JP2020150936A - Momordica grosvenori extract - Google Patents

Abstract

To provide a Momordica grosvenori extract with reduced water-insoluble matter.SOLUTION: A Momordica grosvenori extract is prepared that contains (1) mogroside V of 0.01 mass%-70 mass% and (2) a water-insoluble component of 5 mass% or less, with the content of water-insoluble component/the content of polyphenol of 1/2 or less. Such a Momordica grosvenori extract can be used as a food and drink composition, a pharmaceutical composition, a cosmetic composition or an oral care preparation.SELECTED DRAWING: Figure 1

Description

The present invention relates to Luo Han Guo extract. More specifically, the present invention relates to a Luo Han Guo extract in which the amount of water-insoluble components is suppressed.

Luo Han Guo (scientific name: Siraita Grosvenorii (Swingle) C. Jeffrey ex AM Lu & Zhi Y. Zhang (Momordica grossvenorii Swingle) is a perennial plant native to the Guangxi Zhuang Autonomous Region in southern China. Luo Han Guo has a large temperature difference between day and night, high humidity, high fog, fertile soil, rich in corrosion, sufficient water is obtained, drainage is excellent, breathability is high, and the outbreak of pests is low. It grows only in a limited area (Non-Patent Document 1).

Luo Han Guo is heated and dried, and is used as a luxury item to enjoy the sweetness and aroma like brown sugar. The decoction of dried fruits is also known to have effects such as coughing, heat cooling, and relief of inflammation. Furthermore, it can be used during pregnancy and when physical strength is weakened after childbirth, and it is excellent in nourishing tonicity. (Non-Patent Document 2). In recent years, it has also been elucidated that it has an action of promoting wound healing (Patent Document 1).

From the fruits of Luo Han Guo, it is possible to obtain an extract of Luo Han Guo, which has an extremely strong sweetness and a unique flavor with a deep richness. The main components of the sweetness of this Luo Han Guo extract are fructose and triterpene glycosides called mogrosides (Non-Patent Document 3). Although mogrosides are non-carbohydrate-based, they have a sweetness, and the sweetness is mainly derived from a compound called mogroside V. In addition to mogrosides, Luo Han Guo extract contains components such as sugars, sugars, proteins, amino acids, organic acids, dietary fiber, and minerals. The color is yellow to brown, and the liquid is opaque. It's cloudy. It is known that mogroside V is 300 times or more sweeter than sucrose and exhibits excellent sweetness among high-sweetness sweeteners (Non-Patent Document 4).

JP-A-2009-0573336

Luo Han Guo Cultivation Chemistry Research (2010) Illustrated Chinese Medicine Dictionary Volume 4 (1982) Journal of Pharmacy-103 (11) -1151-1154 (1983) Japan Food Science and Engineering Magazine 53-10 (2006)

While Luo Han Guo extract is thus very useful, it is very difficult to obtain a satisfactory product that has all of the physical properties, taste, and price. In particular, it is possible to increase the amount of mogroside V, which is the main sweetening component in Luo Han Guo extract, by repeating the purification process, but on the other hand, it is useful for proteins, minerals, etc. at multiple purification stages. Ingredients may be lost or the taste may be out of balance.

An object of the present invention is to provide a Luo Han Guo extract having excellent physical properties and taste.

As a result of diligent studies to solve the above problems, the present inventors retained mogroside V, which is the main sweetening component, and useful components by suppressing the amount of water-insoluble components that may occur in the Luo Han Guo extract. However, they have found that a Luo Han Guo extract having excellent properties such as a clear liquid property, which has been found only in a highly purified Luo Han Guo extract, can be obtained, and have completed the present invention.

That is, the present invention provides the following Luo Han Guo extracts.
Item 1.
Luo Han Guo (Siraita grosvenorii) extract,
(1) Mogroside V is contained in an amount of 0.01% by mass to 70% by mass; and (2) a water-insoluble component is contained in an amount of 5% by mass or less.
Further, a Luo Han Guo extract having an insoluble polyphenol content / total polyphenol content of 1/2 or less.
Item 2.
Item 2. The Luo Han Guo extract according to Item 1, wherein the absorbance at a wavelength of 660 nm in a 10 mass% aqueous solution is 0.3 or less.
Item 3.
Item 2. The Luo Han Guo extract according to any one of Items 1 or 2, wherein the total polyphenol content is 0.1% by mass to 20% by mass.
Item 4.
Item 2. The Luo Han Guo extract according to any one of Items 1 to 3, wherein the insoluble polyphenol content is 5% by mass or less.
Item 5.
Item 6. The Luo Han Guo extract according to any one of Items 1 to 4, wherein the total protein content is 5% by mass to 90% by mass.
Item 6.
Item 6. The Luo Han Guo extract according to any one of Items 1 to 5, wherein the insoluble protein content is 3% by mass or less.
Item 7
A food or drink composition containing the Luo Han Guo extract according to any one of Items 1 to 6.
Item 8.
A pharmaceutical composition containing the Luo Han Guo extract according to any one of Items 1 to 6.
Item 9.
A cosmetic composition or an oral care preparation containing the Luo Han Guo extract according to any one of Items 1 to 6.
Item 10.
It is an extract of Luo Han Guo
A process of obtaining a crude extract by treating Luo Han Guo with a solvent;
Luo Han Guo extract obtained by a method including a step of cooling the extract to 40 ° C. or lower within 120 minutes from the start of solvent treatment; and a step of solid-liquid separation at 40 ° C. or lower.
Item 11.
(1) A step of obtaining a crude extract by treating Luo Han Guo with a solvent;
(2) A method for producing a Luo Han Guo extract, which comprises a step of adjusting the crude extract to 40 ° C. or lower within 120 minutes from the start of solvent treatment; and (3) a step of centrifuging at a temperature of 40 ° C. or lower.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a Luo Han Guo extract in which the amount of water-insoluble components is suppressed.

FIG. 1 is a diagram showing a state of an aqueous solution containing 1% by mass of the dry mass of each of the Luo Han Guo extracts of Examples and Comparative Examples. FIG. 2 is a diagram schematically showing and comparing the production times of Luo Han Guo extracts of Examples and Comparative Examples.

In the present specification, the unit of content "mass%" is synonymous with "g / 100g".

The Luo Han Guo extract of the present invention
Luo Han Guo (Siraita grosvenorii) extract,
(1) Mogroside V is contained in an amount of 0.01% by mass to 70% by mass; and (2) a water-insoluble component is contained in an amount of 5% by mass or less.
Further, the insoluble polyphenol content / total polyphenol content is 1/2 or less.

[Luo Han Guo extract]
Luo Han Guo (Monk Fruit) is a plant of the Cucurbitaceae family, and its scientific name is Siraita grosvenorii. It is characterized by containing a sweet component called triterpene glycoside. When Luo Han Guo became widely recognized as a commercial crop, demand increased rapidly, and along with this, the cultivation location changed significantly from mountainous areas to flat areas from the viewpoint of efficiency. Luo Han Guo is basically a crop that can be harvested once a year. Seedlings can be planted in April and harvested in October. There are various varieties in cultivated areas such as Asia and North America, but in the present invention, any varieties in any cultivated area can be used.

"Luo Han Guo extract (Monk Fruit Extract)" is a general term for extracts of Luo Han Guo, and has forms such as liquid, semi-solid, and solid. It is preferably a solid such as powder. It refers to those in which the components other than the solvent are 100% derived from Luo Han Guo. Examples of the solvent for extraction include water, methanol, ethanol, carbon dioxide and the like, and are not particularly limited. In addition, as a position in the narrow sense of "Luo Han Guo extract", there is "Luo Han Guo Extract", which is a Luo Han Guo extract that meets the standards described in the official standard for food additives in Japan. The Luo Han Guo extract of the present invention includes, but is not limited to, the Luo Han Guo extract satisfying such a regulation.

((A) Mogroside V)
Examples of mogrosides contained in the Luo Han Guo extract of the present invention include mogroside V, mogroside IV, 11-oxomogroside V, and siamenocid I, and proteins are also included.

Mogroside V is a compound represented by the following chemical formula, and is a substance exhibiting particularly excellent sweetness among the components contained in Luo Han Guo extract.

Due to these properties, it may be used as a component of high-intensity sweeteners.

In the Luo Han Guo extract of the present invention, the content of the component (A) with respect to the total amount of the extract can be determined by the extraction method from Luo Han Guo, but can be appropriately set by the purification method. The content of the component (A) is 0.01% by mass to 70% by mass, preferably 0.5% by mass to 66% by mass, and more preferably 0.1% by mass, based on the total amount of the composition. It is from mass% to 60% by mass.

In the Luo Han Guo extract of the present invention, the content of mogrosides with respect to the total amount of the extract can be determined by the extraction method from Luo Han Guo, but can be appropriately set by the purification method. The content of mogrosides is 0.01% by mass to 88% by mass, preferably 0.5% by mass to 83% by mass, and more preferably 0.1 to 75% by mass, based on the total amount of the composition. It is mass%.

(Mogroside V measurement method)
The amount of mogroside V contained in the Luo Han Guo extract of the present invention can be measured by using high performance liquid chromatography under the conditions shown below.

((B) Water-insoluble component)
The water-insoluble component contained in the Luo Han Guo extract of the present invention is a value that can be obtained as follows. The Luo Han Guo extract is dried in advance at 105 ° C. to a constant weight, the dry weight is determined, and 1 g of the dry weight is weighed (weighed mass value T). This is placed in a 15 mL centrifuge tube (BioLite 15 mL Clinical Centrifuge Tubes; manufactured by Thermo Fisher Scientific Co., Ltd.) (empty mass value b), and 9 g of distilled water (water temperature 20 to 25 ° C.) is added. In a vortex mixer, suspend the powder until it becomes uniform throughout the aqueous solution, allow it to stand, and then centrifuge at 4,000 rpm for 5 minutes (desktop centrifuge H-36α; manufactured by Kokusan Co., Ltd.) to force insolubilization. Settle things. Then, only the supernatant is removed, and 9 g of distilled water is newly added and suspended by pipetting. Centrifuge is performed in the same manner, the supernatant is removed, and only the precipitate obtained at this time is frozen at −20 ° C. for 12 to 24 hours (freezer SCR-SK231; manufactured by Sanyo Co., Ltd.) and then freeze-dried (frozen). After vacuum dryer VD-800R; manufactured by TIETECH Co., Ltd.), weigh (mass value c for each centrifuge tube after freeze-drying). For mass measurement in this test, record up to 4 digits after the decimal point using a precision balance (ML104T / 00; manufactured by METTLER TOLEDO). Calculate the proportion of insoluble matter using the calculation formula shown in Equation 1 for each recorded mass value. The same operation is performed three times, and the average value is calculated to obtain the ratio of insoluble components.
Percentage of insoluble components (%) = (cc) / T × 100 (Equation 1)

In the Luo Han Guo extract of the present invention, the total content of the (B) water-insoluble component with respect to the total amount of the composition is 5% by mass or less, preferably 3% by mass or less, from the viewpoint of the effect of improving physical properties and taste. , More preferably 2% by mass or less, still more preferably 1% by mass or less, even more preferably 0.5% by mass or less, particularly preferably zero or an undetectable amount. However, the lower limit may be about 0.01% by mass.

In the Luo Han Guo extract of the present invention, the ratio of the content of the water-insoluble component to the mogroside V is not particularly limited. Usually, the content of mogroside V in a certain composition increases according to the degree of purification of Luo Han Guo extract, and a composition having a low content of mogroside V means that the degree of purification is low and is water-insoluble. It tends to contain a lot of ingredients. Therefore, in the compositions containing the Luo Han Guo extract so far, the water-insoluble component tends to decrease as the amount of mogroside V increases. However, in the present invention, the amount of the water-insoluble component is suppressed to a certain level or less even when the water-insoluble component is about 0.01% by mass to 20% by mass of the whole composition.

(Insoluble polyphenol content / total polyphenol content)
The ratio of insoluble polyphenol content / total polyphenol content in the Luo Han Guo extract of the present invention is the ratio of the amount of polyphenol contained in the entire Luo Han Guo extract to the amount of polyphenol contained in the insoluble component in the same amount of Luo Han Guo extract. Calculate by finding. The amount of each polyphenol is a value that can be obtained as follows. That is, FOLIN-DENIS REAGENT is prepared. To 188.0 mL of distilled water, 27.73 g of sodium tungstate dihydrate, 5 g of phosphomolybdic acid and 12.5 mL of 85% phosphoric acid are added. The mixed solution is refluxed in a fume hood using a Liebig condenser for 2 hours. After the reflux operation, allow to cool, and after the liquid temperature reaches room temperature, transfer to a 1000 mL volumetric flask and fill up with distilled water up to the marked line. This solution (FOLIN-DENIS reagent) is subdivided into a light-shielding container and stored in a cool and dark place until use. Separately, weigh 0.1 g of Luo Han Guo extract and dissolve in 1 mL of water. To a 2 mL microtube, add 0.5 mL of FOLIM-DENIS REAGENT and 0.5 mL of diluted Luo Han Guo extract, and after 3 minutes, add a 10 w / v% sodium carbonate aqueous solution. The mixture is centrifuged after 60 minutes (14,000 G, 5 minutes, room temperature). The absorbance at 760 nm is measured with respect to the supernatant of the sample after centrifugation. Distilled water is used as the blank solution, and each Luo Han Guo extract is adjusted and measured so that the absorbance is in the range of 0.100 to 1.000 according to Lambert-Beer's law. Similarly, a calibration curve (R2 ≧ 0.999) is prepared using tannic acid as a standard substance, a 0.1 mg / mL tannic acid aqueous solution is prepared, and the mixture is appropriately diluted. In this way, the total polyphenol content and the insoluble polyphenol content are determined using the polyphenol concentration in the sample obtained from the calibration curve. The same operation is performed three times to obtain the average value, and then the content ratio between the average values is defined as the insoluble polyphenol content / total polyphenol content. The total polyphenol content is a value obtained by measuring the amount of polyphenol using Luo Han Guo extract as a sample, and the insoluble polyphenol content is a value obtained by measuring the polyphenol in the water-insoluble component.

In the Rakan fruit extract of the present invention, the total polyphenol content is preferably 0.1% by mass to 20% by mass, more preferably 0.3% by mass to 15% by mass, and even more preferably 0.5% by mass. % To 10% by mass, most preferably 0.7% by mass to 8% by mass.

In the Rakan fruit extract of the present invention, the insoluble polyphenol content is preferably 0.001% by mass to 5% by mass, more preferably 0.002% by mass to 2% by mass, and even more preferably 0.00. It is 3% by mass to 15% by mass, most preferably 0.004% by mass to 1.2% by mass.

In the Luo Han Guo extract of the present invention, the insoluble polyphenol content / total polyphenol content is 1/2 or less, that is, 0.5 or less. The insoluble polyphenol content / total polyphenol content is preferably 0.3 or less, more preferably 0.2 or less, still more preferably 0.1 or less, still more preferably 0.05 or less.

(Insoluble protein content / total protein content)
The ratio of insoluble protein content / total protein content in the Luo Han Guo extract of the present invention is a value that can be obtained as follows. That is, 0.1 g of each Rakan fruit extract is weighed, placed in a 15 mL centrifuge tube, 0.9 g of 0.9% sodium chloride aqueous solution is added, and the mixture is suspended with a vortex mixer until the powder is homogenized throughout the aqueous solution. It becomes cloudy. Weigh 0.1 g of this solution and add 9.9 g of 0.9% aqueous sodium chloride solution (total protein content).

Weigh 0.1 g of each Rakan fruit extract, place in a 15 mL centrifuge tube, add 0.9 g of 0.9% sodium chloride aqueous solution, and suspend with a vortex mixer until the powder is homogenized throughout the aqueous solution. .. After standing, centrifuge at 4,000 rpm for 5 minutes to forcibly settle the insoluble matter. Then, only the supernatant is carefully removed, and the precipitate is suspended and dissolved in 1.0 g of a 0.9% aqueous sodium chloride solution (insoluble protein).

These were performed using a BCA protein assay (Thermo Fisher Science). The Working requirement is adjusted by mixing Solution A and Solution B in a ratio of 50: 1. Mix 50 microliters of sample with 1 milliliter of Working reagent. The mixture was heated at 37 ° C. for 30 minutes and then allowed to stand until it returned to room temperature, and the absorbance at a wavelength of 562 nm was measured. At the same time, a calibration curve (R2 ≧ 0.999) is prepared using bovine serum albumin as a standard substance. The concentration of the sample is appropriately adjusted so as to be within the range of the calibration curve.

The mass in the insoluble matter is calculated using the protein concentration in the sample obtained from the calibration curve, and the content in the Luo Han Guo extract is calculated. Perform the same operation three times to obtain the average value.

In the Luo Han Guo extract of the present invention, the total protein content is preferably 5% by mass to 90% by mass, more preferably 5% by mass to 80% by mass, and even more preferably 5% by mass to 70% by mass. Most preferably, it is 5% by mass to 60% by mass.

In the Rakan fruit extract of the present invention, the insoluble protein content is preferably 0.01% by mass to 3% by mass, more preferably 0.01% by mass to 1% by mass, and even more preferably 0.01% by mass. % To 0.5% by mass, most preferably 0.01% by mass to 0.1% by mass. It is also preferable that the insoluble protein is substantially absent in the Luo Han Guo extract of the present invention.

The ratio of insoluble protein content / total protein content is not limited, but is preferably 0.002 or less, more preferably 0.001 or less.

(Aqueous solution absorbance)
The Luo Han Guo extract of the present invention is a clear liquid in which turbidity is suppressed even when adjusted to an aqueous solution. The clarity of the Luo Han Guo extract of the present invention is a value that can be obtained as follows. That is, each Luo Han Guo extract is dried at 105 ° C. to make it constant, the dry weight is determined, and 1 g of the dry weight is weighed. This is placed in a 15 mL centrifuge tube, and 9 g of distilled water (water temperature 20 to 25 ° C.) is added. Mix with a vortex mixer until the powder is homogenized throughout the aqueous solution, and this mixing operation is completed in 1 minute. Measure immediately after preparing the sample. For the measurement, 3 mL of the liquid is filled in a disposable cell (optical path length 10 mm, optical path width 10 mm), and the absorbance at 660 nm is measured using an ultraviolet-visible spectrophotometer (UV2450; manufactured by Shimadzu Corporation). Distilled water is used as the blank solution. The absorbance is measured by adjusting each Luo Han Guo extract so that it is in the range of 0.100 to 1.000 according to Lambert-Beer's law. The obtained absorbance value is corrected to the absorbance value at 10% by mass of the Luo Han Guo extract in consideration of the weighed amount of the sample using the calculation formula shown in Equation 2. Perform the same operation three times to obtain the average value.

Absorbance of sample Ab
Absorbance of blank Bab
Weighing amount (g) M

Absorbance = (Aab-Bab) x (1 / M) (Equation 2)

In the Luo Han Guo extract of the present invention, the absorbance of the 10 mass% aqueous solution having a wavelength of 660 nm, which can be obtained in this way, is preferably 0.3 or less, more preferably 0.25 or less, and further. More preferably, it is 0.21 or less, and most preferably 0.2 or less.

The Luo Han Guo extract of the present invention is also preferably a visually clear solution when adjusted to a 10% by mass aqueous solution. A clear solution is a state in which insoluble matter such as suspended matter, precipitate, and deposit derived from Luo Han Guo extract cannot be visually confirmed in the aqueous solution. It is preferable that the clarity is not affected visually by the shade of the color tone of the aqueous solution.

(Form of Luo Han Guo extract)
The form of the Luo Han Guo extract of the present invention may be liquid, semi-solid, or solid. When solid, it can be in the form of powder or granules.

[Manufacturing method of Luo Han Guo extract]
The method for producing the Luo Han Guo extract of the present invention is not particularly limited, and any method may be used as long as the liquid extracted by the solvent of Luo Han Guo is not exposed to a high temperature environment for a long period of time. Exposure to high temperature environments causes water-insoluble components to form in the Luo Han Guo extract over time. Therefore, in the method for producing Luo Han Guo extract of the present invention, first, at the stage of extraction treatment, the temperature of the extract is efficiently lowered after extraction without excessive heating, and the formation of insoluble matter is prevented. It is characterized by that.

In a preferred embodiment of the present invention, the Luo Han Guo extraction step is performed without drying the Luo Han Guo fruit after harvesting. By using the fruit of Luo Han Guo that has not been dried as a raw material, it is possible to obtain an extract of Luo Han Guo having good sweetness. The water content of the fruit of Luo Han Guo used in the present invention is typically about 30%, preferably about 50% or more, and more preferably about 70% or more. In one preferred embodiment, the water content of the fruit of Luo Han Guo is about 70% -80%. It is preferable to use the ripe Luo Han Guo fruit as a raw material. Those skilled in the art can empirically judge that Luo Han Guo is fully ripe from the color of the fruit and the shape of the fruit.

After washing and crushing the fruits of Luo Han Guo, extraction treatment is performed with water alone or with a solvent containing a lower alcohol such as ethanol or methanol. When a lower alcohol such as ethanol or methanol is used, its concentration can be about 0.01% by mass to 10% by mass. That is, it can be a mixed solvent of water and a lower alcohol. In order to improve the extraction efficiency, a method of increasing the temperature of the solvent to extract the Luo Han Guo extract is preferably used. For example, extraction using hot water is typically exemplified, but the present invention is not limited to this. Luo Han Guo fruit is divided into an extract and a Luo Han Guo residue consisting of peels and seeds.

Luo Han Guo extract can be produced by performing steps such as filtration, column absorption, column separation, recovery, concentration, and drying of the extract thus obtained. When the extraction with a solvent is hot water or a lower alcohol-containing water extraction treatment with an increased temperature, the treatment temperature and time are adjusted within a range that does not excessively heat, and after the extraction treatment, the temperature is quickly lowered to remove the water-insoluble components. Formation is suppressed.

Here, the temperature of hot water treatment or solvent treatment is not limited. For example, it is preferable that the mass ratio of the fruit of Luo Han Guo to the solvent is 1: 3 to 1:10 for about 10 to 70 minutes with water at a temperature of 75 to 100 ° C. or a solvent other than water.

“Rapidly lowering the temperature after extraction” means keeping the temperature at 40 ° C. or lower, preferably within 120 minutes, more preferably within 80 minutes, further preferably within 70 minutes, and most preferably within 65 minutes from the start of extraction. Say that.

The means for quickly lowering the liquid temperature after extraction is not particularly limited, but for example, a hot water extract of Luo Han Guo can be used in a liquid temperature control system using a liquid feeding pipe. Thereby, it is possible to quickly lower the temperature of the treated liquid to 40 ° C. or lower. That is, the liquid temperature can be maintained at 40 ° C. or lower by using a liquid feeding pipe while supplying the liquid obtained by extracting Luo Han Guo with hot water to the stock tank.

Under such conditions, steps such as fruit washing, solvent extraction, filtration and / or centrifugation, optionally concentration, column purification, spray drying, and heat sterilization may be continuously performed. In addition to the above steps, in order to further improve quality, countercurrent extraction, membrane filtration, liquid phase partitioning, two-layer separation, decolorization purification, centrifugation, recrystallization, activated carbon treatment, supercritical extraction, supercritical purification, etc. However, the content of mogloside V, which is a sweetening component of Rakanka, is adjusted to about 70% by mass at the maximum.

In the present invention, it can be a Luo Han Guo extract that has undergone steps such as fruit washing, solvent extraction, filtration and / or centrifugation, optionally concentration, column purification, spray drying, and heat sterilization.

In some cases, in addition to the above treatment, manufacturing processes such as countercurrent extraction, membrane filtration, liquid phase partitioning, two-layer separation, decolorization purification, centrifugation, recrystallization, activated carbon treatment, supercritical extraction, and supercritical purification may be combined. Is also possible.

[Sweetener composition]
The Luo Han Guo extract of the present invention can be prepared into a sweetener composition. The sweetener composition comprises the Luo Han Guo extract of the present invention, or optionally contains an auxiliary sweetening component or other components in addition to the Luo Han Guo extract of the present invention. Here, other components include, but are not limited to, excipients and diluents.

Here, the auxiliary sweetener component means a component having sweetness or adjusting the sweetness intensity and sweetness of the sweetener, and is a sugar-based sweetener such as monosaccharide, disaccharide, oligosaccharide, and sugar alcohol; Sugar-based sweeteners; and dietary fiber can be mentioned.

Among the sugar-based sweeteners, examples of monosaccharides include fructose, glucose, xylose, sorbose, galactose, and isomerized sugar. Examples of the disaccharide include maltose, lactose, trehalose, sucrose, isomerized lactose, palatinose and the like. Examples of oligosaccharides include xylooligosaccharides, fructooligosaccharides, soybean oligosaccharides, isomaltoligosaccharides, lactosucrose, galactooligosaccharides, lactulose, palatinose oligosaccharides, sucrose oligosaccharides, thean oligosaccharides, and seaweed oligosaccharides. Examples of sugar alcohols include maltitol, xylitol, sorbitol, mannitol, palatinit and the like. Among the non-carbohydrate-based sweeteners, examples of synthetic sweeteners include aspartame, advantame, acesulfame K, sucralose, and neotame. Further, among the non-carbohydrate-based sweeteners, examples of natural sweeteners include licorice extract, stevia extract, and thaumatin extract.

Dietary fibers include water-soluble dietary fibers such as pectin, guarine enzyme degradation products, glucomannan, β-glucan, polydextrose, inulin, agarose, sodium alginate, carrageenan, fucoidan, porphyran, laminarin, and indigestible dextrin; Insoluble dietary fibers such as cellulose, hemicellulose, lignin, chitin, and chitosan can be mentioned. A water-soluble dietary fiber is preferable.

When the sweetener composition of the present invention contains an auxiliary sweetening component in addition to the Rakan fruit extract, the proportion of the Rakan fruit extract contained in 100% by mass of the sweetening composition is 0.01% by mass or more. 100% by mass, preferably 0.1% by mass to 99.9% by mass, more preferably 0.1% by mass to 90% by mass, still more preferably 0.1% by mass to 50% by mass, and a particularly preferable range is 0. 1% by mass to 30% by mass can be mentioned, and the ratio of the auxiliary sweetening component is 0.01% by mass to 99.99% by mass, preferably 0.1% by mass to 99.9% by mass, and more preferably 1. Mass% to 99% by mass, more preferably 5% by mass to 99% by mass, and particularly preferably 5% by mass to 90% by mass.

As described above, the sweetener composition of the present invention comprises the Luo Han Guo extract of the present invention or contains an auxiliary sweetening component in addition to the Luo Han Guo extract, and is highly safe and has a reduced sweetness. It has no (miscellaneous taste) and has a high-quality sweetness close to that of sugar. Therefore, it can be further suitably used as a sweetener for food and drink compositions, pharmaceutical compositions (pharmaceutical products, quasi-drugs), or cosmetic compositions.

The sweetener composition of the present invention, when it consists of the Luo Han Guo extract of the present invention, has a sweetness of 200 to 400 times that of sugar (sucrose). The degree of sweetness can be appropriately adjusted by adding an auxiliary sweetness component to the Luo Han Guo extract.

According to the Luo Han Guo extract prepared by the method of the present invention, the Luo Han Guo extract of the present invention, and the sweetener composition of the present invention, a low energy (low calorie) or zero energy (zero calorie) sweetener composition can be obtained. It can be provided in a safe form. This low-energy or zero-energy sweetener composition is useful as a sweetener composition for people with restricted energy intake, such as obese and diabetic patients, or those who need or desire a diet.

[Food and drink composition]
The Luo Han Guo extract of the present invention can be contained in a food or drink composition.

The Luo Han Guo extract of the present invention is a sweetener that replaces general sucrose, and is used for foods and drinks such as cooked dishes, Western confectionery, Japanese confectionery, beverages (including soft drinks and alcoholic beverages), dairy products, and seasonings. Suitable for use in compositions. Here, the food and drink composition includes not only general foods and drinks but also health foods, special-purpose foods (foods for specified health use) and the like. The Luo Han Guo extract of the present invention is stable, does not cause turbidity, browning, or coloring, and is particularly useful when it is contained in a food or drink composition. These may be supplements such as vitamin C agent, vitamin E agent, vitamin EC agent, vitamin-containing health agent, calcium agent and the like.

The food and drink composition of the present invention may further contain food and drink ingredients commonly used in the art. Examples of food and drink ingredients include lactose, dextrose, sucrose, sorbitol, mannitol, apple fiber, soybean fiber, meat extract, black vinegar extract, gelatin, corn starch, honey, animal and vegetable fats and oils, polysaccharides, grains, vegetables, fruits and vegetables, fruits. , Meat, eggs, dairy products, seaweed, etc. and their processed products. The composition of the present invention may contain one or more of these food ingredients.

The food and drink composition of the present invention may further contain one or more components such as abundant agents, emulsifiers, suspending agents, antioxidants, preservatives, sweeteners and flavoring agents, in addition to the above-mentioned food raw materials. .. In addition, other components including water-soluble vitamins and oil-soluble vitamins may be further contained. Appropriate ingredients that do not interfere with the physical properties and taste of Luo Han Guo extract can be easily selected.

The food and drink composition of the present invention is produced by a method well known in the art. For example, it may be produced by a method including a step of adding a pure Luo Han Guo glycoside having an action, or by a method including a step of adding a composition containing a pure Luo Han Guo glycoside having an action. You may.

In the food and drink composition containing the Rakan fruit extract of the present invention, the dry mass of mogloside V contained in the composition is typically 0.001% by mass to 70% by mass, preferably 0.002% by mass. % To 65% by mass, more preferably 0.003% by mass to 60% by mass, still more preferably 0.004% by mass to 50% by mass, still more preferably 0.005% by mass to 40% by mass. .. Most preferably, it is 0.004% by mass to 30% by mass.

The food and drink composition of the present invention can be produced by a method known to those skilled in the art. Those skilled in the art can select an appropriate production method according to the form and type of the food and drink composition. Here, the Luo Han Guo extract can be incorporated into the food and drink composition by any method.

[Pharmaceutical composition]
The Luo Han Guo extract of the present invention can be contained in a pharmaceutical composition. The Luo Han Guo extract itself can be prepared as the active ingredient of the pharmaceutical composition, and the Luo Han Guo extract can be used as a sweetener together with another active ingredient. The pharmaceutical composition may further comprise pharmaceutically acceptable raw materials. Examples include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose derivatives, tragacanth, gelatin, syrup, methyl hydroxybenzoate, talc. , Magnesium stearate, water, mineral oil and the like. The pharmaceutical composition of the present invention may contain one or more of these raw materials.

The pharmaceutical composition of the present invention contains abundant agents, emulsifiers, suspending agents, antioxidants, preservatives, sweeteners and flavoring agents other than Luo Han Guo extract, and water-soluble vitamins and oil-soluble vitamins. It may contain one or more of the other ingredients.

The form of the pharmaceutical composition of the present invention is not particularly limited. Examples of the forms of the pharmaceutical composition of the present invention include the forms of oral compositions such as tablets, pills, powders, syrups, emulsions, liquids, suspensions, gelatin capsules; and the forms of sprays. Can be mentioned. Pharmaceutical compositions in the form of sprays and the like can also be administered by routes such as intranasally. The oral composition may be a pharmaceutical composition such as a vitamin C agent, a vitamin E agent, a vitamin EC agent, a vitamin-containing health agent, and a calcium agent, as well as an oral composition for treating or preventing various diseases.

In the pharmaceutical composition containing the Rakan fruit extract of the present invention, the dry mass of mogloside V contained in the composition is typically 0.001% by mass to 10% by mass, preferably 0.002% by mass. ~ 5% by mass, more preferably 0.003% by mass to 3% by mass, still more preferably 0.004% by mass to 1% by mass, still more preferably 0.005% by mass to 0.5% by mass. is there. Most preferably, it is 0.006% by mass to 0.3% by mass.

The pharmaceutical composition of the present invention can be produced by a method known to those skilled in the art. One of ordinary skill in the art can select an appropriate production method depending on the form and type of the pharmaceutical composition. Here, the Luo Han Guo extract can be incorporated into the pharmaceutical composition by any method.

[Cosmetic composition]
The Luo Han Guo extract of the present invention can be contained in a cosmetic composition. The Luo Han Guo extract itself can be prepared as an active ingredient in the cosmetic composition, or the Luo Han Guo extract can be used as a sweetener together with another active ingredient. The cosmetic composition can be a medicated cosmetic product. The cosmetic composition may further include other raw materials. Other raw materials include, but are not limited to, surfactants, water, mineral oil and the like. The cosmetic composition of the present invention may contain one or more of these raw materials.

Examples of the cosmetic composition include, but are not limited to, lipstick, toothpaste and the like.

The cosmetic composition of the present invention includes emulsifiers, suspending agents, antioxidants, preservatives, sweeteners and fragrances other than Luo Han Guo extract, and other components including water-soluble vitamins and oil-soluble vitamins. Can include one or more of.

The form of the cosmetic composition of the present invention is not particularly limited. Examples of the form of the cosmetic composition of the present invention include a form such as a spray agent. Examples include, but are not limited to, medicated cosmetics and medicated dentifrices.

In the cosmetic composition containing the Rakan fruit extract of the present invention, the dry mass of mogloside V contained in the composition is typically 0.001% by mass to 10% by mass, preferably 0.002% by mass. % To 5% by mass, more preferably 0.003% by mass to 3% by mass, still more preferably 0.004% by mass to 1% by mass, still more preferably 0.005% by mass to 0.5% by mass. Is. Most preferably, it is 0.006% by mass to 0.3% by mass.

The cosmetic composition of the present invention can be produced by a method known to those skilled in the art. Those skilled in the art can select an appropriate production method depending on the form and type of the cosmetic composition. Here, the Luo Han Guo extract can be incorporated into the cosmetic composition by any method.

[Oral care preparation]
The Luo Han Guo extract of the present invention can be contained in an oral care preparation. The Luo Han Guo extract itself can be prepared as an active ingredient in the cosmetic composition, or the Luo Han Guo extract can be used as a sweetener together with another active ingredient. Oral care formulations may further include other ingredients. Examples include surfactants, water, mineral oil and the like. The oral care preparation of the present invention may contain one or more of these raw materials.

The oral care preparation of the present invention further comprises emulsifiers, suspending agents, antioxidants, preservatives, sweeteners and flavors other than Luo Han Guo extract, and other water-soluble vitamins and oil-soluble vitamins. It may contain one or more of the ingredients.

The form of the oral care preparation of the present invention is not particularly limited. Examples of the form of the oral care preparation of the present invention include a form such as a spray agent; a mouth refreshing agent (for example, a throat refreshing agent, a stomach refreshing agent); and medicated dentifrices. In the present specification, the oral care preparation includes both a composition used as a quasi-drug and a composition used as a newly designated quasi-drug.

In the oral care preparation containing the Rakan fruit extract of the present invention, the dry mass of mogloside V contained in the preparation is typically 0.001% by mass to 10% by mass, preferably 0.002% by mass or more. It is 5% by mass, more preferably 0.003% by mass to 3% by mass, further preferably 0.004% by mass to 1% by mass, and even more preferably 0.005% by mass to 0.5% by mass. .. Most preferably, it is 0.006% by mass to 0.3% by mass.

The oral care preparation of the present invention can be produced by a method known to those skilled in the art. Those skilled in the art can select an appropriate production method according to the form and type of the oral care preparation. Here, the Luo Han Guo extract can be incorporated into the oral care preparation by any method.

Next, the present invention will be specifically described with reference to Examples, but the present invention is not limited to the following Examples.

(Comparative Examples 1 to 8, Reference Examples 1 and 2)
Luo Han Guo fruit (68.5 kg) was added with warm water (60 ° C., 700 L) as a solvent, and the crude product (over 1 mm) was filtered from the Luo Han Guo hot water extract obtained by maintaining heating at 95 ° C. for 60 minutes. The filtrate was spray-dried as it was (Comparative Examples 1 and 2). The temperature of the filtrate at this time was 88 ° C. Further, the filtrate is poured into a column filled with a synthetic adsorption resin, water is passed through the column, and then the concentration of ethyl alcohol is gradient from a low concentration to a high concentration, and mogroside V is present in the column liquid. The fraction was collected, and the concentrated solution in which ethyl alcohol was collected by a concentrator was spray-dried (Comparative Examples 3 to 7). Further, the concentrated solution in which the ethyl alcohol is recovered is poured into a column filled with an ion exchange resin, water is passed through the column, the fraction in which mogroside V is present in the column passing through the column is collected, and water is mixed by a concentrator. The recovered concentrate was spray-dried (Comparative Example 8). In addition, the mixture was poured into a column filled with silica gel surface-modified with octadecylsilyl, water and ethyl alcohol were passed through the column, and then the fraction containing mogroside V in the column was collected and water was used by a concentrator. And the concentrated solution in which the ethyl alcohol was recovered was spray-dried (Reference Example 1).

The spray-dried product of the Luo Han Guo extract obtained above is dissolved in 75v / v% acetonitrile so as to be 10% by mass, and a fraction from which only mogroside V can be obtained is collected using a preparative column (Nucleosil-5C18). Then, a freeze-dried product (Reference Example 2) was obtained.

(Examples 1 to 4)
In order to produce Luo Han Guo extract for the purpose of reducing insoluble matter and increasing clarity, it was placed in a 100 L stainless mesh basket for a general stainless steam jacket extractor (maximum capacity 1,000 L). Luo Han Guo fruit (53.8 kg) and warm water (60 ° C., 550 L) were added, and heating and stirring were started. Extraction was performed for 60 minutes after the liquid temperature reached 95 ° C. or higher. Here, the temperature of the liquid during extraction was monitored, and 95 ± 5 ° C. was maintained for 60 minutes. After extracting with hot water, heating was stopped, and the stainless mesh basket was pulled up to separate Luo Han Guo and the extract (liquid temperature 96 ° C.). When the stirring was stopped and the liquid was sent to the stock tank, it was passed through a Wy-type strainer (opening 1 mm). The liquid temperature at this time was 88 ° C. Then, a crude extract (liquid temperature 39 ° C., sugar content about 3%, liquid turbidity) collected in a stock tank was obtained through a stainless steel liquid feed pipe coated with copper heat radiating fins. The time required to reach the liquid temperature of 39 ° C. through the liquid feeding pipe was within 65 minutes from the time when the extraction was started at 95 ° C. Heat-denatured aggregated proteins, gelled vegetable pectin, dietary fiber, etc. were separated by a continuous centrifuge. The crude extract after centrifugation (liquid temperature 33 ° C., sugar content about 3%, liquid clarification) was concentrated and spray-dried to remove water to obtain 1,573 g of Luo Han Guo extract with improved clarity (liquid clarification). Example 1).

The crude extract after centrifugation obtained by the same method as above is passed through a cylindrical container made of explosion-proof stainless steel filled with synthetic adsorption resin HP20 (Mitsubishi Chemical), and continuously used with ethyl alcohol for food production. A commonly known step of purifying the sweetening component in the Rakan fruit crude extract was added. This solution was concentrated to remove ethyl alcohol, and water was removed by spray-drying to obtain 538 g of Luo Han Guo extract (purified synthetic adsorption resin) having improved clarity (Example 2).
In addition, a generally known step of purifying the crude extract after centrifugation obtained by the same method as above using an anion exchange resin is added to the Luo Han Guo extract purified using a synthetic adsorption resin. did. After concentrating this solution, 264 g of Luo Han Guo extract (purified with anion exchange resin) having improved clarity was obtained by removing water by spray drying (Example 3).

Further, the crude extract after centrifugation obtained by the same method as above is concentrated on the Luo Han Guo extract purified using an anion exchange resin, and then purified again using an anion exchange resin, which is equivalent to the above. The process was repeated. After concentrating the solution obtained in this step, 200 g of Luo Han Guo extract (double-purified anion exchange resin) having improved clarity by removing water by spray drying was obtained (Example 4).

Ten kinds of Luo Han Guo extracts were prepared by the above method, and the degree of purification was "low" (mogroside V content 0.01% by mass or more and less than 20% by mass) depending on the difference in purification degree (mogroside V content). Classification was performed as "medium" (mogroside V content of 20% by mass or more and less than 60% by mass) and degree of purification "high" (mogroside V content of 60% by mass or more and less than 100% by mass) (Comparative Examples 1 to 1). 8. Reference Examples 1 and 2, Examples 1 to 4).

(Test Example 1: Measurement of mogroside V)
Mogroside V was measured by high performance liquid chromatography under the conditions shown below.

(Test Example 2: Detection of insoluble components)
A comparison was made on the insoluble matter contained in the Luo Han Guo extract. Each Luo Han Guo extract was dried at 105 ° C. to a constant weight, the dry weight was determined, and 1 g of the dry weight was weighed (weighed weight value T). This is placed in a 15 mL centrifuge tube (BioLite 15 mL Clinical Centrifuge Tubes; manufactured by Thermo Fisher Scientific) (empty weight value b), and 9 g of distilled water (water temperature 20 to 25 ° C.) is added. In a vortex mixer, suspend the powder until it becomes uniform throughout the aqueous solution, allow it to stand, and then centrifuge at 4,000 rpm for 5 minutes (desktop centrifuge H-36α; manufactured by Kokusan Co., Ltd.) to force insolubilization. Settle things. Then, only the supernatant is removed, and 9 g of distilled water is newly added and suspended by pipetting. Centrifugation was carried out in the same manner, the supernatant was removed, and only the precipitate obtained at this time was frozen at −20 ° C. for 12 to 24 hours (freezer SCR-SK231; manufactured by Sanyo Co., Ltd.). After freezing, it was freeze-dried (freeze-vacuum dryer VD-800R; manufactured by Titec Co., Ltd.) and weighed (weight value c for each centrifuge tube after freeze-drying). For mass measurement in this test, record up to 4 digits after the decimal point using a precision balance (ML104T / 00; manufactured by METTLER TOLEDO). Calculate the proportion of insoluble matter using the calculation formula shown in Equation 1 for each recorded mass value. The same operation was performed three times, and the average value was calculated to obtain the value of the insoluble component (Table 3).
Insoluble matter ratio (%) = (cc) / T × 100 (Equation 1)

(Test Example 3: Evaluation of clarity of aqueous solution)
A comparison was made on the turbidity of the aqueous solution produced by the Luo Han Guo extract. Each Luo Han Guo extract was dried at 105 ° C. to a constant weight, the dry weight was determined, and 1 g of the dry weight was weighed. This was placed in a 15 mL centrifuge tube, and 9 g of distilled water (water temperature 20 to 25 ° C.) was added. The powder was mixed with a vortex mixer until the powder was homogenized throughout the aqueous solution. This mixing operation was completed within 1 minute. After the sample was prepared, the measurement was performed immediately so as not to cause precipitation or precipitation. For the measurement, 3 mL of the liquid was filled in a disposable cell (optical path length 10 mm, optical path width 10 mm), and the absorbance at 660 nm was measured using an ultraviolet-visible spectrophotometer (UV2450; manufactured by Shimadzu Corporation). Distilled water was used as the blank solution. The absorbance was measured by adjusting each Luo Han Guo extract so that it was in the range of 0.100 to 1.000 according to Lambert-Beer's law. The obtained absorbance value was corrected to the absorbance value at 10% by mass of the Luo Han Guo extract in consideration of the amount of the sample weighed using the calculation formula shown in Equation 2. The same operation was performed three times to obtain the average value (Table 3).

Absorbance of sample Ab
Absorbance of blank Bab
Weighing amount (g) M
Absorbance = (Aab-Bab) x (1 / M) (Equation 2)

As a result of examining insoluble substances contained in the Luo Han Guo extracts having different degrees of purification and which are insoluble in water, it was found that they are widely contained in the Luo Han Guo extracts of the comparative examples. In particular, those with a lower degree of purification tended to contain more insoluble matter, and those with a higher degree of purification tended to contain less insoluble matter. A maximum of 12% or more of insoluble matter was contained in the Luo Han Guo extract. As a result of measuring the absorbance at 660 nm, the Luo Han Guo extract of Comparative Example having a large amount of insoluble matter tended to be more turbid, and the Luo Han Guo extract of Example had a low absorbance.

Further, in Examples 1 to 4, as a result of measuring the absorbance of the Luo Han Guo extract obtained by the clarification treatment, the absorbance was lower than that of the Comparative Example containing the same amount of mogroside V, and the turbidity was very small. ..

That is, when the content of mogroside V, which is a measure of the degree of purification of Luo Han Guo extract, is less than 70% by mass and the ratio of insoluble matter to the mass is less than 2% by mass, turbidity (absorbency 660 nm). : Less than 0.3) and it was judged that the clarity was high.

When the aqueous solution of Luo Han Guo extract was visually observed, fine particles could be visually confirmed as suspended matter in the aqueous solution of the comparative example, especially immediately after the adjustment. Further, the aqueous solution of the comparative example could be visually confirmed as deposits and precipitates on the wall surface and bottom of the container with the passage of time. The lower the degree of purification of Luo Han Guo extract, the more of these insoluble substances could be confirmed, and it was not a clear solution.

On the other hand, in the aqueous solution of Example, no suspended matter, deposits or precipitates were visually observed in the aqueous solution after the adjustment, and the solution was a clear solution.

As described above, although there is a difference between the compositions of Examples and Comparative Examples in the absorbance measurement, a large difference was visually observed between Comparative Examples and Examples.

(Test Example 4: Visual evaluation)
A detailed evaluation of the visual impression of the Luo Han Guo extract was performed. For 11 subjects, visual sensory evaluation was performed by absolute evaluation on the aqueous solution of Comparative Example 1 and the 10% by mass aqueous solution of Example 1. As for the evaluation contents, we asked questions that can be answered with "YES" and "NO" about 4 items related to suspended matter, microorganisms, motivation, and clarification. In the questionnaire results, the number of people who answered "YES" was counted (Table 4).

As a result, in Comparative Example 1, all the subjects answered that there was a floating substance. Despite the fact that the general viable cell count was below the standard at which growth was not observed, the majority of the subjects inferred the growth of microorganisms only by the appearance of the aqueous solution. In addition, the majority of the subjects answered that they were resistant to eating. On the other hand, in Example 1, it was judged that there was no resistance to feeding because there was no floating matter when used as a sweetener and an impression of clearness was obtained.

(Test Example 5: Sensory evaluation of Luo Han Guo extract containing no water-insoluble component)
In order to examine the influence of the water-insoluble component on the sensory evaluation items such as the physical properties and taste of the Luo Han Guo extract, only the insoluble matter in the Luo Han Guo extract was prepared by the above-mentioned method, and the Luo Han Guo extract of Example 2 was used. On the other hand, an insoluble component was added to obtain an adjusted amount of Luo Han Guo extract (Examples 5 to 7).

In order to verify the effect, 13 subjects (AM) who did not know the background about the fact that the Luo Han Guo extract contained a water-insoluble component were asked to use the screw tube No. A pair of 10 mass% aqueous solutions of two types (reference product: Comparative Example 1, target products: Comparative Examples 1 to 3 and Examples 2, 5 to 7) adjusted in 5 (manufactured by Maruem) and a score table are presented. He asked, "Is there any difference between the floating substances in these two Luo Han Guo extracts?" And conducted a visual evaluation. The answers to the questions were evaluated using a three-level score table (“0: equivalent”, “+1: better”, and “-1: worse”) (Table 5). At the time of totaling this visual evaluation, the numerical value obtained by calculating the evaluation of the target product with respect to the standard product and summing the numerical values of 13 persons was used as the discrimination score. For each subject, an interval of 3 hours or more was provided.

Regarding the insoluble matter in the Luo Han Guo extract, the majority of the subjects evaluated that there was a clear improvement by visual evaluation "when the water-insoluble component in the Luo Han Guo extract was less than 5% by mass".

(Test Example 6: Luo Han Guo extract protein, polyphenol quantification)
Protein quantification was performed to verify the water-insoluble components of Luo Han Guo extract. A comparison was made regarding the protein content. 0.1 g of each Rakan fruit extract was weighed, placed in a 15 mL centrifuge tube, 0.9 g of 0.9% sodium chloride aqueous solution was added, and suspended in a vortex mixer until the powder was homogenized throughout the aqueous solution. .. 0.1 g of this solution was weighed, and 9.9 g of a 0.9% aqueous sodium chloride solution was added (total protein content).

In addition, weigh 0.1 g of each Rakan fruit extract, put it in a 15 mL centrifuge tube, add 0.9 g of 0.9% sodium chloride aqueous solution, and hang with a vortex mixer until the powder is homogenized throughout the aqueous solution. It became cloudy. After standing, centrifugation was performed at 4,000 rpm for 5 minutes to forcibly settle the insoluble matter. Then only the supernatant was carefully removed. The precipitate was suspended and dissolved in 1.0 g of a 0.9% aqueous sodium chloride solution (insoluble protein content).

These were performed using a BCA protein assay (Thermo Fisher Science). The Working requirement was adjusted by mixing Solution A and Solution B at a ratio of 50: 1. A 50 microliter sample was mixed with 1 milliliter of Working reagent. It was warmed at 37 ° C. for 30 minutes and then allowed to stand until it returned to room temperature. The absorbance at a wavelength of 562 nm was measured. At the same time, a calibration curve (R ² ≧ 0.999) was prepared using bovine serum albumin as a standard substance. The concentration of the sample was appropriately adjusted so as to be within the range of the calibration curve. The mass in the insoluble matter was calculated using the protein concentration in the sample obtained from the calibration curve, and the content in the Luo Han Guo extract was calculated. The same operation was performed three times to obtain the average value (Table 6).

Next, the amount of polyphenol was quantified.
FOLIN-DENIS reagents were prepared. To 188.0 mL of distilled water, 27.73 g of sodium tungstate dihydrate, 5 g of phosphomolybdic acid and 12.5 mL of 85% phosphoric acid were added. The mixed solution was refluxed in a fume hood using a Liebig condenser for 2 hours. After the reflux operation, the mixture was allowed to cool, and after the liquid temperature reached room temperature, the flask was transferred to a 1000 mL volumetric flask and filled up with distilled water up to the marked line. This solution (FOLIN-DENIS REAGENT) was subdivided into a light-shielding container and stored in a cool and dark place until use.

In addition, 0.1 g of each Luo Han Guo extract (Total) and the insoluble component (Ppt) of each Luo Han Guo extract obtained by the method of Test Example 2 were weighed and dissolved or suspended in 1 mL of water. 0.5 mL of FOLIM-DENIS REAGENT and 0.5 mL of the dissolved or suspended sample were added to a 2 mL microtube, and after 3 minutes, a 10 w / v% aqueous sodium carbonate solution was added. Centrifugation was performed 60 minutes later (14,000 G, 5 minutes, room temperature). The absorbance at 760 nm was measured with respect to the supernatant of the sample after centrifugation. Distilled water was used as the blank solution. The absorbance was measured by adjusting the solution so that it was in the range of 0.100 to 1.000 according to Lambert-Beer's law. Similarly, a calibration curve (R ² ≧ 0.999) was prepared using tannic acid as a standard substance. A 0.1 mg / mL tannic acid aqueous solution was prepared and diluted appropriately. The amount of polyphenol in Total and Ppt was determined using the polyphenol concentration in the sample obtained from the calibration curve, and the content in Luo Han Guo extract was calculated. The same operation was performed three times to obtain the average value (Table 6).

Using the values of the total polyphenol (Total) content in the obtained Luo Han Guo extract and the insoluble polyphenol (Ppt) content in the insoluble component, the polyphenol abundance ratio in the sample was calculated using the formula shown in Equation 3. I asked.
Polyphenol abundance = (Ppt / Total) ... (Equation 3)

As a result of quantifying the protein and polyphenol in the Luo Han Guo extract, it was clarified from Comparative Examples 1, 3, 8 and Reference Example 1 that the main component forming the insoluble matter was polyphenol. Many proteins were present in the Luo Han Guo extract, but most of them were not present in the insoluble matter, but were present in the aqueous solution. From Examples 1 and 2, it was found that polyphenols were present in the Luo Han Guo extract but were present in the aqueous solution.

From the results in Table 7, the Luo Han Guo extract composition of the present invention prepared in Examples 1 to 3 has a mogroside content of insoluble components as compared with the Luo Han Guo extract composition prepared by the prior art of Comparative Examples 1 to 3. It was found to be significantly reduced compared to the V content.

(Test Example 7: Sensory evaluation)
The Luo Han Guo extracts prepared in Examples 1 to 3 and Comparative Examples 1 to 3 were subjected to sensory evaluation by 10 skilled subjects (A to J). The sweetness intensity of the aqueous solution of each Luo Han Guo extract was adjusted to be equivalent to that of the sucrose (10 w / v%: control solution) aqueous solution. In the sensory evaluation, five items (bitterness, astringency, habit, backlash, and texture) representing sweetness were evaluated, and the scores were given as follows according to the sensory evaluation criteria.
Compared to sucrose solution
0: Very good 1: Excellent 2: Somewhat good 3: Similar 4: Somewhat inferior 5: Inferior 6: Very inferior

Regarding the definition of taste, Japanese Industrial Standards: JIS Z 8144: 2004 Sensory Evaluation Analysis-Skilled subjects are those who can distinguish and evaluate these according to the terms, and more specifically, bitterness is "when eating coffee or fish guts". "Taste that you can feel with your tongue", astringency is "taste caused by contraction of the tongue mucosa", and habit is "taste that makes you feel unpleasant other than bitterness and astringency". .. Furthermore, the backtracking is "the length of time that the tongue feels when eating (sweetened) sweets and fruits", and the texture is "the quality and quantity of the substance that remains on the tongue." That is. " These items are so good that they are weak or hard to feel. The evaluation point of the sucrose aqueous solution as a control solution was 3 points, indicating that the sweetness was of good quality (Table 8).

Here, in Examples 1, 2 and 3, all the items are close to 2 to 3 points, and the taste is close to that of sucrose, and while feeling sweet, there is no backing, the texture is smooth, and the texture is smooth. Bitterness and astringency were suppressed. Comparative Examples 1, 2 and 3 had a taste inferior to that of sucrose, which was close to 4 to 5 points.

As a result, according to Examples 1 to 3, "when the water-insoluble component in the Luo Han Guo extract is less than 5% by mass", the bitterness, astringency, habit and backing of the Luo Han Guo extract are reduced, and the sweetness is close to that of sucrose. showed that. Furthermore, the discomfort attached to the tongue when it was put into the mouth was reduced, and the texture of the tongue was cleaner than that of sucrose. In each case, it was evaluated that there was a clear improvement and was preferred. It was considered that water-insoluble components other than the sweet component contained in the Luo Han Guo extract deteriorate the taste quality.

(Test Example 8: Consolidation test)
Luo Han Guo of Examples 1 to 3 prepared above in order to investigate the influence of the content of mogroside V and the content of water-insoluble components in the Luo Han Guo extract composition on the time-dependent (moisture absorption) consolidation of the powder. The extract and the Luo Han Guo extract of Comparative Examples 1 to 3 were examined by a consolidation test.

In the solidification test, the solidification characteristics of the powder can be evaluated by storing the powder filled in each packaging material under each condition for a certain period of time. For the packaging material of each Luo Han Guo extract, Unipack (registered trademark) (material: polyethylene, thickness 0.04 mm, manufactured by Japan Co., Ltd.) made of a material that can be used for food was used.

Regarding the evaluation, the degree of consolidation is scored according to the stage (number of consolidation points 0: no consolidation, number of consolidation points 1: extremely soft consolidation occurs / solidification, but cannot be pinched with fingers Number of consolidation points 2: Occurrence of soft consolidation ・ Can be easily pinched with a finger and easily collapses when pressed, Number of consolidation points 3: Occurrence of solid consolidation ・ Can be easily pinched with a finger and pressed Force is required to break, number of consolidation points 4: generation of extremely hard consolidation ・ It is necessary to use both hands to break the consolidation, number of consolidation points 5: occurrence of consolidation that cannot be broken by hand) (Table 9). The same operation was performed three times, and the average value was calculated. I went for up to 35 days. The packaging material used for the evaluation once was discarded because the solidification characteristics changed.

As a result, in Examples 1 to 3, the occurrence of caking was suppressed and the storage stability was improved. It was considered that the water-insoluble component contained in the Luo Han Guo extract reduces the storage stability.

(Test Example 9: Permeability of Luo Han Guo extract)
Screw tube No. An aqueous solution was prepared in 3 (manufactured by Maruem Co., Ltd.) so that the Luo Han Guo extract was 1% by mass, and the permeability was confirmed (Fig. 1: water from left, Comparative Example 3, Example 2, water).

According to Examples 1 to 3, the generation of turbidity in the aqueous solution of Luo Han Guo extract was suppressed, and the clarity was improved. It was considered that water-insoluble components other than the sweet component contained in the Luo Han Guo extract reduce the clarity.

(Composition)
Based on the blending amounts in Table 10, fructose-glucose liquid sugar, Luo Han Guo extract, and drinking water were mixed and filled in a container as -100 parts by mass to prepare a high-concentration solution. For comparison, a high-concentration solution containing a conventional Luo Han Guo extract instead of the product of the present invention was prepared. Hereinafter, the numerical values in the table indicate mass%.

Ten subjects evaluated the prototype high-concentration solution for the degree of turbidity of the solution by visual evaluation and the taste quality by tasting.
As a result of the sensory evaluation, insoluble matter was confirmed in the solution of the comparative food and drink composition 1, and it was judged that the majority of the subjects were turbid, and the sweetness was weak and the subject was astringent. On the other hand, it was found that the food and drink composition 1 using the Luo Han Guo extract of the example had a sweet taste and a weak astringency.

Based on the blending amounts in Table 11, 95 v / v% alcohol, high fructose corn syrup, erythritol, Luo Han Guo extract, and drinking water were mixed and filled in a container as 100 parts by mass to prepare an alcohol solution. For comparison, an alcohol solution containing a conventional Luo Han Guo extract instead of the product of the present invention was prepared.

We asked 10 subjects to evaluate the turbidity of the prototype alcohol solution by visual evaluation and the taste quality by tasting.

As a result of the sensory evaluation, insoluble matter was confirmed in the solution of the comparative food and drink composition 2, and it was judged that the majority of the subjects were turbid, and the sweetness was weak and the astringency was astringent. On the other hand, it was found that the food and drink composition 2 using the Luo Han Guo extract of the example had a sweet taste and a weak astringency.

Based on the blending amount in Table 12, fructose corn syrup, erythritol, lemon juice, Luo Han Guo extract, citric acid, trisodium citrate, flavor, sodium benzoate, and drinking water are mixed and filled in a container as 100 parts by mass. To prepare an acidic solution. In addition, for comparison, an acidic solution containing the Luo Han Guo extract of Comparative Example was prepared instead of Example.

Ten subjects evaluated the sampled acidic solution for the degree of turbidity of the solution by visual evaluation and the taste quality by tasting.
As a result of the sensory evaluation, insoluble matter was confirmed in the solution of the comparative food and drink composition 3, and it was judged that the majority of the subjects were turbid, and the sweetness was weak and the astringency was astringent. On the other hand, it was found that the food and drink composition 3 using the Luo Han Guo extract of the example had a sweet taste and a weak astringency.

In addition, in order to obtain a clear, high-quality, highly clear Luo Han Guo extract, the production time was verified, and the time required for each step in the above-mentioned production method was measured. Regarding the manufacturing time, we used data that standardized the measurement record for one year in order to suppress fluctuations with time. (Table 13, Fig. 2)

Assuming that the production time in the comparative example was 100, the production time was 74 in the example. In the comparative example (conventional technique), since the column purification step required for clarification is not required in the present invention, the production time alone can be reduced by about 26%. Further, in column purification, cleaning / regeneration treatment is required after use, but since it is not included in the production time in Table 13, the man-hours can be further shortened in the examples.

(Test Example 10: Parameters by extraction time)
When obtaining a highly clear Luo Han Guo extract with reduced insoluble matter, we confirmed the difference in physical properties due to the time effect of the temperature load during heating and cooling. Add Luo Han Guo fruit (53.8 kg) and warm water (60 ° C, 550 L) in a 100 L stainless mesh basket to a general stainless steam jacket extractor (maximum capacity 1,000 L), and start heating and stirring. did. Extraction was performed for 60 minutes after the liquid temperature reached 95 ° C. or higher. Here, the temperature of the liquid during extraction was monitored, and 95 ± 5 ° C. was maintained for 60 minutes. After extracting with hot water, heating was stopped, and the stainless mesh basket was pulled up to separate Luo Han Guo and the extract (liquid temperature 96 ° C.). When the stirring was stopped and the liquid was sent to the stock tank, it was passed through a Wy-type strainer (opening 1 mm). (The liquid temperature at this time was 88 ° C.). Then, a crude extract (liquid temperature 39 ° C., sugar content about 3%, liquid turbidity) collected in a stock tank was obtained through a stainless steel liquid feed pipe coated with copper heat radiating fins. The time when the liquid temperature reached 39 ° C. through the liquid feeding pipe was analyzed by changing the time from the time when the extraction was started at 95 ° C. By passing the extract from Luo Han Guo through a Wy-type strainer and adjusting the feed rate and the number of heat-dissipating fins before sending it to the stock tank, the crude extract collected in the stock tank reaches 40 degrees or less. The amount of heat load was changed. The heat load was adjusted under each condition, and then the heat-denatured aggregated protein, gelled vegetable pectin, dietary fiber, etc. were separated by a continuous centrifuge. The crude extract after centrifugation (liquid temperature 33 ° C., sugar content about 3%, liquid clarification) was concentrated and spray-dried to remove water to obtain a Luo Han Guo extract with improved clarification. The results are shown in Table 14. From Example 17 onward, the crude extract was purified using a synthetic adsorbent and an anion exchange resin.

Here, even in the compositions of Comparative Examples 11 and 12 having a relatively low absorbance (660 nm) value, turbidity was clearly generated visually, and the composition was not in a clear state as in the composition of the example. It was.

[Example of composition]
A composition containing the Luo Han Guo extract of the present invention was prepared based on the formulations shown in Tables 15 to 20 below. The numerical value in the table means mass%.

Claims (9)

Luo Han Guo (Siraita grosvenorii) extract,
(1) Mogroside V is contained in an amount of 0.01% by mass to 70% by mass; and (2) a water-insoluble component is contained in an amount of 5% by mass or less.
Furthermore, the insoluble polyphenol content / total polyphenol content is 1/2 or less.
Luo Han Guo extract. The Luo Han Guo extract according to claim 1, wherein the absorbance at a wavelength of 660 nm in a 10 mass% aqueous solution is 0.3 or less. The Luo Han Guo extract according to any one of claims 1 or 2, wherein the total polyphenol content is 0.1% by mass to 20% by mass. The Luo Han Guo extract according to any one of claims 1 to 3, wherein the insoluble polyphenol content is 5% by mass or less. The Luo Han Guo extract according to any one of claims 1 to 4, wherein the total protein content is 5% by mass to 90% by mass. The Luo Han Guo extract according to any one of claims 1 to 5, wherein the insoluble protein content is 3% by mass or less. A food or drink composition containing the Luo Han Guo extract according to any one of claims 1 to 6. A pharmaceutical composition containing the Luo Han Guo extract according to any one of claims 1 to 6. A cosmetic composition or an oral care preparation containing the Luo Han Guo extract according to any one of claims 1 to 6.