Hibiscus×titanbicus brewed vinegar and method for producing the same, and beverage

Abstract

To provide a new processed food prepared from Hibiscus×Titanbicus capable of exhibiting physiological effects, thereby promoting effective use of resources.SOLUTION: The Hibiscus×Titanbicus brewed vinegar is a brewed vinegar obtained by acetic acid fermentation of flowers (including buds) of Hibiscus×Titanbicus, having a quercetin glycoside concentration of 10 mg / 100 mL or more and an acidity of 4.0% or more in terms of acetic acid, and containing isoquercitrin, rutin, and hyperoside, wherein the concentration of isoquercitrin is the highest among them.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] The present invention relates to processed foods made from titan bicus as a raw material, and more particularly to a brewed vinegar made from titan bicus, a method for producing the same, and a beverage containing titan bicus brewed vinegar. [Background technology]

[0002] Traditionally, brewed vinegar, which is produced by acetic acid fermentation of raw materials such as grains, fruits, and vegetables, has been known as vinegar. The main types of brewed vinegar are grain vinegars, such as rice vinegar, rice black vinegar, and barley black vinegar, as well as fruit vinegars, such as apple cider vinegar and grape vinegar. In recent years, various fruit vinegars have been commercially available as health drinks due to their pleasant taste. The acetic acid contained in vinegar is known to have effects such as fatigue recovery, blood sugar reduction, and metabolic improvement, as well as an antihypertensive effect. For example, there are reports that an acetic acid solution prepared to have the same acidity as barley black vinegar also suppressed blood pressure in rats, suggesting that acetic acid may be one of the factors contributing to the antihypertensive effect of brewed vinegar.

[0003] Among the many varieties of horticultural plants, Titanobicus is known for its gigantic flowers. Titanobicus grows vigorously, is easy to cultivate, and is popular for its overwhelming presence. Titanobicus flowers can also be used for food, and in recent years they have also been produced as edible flowers. In the production of edible flowers, harvested buds just before blooming, or flowers artificially bloomed from these buds by temperature treatment, are shipped to customers and then consumed.

[0004] However, for example, buds that are not of the standard size, buds that have been damaged during cultivation, or flowers that have bloomed due to delayed harvesting cannot be shipped for consumption as raw food, and so have traditionally been discarded. Therefore, using these discarded flowers and buds of Titanobicus as ingredients for processed foods is preferable from the perspective of effective resource utilization.

[0005] In recent years, various functions of natural plant extracts have been reported (see, for example, Patent Document 1), but to date, there have been few reports on the components and physiological activities contained in titan bicus, and it has not been used as a processed food. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Publication No. 2018-150262 Summary of the Invention [Problem to be solved by the invention]

[0007] The present invention aims to provide a new processed food using titan bicus, which is expected to exhibit physiological effects, and to make effective use of resources. [Means for solving the problem]

[0008] The titanbicus brewed vinegar of the present invention is a brewed vinegar obtained by acetic acid fermentation of titanbicus flowers (including buds), and is characterized by having a quercetin glycoside concentration of 10 mg / 100 ml or more and an acidity of 4.0% or more in terms of acetic acid.

[0009] In the present invention, "Titanbicus flowers (including buds)" is a concept that encompasses not only flowers in an open state but also unopened flowers (buds), and does not necessarily mean that buds are actually contained. Furthermore, the form of "Titanbicus flowers (including buds)" is not particularly limited, and includes not only the form of flowers or buds, but also processed products (e.g., powders) and extracts thereof; the same applies hereinafter.

[0010] In one embodiment of the present invention, the quercetin glycosides are selected from the group consisting of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb. Q3MG is quercetin with 6-malonylglucose bound to the 3-position, and Q3Samb is quercetin with sambubiose, a disaccharide, bound to the 3-position (see Figure 1).

[0011] The brewed vinegar contains isoquercitrin, rutin, and hyperoside, and is characterized by having the highest concentration of isoquercitrin among these.

[0012] The method for producing brewed vinegar from titanbicus of the present invention is characterized by adding titanbicus flowers (including buds), water, brewing alcohol, and acetic acid bacteria to a container, and performing acetic acid fermentation while extracting quercetin glycosides from the titanbicus flowers (including buds) in a solution with an alcohol concentration of 1 v / v% to 15 v / v%.

[0013] The method is characterized in that the buds of titan bicus are used as the flowers (including buds) of titan bicus, and are added to the container in the form of buds.

[0014] The beverage of the present invention is characterized by containing the above-mentioned brewed vinegar. [Effects of the Invention]

[0015] The titanbicus brewed vinegar of the present invention is a brewed vinegar (hereinafter also referred to as titanbicus vinegar) obtained by acetic acid fermentation of titanbicus flowers (including buds). It has a quercetin glycoside concentration of 10 mg / 100 ml or more and an acidity of 4.0% or more in terms of acetic acid. This allows for the effective use of titanbicus flowers and buds while easily consuming quercetin glycoside. Furthermore, by producing the brewed vinegar, in addition to the pharmacological action of quercetin glycoside, the pharmacological action of acetic acid is expected to exert various physiological effects, such as an antihypertensive effect, as shown in the examples. Therefore, the titanbicus vinegar of the present invention is useful as a health-promoting product, for example, for use in sour seasonings and beverages.

[0016] The above-mentioned Titanicus vinegar contains isoquercitrin, rutin, and hyperoside, and among these, the concentration of isoquercitrin is the highest, so it is expected to have, for example, an effect of suppressing blood pressure rise due to ACE inhibitory activity.

[0017] The method for producing titanbicus vinegar of the present invention involves adding titanbicus flowers (including buds), water, brewing alcohol, and acetic acid bacteria to a container, and extracting quercetin glycoside from the titanbicus flowers (including buds) while performing acetic acid fermentation in a solution with an alcohol concentration of 1 v / v% to 15 v / v%.This allows extraction from the solid matter of titanbicus flowers (including buds) and acetic acid fermentation to proceed simultaneously, improving the production efficiency of titanbicus vinegar.

[0018] Since titanbicus buds are used as the flowers (including buds) and are added to the container in their intact form, acetic acid fermentation can be carried out without a grinding process to powder the buds, improving the production efficiency of titanbicus vinegar. Furthermore, as shown in the examples below, the concentration of quercetin glycosides, i.e., the content, can be increased more easily than when titanbicus powder is used for brewing. [Brief explanation of the drawings]

[0019] [Figure 1] FIG. 1 shows the structural formulas of quercetin and quercetin glycosides. [Figure 2] FIG. 1 shows the change in concentration of quercetin glycoside in Example 3. [Figure 3] FIG. 1 shows the correlation between isoquercitrin concentration and ACE inhibitory activity. [Figure 4] FIG. 1 shows the change in concentration of quercetin glycoside in Example 4. DETAILED DESCRIPTION OF THE INVENTION

[0020] The present inventors have conducted extensive research into brewed vinegar as a processed food in order to effectively utilize the flowers and buds of titan bicus. They have found that titan bicus vinegar has the function of inhibiting the activity of angiotensin-converting enzyme, and have also found that it is possible to enjoy the functionality of both acetic acid and the components contained in titan bicus flowers and buds. The present invention is based on these findings.

[0021] [Titan Bicus Vinegar] The titan bicus vinegar of the present invention uses titan bicus flowers (including buds) as a raw material.

[0022] Titanbicus (scientific name: Hibiscus x Titanbicus) is a selected hybrid between Hibiscus moscheutos and Hibiscus coccineus. Titanbicus varieties include Titan Leia (Ministry of Agriculture, Forestry and Fisheries Variety Registration No. 27541), Titan Flare (Ministry of Agriculture, Forestry and Fisheries Variety Registration No. 28201), Titan Pink (Ministry of Agriculture, Forestry and Fisheries Variety Registration No. 21812), Titan Peach White (Ministry of Agriculture, Forestry and Fisheries Variety Registration No. 21813), Titan Elf (Ministry of Agriculture, Forestry and Fisheries Variety Registration No. 27540), and Titan Pleiades. However, any variety can be used in the present invention.

[0023] In the Titan Bicus vinegar of the present invention, Titan Peach White (hereinafter simply referred to as Peach White) and Titan Pleiades (hereinafter simply referred to as Pleiades) are preferred because they are easier to increase the concentration of quercetin glycoside. Peach White has white flowers with a pale pink tint and a red eye in the center. Pleiades has pink flowers with a red eye in the center.

[0024] The titanbicus vinegar of the present invention contains components derived from titanbicus, specifically quercetin glycoside. Quercetin glycoside is a compound in which sugars are bound to the hydroxyl groups of quercetin, as shown in the following formula: In quercetin glycoside, the binding position and number of sugars are not particularly limited.

[0025] [ka]

[0026] The sugars in quercetin glycosides include monosaccharides, oligosaccharides, polysaccharides, etc. Monosaccharides include pentoses such as ribose and arabinose, and hexoses such as glucose, mannose, and galactose, and oligosaccharides and polysaccharides are formed by combining these monosaccharides. When the number of sugars is two or more, the sugars may be the same or different. Furthermore, the hydroxyl groups on the sugars may be substituted with other substituents, for example, modified into ester bonds or ether bonds.

[0027] The concentration of quercetin glycoside contained in titan bicus vinegar is, for example, 10 mg / 100 ml or more, preferably 30 mg / 100 ml or more, and may be 50 mg / 100 ml or more. A quercetin glycoside concentration above a predetermined value facilitates the expression of the pharmacological action of quercetin glycoside. In the present invention, the quercetin glycoside is preferably selected from the group consisting of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb. The upper limit of the quercetin glycoside concentration in titan bicus vinegar is not particularly limited, but is, for example, 200 mg / 100 ml.

[0028] Titanium bicus vinegar preferably contains isoquercitrin (quercetin with the monosaccharide glucose bound to the 3-position) as a quercetin glycoside, as shown in Figure 1. The concentration of isoquercitrin in Titanium bicus vinegar is, for example, 5.0 mg / 100 ml or more, preferably 10 mg / 100 ml or more, may be 20 mg / 100 ml or more, or may be 100 mg / 100 ml or less.

[0029] Titan bicus vinegar preferably contains two or more (more preferably three or more) types of quercetin glycosides. Specifically, in addition to isoquercitrin, it preferably contains rutin (quercetin with the disaccharide β-rutinose bound to the 3-position) and hyperoside (quercetin with the monosaccharide galactose bound to the 3-position). Furthermore, among these three components, titan bicus vinegar preferably has the highest concentration of isoquercitrin. In this case, the concentration of isoquercitrin may be, for example, 40% or more, or even 50% or more, of the total concentration of isoquercitrin, rutin, and hyperoside.

[0030] Titanium bicus vinegar preferably further contains Q3MG and Q3Samb as quercetin glycosides. In this case, the total concentration of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb in Titanium bicus vinegar is, for example, 10 mg / 100 ml or more, preferably 30 mg / 100 ml or more, and may be 50 mg / 100 ml or more. In this case, the concentration of isoquercitrin is, for example, 30% or more, or may be 40% or more, relative to the total concentration of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb. The concentration of Q3MG is, for example, 12% or less, or may be 10% or less, relative to the total concentration of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb.

[0031] The concentrations of each quercetin glycoside in Titanobicus vinegar can be measured by HPLC (high performance liquid chromatography) as described in the Examples.

[0032] As shown in Figure 1, all quercetin glycosides share the same quercetin structure, but differ in the type and number of sugars bound to them. For example, rutin is bound to a disaccharide, so it is not absorbed in the small intestine, but is absorbed in the large intestine after being metabolized by intestinal bacteria. On the other hand, isoquercitrin, which has one glucose bound to it, is absorbed in the small intestine. Hyperoside is also absorbed in the large intestine after being metabolized by intestinal bacteria. All quercetin glycosides are absorbed as free quercetin aglycones.

[0033] Titan Bicus Vinegar contains multiple quercetin glycosides (e.g., isoquercitrin, rutin, and hyperoside), which are thought to be effective in maintaining activity in the body, even though they share a common quercetin structure. Since isoquercitrin has also been reported to be more easily absorbed by the body than quercetin aglycone or rutin, a higher isoquercitrin content in Titan Bicus Vinegar is thought to be advantageous in terms of absorption.

[0034] Titanium bicus vinegar may contain quercetin glycosides other than those mentioned above. For example, it may contain other quercetin rutinosides, other quercetin glucosides, other quercetin galactosides, quercetin rhamnosides, quercetin glucuronides, etc. It may also contain quercetin.

[0035] The acidity of the titan bicus vinegar of the present invention is not particularly limited, but is preferably 4.0% or more, as specified for brewed vinegar in the vinegar quality labeling standards of the Japanese Agricultural Standards (JAS). The acidity can be calculated by the method specified in the Japanese Agricultural Standards for brewed vinegar. The upper limit of the acidity is not particularly limited, and is set to, for example, 15.0%. The preferred range of acidity is set to 4.0% to 8.0% in terms of acetic acid.

[0036] [How Titan Bicus Vinegar is Made] The raw materials for the titan bicus vinegar of the present invention are at least titan bicus flowers (including buds), water, brewer's alcohol, and acetic acid bacteria. By adding these raw materials to a container, a solution before brewing (hereinafter referred to as the undiluted solution) is obtained. Note that water, brewer's alcohol, and acetic acid bacteria may also be used as denatured alcohol that already contains these.

[0037] At least the flowers (including buds) are used as titan bicus. In this case, a mixture of flowers and buds may be used, or only the flowers or only the buds may be used. For example, non-standard buds or flowers that have already bloomed, which would have been discarded in the past, can be used as raw materials. The raw titan bicus may contain other parts such as leaves, seeds, stems, and roots, but it is preferable that these other parts are not included, as this would increase the concentration of ingredients. Furthermore, it is preferable that the calyx (sepals) are removed from the flowers and buds from the standpoint of hygiene, etc. In other words, it is preferable that the raw titan bicus does not contain calyx.

[0038] The form of titanbicus flowers (including buds) added to the container during brewing is not particularly limited, and they may be added in their original form (form A), powder form (form B), extract form (form C), etc.

[0039] In the case of Form A, flowers are added to a container while maintaining the shape of their petals, and buds are added to a container while maintaining the shape of the buds. In this case, the flowers and buds may be fresh or frozen, but frozen ones are preferably thawed to room temperature before being added.

[0040] In the case of Form B, the powder can be obtained, for example, by subjecting Form A to a drying step and a pulverization step. The drying method in the drying step is not particularly limited, and natural drying, heat drying, freeze drying, etc. can be used. In the case of heat drying, the heating temperature can be, for example, 40°C to 80°C. The pulverization method in the subsequent pulverization step is not particularly limited, and it can be carried out using a well-known pulverizer such as a mill, crusher, or stone mill. It is preferable to use a punching screen to make the particle size of the pulverized product somewhat uniform. The particle size and form of the obtained powder of titan bicus flowers and buds are not particularly limited. Note that after the pulverization step, an additional step (for example, a freeze-drying step or a sterilization step) may be added to obtain the powder.

[0041] In the case of Form C, the extract can be obtained by subjecting Form A or Form B to an extraction step. The extraction method used in the extraction step is not particularly limited, but examples include solvent extraction and supercritical extraction. Examples of extraction solvents include water; alcohols such as alcohols having 1 to 4 carbon atoms, such as methanol, ethanol, and isopropanol, and polyhydric alcohols, such as 1,3-butylene glycol, ethylene glycol, propylene glycol, and glycerin; ketones such as acetone and methyl ethyl ketone; esters such as ethyl acetate; hydrocarbons such as hexane; ethers such as diethyl ether; and acetonitrile. These extraction solvents may be used alone or in combination.

[0042] From the viewpoint of efficient extraction of quercetin glycosides, the extraction solvent is preferably water, alcohols, or a mixture thereof, with hydrous methanol and hydrous ethanol being more preferred. The alcohol concentration of the hydrous alcohol is, for example, 1% to 70% v / v, or may be 30% to 50% v / v. The extraction temperature is, for example, 20°C to 80°C, and methods such as immersion extraction, shaking extraction, and ultrasonic extraction can be used.

[0043] Among Forms A to C, it is preferable to use Form A or Form B, which are solid, from the viewpoint of the production efficiency of titanbicus vinegar. By using these, quercetin glycoside can be extracted from titanbicus flowers (including buds) in solution while acetic acid fermentation is performed, improving production efficiency. Furthermore, it is preferable to add the buds to the container as they are, as Form A, because this simplifies the production process and makes it easier to obtain quercetin glycoside at a high concentration.

[0044] The brewing alcohol raw material can be, for example, 99.5 v / v% alcohol, 95 v / v% alcohol, shochu, awamori, spirits, sake, etc. The brewing alcohol is added so that the alcohol concentration in the stock solution is, for example, 1 v / v% to 15 v / v%, preferably 1 v / v% to 10 v / v%, and more preferably 2 v / v% to 8 v / v.

[0045] The acetic acid bacteria used for acetic acid fermentation are not particularly limited, and for example, acetic acid bacteria belonging to the genus Acetobacter can be used. Specifically, Acetobacter aceti, Acetobacter pasteurianus, Acetobacter acetosum, etc. can be used. The acetic acid bacteria can be added in a predetermined amount to the stock solution, or can be added, for example, as a starter vinegar.

[0046] Examples of starter vinegar include a fermented liquid obtained by adding acetic acid bacteria to mash or purified alcohol refined from mash, followed by fermentation. This fermented liquid may be produced during the production process of brewed vinegar (defined in the "Vinegar Quality Labeling Standards" of the Japanese Agricultural Standards). Examples of brewed vinegar include grain vinegars such as rice vinegar, black rice vinegar, and black barley vinegar, and fruit vinegars such as apple vinegar and grape vinegar. When a fermented liquid derived from brewed vinegar is used as starter vinegar, the starter vinegar is added so that the acidity of the original liquid is, for example, 0.20% to 2.0%, preferably 0.20% to 1.5%. The addition of acetic acid bacteria and starter vinegar may be used in combination.

[0047] The raw materials for Titan Bicus Vinegar may further contain additives commonly used in acetic acid fermentation. For example, sake lees may also be used as a raw material. Sake lees are the solid matter remaining after pressing the mash of sake or other beverages. Because sake lees are rich in nutrients such as vitamins, adding sake lees is ideal for promoting health.

[0048] Acetic acid fermentation is carried out by heating the stock solution to which the above-mentioned ingredients have been added. The top of the container filled with the stock solution is covered with a lid (such as straw) that allows natural ventilation. In the case of static fermentation, for example, fermentation begins gradually from the top, which is exposed to air, due to the action of acetic acid bacteria. One to two days after fermentation, a bacterial film forms on the surface of the stock solution, and fermentation heat begins to be generated. During the fermentation period, the liquid temperature is preferably maintained at 30 to 45°C (for example, around 40°C). If necessary, the stock solution can be brought to around 40°C before fermentation begins. The vinegar, which has become heavier as a result of fermentation, sinks to the bottom of the container, creating natural convection that circulates within the container, allowing fermentation to proceed.

[0049] Acetic acid fermentation is carried out until the acidity of the solution exceeds 4.0%, and the fermentation period is approximately 30 to 50 days. After acetic acid fermentation is complete, the solution is filtered and sterilized as necessary to obtain Titanbicus vinegar.

[0050] The acetic acid fermentation method is preferably one that can maintain the liquid temperature at 30 to 45°C (for example, around 40°C) during the fermentation period. For example, acetic acid fermentation may be carried out with the container floating in a large-scale fermenter, or with the container attached to a thermostatic bath under temperature control, or with a heat insulating material wrapped around the container. The fermentation method is not limited to static fermentation, and well-known fermentation methods such as aeration fermentation can also be used.

[0051] The titan bicus vinegar of the present invention can be used as a sour seasoning containing acetic acid as a main component, and can also be used as a raw material for beverages intended to have health benefits.

[0052] [Beverage] The beverage of the present invention contains the titan bicus vinegar of the present invention described above, and is obtained by mixing the titan bicus vinegar into a beverage. When mixing the beverage, flavorings, vitamins, minerals, antioxidants, colorings, emulsifiers, preservatives, seasonings, sweeteners, fruit juice, honey, etc. may also be added.

[0053] This invention utilizes the flowers and buds of Titanobicus, which would normally be discarded in edible flower production, as processed foods. This process is significant as a process of upgrading by-products and waste into new products with higher quality and environmental value (upcycling). It also contributes to achieving Goal 12 of the Sustainable Development Goals (SDGs), "Responsible Consumption and Production." [Example]

[0054] [1. Study of extract] In the production of titanbicus vinegar, we first investigated the extract of titanbicus flowers (including buds). Because quercetin glycosides generally have low solubility in water, it is thought that higher ethanol concentrations facilitate their extraction. Therefore, the amount and extraction rate of quercetin glycosides extracted from peach white powder (peach white powder) were measured using extracts with ethanol concentrations of 0 v / v%, 5 v / v%, 10 v / v%, 20 v / v%, 25 v / v%, 35 v / v%, 50 v / v%, 75 v / v%, and 99.5 v / v%. The extraction rate for each ethanol solution was calculated relative to the extraction rate for 50 v / v% methanol, which was set at 100%.

[0055] (a) Manufacturing of peach white powder Peach white buds and flowers were used as raw materials. The peach white buds used as raw materials were harvested by cutting the base of the buds with the sepals still attached. In addition, blooming titan bicus flowers were also harvested with the sepals still attached. The harvested buds and flowers were promptly decanted and frozen for storage. The frozen buds and flowers were then subjected to a drying process (60°C, 24 hours), a crushing process (punching screen, size 0.4 mm), a freeze-drying process, and a sterilization process, resulting in powderization and sterilization. As an example, 7.89 kg of titan bicus powder was obtained from 94.5 kg of titan bicus buds (frozen product).

[0056] (b) Manufacturing of peach white powder extract Five ml of ethanol solution of various concentrations (0 v / v% to 99.5 v / v%) prepared with distilled water was added to a 15 ml centrifuge tube, and 0.127 g of peach white powder was added. After leaving the tube to stand at 40°C for 15 hours, the concentrations of various quercetin glycosides dissolved in the ethanol solution were measured under the following analytical conditions.

[0057] (c) Component analysis Detector: Photodiode array detector (measurement wavelength 254 nm) High performance liquid chromatography (HPLC): Agilent 1100 Series (Agilent) Column: ZORBAX Eclipse XDB-C18 (Agilent; column inner diameter 4.6 mm, column length 250 mm, particle size 5 μm) Column temperature: 40℃ Mobile phase A: acetonitrile (100%) Mobile phase B: 0.01% formic acid Flow rate: 1.000ml / min Delivery of each mobile phase: The mixing ratio of mobile phase A and mobile phase B was changed as shown in Table 1.

[0058] [Table 1]

[0059] Quantitative analysis of rutin, hyperoside, isoquercitrin, and quercetin was performed using calibration curves obtained from the respective standard substances (rutin trihydrate, hyperoside, isoquercitrin, and quercetin dihydrate). Each calibration curve was prepared by the HPLC analysis described above using a methanol solution (standard stock solution), a 10-fold diluted solution, and a 100-fold diluted solution of each standard substance. Since standard substances were unavailable for Q3Samb and Q3MG, the calibration curve for rutin was used for the quantification of Q3Samb, and the calibration curve for isoquercitrin was used for the quantification of Q3MG. Quantitative analysis using structurally similar compounds as model compounds is a commonly used method.

[0060] The detection times for quercetin and each quercetin glycoside under the above HPLC analysis conditions are as follows: Q3 Samb: Approx. 4.1 min Rutin: approx. 4.8 min Hyperoside: approx. 5.8 min Isoquercitrin: approx. 6.0 min Q3MG: Approx. 6.6 min Quercetin: Approximately 16.2 min

[0061] The extraction amount (concentration of quercetin glycoside) and extraction rate of each ethanol solution were calculated for each type of quercetin glycoside. The results are shown in Tables 2 and 3.

[0062] [Table 2]

[0063] [Table 3]

[0064] As shown in Table 2, when looking at the amount of each quercetin glycoside extracted, the amount (concentration) of isoquercitrin extracted was the highest, followed by hyperoside.

[0065] As shown in Table 3, the extraction rate of quercetin glycosides was generally good, at approximately 65%, at ethanol concentrations between 0% and 25% v / v. Because quercetin glycosides and quercetin are generally poorly soluble in water, this concentration would normally be considered insufficient for the extraction of various quercetin glycosides, but it was found that sufficient extraction was possible. At ethanol concentrations between 35% and 50% v / v, the extraction rate of quercetin glycosides was further improved. On the other hand, at higher concentrations (75% v / v and 99.5% v / v ethanol), the extraction rate of quercetin glycosides significantly decreased. Furthermore, isoquercitrin, the most abundant quercetin glycoside, showed the same tendency as the overall extraction rate of quercetin glycosides.

[0066] However, ethanol concentrations of 35% to 50% v / v, which are particularly effective in extracting quercetin glycosides, are not suitable for acetic acid fermentation by acetic acid bacteria. Therefore, extraction using such ethanol solutions requires subsequent dilution with water to adjust the ethanol concentration to a level suitable for acetic acid fermentation. However, for example, diluting a 35% to 50% v / v ethanol solution to 5% v / v reduces the quercetin glycoside concentration to approximately 1 / 10 to 1 / 6. This results in a decrease in the quercetin glycoside concentration, raising concerns about a loss of functionality due to the quercetin glycoside. Furthermore, this production method requires separate extraction and acetic acid fermentation steps, which increases time and labor costs.

[0067] On the other hand, when using an ethanol solution of 1 v / v% to 15 v / v%, for example, the amount of quercetin glycoside extracted is good, and acetic acid fermentation can be performed without dilution. Therefore, it is thought that it is possible to brew titanbicus vinegar with a higher concentration of quercetin glycoside than when extracting with a high-concentration ethanol solution. Furthermore, when using an ethanol solution of 1 v / v% to 15 v / v%, the extraction process and acetic acid fermentation process can be started simultaneously, which is thought to enable the production of titanbicus vinegar in a shorter time.

[0068] Therefore, the extraction rate of quercetin glycoside was calculated for other varieties (Leia, Pleiades) using a 5 v / v % ethanol solution according to the above procedure.

[0069] [Table 4]

[0070] As shown in Table 4, the extraction rate of quercetin glycoside was similar to that of Peach White powder when using Leia powder and Pleiades powder. Therefore, it is believed that, regardless of the variety of Titanobicus, it is possible to extract the quercetin glycoside contained in Titanobicus powder using, for example, a 5 v / v% ethanol solution, and that acetic acid fermentation can proceed simultaneously.

[0071] [2. Production of Titan Bicus Vinegar (Examples 1 to 3) using Titan Bicus Powder] The raw materials, 0.05 kg of peach white powder and 2 L of denatured alcohol, were placed in a plastic container. The denatured alcohol contains water, 95 v / v% brewer's alcohol, starter vinegar (a fermented product derived from brewer's vinegar), and sake lees, and has an alcohol concentration of approximately 5 v / v%.

[0072] The container containing the above ingredients was then floated in another fermentation tank (2 tons) for brewing. Due to the action of acetic acid bacteria contained in the starter vinegar, a film of bacteria formed on the surface of the liquid within a few days of brewing. In addition, the liquid temperature rose to approximately 40°C due to the heat of fermentation, and brewing was completed in 41 days.

[0073] The solution was sampled periodically from the start to the end of the brewing of Titan Bicus Vinegar. The collected samples were filtered through a 0.45 μm filter (filter material: hydrophilic mixed cellulose ester, diameter 25 mm, pore size: 0.45 μm, manufactured by Shimadzu GLC Corporation) and diluted 5-fold and 10-fold with ultrapure water to prepare the analytical sample. The concentration of quercetin glycoside was measured under the above analytical conditions. The results are shown in Table 5. The value on the 41st day of brewing is the value for Titan Bicus Vinegar.

[0074] The acidity of the obtained Titanbicus vinegar was 8.1%.

[0075] [Table 5]

[0076] As shown in Table 5, in the Titanicus vinegar of Example 1, which used peach white powder as a raw material, the quercetin glycoside concentration was greater than 30 mg / 100 ml, and the isoquercitrin concentration was greater than 10 mg / 100 ml. The percentages (%) of each quercetin glycoside relative to the total concentration of the five quercetin glycosides (Q3Samb, rutin, hyperoside, isoquercitrin, and Q3MG) and quercetin were 13.2% for Q3Samb, 15.8% for rutin, 23.3% for hyperoside, 35.6% for isoquercitrin, and 4.2% for Q3MG, resulting in a quercetin percentage (%) of 4.2%. Among these components, the isoquercitrin concentration was the highest. Furthermore, the isoquercitrin concentration was greater than 40% of the total concentration of isoquercitrin, rutin, and hyperoside.

[0077] On the other hand, the percentage of quercetin glycosides in peach white powder, calculated by analyzing the extract obtained by extracting peach white powder with 50 v / v% methanol, was 15.5% Q3Samb, 17.1% rutin, 23.0% hyperoside, 27.4% isoquercitrin, 11.0% Q3MG, and 6.1% quercetin. The percentage of quercetin glycosides contained in the above-mentioned Titan Bicus vinegar was roughly similar to the percentage in peach white powder itself.

[0078] Next, the Leia and Pleiades powders were used in the same brewing as the Peach White powder. The acidity of the resulting Titanbicus vinegar was 5.8% and 7.4%, respectively. The changes in the concentration of quercetin glycosides during brewing are shown in Tables 6 and 7.

[0079] [Table 6]

[0080] [Table 7]

[0081] As shown in Table 6, in the Titan bicus vinegar of Example 2, which used Leia powder as the raw material, the quercetin glycoside concentration was 20 mg / 100 ml or more, and the isoquercitrin concentration was 5 mg / 100 ml or more. Furthermore, as shown in Table 7, in the Titan bicus vinegar of Example 3, which used Pleiades powder as the raw material, the quercetin glycoside concentration was 30 mg / 100 ml or more, and the isoquercitrin concentration was 10 mg / 100 ml or more. Even when Leia powder or Pleiades powder was used, the isoquercitrin concentration was the highest among the five quercetin glycosides. Furthermore, the proportion of quercetin glycoside contained in each Titan bicus vinegar was similar to the proportion in each Titan bicus powder itself.

[0082] Figure 2 shows a graph of the change in quercetin glycoside concentration in Titanbicus vinegar of Example 3. As shown in Figure 2, the total concentration of quercetin glycoside in the solution increases with the number of brewing days. The concentrations of isoquercitrin, rutin, and hyperoside also increase in the solution. Figure 2 shows that acetic acid fermentation is progressing while quercetin glycoside is being extracted from Pleiades powder. Regarding Q3Samb and Q3MG, the concentrations initially increased, but then tended to decrease.

[0083] Thus, according to the above manufacturing method, the quercetin glycoside extraction process and the acetic acid fermentation process can be carried out simultaneously, and it has been found that the quercetin glycosides contained in titan bicus powder can be dissolved into brewed vinegar with almost no change in their content ratio.

[0084] [3. Angiotensin-converting enzyme inhibition test of Titan Bicus Vinegar in Examples 1 to 3] In this test, the dipeptide decomposed by ACE from the substrate (Hip-His-Leu) was made fluorescent using orthophthalaldehyde (OPA), and the ACE activity was determined by measuring the fluorescence intensity of the reaction product, and the effect of Titanbicus vinegar on ACE activity was evaluated.

[0085] Titanium bicus vinegar contains acetic acid due to acetic acid fermentation. Although acetic acid is known to lower blood pressure, in vitro evaluation of ACE inhibitory activity has reported that acetic acid does not exhibit ACE inhibitory activity. On the other hand, acetic acid is also known to affect the reaction by lowering the pH of the reaction solution when measuring ACE inhibitory activity.

[0086] Therefore, each titan bicus vinegar obtained in 2 above was neutralized to pH 8.2 with 30% sodium hydroxide solution, and then water was added to prepare a 900 mg / ml test solution. The test solution was diluted with HEPES buffer to prepare test solutions with the sample concentrations shown in Table 8.

[0087] 25 μl of the test solution prepared above was added to a 96-well microplate, followed by 25 μl of ACE solution, and the plate was left to stand at 37°C for 5 minutes. A control (untreated) was added with HEPES buffer instead of the test solution, and a blank (a phosphate buffer solution) was added instead of the ACE solution. After standing, the reaction was stopped by adding 25 μl of substrate solution, followed by 25 μl of OPA solution and the plate was left to stand at room temperature for 20 minutes. 25 μl of 0.1 mol / L hydrochloric acid was then added and the plate was left to stand at room temperature for 10 minutes.

[0088] The main reagents are as follows: HEPES buffer: Sodium chloride and HEPES (Sigma-Aldrich) were mixed with water to adjust the pH to 8.3, and then Triton-X was added to make a 0.1 mol / l solution with water (containing 0.3 mol / l sodium chloride and 0.01% Triton-X). ACE solution: A solution of ACE (derived from rabbit lung) [Sigma-Aldrich] at 13 mU / ml in phosphate buffer. Substrate solution: Hip-His-Leu (Peptide Institute, Inc.) at 8 mmol / L in HEPES buffer. OPA solution: A 1% solution of OPA (Fujifilm Wako Pure Chemical Industries, Ltd.) in methanol.

[0089] The fluorescence intensity of the reaction product was measured using a microplate reader [SpectraMax M2e Molecular Devices, LLC.] (excitation wavelength: 355 nm, emission wavelength: 460 nm).

[0090] The ACE activity and inhibition rate were calculated from the fluorescence intensity of each test solution relative to that of the untreated control using the following formula: Note that the fluorescence intensity of the blank was subtracted from each fluorescence intensity in the formula. ACE activity (%) = (fluorescence intensity of sample solution) / (fluorescence intensity of untreated control) × 100 ACE activity inhibition rate (%) = 100 - ACE activity (%)

[0091] The final concentration of the sample was calculated using the following formula. Final concentration (mg / ml) = sample concentration (mg / ml) x Sa / (Sa+Ace+Sub+NaOH+Opa+Hcl) = sample concentration (mg / ml) ÷ 6 Sa: Test solution dispensed volume (25 μl) Ace: ACE solution dispensed volume (25 μl) Sub: Substrate solution volume (25 μl) Naoh: 0.1 mol / l sodium hydroxide solution Dispense amount (25 μl) Opa: OPA solution dispensed volume (25 μl) Hcl: 0.1 mol / l hydrochloric acid (25 μl)

[0092] In addition, for each Titan Bicus vinegar, a logarithmic approximation curve was created from a graph with the ACE activity inhibition rate on the vertical axis and the sample concentration on the horizontal axis, and the IC 50 The 50% inhibitory concentration was calculated. The results are shown in Table 8.

[0093] [Table 8]

[0094] As shown in Table 8, ACE inhibitory activity was confirmed in all titan bicus vinegars. In this case, the acetic acid contained in titan bicus vinegar was neutralized, and the ACE inhibitory activity in titan bicus vinegar was determined to be the effect of components other than acetic acid (such as quercetin glycoside). Among the varieties, the ACE inhibitory activity of titan bicus vinegar in Example 3, which used Pleiades powder, was the highest, followed by Peach White powder and Leia powder. Furthermore, an analysis of the correlation between the concentration of isoquercitrin and ACE inhibitory activity in each titan bicus vinegar revealed an extremely high correlation (R 2 =0.9868) was observed (see Figure 3). Therefore, the flowers (including buds) of Titanobicus can be used to make vinegar, which has the same functionality of suppressing blood pressure elevation.

[0095] [4. Production of Titan Bicus Vinegar (Example 4) using Peach White Buds] Approximately 3.4 g of peach white buds and 11.22 L of denatured alcohol were added to a plastic container as raw materials. The denatured alcohol contained water, 95 v / v% brewer's alcohol, seed vinegar (a fermented product derived from brewer's vinegar), and sake lees, and was adjusted to an alcohol concentration of approximately 5 v / v%. The peach white buds were added to the container in their original form.

[0096] The amount of buds used is the amount of buds that is estimated to be sufficient to obtain the amount of titan bicus powder used in Examples 1 to 3, calculated backwards from an example in which titan bicus powder was obtained from titan bicus buds (7.89 kg of titan bicus powder was obtained from 94.5 kg of titan bicus buds (frozen)). In other words, the amount of components such as quercetin glycoside contained in the buds used in Example 4 is approximately the same as that of the titan bicus powder used in Examples 1 to 3.

[0097] The container containing the ingredients was then placed afloat in another fermentation tank and brewed in the same manner as in 2 above. The brewing was completed in 34 days, with the final acidity being 5.2%. The changes in quercetin glycoside concentration during brewing are shown in Table 9 and Figure 4.

[0098] [Table 9]

[0099] As shown in Table 9, the quercetin glycoside concentration in the Titanium Bicus vinegar of Example 4 was 50 mg / 100 ml or more, and the isoquercitrin concentration was 30 mg / 100 ml or more. The percentages (%) of each quercetin glycoside relative to the total concentration of the five quercetin glycosides (Q3Samb, rutin, hyperoside, isoquercitrin, and Q3MG) and quercetin were 19.5% for Q3Samb, 19.6% for rutin, 10.5% for hyperoside, 48.4% for isoquercitrin, and 2.0% for Q3MG, with the percentage of quercetin being 0.0%. Among these, the isoquercitrin concentration was the highest. Furthermore, the isoquercitrin concentration was 50% or more relative to the total concentration of isoquercitrin, rutin, and hyperoside.

[0100] Here, the concentration of quercetin glycoside in the Titan Bicus vinegar of Example 4, in which peach white buds were used as is, was 63.7 mg / 100 ml, which was approximately twice the concentration of quercetin glycoside (32.9 mg / 100 ml) in the Titan Bicus vinegar of Example 1, in which peach white powder was used, and more quercetin glycoside was able to be extracted.

[0101] [5. Angiotensin-converting enzyme inhibition test of Titan Bicus Vinegar] The ACE inhibitory activity was measured according to the procedure described above in 3. The results are shown in Table 10.

[0102] [Table 10]

[0103] As shown in Table 10, ACE inhibitory activity was confirmed in Titanobicus vinegar, which was prepared by using the buds as they were. 50 The value was 12.7 (50% inhibition at a final concentration of 12.7 mg / ml), indicating higher activity than in Example 1.

[0104] In this way, when titanbicus vinegar is brewed using the buds of titanbicus as they are, it is possible to extract more of the quercetin glycosides contained in the flowers (including the buds), and as a result, it is possible to produce a brewed vinegar with a higher quercetin glycoside balance.On the other hand, when titanbicus vinegar is brewed using titanbicus powder, it is possible to produce a brewed vinegar with a quercetin glycoside balance close to that contained in the flowers (including the buds). [Industrial Applicability]

[0105] The titan bicus vinegar of the present invention is a new processed food using titan bicus that is expected to exhibit physiological effects, and it can contribute to the effective use of resources and health promotion. Furthermore, the titan bicus vinegar can be said to be an upcycled product that adds new value to titan bicus flowers and buds that could not be shipped and were discarded during the production of edible flowers, etc., and can contribute to the achievement of the SDGs.

Claims

1. This is a brewed vinegar obtained by acetic acid fermentation of the flowers (including buds) of Titanobicus, characterized in that the concentration of quercetin glycoside is 10 mg / 100 ml or more and the acidity is 4.0% or more in terms of acetic acid.

2. The brewed vinegar of Titanobicus according to claim 1, characterized in that the brewed vinegar contains isoquercitrin, rutin, and hyperoside, with isoquercitrin having the highest concentration among these.

3. A method for producing brewed vinegar from titan bicus according to claim 1 or claim 2, A method for producing brewed vinegar from titanbicus, characterized by adding titanbicus flowers (including buds), water, brewing alcohol, and acetic acid bacteria to a container, and performing acetic acid fermentation while extracting quercetin glycoside from the titanbicus flowers (including buds) in a solution with an alcohol concentration of 1 v / v% to 15 v / v%.

4. A method for producing brewed vinegar from titan bicus according to claim 3, characterized in that the titan bicus flowers (including buds) are buds of titan bicus and are added to the container in the form of buds.

5. A beverage comprising the vinegar brewed from titan bicus according to claim 1 or 2.

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