Method for diluting and oil-solubilizing ethanol extract of bitter melon leaves

Abstract

To provide a method for diluting, with water, an ethanolic extract of bitter melon (Momordica charantia) leaves, capable of suppressing formation of deposition as much as possible; and to provide a method for making the ethanolic extract of bitter melon leaves oil-soluble.SOLUTION: There is provided a method for diluting an ethanolic extract of bitter melon (Momordica charantia) leaves by adding water to the ethanolic extract of the bitter melon leaves, with a water extract of Nemacystus decipiens added thereto. There is provided also a method for adding an emulsifier and lecithin to the ethanolic extract of bitter melon leaves, with the water extract of Nemacystus decipiens added thereto, and further preparing an emulsion with oil added thereto to make the ethanolic extract of bitter melon leaves oil soluble with the emulsion added in the oil.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a method for diluting and a method for oil solubilizing an ethanol extract of Turraea leaves.

Background Art

[0002] In Japan, the number of patients with allergic diseases has increased dramatically since the period of high economic growth due to factors such as the Westernization of the diet centered on bread and house dust.

[0003] Particularly, gluten allergy has problems such as the antigen being derived from staple foods such as bread-based diets and the antigenicity of gluten as an antigen not being suppressed even by heat treatment. As a result, patients are forced to change the raw materials of staple foods themselves and have an inconvenient life.

[0004] [[ID=2)) If the onset of allergies can be suppressed through daily diet, the burden can be reduced.

[0005] The present inventors have previously proposed a gluten allergy inhibitor in the form of a tablet containing a dried product of an extraction stock solution from Turraea leaves (see Patent Document 1).

[0006] According to such a gluten allergy inhibitor, it is possible to provide a gluten allergy inhibitor using natural products that can suppress the onset of gluten allergy through daily diet.

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0008] By the way, the gluten allergy suppressant mentioned above is made into a solid (tablet-like) form using the dried extract of bitter melon leaves. However, if we want to increase the variety of dosage forms, such as liquid or jelly, it would be simpler to use the liquid extract as is.

[0009] However, when the raw extract of bitter melon leaves is an ethanol extract, there is a problem in that insoluble components precipitate when, for example, diluting it with water to prepare an aqueous gluten allergy inhibitor or attempting to solubilize it with oil to prepare an oil-based gluten allergy inhibitor.

[0010] Furthermore, these problems are not limited to the use of bitter melon leaf ethanol extract as a gluten allergy suppressant; rather, they severely restrict the range of applications of bitter melon leaf ethanol extract, given the various effects of bitter melon leaves that are expected, such as improving dementia.

[0011] The present invention has been made in view of these circumstances, and provides a method for diluting an ethanol extract of bitter melon leaves with water that can suppress the formation of precipitate as much as possible, and a method for oil-solubilizing an ethanol extract of bitter melon leaves. [Means for solving the problem]

[0012] To solve the above conventional problems, the method for diluting bitter melon leaf ethanol extract according to the present invention includes (1) Contains 0.0167 to 16.67 parts by weight of fucose. Added water extract from mozuku seaweed Contains 1 part by weight of momordicin We decided to dilute the ethanol extract of bitter melon leaves with water. Furthermore, in the method for producing a diluted solution of the bitter melon leaf ethanol extract according to the present invention, (2) water was added to the ethanol extract of bitter melon leaves containing 1 part by weight of momordicin, to which 0.0167 to 16.67 parts by weight of water extract of mozuku containing fucose was added, and then water was added to dilute it.

[0013] Furthermore, in the method for solubilizing the ethanol extract of bitter melon leaves according to the present invention, 3 ) Contains 0.0167 to 16.67 parts by weight of fucose. Added water extract from mozuku seaweed Contains 1 part by weight of momordicin Emulsifier added to ethanol extract of bitter melon leaves (However, the final concentration after solubilization in oil corresponds to an amount of 0.1 to 50 mg / ml) lecithin (Includes.)It was added, and further oil was added to prepare an emulsion, and the emulsion was added to the oil. Furthermore, in the method for producing an oil-soluble solution of bitter melon leaf ethanol extract according to the present invention, (4) an emulsifier (however, containing an amount of lecithin equivalent to 0.1 to 50 mg / ml at the final concentration after oil solubilization) is added to an ethanol extract of bitter melon leaf containing 1 part by weight of momordicin, to which 0.0167 to 16.67 parts by weight of water extract of mozuku containing fucose is added, and then oil is added to prepare an emulsion, which is then added to the oil.

[0014] Also, in the present invention, ( 5 ) A water extract of mozuku seaweed containing 0.0167 to 16.67 parts by weight of fucose is added to an ethanol extract of bitter melon leaf containing 1 part by weight of momordicin. As a precipitation inhibitor for the ethanol extract of Turraea leaves use It was used.

Effects of the Invention

[0015] According to the method for diluting the ethanol extract of Turraea leaves according to the present invention or method of producing a diluted solution According to which, Contains 0.0167 to 16.67 parts by weight of fucose. Since water is added to the ethanol extract of Turraea leaves to which the water extract of mozuku is added and diluted, a method for diluting the ethanol extract of Turraea leaves with water that can suppress precipitation generation as much as possible Contains 1 part by weight of momordicin Can be provided. or method of producing a diluted solution

[0016] Method for producing oil-soluble liquids Also, according to the method for solubilizing the ethanol extract of Turraea leaves according to the present invention Contains 0.0167 to 16.67 parts by weight of fucose. According to which, Contains 1 part by weight of momordicin Since the emulsifier (However, the final concentration after solubilization in oil corresponds to an amount of 0.1 to 50 mg / ml) Lecithin (Includes.) Is added to the ethanol extract of Turraea leaves to which the water extract of mozuku is added, and further oil is added to prepare an emulsion, and the emulsion is added to the oil, a method for solubilizing the ethanol extract of Turraea leaves that can suppress precipitation generation as much as possible or method for producing oil-soluble liquids Can be provided.

[0017] A water extract of mozuku seaweed containing 0.0167 to 16.67 parts by weight of fucose is added to an ethanol extract of bitter melon leaf containing 1 part by weight of momordicin. use Also, Since it is used as a precipitation inhibitor for the ethanol extract of Turraea leaves When diluting the ethanol extract of Turraea leaves with water or solubilizing the ethanol extract of Turraea leaves, precipitation generation can be suppressed as much as possible.

Modes for Carrying Out the Invention

[0018] ​​This invention provides a method for diluting a bitter melon leaf ethanol extract. Here, the bitter melon leaf ethanol extract is an ethanol extract from bitter melon (momordica charantia), and contains extracts from the leaves and stems of bitter melon as active ingredients. The main components of this extract are momordicin, charantin, and corosolic acid, but other components may also be present.

[0019] Furthermore, the leaves of other plants in the genus Momordica (i.e., the genus Momordica), such as Momordica gracilis, Trichosanthes cucumeroides, and Momordica kacrole, may also be used as raw materials for the Momordica leaf ethanol extract.

[0020] Furthermore, the leaves and stems of the bitter melon used as raw materials may be harvested from bitter melon plants that have been cultivated with bud formation, or from bitter melon plants that have been cultivated without bud formation. However, in bitter melon plants that have been cultivated without bud formation, the active ingredients can be concentrated in the leaves and stems, so leaves and stems harvested from bitter melon plants that have been cultivated without bud formation are more preferable.

[0021] Extraction of bitter melon leaves with ethanol can be done at room temperature or with hot ethanol. For hot ethanol extraction, the temperature can be, for example, 30-95°C.

[0022] The ethanol used for extraction may contain water, for example, if it is diluted with water. The acceptable amount of water is greater than 0 w / w% but generally 50 w / w% or less.

[0023] Furthermore, a characteristic of the method for diluting the bitter melon leaf ethanol extract according to the present invention is that, prior to dilution with water, a water extract of mozuku seaweed is added to the bitter melon leaf ethanol extract.

[0024] The mozuku seaweed used as the raw material for the water extract of mozuku is preferably Okinawa mozuku (Cladosiphon okamuranus), a species of brown alga belonging to the family Chondrophyllaceae in the order Cyanobacteriales.

[0025] Mozuku water extract is an extract obtained by washing mozuku and extracting it with water or hot water, either untreated or after a predetermined treatment, and is rich in fucoidan and fucose. Examples of predetermined treatments include acid treatment and base treatment (alkali treatment).

[0026] The amount of mozuku water extract added to the bitter melon leaf ethanol extract should be an amount that achieves the objective, that is, an amount that suppresses the precipitation of insoluble components, and is not particularly limited. It can be adjusted as appropriate during use. However, since this application makes a new proposal in this regard, the amount of mozuku water extract added to the bitter melon leaf ethanol extract will be discussed later.

[0027] Thus, when a water extract of mozuku seaweed is added to a bitter melon leaf ethanol extract, the precipitation of insoluble components can be suppressed even when diluted with water, and a stable aqueous formulation with a low amount of precipitate can be prepared.

[0028] The present invention also provides a method for oil-solubilizing a bitter melon leaf ethanol extract. When oil is added to a bitter melon leaf ethanol extract, precipitation of insoluble components occurs.

[0029] Therefore, in the method for oil-solubilizing the ethanol extract of bitter melon leaves according to the present invention, an emulsifier and lecithin are added to the ethanol extract of bitter melon leaves to which a water extract of mozuku seaweed has been added, and then oil is added to prepare an emulsion, which is then added to the oil.

[0030] Here, the amount of mozuku water extract added to the bitter melon leaf ethanol extract required for oil solubility should be an amount that achieves the objective, that is, an amount that suppresses the precipitation of insoluble components, and is not particularly limited. It can be adjusted as appropriate during use. However, this point also represents a new proposal in this application, which will be discussed later.

[0031] The bitter melon leaf ethanol extract, to which a water extract of mozuku seaweed has been added, is then subjected to the addition of an emulsifier and lecithin.

[0032] The emulsifier can be any commercially available emulsifier, such as lecithin-based or fatty acid glyceryl-based emulsifiers, as long as its HLB value is around 1 to 16.

[0033] The amount of emulsifier added should be sufficient to emulsify or solubilize the insoluble components, and will vary depending on the type of emulsifier used. For example, compared to a bitter melon leaf ethanol extract to which a water extract of mozuku seaweed has been added... The final concentration after oil solubilization is Generally, concentrations range from 0.1 to 50 mg / ml. The amount It is common practice to add it this way.

[0034] For lecithin, for example, lecithin with a high phosphatidylcholine content or hydrogenated lecithin can be used.

[0035] The amount of lecithin added should be sufficient to emulsify or solubilize the insoluble components, and will vary depending on the type of lecithin used. For example, in comparison to a bitter melon leaf ethanol extract to which a water extract of mozuku seaweed has been added... The final concentration after oil solubilization is Generally, concentrations range from 0.1 to 50 mg / ml. The amount It is common practice to add it this way.

[0036] Next, oil is added to the mozuku water extract, emulsifier, and lecithin-containing bitter melon leaf ethanol extract to emulsify it.

[0037] The oil used for emulsification can be appropriately selected depending on the product utilizing the bitter melon leaf ethanol extract. For example, if the product is a food product, it is desirable to select an oil that is suitable for consumption, and if it is a pharmaceutical product, it is desirable to select an oil that is suitable for use at a pharmaceutical level. To give a specific example, for food products, olive oil or MCT (medium-chain triglyceride) oil can be suitably used.

[0038] The amount of oil used for emulsification is not particularly limited, but the total amount of mozuku water extract, emulsifier, and lecithin added to the bitter melon leaf ethanol extract can be greater than or equal to the amount that can be emulsified.

[0039] Then, by adding the emulsion prepared in this way to the dissolving oil, the oil-soluble form of the bitter melon leaf ethanol extract is achieved. The emulsifying oil and the dissolving oil may be the same oil, or they may be different oils. Furthermore, the emulsifying oil and the dissolving oil can be added in separate steps, but they can also be added in a single step that serves both emulsification and dissolution.

[0040] Furthermore, considering the dilution method and oil-solubilization method of the bitter melon leaf ethanol extract according to this embodiment from another perspective, it can be said that this application also provides a method for using a water extract of mozuku seaweed as a precipitation inhibitor for the bitter melon leaf ethanol extract.

[0041] By the way, as mentioned earlier, the amount of mozuku water extract added to the bitter melon leaf ethanol extract, which is necessary for dilution with water or oil solubility, is not particularly limited as long as it is an amount that can suppress the precipitation of insoluble components.

[0042] However, determining the amount of mozuku water extract needed to suppress precipitate formation may require adjustment depending on the concentration of the bitter melon leaf ethanol extract and the concentration of the mozuku water extract, and this determination can be somewhat complicated.

[0043] Furthermore, the specific mechanism by which mozuku water extract inhibits precipitation requires further research, and since the precipitation-inhibiting component in the mozuku water extract is currently unknown, its content cannot be used as an indicator. Similarly, the mechanism by which precipitation occurs when bitter melon leaf ethanol extract is diluted or dissolved in oil is currently unknown, making it difficult to determine what criteria to use when deciding on the amount of mozuku water extract to add.

[0044] In this regard, the present invention determines the amount of mozuku water extract added to the bitter melon leaf ethanol extract by mixing the bitter melon leaf ethanol extract and the mozuku water extract, using momordicin derived from the bitter melon leaf ethanol extract and fucose derived from the mozuku water extract as reference components, so that momordicin and fucose are in a predetermined ratio. It is also possible to use total momordicin as a reference component or individual momordicins as a reference component. For example, in this embodiment, the amount of mozuku water extract added to the bitter melon leaf ethanol extract is characterized in that the amount of fucose contained in the mozuku water extract is 0.01 to 1 mg for every 1 mg of momordicin I contained in the bitter melon leaf ethanol extract.

[0045] While this is merely speculation at this point, and for the sake of understanding the present invention, the inventors believe that momordicin is not directly involved in precipitation formation during dilution or oil solubility, nor is fucose directly involved in suppressing precipitation formation.

[0046] However, based on the experience gained through our diligent research, we have found that by using the amounts of momordicin and fucose as indicators and adjusting the amounts of both within the above range, precipitation during dilution and oil solubilization can be effectively suppressed, which is extremely useful in eliminating the complexity associated with adjusting the above-mentioned amounts.

[0047] As an example, the amount of mozuku water extract added to the above bitter melon leaf ethanol extract results in a momordicin I concentration of 0.0001-0.005 w / w% in the bitter melon leaf ethanol extract and a fucose concentration of 0.01-1 w / w% in the mozuku water extract. This combination is expected to suppress precipitation during dilution or oil solubilization, where the final concentration of the bitter melon leaf ethanol extract is less than 100 w / w% and 0.1 w / w% or higher.

[0048] The following describes in more detail the dilution method and oil-solubilization method of the bitter melon leaf ethanol extract according to this embodiment, referring to experimental examples that take into account the preparation process of bitter melon leaf extract, the preparation process of mozuku water extract (hereinafter also referred to as mozuku water extract), and the use of mozuku water extract as a precipitation inhibitor for the bitter melon leaf ethanol extract.

[0049] [1. Preparation of ethanol extract of bitter melon leaves] 100g of Momordica charantia leaves were placed in a 3L beaker, and 2kg of 95 vol% ethanol at approximately 20°C was added. The extraction was carried out by stirring for 1 to 4 hours. The Momordica charantia leaves were then removed by filtration, and the ethanol was removed to obtain an ethanol extract of Momordica charantia leaves. The concentration of momordicin I in the ethanol extract of Momordica charantia leaves prepared in this way was 0.03 mg / g.

[0050] [2. Preparation of Mozuku Water Extract] 100g of mozuku seaweed (Cladosiphon okamuranus) was placed in a 1L beaker, 500g of water was added, and the mixture was heated at 90°C for 3 hours while stirring well. After that, the mozuku was removed, and the resulting solution was used as the mozuku water extract. The fucose concentration of the mozuku water extract obtained in this way was 5mg / g.

[0051] [3. Water solubility test] Next, a water solubility test was conducted using the ethanol extract of bitter melon leaves described above. Specifically, the test examined how the precipitation of insoluble components changed when water was added to the ethanol extract of bitter melon leaves, depending on whether or not mozuku seaweed extract was added.

[0052] The test involved visually comparing two samples: Comparative Sample 1, which was water-soluble without the addition of mozuku water extract, and Confirmation Sample 2, which was water-soluble with the addition of mozuku water extract, to examine the presence or absence of insoluble components.

[0053] Comparative sample 1 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker and adding 95 g of water to make a 5 w / w% aqueous solution.

[0054] For confirmation sample 2, 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf was dispensed into a 200 ml beaker, 0.015 g (0.075 mg as fucose) of mozuku seaweed extract was added, and then 94.985 g of water was added to make a 5% aqueous solution. Table 1 shows the results of the water solubility test. [Table 1]

[0055] As can be seen from the results of comparative sample 1 shown in sample No. 1 in Table 1, when the ethanol extract of bitter melon leaves is added to an aqueous solution, insoluble components precipitate. However, as can be seen from the results of confirmation sample 2 shown in sample No. 2, when a water-extracted mozuku seaweed extract was added to the ethanol extract of bitter melon leaves before adding water, no precipitation of insoluble components was observed.

[0056] It is thought that the mozuku water extract contributed to the solubilization of insoluble components, but it has never been previously considered that mozuku water extract would have solubilizing properties.

[0057] Further experiments revealed that the optimal mixing ratio of mozuku seaweed extract to bitter melon leaf ethanol extract is between 0.01% and 10%. Below this range, there is no effect, and above this range, insoluble components precipitate, which was discovered through additional experiments.

[0058] These findings demonstrate that the method for diluting the bitter melon leaf ethanol extract according to this embodiment is a method of diluting the bitter melon leaf ethanol extract with water that can suppress the precipitation of insoluble components as much as possible.

[0059] [4. Oil-Solubilization Test] Next, an oil-solubility test was conducted using the ethanol extract of bitter melon leaves described above. Specifically, the precipitation of insoluble components when oil was added to the ethanol extract of bitter melon leaves was examined to see how it changed depending on whether or not emulsifiers, lecithin, and mozuku seaweed extract were added.

[0060] The test involved visually comparing three comparative samples: Comparative Sample 3, which attempted oil solubility without adding any emulsifier, lecithin, or mozuku water extract; Comparative Sample 4, which attempted oil solubility with the addition of an emulsifier but without lecithin or mozuku water extract; and Comparative Sample 5, which attempted oil solubility with the addition of an emulsifier and lecithin but without mozuku water extract. These were then compared with a confirmation sample 6, which contained all three ingredients: emulsifier, lecithin, and mozuku water extract. The presence or absence of insoluble components was examined by visually comparing each sample.

[0061] Comparative sample 3 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaves into a 200 ml beaker and adding 95 g of olive oil to make a 5 w / w% solution.

[0062] Comparative sample 4 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaves into a 200 ml beaker, adding 1 g of emulsifier, and then adding 94 g of olive oil to make a 5 w / w% solution.

[0063] Comparative sample 5 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker, adding 1 g of emulsifier and 0.2 g of lecithin, and then adding 93.8 g of olive oil to make a 5 w / w% solution.

[0064] Confirmation sample 6 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker. First, 0.015 g (0.075 mg as fucose) of mozuku seaweed extract was added and mixed. Then, 1 g of emulsifier was added and mixed, followed by 0.2 g of lecithin and mixed. Finally, 93.785 g of olive oil was added and thoroughly emulsified and stirred to obtain a 5 w / w% solution. Table 2 shows the results of the oil solubilization test. [Table 2]

[0065] As can be seen from the results of comparative sample 3 shown in sample No. 3 of Table 2, even when oil is added to bitter melon leaf extract, precipitation of insoluble components occurs.

[0066] Therefore, as shown in comparative sample 4 (Sample No. 4), we first attempted to make the bitter melon leaf extract oil-soluble by adding an emulsifier, but this resulted in the formation of insoluble components.

[0067] Next, as shown in comparative sample 5 (Sample No. 5), we attempted to make the bitter melon leaf extract oil-soluble by adding an emulsifier and lecithin, but similarly, insoluble components were produced.

[0068] Next, as shown in confirmation sample 6 (Sample No. 6), when more mozuku water extract was added, no precipitation of insoluble components was observed.

[0069] These findings demonstrate that the method for solubilizing the ethanol extract of bitter melon leaves according to this embodiment is a method that can suppress the precipitation of insoluble components as much as possible.

[0070] [5. Examination of the precipitation-inhibiting effect of seaweed extracts other than mozuku] As described above, this application is characterized by the use of a water extract of mozuku seaweed as a precipitation inhibitor for the bitter melon leaf ethanol extract. Therefore, in this section, we investigated whether seaweed extracts other than mozuku seaweed can exert a precipitation inhibitory effect (suppression of precipitation) on the bitter melon leaf ethanol extract.

[0071] Since seaweed is classified into brown algae, red algae, and green algae, in this study, three types of seaweed water extracts were prepared using three types of seaweed: Fucus vesiculosus (brown algae), Utricularia japonica (red algae), and Ulva lactuca (green algae). The following confirmation tests were conducted to verify the precipitation prevention effect against the bitter melon leaf ethanol extract.

[0072] (5-1) Sample preparation The preparation of the bitter melon leaf ethanol extract was the same as described in [1. Preparation of bitter melon leaf ethanol extract] above. The seaweed water extract was prepared by first placing 100g of seaweed into a 1L beaker, adding 500g of water, and heating at 90°C for 3 hours while stirring well. Afterward, the seaweed was removed, and the resulting solution was used as the seaweed water extract. Three types of seaweed water extracts were prepared: Fucus vesiculosus as a brown algae water extract, Uriaria tetragonioides as a red algae water extract, and Ulva lactuca as a green algae water extract.

[0073] (5-2) Water solubility test Next, using the ethanol extract of bitter melon leaves mentioned above, we conducted a water solubility test to confirm whether the seaweed extracts of Fucus vesiculosus, Ulva lactuca, and Ulva lactuca produced similar effects to the water extract of Mozuku seaweed. Specifically, we investigated how the precipitation of insoluble components changed when water was added to the ethanol extract of bitter melon leaves, depending on whether or not each seaweed water extract was added.

[0074] The test involved visually comparing two samples: Comparative Sample 1, which was water-soluble without the addition of seaweed water extract, and Confirmation Samples 7, 8, and 9, which were water-soluble with the addition of seaweed water extract, to examine the presence or absence of insoluble components.

[0075] Comparative sample 1 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker and adding 95 g of water to make a 5 w / w% aqueous solution.

[0076] Furthermore, for confirmation samples 7, 8, and 9, 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaves was dispensed into a 200 ml beaker, 0.015 g of seaweed water extract was added, and then 94.985 g of water was added to make a 5% aqueous solution. Table 3 shows the results of the water solubility test. [Table 3]

[0077] As can be seen from the results of comparative sample 1, shown as sample No. 1, when water is added to the ethanol extract of bitter melon leaves, insoluble components precipitate. Similarly, the results of confirmation samples 7, 8, and 9 also confirmed the precipitation of insoluble components.

[0078] These findings indicate that the anti-precipitation effect on the bitter melon leaf ethanol extract is not an effect typically found in water extracts of common brown, red, or green algae, but rather a unique effect of the mozuku water extract.

[0079] (5-3) Oil-solubilization test Next, using the ethanol extract of bitter melon leaves mentioned above, we investigated whether the seaweed extracts of Fucus vesiculosus, Ulva lactuca, and Ulva lactuca produced the same effect as the water extract of Mozuku seaweed during oil solubility.

[0080] Specifically, we investigated how the precipitation of insoluble components changed when oil was added to an ethanol extract of bitter melon leaves, after adding an emulsifier and lecithin, depending on whether or not each seaweed water extract was added.

[0081] The test involved visually comparing comparative sample 5, in which an emulsifier and lecithin were added but each seaweed extract was not, with confirmation samples 10, 11, and 12, in which the emulsifier, lecithin, and each seaweed extract were added, to examine the presence or absence of insoluble components.

[0082] For confirmation samples 10, 11, and 12, 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf was dispensed into a 200 ml beaker. First, 0.015 g of seaweed water extract was added and mixed, then 1 g of emulsifier was added and mixed, followed by 0.2 g of lecithin and mixed. Finally, 93.785 g of olive oil was added and thoroughly emulsified and stirred to obtain a 5 w / w% solution. Table 4 shows the results of the oil solubilization test. [Table 4]

[0083] As can be seen from the results of comparative sample 5, shown as sample No. 5, when oil is added to the ethanol extract of bitter melon leaves, insoluble components precipitate, even in the presence of emulsifiers and lecithin. Similarly, the results of confirmation samples 10, 11, and 12 also confirmed the precipitation of insoluble components.

[0084] These findings indicate that the anti-precipitation effect on the bitter melon leaf ethanol extract is not an effect typically found in water extracts of common brown, red, or green algae, but rather a unique effect of the mozuku water extract.

[0085] [6. Confirmation test to determine if momordicin is involved in the precipitation of insoluble components] Regarding the dilution method of the bitter melon leaf ethanol extract, the oil-solubilizing method of the bitter melon leaf ethanol extract, and the use of mozuku water extract as a precipitation inhibitor for the bitter melon leaf ethanol extract according to this embodiment, the amount of mozuku water extract added to the bitter melon leaf ethanol extract can be such that, as mentioned above, the amount of fucose contained in the mozuku water extract is 0.01 to 1 mg per 1 mg of momordicin I contained in the bitter melon leaf ethanol extract.

[0086] Furthermore, as mentioned earlier, the inventors believe that momordicin is not directly involved in precipitation formation during dilution or oil solubility, nor is fucose directly involved in suppressing precipitation formation.

[0087] Therefore, in this section, we investigated whether momordicin is directly involved in precipitation during dilution or oil-solubilization by replacing the ethanol extract of bitter melon leaf with pure momordicin and then diluting it with water or oil.

[0088] Ideally, to investigate the involvement of momordicin, instead of using a standard bitter melon leaf ethanol extract, a solution from which only momordicin has been removed should be added to demonstrate that the precipitation of insoluble components is suppressed by the bitter melon leaf ethanol extract even in the absence of momordicin. However, removing only momordicin from the bitter melon leaf ethanol extract is extremely difficult.

[0089] Therefore, in this section, we used pure momordicin and set the final concentration of momordicin after water dilution or oil solubilization to twice the final concentration of momordicin derived from bitter melon leaf ethanol extract. We then investigated whether the amount of insoluble components precipitated when no water extract of mozuku seaweed was used changed, for example, whether it increased by about two times, in order to examine whether momordicin is directly involved in precipitate formation.

[0090] (6-1) Water solubility test Comparative sample 1 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker and adding 95 g of water to make a 5 w / w% aqueous solution. The final concentration of momordicin I in comparative sample 1 was 0.15 mg / 100 g.

[0091] Comparative sample 13 was prepared by adding 0.15 mg of momordicin I (purity 98% or higher, ChemFaces) to 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaves and dissolving it. This was then dispensed into a 200 ml beaker, and 95 g of water was added to make a 5 w / w% aqueous solution. The final concentration of momordicin I in comparative sample 13 was approximately 0.30 mg / 100 g. Table 5 shows the results of the water solubility test. [Table 5]

[0092] As shown in Table 5, precipitation of insoluble components was detected in both comparative sample 1 and comparative sample 13. Furthermore, the amount of precipitation in comparative sample 13 was almost the same as the amount precipitated in comparative sample 1, as visually compared.

[0093] These findings suggest that, in the dilution method of the bitter melon leaf ethanol extract according to this embodiment, and in the use of mozuku water extract as a precipitation inhibitor for the bitter melon leaf ethanol extract, momordicin is not directly involved in precipitation. It is very interesting that, despite momordicin not being directly involved in precipitation, the amount of momordicin can serve as a guideline in determining the amount of mozuku water extract to add to the bitter melon leaf ethanol extract.

[0094] (6-2) Oil-solubilization test Comparative sample 5 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker, then adding 1 g of emulsifier and mixing, followed by adding 0.2 g of lecithin and mixing, and finally adding 93.8 g of olive oil and thoroughly emulsifying and stirring to obtain a 5 w / w% solution. The final concentration of momordicin I in comparative sample 5 was 0.15 mg / 100 g.

[0095] Confirmation sample 14 was prepared by adding 0.15 mg of momordicin I (purity 98% or higher, ChemFaces) to 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaves and dissolving it. This was then dispensed into a 200 ml beaker, followed by the addition of 1 g of emulsifier and mixing, then 0.2 g of lecithin and mixing, and finally 93.8 g of olive oil and thoroughly emulsifying and stirring to obtain a 5 w / w% solution. The final concentration of momordicin I in comparison sample 14 was approximately 0.30 mg / 100 g. Table 6 shows the results of the oil solubilization test. [Table 6]

[0096] As shown in Table 6, precipitation of insoluble components was detected in both comparative sample 5 and comparative sample 14. Furthermore, the amount of precipitation in comparative sample 14 was almost the same as the amount precipitated in comparative sample 5, as visually compared.

[0097] These findings suggest that, in the oil-solubilizing method for the bitter melon leaf ethanol extract according to this embodiment, and in the use of mozuku water extract as a precipitation inhibitor for the bitter melon leaf ethanol extract, momordicin is not directly involved in precipitation.

[0098] [7. Confirmation test to determine if fucose is involved in the precipitation of insoluble components] Similar to the aforementioned [6. Confirmation test to determine whether momordicin is involved in the precipitation of insoluble components], this section investigates whether fucose is directly involved in precipitation during dilution or oil solubilization by replacing the water extract of mozuku with pure fucose and diluting it with water or oil.

[0099] Ideally, to investigate the involvement of fucose, it would be best to replace the regular mozuku water extract with a solution from which only fucose has been removed, and demonstrate that the precipitation of insoluble components is suppressed by the mozuku water extract even in the absence of fucose. However, removing only fucose from the mozuku water extract is extremely difficult.

[0100] Therefore, in this section, we decided to use pure fucose and investigate whether the precipitation of insoluble components is suppressed even when using an aqueous solution with the same fucose concentration as the water extract of mozuku seaweed, and to examine whether fucose is directly involved in precipitate formation.

[0101] (7-1) Water solubility test Comparative sample 1 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker and adding 95 g of water to make a 5 w / w% aqueous solution. The final concentration of momordicin I in comparative sample 1 was 0.15 mg / 100 g, and the final concentration of fucose was 0 mg / 100 g.

[0102] Comparative sample 15 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker, and adding 95 g of water containing 0.075 mg of fucose (purity 97.0% or higher, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) to make a 5 w / w% aqueous solution. The final concentration of momordicin I in comparative sample 15 was approximately 0.15 mg / 100 g, and the final concentration of fucose was 0.075 mg / 100 g. Table 7 shows the results of the water solubility test. [Table 7]

[0103] As shown in Table 7, precipitation of insoluble components was detected in both comparative sample 1 and comparative sample 15. This suggests that fucose is not directly involved in inhibiting precipitation when diluting the bitter melon leaf ethanol extract according to this embodiment or when using mozuku water extract as a precipitation inhibitor for the bitter melon leaf ethanol extract. It is very interesting that, despite not being directly involved in inhibiting precipitation, the amount of fucose can serve as a guideline when determining the amount of mozuku water extract to add to the bitter melon leaf ethanol extract.

[0104] (7-2) Oil-solubilization test Comparative sample 5 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker, then adding 1 g of emulsifier and mixing, followed by adding 0.2 g of lecithin and mixing, and finally adding 93.8 g of olive oil and thoroughly emulsifying and stirring to obtain a 5 w / w% solution. The final concentration of momordicin I in comparative sample 5 was 0.15 mg / 100 g, and the final concentration of fucose was 0 mg / 100 g.

[0105] Confirmation sample 16 was prepared by dispensing 5 g (0.15 mg as momordicin I) of ethanol extract of bitter melon leaf into a 200 ml beaker, adding 0.015 g of fucose water prepared by adding water to 0.075 mg of fucose, mixing, then adding 1 g of emulsifier and mixing, followed by adding 0.2 g of lecithin and mixing, and finally adding 93.785 g of olive oil and thoroughly emulsifying and stirring to obtain a 5 w / w% solution. The final concentration of momordicin I in comparison sample 16 was 0.15 mg / 100 g, and the final concentration of fucose was 0.075 mg / 100 g. Table 8 shows the results of the oil solubilization test. [Table 8]

[0106] As shown in Table 8, precipitation of insoluble components was detected in both comparative sample 5 and comparative sample 16. This suggests that fucose is not directly involved in suppressing precipitation in the oil-solubilizing method for the bitter melon leaf ethanol extract according to this embodiment, nor in the use of mozuku water extract as a precipitation inhibitor for the bitter melon leaf ethanol extract.

[0107] As described above, the method for diluting the bitter melon leaf ethanol extract according to this embodiment involves adding water to the bitter melon leaf ethanol extract to which a water extract of mozuku seaweed has been added, thus providing a method for diluting the bitter melon leaf ethanol extract with water that can suppress the formation of precipitate as much as possible.

[0108] Furthermore, according to the method for solubilizing the ethanol extract of bitter melon leaves according to this embodiment, an emulsifier and lecithin are added to the ethanol extract of bitter melon leaves to which a water extract of mozuku seaweed has been added, and then oil is added to prepare an emulsion, which is then added to the oil. This method provides an oil-soluble method for the ethanol extract of bitter melon leaves that can suppress the formation of precipitate as much as possible.

[0109] Finally, the above-described embodiments are merely examples of the present invention, and the present invention is not limited to the embodiments described above. Therefore, it goes without saying that various modifications can be made to embodiments other than those described above, as long as they do not depart from the technical spirit of the present invention, depending on the design and other factors.

Claims

1. A method for diluting a bitter melon leaf ethanol extract by adding water to a bitter melon leaf ethanol extract containing 1 part by weight of momordicin to which a bitter melon leaf water extract containing 0.0167 to 16.67 parts by weight of fucose has been added.

2. A method for producing a diluted solution of bitter melon leaf ethanol extract, comprising adding water to a bitter melon leaf ethanol extract containing 1 part by weight of momordicin to which a water extract of mozuku seaweed containing 0.0167 to 16.67 parts by weight of fucose has been added.

3. A method for oil-solubilizing a bitter melon leaf ethanol extract, comprising adding an emulsifier (provided that the final concentration after oil-solubilization contains lecithin equivalent to 0.1 to 50 mg / ml at the final concentration) to a bitter melon leaf ethanol extract containing 1 part by weight of momordicin, to which an aqueous extract of mozuku seaweed containing 0.0167 to 16.67 parts by weight of fucose has been added, and further adding oil to prepare an emulsion, and adding the emulsion to the oil.

4. A method for producing an oil-soluble solution of a bitter melon leaf ethanol extract, comprising adding an emulsifier (provided that the final concentration after oil solubilization contains lecithin equivalent to 0.1 to 50 mg / ml) to an ethanol extract of bitter melon leaves containing 1 part by weight of momordicin, to which an aqueous extract of mozuku seaweed containing 0.0167 to 16.67 parts by weight of fucose has been added, further adding oil to prepare an emulsion, and adding the emulsion to the oil.

5. Use of a water extract of mozuku containing 0.0167 to 16.67 parts by weight of fucose as a precipitation inhibitor for a bitter melon leaf ethanol extract containing 1 part by weight of momordicin.

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