Preparation of anti-oxidant

[Detailed Description of the Invention]

The new molecular entity shown by following formula [] if the present invention relates to the manufacturing method of the anti-oxidization agent which is added by foodstuffs etc. and prevents the oxidization and also it explains in full detail<img file="JPS628474B2_D0001.tif" />It is related with the method of manufacturing the anti-oxidization agent containing 7beta, 11, 12-trihydroxy 6, 10-(epoxy methano) red-throated loon Eta- 8, 11, and 13-trien 20-ON. Although synthetic additives, such as BHA (burylhydroxyanisole) and BHT (dibutylhydroxytoluene), are conventionally used as an anti-oxidization agent for foodstuffs, in recent years, it is safety, Intermediary Came [ as ] to which use of a natural anti-oxidization agent is requested from fields, such as palatability, research of various natural anti-oxidization agents is done for this reason, the usefulness of natural spices is improved and that research is also done especially widely. The research on search of the antioxidant ingredient which exists in spices separates Carnosol of a bitter taste ingredient from a sage (Salvia carnosaDougl kind) first, and this is molecular formula C.₁₉H₂₆O₄The announcement that it is phenolic ester which Have and contains hydroxy phenanthrene (A. I.White et al.) J. Start in Am.Pharm.Assoc.Sci.Ed., p.31, 33, and 37 (1942), Carnosol is the same substance as bitter taste ingredient Picrossivine from the sage and rosemary (Rosmarinus officinalis L. kind) of a Salvia officinalis L. kind and a Salvia triloba L. kind, The report which showed clearly that it has the structure of following formula () was taken out (C. H.Brieskorn et al., J.Org.Chem., 29, 2293 (1964)).<img file="JPS628474B2_D0002.tif" />It was shown clearly that the Carnotzk acid shown in following formula () existed as a main terpene series ingredient of rosemary, and constant [ of the structure ] was carried out (E. Wenkert et al., J.Org.Chem., 30, 2931 (1965)).<img file="JPS628474B2_D0003.tif" />These days, it washes [ rosemary, the solvent extract of a sage and ], and decolorizes, and high speed liquid chromatography separates an ingredient, It has suggested that an odorless antioxidant substance tasteless besides Carnotzk acid with anti-oxidization nature, Carnosol, and phenyl Dinor exists (S. S.Chang et al., J.Food.Sci., 42, 1102 (1977)). Thus, although many reports of research occur about search of the antioxidant ingredient of spices, it is inapplicable to the foodstuffs which impair the flavor of foodstuffs since spices have a strong characteristic flavor, therefore dislike strong flavors, such as Japanese-style food, to use spices as an anti-oxidization agent as it is. For this reason, extracting an antioxidant component from spices and using this as an anti-oxidization agent is proposed variously, for example, it sets to JP,53-9595,B, By extracting spices (rosemary) with the alkaline solution of direct PH7~10, the classification containing the above-mentioned Carnotzk acid is extracted, and using this as an anti-oxidization agent is proposed. These artificers are attached to a part for the antioxidant drawing obtained from rosemary, and are It was done results about examination variously, They are the extraction processing by an apolar solvent, and washing processing by steam distillation after It was done to rosemary, Extraction processing is performed with the mixed-solution of an apolar solvent and weak alkali solution lower than PH10.5, After making the highly acidic impurities in the above-mentioned residual substance or an extract shift into weak alkali solution, The knowledge of a part for the high drawing of the anti-oxidization effect being obtained was carried out by removing this weak alkali solution, extracting an apolar solvent layer, adding 10.5 or more-PH alkaline water solution subsequently to this apolar solvent layer, and extracting with this 10.5 or more-PH alkaline water solution. That is, what is extracted by ten or more PH(s) according to above-mentioned JP,53-9595,B is although it is an oxidization catalyst substantially and there is a statement that extracting by ten or more PH(s) is not preferred, In the result of these artificers' examination, the amount of [ by 10.5 or more above / PH / alkali ] extraction drawing has the strong anti-oxidization effect, While 7beta, 11, 12-trihydroxy 6, 10-(epoxy methano) red-throated loon Eta- 8, 11, and 13-trien 20-ON which is a new molecular entity contain examination about 1~10% of the weight in this drawing part further about a part for this extraction drawing at It was done and the time, This new molecular entity also has a strong antioxidant action, and the anti-oxidization nature for the extraction drawing by 10.5 or more above [ PH ] alkali carries out the knowledge of saying [ that it is what originates mainly in this new molecular entity ], and is a I got it thing making the present invention. That is, the extract which obtained the residual substance produced by the present invention carrying out steam distillation of the residual substance or rosemary which obtained the extract produced by an apolar solvent extracting rosemary by carrying out steam distillation processing by carrying out extraction processing with the apolar solvent is received, Extraction processing is performed with the mixed-solution of an apolar solvent and weak alkali solution lower than PH10.5, After making the highly acidic impurities in the above-mentioned residual substance or an extract shift into weak alkali solution, This weak alkali solution is removed, an apolar solvent layer is extracted, subsequently to this apolar solvent layer 10.5 or more-PH alkaline water solution is added, extraction processing is performed, and this 10.5 or more-PH alkaline water solution is extracted, The method of manufacturing the anti-oxidization agent containing 7beta, 11, 12-trihydroxy 6, 10-(epoxy methano) red-throated loon Eta- 8, 11, and 13-trien 20-ON is provided. Hereinafter, it explains in detail per present invention. In the manufacturing method of the anti-oxidization agent concerning the present invention, the powder is usually used as rosemary which is materials. The present invention may use this rosemary as materials, extraction processing by an apolar solvent and washing deodorization treatment by steam distillation may be performed first, and extraction processing and washing deodorization treatment may give any first in this case. That is, extraction processing of the rosemary is first carried out with an apolar solvent, and extraction liquid is extracted, The extract obtained is fed into water after solvent distilling off, and it may be made to extract the residual substance of Let's go home for steam distillation. Or Line intermediary extraction liquid is obtained for extraction by an apolar solvent per [ which is obtained by carrying out steam distillation processing of the rosemary first ] steam distillation residual substance, a solvent is distilled off as occasion demands, and it may be made to extract an extract. However, the method of performing steam distillation is first recommended after It was done in extraction processing in respect of operativity, an apparatus scale, etc. As an apolar solvent used for the above-mentioned extraction processing, they are petroleum ether and ligroin, Especially one sort or two sorts or more of n-hexane and ethylene chloride are preferably used for n-hexane, cyclohexane, a carbon tetrachloride, chloroform, dichloromethane, ethylene chloride (dichloroethane, mono-chloroethane, etc.), toluene, benzene, etc. In this case, although the quantity in particular used for extraction is not restricted, it is desirable to consider it as 3~10 weight sections to 1 weight section of materials. As an extraction method, the usual extraction methods, such as the Batches method and the flowing-back method, may be adopted. the intermediary of extraction Line with room temperature is also good -- warming -- although a Line intermediary is also downward good, the method of performing, while making it flow back is efficient. Although extraction time is so preferred that it is long, 2~24 hours may usually be sufficient, and it is preferred to make it repeat the same extraction operation as per extraction residual substance 1~several times. After extraction operation is divided into extraction liquid and an extraction residual substance by the usual methods, such as a fault, centrifugal separation, and Decantation. The extraction liquid obtained by this extraction processing distills a solvent out of this under normal pressure or decompression, extracts an extract, and turns this to a next process. The usual method can be adopted as steam distillation processing, for example, a solid (extract obtained by the extraction processing by materials rosemary or the above-mentioned apolar solvent) is preferably thrown into underwater [ more than 10 time weight ], It agitates, and it heats under after, normal pressure, or decompression distributed underwater, the method of boiling may be adopted, the fragrance ingredient in materials spices vaporizes with vapor by this steam distillation processing, and it is removed from solid dispersion liquid. In this case, vapor can be blown into dispersion liquid at the process of this steam distillation processing, and vaporization of a fragrance ingredient can be promoted. After continuing steam distillation until a fragrance ingredient becomes that there is nothing at steam distillation liquid in Mostly private seal Re (usually 0.5~10 hours), a steam distillation residue is divided into a water layer and solid content (steam distillation residual substance) by the usual methods, such as a fault, centrifugal separation, and Decantation, at the time of heat or cooling, and this solid content is extracted. The residual substance which obtained it by carrying out steam distillation of the extract produced by the present invention extracting a part for the drawing obtained by the processing mentioned above, i.e., rosemary, with an apolar solvent, a part for or the drawing which takes lessons from the extract which obtained the residual substance produced by carrying out steam distillation of the rosemary by carrying out extraction processing with the apolar solvent, and is first obtained by the extraction processing and steam distillation processing by the above-mentioned apolar solvent -- desirable -- this drawing -- dissolving in apolar solvents, such as 1~10-weight section ethyl ether, to 1 weight section per part PH adds alkaline water solution lower than 10.5 to this, and it carries out extraction processing of the part for the above-mentioned drawing with the mixed-solution of a polar solvent and the alkaline water solution in which PH is lower than 10.5. Thus, by processing in weak alkali solution, the highly acidic impurities in the drawing part obtained the extraction processing by an apolar solvent and by carrying out steam distillation shift into weak alkali solution. Saturated sodium bicarbonate solution etc. are mentioned as alkaline water solution in which PH is lower than 10.5. Next, PH to which highly acidic impurities shifted carries out separation removal of the alkaline water solution lower than 10.5, and extracts the above-mentioned apolar solvent layer. 10.5 or more-PH alkaline water solution is added to this apolar solvent layer, and extraction processing is performed. The antioxidant component containing the new molecular entity shown by the above-mentioned [] formula by this shifts to the alkaline water solution side, therefore the alkaline water solution layer after extraction processing is separated from an apolar solvent layer, and a part for the high drawing of the anti-oxidization effect (present invention anti-oxidization agent) is certainly obtained by extracting an alkaline water solution layer. It is preferred to use alkaline water solution at a rate of 0.1~2 copies to one copy of above-mentioned apolar solvent layer in this case. In order to avoid contact with oxygen in the air at the time of extraction, it is desirable to operate it in nitrogen or an inactive gas air current. Although what dissolved sodium hydroxide, potassium hydrate metal hydroxide, or alkaline-earth-metals hydroxide in water can use it as alkaline water solution used for this extraction processing, Anyway, it is required to use 10.5 or more PH(s), preferably the thing adjusted to the range of 10.5~11.0, and the antioxidant ingredient containing above-mentioned new molecular entity [] is certainly extracted by carrying out extraction processing using the alkaline water solution of this PH range. On the other hand, even if PH carries out extraction processing with alkaline water solution lower than 10.5, the antioxidant ingredient containing new molecular entity [] is not extracted satisfactorily, and the object of the present invention is not achieved. Although a part for the antioxidant drawing used as an anti-oxidization agent in this extraction processing by extracting after It was done 10.5 or more-PH alkaline water solution (the antioxidant ingredient containing new molecular entity [] shifts and contains in this) is obtained, In this case, after making this alkaline water solution into neutralization or acidity from the thing neutralized from acid, or acid, it is preferred to use the residual substance which extracts an antioxidant ingredient with an apolar solvent and is produced from that extraction liquid by distilling off a solvent. In thus, the inside of the antioxidant drawing part (present invention anti-oxidization agent) obtained, The extraction and steam distillation processing by an apolar solvent are performed to rosemary, respectively, Next, extraction processing is performed with the mixed-solution of an apolar solvent and weak alkali solution lower than PH10.5, After making the highly acidic impurities in the above-mentioned residual substance or an extract shift into weak alkali solution, This weak alkali solution was removed, the apolar solvent layer was extracted, subsequently to this apolar solvent layer 10.5 or more-PH alkaline water solution was added, and extraction processing by 10.5 or more-PH alkaline water solution was performed, New molecular entity 7beta, 11, 12-trihydroxy 6, 10-(epoxy methano) red-throated loon Eta- 8, 11, and 13-trien 20-ON shown by the antioxidant ingredient in rosemary, especially the above-mentioned [] formula shift certainly, This new molecular entity contains about 1~10% of the weight, and it has the strong anti-oxidization power which originates mainly in this new molecular entity. It is in order to obtain the new molecular entity of [] type from a part for this antioxidant drawing, A part for this antioxidant drawing (in this case, after making alkaline water solution into neutrality or acidity) the extract produced by an apolar solvent extracting -- using it -- it applies to silica gel restoration column chromatography, Fractionation in the thin classification of 110 classification by using the mixed solvent of 90:10 (the same [ a capacity ratio and the following ]) of benzene and acetone as deployment liquid, and takes out the 60th classification from the 16th classification. The cotton intermediary can get Carnosol publicly known as an antioxidant ingredient from these 16th classification in the 38th classification, and it can approach with this and can obtain declared new molecular entity []. That is, Carnosol is obtained by re-crystallizing with benzene the solid of the 16~28th classification obtained by removing deployment liquid, The solid of the residual substance which condensed and obtained the benzene liquid except Carnosol, and the 29~38th classification is mixed, This is applied to silica gel restoration column chromatography, and it develops with the mixed solvent of 90:10 of benzene and acetone, and Fractionation in 15 classification, Carnosol is obtained, deployment liquid is removed from the 10~11th classification, and declared new molecular entity [] is obtained from the 7~8th classification by re-crystallizing with acetone. It is while new molecular entity [] is obtained by re-crystallizing the solid of the 39~60th classification in the first chromatography with benzene, and re-crystallizing this crystal with acetone, The solid which removed and obtained benzene from this benzene fusible part is covered over silica gel restoration column chromatography, It develops in six classification with the mixed solvent of 90:10 of benzene and acetone, that 4th classification is taken out, deployment liquid is removed, and this new molecular entity [] is obtained also by re-crystallizing with acetone. The characteristic of this new molecular entity and the anti-oxidization effect are as the reference example mentioned below, and have the anti-oxidization power of being twice [ about ] many as publicly known Carnosol. The anti-oxidization agents of the present invention containing new molecular entity [] are fish oil, lard, and Tharaud, They are oil, cottonseed cake oil, and Sir flower oil to A head, chicken oil, soybean oil, and Sweetness, Rice bran oil, corn oil, coconut oil, palm oil, sesame oil, cacao oil, castor oil, Oil and fats, the butter, cheese, margarine which use Motion, such as peanut oil, and a plant as materials, A shortening, mayonnaise, a dressing, a ham, a sausage, potato chips, a fried rice cracker -- it lifts, and it may be used for them by cosmetics, medical supplies, etc., such as ramen noodles, curry roux, soy sauce, soft drinks, alcohol, fruit wine, catsup, Jam, fish meat or Accumulated meat boiled fish paste, other foodstuffs or hair cosmetics, a skin cosmetic, and inner-mouth cosmetics, being added. In this case, the whole thing of especially the amount of addition of a present invention compound considered as 0.001~0.1 % of the weight 0.0005~1% of the weight is preferred as a dry article. The anti-oxidization agent concerning the present invention is starch by necessity, although this may be used as it is, Diluent bases, such as gelatin, etc. may be added, and it may be made the form of the shape of powder, and the shape of granulation, and dissolution distribution may be carried out at oil and fats, ethanol and propylene glycol, glycerin, or these mixtures, and it may be made a liquefied form. The anti-oxidization agent of the present invention may contain Synelist ingredients, such as citrate, etc. if needed. Even if as for an anti-oxidization agent by the present invention fragrance of rosemary does not have Mostly, either, in spite of anti-oxidization power's being high and acquiring it from rosemary as mentioned above, and it blends with foodstuffs etc., Mostly does not impair the flavor. and -- since it is obtained from natural rosemary by dissociating -- the safety of a synthetic additive agent -- a problem -- it can be conveniently used to foodstuffs including oil and fat products, cosmetics, medical supplies, etc. now [ intermediary To have ]. Next, an example and a comparative example are shown and the present invention is explained still more concretely. An example, a comparative example 500 g of dried leaves of rosemary are ground, n-hexane 1.2 is added, and it is It was done about churning extraction at room temperature overnight. After the suction fault separated liquid, the n-hexane of the amount of these was added to the residual substance, and it processed similarly. This operation was performed 3 times, 3 times of extraction liquid was collected, the solvent was distilled off and condensed under decompression, and n-hexane extract 19.71g was obtained. Next, after feeding this extract 19.71g into 200 ml of water and removing 2.75 g of It was done volatility classification for steam distillation, the steam distillation residue was passed and it divided into liquid and a residual substance. This residual substance was extracted, it dissolved using 400 ml of ethyl ether, subsequently 150 ml of 2N-aqueous hydrochloric acid extracted the ether layer twice, and it separated the chloride layer from the ether layer. After washing this ether layer twice, it extracted 3 times with 150 ml of saturated sodium bicarbonate solution in the nitrogen air current. The ether layer was washed twice after separating a saturated sodium bicarbonate aqueous solution layer, and, subsequently it extracted 4 times with 150 ml of 1N-sodium hydroxide solution in the nitrogen air current in a similar manner, and separated the sodium hydroxide aqueous solution layer from the ether layer. This ether layer was washed twice, with anhydrous magnesium sulfate, after dryness, was condensed and obtained a part for neutral drawing 10.2g. On the other hand, after making this into alkalinity in 4N-sodium hydroxide solution, the above-mentioned chloride layer performed extraction processing with ethyl ether, carried out separation extraction of the ether layer, distilled off the solvent, and obtained a part for basic drawing, but the quantity is a trace grade, and it is Oh. The above-mentioned saturated sodium bicarbonate aqueous solution layer was acidified with 4N-chloride solution, and ethyl ether extracted it, it condensed the ether layer after a flush and dryness, and obtained a part for highly acidic drawing 0.049g. It acidified with 4N-chloride solution, and ethyl ether extracted, it washed and dried, the ether layer was condensed, and the above-mentioned sodium hydroxide aqueous solution layer also obtained a part for weak acidic drawing 1.9g (present invention anti-oxidization agent). New molecular entity [] contained the result of having analyzed a part for this weak acidic drawing, about 2.5%. For comparison, it was operated like the above from the steam distillation volatilization part of the n-hexane extract, and a part for a part for neutral drawing and weak acidic drawing was obtained, respectively. Next, each ingredient obtained with the described method was respectively added 0.01% of the weight to lard, and the peroxide number (POV) was measured by the AOM method (oil recovery study, the 19th volume, No. 5, P62). A result is shown in the 1st table and Drawing 1.

[Table]

[Table]

In Drawing 1, reference mark A, B, and C, D, E, and F are as being shown in the 1st table, respectively, and A' is a peroxide number at the time of adding with lard a part for the weak acidic drawing obtained from the n-hexane extract (present invention anti-oxidization agent) 0.005% of the weight. clear from the result of the 1st table and Drawing 1 -- as -- a part for weak acidic drawing, i.e., a present invention method, -- therefore, the knowledge of only the amount of [ which was obtained ] drawing having high anti-oxidization power was carried out. Next, a reference example explains the manufacturing process of new molecular entity 7beta, 11, 12-trihydroxy 6, 10-(epoxy methano) red-throated loon Eta- 8, 11, and 13-trien 20-ON, and its anti-oxidization effect. Reference example A part for weak acidic drawing 1.42g obtained in the example is taken, and it is silica gel restoration column chromatography (3.0 cm phix24 cm) about this. It applied to silica gel 80g and 0.5 ml/min, and using the deployment liquid which is 90:10, the ratio of benzene to acetone divided into 110 classification (100 ml of one classification), took out the 16~60th classification before long, removed deployment liquid, and obtained each solid. 348 mg of solids of the 16~28th classification were melted with 3 ml of benzene, from this, it was made to re-crystallize and 180 mg (this checked that it was Carnosol by instrumental analysis.) of crystals of the shape of a colorless needle of 233 degrees of melting points C were obtained. The residual substance which condensed and obtained the benzene liquid except the upper Carnosol, and 108 mg of solids of the 29~38th classification are mixed, It is silica gel restoration column chromatography (2.0 cm phix17 cm) about 274 mg of this solid. It applies to silica gel 20g and 0.5 ml/min, and performs time [ 2nd ] Fractionation operation, It develops with the mixed-solution of 90:10 of benzene and acetone, and divides into the classification (10 ml of one classification) of 15, The thing excluding the solvent from the 7~8th classification was re-crystallized with benzene, and 120 mg of Carnosol were obtained, and except for the solvent, it re-crystallized with acetone and obtained 26 mg of declared new molecular entities from the 10~11th classification. Melted 107 mg of solids obtained from the 39~60th classification of the first Fractionation with 1 ml of benzene, and it was made to re-crystallize from this, and divided into the crystal and the benzene meltable part, and the crystal was re-crystallized with acetone and obtained 10 mg of the target new molecular entities. On the other hand, it is silica gel restoration column chromatography (1.5 cm phix11 cm) about 80 mg of solids obtained except for the benzene of benzene solution. It applied to silica gel 8g and 0.35 ml/min, the 3rd Fractionation was performed with the mixed-solution of 90:10 of benzene and acetone, it divided into the classification (7 ml of one classification) of 6, the solid obtained from the 4th classification was re-crystallized with acetone, and 10 mg of the target new molecular entities were obtained. This new molecular entity 7beta, 11, 12-trihydroxy 6, 10-(epoxy methano) red-throated loon Eta- 8, 11, and 13-trien 20-ON show following character and instrumental analysis value.

[Table]

[Table]

Next, BHABHT which is antioxidant substance alpha-tocopherol and the synthetic antioxidant which were obtained from the new molecular entity obtained by the above-mentioned method, and Carnosol and a product of nature is used, and it is I got it about the controlled study of the effect of anti-oxidization nature by the AOM method (oil recovery study, the 19th volume, No. 5, p62). The lard which does not contain an antioxidant substance as blank was taken, the sample was added to this, respectively, and the peroxide number (POV) was measured. The result is shown in the 2nd table and Drawing 5.

[Table]

[Brief Description of the Drawings]

The graph which shows the anti-oxidization power for other drawing of the anti-oxidization agent and rosemary from which Drawing 1 was obtained by the present invention method, It is a graph which shows the anti-oxidization power of the new molecular entity contained in the anti-oxidization agent from which obtain infrared resonance Spector of the new molecular entity which contains Drawing 2 in a present invention oxidizer, and Drawing 3 with the nuclear magnetic resonance spectrum of the compound, Drawing 4 is obtained with the mass spectrum of the compound, and Drawing 5 is obtained by a present invention method, and a publicly known anti-oxidization agent.