Extracts and processed products containing isorhamnetin rutinoside

Abstract

To provide a composition that contains cross progenies of Asparagus officinalis and Asparagus kiusianus as an active ingredient.SOLUTION: The present invention relates to extracts or products from the above-ground portions of the cross progenies of Asparagus officinalis and Asparagus kiusianus, wherein the extracts or products include isorhamnetin rutinoside.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an extract and a processed product containing isorhamnetin rutinoside, a composition comprising the extract and / or the processed product, and a method for producing the same.

Background Art

[0002] It is known that a large amount of rutin (quercetin rutinoside) is contained in above-ground parts such as young stems and cladodes of *Asparagus officinalis*, so-called edible asparagus. For example, Patent Document 1 describes that a hot-water extract of stems or leaves of green asparagus contains rutin. Patent Document 2 describes that the above-ground parts of edible asparagus contain rutin, and in particular, cladode tissues contain a large amount thereof. Rutin is a flavonoid having vascular protective and anti-inflammatory effects, and buckwheat tea and the like are marketed claiming to have high rutin content. On the other hand, it is known that isorhamnetin is contained in ginkgo leaves, *Hippophae rhamnoides* and the like, has a wide range of pharmacological effects against cardiovascular diseases and various tumors, has the potential to prevent neurodegenerative diseases such as Alzheimer's disease, and has pharmacological effects against hyperuricemia and pulmonary fibrosis (Non-Patent Document 1).

[0003] Furthermore, Patent Document 3 and Non-Patent Document 2 describe a hybrid (progeny after crossing) of *A. officinalis* and *Asparagus kiusianus*, and Patent Document 3 describes that the hybrid progeny has resistance to stem blight. However, none of these documents describes that the above-ground extract of the hybrid progeny can be a useful raw material for oral compositions such as foods and drinks.

Prior Art Literature

Patent Literature

[0004]

Patent Document 1

[0005] [Non-Patent Document 1] Gong G et. al., Biomed. Pharmacother. (2020), 128:110301. doi:10.1016 / j.biopha.2020.110301. [Non-Patent Document 2] Ito, T., et al., Euphytica (2011), 182:285-294 [Overview of the project] [Problems that the invention aims to solve]

[0006] The offspring of a cross between A. officinalis and A. kiusianus exhibit resistance to stem blight disease, thus reducing the amount of pesticides needed. While their use as a food ingredient was anticipated, their beneficial components were unknown. The present invention aims to provide a composition containing these beneficial components of the offspring of the cross. [Means for solving the problem]

[0007] The inventors of the present invention diligently studied to solve the above problems and found that the aerial parts of the offspring of a cross between A. officinalis and A. kiusianus contain a large amount of isorhamnetin rutinoside, thus completing the present invention. That is, the present invention provides the following extracts, processed products, oral compositions, and methods for producing compositions containing isorhamnetin rutinoside. [1] An extract of the above-ground parts of a hybrid offspring of Asparagus officinalis and Asparagus kiusianus, comprising isorhamnetin rutinoside. [2] The extract according to [1], wherein the above-ground parts include pseudoleaves. [3] The extract described in [1] or [2] above, which is a polar solvent extract. [4] Processed product of the above-ground parts of a hybrid offspring of Asparagus officinalis and Asparagus kiusianus, comprising isorhamnetin rutinoside, and in the form of freeze-dried, heat-treated, and / or pulverized products. [5] The processed product according to [4], wherein the above-ground part includes pseudoleaves. [6] An oral composition comprising the extract described in any one of the items [1] to [3] above and / or the processed product described in item [4] or [5] above. [7] The oral composition described in [6] above, in the form of food or beverage. [8] A method for producing a composition containing isorhamnetin rutinoside, The process of preparing the above-ground parts of the offspring of the cross between Asparagus officinalis and Asparagus kiusianus, The process involves extracting the aerial parts of the aforementioned cross progeny to obtain a composition containing isorhamnetin rutinoside. A manufacturing method that includes this. [9] The manufacturing method according to [8], wherein the above-ground part includes pseudoleaves.

[10] The manufacturing method according to [8] or [9], wherein the extraction step includes a step of extracting the aerial parts of the crossed progeny with a polar solvent.

[11] The manufacturing method according to any one of items [8] to

[10] , wherein the above-ground parts of the crossed progeny are freeze-dried, heated, and / or pulverized. [Effects of the Invention]

[0008] According to the present invention, a composition (extract or processed product) containing isorhamnetin rutinoside can be obtained by using the aerial parts of the offspring of a cross between A. officinalis and A. kiusianus as a raw material. Therefore, the extract or processed product of the aerial parts of the offspring can be used as a raw material for oral compositions such as food and beverages that promote the physiological effects of isorhamnetin rutinoside. [Modes for carrying out the invention]

[0009] The present invention will be described in more detail below. The present invention relates to extracts or processed products of the above-ground parts of the progeny of a cross between A. officinalis and A. kiusianus. A. officinalis is so-called edible asparagus, and various commonly cultivated varieties are known. A. kiusianus is a close relative of A. officinalis and is also called Hamatamabouki. A. officinalis and A. kiusianus can be crossed by any method used in the art, and several hybrids (progeny) obtained by actually crossing them are known. The varieties of the progeny used to prepare the extracts and processed products of the present invention are not particularly limited and may be known progeny, or they may be produced by appropriately crossing A. officinalis and A. kiusianus.

[0010] The above-ground parts of the aforementioned hybrid progeny contain isorhamnetin rutinoside, and as a result of its extraction, the extract of the present invention contains isorhamnetin rutinoside as an active ingredient. Furthermore, since isorhamnetin rutinoside is stably present in the above-ground parts of the aforementioned hybrid progeny, even after processing such as freeze-drying, heating, and / or grinding, the processed product of the above-ground parts of the hybrid progeny still contains isorhamnetin rutinoside as an active ingredient. Isoramnetin rutinoside is a type of flavonoid glycoside and has the following structure in which rutin is methylated. [ka] (Glc: glucose, Rha: rhamnose)

[0011] Since isorhamnetin rutinoside is hydrolyzed in vivo to produce isorhamnetin, it is expected that the action of isorhamnetin can improve cardiovascular diseases, tumors, neurodegenerative diseases such as Alzheimer's disease, hyperuricemia, pulmonary fibrosis, osteoarthritis, hypertrophic cardiomyopathy, and the like. Therefore, the extract and processed product of the present invention can be used for producing products that promote the physiological effects of isorhamnetin rutinoside or isorhamnetin.

[0012] The above-ground part is not particularly limited, and may include, for example, pseudophylls, young stems, lateral branches, or cultivated stems, and preferably contains pseudophylls. The pseudophylls of the progeny obtained by crossing A. officinarum and A. kiusianus contain about 300 to about 700 mg of isorhamnetin rutinoside per 100 g of dry weight, and the content is higher than that of ginkgo biloba leaves and seabuckthorn, so it is preferable as a raw material for providing isorhamnetin.

[0013] Without being bound by any particular theory, for example, while A. officinarum has an efficient synthetic pathway for flavonoids, it is considered that crossing A. officinarum with A. kiusianus allows metabolic enzymes derived from both plants to act cooperatively, resulting in efficient synthesis of isorhamnetin rutinoside.

[0014] The processing means for preparing the processed product of the present invention is not particularly limited as long as a processed product containing isorhamnetin rutinoside can be obtained, and for example, may be freeze-drying, heating, and / or pulverization. That is, the processed product of the present invention may be in the form of a freeze-dried product, a heat-treated product, and / or a pulverized product. The heating conditions are not particularly limited. For example, heating may be performed at about 35° C. to about 130° C. for about 10 seconds to about 30 minutes under humidified conditions (more specifically, steaming may be performed at about 40° C. to about 100° C. for about 15 seconds to about 150 seconds, or autoclaving may be performed at about 115° C. to about 125° C. for about 15 minutes to about 25 minutes), or roasting may be performed at about 100° C. to about 300° C. for about 1 minute to about 10 minutes (or at about 100° C. to about 120° C. for about 1 minute to about 5 minutes). Heating under such conditions can impart a savory flavor to the processed product. Also, the pulverization conditions are not particularly limited, and for example, pulverization into powder may be performed using a pulverizer.

[0015] The extract of the present invention may be prepared by adding an extraction solvent directly to the above-ground part collected from the progeny of the cross, or may be prepared by adding an extraction solvent to the processed product. The extraction solvent for preparing the extract of the present invention is not particularly limited as long as it can extract isorhamnetin rutinoside, and for example, may include polar solvents such as water (such as hot water), alcohols (such as methanol, ethanol, propanol, and isopropanol) or hydrous alcohols thereof, esters (such as ethyl acetate), and ethers (such as dimethyl ether and diethyl ether). That is, in one embodiment, the extract of the present invention may be a polar solvent extract.

[0016] In the above-ground parts collected from the aforementioned cross-pollination progeny, isorhamnetin rutinoside is stably present, and isorhamnetin rutinoside can be extracted in the same way as immediately after collection, even without performing an extraction operation immediately, or after applying loads such as heating, humidification, and hand-kneading. Thus, the above-ground parts of the aforementioned cross-pollination progeny exhibit excellent processing stability with respect to isorhamnetin rutinoside, making them easy to use as a raw material for processed foods and the like. In one embodiment, a decomposition inhibitor of flavonoids such as rutin and isorhamnetin rutinoside may be added to the extraction solvent as needed. The decomposition inhibitor is not particularly limited, but may be, for example, a weak acid such as phosphoric acid and acetic acid.

[0017] The extract of the present invention may further contain other components derived from the aerial parts of the progeny of a cross between A. officinalis and A. kiusianus, such as rutin or kaempferol rutinoside.

[0018] In another embodiment, the present invention also relates to an oral composition comprising the extract and / or the processed product. Since the progeny of a cross between A. officinalis and A. kiusianus have resistance to stem blight (Patent Document 3), it is possible to reduce the amount of pesticides used in the cultivation of such progeny. For this reason, the extract and processed product of the progeny can be advantageously used as raw materials for an oral composition. For example, when the extraction solvent is hot water, the extract can be consumed directly as tea. In a certain embodiment, the oral composition may be in the form of food or beverage.

[0019] The content of isorhamnetin rutinoside in the oral composition of the present invention is not particularly limited, but for example, about 1 × 10⁶ of the total mass of the oral composition. -3 It may be approximately 0.1% by mass, preferably about 5 × 10 -3 ~Approx. 5×10 -2 It is expressed as mass percent.

[0020] The oral composition of the present invention may further contain any excipients or additives commonly used in the art, as long as they do not impair the purpose of the present invention, and may further contain other active ingredients.

[0021] In another aspect, the present invention also relates to a method for producing a composition containing isorhamnetin rutinoside. The production method of the present invention is The process of preparing the aerial parts of the offspring of a cross between A. officinalis and A. kiusianus, The process includes the step of extracting the aerial parts of the aforementioned progeny to obtain a composition containing isorhamnetin rutinoside. The aforementioned progeny and isorhamnetin rutinoside are as described above with respect to the extracts and processed products of the present invention. In some embodiments, the aerial parts of the aforementioned progeny may be freeze-dried, heat-treated, and / or pulverized.

[0022] In one embodiment, the extraction step may include a step of extracting the aerial parts of the crossed progeny with a polar solvent. The polar solvent is as described above for the extract of the present invention. In another embodiment, the composition produced by the manufacturing method of the present invention may be an oral composition such as a food or beverage.

[0023] The manufacturing method of the present invention may further include any steps commonly used in the art, as long as they do not impair the purpose of the present invention, and may further include steps for adding other active ingredients.

[0024] The present invention will be specifically described below with reference to examples, but the scope of the present invention is not limited to these examples. [Examples]

[0025] 1. Measurement of flavonoid content 1 (1) Tested Asparagus Commonly cultivated asparagus varieties (A. officinalis): Welcome to Sanuki no Mezame Hamatamabouki (A. kiusiana): System number T14 Crossed progeny: Strain number WCK (female parent × male parent = Welcome × Hamatamabouki T14) Strain number G4K (Female parent × Male parent = Sanuki no Mezame × Hamatama Bouki T14) Strain number 18K (female parent × male parent = Sanuki no Mezame × Hamatama Bouki T14)

[0026] (2) Experimental method Young shoots of the test asparagus species were harvested from March 18 to 31, 2020 (spring harvest), and again from August 18 to 25 of the same year (summer harvest). In addition, pseudoleaves of the test asparagus species were collected from August 21 to 28, 2020. Each obtained sample was weighed, frozen, and then freeze-dried sequentially. After freeze-drying, the weight was measured, and the samples were powdered using a powerful small-scale pulverizer (Force Mill, manufactured by Osaka Chemical Co., Ltd.) and stored at -80°C until use.

[0027] 200 mg of freeze-dried powder was mixed with 2-4 mL of 70% ethanol (containing 0.1% phosphoric acid) and stirred with a vortex mixer, followed by sonication for 20 minutes. This was then centrifuged (3,500 rpm, 10 minutes) and the supernatant was collected in a separate test tube. This extraction process was repeated for the precipitate, and the supernatants were combined into a single test tube to obtain ethanol extracts of the test asparagus species. Using the same extraction solvent, the extract from young stems was divided to 4 mL and the extract from pseudoleaves to 10 mL, and these were passed through an HPLC pretreatment filter (SEPARA 0.45 μm PTFE, manufactured by GVS Japan Co., Ltd.) to prepare the analytical samples.

[0028] Flavonoids in each analytical sample solution were identified and quantified by high-performance liquid chromatography under the following conditions. Standard materials for rutin (quercetin rutinoside), kaempferol rutinoside, and isorhamnetin rutinoside were obtained from Sigma-Aldrich. Equipment: HPLC system manufactured by JASCO Corporation (pump PU-4180, detector MD-4010) Column: C18-MS-II (4.6mm I.D. × 150mm) Column temperature: 40℃ Eluent A: Water (containing 0.5% phosphoric acid) Eluent B: Acetonitrile (containing 0.5% phosphoric acid) Gradient: 0~10 minutes (B:10%) 10~20 minutes (B:10~20%) 20~38 minutes (B:20~25%) 38~45 minutes (B:25~100%) Flow rate: 0.7mL / min Detection: 350nm (PDA detector) Sample injection volume: 10 or 20 μL

[0029] (3) Experimental results The quantitative results for rutin, kaempferol rutinoside, or isorhamnetin rutinoside in each analytical sample solution are shown in Tables 1 to 3, converted to amounts per 100g of dry weight of the raw material. [Table 1] [Table 2] [Table 3]

[0030] Although isorhamnetin rutinoside was detected in only trace amounts in both common asparagus varieties and Hamatama broom, surprisingly, large amounts of isorhamnetin rutinoside were found in the aerial parts of the offspring of these crosses. In particular, high concentrations of isorhamnetin rutinoside were extracted from the pseudoleaves of these offspring.

[0031] 2. Measurement of flavonoid content 2 Except for using Sanuki no Mezame Violetta or Zenyu Gulliver as the common asparagus variety, line number T70 or T71 as the Hamatama Bouki, and line numbers SR1, SR2, KO, or OK as the progeny of the cross between the common asparagus variety and Hamatama Bouki, the content of various flavonoids was measured in the same manner as in item 1(2) above. The results are shown in Tables 4 to 6. [Table 4] [Table 5] [Table 6]

[0032] Similar to item 1 above, the experiments in this section also showed that isorhamnetin rutinoside is present in small amounts in the above-ground parts of common asparagus varieties and Hamatama broom, but in large amounts in the above-ground parts of their offspring, especially their pseudo-leaf portions.

[0033] 3. Processing stability and extraction efficiency The pseudoleaf portion of the offspring of a cross between A. officinalis and A. kiusianus (female parent × male parent = Sanuki no Mezame × Hamatamabouki T71) is as follows: <1> ~ <7> The powder was prepared by processing it as shown above. The pulverization was performed using a powerful small-scale pulverizer (Force Mill, manufactured by Osaka Chemical Co., Ltd.). <1> 40g of pseudoleaves were freeze-dried on the day of collection to prepare 13.3g of processed material, which was then powdered. <2> 50g of pseudoleaves were steamed for 2 minutes on the day of harvesting, then hand-rubbed. This process was repeated twice, heating on a hot plate set to 110°C for several minutes, then heating again on the hot plate at 110°C for several minutes, and finally heating overnight at 50°C to prepare 15.5g of processed material, which was then powdered. <3> 50g of pseudoleaves were hand-crushed on the day of harvesting, then heated overnight at 30°C under humid conditions of approximately 100% humidity to prepare 15.3g of processed material, which was then powdered. <4> 50g of pseudoleaves were heated at 30°C overnight from the day of collection, freeze-dried to prepare 18.4g of processed material, and then powdered. <5> 40g of pseudoleaves were collected and freeze-dried the following day to prepare 18.3g of processed material, which was then powdered. <6> 50g of pseudoleaves were harvested, steamed for 2 minutes the next day, kneaded by hand, heated on a hot plate set to 110°C for several minutes, kneaded by hand twice, heated again on a hot plate set to 110°C for several minutes, and then heated overnight at 50°C to prepare 20.1g of processed material, which was then powdered. <7> 50g of pseudoleaves were collected and autoclaved at 121°C for 20 minutes the following day to prepare 19.5g of processed material, which was then powdered.

[0034] 70% ethanol (containing 0.1% phosphoric acid) was added to each powder, and 10 mL of ethanol extract was prepared in the same manner as in item 1(2) above. In addition, 10 mL of 90°C hot water was added to each powder, and the mixture was stirred immediately afterward, after 5 minutes, after 10 minutes, and after 15 minutes to prepare hot water extracts. The concentration of isorhamnetin rutinoside in each extract was measured in the same manner as in item 1(2) above. The results are shown in Table 7, converted to the amount per 100 g of dry weight of the raw material. [Table 7]

[0035] Compared to processing methods 1 or 5, processing methods 2-4, 6, and 7 subjected the collected pseudofoliates to stress such as heating and hand-rubbing, but still extracted high concentrations of isorhamnetin rutinoside. Furthermore, as can be seen from the results of processing methods 5-7, isorhamnetin rutinoside was not damaged even if the pseudofoliates were not powdered on the day of collection. In addition, isorhamnetin rutinoside was sufficiently extracted by stirring in hot water without treating the pseudofoliates with 70% ethanol (containing 0.1% phosphoric acid) and ultrasound. <2> The processed material before powdering had a shape similar to that of tea leaves, but even by simply adding hot water to it, isorhamnetin rutinoside could be extracted with an efficiency of about 50% compared to ethanol extract. Therefore, by using the aforementioned pseudoleaves as a raw material, an extract containing isorhamnetin rutinoside at a high concentration can be easily and stably prepared.

[0036] From the above, it was found that a composition containing isorhamnetin rutinoside can be obtained by preparing an extract from the aerial parts of the offspring of a cross between A. officinalis and A. kiusianus. Therefore, the extract from the aerial parts of the offspring can be used as a raw material for oral compositions such as food and beverages that promote the physiological effects of isorhamnetin rutinoside.

Claims

1. A polar solvent extract of the pseudoleaves of a hybrid offspring of Asparagus officinalis and Asparagus kiusianus, comprising isorhamnetin rutinoside.

2. The extract according to claim 1, wherein the polar solvent is hot water, alcohol, or aqueous alcohol.

3. An oral composition comprising the extract described in claim 1 or 2.

4. An oral composition according to claim 3, in the form of a food or beverage.

5. A method for producing a composition containing isorhamnetin rutinoside, The process of preparing pseudoleaves for the offspring of a cross between Asparagus officinalis and Asparagus kiusianus, The process involves extracting the pseudoleaves of the aforementioned cross offspring with a polar solvent to obtain a composition containing isorhamnetin rutinoside. A manufacturing method that includes this.

6. The manufacturing method according to claim 5, wherein the polar solvent is hot water, alcohol, or aqueous alcohol.

7. The manufacturing method according to claim 5 or 6, wherein the pseudoleaves of the crossed progeny are freeze-dried, heat-treated, and / or pulverized.

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