JP7807866B2 - beverage - Google Patents

Description

This technology relates to beverages.

Beverages made from fruit juice, such as fruit juice drinks and fruit juice-containing drinks, are among the beverages that many consumers prefer, and these beverages are required to have a good fruit juice flavor. However, beverages made from concentrated fruit juice may sometimes lack a fruit juice flavor.
For example, Patent Document 1 proposes a fruit juice enhancer containing γ-octadecalactone and/or δ-octadecalactone as an active ingredient, and discloses that the fruit juice enhancer can enhance the flavor of a fruit flavor or the fruit juice feel of a fruit-flavored food or drink, thereby improving palatability.

In recent years, there has been an increase in the number of beverages, such as mineral water, that contain small amounts of fruit juice or no fruit juice at all. Furthermore, in order to prevent precipitation due to fruit juice, beverages are sometimes made with a small amount of fruit juice or no fruit juice at all.

Japanese Patent Application Laid-Open No. 2019-97562

The main purpose of this technology is to provide beverages with an enhanced fruit juice flavor.

After extensive research, the inventors discovered that a beverage with an enhanced fruit juice flavor can be produced by combining four ingredients: a sweetener, a milk protein hydrolysate, a sour component, and a flavoring, and thus completed the present invention.

The present technology provides a beverage containing a sweetener, a milk protein hydrolysate, an acidic component, and a flavoring agent.
The present technology provides a method for producing a beverage, which includes a step of preparing a raw material liquid by mixing a sweetener, a milk protein hydrolysate, a sour component, and a flavoring.
The present technology provides a method for enhancing the fruit juice flavor of a beverage, which is characterized by adding a sweetener, a milk protein hydrolysate, a sour component, and a flavoring to the beverage.
The milk protein hydrolysate may be a casein protein hydrolysate.
The milk protein hydrolysate may be a milk protein hydrolysate containing Met-Lys-Pro.
The content of the milk protein hydrolysate may be 0.03% by mass or more.
The flavor may be a citrus flavor.
The beverage may be a soft drink or a milk-containing beverage.

This technology can provide beverages with an enhanced fruit juice flavor. Note that the effects described here are not necessarily limited to those described herein and may include any of the effects described in this technology.

Preferred embodiments for implementing the present technology are described below. Note that the embodiment described below is an example of a typical embodiment of the present technology, and should not be construed as narrowing the scope of the present technology. Note that percentages in this specification are expressed by mass unless otherwise specified. Furthermore, the upper and lower limits of each numerical range can be combined arbitrarily as desired.

<1. Beverages using the present technology>
This technology can provide a beverage containing a sweetener, a milk protein hydrolysate, a sour component, and a flavoring. This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener. Furthermore, this technology can improve the taste of the beverage.
In this technology, "fruit juice flavor" refers to the natural flavor, volume, taste, and complex, authentic fruit flavor that is reminiscent of actual fruit that can be found in high-fruit juice beverages.
In this technology, the "aftertaste of sweeteners" refers to the lingering sweetness, unpleasant taste, or other unpleasant sensations that remain on the tongue after drinking. Typical examples include those derived from high-intensity sweeteners. In addition, the use of high concentrations of sugar can also cause an unpleasant aftertaste. In this technology, no distinction is made based on the origin unless otherwise specified.

<1-1. (A) Sweetener>
The sweetener in the present technology means a component that can impart sweetness to food and drink (particularly beverages), and is a concept that includes sugars and components other than sugars that impart sweetness.
The sweetener used in the present technology may be any known sweetener that can be used in foods and beverages, or a commercially available product, and may be produced by a known production method.

Sweeteners include, but are not limited to, sugars, sugar alcohols, and high-intensity sweeteners; one or more selected from these groups can be used. Of these, sugars and/or high-intensity sweeteners are preferred. This technology can enhance the fruit juice flavor and/or improve the aftertaste of sweeteners.

The sugars may include monosaccharides and/or oligosaccharides, and sugar alcohols can be obtained by reducing these sugars. High-intensity sweeteners can also be classified into synthetic sweeteners, non-sugar natural sweeteners, amino acid-based sweeteners, etc.

The sweetener and its content can be measured by quantitative analysis of the sweetener using high performance liquid chromatography (HPLC).
In addition, a scale called sweetness intensity is used as an index of the sweetness of a sweetener. Sucrose is used as a standard substance, and sweetness is determined from the ratio of concentrations that show a sweetness intensity equivalent to that of sucrose at an arbitrary concentration, or the ratio to the threshold value of sucrose determined under the same conditions. For example, a certain sweet substance A is given, and the concentration of A that shows the same sweetness intensity as a 1% sucrose solution is measured by a sensory test. If the concentration of A at that time is 0.5%, the sweetness of sweet substance A is 2 compared to the sweetness of sucrose, which is 1. In this technology, the sweetness of a beverage to which no milk protein hydrolysate has been added is measured and used as the sweetness of the beverage of this technology.

The sugars are not particularly limited, but examples include sugar (e.g., white sugar, granulated sugar, brown sugar, brown sugar, etc.), starch syrup, powdered syrup, glucose, sugar mixture isomerized sugar, isomerized sugar, lactose, maltose, fructose, invert sugar, reduced malt syrup, honey, lactulose, trehalose, palatinose, D-xylose, fructooligosaccharides, maltooligosaccharides, etc., and one or more types selected from the group consisting of these can be used.

The sugar alcohols are not particularly limited, but include xylitol, sorbitol, maltitol, erythritol, etc., and one or more selected from the group consisting of these can be used.

High-intensity sweeteners are a general term for sweeteners with high sweetness.
The high-intensity sweetener is preferably a sweetener that is significantly sweeter than sucrose, specifically a sweetener that is 100 times or more sweeter than sucrose, based on the sweetness of sucrose.
For example, sucralose has a sweetness level of 600, acesulfame potassium has a sweetness level of 200, stevia has a sweetness level of 125, saccharin has a sweetness level of 350, aspartame has a sweetness level of 200, thaumatin has a sweetness level of 2500, and licorice extract has a sweetness level of 250 (Non-Patent Document 1: 2011, Vol. 106, No. 12, pp. 818-825, Journal of the Brewing Society of Japan, Basic Knowledge of Sweetness, Maebashi Kenji).

The high-intensity sweetener is not particularly limited, but examples thereof include sucralose, acesulfame potassium, saccharin sodium, cyclamate and its salts, thaumatin, aspartame, alitame, neotame, and stevioside contained in stevia extract, and one or more kinds selected from the group consisting of these can be used.
Of these, sucralose and/or acesulfame potassium are preferred, and of these, sucralose is more preferred, and a combined use of sucralose and acesulfame potassium is even more preferred.

<1-2. (B) Milk protein hydrolyzate>
The milk protein hydrolysate in the present technology is not particularly limited, but is preferably a hydrolysate of milk protein derived from milk such as cow's milk or skim milk powder.
Examples of the hydrolysis include acid hydrolysis, alkaline hydrolysis, and enzymatic hydrolysis. Among these, enzymatic hydrolysis is preferred from the viewpoint of ease of preparing various target peptide components. The enzymatic hydrolysis can be carried out under the enzymatic hydrolysis conditions described below.

Examples of the milk-derived protein include casein protein and whey protein, and one or more proteins selected from the group consisting of these can be used.
Casein proteins (hereinafter also referred to as "casein") can generally be classified into three types: α-casein, β-casein, and κ-casein. Whey proteins are also generally called milk serum proteins or soluble proteins.
Generally, whey proteins can be classified into serum albumin, β-lactalbumin, α-lactalbumin, immunoglobulin, proteose peptone, and the like.

In the present technology, a milk protein hydrolysate containing a peptide consisting of Met-Lys-Pro (SEQ ID NO: 1) (hereinafter also referred to as "tripeptide MKP") is preferred from the viewpoint of enhancing the fruit juice flavor and improving the aftertaste of sweeteners according to the present technology.
Furthermore, in the present technology, casein protein hydrolysate is preferred from the viewpoint of enhancing the fruit juice flavor and improving the aftertaste of sweeteners, and can also further improve the palatability of beverages.

[1-2-1. Casein protein hydrolysate]
The casein protein hydrolysate (hereinafter also referred to as "casein hydrolysate") used in the present technology will be described in detail below.
The casein hydrolysate used in the present technology is preferably a hydrolyzed product obtained by hydrolyzing casein protein derived from cow's milk, and contains various hydrolyzed components derived from casein protein.
The casein hydrolysate preferably satisfies at least one of the following conditions (a1) to (a4): (a1), (a2), (a3), and (a4), and more preferably satisfies two or more of the following conditions (a1), (a2), (a3), and (a4).
(a1) the average molecular weight of the casein hydrolysate is 1,200 daltons or less;
(a2) the decomposition rate of the casein hydrolysate is 20 to 30%;
(a3) the mass ratio of free amino acids to the total mass of all amino acids contained in the casein hydrolysate is 10% by mass or less;
(a4) Contains a peptide consisting of Met-Lys-Pro.

It is preferable that the casein hydrolysate contains at least the tripeptide MKP, from the viewpoint of better exerting the efficacy of the present technology.
Preferably, the casein hydrolysate contains the tripeptide MKP and further satisfies one or more of the conditions selected from the group consisting of (a1), (a2), and (a3).More preferably, the casein hydrolysate satisfies all of the conditions (a1) to (a4).

The tripeptide MKP of the present technology is known to have angiotensin-converting enzyme inhibitory activity, dipeptidyl peptidase-IV inhibitory activity, and other efficacies (e.g., Reference 1 (WO 2003/044044), Reference 2 (WO 2013/125622), Reference 3 (JP 2016-069343 A), etc.). However, it is not known to be involved in enhancing the fruit juice sensation or improving the aftertaste of sweeteners.

The content of tripeptide MKP in the casein hydrolysate is not particularly limited, but from the viewpoint of better exerting the effects of the present technology, the lower limit of the tripeptide MKP content is preferably 0.005% by mass or more, and more preferably 0.01% by mass or more, while from the viewpoint of hydrolysate production efficiency, the upper limit is preferably 0.2% by mass or less, and more preferably 0.1% by mass or less. The numerical range is even more preferably 0.01 to 0.1% by mass.

The method for producing casein hydrolysate used in this technology will be described in detail below.
[1-2-1(a) Raw materials for casein hydrolysate]
The raw material casein is primarily composed of milk-derived protein, and the casein is not particularly limited, but various commercially available caseins and caseinates can be used. More specifically, examples include casein lactate, casein sulfate, casein hydrochloride, sodium caseinate, potassium caseinate, calcium caseinate, magnesium caseinate, and any mixtures thereof. In addition, casein purified from cow's milk, skim milk, whole milk powder, or skim milk powder by a conventional method can also be used.

Casein hydrolysate, the active ingredient in this technology, is produced by hydrolyzing casein, a relatively inexpensive milk-derived biomaterial, and can therefore be produced stably, easily, and in large quantities.

[1-2-1(b) Method for producing casein hydrolysate]
The method for producing casein hydrolysate is not particularly limited, and examples include a method using an acid or alkali, a method using an enzyme such as a protease, etc. Among these, the method using an enzyme is preferred because it allows the hydrolysate to contain the desired peptides, etc.
A method for producing a casein hydrolysate using a protease will be specifically described below.

First, the raw material (casein) is dispersed and dissolved in water. There are no particular restrictions on the concentration of the solution, but a concentration of around 5-15% by mass (protein equivalent) is generally preferred for efficiency and ease of use.

Next, the pH of the solution is adjusted to about the optimum pH of the protease to be used, thereby preparing a raw material aqueous solution. Specifically, the pH of the solution is preferably adjusted to a pH of 7 to 10 using an alkaline solution, which is within the optimum pH range for many proteases.
The alkaline agent used for adjusting the pH is not particularly limited, but examples thereof include sodium hydroxide, potassium hydroxide, and potassium carbonate.

Next, a protease is added to the prepared raw material aqueous solution.
The protease may be derived from bacteria, animals, plants, or the like, and any of these may be used. One or more selected from the group consisting of these may also be used.

The bacterial proteolytic enzyme is not particularly limited, and examples thereof include endoproteases derived from the genus Bacillus, such as Alcalase (manufactured by Novozymes), Neutrase (manufactured by Novozymes), Protin A (manufactured by Yamato Chemical Industries, Ltd.), Protin P (manufactured by Yamato Chemical Industries, Ltd.), Proleather (manufactured by Amano Enzyme Co., Ltd.), Protease A (manufactured by Amano Enzyme Co., Ltd.), Protease N (manufactured by Amano Enzyme Co., Ltd.), Corolase 7089 (manufactured by Higuchi Shokai), Bioprase (manufactured by Nagase Chemtec Corporation), Orientase 90N (manufactured by HIBI Corporation), and Orientase 22BF (manufactured by HIBI Corporation).
The animal-derived proteolytic enzyme is not particularly limited, but examples thereof include PTN (manufactured by Novozymes) which contains trypsin as its main component, Trypsin V (manufactured by Nippon Biocon Co., Ltd.), and Pancreatin (manufactured by Amano Enzyme Inc.).
The plant-derived proteolytic enzyme is not particularly limited, but examples thereof include papain (manufactured by Amano Enzyme Co., Ltd.) and bromelain (manufactured by Amano Enzyme Co., Ltd.).
Furthermore, one or a combination of two or more selected from the group consisting of the above-mentioned proteolytic enzymes can also be used.

Protease is preferably dispersed and dissolved in cold water at 4-10°C before use. There are no particular restrictions on the concentration of the protease solution, but it is generally desirable to use an amount that results in an enzyme concentration of approximately 3-10% for reasons of efficiency and ease of use.

The amount of proteolytic enzyme used to hydrolyze casein varies depending on the substrate concentration, enzyme activity, reaction temperature, reaction time, etc., but a generally desirable ratio is 1,000 to 20,000 units (activity units) per gram of casein protein equivalent mass.

Protease activity units can be measured depending on the type of protease used.

When adding proteolytic enzymes, it is desirable to dissolve and add each type one at a time, but there are no particular restrictions on the order of addition.

During the enzymatic reaction, the temperature of the reaction system is not particularly limited and is selected from a practical range including the optimum temperature range for the expression of the enzyme activity, and is usually selected from the range of 30 to 60°C.
The reaction duration is difficult to determine in general terms because the progress varies depending on reaction conditions such as reaction temperature and initial pH, and there is a problem that, for example, if the reaction duration of the enzymatic reaction is set to a constant value, hydrolyzed products with different physicochemical properties may be produced in each production batch. Therefore, it is desirable to monitor the enzymatic reaction and determine the reaction duration so that the physicochemical properties of the casein hydrolysate reach desired values.
The enzyme reaction can be monitored, for example, by collecting a portion of the reaction solution and measuring the protein decomposition rate.

The enzymatic reaction is then stopped.
The enzymatic reaction is stopped by inactivating the enzyme in the hydrolyzed solution, which can be carried out by a conventional method, such as heat inactivation.
The conditions for the heat inactivation treatment (heating temperature, heating time, etc.) can be appropriately set to ensure sufficient inactivation, taking into account the thermal stability of the enzyme used. For example, the treatment can be carried out at a temperature range of 80 to 130°C for a holding time of 30 minutes to 2 seconds.

After the enzyme reaction has stopped, the resulting hydrolysis-deactivated liquid is preferably purified appropriately using one or more methods selected from the group consisting of (1) filtration, (2) membrane separation processes such as microfiltration and ultrafiltration, and (3) resin adsorption separation.

By carrying out the above-mentioned purification, it is possible to remove insoluble matter contained in the inactivated solution and reduce the amount of fat, lactose, and other unnecessary components. As a result, a casein hydrolysate can be obtained that is transparent in solution and does not become cloudy, precipitate, aggregate, or brown even when stored in solution for long periods of time, resulting in a casein hydrolysate with excellent storage stability.

In addition, the above-mentioned purification process can also improve the flavor of the casein hydrolysate used in this technology.

The filtration in (1) above can be carried out by a known method, for example, by using diatomaceous earth and a known device.
By carrying out filtration, insoluble matters that are present in the hydrolysis-inactivated solution and that have been produced during the hydrolysis reaction and/or during the enzyme heat-inactivation can be removed.

The membrane separation treatment (2) can be carried out using a known device. The known device is not particularly limited, but examples thereof include a microfiltration module and an ultrafiltration module, and more specifically, examples thereof include SEP1053 (manufactured by Asahi Kasei Corporation, molecular weight cutoff 3,000), SIP1053 (manufactured by Asahi Kasei Corporation, molecular weight cutoff 6,000), and SLP1053 (manufactured by Asahi Kasei Corporation, molecular weight cutoff 10,000).
In this case, a solution containing casein hydrolysate is obtained as a membrane permeate fraction after membrane separation treatment.
By carrying out the membrane separation treatment, insoluble matters that are present in the hydrolysis-inactivated solution and that have been produced during the hydrolysis reaction and/or during the enzyme thermal inactivation can be removed, as in the filtration in (1) above.

The resin adsorption separation in (3) can be carried out by a known method, for example, by filling a column with a resin and passing the hydrolysis/deactivation solution through the column. The resin is not particularly limited, and examples thereof include trade names such as Diaion, Sepabeads (manufactured by Mitsubishi Chemical Corporation), Amberlite XAD (manufactured by Organo Corporation), and KS-35 (manufactured by Ajinomoto Fine-Techno Co., Inc.).
Resin adsorption separation can be carried out continuously by filling a column with the resin and continuously introducing and discharging the hydrolysis and deactivation liquid into and from the column, or it can be carried out batchwise by adding a resin to the hydrolysis and deactivation liquid, allowing the resin to come into contact with the hydrolysis and deactivation liquid for a certain period of time, and then separating the hydrolysis and deactivation liquid from the resin.
The hydrolysis-inactivated solution may contain residual factors (e.g., peptides containing many hydrophobic amino acids) that can cause turbidity, precipitation, aggregation, browning, etc. during storage. These factors can be removed by performing resin adsorption separation.

After purification, the resulting solution containing the casein hydrolysate may also be sterilized.
The sterilization method may be a conventional heat treatment method or the like.
The heating temperature and holding time during the heat treatment may be appropriately set to conditions that allow sterilization. For example, sterilization can be achieved by heat treatment at 70 to 140° C. for 2 seconds to 30 minutes.
The heat sterilization method can be either a batch method or a continuous method, and in the continuous method, methods such as a plate heat exchange method, an infusion method, and an injection method can be used.

The resulting solution containing casein hydrolysate can be used as is, or, if necessary, the solution can be concentrated using a known method and used as a concentrate. The concentrate can also be dried using a known method and used as a powder.

In addition, after removing the insoluble components from the casein hydrolysate, a secondary hydrolysis may be performed by adding an endoprotease or exoprotease to improve flavor or physical properties.

Here, from the perspective of achieving better effects of the present technology, it is preferable to prepare the casein hydrolysate used in the present technology so that its average molecular weight is preferably 1,200 daltons or less, more preferably 1,000 daltons or less, even more preferably 800 daltons or less, and even more preferably 450 daltons or less. It is more preferable to prepare the casein hydrolysate so that its average molecular weight is more preferably 250 to 450 daltons, and even more preferably 360 to 390 daltons.

Furthermore, from the viewpoint of achieving better effects of the present technology, it is preferable to prepare the casein hydrolysate so that its hydrolysis rate is preferably 10% or more at the upper limit and 40% or less at the lower limit, and more preferably 20-30%.

In this technology, it is preferable to determine the degree of hydrolysis by determining reaction conditions such as reaction temperature and reaction duration so that the average molecular weight of the casein hydrolysate contained in the filtrate after removing insoluble matter generated during hydrolysis by filtration falls within the desired range, and/or so that the decomposition rate falls within the desired range.

Furthermore, from the viewpoint of achieving better effects of the present technology, it is more preferable to prepare the casein hydrolysate used in the present technology so that the proportion of the total mass of free amino acids to the total mass of all amino acids contained therein is preferably 15% by mass or less, and more preferably 10% by mass or less.

Furthermore, from the standpoint of the effectiveness and production efficiency of this technology, it is preferable to prepare the casein hydrolysate used in this technology so that the proportion of tripeptide MKP contained in the hydrolysate is preferably 0.001 to 1% by mass, more preferably 0.005 to 0.5% by mass, and even more preferably 0.01 to 0.1% by mass.

In this technology, the total mass ratio of free amino acids or the ratio of tripeptide MKP can be adjusted to the desired ratio by adjusting the type of enzyme used to hydrolyze casein, the amount of enzyme added, the reaction time, and/or the purification conditions after hydrolysis (membrane separation, resin adsorption separation), etc.

The following explains the amino acid decomposition rate, method for calculating average molecular weight, method for calculating amino acid release rate, and measurement of tripeptide MKP content in the casein hydrolysate of this technology.

<Method for calculating molecular weight>
The average molecular weight (Da: Dalton) of the casein hydrolysate in the present technology is determined based on the concept of number average molecular weight as follows.
The number average molecular weight (NAM) is a molecular weight that indicates the average value of the molecular weight of a polymer compound based on different indices as follows, as described in, for example, Non-Patent Document ("Fundamentals of Polymer Science," edited by the Society of Polymer Science, Inc., pp. 116-119, Tokyo Kagaku Dojin Co., Ltd., 1978):
That is, since high molecular weight compounds such as protein hydrolysates are heterogeneous substances and have a molecular weight distribution, the molecular weight of the protein hydrolysate must be expressed as an average molecular weight in order to handle it physicochemically. The number average molecular weight (hereinafter sometimes abbreviated as Mn) is the average for the number of molecules, and is defined by the following mathematical formula (1), where Mn is the molecular weight of peptide chain i and Ni is the number of molecules.

<Calculation method for decomposition rate>
The decomposition rate of the casein hydrolysate can be calculated using the following formula (2).

<Method for calculating the amino acid liberation rate>
In the present technology, the proportion of the total mass of free amino acids can be determined, for example, by the following procedure.

(i) Measurement of Amino Acid Composition For amino acids other than tryptophan, cysteine, and methionine, the sample was hydrolyzed with 6N hydrochloric acid at 110°C for 24 hours, tryptophan was subjected to alkaline decomposition with barium hydroxide at 110°C for 22 hours, and cysteine and methionine were treated with performic acid and then hydrolyzed with 6N hydrochloric acid at 110°C for 18 hours. Each was analyzed using an amino acid analyzer (Hitachi, Model 835) to measure the mass of the amino acid.
In this method, the amounts of glutamine and glutamic acid in a sample are quantified as the total amount of glutamine and glutamic acid, that is, the analytical value of glutamic acid.

(ii) Calculation of the total mass ratio of free amino acids Each amino acid composition in a sample is measured using the method described in (i) "Measuring Amino Acid Composition" above, and the mass of all amino acids in the sample is calculated by adding them up. Next, the sample is deproteinized with sulfosalicylic acid, and the mass of each remaining free amino acid is measured using the method described in (i) "Measuring Amino Acid Composition" above, and the mass of all free amino acids in the sample is calculated by adding them up. From these values, the total mass ratio of free amino acids in the sample is calculated using the following formula (3):

<Measurement of Tripeptide MKP Content>
(i) The sample powder is diluted and dissolved in 0.2% formic acid aqueous solution to a concentration of 1.0 mg/mL, and then ultrasonically crushed for 10 minutes. The powder solution is then filtered through a 0.22 μm PVDF filter (manufactured by Millipore) to prepare a powder solution, which is then analyzed by LC/MS under the following measurement conditions. Meanwhile, solutions of a chemically synthesized standard peptide of the tripeptide MKP (manufactured by Peptide Institute) at several concentrations are prepared, and LC/MS analysis is performed under the following measurement conditions to create a calibration curve.
Among the peaks obtained by analyzing the powder solution, those with the same molecular weight and retention time as the standard peptide are identified as having the same sequence as the standard peptide. The peak area of the standard peptide is compared with the peak area of the sample powder to determine the content of the tripeptide MKP in the powder solution.

(ii) MKP content (mg/g of casein hydrolysate)
MKP content (mg/g of casein hydrolysate) = [measured value of tripeptide MKP in the obtained casein hydrolysate (mg)] / [mass (g) of the obtained casein hydrolysate]
[Measured amount (mg) of tripeptide MKP in the obtained casein hydrolysate] is the measured amount of tripeptide MKP in the sample by "LC/MS" described below.

(iii) Instruments used in LC/MS Mass spectrometer: TSQ Quantum Discovery MAX (manufactured by Thermo Fisher Scientific).
High-performance liquid chromatograph: Prominence (Shimadzu Corporation), column: XBridge BEH300 C18 φ2.1 mm×250 mm, 3.5 μm (Waters).

(iv) LC/MS measurement conditions Mobile phase A: 0.2 wt % formic acid-aqueous solution Mobile phase B: 0.2 wt % formic acid-acetonitrile solution Time program: 2% B (0.0 min) - 25% B (5.0 min) - 65% B (5.1 min) - 65% B (10 min) - 85% B (10.1 min) - 85% B (13.0%) - 2% B (13.1 min) - STOP (30.0 min).
Sample injection volume: 10 μL, column temperature: 40°C, liquid flow rate: 200 μL/min
Analysis mode: SRM measurement.
Product Mass: m/z = 260.10 (Parent m/z = 375.21)

<1-3. (C) Sour Components>
In the present technology, the sour component means a component that can impart a sour taste to food and beverages (especially beverages), and is a concept that includes both acidic components and components other than acidic components that impart a sour taste, with components that can exhibit a sour taste in the mouth being preferred.
The sour component used in the present technology may be any known sour component that can be used in foods and beverages, or a commercially available product, and may be produced by a known production method.

The sour component is not particularly limited, but examples thereof include citric acid, malic acid, acetic acid, tartaric acid, glucono-delta-lactone, gluconic acid, phosphoric acid, ascorbic acid, phytic acid, lactic acid, fumaric acid, and salts thereof (e.g., sodium salts), and one or more selected from the group consisting of these can be used. Furthermore, organic acids having 1 to 2 or about 3 carbon atoms are preferred. This can enhance the fruit juice sensation and/or improve the aftertaste of the sweetener.
Among the sour components, citric acid or a salt thereof (e.g., trisodium salt) is preferred from the viewpoint of enhancing the fruit juice sensation and/or improving the aftertaste of sweeteners, and can also further improve the deliciousness of beverages. Examples of such salts include alkali metals (e.g., sodium, potassium, lithium, etc.) and alkaline earth metals (e.g., calcium, magnesium, etc.), and one or more selected from the group consisting of these can be used. Among these, alkali metals, and especially sodium, are preferred.

The sour component may also be a component contained in citrus fruits, such as citric acid contained in lemons or tartaric acid contained in grapes, and may also be a component contained in fruit juices.
Furthermore, citrus fruits are not particularly limited, but examples thereof include oranges, Unshu mandarins, grapefruits, lemons, limes, yuzu citrus, iyokan citrus, natsumikan citrus, hassaku citrus, ponkan citrus, shikuwasa citrus, and kabosu citrus, and one or more kinds selected from the group consisting of these can be used.

The acidity (%) can be measured and calculated as a citric acid equivalent in accordance with "2. Automatic titration" under "Measurement Methods" in the Japanese Agricultural Standards for Fruit Drinks (Last revised February 24, 2016, Ministry of Agriculture, Forestry and Fisheries Notification No. 489).

<1-4. (D) Fragrance>
The flavoring agent in the present technology means an ingredient that can impart a flavor to food and drink (particularly beverages), and any of naturally occurring ingredients, synthetic ingredients, and mixtures thereof can be used.
The flavoring agent used in the present technology may be any known flavoring agent that can be used in foods and beverages, or a commercially available product, and may be produced by a known production method.

The flavoring is not particularly limited, but fruit juice flavorings are preferred, such as citrus flavorings, apple flavorings, grape flavorings, strawberry flavorings, pineapple flavorings, banana flavorings, pear flavorings, peach flavorings, plum flavorings, blueberry flavorings, melon flavorings, guava flavorings, mango flavorings, acerola flavorings, and papaya flavorings, and one or more types selected from the group consisting of these can be used.
do.

Among these, the use of citrus flavorings is preferred because it can further enhance the citrus juice feeling, thereby improving the aftertaste of the sweetener and enhancing the deliciousness of the beverage.
Examples of the citrus flavor include lemon flavor, mandarin orange flavor, orange flavor, lime flavor, grapefruit flavor, and yuzu flavor. Among the citrus flavors, grapefruit flavor is preferred, and flavors containing limonene and/or citral are also preferred.

<1-5. Contents and mass ratios of (A) to (D)>
It is preferable to adjust the respective contents of the (A) sweetener, (B) milk protein hydrolysate, (C) sour component, and (D) flavoring in the beverage as follows:

<1-5-1. Sweetener content>
The content of the sweetener is not particularly limited, but it is preferable to blend the sweetener so that the sweetness of the beverage is 1 to 12, more preferably 2 to 12, even more preferably 4 to 11, and even more preferably 4 to 9. This can enhance the fruit juice sensation and/or improve the aftertaste of the sweetener.
When sugars are used as the sweetener, although there are no particular limitations, they are preferably contained in the beverage in an amount of 1 to 17% by mass, more preferably 2 to 16% by mass, and even more preferably 4 to 15% by mass, which can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener.

When a high-intensity sweetener is used as the sweetener, although there are no particular limitations, it is preferable to add it to the beverage at a concentration of preferably 0.005 to 0.06% by mass, more preferably 0.01 to 0.05% by mass, and even more preferably 0.02 to 0.045% by mass. This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener.

<1-5-2. Content of milk protein hydrolysate>
The content of the milk protein hydrolysate in the beverage is not particularly limited, but the lower limit is preferably 0.03% by mass or more, more preferably 0.04% by mass or more, even more preferably 0.05% by mass or more, and the upper limit is preferably 2% by mass or less, more preferably 1.5% by mass or less, even more preferably 0.5% by mass or less, and even more preferably 0.4% by mass or less. The preferred range of the content of the milk protein hydrolysate in the beverage is more preferably 0.05 to 0.5% by mass. This can further enhance the fruit juice flavor and/or improve the aftertaste of sweeteners. The present technology has the excellent advantage that a small amount of milk protein hydrolysate can enhance the fruit juice flavor even in cases where there is no fruit juice or low fruit juice.
In order to enhance the fruit juice flavor and improve the aftertaste of sweeteners in soft drinks, the amount is more preferably 0.05 to 0.2% by mass, and even more preferably 0.05 to 0.075% by mass.
In order to enhance the fruit juice flavor and improve the aftertaste of sweeteners in milk-containing beverages, the amount is more preferably 0.3 to 0.5% by mass, and even more preferably 0.3 to 0.4% by mass.

<Tripeptide MKP content>
In the present technology, it is preferable that the (a4) tripeptide MKP is contained in the beverage, which can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and can also improve the deliciousness of the beverage.
The content of (a4) tripeptide MKP in a beverage is not particularly limited, but the lower limit is preferably 0.00002% by mass or more, more preferably 0.00009% by mass or more, even more preferably 0.0001% by mass or more, and the upper limit is preferably 0.001% by mass or less, more preferably 0.0008% by mass or less, and even more preferably 0.0005% by mass or less. The preferred range of the content of (a4) tripeptide MKP in a beverage is more preferably 0.00002 to 0.001% by mass, even more preferably 0.00009 to 0.0008% by mass, and even more preferably 0.0001 to 0.0005% by mass. This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and can further improve the deliciousness of the beverage.

<1-5-3. Content of sour components>
The sour component is not particularly limited, but is preferably contained in the beverage in an amount of 0.05 to 0.5% by mass, more preferably 0.1 to 0.5% by mass.
The acidity (%) of the beverage is preferably 0.01 to 1.0, more preferably 0.05 to 0.5, and even more preferably 0.1 to 0.4, thereby enabling the fruit juice flavor to be enhanced and/or the aftertaste of the sweetener to be improved.

<1-5-4. Fragrance content>
The flavoring is not particularly limited, but is preferably contained in the beverage in an amount of 0.5% by mass or less, more preferably 0.01 to 0.3% by mass, which can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener.

<1-5-5. Content of milk protein hydrolysate relative to sweetness level 1>
The lower limit of the content of the milk protein hydrolysate to be blended into the beverage is preferably 0.004% by mass or more, more preferably 0.006% by mass or more, and the upper limit is preferably 0.1% by mass or less, more preferably 0.08% by mass or less, even more preferably 0.06% by mass or less, still more preferably 0.05% by mass or less, and more preferably 0.04% by mass or less, relative to the sweetness level of the beverage of 1. The range of the milk protein hydrolysate content in the beverage relative to the sweetness level of the beverage of 1 is more preferably 0.004 to 0.1% by mass, even more preferably 0.008 to 0.08% by mass, and even more preferably 0.009 to 0.06% by mass.
Furthermore, relative to a sweetness level of 1 for a beverage, the content of milk protein hydrolysate in the beverage is more preferably 0.005 to 0.015 mass% for soft drinks, and more preferably 0.025 to 0.04 mass% for milk-containing beverages.
This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and can also improve the deliciousness of the beverage. Furthermore, this technology has the excellent advantage of being able to enhance the fruit juice flavor even in cases where there is no fruit juice or a low amount of fruit juice.

<1-5-6. Content of milk protein hydrolysate per [sweetness/acidity] ratio of 1>
The sweetness/acidity ratio of the beverage is not particularly limited, but is preferably 5 to 80, more preferably 10 to 60. Furthermore, the sweetness/acidity ratio of the beverage is more preferably 15 to 30 for soft drinks, and more preferably 40 to 60 for milk-containing beverages. This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and can also improve the deliciousness of the beverage. Furthermore, this technology has the excellent advantage of being able to enhance the fruit juice flavor even in beverages containing no or low amounts of fruit juice.

The content of milk protein hydrolysate blended into a beverage is preferably 0.0015% by mass or more, more preferably 0.002% by mass or more, and even more preferably 0.0025% by mass or more, relative to the beverage's sweetness/acidity ratio of 1. The upper limit is preferably 0.015% by mass or less, more preferably 0.0125% by mass or less, and even more preferably 0.01% by mass or less. For soft drinks, a content of 0.0025 to 0.00375% by mass is more preferred, relative to the beverage's sweetness/acidity ratio of 1. For milk-containing beverages, a content of 0.006 to 0.008% by mass is more preferred. This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and also improve the palatability of the beverage. This can also provide a beverage with a fruit juice flavor, even in beverages that contain no or low amounts of fruit juice.

<1-5-7. Mass ratio of sour component content and milk protein hydrolysate content>
The mass ratio of the sour component content to the milk protein hydrolysate content in the beverage is not particularly limited, but is preferably 10 to 0.1, more preferably 6 to 0.5. This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and can also improve the deliciousness of the beverage. This can also provide a beverage with a fruit juice flavor, even if it contains no or little fruit juice.
In the case of soft drinks, the mass ratio of [content of sour components/content of milk protein hydrolysate] in the beverage is preferably 10 to 0.5, more preferably 6 to 1.5, and even more preferably 6 to 4, from the viewpoints of enhancing the fruit juice flavor, improving the aftertaste of sweeteners, and improving the deliciousness of the beverage.
In the case of milk-containing beverages, the mass ratio of [content of sour components/content of milk protein hydrolysate] in the beverage is preferably 5 to 0.1, more preferably 3 to 0.2, even more preferably 2.5 to 0.4, and even more preferably 1 to 0.5, from the viewpoints of enhancing the fruit juice flavor, improving the aftertaste of sweeteners, and improving the deliciousness of the beverage.

<1-5-8. Mass ratio of flavoring content and milk protein hydrolysate content>
The mass ratio of [flavoring content/milk protein hydrolysate content] in the beverage is not particularly limited, but is preferably 3 to 0.1, more preferably 2 to 0.1, and even more preferably 1.5 to 0.2. This can enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and can also improve the deliciousness of the beverage. This can also provide a beverage with a fruit juice flavor, even if it contains no or little fruit juice.
In the case of soft drinks, the mass ratio of [flavoring content/milk protein hydrolysate content] in the beverage is preferably 1.2 to 0.3, more preferably 1.2 to 0.6, and even more preferably 1.2 to 0.8, from the viewpoints of enhancing the fruit juice flavor, improving the aftertaste of the sweetener, and improving the deliciousness of the beverage.
In the case of a milk-containing beverage, the mass ratio of [flavoring content/milk protein hydrolysate content] in the beverage is preferably 1.1 to 0.2, more preferably 0.3 to 0.2, and even more preferably 0.35 to 0.25, from the viewpoints of enhancing the fruit juice flavor, improving the aftertaste of the sweetener, and improving the deliciousness of the beverage.

<1-6. Optional ingredients＞
The beverage of the present technology may contain optional ingredients that can be used as ingredients in foods, beverages, etc., as needed, within the scope that does not impair the effects of the present technology.
Examples of optional ingredients include acidic ingredients, probiotics, milk ingredients, dietary fiber, human milk oligosaccharides, stabilizers, flavoring ingredients other than the (D) flavoring, vegetable oils and fats, vegetable milk, thickening polysaccharides, oils and fats, proteins, amino acids, organic acids, vitamins, and inorganic salts, and one or more selected from the group consisting of these can be used.

The acidic component may be the sour component described above, but may also be used as a pH adjuster, and the acidic component can impart a sour taste to the composition of the present technology or place it in the acidic range.
The acidic component is not particularly limited, but in addition to the above-mentioned examples of sour components, examples include gluconic acid, phytic acid, phosphoric acid, carbon dioxide, and salts thereof, and one or more selected from the group consisting of these can be used.

Probiotics are not particularly limited, but examples include lactic acid bacteria, Bifidobacterium bacteria, acetic acid bacteria, and Bacillus subtilis, and one or more species selected from the group consisting of these can be used. Of these, lactic acid bacteria and/or Bifidobacterium bacteria (e.g., B. longum, B. infantis, B. breve, etc.) are preferred. Probiotics may be live bacteria, killed bacteria, or cultures using these, but live bacteria are preferred from the perspective of probiotic effect. It is also possible to expect probiotic effects by blending bacteria, bacterial cultures, fermented products, or fermented milk.

The milk component may contain at least milk protein and/or milk fat. Fermented milk obtained by fermenting the milk component with a fermenting bacterium (such as the probiotics mentioned above) may also be used as the milk component. Fermented milk can be obtained by fermentation using a general fermentation method, but the fermentation method is not particularly limited.

The milk components that can be used are mainly those derived from mammalian milk (for example, cow's milk, goat's milk, sheep's milk, horse's milk, etc.), preferably those derived from cow's milk.
Examples of the milk component include raw milk, cow's milk, skim milk, partially skim milk, concentrated milk, concentrated skim milk, condensed milk, whole milk powder, skim milk powder, whey, whey protein concentrate (WPC), whey protein isolate (WPI), whole milk protein concentrate (TMP), cream, cream powder, and whey powder, and one or more selected from the group consisting of these can be used.

As dietary fiber, sugars that are indigestible by human enzymes are suitable, and examples of such sugars include polysaccharides and/or oligosaccharides. In this specification, "indigestible" means that the sugars are difficult to digest by human digestive enzymes.
The sugars may be, but are not limited to, plant-derived and/or bacterial-derived dietary fibers. Dietary fibers that can be assimilated by Bifidobacterium bacteria and/or lactic acid bacteria are preferred. Water-soluble dietary fibers are preferably polysaccharides and/or oligosaccharides that are soluble in water at about 4 to 30°C.

Examples of dietary fiber in the present technology include oligosaccharides (e.g., galactooligosaccharides, fructooligosaccharides, soybean oligosaccharides, xylooligosaccharides, isomaltooligosaccharides, raffinose, lactulose, coffee bean mannooligosaccharides, gluconic acid, etc.) and dietary fibers (polydextrose, inulin, xylan, arabinan, pectin, galactan, cellulose, soybean fiber, dextrin, dextran, resistant dextrin, etc.), and one or more types selected from the group consisting of these can be used. The dietary fiber content is not particularly limited, but can be, for example, 0.5 to 3% by mass.

For example, resistant dextrin, a type of dietary fiber, is a water-soluble dietary fiber prepared from starch. This resistant dextrin can be obtained, for example, by enzymatically digesting roasted dextrin, or by hydrogenating the resulting product after enzymatic digestion. It can also be a commercially available product such as Fibersol 2 (manufactured by Matsutani Chemical Industry Co., Ltd.).

Examples of human milk oligosaccharides include 2'-fucosyllactose, 3-fucosyllactose, 2',3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-neofucopentaose, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, and lacto-N-neofucopentaose. Examples include neutral human milk oligosaccharides such as lactose V, lacto-N-difucohexaose I, lacto-N-difucohexaose II, 6'-galactosyllactose, 3'-galactosyllactose, lacto-N-hexaose, and lacto-N-neohexaose; and acidic human milk oligosaccharides such as 3'-sialyllactose, 6'-sialyllactose, 3-fucosyl-3'-sialyllactose, and disialyl-lacto-N-tetraose. One or more types selected from the group consisting of these can be used.

The stabilizer is not particularly limited, but examples include high methoxyl pectin, sodium carboxymethylcellulose, and soybean polysaccharides, and one or more selected from the group consisting of these can be used. Soybean polysaccharides are polysaccharides obtained from soybeans, and their main component is hemicellulose. Commercially available products may also be used, and examples of commercially available products include, but are not limited to, high methoxyl pectin (SM-666, manufactured by San-Ei Gen F.F.I.), sodium carboxymethylcellulose (Cellogen FZ (product name), manufactured by Dai-ichi Kogyo Seiyaku Co., Ltd.), and soybean polysaccharides (SM-1200, manufactured by San-Ei Gen F.F.I.).

The flavoring component other than the (D) flavoring is not particularly limited, but any component that can be used for the purpose of flavoring and aromatizing beverages can be used. In addition to the examples of the (D) flavoring, examples of flavoring components include coffee, teas (e.g., black tea, green tea, roasted green tea, bancha tea, sencha tea, oolong tea, etc.) and extracts thereof; vegetable juices (e.g., tomato, carrot, etc.), powders thereof, or flavors thereof; and one or more selected from the group consisting of these can be used.

<1-7. Providing beverages using this technology>

The beverages of the present technology are not particularly limited, but examples thereof include carbonated beverages, natural fruit juices, fruit juice drinks, soft drinks containing fruit juice, fruit pulp drinks, fruit drinks containing fruit pieces, vegetable-based beverages, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered beverages, concentrated beverages, sports drinks, nutritional drinks, alcoholic beverages, and other beverages, and the beverages of the present technology may be one or more selected from these groups.
The beverage of the present technology may be an acidic beverage. The acidic beverage generally refers to a beverage having a pH of less than 7. The pH (20°C) of the beverage is preferably 6 or less, more preferably 2 to 6, and even more preferably 2 to 4.

Common beverages include, for example, soft drinks (with an alcohol content of less than 1%) and alcoholic beverages, and beverages that do not contain alcohol (specifically, beverages with an alcohol content of less than 1%) are usually referred to as soft drinks.
The beverage of the present technology is preferably a soft drink or a milk-containing beverage, and is preferably a beverage with an alcohol content of less than 1%.

<1-7-1. Soft drinks＞
The "soft drink" of the present technology is, for example, one or more types selected from the group consisting of carbonated drinks, fruit juice drinks, vegetable juices, sports drinks, tea drinks, coffee drinks, dairy drinks, fermented milk drinks, lactic acid bacteria drinks, etc. In the present technology, soft drinks with no fruit juice or low fruit juice content are preferred.

<1-7-2. Milk-containing beverages＞
The "milk-containing beverage" of the present technology is a beverage containing at least a milk component, and may be classified as a milk beverage, a soft drink, or the like. The milk component used in the present technology may contain at least one of milk-derived components, and may contain milk protein and/or milk fat and/or lactose and/or milk-derived minerals. The milk component may be either fermented milk or non-fermented milk. In the present technology, milk-containing beverages with no or low fruit juice content are preferred.

In the milk-containing beverage, the above-mentioned "milk components" can be used as appropriate, and the contents of non-fat milk solids and milk fat in the beverage can be adjusted as appropriate.
When adjusting the milk fat content in a beverage, in addition to the above-mentioned "milk components," a "milk fat source" can be used, and the milk fat source is not particularly limited as long as it contains milk fat. Examples of the milk fat source include cream, butter, butter oil, and cream cheese, and one or more selected from the group consisting of these can be used.

<Non-fat milk solids>
In a milk-containing beverage, the content of non-fat milk solids in the beverage is not particularly limited, but the lower limit is preferably 0.1% by mass or more, more preferably 0.5% by mass or more, and the upper limit is preferably 12.0% by mass or less, more preferably 8.0% by mass or less. The numerical range is more preferably 0.1 to 12.0% by mass, and even more preferably 0.5 to 8.0% by mass. This is satisfactory.

<Milk fat content>
In a milk-containing beverage, the milk fat content in the beverage is not particularly limited, but the upper limit of the milk fat content is preferably 4.0% by mass or less, and more preferably 3.5% by mass or less.
When the beverage of the present technology is low-fat, the upper limit of the milk fat content in the beverage is preferably 1.5% by mass or less, more preferably 1.2% by mass or less, even more preferably 1.0% by mass or less, and even more preferably 0.8% by mass or less.

In addition, in this technology, "milk solids" refers to the sum of non-fat milk solids and milk fat. The contents of milk solids, non-fat milk solids, milk fat, and other milk components in this technology can be measured using the quantitative methods described in the "Testing Methods for Compositional Standards of Milk, etc." in the "Ministerial Ordinance on Milk, etc. (Ministerial Ordinance on Compositional Standards, etc. of Milk and Dairy Products)."

<1-7-3. Low fruit juice drinks, non-fruit juice drinks＞
With this technology, it is possible to impart or enhance the fruit juice feel even to beverages with a low fruit juice content (e.g., low-fruit juice beverages) or beverages that do not contain fruit juice (so-called juice-free beverages, e.g., less than 5% fruit juice). Examples of beverages with a low fruit juice content include beverages with less than 10% fruit juice and beverages with 5% to less than 10% fruit juice. Examples of beverages that do not contain fruit juice (so-called juice-free beverages) include beverages with less than 5% fruit juice, beverages with 4%, 3%, 2%, or 1% or less fruit juice, and beverages with no added fruit juice.
The definition of "no fruit juice" in the "no fruit juice beverage" of this technology is less than 5% fruit juice, based on the Consumer Affairs Agency of Japan's "Labeling for juice-free soft drinks, etc." (https://www.caa.go.jp/policies/policy/representation/fair_labeling/representation_regulation/case_006/). The Consumer Affairs Agency of Japan defines "etc." in "no fruit juice soft drinks, etc." as meaning milk drinks, fermented milk, lactic acid bacteria drinks, powdered drinks, ice cream, and other products that are in containers or packaging, including soft drinks.

The beverage of the present technology can also be applied to food and beverage products, pharmaceutical products, etc.
The present technology may be used on humans or non-human animals (e.g., pets, livestock, etc.), and may be used for therapeutic or non-therapeutic purposes. "Non-therapeutic purposes" is a concept that does not include medical procedures, i.e., treatment of the human body through therapy.

2. Method for producing beverages according to the present technology
The method for producing a beverage according to the present technology is not particularly limited and can be carried out using a known method for producing a beverage. The method for producing a beverage according to the present technology preferably includes a step of mixing a sweetener, a milk protein hydrolysate, a sour component, and a flavoring to prepare a raw material liquid.

In the beverage manufacturing method of the present technology, a description of the configuration common to the above-mentioned <1. Beverage of the present technology> will be omitted.
In the manufacturing method of the beverage of the present technology, the raw materials and the amounts used thereof can be the (A) sweetener, the (B) milk protein hydrolysate, the (C) sour component, the (D) flavoring, and optional components of <1. Beverage of the present technology> described above, and the amounts used can be appropriately adjusted to achieve the respective contents and mass ratios.

In the beverage manufacturing method of the present technology, the components used in the beverage of the present technology may be added at any step in the beverage manufacturing process, and are not particularly limited, but may be added during the preparation process, for example. Furthermore, the beverage can be prepared by adding the components appropriately so that the contents and mass ratios described above are achieved.

The manufacturing process for the beverage of the present technology includes, for example, a step of preparing a raw material liquid by mixing ingredients such as the components (A) to (D) above, a step of heat sterilizing or sterilizing (membrane treatment, etc.) the raw material liquid, and a step of filling the sterilized or sterilized raw material liquid into a container. Alternatively, a heat sterilization step may be carried out after filling the raw material liquid before sterilization into a container. The manufacturing process of the present technology is not limited to these.

In the present technology, the step of adding the milk protein hydrolysate is not particularly limited as long as it occurs during the beverage production process, and may occur at any step. For example, it may be added during the preparation process, before the heat sterilization process, or before the container filling process. Furthermore, it is preferable that the heat sterilization process be carried out after the milk protein hydrolysate addition step.

Furthermore, when preparing an acidic beverage, a liquid containing an acidic component or the like may be appropriately mixed in at any step to adjust the beverage to an acidic state.

In this technology, homogenization is preferred. This homogenization can be carried out by conventional methods. For example, a method using a homogenizer at 60 to 90°C and a pressure of 5 to 25 MPa can be exemplified, but is not limited to this.

In this technology, heat sterilization or sterilization is preferred. This can be done by conventional heat sterilization or sterilization methods. In the case of heat sterilization, it is usually done at 120-150°C for 1-120 seconds, and from the perspective of beverage flavor, it is more preferable to do so at 120-140°C for 1-3 seconds. UHT sterilization (Ultra-High Temperature pasteurization) may also be used. Sterilization may also be done using a filter, etc.

In the present technology, filling into a container can be done using conventional methods. Composition containers used for beverages in the present technology include, but are not limited to, paper cartons, PET containers, cans, bottles, etc., and an appropriate container may be selected depending on the state of the beverage composition (e.g., beverage, liquid food, etc.).

The above production method can provide a beverage with an enhanced fruit juice flavor and/or an improved sweet aftertaste, and can also improve the palatability of the beverage.
The form of the beverage of the present invention is not particularly limited, and may be either liquid or fluid.
Furthermore, while heat treatment generally tends to weaken the fruit juice flavor, the present technology can enhance the fruit juice flavor and/or improve the sweet aftertaste even in beverages that have been heat-sterilized, thereby providing excellent heat-sterilized beverages.

3. Uses of this technology
The present technology can also provide a method for enhancing the fruit juice flavor of a beverage and/or a method for improving the aftertaste of a sweetener in a beverage, which method comprises adding a sweetener, a milk protein hydrolysate, a sour component, and a flavoring to the beverage.
Furthermore, the milk protein hydrolysate of the present technology can enhance the fruit juice flavor of beverages that are expected to have a fruit juice flavor (more preferably low-fruit juice or no-fruit juice beverages), and can also improve the aftertaste of sweeteners (more preferably high-intensity sweeteners).It is preferable to use the milk protein hydrolysate of the present technology in soft drinks or milk-containing beverages.

Furthermore, when using the milk protein hydrolysate of the present technology to enhance the fruit juice flavor of a beverage, it is preferable to include a sour component and a flavoring in the beverage. Furthermore, when improving the aftertaste of a sweetener, it is preferable to include a sweetener in the beverage. It is even more preferable to include three components in the beverage: a sweetener, a sour component, and a flavoring. This will further enhance the fruit juice flavor and/or improve the aftertaste of the sweetener, and also further improve the deliciousness of the beverage.

Therefore, the milk protein hydrolysate of the present technology can be contained as an active ingredient in an agent for enhancing the fruit juice flavor of beverages and/or an agent for improving the aftertaste of sweeteners, and can also be used to produce an agent for enhancing the fruit juice flavor of beverages and/or an agent for improving the aftertaste of sweeteners. The present technology can also provide a milk protein hydrolysate or use thereof for enhancing the fruit juice flavor of beverages and/or improving the aftertaste of sweeteners. The present technology can also provide a method for enhancing the fruit juice flavor of beverages and/or a method for improving the aftertaste of sweeteners by incorporating a milk protein hydrolysate into a beverage.

In the present technology, explanations of components common to the above-mentioned <1. Beverage of the Present Technology> and <2. Method for Producing a Beverage of the Present Technology> will be omitted. The amounts and ratios of the sweetener, milk protein hydrolysate, acidity component, flavoring, etc. used in the present technology are the same as the contents and mass content ratios of each component in the above-mentioned <1. Beverage of the Present Technology> and <2. Method for Producing a Beverage of the Present Technology>.

The present technology will be explained in more detail below based on examples. Note that the examples described below are merely representative examples of the present technology, and should not be construed as narrowing the scope of the present technology.

Example 1 and Comparative Example 1: Juice-free soft drink (grapefruit juice-free)
We produced juice-free soft drinks by adjusting the amount of casein hydrolysate added, and investigated the enhancement of fruit juice flavor, improvement of sweetener aftertaste, and improvement of palatability for each soft drink.

<Raw materials used in Example 1 and Comparative Example 1>
The raw materials used in Example 1 and Comparative Example 1 are as follows.
As the milk protein hydrolysate, casein protein hydrolysate (Production Example 1): the casein protein hydrolysate obtained in the following [Production Example 1] was used.
Fibersol 2 (dietary fiber; manufactured by Matsutani Chemical Industry Co., Ltd.) is a resistant dextrin.
Citric acid was used as the sour component.
The high-intensity sweetener used was Sunsweet SA5050 (manufactured by San-ei Gen FFI Co., Ltd.) Sunsweet SA5050 is a sweetener containing acesulfame potassium and sucralose, and has a sweetness intensity approximately 200 times that of sugar.
As the grapefruit flavor, a grapefruit flavor containing naturally derived ingredients manufactured by Takasago International Corporation was used.

The content of casein protein hydrolysate in the beverage was 0% by mass in Test Example 1, 0.05% by mass in Test Example 2, 0.075% by mass in Test Example 3, 0.1% by mass in Test Example 4, and 0.2% by mass in Test Example 5.
The sweetness intensity of the beverage of Test Example 1 was 6. The sweetness intensity of the beverages of Test Examples 2 to 5 was determined using this casein protein hydrolysate-free beverage.
The acidity of each of the beverages of Test Examples 1 to 5 was measured according to the above-mentioned "Japanese Agricultural Standards for Fruit Drinks" and was found to be 0.3%.

In Tables 2 and 4, "hydrolysate (%)" refers to "casein hydrolysate (mass %)." "Amount of hydrolysate per sweetness level of 1" refers to "content (mass %) of casein hydrolysate in a beverage per sweetness level of 1." "Amount of hydrolysate per [sweetness/acidity] ratio of 1" refers to "content (mass %) of casein hydrolysate in a beverage per [sweetness/acidity] ratio of 1." "Mass ratio of [acid component/hydrolysate]" refers to "mass ratio of [content (mass %) of acid component/content (mass %) of hydrolysate in a beverage." "Mass ratio of [mass %) of flavoring/hydrolysate in a beverage" refers to "mass ratio of [content (mass %) of flavoring/content (mass %) of casein hydrolysate in a beverage."

[Production Example 1: Casein Protein Hydrolysate]
(A) Casein protein hydrolysate was used as the milk protein hydrolysate.
900 mg of water was added to 100 mg of commercially available casein protein (derived from cow's milk, manufactured by New Zealand Dairy Board) and the mixture was thoroughly dispersed. Sodium hydroxide was added to adjust the pH of the solution to 7.0, and the casein protein was completely dissolved, thereby preparing an aqueous casein protein solution with a concentration of approximately 10%.
The casein protein aqueous solution was sterilized by heating at 85°C for 10 minutes, adjusted to 50°C, and sodium hydroxide was added to adjust the pH to 9.0, after which 2 mg of pancreatin (Amano Enzyme Inc.) and 4 mg of protease A (Amano Enzyme Inc.) were added to initiate the hydrolysis reaction. After 8 hours, the enzyme was inactivated by heating at 80°C for 6 minutes, and the enzymatic reaction was stopped, and the mixture was cooled to 10°C.
This hydrolyzate was ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1000 (manufactured by Nippon Pall Co., Ltd.), concentrated and then freeze-dried to obtain 85 mg of casein protein hydrolysate.

(A) The above process was carried out on multiple lots of casein protein hydrolysate, and the results were a hydrolysis rate of 20-30%, an average molecular weight of 800 Da or less, an amino acid release rate of 10% or less, and a tripeptide MKP content of 0.01-0.1% by mass. These were calculated using the above-mentioned <Amino Acid Hydrolysis Rate>, <Method for Calculating Average Molecular Weight>, <Method for Calculating Amino Acid Release Rate>, and <Measurement of Tripeptide MKP Content>. Casein protein hydrolysate can be prepared to have an average molecular weight of 360-390 Da.

<Preparation and Evaluation of Beverages of Example 1 and Comparative Example 1>
Using the above-mentioned <Raw materials used in Example 1 and Comparative Example 1> and the blending compositions of Test Examples 1 to 5 shown in Tables 1 and 2, five types of fruit juice-free soft drinks (Test Examples 1 to 5) were produced.
For the non-fruit juice soft drink of Test Example 1, a reference non-fruit juice soft drink 1 (Comparative Example 1) was prepared in the same manner as in Test Example 2 below, except that the casein protein hydrolysate of Production Example 1 was not blended in the composition of Table 1.
In the formulation of Test Example 2, a raw material liquid was prepared by mixing the high-intensity sweetener, the casein protein hydrolysate of Production Example 1, the acidic component, the flavor, water, and other raw materials, and a mixture (pH 2.5 to 4.0 (20°C)) was prepared so as to have the composition shown in the above-mentioned <Raw Materials Used in Example 1 and Comparative Example 1>, Tables 1 and 2. This prepared mixture was homogenized at 60°C and 20 MPa, further heat sterilized (UHT sterilization at 120 to 140°C for about 1 to 3 seconds), and then cooled to room temperature to obtain fruit juice-free soft drink 2 (pH 2.5 to 4.0 (20°C)).
According to the blending compositions shown in Tables 1 and 2 and the above-mentioned <Raw materials used in Example 1 and Comparative Example 1>, each blending composition of Test Examples 3 to 5 was used to obtain each of the fruit juice-free soft drinks 3 to 5.

<Evaluation method>
In order to standardize the fruit juice sensation among the evaluation panelists, natural fruit juice was used as the standard liquid, which was checked on a daily basis. Seven members of the development team who are responsible for fine-tuning the criteria for juicy sensation were selected as the evaluation panelists. This time, natural grapefruit juice was also used to fine-tune the criteria among the evaluation panelists.
Seven members of the development team, who routinely check each sample liquid with each sweetness level and also refine the criteria for judging the aftertaste of sweeteners, were selected as evaluation panelists.
In order to standardize the palatability among the evaluation panelists, we interviewed them about palatability, and selected seven of the development staff who were able to agree on the criteria for judging palatability as evaluation panelists.

<Evaluation points>
<Evaluation of fruitiness (7-point scale: 1 being the worst / 7 being the best)>
1 (very weak); 2 (weak); 3 (slightly weak); 4 (neither strong nor weak); 5 (slightly strong); 6 (strong); 7 (very strong)

<Sweetener aftertaste (stickiness) rating (7-point scale: 1 being best/7 being worst)>
1 (very weak); 2 (weak); 3 (slightly weak); 4 (neither strong nor weak); 5 (slightly strong); 6 (strong); 7 (very strong)

<Taste rating (7-point scale: 1 being the worst / 7 being the best)>
1 (very bad); 2 (bad); 3 (slightly bad); 4 (neither good nor bad); 5 (slightly good); 6 (good); 7 (very good)

<Results for the beverage of Example 1>
By adding casein hydrolysate to a soft drink containing a sweetener, a sour component, and a citrus flavor, the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the beverage of Example 1, which had improved taste, was obtained. The beverage of Example 1 was a juice-free beverage that did not contain fruit juice, but the fruit juice flavor was imparted to it, making it a beverage with a fruit juice flavor. The beverage of Example 1 was a beverage in which the unpleasant aftertaste typical of high-intensity sweeteners was improved.

In terms of enhancing the fruit juice flavor, when the casein hydrolysate content was 0.05 to 0.2% by mass, the fruit juice flavor was enhanced, and among these, when the content was 0.05 to 0.075% by mass, the enhancement of the fruit juice flavor was even better.
In improving the aftertaste of sweeteners, when the content of casein hydrolysate was 0.05 to 0.2% by mass, the aftertaste was improved, and among these, when the content was 0.1 to 0.2% by mass, the aftertaste was improved more effectively.
When the content of casein hydrolysate was 0.05 to 0.05% by mass, the improvement in palatability was more excellent.
Considering the overall situation, it was found that the casein hydrolysate content was preferably 0.05 to 0.075% by mass.

When the content of casein hydrolysate in the beverage relative to a sweetness level of 1 was 0.008 to 0.016 mass % relative to a sweetness level of 1, the enhancement of the fruit juice flavor was more satisfactory.
When the content of casein hydrolysate in the beverage relative to a sweetness/acidity ratio of 1 was 0.0025 to 0.00375 mass%, the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the taste was improved.
When the mass ratio of the sour component content to the casein hydrolysate content was 6.00 to 1.5 (more preferably 6.00 to 4.00), the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the taste was improved.
When the mass ratio of flavor content to casein hydrolysate content was 1.20 to 0.30 (more preferably 1.20 to 0.80), the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the taste was improved.

Example 2 and Comparative Example 2: Fruit Juice-Free Milk-Containing Beverage (Grapefruit-Flavored Milk-Containing Beverage)
We produced non-fruit juice milk-containing beverages by adjusting the amount of casein hydrolysate added, and examined each milk-containing beverage to enhance the fruit juice flavor, improve the aftertaste of the sweetener, and improve the palatability.

<Ingredients used in the beverages of Example 2 and Comparative Example 2>
The raw materials used in Example 2 and Comparative Example 2 are as follows.
As the milk protein hydrolysate, casein protein hydrolysate (Production Example 1): the casein protein hydrolysate obtained in the above [Production Example 1] was used.
As sweeteners, sugars such as high fructose corn syrup (manufactured by Showa Sangyo Co., Ltd.) and granulated sugar (manufactured by Mitsui Sugar Co., Ltd.) were used.
As milk components, skim milk powder (manufactured by Morinaga Milk Industry Co., Ltd.) and whey powder (manufactured by Morinaga Milk Industry Co., Ltd.) were used.
Citric acid and trisodium citrate were used as sour components.
As the grapefruit flavor, a flavor (manufactured by Takasago International Corporation) was used.
As stabilizers, soybean polysaccharides and pectin (manufactured by San-Ei Gen FFI Co., Ltd.) were used.
As a coloring agent, carminic acid pigment manufactured by San-Ei Gen FFI Co., Ltd. was used.

The sweetness intensity of the beverage of Test Example 6 was 11. The sweetness intensity of the beverages of Test Examples 7 to 11 was determined using this casein protein hydrolysate-free beverage.
As a result of measurement according to the above-mentioned "Japanese Agricultural Standards for Fruit Drinks," the acidity (%) of the drinks of Comparative Example 2 and Example 2 (Test Examples 6 to 11) was 0.22.
The content of casein protein hydrolysate in the beverage was 0% by mass in Test Example 6, 0.1% by mass in Test Example 7, 0.2% by mass in Test Example 8, 0.3% by mass in Test Example 9, 0.4% by mass in Test Example 10, and 0.5% by mass in Test Example 11.
The beverages of Example 2 (Test Examples 6 to 11) were measured according to the "Test Method for Compositional Standards of Milk, etc." in the "Ministerial Ordinance on Milk, etc. (Ministerial Ordinance on Compositional Standards, etc. of Milk and Dairy Products)" mentioned above. As a result, the non-fat milk solids content was 0.54% by mass, and the milk fat content was 0.01% by mass.

<Preparation and Evaluation of Beverages of Example 2 and Comparative Example 2>
Six types of milk-containing beverages (Test Examples 6 to 11) were produced using the above-mentioned <Raw materials used in Example 2 and Comparative Example 2> and the blending compositions of Test Examples 6 to 11 shown in Tables 3 and 4.
For the fruit juice-free milk-containing beverage of Test Example 6, a reference fruit juice-free milk-containing beverage 6 (Comparative Example 2) was prepared in the same manner as in Test Example 7 below, except that the casein protein hydrolysate of Production Example 1 was not blended in the composition of Table 2.
In the formulation of Test Example 7, a raw material liquid was prepared by mixing sugars, milk components, the casein protein hydrolysate of Production Example 1, a sour component, flavorings, water, and other raw materials, and a mixture (pH 2.5 to 4.0 (20°C)) was prepared so as to have the composition shown in the above-mentioned <Raw materials used in Example 2 and Comparative Example 2>, Tables 3 and 4. This prepared mixture was homogenized at 60°C and 20 MPa, further heat sterilized (UHT sterilization at 120 to 140°C for about 1 to 3 seconds), and then cooled to room temperature to obtain milk-containing beverage 7 (pH 2.5 to 4.0 (20°C)).
According to the blending compositions shown in Tables 3 and 4 of the above-mentioned <Raw materials used in Example 2 and Comparative Example 2>, milk-containing beverages 8 to 11 were obtained in turn using the blending compositions of Test Examples 8 to 11.

<Evaluation method and evaluation points for the beverages of Example 2 and Comparative Example 2>
The beverages of Example 2 and Comparative Example 2 were evaluated using the same methods and with the same evaluation points as those described above in <Preparation and Evaluation of Beverages of Example 1 and Comparative Example 1>.

<Results for the beverage of Example 2>
By blending casein hydrolysate into a milk-containing beverage containing a sweetener, a sour component, and a citrus flavor, the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the beverage of Example 2 was obtained, which had improved taste. The beverage of Example 2 was a non-fruit juice beverage that did not contain fruit juice, but was imparted with a fruit juice flavor, making it a beverage with a fruit juice flavor. The beverage of Example 2 was a milk-containing beverage, but was able to be made into a good milk-containing beverage with a fruit juice flavor even though it contained milk components.

In terms of enhancing the fruit juice feel, when the casein hydrolysate content was 0.1 to 0.5% by mass, the fruit juice feel was enhanced, and within this, when the content was 0.2 to 0.5% by mass (preferably 0.2 to 0.4% by mass), the enhancement of the fruit juice feel was even better.
In improving the aftertaste of sweeteners, when the content of casein hydrolysate was 0.2 to 0.5% by mass, the aftertaste of sweeteners was improved, and among these, when the content was 0.3 to 0.5% by mass, the improvement in the aftertaste of sweeteners was more satisfactory.
When the content of casein hydrolysate was 0.1 to 0.5% by mass, the improvement in palatability was more excellent.
Considering all factors, the content of casein hydrolysate was preferably 0.3 to 0.5% by mass, more preferably 0.3 to 0.4% by mass.

When the content of casein hydrolysate in the beverage relative to a sweetness level of 1 was 0.009 to 0.045 mass %, the enhancement of the fruit juice flavor was more satisfactory.
When the content of casein hydrolysate in the beverage relative to a sweetness/acidity ratio of 1 was 0.002 to 0.01 mass %, the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the taste was improved.
When the mass ratio of the sour component content to the casein hydrolysate content was 2.90 to 0.58 (more preferably 0.97 to 0.58), the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the palatability was improved.
When the mass ratio of flavor content to casein hydrolysate content was 1.10 to 0.22 (more preferably 0.37 to 0.28), the fruit juice flavor was enhanced, the aftertaste of the sweetener was improved, and the taste was improved.

<Prescription example>
Examples of production and formulation of the food and beverage composition of the present technology are shown below, but the composition of the present technology is not limited to these.

<Formulation Examples 1 and 2: Fruit Juice-Free Soft Drink/Milk-Containing Beverage>
A fruit juice-free soft drink of Formulation Example 1 can be obtained in the same manner as Test Example 2, except that lemon flavoring is used instead of the grapefruit flavoring of Test Example 2. The soft drink of Formulation Example 1 exhibits an enhanced fruit juice flavor, an improved aftertaste of the sweetener, and improved palatability.
A fruit juice-free milk-containing beverage of Formulation Example 2 can be obtained in the same manner as Test Example 8, except that lemon flavoring is used instead of the grapefruit flavoring of Test Example 8. The milk-containing beverage of Formulation Example 2 exhibits an enhanced fruit juice flavor, an improved aftertaste of the sweetener, and improved palatability.

<Formulation Example 3: Low-fruit juice soft drink>
A low-fruit juice soft drink of Formulation Example 3 can be obtained by substituting commercially available 100% concentrated grapefruit juice (with flavoring) in place of the grapefruit flavoring of Test Example 3, adjusting the content to be within the range of 5% to less than 10%. Formulation Example 3 uses the casein protein hydrolysate (Production Example 1) and its amount (0.075%), as in Test Example 3. This production method can be carried out in accordance with the production method of Test Example 3. This low-fruit juice soft drink can be obtained by adjusting the sweetness level to 6, the amount of hydrolysis relative to sweetness level 1 to 0.012%, and the [sweetness level/acidity level] to 20.
This low-fruit juice soft drink has an enhanced fruit juice flavor, an improved sweetener aftertaste, and improved palatability.

Claims (11)

A beverage comprising a sweetener, a milk protein hydrolysate, an acidic component and a flavoring,
The mass ratio of [the content of the sour component/the content of the milk protein hydrolysate] is 6 to 0.5,
The beverage has a fruit juice flavor and an alcohol content of less than 1%, and the content of the milk protein hydrolysate is 2% by mass or less. The beverage according to claim 1, wherein the milk protein hydrolysate is casein protein hydrolysate. The beverage according to claim 1 or 2, wherein the milk protein hydrolysate is a milk protein hydrolysate containing Met-Lys-Pro. The beverage according to any one of claims 1 to 3, wherein the milk protein hydrolysate content is 0.05 to 0.5% by mass. The beverage according to claim 3 , wherein the content of Met-Lys-Pro in the beverage is 0.00002 to 0.001% by mass. The beverage according to claims 1 to 5, wherein the milk protein hydrolysate content relative to a sweetness level of 1 of the beverage is 0.004 to 0.1% by mass. The beverage according to any one of claims 1 to 6, wherein the flavoring is a citrus flavoring. The beverage described in any one of claims 1 to 7, wherein the beverage is a soft drink or a milk-containing beverage. The beverage described in any one of claims 1 to 8, wherein the beverage is a low-fruit juice beverage or a fruit juice-free beverage. The method includes a step of preparing a raw material liquid by mixing a sweetener, a milk protein hydrolysate, a sour component, and a flavoring,
In the step of preparing the raw material liquid, the mass ratio of [content of sour component/content of milk protein hydrolysate] is 6 to 0.5,
The sour component and the milk protein hydrolysate are mixed so that the content of the milk protein hydrolysate is 2% by mass or less.
A method for producing a beverage having an alcohol content of less than 1% and a fruit juice flavor . The present invention is characterized in that a sweetener, a milk protein hydrolysate, an acidic component and a flavoring are added to a beverage having an alcohol content of less than 1% and a fruit juice flavor ,
The mass ratio of [content of sour component/content of milk protein hydrolysate] is 6 to 0.5,
A method for enhancing the fruit juice flavor of a beverage, comprising adding the sour component and the milk protein hydrolysate so that the content of the milk protein hydrolysate is 2% by mass or less.