JP2015105272A - Purple sweet potato unrefined sake distillation lees extract - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antioxidative material that is more suitable compared with conventional antioxidative materials.SOLUTION: An alcohol extract is obtained by alcohol extraction of distillation lees obtained from unrefined sake of purple sweet potato.

Description

  The present invention relates to an extract obtained from a distilled koji mash of purple koji.

  Purple grape contains a large amount of polyphenols having a plurality of phenolic hydroxy groups in the molecule. Polyphenol is a causative substance that develops a clear purple color, but has recently been reported to have functions such as an antioxidant action and a hormone promoting action to prevent arteriosclerosis and cerebral infarction.

  In particular, Patent Document 1 describes that solid purple koji shochu has an antioxidant effect.

JP2008-092912

  This invention makes it a subject to provide a more suitable antioxidant material compared with the conventional antioxidant material.

  The present inventors have found that an alcoholic extract obtained by subjecting distilled koji obtained by distilling purple koji mash to alcohol has an excellent antioxidant effect, and completed the present invention after further improvements. I came to let you.

That is, the present invention includes, for example, the subject matters described in the following sections.
Item 1.
Alcohol extract obtained by alcohol extraction of distilled koji obtained by distilling purple koji moromi.
Item 2.
Item 2. The alcohol extract according to Item 1, wherein the alcohol is ethanol.
Item 3.
Item 3. A water-soluble solvent extract obtained by subjecting the alcohol extract (preferably an extract concentrate) according to Item 1 or 2 to water-soluble solvent extraction.
Item 4.
Item 3. An extract obtained by extracting the alcohol extract (preferably extract concentrate) according to Item 1 or 2 with acetone or hexane.
Item 5.
Item 5. An antioxidant composition comprising the extract according to any one of Items 1 to 4.
Item 6.
A method for producing a caffeic acid alkyl ester concentrate, comprising extracting an alcohol extract (preferably an extract concentrate) of distilled koji obtained by distilling purple koji moromi with acetone or hexane.
Item 7.
A method for producing a ceramide concentrate, comprising: extracting an alcoholic extract (preferably an extract concentrate) of distilled koji obtained by distilling purple koji mash with acetone or hexane, and collecting the residue.

  According to the present invention, there is provided an alcohol extract obtained by subjecting distilled koji obtained from purple koji mash to alcohol extraction. The alcohol extract has an excellent antioxidant effect. Moreover, the extract obtained by concentrating the alcohol extract and extracting with acetone or hexane is also provided. The said extract has the further outstanding antioxidant effect. In particular, a component exhibiting an antioxidant effect (antioxidant component) is concentrated by the alcohol extraction or the extraction operation with acetone or hexane, which is preferable. In particular, surprisingly, the extract obtained by this method is concentrated with antioxidant components that are hardly contained in purple persimmon itself. Therefore, this invention also includes the manufacturing method of the said antioxidant component concentrate. The antioxidant component concentrate may contain various components, but caffeic acid alkyl esters are particularly preferably concentrated. Almost no caffeic acid alkyl ester is present in purple koji itself, and it is quite surprising that a caffeic acid alkyl ester concentrate can be obtained by the production method of the present invention in this respect.

The TLC analysis result of the neutral lipid class of purple koji shochu acetone extract is shown. The detection reagents are 50% sulfuric acid for A (left) and 1 mM DPPH reagent for B (right). In each spot, 1 represents fatty acid ethyl, 2 represents triglyceride, 3 represents α-tocopherol, 4 represents fatty acid, and 5 represents sterol. The TLC analysis result of the antioxidant component in the acetone extract of purple koji shochu or yellow koji shochu is shown. The detection reagent is DPPH, and the developing solvent is chloroform / methanol / water (65: 16: 2). A is the analysis result of shochu (left lane; jaundice shochu, right lane: purple koji shochu), B is analysis of purple koji (left lane: raw purple koji, right lane: purple koji shochu) The results are shown respectively. The GC-MS analysis result of the TMS derivative of the spot X of FIG. 2 is shown. In the figure, A to B and 1 to 4 indicate the following results, respectively. A ,: Total ion chromatography (TIC), B: TIC enlarged view, 1: B peak 1, 2: B peak 2, 3: B peak 3, 4: B peak 4. The mass spectrum analysis result of a caffeic acid alkylester is shown. The analysis result of the radical scavenging activity of the lipid concentrate of purple glaze or jaundice is shown. The TLC analysis result of a purple potato shochu ceramide concentrate is shown. The developing solvent is chloroform / methanol / water (90: 10: 1). Ceramide NP, ASG, Ceramide AP, SG, and GC are ceramide NP (normal acid-phytosphingosine), acylsteryl glucoside, ceramide AP (α-hydroxy acid-phytosphingosine), steryl glucoside, glucosylceramide, respectively. Show. The result of having analyzed the ceramide content of the ceramide concentrate obtained from purple potato shochu by HPLC is shown. The LC-MS / MS analysis result of the ceramide concentrate obtained from purple potato shochu is shown. The list which classified the molecular species (sphingoid base-fatty acid) of the free ceramide contained in the ceramide concentrate obtained from purple potato shochu based on the result of FIG.

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

  The present invention relates to an alcohol extract obtained by alcohol extraction of a distilled koji obtained from purple koji moromi.

  The purple persimmon used in the present invention is a kind of sweet potato that is purple to the inside (that is, the pulp color is purple). For example, Ayamurasaki, Amurasaki, Murasakimasari, Nakamurasaki, Tanegashima purple, Okinawa-made red koji , Yamakawa Murasaki, Purple Sweet Road, Miyano 36, Kyushu 109, and the like.

  Purple potato moromi is obtained by subjecting purple potato to a saccharification reaction and alcohol fermentation. Alcohol fermentation may be performed after the saccharification reaction, or saccharification reaction and alcohol fermentation may be performed simultaneously. The saccharification reaction is preferably performed using soot. Preferred examples of the koji used include rice koji or koji (black koji, white koji, yellow koji, etc.). Moreover, it is preferable to perform alcoholic fermentation using yeast. Examples of the yeast include wine yeast, shochu yeast, beer yeast, sake yeast and the like, and among them, shochu yeast is preferable. In particular, a known yeast used for producing shochu shochu is preferred. For example, the steamed purple koji can be left at a temperature of 10 to 40 ° C. in the presence of koji and yeast to cause saccharification and alcohol fermentation to produce moromi.

  Distillers are obtained by distilling moromi. Specifically, distillation can be performed after alcohol fermentation to obtain a distillate and a residue obtained as a residue. The distiller may separate into a liquid part and a solid part by performing solid-liquid separation. Separation can be performed by a Kamata filter press, a centrifugal separation method, or the like. In addition, after performing solid-liquid separation before distillation and separating a liquid part and a solid part, each may be distilled. In this case, although the residue obtained from the liquid part and the solid part may be used together as a distillation cake, it is more preferable to use only the residue obtained from the solid part as the distillation cake. In addition, the koji (namely, koji shochu) obtained from the mash which manufactures koji shochu using purple koji can be preferably used as the distillation koji of the present invention.

  The distiller may be subjected to alcohol extraction as it is, but it is preferable to subject the solid part obtained by further solid-liquid separation of the distiller to alcohol extraction. In particular, it is preferable that the solid part is further dried to obtain a dry solid part, which is subjected to alcohol extraction. The dry solid part preferably has a moisture content of 10% or less, more preferably 5% or less. Examples of the drying method include a double drum dryer, a shelf-type ventilation dryer, a slurry dryer, a vacuum drum dryer, a vacuum dryer, a freeze dryer, and the like. From the viewpoint of maintaining the quality of antioxidant components and cost, A drum dryer is particularly preferred. The dry solid part may be pulverized before being subjected to alcohol extraction to be a pulverized dry solid part. The pulverization treatment is preferably performed using a pulverizer capable of preventing a high temperature in a short time, such as a jet mill, a hammer mill, a grinder, or an airflow pulverizer.

  Alcohol extraction can be performed by a known method. For example, a method of immersing a distillation cake (which may be a solid part, a dry solid part, or a powder dry solid part) in alcohol overnight at about 25 to 45 ° C. can be exemplified. As alcohol used for alcohol extraction, a C1-C4 alkyl alcohol is preferable and especially ethanol is preferable.

  The alcohol extract may be the alcohol extract itself or may be a concentrate (alcohol extract concentrate) of the extract. The concentrate may be concentrated to dryness. The concentration method is not particularly limited, and can be concentrated and dried using, for example, an evaporator.

  The present invention also includes a water-soluble solvent extract obtained by extracting the alcohol extract concentrate with a water-soluble solvent. As the water-soluble solvent, for example, water, ethanol, or a mixed solvent of water and ethanol is preferable. Extraction using the water-soluble solvent can be performed by a known method. For example, a water-soluble solvent extract can be obtained by adding a water-soluble solvent to the alcohol extract concentrate and allowing it to stand or stir. The water-soluble solvent extract may be the water-soluble solvent extract itself, or may be a concentrate (water-soluble solvent extract concentrate) of the extract. The concentrate may be concentrated to dryness. The concentration method is not particularly limited, and can be concentrated and dried using, for example, an evaporator.

  In particular, the water-soluble solvent extract contains a large amount of anthocyanin pigments (that is, is concentrated), and the extract can be preferably used as an antioxidant composition. Moreover, it can also be used as an antioxidant composition in combination with the extract and a pharmacologically or food hygienically acceptable carrier. As such a carrier, known ones can be used. In addition, the antioxidant composition containing these extracts is included in the antioxidant composition containing the said extract.

  The present invention also includes an extract obtained by extracting the alcohol extract concentrate with acetone or hexane. An extract obtained by extraction with acetone is more preferable. The extraction with acetone or hexane can be performed by a known method. For example, acetone or hexane is added to the alcohol extract concentrate (the alcohol extract concentrate remaining after the water-soluble solvent extraction operation can be used), and centrifuged (or filtered) one or more times (for example, 2 to 4 times). ), The supernatant (or filtrate) can be recovered and used as an extract. Although not particularly limited, the centrifugation is preferably performed with cooling (for example, 4 ° C.) (particularly, when obtaining a ceramide concentrate as described below, it is preferably performed with cooling). The extract may be the extract (for example, the supernatant) itself, or may be a concentrate (extract concentrate) of the extract. The concentrate may be concentrated to dryness. The concentration method is not particularly limited, and can be concentrated and dried using, for example, an evaporator.

  The extract obtained by extraction with acetone or hexane contains a large amount of fat-soluble polyphenols (for example, fatty acid ethyl ester and caffeic acid alkyl ester) known as antioxidant components (ie, concentrated). ), And the extract can be preferably used as an antioxidant composition. Moreover, it can also be used as an antioxidant composition in combination with the extract and a pharmacologically or food hygienically acceptable carrier. As such a carrier, known ones can be used. In addition, the antioxidant composition containing these extracts is included in the antioxidant composition containing the said extract.

  The present inventors have found for the first time that these extracts (particularly those obtained by extraction with acetone or hexane) contain an alkyl caffeic acid ester as an antioxidant component. Caffeic acid alkyl ester is known as a fat-soluble antioxidant component that hardly exists in nature. It is also known to have an antitumor effect. Accordingly, the present invention provides a method for producing a caffeic acid alkyl ester-containing product or a caffeic acid alkyl ester, which comprises extracting an alcohol extract concentrate of distilled koji obtained by distilling purple koji mash with acetone or hexane, and Moreover, the caffeic acid alkylester containing material obtained in this way is also included. As the caffeic acid alkyl ester, an ester of an alkyl alcohol having 12 to 20 carbon atoms and caffeic acid is preferable. Among the alkyl alcohols having 12 to 20 carbon atoms, those having 14 to 18 carbon atoms are more preferable, and those having 16 to 18 carbon atoms are more preferable. The alkyl of the alkyl alcohol may be linear or branched, but is preferably linear.

  Furthermore, the present inventors have also found that ceramide is concentrated in the residue after extraction with acetone or hexane to obtain an extract as described above. Ceramide is known to have excellent effects such as moisturizing the skin and enhancing the barrier function of the skin. Therefore, this invention also includes the manufacturing method of a ceramide concentrate including extracting the alcohol extraction concentrate of the distilled lees obtained by distilling the mash of purple lees with acetone or hexane, and collect | recovering residues. The ceramide concentrate contains a large amount of free ceramide in addition to glucosylceramide, which is preferable. The ceramide concentrate may be the residue itself, or may be a residue obtained by washing the residue with, for example, water or ethanol (preferably until the color disappears). Further, the residue may be subjected to a drying process.

  The alcohol extract is a raw material for the water-soluble solvent extract, the acetone or hexane extract, and the ceramide concentrate, and contains abundant components contained in the extract or concentrate. . Therefore, the alcohol extract can be used as it is as an antioxidant composition. Moreover, it is useful as a raw material of these extracts or concentrates.

  Furthermore, the antioxidant composition of the present invention includes a composition containing the alcohol extract, the water-soluble solvent extract, the extract with acetone or hexane, or the ceramide concentrate, for example, a pharmaceutical composition. Specific examples include products, cosmetic compositions, food compositions, oral compositions and the like. By applying the composition, an antioxidant effect can be preferably obtained. Moreover, these compositions can be manufactured, for example according to a conventional method.

  Hereinafter, the present invention will be specifically described, but the present invention is not limited to the following examples.

Extraction from purple koji distilled lees From shochu made from koji koji, solid-liquid separated solids were obtained by Kanda filter press. It was dried by a double drum dryer to prepare a dried shochu. 3 L of 99.5 degree ethanol was added to 1 kg, and the mixture was stirred and extracted overnight at 40 ° C. Solid-liquid separation after extraction was performed by suction filtration using a Buchner funnel equipped with a 24 cm SUS funnel and a vacuum pump. Advantech No.1 filter paper was used as the filter paper. The sufficiently filtered ethanol solution was concentrated and dried with a rotary evaporator using an eggplant-shaped flask to obtain an ethanol extract concentrate. Ethanol at the time of concentration was recovered.

  The ethanol extract concentrate weight was 66 g (6.6% of dry shochu). Next, the ethanol extract concentrate (solid portion) was washed with acetone, transferred to a centrifuge tube, and cooled and centrifuged at 4 ° C. (3000 rpm × 10 min). The obtained acetone liquid layer was used as an acetone extract. The acetone extract was concentrated and dried with a rotary evaporator to obtain an acetone extract concentrate. Its weight was 11 g (17% of the ethanol extract concentrate). Ethanol and water were added to the centrifugation precipitate, and the mixture was centrifuged at 15 ° C. The liquid layer and the precipitate portion were separated, and the precipitate portion was centrifuged again by the same operation. Acetone was added to the obtained centrifugal precipitate, and stirring and centrifugation were repeated until there was no coloration in acetone. The resulting precipitate (ceramide concentrate) was collected after drying (1.6% of the ethanol extract concentrate).

  In addition, the same extraction was performed using jaundice (koganesengan) shochu. However, unless otherwise specified, the following analysis was performed on an extract obtained from purple potato shochu.

Analysis of Antioxidant Components in Acetone Extract Fig. 1 shows the results of analyzing the acetone extract concentrate in chloroform / methanol (2: 1) and analyzing it with silicic acid TLC. When the neutral lipid class is detected using hexane / diethyl ether / acetic acid (80: 20: 1) as a developing solvent (A in FIG. 1), a large spot of fatty acid ethyl ester is observed. In addition, fatty acids and sterols are observed. Was detected. When a DPPH solution, which is a radical agent, was sprayed as a detection reagent (FIG. 1B), the presence of a component having a strong radical scavenging activity was observed near the origin.

  The acetone extract concentrate was extracted twice with hexane, the hexane layer was washed with water, concentrated to dryness, redissolved in hexane and subjected to GC-MS analysis, and the fatty acid ethyl ester (FAEE) contained in the extract was analyzed. The fatty acid composition was examined. The results are shown in Table 1.

  Next, polar lipid classes were analyzed using chloroform / methanol / water (65: 16: 2) as a developing solvent (FIG. 2). As detected by DPPH reagent, spot X and spot Y having radical scavenging activity were observed in purple koji shochu, but a trace amount was found in koganesen gun shochu (A in FIG. 2). In addition, this antioxidant component was hardly found in the raw material koji (B in FIG. 2), indicating that it exists specifically in purple koji shochu. From this, it was found that the antioxidant component is produced during shochu when producing shochu using purple koji.

Structural analysis of antioxidant components The acetone extract concentrate was separated by silicic acid TLC, and the spot X was extracted and purified. According to a conventional method, the lipid was subjected to GC-MS analysis as a trimethylsilyl (TMS) ether derivative (FIG. 3). Multiple peaks were detected from spot X. From the results of MS analysis, it was found that peak 1, which is a large component, was caffeic acid hexadecyl ester and peak 4 was caffeic acid octadecyl ester (FIG. 4).

Radical scavenging activity of acetone extract Acetone extract concentrate was dissolved in 2-propanol and added to DPPH ethanol solution for colorimetric determination of radical scavenging activity (Fig. 5). The acetone extract derived from purple koji shochu showed an antioxidant activity 2.5 times that of the koji koji shochu acetone extract at an addition amount of 1 mg to the reaction system. Moreover, the antioxidant activity of 1.5 mg of purple koji shochu acetone extract was equivalent to the activity of 1.0 mg of pure reagent γ-oryzanol.

Analysis of ceramide concentrate When the residue after obtaining an acetone extract from an alcohol extract (the residue is referred to as “ceramide concentrate” hereinafter) by TLC (FIG. 6), in addition to glucosylceramide, A large spot of free ceramide was detected. (Although it is already known that shochu contains glucosylceramide derived from sweet potato and koji mold as raw materials, free ceramide that is extremely rare in nature was concentrated.)
When ceramide concentrate was quantified using HPLC, free ceramide, which was not detected in many other foods and food residues at all, was contained at a high concentration of 275 mg per 100 g of dried koji shochu. It was also shown that more glucosylceramide was contained than other raw materials (Fig. 7).

  Structural analysis of the ceramide concentrate by LC-MS / MS identified 30 ceramide peaks (FIG. 8). When classified into five groups according to the combination of ceramide sphingoid base and fatty acid, ceramide AP to which phytosphingosine and ultralong-chain hydroxy fatty acid were bonded occupied the majority (FIG. 9).

Claims (7)

Alcohol extract obtained by alcohol extraction of distilled koji from purple koji moromi. The alcohol extract according to claim 1, wherein the alcohol is ethanol. A water-soluble solvent extract obtained by extracting the alcohol extract according to claim 1 or 2 with a water-soluble solvent. An extract obtained by extracting the alcohol extract according to claim 1 with acetone or hexane. The antioxidant composition containing the extract in any one of Claims 1-4. A method for producing a caffeic acid alkyl ester concentrate, comprising: extracting an alcoholic extract of distilled koji obtained by distilling purple koji moromi with acetone or hexane. A method for producing a ceramide concentrate, comprising: extracting an alcoholic extract of distilled lees obtained by distilling purple moromi mash with acetone or hexane and recovering the residue.

Images