TWI480060B - Scirpusin b-containing composition and method for producing the scirpusin b-containing composition - Google Patents

Description

Composition containing trigonal B and manufacturing method of composition containing trigonal B

The present invention relates to a method for producing a composition containing trigonal B and a composition containing trigonal B derived from Scripusin B derived from passionfruit seeds.

Passion fruit is a plant of Passiflora genus Passiflora, alias: clock fruit, the fruit is sour and sweet, can be used as fruit directly to eat fruit, drink it as pure juice, blended into jelly, cake and other food and beverage, used in Increase the flavor. The seeds of passion fruit are hard and should be removed from the fruit when used as pure juice. Even in the case of fruit and seed diet, the seed coat is directly excreted because it is hard and difficult to digest.

In the past, the seed extract of the passion fruit has been discarded, but the present application has revealed that the seed extract of the passion fruit can be effectively utilized as an active ingredient for inhibiting melanin production and an active ingredient for promoting collagen production (Patent Document 1) 2). Then, it is also disclosed (Patent Document 3) that the active ingredient is a polyphenol compound of picetanucol.

On the other hand, as for the spruceol dimer, Scirpusin B, it has been reported that it has anti-HIV activity (Non-Patent Document 1) and has superoxide anion removal activity (Non-Patent Document 2) Inhibition activity of α-amylase activity (Non-Patent Document 3) and inhibitory activity of xanthine oxidase (Non-Patent Document 4).

[Advanced technical literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open Publication No. 2009-102298

[Patent Document 2] Japanese Patent Laid-Open Publication No. 2009-102299

[Patent Document 3] Japanese Patent Laid-Open Publication No. 2010-30911

[Non-patent literature]

[Non-Patent Document 1] Guo-xun Yang, Jin-tao Zhou, Ya-zun Li, and Chang-qi Hu "Anti-HIV Bioactive stilbene dimers of Caragana rosea." Planta. Med. 71 (2005), p569-571 .

[Non-Patent Document 2] Ting Xiang, Toshio Uno, Fumino Ogino, Cuoqian Ai, Jie Duo, and Ushio Sankawa "Antioxidant constituents of Caragana tibetica." Chem. Pharm. Bull. 53(9) (2005), p1204-1206.

[Non-Patent Document 3] Kyoko Kobayashi, Tamaki Ishihara, Eriko Khono, Toshio Miyase, and Fumihiko Yoshizaki "Constituents of stem bark of Callistemon rigidus showing inhibitory effects on mouse α-amylase activity." Bioi. Pharm. Bull, 29(6) , (2006) p1275-1277.

[Non-Patent Document 4] G. Schmeda-Hirschmann, MI Gutierrez, JILoyola, and J. Zuniga "Biological activity and xanthine oxidase inhibitors from Scirpus californicus (CA Mey.) Steud." Phytotherapy Research, 10, (1996) p683- 685.

As far as the general concept of food safety is concerned, it is preferred that the trigonal B which is formulated into foods such as functional foods and health foods is derived from natural substances. However, it has been known that natural products containing trigonal B are rare in content and only a very low content of the trigonal B composition can be obtained.

Accordingly, an object of the present invention is to provide a composition containing a composition of a natural substance and containing a high content of trigonal B, and a method for producing the same.

As a result of intensive studies in order to achieve the above object, the inventors of the present invention have found that the passion fruit seeds discarded in the conventional useless use are rich in trigonal B and complete the present invention.

In other words, the composition containing trigonal B of the present invention is characterized by being derived from a passion fruit seed and containing trigonal B.

According to the composition containing trigonal P of the present invention, a composition derived from a natural substance containing a high content of trigonal B can be provided. Because it is derived from the composition of natural substances, it is also suitable for cosmetics, pharmaceuticals, and food and beverage products that are formulated into functional foods and health foods.

On the other hand, the method for producing a composition containing trigonal B of the present invention is characterized in that at least one selected from an aqueous alcohol solvent and an aqueous ketone solvent is added to the passion fruit seed, and the trigonal B is extracted. To the aforementioned solvent.

According to the method for producing a composition containing trigonal B according to the present invention, tritercin B is extracted into the solvent by adding at least one selected from the aqueous alcohol solvent and the aqueous ketone solvent to the passion fruit seed. Among them, a composition derived from a natural substance containing a high content of trigonal B is produced in a good yield.

In the method for producing a composition containing trigonal B of the present invention, the seed of the passion fruit is preferably a seed obtained by drying the seed of the passion fruit or the seed of the passion fruit.

Further, in the method for producing a composition containing trigonal B of the present invention, the aqueous alcohol-based solvent is aqueous ethanol or aqueous 1,3-butanediol, and the aqueous ketone solvent is preferably aqueous acetone. According to this, the extraction rate of trigonal B is better.

Further, in the method for producing a composition containing trigonal B of the present invention, after extraction, solvent removal, concentration, spray drying, freeze drying, purification, and the like may be performed.

On the other hand, another aspect of the present invention is to add a food or drink derived from the trigonin B of the passion fruit seed.

Further, another aspect of the present invention is a cosmetic which is added to the trigonin B derived from the passion fruit seed.

According to the composition containing trigonal P of the present invention, a composition derived from a natural substance containing a high content of trigonal B can be provided. Because it is derived from the composition of natural substances, it is also suitable for cosmetics, pharmaceuticals, and food and beverage products that are formulated into functional foods and health foods. On the other hand, according to the method for producing a composition containing trigonal B according to the present invention, tritercin B is added by adding at least one selected from the aqueous alcohol solvent and the aqueous ketone solvent to the passion fruit seed. Since it is extracted into the above-mentioned solvent, it is possible to produce a composition derived from a natural substance and contain a high content of trigonal B.

The composition containing trigonal B of the present invention is obtained from a composition of passion fruit seeds and contains trigonal P. The shape may be liquid, solid, semi-solid, gelatinous or the like. Regarding the content of the trigonal B in the composition containing the trigonal B, for example, a preferred range is 1 × 10 ^{-6 by} mass. From 9% to 99.9% by mass, for example, a more preferable range is from 1 × 10 ^-4 % by mass to 20% by mass, and for example, the optimum range is from 1 × 10 ^-3 % by mass to 5% by mass. Further, the content of the trigonal element B in terms of the solid content is, for example, preferably in the range of 0.01% by mass to 99.9% by mass, and more preferably, the range is 0.05% by mass to 30% by mass, for example, the best. The range is from 0.1% by mass to 20% by mass.

Scirpusin B is represented by the following chemical formula (1)

Passion fruit is a plant of the genus Passiflora, alias: clock fruit, its fruit is sour and sweet, as a fruit directly eat fruit, drink its juice as pure juice, blended into jelly, cakes and other food and beverage, used to increase Flavor. The seeds of passion fruit are hard and should be removed from the fruit when used as pure juice. Even if the fruit is eaten together with the seed, the seed coat is hard and difficult to digest and will be directly excreted. The present invention is based on the discovery that the passion fruit seeds discarded in the past have no use for use are rich in trigonal B. Furthermore, the passion fruit seed used in the present invention can also be used as evidence in the history of the past passion fruit, and even if it is formulated into functional foods such as functional foods and health foods, there is no problem in safety.

The composition containing trigonal B of the present invention is preferably obtained, for example, by directly or pulverizing the raw or dried passion fruit seeds and extracting them with various solvents. In order to increase the extraction rate, the passion fruit seed can be suitably subjected to chemical treatment such as acid or alkali decomposition, enzyme decomposition, and then extracted. Further, according to the method for producing a composition containing trigonal P of the present invention described below, it is possible to obtain a composition containing a high content of trigonal B in a particularly good yield, and therefore the best method by this method is obtained. .

Hereinafter, a method for producing the composition containing triterpene B of the present invention will be described.

In the method for producing a composition containing trigonal B of the present invention, the passion fruit seed can use raw or dried passion fruit seeds. Further, in order to increase the extraction rate, the passion fruit seeds may be suitably subjected to chemical treatment such as pulverization, oil extraction, acid or alkali decomposition, and enzyme decomposition, followed by extraction. The pulverization can be carried out using a known pulverizer. Furthermore, since the passion fruit seeds have a large amount of oil, they may become a paste after being pulverized.

Next, at least one selected from the aqueous alcohol solvent and the aqueous ketone solvent is added to the passion fruit seed, and the trigonal B is extracted into the solvent. At this time, it is preferred to carry out extraction by stirring, vibration or heating under reflux.

As the aqueous alcohol-based solvent, an aqueous solvent such as ethanol, methanol or 1,3-butylene glycol can be used, and particularly aqueous ethanol or aqueous 1,3-butanediol is preferred. Further, as the aqueous ketone solvent, an aqueous solvent such as acetone, methyl ethyl ketone, diethyl ketone or chloroacetone can be used, and particularly aqueous acetone is preferred.

The water content, in the case of aqueous ethanol, is preferably from 20 to 99.9% by volume of ethanol, more preferably from 40 to 99.9% by volume, and most preferably from 60 to 80% by volume. In the case of aqueous 1,3-butanediol, it is preferred that 1,3-butanediol is from 10 to 90% by volume, more preferably from 20 to 60% by volume, and most preferably from 30 to 50% by volume. In the case of aqueous acetone, it is preferably from 20 to 99.9% by volume of acetone, more preferably from 40 to 99.9% by volume, and most preferably from 60 to 80% by volume. Outside the above range, the extraction ratio of the trigonal B has a tendency to deteriorate, which is not preferable. In the following, the description of the water content of the aqueous solvent is simplified, for example, 80% by volume of water containing 20% by volume of ethanol is described as "80% aqueous ethanol". Further, for example, 70% by volume of ethanol containing 30% by volume of water is described as "70% aqueous ethanol". Further, for example, 70% by volume of 1,3-butanediol having a water content of 30% by volume is described as 70% aqueous BG". Further, for example, 70% by volume of acetone containing 30% by volume of water is described as "70% aqueous acetone".

The method for producing a composition containing trigonal B of the present invention can be carried out by extraction using a known vibration device. The temperature condition is about 15 to 40 ° C, and the vibration time is preferably about 0.5 to 3 hours. Also, this extraction can be performed by vibration accompanying ultrasonic vibration. According to this, since the vibration can be effectively supplied to the passion fruit seed which is pulverized and contacted with each of the above solvents, the extraction rate of the extraction of the trigonal B from the passion fruit seed into each of the above solvents can be improved, and the accompanying ultrasonic vibration The vibration can be imparted by a known ultrasonic device or the like.

In the method for producing a composition containing trigonal B of the present invention, the extraction can be carried out by a known method of heating and refluxing using the above aqueous alcohol solvent or aqueous ketone solvent. The heating temperature is about 60 to 95 ° C, and the reflux time is preferably about 1 to 2 hours.

Further, in the method for producing a composition containing trigonal B of the present invention, after the extraction, the solvent may be distilled off under reduced pressure. According to this method, a composition containing no organic solvent can be prepared as a food material, and it can be blended into foods such as functional foods and health foods, and may meet safety standards.

In the method for producing a composition containing trigonal B of the present invention, stepwise extraction may be carried out using a plurality of solvents. Thereby, a composition containing tritercin B containing a high content of trigonal B can be produced more preferably in a yield.

Specifically, for example, after the passion fruit seed is pulverized, any one of the aqueous alcohol solvent and the aqueous ketone solvent is added, and vibration or heating reflux is performed, and trigonal B is extracted into the solvent to obtain the first Extracts. The extract is separated from the residue which cannot be recovered as the extract by centrifugation or the like, and the other one of the above solvents is added to the residue, and vibration or heating reflux is performed, and trigonal B is extracted into the solvent. The second extract was obtained. Then, the first extract and the second extract described above are mixed. Further, it is also needless to say that the second extract may also utilize an extract as a passion fruit seed alone.

In this way, by the stepwise extraction of the plurality of solvents, the passion fruit seed is subjected to the first extraction treatment of the aqueous alcohol solvent or the aqueous ketone solvent, and it is considered that the physical properties and the like are changed to suitable extraction. When the second extraction treatment is continued, the extraction rate can be expected to be improved when the aqueous alcohol-based solvent or the aqueous ketone solvent is used or when a solvent other than the above is used.

The extract of the passion fruit seed obtained by the above method is preferably in the range of 1 × 10 ^-6 % by mass to 99.9% by mass, for example, more preferably, the trigonal P content in the extract is, for example, a better range. 1 × 10 ^-4 % by mass to 20% by mass, for example, the optimum range is 1 × 10 ^-3 % by mass to 5% by mass. Further, the content of the trigonal element B in terms of solid content is, for example, preferably in the range of 0.01% by mass to 99.9% by mass, and more preferably in the range of 0.05% by mass to 30% by mass, for example, preferably. The range is from 0.1% by mass to 20% by mass. This extract may be directly or in a concentrated liquid as a composition containing trigonal B. It is also lyophilized by freeze drying or spray drying, and the powder may be used as a composition containing trigonal B. Moreover, there is no limit to the type of these. The insoluble matter contained in the extract may be suitably removed by filtration or the like. The insoluble matter can be further pulverized into fine particles.

In addition, according to the above-mentioned main extract obtained from the seeds of passion fruit, by ion exchange, size exclusion column chromatography, HPLC method, gel filtration, membrane separation, etc., using trigonal B The indicator is distinguished, purified, etc., and this product can be used as a composition containing trigonal B.

In other words, when the HPLC was performed under the following conditions, the trigonal B obtained from the passion fruit seed was eluted in the vicinity of 12.54 minutes (refer to Fig. 1 and, in Fig. 1, in the vicinity of 8.9 minutes) The dissolved material is picetanusol. Further, when performing mass spectrometry by a mass spectrometer, the signal as an anion mode exhibits m/z 485 (see Fig. 2).

[HPLC conditions]

Pipe column: ODS-3 diameter 4.6mm, length 150mm (made by GL Sciences Co., Ltd.)

Column temperature: 45 ° C

Dissolution conditions: flow rate 0.75 ml / min, 15% acetonitrile (0 min) → 40% acetonitrile (20 min) → 45% acetonitrile (25 min)

UV detection: 280nm

Therefore, in the present invention, the presence of the trigonal B derived from the passion fruit seed can be confirmed by the analysis means exemplified above, and at the same time, it can be appropriately distinguished and purified, and the content of the trigonal element B can be modulated and derived. The passion fruit seed contains the composition of trigonal B.

The trigonal P-derived from the passion fruit seed is added to a pharmaceutically acceptable matrix or carrier as a tablet, granule, powder, solution, powder, granule, capsule, gelatin, etc., except In addition to medicine, it can be used as a cosmetic, such as an ointment, a cream, a gel, a film, a lotion, a cosmetic, and the like.

Further, the trigonal B derived from the passion fruit seed can be ingested into a specific food for construction, a nutritional supplement, a functional food, or the like. Such foods may, for example, be chocolate, biscuits, chewing gum, candy, cookies, candy, candy, and other confectionery desserts; cereals; powdered drinks, refreshing drinks, Beverages, nutritious drinks, carbonated drinks, jelly drinks, etc.; frozen desserts such as ice cream and sherbet. Furthermore, buckwheat noodles, Italian noodles, oolong noodles, plain noodles and the like can also be preferred examples. Further, in the case of a specific food for construction and a nutritional supplement, the powder may be in the form of a powder, a granule, a capsule, a syrup, an ingot, or a sugar-coated tablet.

When the trigonale B derived from the passion fruit seed is used in a food or beverage or a cosmetic, the content of the trigonal B in the food or the cosmetic is preferably, for example, 1 × 10 ^-6 % by mass. To 50% by mass, for example, a more preferable range is 1 × 10 ^-4 % by mass to 20% by mass, and for example, the optimum range is 1 × 10 ^-3 % by mass to 5% by mass. Further, the content of the trigonal element B in terms of the solid content is, for example, preferably in the range of 0.01% by mass to 80% by mass, and more preferably in the range of 0.05% by mass to 30% by mass, for example, preferably. The range is from 0.1% by mass to 20% by mass.

(Example)

Hereinafter, the experiment will be specifically described by way of examples, but the scope of the invention is not limited to the examples.

<Test Example 1> (Recovery/purification/identification of Trigonal B from Passion Fruit)

The seeds of the passion fruit were lyophilized and pulverized (60 to 60 mesh pass), 60 ml of 80% acetone was added, and the mixture was stirred at room temperature to prepare a seed extract. This solution was distinguished by HPLC using an ODS column. The HPLC conditions are as follows.

‧ Pipe column: ODS-3 diameter 20mm, length 250mm (made by GL Sciences Co., Ltd.)

‧ Column temperature: room temperature

‧ Dissolution conditions: flow rate 5ml/min, 10% acetonitrile (0min) → 50% acetonitrile (70min) → 80% acetonitrile (75min) → 80% acetonitrile (90min)

‧ UV detection: 280nm

Figure 3 is a diagram showing the HPLC chromatogram. As a result, it was found that there was a maximum peak near the residence time of 51 minutes. Further, it can be seen that there is a second peak near the residence time of 57 minutes.

What kind of substance is dissolved in the vicinity of the above residence time of 57 minutes is identified in the following manner.

First, the difference in dissolution near the residence time of 57 minutes was obtained, and it was purified by HPLC to a purity of about 97%. For this, HPLC/mass analysis (manufactured by Agilent Technologies, Inc., device name "Agillent 1100 LC/MC System") and NMR analysis were carried out. The results are shown below.

ESI: m/z 485 (M); ¹ H NMR (CD30D, 500 MHz): δ 4.38 (1H, d, J = 6.0 Hz), δ 5.32 ((1H, d, J = 6.0 Hz), δ 6.19 (2H, d, J = 2.2 Hz), δ 6.21 (1H, t, J = 2.0 Hz), δ 6.30 (1H, d, J = 6.0 Hz), 6.58 (1H, d, J = 16.1 Hz) , 6.62 (1H, dd, J = 2.2, 8.5 Hz) 6.66 (1H, d, J = 1.9 Hz), δ 6.68 (1H, d, J = 7.8 Hz), 6.74 (1H, d, J = 2.2 Hz) ) 6.78 (1H, d, J = 8.2 Hz), 6.81 (1H, d, J = 16.4 Hz), δ 6.80 (1H, d, J = 1.9 Hz); ¹³ C NMR: (CD30D, 125 MHz): δ : 58.1, 94.8, 96.8, 102.2, 104.4, 107.3x2, 113.6, 114.0, 116.2x2, 116.3, 118.4, 119.8, 120.0, 123.6, δ: 130.9, 131.0, 134.9, 137.0, 146.3, 146.4, 146.5, 146.6, 147.6 , 159.8x3, 162.7

The above NMR analysis results show that it is a spirulina B represented by the following formula (1).

In addition, the result of the optical rotation measurement of the elution component in the vicinity of 57 minutes (manufactured by JASCO Corporation, device name "P-2200 polarimeter") is [α] _D ²³ : 0.0°, according to Fourier (Fourier) Transform infrared spectrophotometer (manufactured by Japan Spectroscopy Corporation, apparatus name "FT / IR-6200 Fourier-Transform infrared spectrometer ") of infrared absorption spectra, in ^{1605cm -1, 1520cm -1, 1445cm -1} , 1339cm -1, 1282 cm ^-1 , 1197 cm ^-1 , 1156 cm ^-1 , 1116 cm ^-1 , and 1002 cm ^-1 have absorption peaks. This result also has no contradiction with the above identification results. Further, in "Kyoko Kobayashi et al,; Biol. Pharm. Bull. 29 (6), (2006) p1275-1277", analysis results of NMR, optical rotation, infrared absorption spectrum, etc. of trigonal B are also described. .

On the other hand, a component which is eluted in the vicinity of the residence time of 51 minutes has been identified as a picatinol represented by the following chemical formula (2).

From the above, it was found that the above-mentioned seed extract was separated by the peak of the peak trigonal B in the vicinity of 57 minutes in the HPLC analysis, and the seed of the passion fruit did contain a large amount of trigonal B. Furthermore, trigonal B was not detected in the peel and pulp of passion fruit.

<Test Example 2> (Extraction of Trigonal B from Passion Fruit Seed)

Review the method for extracting trigonal B from passion fruit seeds. Specifically, the extraction rate of trigonal B was reviewed by the following extraction method.

(1) Vibration extraction by aqueous ethanol and aqueous acetone

200 ml of 80% aqueous ethanol was added to 60 g of the pulverized seeds, and the mixture was shaken at room temperature for 30 minutes, and extraction was carried out to recover the supernatant. This step is repeated twice. Further, 200 ml of 80% aqueous acetone was added to the residue solid, and the mixture was shaken at room temperature for 30 minutes, and extraction was carried out to recover the supernatant. This step is repeated 3 times. The recovered supernatant was then collected, and the obtained extract was concentrated, made up to 200 ml with 80% aqueous ethanol, and injected into the analytical HPLC with 10 μl thereof. Then, a standard curve was prepared by pre-purification into a high purity (97%) as a standard, and the amount of trigonal B contained in the extract was quantified.

(2) heated reflux extraction by aqueous ethanol

To 300 g of the pulverized seeds, 3 L of 80%, 90% or 95% aqueous ethanol was added, and hot reflux extraction was carried out at 92 ° C for 90 minutes. Then, the obtained extract was concentrated, made up to 200 ml with 80% aqueous ethanol, and 10 μl thereof was injected into HPLC for analysis. Then, the amount of the trigonal B contained in the extract was quantified by a standard curve prepared in advance.

(3) Stirring extraction by aqueous 1,3-butanediol (aqueous BG)

About 10 ml of 20%, 40%, 50%, 60% or 80% aqueous BG was added to 10 g of the hot-dried seed or the pulverized seed, and the mixture was heated and stirred at 80 ° C for 1 hour. Then, a part of the obtained extract was subjected to HPLC analysis, and the amount of trigonal B contained in the extract was quantified by a standard curve prepared in advance.

The results are summarized in Tables 1 and 2 below. Further, in Table 1, (A) the amount of trigonal P content (mg) extracted per 100 g of the seed for extraction, or (B) the content of trigonal B in the solid amount of the extract (% by mass).

As shown in Table 1, the passion fruit seeds were extracted from 220 mg to 363 mg of trigonal B per 100 g. In each extraction method, as the content of the solid content of the extract, the vibration extraction system is 2.5% by mass, the 80% aqueous ethanol is heated and refluxed, and the extraction system is 2.1% by mass, and the 90% aqueous ethanol is heated and refluxed, and the extraction system is 3.2% by mass, 95% aqueous The ethanol heated reflux extraction system was 1.9% by mass. Therefore, in the comparison of the extraction methods, the trifluoroside B is most efficiently extracted by a heated reflux extraction system of 90% aqueous ethanol. In addition, when the content of the trigonal B contained in the seeds of the passion fruit is converted to the highest extraction value, it is known that the passion fruit seed contains trigonal B at a high content of at least about 0.36 mass%. Then, it is also known that it can be extracted efficiently.

Further, in Table 2, (A) the amount of the extracted trigonal element B (mg) per 100 g of the seed for extraction, or (B) in terms of the content of the trigonal B in the extract (μg/mL) Said.

As shown in Table 2, the extract was stirred by aqueous 1,3-butanediol (aqueous BG), and the extract of 91.6 mg to 401 mg of trigonal B was extracted per 100 g of the passion fruit seed. Further, the case of 50% aqueous BG is the result of extraction with intact seeds which have not been pulverized. Although the extraction rate was slightly lowered, it was confirmed that extraction was possible without performing pulverization.

Fig. 1 is a HPLC chromatogram for quantitative analysis of trigonal B derived from passion fruit seeds.

Figure 2 is a mass spectrum of trigonal B derived from passion fruit seeds.

Figure 3 is a graphical representation of the HPLC chromatogram of the extract from the passion fruit seed.

Since the picture in this case is experimental data, it is not a representative figure of this case.

Therefore, there is no designated representative map in this case.

Claims (6)

A method for producing a composition comprising trigonal element B, characterized in that at least one selected from the group consisting of an aqueous alcohol solvent and an aqueous ketone solvent is added as a solvent to the passion fruit seed, and the trigonal element is heated and refluxed. B is extracted into the aforementioned solvent. A method for producing a composition comprising trigonal B, characterized in that aqueous 1,3-butanediol is added as a solvent to passion fruit seeds, and trigonal B is extracted into the solvent under heating. The method for producing a composition containing trigonal B as described in claim 1 or 2, wherein the passion fruit seed is a seed of passion fruit or a seed of passion fruit is dried. seed. The method for producing a composition containing trigonal B as described in claim 1, wherein the solvent is aqueous ethanol or aqueous acetone. A method for producing a composition containing trigonal P as described in claim 1 or 2, wherein the solvent is distilled off after extraction. A use of passion fruit seeds for use as a source of trigonal B.