy-2- Royal jelly fraction in which allergenicity to the royal jelly fraction is not detected when the serum of a mouse immunized with a royal jelly fraction containing 45 μg of decenoic acid is determined by passive skin anaphylaxis using rats, and a method for producing the same Solve by application.

Description

  The present invention has the biological activity of royal jelly, for example, the useful physiological activity of royal jelly such as collagen production enhancing action, IL-4 production inhibitory action and testicular function activation action, and allergenicity in raw royal jelly. Reduced novel royal jelly fraction, its production method and use, specifically, 10-hydroxy-2-decenoic acid (hereinafter sometimes simply referred to as “hydroxydecenoic acid” in this specification) 1 A royal jelly fraction containing a royal jelly-derived protein in a ratio of less than 0.05 parts by mass with respect to parts by mass, including the collagen production enhancing action, IL-4 production inhibiting action, testis function activating action, etc. possessed by royal jelly Is immunized with the royal jelly fraction in an amount containing 45 μg of hydroxydecenoic acid. A royal jelly fraction in which allergenicity of the royal jelly fraction is not detected when the mouse serum is determined by passive cutaneous anaphylaxis (hereinafter referred to as “PCA”) using a rat, a method for producing the same, and use thereof It is about.

  Royal jelly is a milky white secretion from the exocrine glands of worker bees that accumulates in the kingdom (queen bee's bunch) in the beehive, and is a jelly-like substance that is fed as a food to the larvae to become queen bees. Its chemical composition varies slightly depending on the place of origin and season, but water content is 65 to 75% by mass, protein 15 to 20% by mass, carbohydrates 10 to 15% by mass, lipids 1.7 to 6% by mass, ash content 0 0.7 to 2% by mass (hereinafter, unless otherwise specified, mass% is simply abbreviated as “%”), and includes organic acids such as hydroxydecenoic acid and various vitamins. Also contains various minerals.

  Royal jelly has been widely used as a human health food for a long time, and recently, it has useful physiological activities on the human body, such as antibacterial action, immune enhancement action, antitumor action, anti-inflammatory action, life extension action, etc. Many reports have been made. However, since royal jelly contains allergenic substances, ingestion may cause allergic symptoms although there are individual differences, and in some cases, severe anaphylactic shock. For the purpose of solving such problems of royal jelly, Japanese Patent Application Laid-Open No. 2002-127715 discloses a method for reducing the allergen by subjecting royal jelly to glycolytic enzyme treatment and proteolytic enzyme treatment. International Publication WO 2004/21803 discloses a royal jelly in which the amount of water-soluble protein is reduced to less than 50% of the total protein amount. However, these royal jelly is still water-soluble protein that shows allergenicity, or the royal jelly treated with glycolytic enzyme and proteolytic enzyme has insufficient degradation of allergenic substances or added enzyme itself. May become a new allergenic substance, and there is still a problem that it is not preferable as a royal jelly taken for the purpose of maintaining health. In addition, as described in JP-A-4-31357, etc., when extracted with a solvent containing a hydrophobic solvent such as ethanol in order to increase the amount of hydroxydecenoic acid in the royal jelly extract, even when the protein is removed Depending on the composition, there is a problem that the blending amount is limited.

  The present invention has been made in view of such circumstances, and provides a royal jelly fraction that retains the useful physiological activity of royal jelly and has reduced allergenicity and is less likely to cause allergic symptoms. This is the first issue, the establishment of its production method is the second issue, and various types of skin external preparations such as foods, feeds, feeds, pet foods, cosmetics, etc., containing the fractions. The third object is to provide a use for the composition.

  In order to solve the above-mentioned problems, the present inventors have obtained a high-quality royal jelly fraction that has reduced allergenicity and retains the inherently useful physiological activity of royal jelly, and is less likely to cause allergic symptoms. We have conducted extensive research on manufacturing methods. As a result, when the pH of the royal jelly collected from the honeybee kingdom (hereinafter sometimes simply referred to as “raw royal jelly” in this specification) was used as a raw material, the pH was usually 3. It was found to be in the range of 5 to 4.5. On the other hand, the hydroxydecenoic acid contained in raw royal jelly changes its solubility by changing the pH, and improves the solubility by raising the pH, and at the same time, a purification step for reducing the polymer fraction. It has been found that the yield of hydroxydecenoic acid can be improved by increasing the pH. Specifically, the raw royal jelly is diluted by adding water, and preferably, an alkaline agent is further added to adjust the pH to 5.0 or higher, preferably 5.5 or higher, more preferably 5.5 to 12. 0, more preferably 6.0 to 9.5, particularly preferably 6.0 to 8.5, and then the polymer fraction is removed by a method such as filtration through an ultrafiltration membrane or gel filtration. Preferably, the hydroxydecene obtained by removing a fraction having a molecular weight of 30,000 daltons or more, more preferably a fraction having a molecular weight of 10,000 daltons or more, particularly preferably a fraction having a molecular weight of 6,000 daltons or more. A royal jelly fraction (hereinafter, simply referred to as “royal jelly fraction”) containing a royal jelly-derived protein in a ratio of less than 0.05 parts by mass with respect to 1 part by mass of the acid. The existing purified royal jelly in which the water-soluble protein disclosed in Royal Jelly or International Publication No. WO2004 / 21803 is reduced to 50% or less of the total protein mass (hereinafter simply referred to as “existing purified royal jelly” in the present specification) It has been found that the allergenicity is remarkably reduced as compared with other cases. In addition, the above-described royal jelly fraction has the useful physiological activity inherent to raw royal jelly, and has the effect of being less susceptible to turbidity and precipitation even when blended in beverages, etc. Was completed. That is, the present invention is a royal jelly fraction containing a royal jelly-derived protein in a ratio of less than 0.05 parts by mass with respect to 1 part by mass of hydroxydecenoic acid in the royal jelly fraction. Royal jelly fraction with reduced physiological activity and allergenicity, and foods, feeds, feeds, pet foods, cosmetics, pharmaceuticals, and quasi-drugs containing this fraction Moreover, said subject is solved by providing the use to miscellaneous goods composition etc.

  The royal jelly fraction of the present invention retains the original useful physiological functions of the royal jelly, and has reduced allergenicity compared to raw royal jelly and existing purified royal jelly, so the royal jelly fraction, And the composition of foods and drinks, feeds, feeds, pet foods, cosmetics, pharmaceuticals, quasi-drugs, miscellaneous goods and the like containing them have the same health maintenance-promoting effect and beauty effect as raw royal jelly, and Since the risk of developing allergies is reduced compared to raw royal jelly and existing refined royal jelly, it can be used with peace of mind. In addition, the royal jelly fraction has a feature that even when blended with a liquid composition such as a beverage, turbidity and precipitation are less likely to occur compared to raw royal jelly.

  The raw royal jelly used as a raw material for preparing the royal jelly fraction of the present invention is not particularly limited in the type of bee secreted, its production area, and the supply form (raw or frozen). Examples of the bee species to be secreted include Apis mellifera, Apis cerana, Apis dorsata, and Apis florea. Production areas include Japan, South America, North America, Australia, China and other Asian regions, and Europe. Any of these royal jelly can be advantageously used as a raw material of the royal jelly fraction of the present invention, and it is more desirable to use a fresh royal jelly that is as fresh as possible or cryopreserved.

  For the preparation of the royal jelly fraction of the present invention, raw royal jelly as a raw material may be used as it is, but when performing ultramembrane filtration or gel filtration, the solubility of hydroxydecenoic acid, the filtration efficiency, etc. From the point of view, it is desirable to dilute before use. The dilution factor is not limited as long as the desired effect of the present invention can be achieved, and it is usually sufficient to dilute with an equal amount or more, preferably 2 times or more, particularly preferably 4 times or more of water. The water used for dilution is not particularly limited, and if necessary, for example, any of ultrapure water, ion exchange water, distilled water, magnetized water, cluster water, mineral water, tap water, and deep ocean water is advantageous. Available. The aqueous solution obtained by suspending and dissolving raw royal jelly in these waters can be used as it is for a purification step such as membrane filtration for reducing the polymer fraction described below. In addition, an alkaline agent or the like is added to fresh royal jelly, and the pH is usually 5.0 or higher, preferably 5.5 or higher, more preferably 5.5 to 12.0, and even more preferably 6.0 to 9.5. In particular, it is also possible to advantageously carry out purification by improving the solubility of hydroxydecenoic acid in the royal jelly fraction by adjusting to the range of 6.0 to 8.5. Since this pH is maintained substantially the same after the membrane filtration step described later, when the pH is lower than 5.0, for example, the recovery rate of hydroxydecenoic acid at the time of membrane filtration decreases, When diluted with cold water, turbidity and precipitation may occur. Further, when the pH is 6.0 or more, the recovery rate of hydroxydecenoic acid is stabilized, but the coloring degree (brownness) of the royal jelly fraction increases in proportion to the increase in pH. It is desirable to adjust the pH to 12.0 or less, more preferably 9.5 or less, considering the resistance of the filtration membrane, etc. .5 or less is particularly desirable. In the preparation of the royal jelly fraction of the present invention, either the dilution with water or the alkali treatment may be performed first. Usually, from the viewpoint of operability and pH retention, the dilution should be performed first. It is better to do it. It is also optional to dilute with alkaline water if necessary. Separately from this, the pH may be adjusted during the purification step or after the purification, if necessary.

  If the alkaline agent (pH adjuster) used for adjusting the pH of the raw royal jelly is a component that can be used in the production of the composition used in the application field of the fraction, such as food and drink, cosmetics, There is no particular limitation. In order to stably maintain the pH of the solution, it is optional to use a buffer that can be used in the field where the composition is used. Specifically, for example, examples of the alkali agent include sodium hydroxide, potassium hydroxide, calcium hydroxide and the like, and in the case of cosmetics, ethanolamines and the like may be used in addition to the above. it can. In addition, it is optional to use, for example, a phosphate buffer or an acetate buffer as the buffer. The addition of these alkali agents and buffering agents may be performed at once, may be performed in several times, or may be performed alternately with dilution.

  There are no particular restrictions on the amount of the alkaline agent or buffer used for adjusting the pH in the royal jelly fraction of the present invention as long as the desired pH can be maintained, but when used as a food or drink. It is desirable to use it in a range that does not affect the physical properties when it is blended into the composition. Specifically, when the pH is measured by diluting with deionized water or dissolving in deionized water so that the hydroxydecenoic acid contained in the royal jelly fraction of the present invention is 0.05 mg / mL, the pH is 5.0 or more, desirably 5.5 or more, more desirably 5.5 to 12.0, more desirably 6.0 to 9.5, and particularly desirably 6.0 to 8.5. desirable. The royal jelly fraction prepared from the raw royal jelly whose pH is adjusted under such conditions is an ingredient that contributes to the useful physiological activity of royal jelly as compared to the case where the allergenicity is reduced and the pH is not adjusted. Since the solubility of hydroxydecenoic acid in water is improved, the fraction can be dissolved in deionized water at 25 ° C. so that the hydroxydecenoic acid concentration is 5 mg / mL and cooled to 4 ° C. It has the characteristics that turbidity and precipitation that occur in the case of raw royal jelly and refined royal jelly are less likely to occur, and the clarity of the solution is also improved.

  The purification method for preparing the royal jelly fraction of the present invention has allergenicity without substantially losing the useful physiological activity inherent in the royal jelly (hereinafter sometimes simply referred to as “useful physiological activity”). Any method can be used as long as it can be removed. Specifically, any method can be used as long as the amount of protein can be reduced to a desired amount or less. Specific purification methods include, for example, water dilution, centrifugation, membrane filtration, filtration, concentration, fractional precipitation, salting out, dialysis, ion exchange chromatography, gel filtration chromatography, adsorption chromatography, isoelectric point chromatography. In addition, one or more methods commonly used in the art for purifying protein-containing substances such as hydrophobic chromatography, reverse phase chromatography, affinity chromatography, gel electrophoresis, and isoelectric focusing can be used as appropriate. In particular, fractionation with an ultrafiltration membrane is particularly preferred because it can not only completely remove the high molecular fraction, but also has almost no loss of a low molecular fraction having useful physiological activity. The useful physiological activity of royal jelly as used in the present invention refers to the maintenance / promotion of health represented by the royal jelly's collagen production enhancing action, IL-4 production inhibitory action, testicular function activating action, etc. It means all the physiological activities useful for humans and animals involved in the suppression of inflammation, immune enhancement action, cell activation action, etc., other than the above, for example, TNF-α production inhibition action, IgE production inhibition action, Anti-atopic action, contact hypersensitivity inhibiting action, life extending action, nourishing and tonic action, anti-fatigue action, etc. can be exemplified, and the royal jelly fraction of the present invention has one or more of these useful physiological activities. What is necessary is just to comprise, 2 or more types are desirable, and 3 or more types are especially desirable.

  As the pore size of the ultrafiltration membrane suitable for use in the removal of the polymer fraction of the above-mentioned raw royal jelly water dilution or the pH-adjusted water dilution, the polymer fraction is preferably obtained by membrane separation. Is a fraction having a molecular weight of 30,000 daltons or more, more preferably a fraction having a molecular weight of 10,000 daltons or more, and particularly preferably a fraction having a molecular weight of 6,000 daltons or more. There is no particular limitation. For example, the molecular weight cut off is 1,000 to 30,000 daltons, preferably 2,000 to 10,000 daltons, and more preferably the molecular weight cut off is selected from 2,000 to 6,000 daltons. It is preferred to use a filtration membrane. When the molecular weight cut off of the ultrafiltration membrane is less than 2,000 daltons, there is a concern that even useful low molecular weight substances may be removed, and in the case of a membrane having a fractional molecular weight larger than 30,000 daltons, the filtrate has allergenicity The allergenicity may increase due to an increase in the protein contamination rate. This polymer fraction can be removed by gel filtration chromatography or the like, and such a method may be used for removing the polymer fraction for preparing the royal jelly fraction of the present invention. In view of work efficiency and industrial applicability, it is preferable to use an ultrafiltration membrane.

  The fraction obtained by removing the polymer obtained by ultrafiltration or the like may be used as it is as the royal jelly fraction of the present invention. Usually, the low molecular weight substance obtained by ultrafiltration or gel filtration is often diluted considerably than the original concentration in the raw material, and in such a case, it can be concentrated and used as necessary. . Concentration methods such as heat concentration and reduced pressure concentration (including freeze drying) are appropriately employed as necessary. Among these, vacuum concentration that can be performed at a relatively low temperature is preferable because there is little concern about heat denaturation of components involved in useful physiological activity contained in royal jelly.

  The royal jelly fraction of the present invention obtained by the above method retains hydroxydecenoic acid contained in raw royal jelly without substantial loss, and is derived from royal jelly with respect to 1 part by mass of hydroxydecenoic acid. A fraction containing less than 0.05 parts by weight of the protein, and having physiological activities such as collagen production enhancing action and IL-4 production inhibitory action of royal jelly, and 45 μg of hydroxydecenoic acid. When the serum of a mouse immunized with an amount of the royal jelly fraction contained is determined by the passive skin anaphylaxis method using rats, allergenicity to the royal jelly fraction is not detected.

  The royal jelly fraction of the present invention reduces the protein that is a polymer substance that causes allergenicity, and its physicochemical index includes the mass ratio of hydroxydecenoic acid and protein, The royal jelly fraction of the present invention preferably has a ratio of protein less than 1: 0.05, and more preferably 1: 0.02 or less of protein. In the case of raw royal jelly, the mass ratio of hydroxydecenoic acid to protein is usually in the range of 1: 2 to 10, and in the existing purified royal jelly described in WO 2004/21803, 1: 0.3 to In the range of 1, the royal jelly fraction of the present invention is further purified and can be clearly distinguished from it. In addition, when the mass ratio of hydroxydecenoic acid and protein becomes 1: 0.05 or more and the ratio of the protein increases, depending on the amount of intake, the fraction or a composition containing the same , As a royal jelly, may cause allergies if taken orally in the same amount as normal.

  The amount of hydroxydecenoic acid contained in the royal jelly fraction of the present invention is usually 1 mg / g considering that the royal jelly fraction may be ingested as it is and that hydroxydecenoic acid has useful physiological activity. The above is desirable, 5 mg / g or more is more desirable, and 20 mg / g or more is particularly desirable. When the amount of hydroxydecenoic acid is less than 1 mg / g, the amount of intake for oral intake of hydroxydecenoic acid as a royal jelly may be too high, which may hinder use. .

  The protein in the present invention is quantified by Bradford method using human or bovine serum albumin as a standard protein, and the amount of nitrogen is measured by alkaline potassium peroxodisulfate decomposition-ultraviolet spectrophotometry. In addition, useful physiological activity, amount of hydroxydecenoic acid and allergenicity possessed by royal jelly can be measured and confirmed by the measurement methods shown in the following experimental examples. In addition, in the present specification, when referred to as raw royal jelly conversion, it means that the amount of the corresponding existing refined royal jelly and royal jelly fraction is converted to the mass of the raw royal jelly as the starting material required for the production thereof. If the amount is 1 g of raw royal jelly (total mass including water) as a starting material and purified, this is expressed as 1 g in terms of raw royal jelly.

  In the royal jelly fraction of the present invention, since the amount of high molecular protein is reduced, the amount of nitrogen in the solid content is reduced, and the mass ratio of hydroxydecenoic acid to nitrogen is 1: 0.3 or more. Since the content ratio of nitrogen is low, raw royal jelly having a mass ratio of hydroxydecenoic acid to nitrogen of approximately 1: 1, royal jelly obtained by enzymatic treatment thereof, and existing refined royal jelly having a mass ratio of 1: 0.5 or more It can be easily distinguished. In addition, the royal jelly fraction of the present invention may be abbreviated as “CBB” when the royal jelly fraction containing 30 μg of hydroxydecenoic acid is subjected to SDS-PAGE. ) Protein bands are not detected by visual observation with staining.

  The royal jelly fraction of the present invention having the above-described features is useful for physiological physiology such as collagen production enhancing action, anti-inflammatory action, cell activation action represented by testicular function activation action, life extension action, etc. inherent in royal jelly. Since the activity is substantially retained and the whitening effect is enhanced, food and drinks including health supplements for the purpose of maintaining and promoting health, whitening, beauty, skin, anti-wrinkle, anti-aging, etc. It can be advantageously used as an external preparation for skin such as cosmetics, quasi-drugs, pharmaceuticals, and skin miscellaneous goods.

  The royal jelly fraction of the present invention is useful by itself, but can also be advantageously used as a form of a composition formed by blending with other components. The present invention also provides such a composition. The composition according to the present invention usually comprises one or more components that are orally or percutaneously applied to mammals including humans or externally acceptable with the royal jelly fraction, for example, It can be advantageously used in the fields of foods, beverages, feeds / feeds / pet foods, cosmetics, and other external skin preparations, and pharmaceuticals. As an ingredient acceptable for oral or transdermal application to mammals including human being used in the present invention, those usually used in individual fields of application of the composition of the present invention, such as water, alcohol, starch, Examples include proteins, amino acids, fibers, carbohydrates, lipids, fatty acids, vitamins, minerals, flavoring agents, coloring agents, sweeteners, seasonings, spices, preservatives, emulsifiers, surfactants, and physiologically active substances. There is no restriction | limiting in particular in the form of the composition of this invention containing the above components, It provides with desired forms, such as a powder, a granule, a tablet, a paste, an emulsion, a cream, a solution.

  The composition formed by blending the royal jelly fraction of the present invention can be made into a solid form by blending the royal jelly fraction and cyclodextrins, anhydrous carbohydrates or dextrins and drying. Further, powders, granules, tablets and the like can be used. Examples of cyclodextrins include α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin. Examples of anhydrous sugars include anhydrous α, α-trehalose, anhydrous maltose, and anhydrous cyclic tetrasaccharide. In particular, anhydrous α, α-trehalose can be easily prepared from commercially available trehalose 2-hydrated crystals (for example, Hayashibara Shoji Co., Ltd., trade name “Treha”) by the method disclosed in Japanese Patent No. 3168550. This is desirable because it can be used. Also, as anhydrous maltose, commercially available anhydrous crystalline maltose (for example, Hayashibara Shoji Co., Ltd., trade name “Fine Tose”) can be advantageously used. Furthermore, cyclo {→ 6) -α-D-glucopyranosyl- (1 → 3) -α-D-glucopyranosyl- (1 → 6) -α-D-glucopyranosyl- disclosed in International Publication WO 02/10361, etc. Cyclotetrasaccharide having a structure of (1 → 3) -α-D-glucopyranosyl- (1 →} (cyclonigerosylnigerose), a cyclo described in JP-A No. 2005-95148 and the like {→ 6) -Α-D-glucopyranosyl- (1 → 4) -α-D-glucopyranosyl- (1 → 6) -α-D-glucopyranosyl- (1 → 4) -α-D-glucopyranosyl- (1 →} Cyclic tetrasaccharide having (cyclomaltosyl maltose), disclosed in JP-A-2005-95148 Cyclic pentasaccharide having the structure of cyclo {→ 6)-[α-D-glucopyranosyl- (1 → 4)] n-α-D-glucopyranosyl- (1 →} (n means 4 or 5) It is also optional to use anhydrous cyclic saccharides such as glycans and cyclic hexasaccharides, among which non-reducing saccharides and dextrins are preferred because they are less likely to cause browning reactions with royal jelly-derived components, especially α, α -Trehalose, cyclic tetrasaccharides and their saccharide derivatives also have a function of enhancing the physiological activity of the royal jelly fraction composition, and are therefore preferably used in combination from this point of view, and as dextrin, relatively low DE (dextrose Equivalent) is desirable because it has a strong effect of suppressing the browning of the royal jelly fraction. In consideration of the solubility of dextrin itself in water, D Of about 6.1 to 16.7 is used, and preferably about 7.1 to 11.5, in particular starch disclosed by the same applicant in the international patent application number PCT / JP2008 / 57879. Α-Glucosyltransferases produced by Bacillus circulans PP710 (Industrial Administrative Research Institute Microbial Deposit Center, Deposit No. FERM BP-1077) and the like are allowed to act on the partially decomposed product to produce α-1, Multi-branched glucans having an increased occupancy ratio of a branched structure composed of bonds other than α-1,4 bonds such as 6 bonds and α-1,3 bonds are particularly desirable. α-glucan, and 2,3,6-trimethyl-1,4,5-triacetylglucito in methylation analysis And the ratio of 2,3,4-trimethyl-1,5,6-triacetylglucitol in the range of 1: 0.6 to 1: 4, and 2,3,6-trimethyl-1,4, The total of 5-triacetylglucitol and 2,3,4-trimethyl-1,5,6-triacetylglucitol accounts for 60% or more of the partially methylated glucitol acetate, -Trimethyl-1,3,5-triacetylglucitol is 0.5% or more and less than 10% of partially methylated glucitol acetate, and 2,4-dimethyl-1,3,5,6-tetra A branched α-glucan in which acetyl glucitol has a structure of 0.5% or more of partially methylated glucitol acetate is particularly desirable because it is rich in dietary fiber components and has high water solubility.

  A composition obtained by blending the royal jelly fraction of the present invention with an anhydrous saccharide is, for example, a mixture of the royal jelly fraction and an anhydrous saccharide, and further mixing other components as necessary. The mixture can be obtained by dehydration drying or, if necessary, by subjecting it to a normal drying step such as reduced pressure drying, vacuum drying, freeze drying, fluidized drying, heat drying or the like. More specifically, when an anhydrous saccharide is used, when the anhydrous saccharide has the property of taking water and converting it into a water-containing crystal, the crystalline or non-crystalline anhydrous saccharide is converted into a royal jelly fraction. The amount of the fraction (including water) is usually mixed 4 times or more, more preferably 8 times or more, and further mixed with other components as necessary. The solid royal jelly fraction may be prepared by drying at 40 ° C. or lower, usually 4 hours or longer, preferably 8 hours or longer, more preferably 12 hours or longer, followed by dehydration and drying. In addition, the above-mentioned sugar, its hydrate or dextrin is usually added to the royal jelly fraction of the present invention in an equivalent amount or more, preferably twice as much as the amount of the solid content of the royal jelly fraction. Mix the above and prepare a solid royal jelly fraction by a method such as vacuum drying, vacuum drying, freeze drying, fluid drying, spray drying, etc. at 60 ° C. or lower, desirably 40 ° C. or lower, more desirably 30 ° C. or lower. It is also optional to do. The royal jelly fraction in a solid form thus prepared is further subjected to a drying step, if necessary, and then, using a pulverizer, a classifier, a granulator, a tableting machine, etc. It is also possible to advantageously carry out a desired form such as a granule or a tablet, and further, for example, fill the capsule with the powder or the granule as necessary.

  Commercially available glycosylated vitamin C (L-ascorbic acid 2-glucoside) can be further blended with the composition of the present invention as described above as necessary. Although it is well known that L-ascorbic acid has an action of enhancing collagen production in vivo, L-ascorbic acid has a drawback that it is unstable and susceptible to oxidative degradation. On the other hand, glycosylated vitamin C is a chemically stable substance and is first decomposed in vivo to release L-ascorbic acid, effectively enhancing the useful physiological activity of the royal jelly fraction. Can do. The timing of blending this glycosylated vitamin C into the composition of the present invention may be blended from the blending of raw materials to the completion of the product or into the finished product.

  Further, when an antioxidant component is further added to the composition of the present invention as required, further stabilization of the component involved in useful physiological activity contained in the royal jelly fraction can be achieved, and These physiological activities can be enhanced additively or synergistically. Therefore, according to the application object or application area of the composition, for example, it can be advantageously carried out when the composition is transported by a ship or the like without temperature control or used in a high temperature area. Although the antioxidant used in the present invention is not limited to a specific type, when the composition is used as an edible for mammals including humans, it is desirable to select appropriately from those normally used in the food field. . Specific examples of antioxidant components commonly used in the food field include flavonoids, polyphenols, vitamin E, carotene, vitamin C, and derivatives thereof. As flavonoids, more specifically, rutin, hesperidin, naringin, quercetin, and saccharides such as glucose or a polymer thereof bonded to each of them, glycosylated rutin, glycosylated hesperidin, glycosylated naringin, glycosyl transfer Examples include quercetin. More specifically, examples of the polyphenol include catechin, gallic acid, resveratrol and the like. Furthermore, plant extracts such as Enju extract, rosemary extract, eucalyptus extract, grape seed extract and the like can be advantageously used as an antioxidant component in the present invention. Although the content of these antioxidant components in the composition is not particularly limited, when the composition is used as an edible, in consideration of the influence on the taste, according to the blending ratio usually used in the food field, It is desirable to use it below or below. Moreover, when using for the purpose of strengthening these components, it is also optional to add the quantity which can anticipate the expected effect.

In addition to the above, Chinese parsley, paffia, guarana, indigo extract, perilla extract, propolis extract, kotarahim extract, pine extract, sasa extract, binic extract, rocca kurishi extract, collagen, hyaluronic acid, glucosamine, chondroitin, collagen, lacto Sucrose, coenzyme Q ₁₀ , α-lipoic acid, carnitine, pullulan, the same applicant disclosed in Japanese Patent Application No. 2008-107313 and International Publication No. WO2008 / 047758 (International Application No. PCT / JP2007 / 70077) -Combined use of components having physiological activity such as aminophenol and derivatives thereof is also possible.

  Desirable food forms of the composition of the present invention include, for example, ice cream, ice candy, ice confectionery such as sherbet, syrup such as ice honey, butter cream, custard cream, flower paste, peanut paste, fruit paste and the like Paste such as paste, chocolate, jelly, candy, gummy jelly, caramel, chewing gum, pudding, cream puff, sponge cake, etc.Processed fruit or processed vegetables such as jam, marmalade, syrup pickles, confectionery, manju, uirou, ann, mutton, water goat candy , Japanese sweets such as castella and candy, soy sauce, powdered soy sauce, miso, powdered miso, mayonnaise, dressing, vinegar, three cups of vinegar, table sugar, coffee sugar and the like. Desirable beverage forms include, for example, alcoholic beverages such as synthetic liquor, brewed liquor, fruit liquor, Western liquor, juice, mineral beverage, carbonated beverage, lactic acid beverage, lactic acid bacteria beverage, sports drink, drink agent, tea, tea, oolong tea, coffee And soft drinks such as cocoa.

  The royal jelly fraction of the present invention is used as it is or in the form of a composition containing other ingredients, for example, feed for domestic animals such as domestic animals, poultry, pets, other bees, moths, insects, fish, It can be advantageously used as a feed, pet food, and the like. As other components that can be blended in the composition having such a form, for example, grains, starches, sucrose, saccharides, oils and fats, seeds, beans, fish and shellfish, meat, which are usually used in each field , Eggs, milk, plant protein extracts, fruits, mushrooms, algae, vitamins, minerals, amino acids, yeast, pasture, bagasse, corn cob, rice straw, hay, oil meal, bran, soybean meal In addition, one or more of feed, feed, and pet food ingredients such as various fermented rice cakes may be mentioned. As a form of the composition, it can be advantageously used in a solid form such as powder, granule, tablet, paste, gummy, or liquid form such as emulsion, drink, soft drink. In addition, when an animal ingests the royal jelly or the composition, it may be ingested as it is, or it may be added to a feed or feed such as concentrated feed, roughage or pet food, or dissolved in drinking water. Is voluntary.

  Examples of desirable skin external compositions (including cosmetics, pharmaceuticals, and quasi drugs) of the composition of the present invention include aqueous solutions, solubilization systems, emulsification systems, powder dispersion systems, solid stick systems, and water-oils. The present invention can be applied in various dosage forms such as a two-layer system and a water-oil-powder three-layer system. In addition, there is no particular limitation on the specific form, such as basic cosmetics, cleaning cosmetics, bath cosmetics, hair cosmetics, tanning / sunscreen cosmetics, makeup cosmetics, oral cosmetics such as toothpastes, hair growth agents, It is applied to the skin, the outer skin such as the lips and the scalp, and the mouth as a hair restorer, soap and the like, and includes quasi-drugs, pharmaceuticals, and skin external goods having these forms. More specifically, ointments, creams including cleansing creams, emulsions, lotions, essences, jellies, gels, packs, shampoos, rinses, hair treatments, masks, mascaras, eyeliners, hair restorers, lipsticks (lipsticks), lips Gloss, foundation, blusher, eye shadow, powder, nail polish, soap, body soap, bath powder, toothpaste, moisturizing toothpaste, toothpaste, water toothpaste, medicated toothpaste, mouth freshener, mouth cooler, mouthwash, mouthwash Etc. can be illustrated.

  Moreover, it is possible to mix | blend a well-known component with the external composition for skin containing the royal jelly fraction of this invention according to the specific form. That is, in the case of the external preparation for skin, one or more of the components that can be used for the external preparation for skin and the components that can be used for the food are usually within the range that does not impair the intended effect of the present invention. Can be blended. For example, oil components, surfactants, preservatives (antibacterial agents), fragrances, whitening agents, moisturizers, thickeners, antioxidants, chelating agents, UV absorbers / scattering agents, vitamins, amino acids, anti-inflammatory agents , Blood circulation promoter, seaweed extract, astringent, anti-wrinkle agent, cell activator, anti-aging agent, hair growth / hair growth agent, transdermal absorption promoter, water, alcohols, water-soluble polymer, pH adjuster , Foaming agents, powders, pharmaceuticals, quasi-drugs, cosmetics, food additives, active ingredients for pharmaceuticals, quasi-drugs, etc. May be manufactured.

  Specifically, as oil components, macadamia nut oil, castor oil, olive oil, cacao oil, coconut oil, palm oil, tree wax, jojoba oil, grape seed oil, avocado oil and other vegetable oils, mink oil, egg yolk oil, etc. Animal fats, waxes such as beeswax, whale wax, lanolin, carnauba wax, candelilla wax, hydrocarbons such as liquid paraffin, squalane, microcrystalline wax, ceresin wax, paraffin wax, petrolatum, capric acid, myristic acid Natural and synthetic fatty acids such as palmitic acid, stearic acid, behenic acid, lanolin fatty acid, linoleic acid, linolenic acid, lauric acid, myristic acid, oleic acid, isostearic acid, cetanol, stearyl alcohol, hexyl decanol, octyldodecanol, lauryl Alcoa And natural and synthetic higher alcohols such as capryl alcohol, myristyl alcohol, cetyl alcohol, cholesterol, phytosterol, and esters such as isopropyl myristate, isopropyl palmitate, octyldodecyl myristate, octyldodecyl oleate, cholesterol oleate, etc. be able to.

  Surfactants include sorbitan monolaurate, sorbitan monopalmitate, sorbitan sesquioleate, sorbitan trioleate, polyoxyethylene sorbitan monolaurate, polyoxethylene sorbitan monostearate, polyethylene glycol monooleate, polyethylene glycol alkylate, Nonionic surfactants such as polyoxyethylene alkyl ether, polyglycol diether, lauroyl diethanol amide, fatty acid isopropanol amide, maltitol hydroxy fatty acid ether, alkylated polysaccharide, alkyl glucoside, sugar ester, lipophilic glycerin monostearate Self-emulsifying glycerin monostearate, polyglycerin monostearate, polyglycerin alkylate, so Vitan monooleate, polyethylene glycol monostearate, polyoxyethylene sorbitan monooleate, polyoxyethylene cetyl ether, polyoxyethylenated sterol, polyoxyethylenated lanolin, polyoxyethylenated beeswax, polyoxyethylene hydrogenated castor oil, etc. Nonionic surfactant, sodium stearate, potassium palmitate, cetyl sulfate, sodium lauryl phosphate, triethanolamine palmitate, sodium polyoxyethylene lauryl phosphate, sodium N-acyl glutamate, sodium palmitate, sodium laurate, Sodium laurate, potassium lauryl sulfate, alkyl sulfate triethanolamine ether, funnel oil, linear dodecyl benzene sulfate, polio Anionic surfactants such as siethylene hydrogenated castor oil maleic acid, acylmethyltaurine, cationic surfactants such as stearyldimethylbenzylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium chloride, benzalkonium chloride, laurylamine oxide, alkyl hydrochloride Examples include amphoteric surfactants such as aminoethylglycine solution and lecithin.

  Preservatives (antibacterial agents) include benzoic acid and its salts, salicylic acid and its salts, sorbic acid and its salts, dehydroacetic acid and its salts, paraoxybenzoic acid alkyl esters and other paraoxybenzoic acid esters, 2, 4, 4'-trichloro-2'-hydroxydiphenyl ether, 3,4,4'-trichlorocarbanilide, hexachlorophene, benzalkonium chloride, phenoxyethanol, hinokitiol, resorcinol, ethanol, 1,3-butylene glycol, photosensitizer 201 Etc. can be illustrated.

  As fragrances, benzaldehyde, benzyl benzoate, phenylacetic acid, sandalol, eugenol, rillial, rilal, linalool, 2-methyl-3- (4-methylphenyl) -propanal, musk ketone, cinnamic aldehyde, belt fix, methyl ionone, Geranylformate, iso-E super, γ-undecalactone, hexyl salicylate, cis-3-hexenyl salicylate, methyl dihydrojasmonate, tetrahydrofurfuryl 3-mercaptopropionate, covanol, vanillin, vanillar, geranium oil, peni Examples include royal oil, birch oil, and alumois oil.

  Examples of whitening agents include L-ascorbic acid and derivatives thereof and salts thereof, alkoxysalicylic acids and salts thereof, hydroquinone derivatives such as hydroquinone and glycosides thereof, tranexamic acid and derivatives thereof and salts thereof, derivatives of resorcin, Examples include kojic acid and derivatives thereof and salts thereof, ellagic acid and linoleic acid and salts thereof, chamomile extract, tetrahydrocurcuminoid, cyanobacteria extract, 2-aminophenol and derivatives thereof.

  As the humectant, polyhydric alcohols such as erythritol, xylitol, sorbitol, maltitol, glycerin, propylene glycol, 1,3-butylene glycol, polyglycerin, polyethylene glycol, dipropylene glycol, 1,2-pentanediol, isoprene glycol, etc. , Glucose, maltose, trehalose, carbohydrate derivative of trehalose, dextrin, cyclodextrin, cyclo {→ 6) -α-D-glucopyranosyl- (1 → 3) -α- disclosed in WO 02/10361 pamphlet A cyclic tetrasaccharide having a structure of D-glucopyranosyl- (1 → 6) -α-D-glucopyranosyl- (1 → 3) -α-D-glucopyranosyl- (1 →} (cyclonigerosyl nigerose: Cyclonigerosyl) nigelose), cyclo {→ 6) -α-D-glucopyranosyl- (1 → 4) -α-D-glucopyranosyl- (1 → 6) -α-D-glucopyranosyl-((Japanese Patent Application Laid-Open No. 2005-95148) 1 → 4) -α-D-glucopyranosyl- (1 →} cyclic tetrasaccharide (cyclomaltosylmaltose) and other sugars, amino acids, NMF components such as sodium lactate and sodium pyrrolidonecarboxylate (natural moisturizing) Ingredients), glycogen, locust bean gum, xyloglucan, quince seed, carrageenan, pectin, mannan, curdlan, succinoglucan, galactan, dermatan sulfate, keratan sulfate, chondroitin, chondroitin sulfate, mucoitin sulfate, keratosulfate, chitin Mucopolysaccharides such as heparan sulfate and hyaluronic acid, hydrolysates of these mucopolysaccharides, water-soluble polymer substances such as proteins and peptides such as silk and collagen, and hydrolysates thereof, salts thereof, dimethylpolysiloxane, Examples include silicones such as methylphenylsiloxane, culture supernatants such as lactic acid bacteria and bifidobacteria, among others, trehalose, trehalose carbohydrate derivatives, cyclic tetrasaccharides, hyaluronic acid and chondroitin, which have a strong moisturizing effect. It is desirable to use one or two or more selected from sulfuric acid, its salts and hydrolysates.

  Thickeners include sodium alginate, xanthan gum, aluminum silicate, quince seed extract, gum arabic, hydroxyethyl guar gum, carboxymethyl guar gum, guar gum, dextran, tragacanth gum, starch, pullulan, chitin, chitosan, agar, cellulose, etc. Natural synthetic materials such as hydroxypropylcellulose, methylhydroxypropylcellulose, methylcellulose, carboxymethylcellulose, hydroxyethylcellulose, soluble starch, cationized cellulose, carboxymethylchitin, semi-synthetic polymer materials, carboxyvinyl polymer, polyvinyl alcohol, polyvinylpyrrolidone Examples thereof include synthetic polymer substances such as vinyl alcohol / vinyl acetate copolymer.

  Examples of the antioxidant include dibutylhydroxytoluene, butylhydroxyanisole, propyl gallate, L-ascorbic acid, vitamin E, catechins, flavonoids and derivatives thereof.

  Examples of chelating agents include disodium edetate, ethylenediaminetetraacetate, pyrophosphate, hexametaphosphate, citric acid, tartaric acid, gluconic acid, and the like.

  Examples of the pH adjuster include sodium hydroxide, triethanolamine, nitrilotriethanol, citric acid, sodium citrate, boric acid, borax, potassium hydrogen phosphate, and the like.

  Examples of ultraviolet absorbers / scatterers include paraaminobenzoic acid ultraviolet absorbers, anthranilic acid ultraviolet absorbers, salicylic acid ultraviolet absorbers, cinnamic acid ultraviolet absorbers, benzophenone ultraviolet absorbers, sugar ultraviolet absorbers, 3 -(4'-methylbenzylidene) -d-camphor, 3-benzylidene-d, 1-camphor, urocanic acid, urocanic acid ethyl ester, 2-phenyl-5-methylbenzoxazole, 2,2'-hydroxy-5 -Methylphenylbenzotriazole, 2- (2'-hydroxy-5'-t-octylphenyl) benzotriazole, 2- (2'-hydroxy-5'-methylphenylbenzotriazole), dibenzalazine, dianisoylmethane, 4-methoxy -4'-t-butyldibenzoylmethane, 5- (3,3-dimethyl-2 Norbornylidene) -3-pentan-2-one, 2-hydroxy-4-methoxybenzophenone, octyl dimethyl p-aminobenzoate, ethylhexyl p-methoxycinnamate Saina formate, can be exemplified titanium oxide, kaolin, talc and the like.

As vitamins, vitamin A and its derivatives, vitamin B ₁ and its derivatives, vitamin B ₂ and its derivatives, vitamin B ₆ hydrochloride, vitamin B ₆ tripalmitate, vitamin B ₆ dioctanoate, vitamin B ₁₂ , vitamin B ₁₅ And vitamin B such as derivatives thereof, L-ascorbic acid and derivatives thereof, α-tocopherol, β-tocopherol, γ-tocopherol, vitamin E acetates such as vitamin E acetate, vitamin D, vitamin H, pantothenic acid, panthetin, Examples include flavonoids such as vitamin F, vitamin K, and vitamin P and derivatives thereof, vitamin U, ferulic acid, γ-oryzanol, α-lipoic acid, orotic acid, coenzyme Q10, and derivatives thereof. It may be a salt of Of these, L-ascorbic acid and its derivatives are desirable because they have a strong effect on enhancing the physiological action of royal jelly fractions and on enhancing collagen production, which is strongly involved in the suppression of wrinkle formation on the skin. In terms of strength, a glycoside of L-ascorbic acid is desirable, and L-ascorbic acid-2-glucoside is particularly desirable.

  As amino acids, glycine, alanine, valine, leucine, isoleucine, serine, threonine, phenylalanine, tyrosine, asparagine, glutamine, taurine, tryptophan, cystine, cysteine, methionine, proline, hydroxyproline, aspartic acid, glutamic acid, arginine, histidine , Lysine, carnitine, and derivatives thereof, and salts thereof may be used.

  Anti-inflammatory agents include allantoin or its derivatives allantoin, allantoin acetyl-dl-methionine, allantoin chlorohydroxyaluminum, allantoin dihydroxyaluminum, allantoin polygalacturonic acid, glycyrrhetinic acid or its derivatives glycyrrhetic acid, glycyrrhizic acid, glycyrrhetic acid Allantoin, glyceryl retinoic acid, stearyl glycyrrhetinate, stearyl glycyrrhetinate stearate, disodium 3-succinyloxyglycyrrhetinate, dipotassium glycyrrhizinate, monoammonium glycyrrhizinate, pantothenic acid, pantothenyl alcohol, pantothenyl ethyl ether, acetyl pantothenyl ethyl ether , Benzoylpantothenyl ethyl ether, bread Calcium tenate, sodium pantothenate, acetyl pantothenyl ethyl ether, benzoic acid pantotenyl ethyl ether ester, pantothenic acid derivatives such as panthetin, vitamin E, d-δ-tocopherol, dl-α-tocopherol, dl-α-acetate L-ascorbic acid such as tocopherol, linoleic acid dl-α-tocopherol, nicotinic acid dl-α-tocopherol, vitamin E derivatives such as succinic acid dl-α-tocopherol, L-ascorbic acid, L-ascorbic acid-2-glucoside Glucosides, acylated derivatives of L-ascorbic acid glucoside, tetrahexyldecanoic acid ascorbic acid, ascorbic acid-tocopherol phosphate diester L-ass, in which L-ascorbic acid and tocopherol are bonded via a phosphate group Derivatives of L-ascorbic acid and / or their derivatives such as rubic acid sulfate ester, ascorbyl dipalmitate, ascorbyl palmitate, stearyl L-ascorbate, L-ascorbyl phosphate, ethyl L-ascorbate and acylated derivatives thereof Salts of alkali metals or alkaline earth metals, 2-aminophenol and derivatives thereof, pyridoxine hydrochloride, menthol, biotin, camphor, turpentine oil, zinc oxide, azulene, guaiazulene and derivatives thereof, mefenamic acid and derivatives thereof, phenylbutazone and Its derivatives, indomethacin and its derivatives, ibuprofen and its derivatives, ketoprofen and its derivatives, ε-aminocaproic acid, diclofenac sodium, diphenhydramine, tranexamic acid and its derivatives, Samethasone, cortisone and its esters, hydrocortisone and its esters, prednisone, prednisolone and other corticosteroids, antihistamines, cyanobacteria, asenya, acha, altea, arnica, aloe, ibutrano, nettle, age, echina, emium, turmeric, Ougon, Buckwheat, Barley, Hypericum, Orange, Valerian, Chamomile, Carrot, Achillea mugwort, Licorice, Cucumber, Goldfish, Gardenia, Kumazasa, Watercress, Gentian, Gentian pepper, Burdock, Comfrey, Salamander, Salamander, Salamander, Salamander , Perilla, Peonies, Birch, Sage, Hypericum perforatum, Kizota, Papilio spp, Achillea, Pepper Cucumber, Sembli, Soakhi, Taiso, Thyme, Chinese parsley, Capsicum, Toki, Tokinsenka, Tonin, Dokudami, Tolmentilla, Tadeai, Cha, Toki, Tokinsenka, Elderberry, Carrot, Parsley, Hakka, Sandalwood, Biwa, Butchabu Exemplifying ingredients such as grapes, broccoli, safflowers, buffalo, bodyfish, buttons, marooniers, mugwort, peaches, cornflowers, eucalyptus, mugwort, bayberry, lavender, roman chamomile, rosemary, etc. Can do.

  As seaweed extracts, there are extracts from brown algae, red algae, green algae, cyanobacteria, etc., specifically, kombu, macombu, wakame, hijiki, cypress, coral, palmaria, tsunomata, nori, aosa, anaaaosa, ascophyllum, Examples include extracts from Hibamata, Mozuku, Okinawa Mozuku, Himantaria and the like. This includes extracts from water plants such as sea cucumbers.

  In addition to the above, plant and animal ingredients such as propolis and herbs, ingredients such as extracts derived from them, minerals such as deep sea water, bitter and bitter ingredients, antibiotics, calcium hydrogen phosphate, organic Modified montmorillonite, silicic acid, anhydrous silicic acid, magnesium silicate, mica, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, bitumen, chromium oxide , Inorganic powders such as calamine, zeolite and carbon black, polyamide, polyester, polyethylene, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic acid resin, melamine resin, epoxy resin, polycarbonate Anthraquinone series, including fats, divinylbenzene / styrene copolymers, copolymers of two or more monomers of the above compounds, organic powders such as cellulose, safflower dyes, gardenia dyes, sicon dyes, cochineal dyes , Natural colorants such as anthocyanin, chalcone, and carotenoid pigments, photosensitive elements 101, 201, 201, 401, red 201, 202, red 204, red 205, red 220, red 226, red 228, red 405, orange 203, orange 204, yellow 204, yellow 401 and blue 404, etc., organic pigment powder, red 3 No., Red 104, Red 106, Red 227, Red 230, Red 401, Red 505, Orange 205, Yellow 4, Organic pigment powder such as Zirconium, Barium or Aluminum Lake of Color No. 5, Yellow No. 202, Yellow No. 203, Green No. 3 and Blue No. 1, natural colorants such as sicon dye and safflower dye, and organic synthetic dyes And coloring agents such as vitamins and derivatives thereof, powders loaded with functional components such as sugar-transferred glycyrrhizin and glycyrrhizin.

  In order to produce a composition comprising the royal jelly fraction of the present invention, the royal jelly fraction of the present invention can be used according to its purpose based on the components and individual contents as described above, that is, According to an appropriate composition selected according to the type of mammal to be used and its intake method, it may be contained in the process until the target composition is completed or in the finished product. Examples of the treatment method include dilution, concentration, drying, filtration, centrifugation, mixing, kneading, dissolution, melting, dispersion, suspension, emulsification, reverse micellization, penetration, crystallization, spraying, coating, adhesion, and spraying. A known processing method such as coating (coating), pouring, dipping, solidifying, carrying, or encapsulating may be appropriately selected, and further formed into a desired shape as necessary. The order of blending the components and the timing of various treatments may be any as long as they do not cause deterioration of the quality of the royal jelly fraction. For example, royal jelly immediately after preparation or stored at low temperature after preparation is available. What is necessary is just to perform a various process suitably as needed after using a fraction. In addition, in order to prevent deterioration of the quality of the royal jelly fraction in the production process, all the above steps are performed under the conditions of 60 ° C. or less, desirably 40 ° C. or less, and particularly desirably 30 ° C. or less. It is possible to prevent attenuation of useful physiological activity in The composition containing the royal jelly fraction of the present invention as described above is generally 0.0001% to 20%, preferably 0.001% of the royal jelly fraction in terms of solids per product mass. % To 10%. In addition, in the case of health supplements and the like for the purpose of taking royal jelly, it is optional to further increase the content in the composition.

  The composition according to the present invention as described above stably retains various useful physiological activities of raw royal jelly, so that the useful physiological activity is effectively exhibited in the living body used, and the resistance of the living body. Enhancement, early improvement of poor physical condition, maintenance / promotion of healthy condition, beauty, skin beautification, etc. In particular, it is also effective in enhancing testicular function in males for breeding animals. And since the allergenicity is reduced, the royal jelly fraction of this invention can be used in comfort. Therefore, the composition containing the royal jelly fraction of the present invention is extremely useful as a food for maintaining and promoting health, whitening and beauty, beverages, feeds, feeds, pet foods, skin external compositions and the like. .

  The amount and method of intake of the royal jelly fraction of the present invention or a composition comprising the fraction are not limited as long as the amount and method can exert the useful physiological activity of the royal jelly. Decenoic acid may be 0.001 to 100 mg, preferably 0.01 to 10 mg, particularly preferably 0.1 to 2 mg per kg body weight per day. Moreover, as an ingestion method, the required amount per day may be ingested at a time or may be ingested in several times. It may also be taken once every 2 to 3 days.

  Hereinafter, the present invention will be described in more detail based on experiments.

<Experiment 1: Effect 1 of ultrafiltration on allergenicity of royal jelly>
As described later, although it was confirmed that the existing purified royal jelly prepared by the method of International Publication No. WO2004 / 21803 is still weaker than raw royal jelly, allergenicity still remains. In order to further remove proteins that are considered to be the cause of allergy without reducing the components involved in the useful physiological activity, the following tests were conducted to examine the effect of ultrafiltration on the allergenicity of raw royal jelly. It was. That is, 50 g of fresh royal jelly (produced in Taiwan, containing 30 mg / g of hydroxydecenoic acid and 113 mg / g of protein) was suspended in 200 g of purified water. This water-diluted solution is subjected to ultrafiltration using an ultrafiltration membrane (Nippon Genetics Co., Ltd. or Advantech Toyo Co., Ltd.) having various molecular weights shown in Table 1, and the ultrafiltrate is recovered. Table 1 also shows the results of measuring the allergenicity of the test samples. In the evaluation of the allergenicity of the filtrate, these test samples (ultrafiltrate) all used an amount containing an equivalent amount of hydroxydecenoic acid. The presence or absence of allergenicity test, collagen production enhancing action and IL-4 production inhibitory action of each test sample was measured by the following method, and the results are also shown in Table 1.

<Method of allergenicity test>
In the allergenicity test, an antigenicity test using mice was carried out according to the method of Hirota et al. (Tadatoshi Hamada, Hiroshi Mizushima, “Ayumi of Medicine”, 100, 814, 1977). That is, using 5 female BALB / c mice (9 weeks old, sold by Nippon Charles River Co., Ltd.) per test group, a test sample containing 45 μg of each hydroxydecenoic acid 3 times every other week Immunization was performed by intraperitoneally administering 2.5 mg / animal of aluminum hydroxide gel (Alum, hereinafter referred to as “Alam adjuvant”) as an adjuvant. One week after the third immunization, blood was collected from the tail artery of each mouse, serum was collected using a microtenor, and subjected to PCA using rats. Specifically, 0.1 mL of each mouse serum diluted with physiological saline (10 times) was intradermally injected into the back of normal SD rats (11 weeks old, sold by Nippon Charles River Co., Ltd.) After 24 hours, 5 times the same amount as the test sample immunized to the mice was administered intravenously together with 1 mL of 0.5% Evans Blue dye solution.

<Method for determining the presence or absence of allergenicity>
The size of the blue spot observed at the mouse serum administration site was measured, and a sample showing a spot having an average diameter of 5 mm or more was determined as positive, and a sample showing a spot having an average diameter of less than 5 mm was determined as negative. The strength of allergenicity was evaluated in 6 stages, with 5 positive (+++++) through 5 negative in the test group and 5 negative (−).

<Method of measuring collagen production enhancing action>
The collagen production enhancing action was measured using a method for measuring collagen production using human fibroblasts in accordance with the method described in International Publication WO2005 / 034938. That is, human fibroblasts (sold by Kurashiki Boseki Co., Ltd., cell name “NHDF cells”) are 10% (v / v) fetal bovine serum and an appropriate amount of penicillin so that the cell concentration is 2.5 × 10 ⁴ cells / well. Dulbecco's modified Eagle's medium supplemented with Nikko (Nissui Sales Co., Ltd., hereinafter, Dulbecco's modified Eagle medium supplemented with fetal bovine serum and an appropriate amount of penicillin is abbreviated as "DMEM medium") (pH 7.1 to 7.4). ) And a 96-well microplate (available from Becton Dickinson) and cultured at 37 ° C. for 24 hours under 5% (v / v) CO ₂ conditions. Thereafter, the culture supernatant was removed, and each test sample was replaced with 200 μL of DMEM medium containing 50 μM L-ascorbic acid added so that the hydroxydecenoic acid concentration was 100 μg / mL, and cultured for 5 days. Thereafter, the amount of collagen produced by the cells per well in the microplate was quantified using a Sarkor Collagen Assay Kit (sold by Biocolor). For the measurement, 3 wells were used for each test sample.

<Method for determining presence or absence of collagen production enhancing action>
The determination of the presence or absence of a collagen production enhancing action is active when the amount of collagen is increased by 20% or more compared to when the cells are cultured only in a DMEM medium containing 50 μM L-ascorbic acid (◯). Alternatively, the case where there was an increase of less than 20% was regarded as having no activity (x).

<Method for measuring IL-4 production inhibitory action>
The measurement of IL-4 production inhibitory effect was carried out by the IL-4 production system using BALB / c mouse spleen cells immunized with ovalbumin containing alum adjuvant described in Experiment 2 of International Publication WO2004 / 019971 Made using. Specifically, spleen cells were collected from BALB / c mice immunized three times with Alum adjuvant using OVA as an antigen and suspended in a medium at 5 × 10 ⁶ cells / mL. 100 μL of this cell suspension was seeded on a 96-well microplate preliminarily immobilized with anti-CD3 antibody (5 μg / mL). Next, the test sample was diluted with phosphate buffered saline so that the hydroxydecenoic acid concentration was 400 μg / mL, and 50 μL of this was added. Further, 50 μL of medium was added to each well to make a total liquid volume of 200 μL. After culturing for 40 hours, the culture supernatant was collected, and the amount of IL-4 was measured by a solid-phase enzyme immunoassay (ELISA method). A control was prepared by adding phosphate buffered saline instead of the test sample and culturing in the same manner.

<Method for Determining IL-4 Production Inhibitory Effect>
The determination of the presence or absence of IL-4 production inhibitory activity is active when the production amount of IL-4 is suppressed by 20% or more compared to the control (◯), or when only less than 20% is suppressed Was not active (x).

  As is apparent from Table 1, the allergenicity of the test sample (ultrafiltrate) decreases and the molecular weight cut-off is 30,000 in proportion to the decrease in the molecular weight cut-off of the membrane used for ultrafiltration. The allergenicity was not observed in the test samples which were markedly reduced below dalton and fractionated with an ultrafiltration membrane of 10,000 dalton or less. Collagen production enhancing action was also confirmed in the case of test samples using ultrafiltration membranes of any fractional molecular weight. This result shows that a high molecular weight fraction is removed by fractionating a dilute solution of raw royal jelly with an ultrafiltration membrane having a molecular weight cut-off of 30,000 daltons or less, preferably 10,000 daltons or less. This indicates that a royal jelly fraction with reduced allergenicity can be prepared while maintaining the physiological activity of royal jelly. Further, considering the yield of low-molecular components involved in collagen enhancing action and the ease of processing, the ultrafiltration membrane used for the preparation of the royal jelly fraction of the present invention has a molecular weight cutoff of 2,000. It was determined that the use of a material selected from 1 to 10,000 daltons was more desirable, and that the material selected from 2,000 to 6,000 daltons was particularly desirable. It should be noted that the serum of mice immunized with a test sample fractionated with an ultrafiltration membrane having a molecular weight cut-off of 30,000 Dalton or less showed no PCA reaction when diluted 200-fold in any case. .

<Experiment 2: Effect 2 of ultrafiltration on allergenicity of royal jelly>
Experiment 1 confirms that allergenicity is effectively reduced by removing royal jelly with an ultrafiltration membrane selected from a molecular weight cut off of 2,000 to 10,000 daltons. A test for investigating the characteristics of the royal jelly fraction was performed as follows. That is, 500 g of fresh royal jelly (produced in Taiwan, containing 30 mg / g of hydroxydecenoic acid and 113 mg / g of protein) was suspended in 2,000 g of purified water, and ultrafiltration membrane (sales brand name “Microser AIP1013” sold by Asahi Kasei Corporation). And subjected to ultrafiltration using a molecular weight cutoff of 6,000). Ultrafiltration is conducted by adding purified water to an ultrafiltration concentrate (inner liquid: diluted raw jelly water) and collecting 4,000 mL of ultrafiltrate (filtrate) to prepare test sample 1. did. Moreover, based on the method of Experimental Example 3 of International Publication WO2004 / 21803, 60 g of the above-mentioned raw royal jelly was used as a raw material, and 240 g of deionized water at 4 ° C. was added thereto and stirred to be uniformly dispersed. The suspension was then centrifuged (5,000 × g, 10 minutes) and separated into a supernatant and a precipitate. 240 g of deionized water at 4 ° C. was again added to the resulting precipitate and treated in the same manner to obtain a precipitate. The first and second supernatants of the water fraction were combined into a water supernatant and subjected to ultrafiltration using the ultrafiltration membrane. The mixture was pushed with a small amount of water, and separated into an ultrafiltrate (filtrate) and an ultrafiltrate concentrate (inner liquid). The filtrate obtained by ultrafiltration was concentrated twice with an evaporator, and finally, an existing purified royal jelly was prepared together with the above-mentioned precipitate to obtain Test Sample 2. Further, 48 g of deionized water was added to 12 g of the raw royal jelly to make 60 g, and the mixture was stirred and dispersed in the same manner as above to prepare a raw royal jelly water diluted solution. Measures the mass of hydroxydecenoic acid, the mass of protein, and the mass of nitrogen, which are specific to royal jelly as an indicator of the components involved in the useful physiological activities of these test samples 1 to 3, and are said to be involved in sugar metabolism and prevention of aging Table 2 shows the results of SDS-PAGE as an index of the mass ratio of protein and hydroxydecenoic acid, the mass ratio of hydroxydecenoic acid and nitrogen, and the protein mass. Further, Table 2 shows the results of measuring the presence or absence of allergenicity, collagen production enhancing action, and IL-4 production inhibiting action of these test samples in the same manner as in Experiment 1. The protein mass was measured by Bradford method using human serum albumin as a standard protein, and the relative mass (%) of the protein mass contained in each sample was determined with the protein mass contained in the first raw royal jelly as 100. The amount of protein (%) is shown in Table 2. Hydroxydecenoic acid is measured by the HPLC method under the following conditions, and quantified by a calibration curve prepared using a hydroxydecenoic acid standard solution (20 μg / mL, 50 μg / mL, 100 μg / mL) prepared in advance. The relative amount (%) of the amount of hydroxydecenoic acid contained in each sample was determined with the amount of hydroxydecenoic acid contained in the raw royal jelly of 100 being shown in Table 2 as the amount of hydroxydecenoic acid (%). The nitrogen mass was determined by alkaline potassium peroxodisulfate decomposition-ultraviolet absorptiometry. For SDS-PAGE, each test sample containing 30 μg of hydroxydecenoic acid was converted into a commercially available gel for SDS-PAGE (sold by Daiichi Chemical Co., Ltd., trade name “PAG Mini“ Daiichi ”) 10/20. )), And after electrophoresis by a conventional method, stained with Coomassie Brilliant Blue (CBB), and the presence or absence of a stained band is confirmed by visual observation, and a plurality of bands are clearly confirmed (++++). The band of the royal jelly major protein 1 near 52.8 kilodalton is clearly confirmed (++++), and only the band of the royal jelly major protein 1 near 52.8 kilodalton is clearly confirmed. (++), but only a band of royal jelly major protein 1 near 52.8 kilodaltons is confirmed although it is thin (+). No (-) was evaluated in five stages of.

<HPLC analysis conditions for hydroxydecenoic acid>
Standard solution: Diluted commercially available hydroxydecenoic acid (sold by Alfresa Pharma Co., Ltd.) Column: Reversed phase column (YMC-Pack AQ303-ODS)
(Φ4.6mm × 250mm, YMC Co., Ltd. sales))
Column temperature: 40 ° C
Detector: UV210nm Mobile phase: 60% methanol (pH adjusted to 2.2 with phosphoric acid)
Flow rate: 0.7 mL / min, sample volume: 20 μL

  As is clear from Table 2, the amount of hydroxydecenoic acid recovered in test sample 1 (royal jelly fraction) was about 70% compared to test sample 3 (raw royal jelly water diluted solution) used as the starting material for purification. The protein amount was reduced to 0.03%. On the other hand, in test sample 2 (existing purified royal jelly), the amount of hydroxydecenoic acid is reduced to about 60% and the protein amount is up to 8% compared to test sample 3 (raw royal jelly water diluted solution). Diminished. The results of SDS-PAGE correlate with the measurement results of protein mass. In test sample 2 (existing purified royal jelly), the protein band was determined to be (+), and the band of royal jelly major protein 1 was around 52.8 kilodaltons. Only a few were confirmed. In test sample 1 (royal jelly fraction), no protein band was observed, and it was determined as (−). The mass ratio of hydroxydecenoic acid to nitrogen was 1: 0.15 in Test Sample 1, whereas it was 1: 0.56 in Test Sample 2, and 1: 0.91 in Test Sample 3. In test sample 1, allergenicity was not recognized and it was determined to be minus (−), whereas test sample 2 was determined to be 3 plus (++++), and test sample 3 was determined to be 5 plus (++++). Collagen production enhancing action and IL-4 production inhibitory action were confirmed in any test samples.

  These results show that in test sample 1, allergenicity is not observed, and the amount of hydroxydecenoic acid, which is a component involved in the useful physiological activity of raw royal jelly, is not significantly reduced, and the amount of protein is reduced. It is shown that. In addition, the mass ratio of nitrogen to hydroxydecenoic acid decreased in proportion to the decrease in protein mass in the test sample, and was the lowest in test sample 1. In addition, the ultrafiltrate (test sample 1: royal jelly fraction) obtained by filtering a raw royal jelly water diluted solution through an ultrafiltration membrane having a molecular weight cut off of 6,000 has a collagen production enhancing action and an IL-4 production inhibiting action. Therefore, it was determined that the biological activity of raw royal jelly was retained.

<Experiment 3: solubility and coloring of hydroxydecenoic acid in pH-adjusted royal jelly fraction>

<Effect of pH on solubility of hydroxydecenoic acid>
A test for examining the effect of pH was conducted as follows using hydroxydecenoic acid, which is one of organic acids contained in royal jelly, as an index. That is, test sample 1 (royal jelly fraction) used in Experiment 2 was freeze-dried by a conventional method to prepare a powder product. After adding an excessive amount of this powdered product (the amount in which hydroxydecenoic acid is not completely dissolved even after pH treatment by dissolving in water) to a fixed amount of deionized water (25 ° C.), stirring and dissolving with a stirrer for 30 minutes , 6N (N) sodium hydroxide solution was added, the pH was adjusted to the values shown in Table 3, and the mixture was further stirred with a stirrer for 30 minutes, centrifuged at 15,000 rpm for 30 minutes, and the supernatant was recovered. After measuring the pH, the amount of hydroxydecenoic acid in the supernatant was measured, and the solubility of hydroxydecenoic acid in the royal jelly fraction is also shown in Table 3. Moreover, the result of having confirmed the presence or absence of coloring (degree of browning) of the collect | recovered supernatant with the naked eye is combined with Table 3, and is shown. The amount of hydroxydecenoic acid was measured by the same method as in Experiment 1. As for the degree of coloring of the supernatant, put it in a glass bottle with a cylindrical cap with a diameter of 12 mm, and by visual observation from the side, there is no coloring (-), light coloring is observed (+), and the browning of the solution is clear (++) and coloring was remarkable (++++). Further, the sample (recovered supernatant) having a pH of 5.0 or higher used for measuring the solubility of hydroxydecenoic acid (recovered supernatant) was heated to 25 ° C. so that the concentration of hydroxydecenoic acid was 5 mg / mL. Each of these was diluted with deionized water at 0 ° C. and placed in a glass bottle with a cap made of glass. After standing at 2 ° C. for 2 hours in a cooled state, the presence or absence of precipitation was confirmed by visual observation. . The results are also shown in Table 3. Moreover, the presence or absence of the allergenicity and collagen production enhancing action was measured by the same method as in Experiment 1, and the results are also shown in Table 3.

  As is apparent from Table 3, the solubility of hydroxydecenoic acid in the royal jelly fraction at 25 ° C. increases at pH 5.0 or higher, rapidly increases at pH 5.5 or higher, and significantly increases at pH 6.0 or higher. Admitted. The royal jelly fraction was colored at pH 9.0 or higher. The degree of coloration was determined to be ++ at pH 10.0 to 12.0 for the royal jelly fraction, and ++ at pH 13.0. In addition, the occurrence of precipitation when the solution was cooled was suppressed when the pH of the test sample was adjusted to 5.0 or higher, preferably 5.5 or higher, and the effect became remarkable at pH 6.0 or higher. As a result, the royal jelly fraction can significantly increase the solubility of hydroxydecenoic acid by setting its pH to 5.0 or higher, preferably 5.5 or higher, more preferably 6.0 or higher. It tells you what you can do. In addition, from the viewpoint of coloring, coloring can be clearly confirmed at pH 10.0 to 12.0, and is particularly noticeable at pH 13.0, so that it can be used for the preparation of white and colorless compositions. Determined that the pH of the royal jelly fraction was preferably 5.0 to 9.5, and particularly preferably 6.0 to 8.5. Except for coloring, the upper limit of the pH of the royal jelly fraction is not particularly limited. However, in view of the resistance of the ultrafiltration membrane, the pH is usually preferably 12.0 or less. In addition, this result shows that by adjusting the pH of raw royal jelly in advance before ultrafiltration, the solubility of hydroxydecenoic acid in the ultrafiltrate can be increased, so when preparing the royal jelly fraction It shows that the recovery rate of organic acids with low solubility such as hydroxydecenoic acid from raw royal jelly can be improved.

<Experiment 4: Allergenicity test of royal jelly fraction>
In Experiment 3, it was shown that the recovery rate of hydroxydecenoic acid and the like was improved by adjusting the pH of the raw royal jelly. Therefore, the allergenicity of the royal jelly fraction prepared by ultrafiltration of the raw royal jelly whose pH was adjusted in advance. A test to investigate the above was conducted as follows. That is, the same raw royal jelly used in Experiment 2 was diluted 5 times with water, adjusted to pH 7.2 with 6N sodium hydroxide, and then prepared by the same preparation method as Test Sample 1 of Experiment 2. A test sample (hereinafter referred to as “pH-adjusted test sample”) (pH-adjusted royal jelly fraction) fractionated using an outer filtration membrane was prepared and added to this pH-adjusted test sample, and the test prepared in Experiment 2 Sample 1 (pH unadjusted royal jelly fraction), Test sample 2 (existing purified royal jelly), and Test sample 3 (raw royal jelly water-diluted solution) were used except that the sample of hydroxydecenoic acid in the amount shown in Table 4 was used. Were examined in the same manner as in Experiment 1. For PCA using rats, the sera of mice immunized with these samples were diluted 10-fold, 20-fold, 100-fold and 200-fold, and the results are shown in Table 4. For the sera of mice immunized with the pH-adjusted test sample or test sample 1, no allergenicity was observed, so only the results of 10-fold dilution and 20-fold dilution were immunized with test sample 2 or test sample 3. Since strong allergenicity was observed for mouse serum, only the results when diluted 100-fold and 200-fold are shown. Table 4 also shows the amount of hydroxydecenoic acid and the mass ratio of hydroxydecenoic acid to protein in each test sample used for immunization.

  As is apparent from Table 4, the allergenicity of royal jelly is determined based on the pH-adjusted test sample (pH-adjusted royal jelly fraction) containing 45 μg of hydroxydecenoic acid or the test sample 1 containing 30 μg of hydroxydecenoic acid. In the mice immunized with (pH unadjusted royal jelly fraction), even when the 10-fold diluted and 20-fold diluted sera were used, none of the mice with sera showing positive PCA reaction were observed. In contrast, when immunized with test sample 2 containing 30 μg of hydroxydecenoic acid (existing purified royal jelly), 100-fold diluted and 200-fold diluted serum containing 45 μg of hydroxydecenoic acid In the case of immunization with test sample 3 (diluted solution of raw royal jelly water), 4 plus each of the serum diluted 100-fold and 200-fold, respectively, was determined as 3 plus (++++) and 1 plus (+). (+++++), 3 plus (+++). Although this result shows that the allergenicity of the existing purified royal jelly (test sample 2) described in the specification of International Publication WO2004 / 21803 is lower than that of raw royal jelly (test sample 3), it is still considerably strong allergenic. Tells you that you have On the other hand, the raw royal jelly ultrafiltrate (pH adjustment test sample) subjected to ultrafiltration after adjusting the pH in advance is allergenic in the same manner as the unadjusted royal jelly fraction (test sample 1). Was found to be reduced.

<Experiment 5: Effect of protein concentration in royal jelly on allergenicity>
From the results of Experiment 2 and Experiment 4, it can be seen that the protein contained in the royal jelly has allergenicity. Therefore, an experiment for examining the effect of the concentration of the protein in the raw royal jelly on the allergenicity was conducted as follows. That is, by adding the internal solution of the ultrafiltration membrane produced during the preparation to the pH adjustment test sample prepared in Experiment 4, the raw royal jelly-derived hydroxydecenoic acid and protein are listed in Table 5. The test sample was prepared so that it might become a mass ratio. The allergenicity of these test samples was evaluated in the same manner as in Experiment 1 by immunizing mice with an amount of each test sample containing 45 μg of hydroxydecenoic acid and evaluated by PCA using rats. It shows together with. In the case of PCA using rats, the serum of mice immunized with a test sample having a hydroxydecenoic acid / protein mass ratio of 1: 0.05 or less protein mass is 100-fold or 200-fold. Since no PCA positive reaction was observed in the diluted serum, only the results of 10-fold dilution and 20-fold dilution are shown. In addition, mouse serum immunized with a test sample having a mass ratio of hydroxydecenoic acid to protein of 1: 0.2 or higher than that is 3 plus for 10-fold and 20-fold diluted sera. Since a strong PCA positive reaction of (+++) or more was observed, only the results obtained when the 100-fold and 200-fold dilutions were used are shown.

  As is clear from Table 5, when immunized with a test sample containing 45 μg of hydroxydecenoic acid, the mass ratio of hydroxydecenoic acid to protein in the test sample was 1: 0.2 or higher than that. Sera from mice immunized with a large proportion of test samples were PCA positive even with 200-fold diluted sera, confirming strong allergenicity in the test samples, and 10 fold in sera from mice immunized with 1: 0.05 test samples. PCA was positive only with diluted serum, and allergenicity was confirmed in the test sample although it was weak. For these test samples, the serum of mice immunized with a test sample with a ratio of hydroxydecenoic acid to protein of 1: 0.02 or less than that of protein, even 10-fold diluted serum PCA was negative and no allergenicity was observed. This result shows that the ratio of hydroxydecenoic acid and protein contained in the raw royal jelly is less than 0.05 parts by weight of protein with respect to 1 part by weight of hydroxydecenoic acid, and preferably the protein with respect to 1 part by weight of hydroxydecenoic acid. This indicates that the allergenicity of royal jelly can be effectively reduced by reducing the content to 0.02 parts by mass or less.

<Experiment 6: Comparison of physiological activities of royal jelly fraction, existing refined royal jelly and fresh royal jelly>
In Experiments 1 to 3, it was confirmed that the royal jelly fraction fractionated with an ultrafiltration membrane retains the collagen production enhancing action of raw royal jelly, so other physiological activities of raw royal jelly The test for confirming whether or not the sample was retained was carried out as follows. That is, regarding the physiological activity of the pH adjustment test sample and the test samples 1 to 3 used in Experiment 4, the presence or absence of the physiological activity shown in Table 6 and its The degree was examined, and the results are also shown in Table 6. In addition, each test used the quantity used as an equivalent amount for all the test samples in terms of raw royal jelly used for the preparation of the test samples. The presence or absence of each physiological activity is the activity (◯) in the case of test sample 3 (raw royal jelly water diluted solution), the activity equivalent to this (◯), the activity but the weaker activity (Δ) and the activity than test sample 3 The evaluation was made in three stages of no (×). In addition, as for the measurement method of the physiological activity shown in Table 6, the measurement of the collagen production enhancing action and the IL-4 production inhibitory action is equivalent to the raw royal jelly used for the preparation of the test sample as the test sample. Was carried out in the same way as in Experiment 1. Moreover, TNF-α used the TNF-α production system by intraperitoneal cells of BALB / c mice stimulated with IFN-γ and lipopolysaccharide described in Experiment 5-2 of International Publication WO2004 / 019971. According to the method, serum IgE inhibitory activity is determined based on the method described in the section of Materials and Methods on pages 175 to 176 of “Natural Medicines”, Vol. 55, pp. 174 to 180 (2001). Instead of using cedar pollen allergen (prepared by Hayashibara Biochemical Laboratories, Inc.). In addition, the anti-atopy action can be achieved by the method described in the section of Materials and Methods (Materials and Methods) in “International Immunopharmacology”, Vol. 3, pages 1313 to 1324 (2003), pages 1314 to 1318. The inhibitory effect on the disease was determined by the method described in the section “Materials and Methods” in pages 94 to 95 of “European Journal of Pharmacology”, volume 410, pages 93 to 100 (2000). Moreover, the presence or absence of the testis function activation action was performed by the method shown below using a hamster according to the method described in Experiment 3 of the international publication WO01 / 060388.

<Method of measuring testicular function activation effect>
A powder feed was prepared by adding the pH-adjusted test sample or test samples 1 to 3 so that the added amount per total mass of the feed was 50 ppm in terms of mass as raw royal jelly. Moreover, it replaced with the test sample which adjusted pH, and prepared the control powder feed which added the same mass anhydrous trehalose (prepared by the method of the below-mentioned Example 5). 50 26-week-old male hamsters were bred for 1 week with a control powder feed, then randomly divided into 5 groups of 10 animals, and each group was given a powder feed or control powder feed with any of the test samples added. The mice were reared for 12 weeks while being freely ingested through a commercially available feeder for powder feed. After the breeding period, the testicular function of each individual was examined by analyzing the testis specimen as follows. That is, first, one testis (right side) was extracted from all individuals and fixed with 15% neutral buffered formalin solution according to a conventional method to prepare testis tissue specimens. A section of this specimen was prepared and stained with hematoxylin-eosin (iocin), and then the seminiferous tube was analyzed histologically under a microscope. Each specimen is visually observed at five magnifications at a magnification of 100 times, and the number of seminiferous tubes in one field of view is measured, and the measured seminiferous tube shows a degenerative change (degeneration / disappearance of sperm cells) , Cavities of microtubules, etc.) and vigorous sculpting with solid tissue structure. For each group, the occupancy rate (%) of active semen tubules relative to all tubules was calculated, and the occupancy rate was compared to that fed with the control powdered feed. (○), the same increase (○), or an increase, but lower (△) or no increase than when the food containing the test sample 3 is ingested (No) ( X) was determined in three stages.

  As is clear from Table 6, the pH-adjusted test sample (pH-adjusted royal jelly fraction) retains the same activity as that of test sample 3 (raw royal jelly water-diluted solution) for any of the physiological activities shown in Table 6. It had been. In addition, test sample 1 (unregulated royal jelly fraction) has a test sample 3 (live test) for the TNF-α production inhibitory action, anti-atopic action and hamster testis function activation action among the seven types of measured physiological activities. The other four types of activities whose activity was lower than that of the diluted solution of royal jelly water) retained the same activity as that of the test sample 3. On the other hand, when test sample 2 (existing refined royal jelly) was used, only a weaker activity than test sample 3 (raw royal jelly water diluted solution) was retained for any of the physiological activities shown in Table 6. . This result shows that the ultrafiltrate obtained by fractionating raw royal jelly with an ultrafiltration membrane (pH adjustment test sample and test sample 1) has a higher physiological activity than that of test sample 2 (existing purified royal jelly). It was found that the ultrafiltrate (pH adjustment test sample) obtained by ultrafiltering raw royal jelly by adjusting the pH in advance was particularly excellent from the standpoint of its retention. .

<Experiment 7: Comparison of whitening effect of royal jelly fraction, existing refined royal jelly and fresh royal jelly>
Carboxylic acids such as hydroxydecenoic acid contained in royal jelly are said to have a tyrosinase activity inhibitory action (see, for example, JP-A-9-315928). Then, the test which compares the whitening effect which a royal jelly fraction and a raw royal jelly have was performed as follows. That is, 4.0 × 10 ⁴ cells / 2 mL / well of B16 melanoma cells suspended in RPMI-1640 medium (sold by Sigma Aldrich) containing 10% FCS (v / v) in a 6-well plate (sold by Becton Dickinson) Cells were seeded to form wells and allowed to adhere to the plate. After removing the culture supernatant, the pH adjustment test samples used in Experiment 4 and test samples 1 to 3 were diluted with RPMI-1640 medium containing 10% FCS (v / v), and hydroxydecene contained in these samples was diluted. It diluted so that the final concentration of an acid might become the density | concentration shown in Table 7, and it added at 4 mL / well. Moreover, as a positive control, kojic acid, L-ascorbic acid 2-glucoside known to suppress melanin production of B16 melanoma cells (trade name “AA2G”, sold by Hayashibara Biochemical Laboratories, Inc., hereinafter “AA” 2G ") and hydroxydecenoic acid (sold by Alfresa Pharma Co., Ltd.) diluted with RPMI-1640 medium containing 10% FCS (v / v) so that the final concentration is as shown in Table 7, Added at 4 mL / well. As a negative control, RPMI-1640 medium containing 10% FCS (v / v) was added and cultured. Incubation was carried out at 37 ° C. under 5% (v / v) CO ₂ for 4 days, and in each well, the medium was replaced with a medium having the same concentration as the sample or the positive control initially added on the second day of the culture. It was. Four days after the start of the culture, trypsin-EDTA (trypsin concentration 0.05%, sold by Gibco Invitrogen) was added at 500 μL / well, and B16 melanoma cells were harvested and collected in a 1.5 mL Eppendorf tube. The tube from which the cells were collected was centrifuged (6000 rpm) for 5 minutes, the supernatant was removed, Dulbecco's physiological saline (PBS (−)) was added, and the cells were washed once. Again, after centrifugation (6000 rpm) for 5 minutes, the supernatant was removed, 1N NaOH was added at 250 μL / tube, the cells were solubilized, and the protein mass was quantified by the Bradford method as in Experiment 2, The total protein amount in each well was determined. Among the NaOH solubilized product, the remainder used for protein quantification was boiled for 30 minutes, and the absorbance at 450 nm was measured (Reference 650 nm). A known concentration of melanin (Sigma) is appropriately diluted, treated with alkali in the same manner, the absorbance is measured, a calibration curve is prepared, and the total in B16 melanoma cells cultured with the test sample, positive control or negative control added The amount of melanin was determined. The total protein amount or total melanin amount when cultured with the addition of the test sample or positive control is divided by the total protein amount or total melanin amount when cultured with the negative control added, respectively, and multiplied by 100, The relative values with the total protein amount or the total melanin amount when the culture is added with the negative control as 100% are determined and are also shown in Table 7.

  As is apparent from Table 7, when cultured in a medium supplemented with a test sample and a positive control, suppression of melanin production depending on the concentration was observed. From the point of strength of suppression of melanin production, the pH adjustment test sample and test sample 1 were the strongest, and kojic acid used as a positive control was also observed to have strong activity. Hydroxydecenoic acid and AA-2G showed only a weak melanin production inhibitory effect at the concentrations used in the test. It was also found that when test sample 2 and test sample 3 were added, the inhibitory effect on melanin production was weaker and the total protein amount was lower than when pH adjusted test sample and test material 1 were added. This result shows that the melanin production inhibitory action of the royal jelly fraction is derived from components other than hydroxydecenoic acid. Furthermore, test samples 1 and 2 show that the test sample 1 exhibits stronger melanin production inhibitory activity than the test sample 2 even when the hydroxydecenoic acid concentration is the same, and conversely, the total protein amount is Because it was high, by removing the polymer fraction by ultrafiltration or the like, the components that inhibited the melanin production inhibitory activity in the raw royal jelly and the components that inhibit cell metabolism were removed or reduced, This indicates that royal jelly has enhanced melanin-inhibiting activity and reduced metabolic inhibition. In addition, when AA-2G and the pH adjustment test sample were added to the medium at the same time, the production of melanin was suppressed more strongly than when each of them was added alone, so the royal jelly fraction of the present invention And AA-2G were judged to be able to suppress melanin production additively.

<Experiment 8: Effect of powdered base on powderization of royal jelly fraction>
The royal jelly fraction contains a large amount of carbohydrates and amino acids, so if the aqueous solution is dried as it is by spray drying etc., browning will be significant and there will be problems with hygroscopicity. Therefore, a test for examining the effect of the powdered base on the powdering of the royal jelly fraction was performed as follows. That is, as shown in Table 8, 37 parts by mass (in terms of solids) of 7 types of dextrin and glucose having different dextrose equivalent (DE) and 3 parts by mass of the royal jelly fraction prepared in Experiment 2 (in terms of solids) And lyophilized by a conventional method to prepare powder preparations of royal jelly fractions. 0.5 g of this powder was dissolved in 5 mL of purified water, and the absorbance at 420 nm was measured. The remaining royal jelly fraction powder was sealed in four aluminum pillow containers for each powder base used for pulverization and stored for 4 weeks under conditions of 60 ° C. and 85% relative humidity. From 1 week to 1 week after the start of storage, one container was opened for each powdered base, dissolved in purified water in the same manner as the start of storage, and the absorbance at 420 nm was measured. The results are also shown in Table 8. Separately, the solubility of each dextrin used as a powdered base and glucose itself in water was confirmed. That is, in a 20 mL beaker (diameter 30 mm), 10 g of either dextrin or glucose shown in Table 8 and 5 g of ion-exchanged water were added, stirred with a stirrer, and visually confirmed whether the dextrin was completely dissolved. . If insoluble matter remains, add ion-exchanged water little by little while continuing to visually check and continue stirring, measure the minimum amount of ion-exchanged water required for the dextrin to completely dissolve, The results are also shown in Table 8. For the test, commercially available dextrin (Matsuya Chemical Co., Ltd., trade name “Paindex # 1”, Sanwa Starch Co., Ltd., trade names “Sandek # 70, # 100, # 150, # 250 and # 300” : Corresponding to samples 1 to 6 in Table 8).

  As is apparent from Table 8, when glucose was used as a powdered base, browning occurred in one week of storage. In addition, when DE28.2 or DE21.3 dextrin was used, the absorbance became 0.3 or more and strong browning occurred after 2 weeks of storage. In contrast, when a dextrin having a DE of 16.7 or less was used, a dextrin or glucose having a DE of 21.3 or more and a DE of 21.3 or more was used as a powdered base even after 4 weeks of storage. Compared to the case, the progress of browning was suppressed. In addition, the solubility of the powdered base itself in water decreased with decreasing DE. This result shows that a dextrin having a DE of 16.7 or less is desirable and a dextrin of 11.5 or less is particularly desirable as a powdering base for pulverizing the royal jelly fraction of the present invention. In addition, considering the solubility of the powder of the royal jelly fraction of the present invention in water, it was determined that a dextrin having a DE of 7.1 or more is preferable to a dextrin having a DE of 6.1.

  Hereinafter, the present invention will be described in more detail with reference to specific examples, but the present invention is not limited to these examples.

<Preparation of royal jelly fraction>
40 kg of deionized water at 4 ° C. was added to 10 kg of fresh royal jelly (produced in Brazil), and the mixture was stirred and dispersed uniformly. Subsequently, this suspension was centrifuged using a continuous centrifuge (model KT-2000G, manufactured by Kubota Corporation) under the conditions of 10,000 × g and a flow rate of 10 L / hr to obtain a supernatant and a precipitate. separated. 6N sodium hydroxide was added to the resulting supernatant to adjust the pH to 7.2, followed by filtration through an ultrafiltration membrane having a molecular weight cut off of 6,000 dalton (Asahi Kasei sales, trade name “AIP2013”). Water was pushed with a small amount of deionized water and separated into an ultrafiltrate and an ultrafiltrate concentrate, and about 40 L of this ultrafiltrate was concentrated to about 5 kg by a vacuum concentration method to obtain a royal jelly fraction. In addition to maintaining useful physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc. possessed by raw royal jelly, this product has a high solubility. In addition, this product retains the useful biological activity of royal jelly, and the allergenicity is reduced compared to that of raw royal jelly and existing refined royal jelly. Therefore, it can be powdered as it is, or can be granulated or tableted and used as a health supplement, and this product can be used for food, drink, feed, feed, pet food, cosmetics. It can be advantageously used for the preparation of compositions such as pharmaceuticals, quasi drugs, and miscellaneous goods compositions.

  When this product was analyzed by the method of Experiment 2, the pH was 7.0, the protein recovery rate was 0.003%, no protein band was detected by SDS-PAGE, and allergenicity (mouse serum immunized with this product was determined). Analysis after 10-fold dilution was not observed. The mass ratio of hydroxydecenoic acid to protein was 1: 0.0001, and the mass ratio of hydroxydecenoic acid to nitrogen was 1: 0.15.

<Preparation of royal jelly fraction>
To 20 kg of raw royal jelly (produced in China, containing 22.8 mg / g of hydroxydecenoic acid and 120 mg / g of protein), 80 kg of deionized water at 4 ° C. was added and stirred to uniformly disperse. Next, 6N sodium hydroxide was added to this suspension to adjust the pH to 7.5, followed by filtration through an ultrafiltration membrane (Asahi Kasei Sales, trade name “AIP2013”) having a molecular weight cut off of 6,000 daltons. Water was pushed with a small amount of deionized water and separated into an ultrafiltrate and an ultrafiltrate concentrate, and about 160 L of this ultrafiltrate was concentrated to about 40 kg at 40 ° C. using a vacuum concentration method. In addition to maintaining useful physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc., the raw royal jelly has a raw royal jelly. The recovery rate of hydroxydecenoic acid contained in this product is 96% with almost no loss, and the solubility of hydroxydecenoic acid is improved compared to raw royal jelly. Therefore, the allergenicity is reduced as compared with fresh royal jelly and existing refined royal jelly, so that it is powdered as it is, or this powder is granulated or beaten. The product can be used as a health supplement, and can be used to prepare compositions such as food, drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi drugs, and miscellaneous goods compositions. Further, the product or the composition containing the product can be used as it is, or added to feeds and feeds such as concentrated feed, roughage, and pet food, or dissolved in drinking water orally. By ingesting males of breeding animals such as stallions, breeding cattle, and breeding pigs, the testicular function of these animals can be activated and the breeding rate can be improved.

  When this product was analyzed by the method of Experiment 2, the pH was 7.0, the protein recovery rate was 0.005%, no protein band was detected by SDS-PAGE, and allergenicity (mouse serum immunized with this product was detected). Analysis after 10-fold dilution was not observed. The mass ratio of hydroxydecenoic acid to protein was 1: 0.0001, and the mass ratio of hydroxydecenoic acid to nitrogen was 1: 0.11.

<Preparation of royal jelly fraction>
To 0.5 kg of fresh royal jelly (produced in China), 2 kg of deionized water at 4 ° C. was added and stirred to uniformly disperse. Subsequently, 6N sodium hydroxide was added to this suspension to adjust the pH to 6.5, and then centrifuged at 10,000 × g to separate into a supernatant and a precipitate. 2.4 L of the obtained supernatant was concentrated to about 0.5 kg by a vacuum concentration method, and this was subjected to gel filtration chromatography using Toyopearl HW-40F gel (available from Tosoh Corporation). Eluted with deionized water, collected a low molecular weight fraction having a molecular weight of 6,000 or less, and concentrated to 0.25 kg at 40 ° C. using a vacuum evaporator (made by Shibata Kagaku Co., Ltd., RE-10E-100 type). Thus, a royal jelly fraction (pH 6.7) was prepared. This product has improved hydroxydecenoic acid solubility compared to raw royal jelly. In addition, this product retains useful physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc. possessed by raw royal jelly, and allergenicity of raw royal jelly or existing purified royal jelly Therefore, it can be used as a health supplement, by pulverizing it as it is, or by granulating or tableting this powder. Moreover, this product can be advantageously used for the preparation of compositions such as food and drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi drugs, and miscellaneous goods compositions. Further, the product or the composition containing the product may be used as it is, or added to feed or feed such as concentrated feed, roughage or pet food, or dissolved in drinking water orally, By ingesting males of breeding animals such as breeding pigs, the testicular function of these animals can be activated and the reproduction rate can be improved.

<Preparation of royal jelly fraction>
4 kg of deionized water at 4 ° C. was added to 0.5 kg of fresh royal jelly (produced in Taiwan), and the mixture was stirred and dispersed uniformly. Subsequently, it centrifuged at 10,000 * g and isolate | separated into supernatant and a precipitate. About 4.4 L of the obtained supernatant was concentrated to about 0.5 kg by a vacuum concentration method, centrifuged at 10,000 × g, and separated into a supernatant and a precipitate. This supernatant was subjected to gel filtration chromatography using a Toyopearl HW-40F gel (sold by Tosoh Corporation). Eluted with deionized water, collected a low molecular weight fraction having a molecular weight of 6,000 or less, and concentrated to 0.5 kg at 40 ° C. using a vacuum evaporator (manufactured by Shibata Kagaku Co., Ltd., RE-10E-100 type). Thus, a royal jelly fraction (pH 6.7) was prepared. This product retains useful physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc. possessed by raw royal jelly, and allergenicity is similar to that of raw royal jelly or existing purified royal jelly. Therefore, it can be used as a health supplement, as it is, or powdered or granulated or tableted. Moreover, this product can be advantageously used for the preparation of compositions such as food and drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi drugs, and miscellaneous goods compositions.

<Manufacture of royal jelly fraction powder>
Commercially available α, α-trehalose hydrous crystals (Hayashibara Shoji Co., Ltd., trade name “Treha”) are used for about 7 hours under conditions of a temperature of 90 ° C. and a pressure of 300 to 350 mmHg using a jacketed rotary vacuum dryer. Dry under reduced pressure. Thereafter, the temperature was returned to normal temperature and the atmospheric pressure was returned to normal pressure, and the product was recovered to obtain anhydrous α, α-trehalose. 8 kg of this anhydrous α, α-trehalose was put into a fluid bed granulator (Fuji Paudal Co., Ltd., model number “20F (G)”), and 9 kg of the royal jelly fraction prepared by the method of Example 2 was sprayed. 9 kg of a dried product of the royal jelly fraction was prepared. Furthermore, this was grind | pulverized with the universal mixer (Dalton Co., Ltd. model number "60MD-rrs"), and the royal jelly fraction containing powder was prepared. This product retains physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc. possessed by raw royal jelly, and allergenicity is compared to raw royal jelly and existing purified royal jelly. Therefore, it can be granulated or tableted as it is and used as a health supplement. Moreover, this product can be advantageously used for the preparation of compositions such as food and drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi drugs, and miscellaneous goods compositions. Further, the product or the composition containing the product may be used as it is, or added to feed or feed such as concentrated feed, roughage or pet food, or dissolved in drinking water orally, By ingesting males of breeding animals such as breeding pigs, the testicular function of these animals can be activated and the reproduction rate can be improved.

<Manufacture of royal jelly fraction powder>
Example 8: 8% of α, α-trehalose hydrous crystals (Hayashibara Shoji Co., Ltd., trade name “Treha”) were put into a fluidized bed granulator (Fuji Paudal Co., Ltd., model number “20F (G)”). 9 kg of the royal jelly fraction prepared by the above method was sprayed to prepare 9 kg of a dried product of the royal jelly fraction. Furthermore, this was grind | pulverized with the universal mixer (Dalton Co., Ltd. model number "60MD-rrs"), and the royal jelly fraction containing powder was prepared. This product retains useful physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc. possessed by raw royal jelly, and allergenicity is similar to that of raw royal jelly or existing purified royal jelly. Therefore, it can be granulated or tableted and used as a health supplement. Moreover, this product can be advantageously used for the preparation of compositions such as food and drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi drugs, and miscellaneous goods compositions. Further, the product or the composition containing the product may be used as it is, or added to feed or feed such as concentrated feed, roughage or pet food, or dissolved in drinking water orally, By ingesting males of breeding animals such as breeding pigs, the testicular function of these animals can be activated and the reproduction rate can be improved.

<Manufacture of royal jelly fraction powder>
In terms of solid matter, with respect to 1 part by mass of the royal jelly fraction prepared by the method of Example 2, a saccharide derivative-containing saccharide of α, α-trehalose (sales by Hayashibara Biochemical Laboratories, Inc., trade name “Tornale ]) 5 parts by mass was added and spray dried by a conventional method to prepare a royal jelly fraction powder. This product retains useful physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc. possessed by raw royal jelly, and allergenicity is similar to that of raw royal jelly or existing purified royal jelly. Therefore, it can be granulated or tableted and used as a health supplement. Moreover, this product can be advantageously used for the preparation of compositions such as food and drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi drugs, and miscellaneous goods compositions. Further, the product or the composition containing the product may be used as it is, or added to feed or feed such as concentrated feed, roughage or pet food, or dissolved in drinking water orally, By ingesting males of breeding animals such as breeding pigs, the testicular function of these animals can be activated and the reproduction rate can be improved.

<Manufacture of royal jelly fraction powder>
To 1 part by mass (converted to solid matter) of the royal jelly fraction prepared in Example 1, 12 parts by mass of a commercially available dextrin (Sanwa Starch Co., Ltd., trade name “Sandeck # 100”) was added and stirred. It melt | dissolved and it lyophilized | freeze-dried by the conventional method, and the royal jelly fraction powder was prepared. This product retains physiological activities such as collagen production enhancing action, IL-4 production inhibitory action, testicular function activation action, etc. possessed by raw royal jelly, and allergenicity is compared to raw royal jelly and existing purified royal jelly. Therefore, it can be granulated or tableted as it is and used as a health supplement. Moreover, this product can be advantageously used for the preparation of compositions such as food and drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi drugs, and miscellaneous goods compositions. Further, the product or the composition containing the product may be used as it is, or added to feed or feed such as concentrated feed, roughage or pet food, or dissolved in drinking water orally, By ingesting males of breeding animals such as breeding pigs, the testicular function of these animals can be activated and the reproduction rate can be improved.

<Health supplement>
2 parts by mass of sucrose fatty acid ester was added to 98 parts by mass of the royal jelly fraction powder prepared in Example 8 and mixed homogeneously, and formed into tablets of about 200 mg per tablet using a tableting machine. This product is a composition that can be easily used and exhibits remarkable effects. This product retains useful physiological activities such as collagen production-enhancing action, IL-4 production-inhibiting action, testicular function-activating action, etc., and whitening action of raw royal jelly. In addition, this product has allergenicity reduced compared to raw royal jelly and existing refined royal jelly, and exhibits a mild sweetness, so it is used daily, maintaining health, enhancing, whitening, skin, beauty, It is useful as a health supplement for anti-wrinkle and anti-aging.

<Health supplement>
According to the method of Example 5, 0.5 part by mass of the royal jelly fraction obtained by the method of Example 2 was added to 8.5 parts by mass of anhydrous α, α-trehalose, and the mixture was stirred and mixed at room temperature. A royal jelly fraction powder was prepared. 9 parts by weight of this powder, 0.5 parts by weight of sugar-transferred hesperidin (Hayashibara Shoji, trade name “Hayashibara Hesperidin S”), 0.5 parts by weight of pullulan (Hayashibara Shoji, trade name food additive “Pullan”) Were mixed to prepare a royal jelly fraction blend composition. This royal jelly fraction blend composition was formed into tablets of about 300 mg per tablet using a tableting machine. This product retains the useful physiological activity of royal jelly and has enhanced whitening effect. In addition, this product has a reduced allergenicity, is a composition that can be easily used and exhibits a remarkable effect. Since this product shows good taste due to its mild sweetness and moderate acidity, it is a health supplement for daily use, such as health maintenance, enhancement, whitening, skin whitening, beauty, anti-wrinkle, anti-aging, etc. Useful as.

<Health supplement>
In accordance with the method of Example 6, a royal jelly fraction powder containing 1 part by mass of the royal jelly fraction obtained by the method of Experiment 1 with respect to 7.5 parts by mass of hydrous crystals α, α-trehalose was prepared. This powder 8.5 mass part, maltitol 1.3 mass part, L-tryptophan 0.2 mass part was homogeneously mixed to prepare a royal jelly fraction blend composition. This product retains the useful physiological activity of royal jelly and has enhanced whitening effect. In addition, this product is a royal jelly fraction composition that has reduced allergenicity, can be easily used, and exhibits remarkable effects. Since this product shows good taste due to its mild sweetness and moderate acidity, it is a health supplement for daily use, such as health maintenance, enhancement, whitening, skin whitening, beauty, anti-wrinkle, anti-aging, etc. Useful as.

<Ice cream>
18 parts by weight of fresh cream (oil content: about 46% by weight), 7 parts by weight of skim milk powder, 51 parts by weight of whole milk, 10 parts by weight of sugar, 4 parts by weight of lactosucrose-containing powder (trade name “milk oligo”), pullulan 2 A mixture of parts by weight and 2 parts by weight of gum arabic was dissolved, sterilized by holding at 70 ° C. for 30 minutes, emulsified and dispersed with a homogenizer, and then rapidly cooled to 3 to 4 ° C. 4 parts by mass of the royal jelly fraction obtained by the above method was added and further mixed, aged overnight, and then frozen in a freezer to obtain an ice cream. This product exhibits moderate sweetness and elegant flavor, retains the useful physiological activity of royal jelly, and enhances the whitening effect. In addition, this product is an ice cream that is effective for maintaining / promoting health, whitening, skin, beauty, anti-wrinkle, anti-aging, etc. because it contains a royal jelly fraction with reduced allergenicity.

<Amazake>
Cook 10 parts by weight of white rice by adding water according to a conventional method, then cool the obtained cooked rice to 55 ° C., mix 30 parts by weight of rice bran prepared by a conventional method and 0.1 part by weight of salt. After maintaining at 50 to 55 ° C. for 8 hours and then cooling to about 25 ° C., 2 parts by weight of the royal jelly fraction obtained by the method of Example 1 was added and mixed, and then packed into a package. And got amazake. This product is a high-quality amazake with good color and flavor, and is useful as a beverage for maintaining and enhancing health, whitening, skin, beauty, anti-wrinkle and anti-aging.

<Health supplement beverage>
500 parts by weight of anhydrous crystalline maltose (Hayashibara Shoji, trade name “Fine-Tose”), 100 parts by weight of the royal jelly fraction obtained by the method of Example 1, 190 parts by weight of powdered egg yolk, 200 parts by weight of skim milk powder, sugar transfer vitamin C (Hayashibara Shoji Co., Ltd., trade name "ASCOFRESH") 2 parts by weight, sodium chloride 4.4 parts by weight, potassium chloride 1.85 parts by weight, magnesium sulfate 4 parts by weight, thiamine 0.01 parts by weight, ascorbic acid A formulation consisting of 0.1 parts by weight of sodium, 0.6 parts by weight of vitamin E acetate and 0.04 parts by weight of nicotinamide was prepared. 25 parts by mass of this formulation was uniformly dispersed and dissolved in 150 parts by mass of purified water, and 150 g each was enclosed in a brown glass bottle. This product retains the useful physiological activity of royal jelly, has reduced allergenicity and supplemented with nutritional sources, so it maintains health, promotes growth, prevents disease, promotes treatment, after sports It can be advantageously used as a health supplement for the purpose of promoting fatigue recovery, whitening, skin beautification, beauty, anti-wrinkle, anti-aging and the like. In addition, this product can be advantageously used as a composition for oral ingestion or tube administration not only for humans but also for animals such as livestock.

<Health supplement>
The following ingredients were kneaded and mixed, extruded and granulated, and fluidized bed dried to prepare basket granules. This granule was packaged by 1.5 grams.
Royal jelly fraction-containing powder prepared in Example 8 30 parts by weight Erythritol 50 parts by weight Ascorbic acid 2-glucoside 7.5 parts by weight Sucrose (Frost Sugar) 6 parts by weight Coenzyme Q ₁₀ 5 parts by weight Vitamin B ₁ 0.1 Parts by weight vitamin B ₂ 0.1 parts by weight vitamin B ₆ 0.1 parts by weight pineapple powder 0.5 parts by weight orange powder 0.2 parts by weight peach flavor 0.5 parts by weight

  This product retains the useful physiological activity of royal jelly, has reduced allergenicity, and supplemented with vitamins, so it maintains health, promotes growth, prevents disease, promotes treatment, after sports It can be advantageously used as a health supplement for the purpose of promoting fatigue recovery, whitening, skin beautification, beauty, anti-wrinkle, anti-aging and the like. This product can be advantageously used not only for humans but also for animals such as livestock.

<Pet food (gummy)>
8 parts by weight of water was added to 30 parts by weight of sugar and dissolved by heating. 50 parts by weight of starch syrup was added thereto and boiled to a Brix sugar content of 85 to 90 °, and then cooled to 80 ° C or lower. Subsequently, 7 parts by weight of gelatin dissolved in 10 parts by weight of water was added to this sugar solution, and further 2 parts by weight of beef extract, 3 parts by weight of 50% by weight citric acid solution, and obtained by the method of Example 1. An appropriate amount of 1 part by weight of royal jelly fraction and perfume was added and mixed uniformly. The resulting blend was dispensed into starch molds and allowed to stand overnight to prepare gummy pet food. This product retains the useful biological activity of royal jelly, and contains a royal jelly fraction with reduced allergenicity and supplemented with nutritional sources, so that pets can maintain their health and grow. It can be advantageously used as a pet food for the purpose of promotion, prevention of diseases, promotion of treatment, etc. In addition, this product can be advantageously used as a composition for oral intake not only for dogs and cats but also for animals such as various pets and livestock. In addition, this product can increase the reproduction rate by activating testicular functions of breeding animals such as stallions, breeding cattle, and breeding pigs by ingesting them orally.

<External skin cream>
Based on the following formulation, a skin external cream was prepared by a conventional method.
(Prescription) (%)
Monostearate polyoxyethylene glycerin 2
Self-emulsifying glyceryl monostearate 5
Eicosanil behenate 1
Liquid paraffin 1.9
Trimethylolpropane trioctanoate 1
1,3-butylene glycol 5
Sodium lactate solution 10
Methyl paraoxybenzoate 0.1
Peach leaf extract 1.5
Purified water 62.2
Royal jelly fraction prepared by the method of Example 1 2

  This product exhibits excellent moisturizing properties, retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, enhances whitening action, and reduces allergenicity Therefore, it is useful as a basic cosmetic for the purpose of whitening, beauty, anti-wrinkle, anti-aging, etc., to keep the skin fresh without concern about causing allergic symptoms.

<Emulsion>
Based on the following formulation, an emulsion was prepared by a conventional method.
(Prescription) (%)
Polyoxyethylene (20E.O.) polyoxypropylene (2E.O.) cetyl alcohol 1
Silicon KF96 (20cs) (Shin-Etsu Chemical Co., Ltd. sales) 2
Liquid paraffin (medium viscosity) 3
1,3-butylene glycol 5
4-n-butylresorcinol 0.3
Glycerin 2
Ethyl alcohol 1
Carboxyvinyl polymer 0.3
Hydroxypropyl cellulose 0.1
2-Aminomethylpropanol 0.1
Glycosylnarindin (manufactured by Hayashibara Biochemical Research Institute,
(Purity of glucosylnardin) 96%) 0.5
L-ascorbic acid-2-glucoside (Hayashibara Biochemical Laboratories, Inc., trade name “AA2G”) 1
Royal jelly fraction prepared by the method of Experiment 2 1
Antioxidant Appropriate amount Preservative Appropriate amount Buffer for pH adjustment Appropriate amount Purified water is added to make the total amount 100%.

  This product exhibits excellent moisturizing properties, retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, enhances whitening action, and reduces allergenicity Therefore, it is useful as a basic cosmetic for the purpose of whitening, beauty, anti-wrinkle, anti-aging, etc., to keep the skin fresh without concern about causing allergic symptoms.

<Lotion>
A lotion was prepared by a conventional method based on the following formulation.
(Prescription) (%)
Sodium hyaluronate (1% aqueous solution) 20
Carbohydrate derivatives of α, α-trehalose 2
Sorbitol solution (60% aqueous solution) 2.1
PH-adjusted royal jelly fraction prepared by the method of Experiment 4 1
Licorice extract 0.5
Chamomile extract 0.05
Sage extract 1
Aloe juice powder 0.001
Citric acid 0.02
Sodium citrate 0.18
Methyl paraoxybenzoate 0.05
Propyl gallate 0.02
Purified water is added to bring the total volume to 100%.

  This product exhibits excellent moisturizing properties, retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, enhances whitening action, and reduces allergenicity Therefore, it is useful as a basic cosmetic for the purpose of whitening, beauty, anti-wrinkle, anti-aging, etc., to keep the skin fresh without concern about causing allergic symptoms.

<Lotion>
A lotion was prepared by a conventional method based on the following formulation.
(Prescription) (%)
Dipotassium glycyrrhizinate 0.2
Citric acid 0.1
Sodium citrate 0.3
Syrup containing a carbohydrate derivative of α, α-trehalose (sales by Hayashibara Biochemicals, Inc., trade name “Tornale”) 2
Ethanol 5
PH adjusted royal jelly fraction prepared by the method of Experiment 4 2
Photosensitive element 201 0.0001
Ethylparaben 0.1
Add purified water to bring the total volume to 100%.

  This product exhibits excellent moisturizing properties, retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, enhances whitening action, and reduces allergenicity Therefore, it is useful as a basic cosmetic for the purpose of whitening, beauty, anti-wrinkle, anti-aging, etc., to keep the skin fresh without concern about causing allergic symptoms.

<Hair restorer>
A hair restorer was prepared by a conventional method based on the following formulation.
(Prescription) (%)
Dipotassium glycyrrhizinate 0.1
Photosensitive Element 301 0.005
Water-containing crystal α, α-trehalose 0.25
Glycerin 2
Carbohydrates containing saccharide derivatives of α, α-trehalose, sold by Hayashibara Biochemical Research Co., Ltd., trade name “Tornale”) 0.01
Royal jelly fraction prepared by the method of Example 2 0.5
Glucose transfer hesperidin (Hayashibara Biochemical Laboratories, Inc.
Product name "Alpha Glucosyl Hesperidin") 0.01
Glycosylnarindin (manufactured by Hayashibara Biochemical Research Institute,
0.1% purity of glucosylnarindin) 0.1
Glucose transfer rutin (Hayashibara Biochemical Research Institute sales,
Product name "Alpha Glucosyl Rutin") 0.01
Indigo extract (sales of Hayashibara Biochemicals, Inc.,
Product name "Ai Luros") 0.5
Seaweed extract 0.02
Assembly extract 3
Button extract 0.1
Ethanol 45
Add purified water to bring the total volume to 100%.

  This product exhibits excellent moisturizing properties and retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, and allergenicity is also reduced. Hair tonic skin that has excellent effects on hair growth, suppresses dandruff and hair loss, and keeps the scalp clean, because it has anti-inflammatory and anti-aging effects on the scalp without causing symptoms, and has strong antibacterial properties. It is useful as an external preparation.

<Shampoo>
A shampoo was prepared by a conventional method based on the following formulation.
(Prescription) (%)
Piroctone Olamine 0.5
Edetate disodium 0.3
Photosensitive Element 201 0.002
Citric acid 0.3
Sodium salicylate 0.2
1,3-butylene glycol 3
Sodium polyoxyethylene lauryl ether sulfate 6.75
Coconut oil fatty acid diethanolamide 2
Lauryl sulfate triethanolamine 10
Polyoxyethylene lanolinic acid (80 EO) 0.5
2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine 10
Hydroxyethylcellulose hydroxypropyltrimethyl ammonium chloride ether 0.8
Royal jelly fraction prepared by the method of Example 2 3
Indigo extract (sold by Hayashibara Biochemical Research Co., Ltd., trade name “Indigo Luros”, 30 1,3-butylene glycol)
% Water extract of green grass) 2
Fragrance 0.2
Add purified water to bring the total volume to 100%.

  This product exhibits excellent moisturizing properties, retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, enhances whitening action, and reduces allergenicity Therefore, it is a shampoo having effects such as whitening, beauty, anti-wrinkle and anti-aging, which keeps the skin fresh without concern about causing allergic symptoms.

<Rinse>
Based on the following compounding prescription, what mixed A component with heat was mixed with what heat-mixed B component, and it emulsified and prepared the rinse by the conventional method.
(Prescription) (%)
<A component>
Liquid paraffin 2.5
Myristic acid 0.5
Cetanol 1.5
Glycerol monostearate 3
Lauroyl glutamic acid polyoxyethylene octyldodecyl ether diester 1
Pyroglutamic acid isostearate polyoxyethylene glyceryl 0.5
<B component>
Syrup containing saccharide derivatives of α, α-trehalose (sales by Hayashibara Biochemicals, Inc., trade name “Tornale”) 3
1,3-butylene glycol 3
Royal jelly fraction prepared by the method of Example 1 3
Photosensitive dye 301 No. 0.01
Lauroyl-L-lysine 2.5
Fatty acid l-arginine ethylpyrrolidone carboxylate 0.5
Stearyltrimethylammonium chloride 0.5
Glucose-transferred rutin (sales by Hayashibara Biochemical Research Co., Ltd., trade name “αG rutin”)
Sodium pyrrolidonecarboxylate 1
Assembly extract 1
Purified water is added so that the total of component A and component B is 100%.

  This product exhibits excellent moisturizing properties, retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, enhances whitening action, and reduces allergenicity Therefore, it is a rinse that has the effects of whitening, beauty, anti-wrinkle, anti-aging, etc., which keeps the scalp fresh without worrying about causing allergic symptoms.

<Toilet soap>
Based on the following formulation, a cosmetic soap was prepared by a conventional method.
(Prescription) (%)
Neat soap obtained by subjecting beef tallow and coconut oil in a mass ratio of 4 to 1 to ordinary saponification and salting-out methods 94.5
Royal jelly fraction powder prepared by the method of Example 8
Maltitol 1
Water-containing crystal α, α-trehalose 0.5
L-ascorbic acid 2-glucoside (Hayashibara Biochemical Laboratories Co., Ltd., trade name “AA2G”) 0.5
Sucrose 0.5
Glucose-transferred rutin (Sold by Hayashibara Biochemical Research Co., Ltd., trade name “αG rutin”) 0.5
Photosensitive element 201 0.0001
Perfume appropriate amount The total amount is 100%.

  This product exhibits excellent moisturizing properties and retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, and allergenicity is also reduced. This soap has the effects of whitening, beauty, anti-wrinkle, anti-aging, etc.

<Bath agent>
Based on the following formulation, a bath preparation was prepared by a conventional method.
(Prescription) (%)
Hydrous crystal α, α-trehalose (Hayashibara Biochemical Laboratories Co., Ltd., for cosmetics) 74.4
Sodium bicarbonate 12.5
Indigo extract (sales from Hayashibara Biochemical Research, Inc., 1,3-
(Not including butylene glycol) 4
Royal jelly fraction prepared by the method of Example 2 3.5
Powder containing α, α-trehalose and bitter juice prepared by the method described in Example 9 of WO 2003/016325 in a mass ratio of 144: 202 5.5
Photosensitive element 201 0.0015
Perfume Appropriate amount Pigment Appropriate amount Make the total amount 100%.

  This product exhibits excellent moisturizing properties and retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibiting action of royal jelly, and allergenicity is also reduced. It is a bath preparation having effects such as whitening, beauty, anti-wrinkle, anti-aging, etc., which keeps the skin fresh without concern of causing symptoms.

<Toothpaste>
Based on the following formulation, a toothpaste was prepared by a conventional method.
(Prescription) (%)
Dicalcium phosphate 30
Hydroxyapatite 10
Calcium carbonate 5
Syrup containing carbohydrate derivative of α, α-trehalose (sales by Hayashibara Biochemicals, Inc., trade name “Tornale”) 28
Royal jelly fraction prepared by the method of Example 1 3
Sodium lauryl sulfate 1.5
Sodium monofluorophosphate 0.7
Polyoxyethylene sorbitan laurate 0.5
Diphenhydramine hydrochloride 0.5
Preservative 0.05
Add purified water to bring the total volume to 100%.

  This product retains useful physiological activities such as collagen production enhancing action and IL-4 production inhibitory action of royal jelly, and allergenicity is also reduced. It is suitable for the prevention and treatment of gum inflammation, taste disorder, swollen gum due to alveolar pyorrhea, inflammation, bleeding, etc., and is a good toothpaste

<Facial tissue>
A treatment agent was prepared by preparing 3 parts by mass of the royal jelly fraction powder prepared by the method of Example 7 and 1 part by mass of glycerin and 6 parts by mass of water. This treatment agent was sprayed uniformly on a facial tissue not subjected to a softening treatment so as to be 1% of the total mass of the tissue paper in terms of solid content and dried. This product retains the useful physiological activity of royal jelly and contains a royal jelly fraction with reduced allergenicity, so that it suppresses skin inflammation and is a sugar of α, α-trehalose. Compared to the non-softening-treated material derivative, the softness and water absorption of the tissue paper are remarkably improved, so it is gentle to the skin and excellent in the feeling of use. In addition, the treatment agent is used to treat paper towels, toilet paper, tissue paper, non-woven fabrics, etc. in the same manner as facial tissue, thereby stimulating the skin when these papers are used. Since it can reduce, and can improve water absorption and a softness | flexibility, it is also optional to utilize in order to improve the usability | use_condition of these papers.

As described above, the present invention is a royal jelly fraction containing 0.05 part by mass or less of a protein derived from royal jelly with respect to 1 part by mass of hydroxydecenoic acid, and has a collagen production enhancing action, A mouse serum obtained by immunizing a mouse with the royal jelly fraction in an amount containing 45 μg of hydroxydecenoic acid, which has useful physiological activity of royal jelly such as IL-4 production inhibitory action and testicular function activation action. The royal jelly fraction, in which allergenicity to the royal jelly fraction is not detected when judged by PCA using rats, has a markedly reduced allergenicity to mammals including humans and is highly soluble in water. Moreover, in addition to retaining the original useful physiological activity of royal jelly, it also has a whitening effect. One in which entirely based on its own findings that is enhanced. Further, since the royal jelly fraction has no fear of causing serious allergic symptoms, mammals including humans can be used simply and comfortably for maintaining and promoting health, whitening and skin beautification. In addition, the royal jelly fraction of the present invention having the above-described features can be mixed with other ingredients to produce various foods, feeds, feeds, pet foods, cosmetics, pharmaceuticals, quasi drugs, miscellaneous goods, etc. Utilization as a composition is also advantageous. The present invention is an invention that exhibits such remarkable effects, and it is a very significant invention that contributes to this field.

Claims (15)

  A royal jelly fraction, which contains a royal jelly-derived protein at a ratio of less than 0.05 parts by mass with respect to 1 part by mass of 10-hydroxy-2-decenoic acid, and has a useful physiological activity possessed by royal jelly. In addition, when the serum of a mouse immunized with the royal jelly fraction containing 45 μg of 10-hydroxy-2-decenoic acid was determined by passive skin anaphylaxis using a rat, the allergenicity to the royal jelly fraction was Royal jelly fraction not detected.   The royal jelly fraction according to claim 1, wherein the biological activity of the royal jelly is a collagen production enhancing action, an IL-4 production inhibitory activity and / or a testis function activating action.   The royal jelly fraction according to claim 1 or 2, wherein 10-hydroxy-2-decenoic acid is contained in an amount of 5 mg / g or more in terms of solid matter.   The royal jelly fraction according to any one of claims 1 to 3, wherein the amount of nitrogen is 0.3 parts by mass or less with respect to 1 part by mass of 10-hydroxy-2-decenoic acid.   The protein band is not detected by naked eye observation by Coomassie Brilliant Blue staining when the royal jelly fraction containing 30 µg of 10-hydroxy-2-decenoic acid is subjected to SDS-PAGE. The royal jelly fraction described.   When the royal jelly fraction is dissolved in deionized water or diluted with deionized water so that 10-hydroxy-2-decenoic acid contained therein is 0.05 mg / mL or more, the pH is 5.0 or more. The royal jelly fraction according to any one of claims 1 to 5.   The royal jelly according to any one of claims 1 to 6, characterized in that after the raw royal jelly is diluted with water, the polymer fraction having a molecular weight of 10,000 daltons or more derived from the raw royal jelly is reduced and concentrated. A method for producing a fraction.   The royal jelly fraction according to claim 7, wherein the pH of a solution obtained by diluting raw royal jelly with water is adjusted to 5.0 or higher before reducing the high molecular fraction having a molecular weight of 10,000 daltons or more derived from raw royal jelly. Production method.   The manufacturing method of the royal jelly fraction powder which adds a saccharide | sugar to the royal jelly fraction manufactured by the method of Claim 7 or 8, and dries and pulverizes.   A composition comprising the royal jelly fraction according to any one of claims 1 to 6 or the royal jelly fraction obtained by the production method according to any one of claims 7 to 9.   The composition according to claim 10, wherein the composition is any of food and drink, feed, feed, pet food, cosmetics, pharmaceuticals, quasi-drugs and miscellaneous goods compositions.   The royal jelly fraction according to any one of claims 1 to 6 or the composition according to claim 10 or 11, wherein the allergenicity is reduced.   The composition according to any one of claims 10 to 12, wherein the composition is solid.   The composition according to claim 13, comprising a non-reducing saccharide together with the royal jelly fraction.   Use of the royal jelly fraction according to any one of claims 1 to 6 as a whitening agent.