Titan Bicus vinegar and its manufacturing method, and beverage

Abstract

The purpose of this study is to provide new processed foods using titan bicus, which is expected to have physiological effects, and to make effective use of resources. [Solution] Titanobicus brewed vinegar is made by acetic acid fermentation of Titanobicus flowers (including buds), and has a quercetin glycoside concentration of 10 mg / 100 ml or more and an acidity of 4.0% or more in acetic acid equivalent.It also contains isoquercitrin, rutin, and hyperoside, and of these, isoquercitrin has the highest concentration.

Description

[Technical field]

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

[0002] Conventionally, vinegar is known as vinegar, which is made by fermenting raw materials such as grains, fruits, and vegetables with acetic acid. As vinegar, mainly grain vinegars such as rice vinegar, rice black vinegar, and barley black vinegar, and fruit vinegars such as apple vinegar and grape vinegar are known. In recent years, various fruit vinegars are commercially available as health drinks because of their good taste. Acetic acid contained in vinegar is known to have effects such as fatigue recovery, lowering blood sugar levels, improving metabolism, and suppressing blood pressure rise. For example, there is a report that an acetic acid solution prepared to have the same acidity as barley black vinegar also suppressed blood pressure rise in rats, so acetic acid is thought to be one of the factors that suppresses blood pressure rise in vinegar.

[0003] On the other hand, among the various horticultural plants, Titan bicus is known as a plant that produces huge flowers. Titan bicus grows vigorously and is easy to grow, and is popular for its overwhelming presence. Titan bicus 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 that have been artificially bloomed from the buds by temperature treatment, are shipped to customers and then consumed.

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

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

[0006] [Patent Document 1] JP 2018-150262 A 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 that is expected to have physiological effects, and to achieve effective utilization of resources. [Means for solving the problem]

[0008] The titanic bicus brewed vinegar of the present invention is a brewed vinegar obtained by acetic acid fermentation of titanic bicus 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, "flowers of titan bicus (including buds)" is a concept that includes not only open flowers but also unopened flowers (buds), and does not limit the inclusion of actual buds. Furthermore, the form of "flowers of titan bicus (including buds)" is not particularly limited, and includes intact flowers and buds, as well as processed products thereof (e.g., powders) and extracts thereof; the same applies below.

[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 FIG. 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 titan bicus of the present invention is characterized by adding titan bicus flowers (including buds), water, brewing alcohol, and acetic acid bacteria to a container, and performing acetic acid fermentation while extracting quercetin glycoside from the titan bicus 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 flower (including bud) of titan bicus is a bud of titan bicus, and the flower is added to the container in the form of a bud.

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

[0015] The brewed vinegar of titan bicus of the present invention is a brewed vinegar (hereinafter also referred to as titan bicus vinegar) obtained by acetic acid fermentation of the flowers (including buds) of titan bicus, and 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, so that the quercetin glycoside can be easily ingested while making effective use of the flowers and buds of titan bicus. Furthermore, by making it into brewed vinegar, in addition to the pharmacological action of quercetin glycoside, various physiological actions such as a blood pressure rise suppression effect can be expected to be expressed due to the pharmacological action of acetic acid, as shown in the examples. Therefore, the titan bicus vinegar of the present invention is useful as a product for health promotion, for example, when used in sour seasonings and beverages.

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

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

[0019] [Figure 1] FIG. 1 shows the structural formulae of quercetin and quercetin glycosides. [Diagram 2] FIG. 1 shows the change in concentration of quercetin glycoside in Example 3. [Diagram 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 PREFERRED EMBODIMENTS

[0020] In order to effectively utilize the flowers and buds of Titan bicus, the present inventors have focused on brewed vinegar as a processed food and conducted extensive research. As a result, they have found that Titan bicus vinegar has the function of inhibiting the action of angiotensin converting enzyme, and further found that it is possible to enjoy both the functionality of acetic acid and the functionality of the components contained in the flowers and buds of Titan bicus. The present invention is based on such findings.

[0021] [Titan Bicas 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 of 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), Titan Pleiades, etc., but the present invention can be used without being limited to the variety.

[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 easy 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 titan bicus vinegar of the present invention contains a component derived from titan bicus, specifically, quercetin glycoside. Quercetin glycoside is a compound in which sugar is bound to the hydroxyl group of quercetin represented by 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. The 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. In addition, the hydroxyl group on the sugar may be substituted with another substituent, for example, modified to an ester bond or ether bond.

[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. When the concentration of quercetin glycoside is a predetermined value or more, the pharmacological action of quercetin glycoside is easily expressed. In the present invention, the quercetin glycoside is preferably composed of the group of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb. The upper limit of the concentration of quercetin glycoside in titan bicus vinegar is not particularly limited, but is, for example, 200 mg / 100 ml.

[0028] Titan bicus vinegar preferably contains isoquercitrin (a monosaccharide glucose bound to the 3-position of quercetin) as a quercetin glycoside, as shown in Figure 1. The concentration of isoquercitrin in Titan 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 as quercetin glycosides. Specifically, in addition to isoquercitrin, it preferably contains rutin (quercetin with β-rutinose, a disaccharide, bound to the 3rd position) and hyperoside (quercetin with galactose, a monosaccharide, bound to the 3rd position). Furthermore, among these three components, it is preferable that titan bicus vinegar has the highest concentration of isoquercitrin. In this case, the concentration of isoquercitrin may be, for example, 40% or more, or 50% or more, based on the total concentration of isoquercitrin, rutin, and hyperoside.

[0030] Titan 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 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. In this case, the concentration of isoquercitrin is, for example, 30% or more, and may be 40% or more, based on the total concentration of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb. The concentration of Q3MG is, for example, 12% or less, and may be 10% or less, based on the total concentration of isoquercitrin, rutin, hyperoside, Q3MG, and Q3Samb.

[0031] The concentration of each quercetin glycoside in Titan Bicus 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 metabolism by intestinal bacteria. On the other hand, isoquercitrin, which is bound to one glucose, is absorbed in the small intestine. Hyperoside is also absorbed in the large intestine after metabolism by intestinal bacteria. All quercetin glycosides are absorbed as free quercetin aglycone.

[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, since even if they have a common quercetin structure, there are differences in the place and time it takes to be absorbed into the body. In addition, it has been reported that isoquercitrin is more easily absorbed into the body than quercetin aglycone and rutin, so if Titan Bicus Vinegar contains more isoquercitrin, it is thought to be advantageous in terms of absorbability into the body.

[0034] Titan 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 standard 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%. As a preferred range, the acidity is set to 4.0% to 8.0% in terms of acetic acid.

[0036] [How Titan Bicas Vinegar is Made] The raw materials for the titan bicus vinegar of the present invention are at least the flowers (including buds) of titan bicus, 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 original solution) is obtained. Note that water, brewer's alcohol, and acetic acid bacteria may be used as denatured alcohol that already contains these.

[0037] As titan bicus, at least the flowers (including buds) are used. 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. Note that 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. Also, it is preferable that the calyx (selyx) is 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 the titanicus flowers (including buds) added to the container during brewing is not particularly limited, and they may be added in their original form (form A), in the form of a powder (form B), in the form of an extract (form C), or the like.

[0039] In the case of Form A, flowers are added to the container while maintaining the shape of, for example, petals, and buds are added to the container while maintaining the shape of buds. In this case, the flowers and buds may be in a fresh state or may be 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 carrying out a drying step and a pulverization step on Form A. The drying method in the drying step is not particularly limited, and can be natural drying, heat drying, freeze drying, etc. 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 can be carried out using a well-known pulverizer such as a mill, crusher, or stone mill. In order to make the particle size of the pulverized material somewhat uniform, it is preferable to use a punching screen. The particle size and shape of the obtained powder of the flowers and buds of Titan bicus are not particularly limited. After the pulverization step, another step (for example, a freeze-drying step or a sterilization step) may be further provided to obtain the powder.

[0041] In the case of Form C, the extract is obtained by carrying out an extraction process on Form A or Form B. The extraction method in the extraction process is not particularly limited, but examples thereof include solvent extraction and supercritical extraction. Examples of the extraction solvent 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 of two or more.

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

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

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

[0045] The acetic acid bacteria used in 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 to the stock solution in a predetermined amount, for example, as a seed vinegar.

[0046] As the seed vinegar, for example, a fermented liquid obtained by adding acetic acid bacteria to mash or purified alcohol refined from mash and fermenting it can be used. This fermented liquid may be one produced during the manufacturing process of brewed vinegar (specified in the "Vinegar Quality Labeling Standard" of the Japanese Agricultural Standards). Brewed vinegar includes grain vinegar such as rice vinegar, rice black vinegar, and barley black vinegar, and fruit vinegar such as apple vinegar and grape vinegar. When a fermented liquid derived from brewed vinegar is used as the seed vinegar, the seed 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%. In addition, the addition of acetic acid bacteria and the addition of seed vinegar may be used in combination.

[0047] The raw material for Titan Vikas Vinegar may further include additives commonly used in acetic acid fermentation. For example, sake lees may further be used as a raw material. Sake lees are solids that remain after squeezing the mash of sake and other products. Sake lees are rich in nutrients such as vitamins, so adding sake lees is ideal for promoting health.

[0048] The stock solution to which the above-mentioned raw materials have been added is heated to carry out acetic acid fermentation. The top of the container filled with the stock solution is covered with a lid material (such as straw) that can naturally draw in and out air. In the case of static fermentation, for example, fermentation starts little by little from the top exposed to air due to the action of acetic acid bacteria. A bacterial film forms on the surface of the stock solution 1 to 2 days after fermentation, 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 may be brought to around 40°C before fermentation is started. The vinegar, which has become heavier due to fermentation, sinks to the bottom of the container, creating natural convection, and fermentation progresses as it circulates inside the container.

[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 completed, the solution is filtered and sterilized as necessary to obtain Titanbicus vinegar.

[0050] The acetic acid fermentation method is preferably a method capable of maintaining the liquid temperature at 30 to 45°C (for example, around 40°C) during the fermentation period, and for example, acetic acid fermentation may be performed with the container floating in a large-scale fermenter, or with the container attached to a thermostatic tank under temperature control, or with a heat insulating material wrapped around the outer periphery of the container. The fermentation method is not limited to static fermentation, and well-known fermentation methods such as aeration fermentation can 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 that are 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 into a beverage, flavorings, vitamins, minerals, antioxidants, colorants, emulsifiers, preservatives, seasonings, sweeteners, fruit juice, honey, etc. can also be added.

[0053] This invention utilizes the flowers and buds of Titanobicus, which would normally be discarded in the production of edible flowers, as processed foods. Therefore, it is also meaningful as a process (upcycling) of upgrading by-products and waste to new products with higher quality and environmental value. It can also be said to contribute to the achievement of "12 Responsible Consumption and Production" of the Sustainable Development Goals (SDGs). EXAMPLES

[0054] [1. Study of the extract] In the production of Titanobicus vinegar, we first investigated the extract of Titanobicus flowers (including buds). Generally, quercetin glycoside has low solubility in water, so it is thought that the higher the ethanol concentration, the easier it is to extract quercetin glycoside. Therefore, the amount and extraction rate of quercetin glycoside extracted from Peach White powder (Peach White Powder) were measured using extracts with ethanol concentrations of 0v / v%, 5v / v%, 10v / v%, 20v / v%, 25v / v%, 35v / v%, 50v / v%, 75v / v%, and 99.5v / v% as follows. The extraction rate of each ethanol solution was calculated as a relative value when the extraction rate in 50v / v% methanol was set as 100%.

[0055] (a) Manufacture of peach white powder Peach white buds and flowers were used as raw materials. The peach white buds used as raw materials were harvested with the calyx still attached by cutting off the base of the bud. In addition, blooming titan bicus flowers were also harvested with the calyx 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 to powderize and sterilize them. 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 5 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 it 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 was performed for rutin, hyperoside, isoquercitrin, and quercetin using calibration curves obtained from the respective standard substances (rutin trihydrate, hyperoside, isoquercitrin, and quercetin dihydrate). Each calibration curve was prepared by the above HPLC analysis using a methanol solution (standard stock solution), a 10-fold diluted solution, and a 100-fold diluted solution of each standard substance. As it was not possible to obtain standard substances for Q3Samb and Q3MG, the calibration curve of rutin was used for the quantification of Q3Samb, and the calibration curve of isoquercitrin was used for the quantification of Q3MG. The method of quantitative analysis using substances with similar structures as model compounds is widely used.

[0060] The detection times for quercetin and each quercetin glycoside under the above HPLC analysis conditions are as follows: Q3Samb: 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: approx. 16.2 min

[0061] The amount of quercetin glycoside extracted from each ethanol solution (concentration of quercetin glycoside) and the extraction rate 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, at ethanol concentrations of 0 v / v% to 25 v / v%, the extraction rate of quercetin glycoside was good, at about 65%. Generally, quercetin glycoside and quercetin are poorly soluble in water, so this concentration would be considered insufficient to extract various quercetin glycosides, but it was found that they could be sufficiently extracted. At ethanol concentrations of 35 v / v% to 50 v / v%, the extraction rate of quercetin glycoside was further improved. On the other hand, at concentrations higher than this (75 v / v%, 99.5 v / v% ethanol), the extraction rate of quercetin glycoside was significantly reduced. In addition, isoquercitrin, which is the most abundantly extracted quercetin glycoside, showed the same tendency as the overall extraction rate of quercetin glycoside.

[0066] Incidentally, the ethanol concentration of 35v / v% to 50v / v%, which was particularly excellent in the extraction rate of quercetin glycoside, is not suitable for acetic acid fermentation by acetic acid bacteria. Therefore, when extraction is performed with an ethanol solution of such a concentration, it is necessary to dilute the ethanol with water to a concentration suitable for acetic acid fermentation. However, for example, when a 35v / v% to 50v / v% ethanol solution is diluted to 5v / v%, the concentration of the contained quercetin glycoside becomes about 1 / 10 to 1 / 6, and as a result, the concentration of the quercetin glycoside decreases, and there is a concern that the functionality of the quercetin glycoside may decrease. In addition, in such a production method, the extraction process and the acetic acid fermentation process must be performed separately, which requires time and labor.

[0067] On the other hand, when using an ethanol solution of 1v / v% to 15v / v%, for example, the amount of quercetin glycoside extracted is good, and acetic acid fermentation can be performed without dilution. Therefore, it is considered that it is possible to brew titan bicus vinegar with a higher concentration of quercetin glycoside than by extracting with a high concentration ethanol solution. In addition, if an ethanol solution of 1v / v% to 15v / v% is used, the extraction process and the acetic acid fermentation process can be started simultaneously, so it is considered that it is possible to produce titan bicus 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-mentioned 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 Titan bicus, it is possible to extract the quercetin glycoside contained in Titan bicus powder using, for example, a 5 v / v % ethanol solution, and that acetic acid fermentation can also proceed at the same time.

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

[0072] The container containing the above ingredients was brewed while floating in another fermentation tank (2 tons). Due to the action of acetic acid bacteria contained in the raw material, a film of bacteria formed on the surface of the liquid several days after brewing. In addition, the liquid temperature rose to about 40°C due to the heat of fermentation, and brewing was completed in 41 days.

[0073] From the start to the end of the brewing of Titan Bicus Vinegar, the solution was sampled periodically, and the collected sample was 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 Co., Ltd.), and diluted 5-fold and 10-fold with ultrapure water to prepare an 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 resulting titanbicus vinegar was 8.1%.

[0075] [Table 5]

[0076] As shown in Table 5, in the Titanbicus vinegar of Example 1 using peach white powder as a raw material, the concentration of quercetin glycoside was 30 mg / 100 ml or more, and the concentration of isoquercitrin was 10 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 13.2% for Q3Samb, 15.8% for rutin, 23.3% for hyperoside, 35.6% for isoquercitrin, and 4.2% for Q3MG, and the percentage (%) of quercetin was 4.2%. Among these components, the concentration of isoquercitrin was the highest. In addition, the concentration of isoquercitrin was 40% or more relative to the total concentration of isoquercitrin, rutin, and hyperoside.

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

[0078] Next, the Leia or Pleiades powder was used to brew in the same manner as the Peach White powder. The acidity of the resulting Titan Bicus vinegar was 5.8% and 7.4%, respectively. The changes in the concentration of quercetin glycoside during brewing are shown in Tables 6 and 7.

[0079] [Table 6]

[0080] [Table 7]

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

[0082] FIG. 2 shows a graph of the change in concentration of quercetin glycoside in Titan Bicus vinegar of Example 3. As shown in FIG. 2, the total concentration of quercetin glycoside in the solution increases with the number of brewing days. The concentrations of isoquercitrin, rutin, and hyperoside in the solution also increase. From FIG. 2, it can be seen that acetic acid fermentation is progressing while quercetin glycoside is extracted from the Pleiades powder. In addition, for Q3Samb and Q3MG, the concentrations initially increased, but then tended to decrease.

[0083] Thus, according to the above-mentioned manufacturing method, the quercetin glycoside extraction process and the acetic acid fermentation process can be carried out simultaneously, and it was 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 broken down by ACE from the substrate (Hip-His-Leu) was made fluorescent using ortho-phthalaldehyde (OPA), and the ACE activity was determined by measuring the fluorescence intensity of the reaction product, and the effect of Titan Bicus Vinegar on ACE activity was evaluated.

[0085] Titan Bicus vinegar contains acetic acid due to acetic acid fermentation. It is known that acetic acid lowers blood pressure, but in vitro ACE inhibitory activity evaluations have reported that acetic acid does not have ACE inhibitory activity. On the other hand, it is also known that acetic acid lowers the pH of the reaction solution when measuring ACE inhibitory activity, thereby affecting the reaction.

[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. The plate was tested in the same manner as above, with HEPES buffer added instead of the test solution as an untreated control, and with phosphate buffer added instead of the ACE solution as a blank. After leaving the plate to stand, 25 μl of substrate solution was added to stop the reaction, and 25 μl of OPA solution was added and the plate was left to stand at room temperature for 20 minutes. Furthermore, 25 μl of 0.1 mol / l hydrochloric acid was 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 and the resulting solution was adjusted to 0.1 mol / l with water (containing 0.3 mol / l sodium chloride and 0.01% Triton-X). ACE solution: 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, where each fluorescence intensity was a value obtained by subtracting the fluorescence intensity of the blank. ACE activity (%) = (fluorescence intensity of sample solution) / (fluorescence intensity of untreated control) x 100 ACE activity inhibition rate (%) = 100 - ACE activity (%)

[0091] The final concentration of the sample was calculated according to 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 volume (25 μl) Ace: Amount of ACE solution dispensed (25 μl) Sub: Substrate solution volume (25 μl) Naoh: 0.1 mol / l sodium hydroxide solution dispensed (25 μl) Opa: OPA solution dispensed volume (25 μl) Hcl: 0.1 mol / l hydrochloric acid dispensed (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 ingredients 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. In addition, analysis of the correlation between the concentration of isoquercitrin in each titan bicus vinegar and the ACE inhibitory activity revealed an extremely high correlation (R 2 = 0.9868) was observed (see Figure 3). Therefore, the flowers (including the 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] As raw materials, about 3.4g of peach white buds and 11.22L of denatured alcohol were added to a plastic container. The denatured alcohol contains water, 95v / v% brewer's alcohol, seed vinegar (a fermented product derived from brewer's vinegar), and sake lees, and the alcohol concentration is adjusted to about 5v / 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 enough to obtain the amount of titan bicus powder used in Examples 1 to 3 above, 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 above ingredients was brewed in a separate fermentation tank, floating it in the same manner as in 2. above. The brewing was completed in 34 days, and the final acidity was 5.2%. The change in the concentration of quercetin glycoside during brewing is shown in Table 9 and Figure 4.

[0098] [Table 9]

[0099] As shown in Table 9, in the titan bicus vinegar of Example 4, the concentration of quercetin glycoside was 50 mg / 100 ml or more, and the concentration of isoquercitrin 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, and the percentage (%) of quercetin was 0.0%. Among these, the concentration of isoquercitrin was the highest. In addition, the concentration of isoquercitrin 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 is approximately twice the concentration of quercetin glycoside in the Titan Bicus vinegar of Example 1, in which peach white powder was used (32.9 mg / 100 ml), and more quercetin glycoside was able to be dissolved.

[0101] [5. Angiotensin-converting enzyme inhibition test of Titan Bicas 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 Titanbicus vinegar, which was prepared by using the buds as is. 50 The value was 12.7 (inhibitory rate of 50% at a final concentration of 12.7 mg / ml), which showed higher activity than that of Example 1.

[0104] In this way, when titan bicus vinegar is brewed using titan bicus buds as is, it is possible to extract more of the quercetin glycoside contained in the flowers (including the buds), and as a result, it is possible to produce brewed vinegar that contains more quercetin glycoside. On the other hand, when titan bicus vinegar is brewed using titan bicus powder, it is possible to produce brewed vinegar with a balance of quercetin glycoside 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 have physiological effects, and can contribute to the effective use of resources and health promotion. In addition, the titan bicus vinegar can be said to be an upcycled product that adds new added value to the flowers and buds of titan bicus that were discarded during the production of edible flowers and cannot be shipped, and can contribute to the achievement of the SDGs.

Claims

1. A method for producing brewed vinegar from Titan bicus, the brewed vinegar 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; This method for producing brewed vinegar from titan bicus is characterized in that buds of titan bicus in their original bud form, water, brewing alcohol, and acetic acid bacteria are added to a container, and acetic acid fermentation is carried out while extracting quercetin glycoside from the titan bicus buds in a solution having an alcohol concentration of 1 v / v% to 15 v / v%.

2. The method for producing brewed vinegar from titan bicus according to claim 1, characterized in that the brewed vinegar contains isoquercitrin, rutin, and hyperoside, and among these, the concentration of isoquercitrin is the highest.

3. A method for producing a beverage containing brewed vinegar of titan bicus, comprising adding brewed vinegar of titan bicus produced by the method described in claim 1 or 2 to a beverage.

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