KR100190989B1 - Water soluble fraction extracts of pinelliae tuber having improved skin whitening effects by removing darkening components and a composition for improvining freckles and skin whitening containing thereof - Google Patents

Abstract

The present invention relates to a semi-aqueous water-soluble fraction extract having a blackening effect removed from the halves and improving the whitening effect thereof, and a composition for improving blemishes, freckles and skin whitening containing the same.

Half of the water-soluble fraction extract of the present invention is half or more times extracted with lower alcohol, ethyl acetate or acetone having 1 to 4 carbon atoms, water, anhydrous or anhydrous, and the polar solvent fraction containing blackening component after concentration under reduced pressure And then purified and dried to obtain an aqueous fraction.

The composition according to the invention is characterized in that it contains 0.0001 to 10% by weight of a semi-soluble fraction extract in dry weight.

Description

Semi-water soluble fraction extract with improved whitening effect by removing blackening components and composition for improving blemishes, freckles and skin whitening containing same

The present invention relates to a water-soluble fraction extract and a composition containing the same, which is characterized in that the whitening effect is improved by removing the blackening component from the halves extract, and more particularly, the improved halves having excellent melanogenesis inhibitory effect and whitening effect. It relates to a water-soluble fraction of the extract and the composition containing the blemishes, freckles and skin whitening.

To have white and fair skin is everyone's constant desire. Human skin color is genetically determined by the concentration and distribution of melanin in the skin, but is also influenced by environmental or physiological conditions such as solar ultraviolet light, fatigue and stress. Melanin is a type of amino acid tyrosine (tyrosine) enzymes called tyrosinase (tyrosinase) acts to convert to dopa (dopa), dopaquinone (dopaquinone) is produced through a non-enzymatic oxidation reaction. However, although the pathway through which melanin is made is known, it is still unknown in detail the mechanism that induces melanin synthesis, which is a prior step of tyrosinase action.

Therefore, conventionally, a substance having an inhibitory activity on tyrosinase such as hydroquinone, ascorbec acid, kojic acid, glutathione, and the like is added to the ointment or cosmetics, and the skin whitening, blemishes, and freckles are used. Although it has been used for the purpose of improvement, the use is limited because it causes irritation to the skin or poor product stability, and the effect was also weak.

Therefore, the present inventors have developed a new whitening agent for animals and plants native to nature using mouse melanoma cells (B16 melanoma cells) that can be screened to a substance that inhibits melanin synthesis induction itself.

In Japanese Patent Laid-Open No. 6-219934, on the other hand, herbal extracts such as powders have been reported to inhibit the pigment biosynthesis of rat melanoma cells (b16 melanoma cells).

However, the half-harvest extracts quoted in Pyeong 6-219934, when used as they are, are very difficult to apply to actual products because they have a weak effect and have a dark blackish brown color. Therefore, the present inventors devote to research and found that the half extract has a whitening effect as well as a component that strongly blackens rat melanoma cell (b16 melaonma cell), so that the whitening effect of the whole extract is weak. In addition, the blackening component separated from the halved extract has been patented as a skin blackening agent (Korean Patent Application No. 95-34976).

Subsequently, when the blackening component mixed in a large amount is removed, the water-soluble fraction in the extract significantly improves the whitening effect, and the white precipitate produced by adjusting the water-soluble fraction to pH 6.0-13.0 with alkali is the main component showing the whitening effect. It was found that the present invention has been completed.

The present invention will be described in detail as follows.

Buy half of the medicinal herb and grind it finely, and crush it to 50 ~ 100 in an extractor equipped with reflux cooler with 5 ~ 20 volume of water or 1 ~ 4 carbon number anhydrous or hydrous lower alcohol, ethyl acetate or acetone. Extract by heating to 1 ~ 5 hours. After filtering with a filter cloth, the residue is extracted one more time in the same manner. The extracts were combined, concentrated under reduced pressure, filtered out and concentrated again. Then, purified water is added to the concentrated primary extract, and the fractions dissolved in a polar organic solvent having a polarity of 3.0 or more that is not mixed with water are purified and dried to obtain a semi-soluble fraction extract.

In addition, alkali is added to the purified semi-soluble fraction extract to obtain a white precipitate.

As a result of applying the purified product thus obtained to the rat melanoma cells (B16 melanoma cells) compared to the half-half total extract in Japanese Patent Laid-Open No. 6-219934, the half-half total extract has no whitening effect or is very weak. On the other hand, when the blackening component that promotes melanin synthesis is removed as in the present invention, the whitening effect is significantly more than 10 times, and the precipitate further purified in an alkaline state has excellent whiteness and the whitening component is concentrated, such as ointment or lotion. Even if manufactured, there was no problem in the color or formulation of the product, and even when applied to human skin, it was confirmed that it shows a safe and excellent whitening effect. However, the total extract, which does not remove the blackening component, was difficult to expect a whitening effect, rather irritating, and there was a problem in the formulation, such as blackish brown in the product.

The content of the semi-soluble water extract fraction of the present invention contained in skin external ointment or cosmetic preparation is suitable for the dry weight of 0.0001 ~ 10% by weight. A clear effect could not be expected at concentrations below 0.0001% by weight, and no increase in effect was seen at concentrations above 10% by weight.

Next, the present invention will be described in detail with reference to Examples, Comparative Examples, and Experimental Examples, and the present invention is not limited to Examples.

Comparative Example 1

The lower tuber pulverized product was put into 1.5 L of 80% methanol solution, boiled for 3 hours in a reflux extractor equipped with a cooling capacitor, filtered through a 300 mesh filter cloth, and the residue was extracted once more in the same manner. The extract was filtered out of whatman No. 2 filter paper at room temperature, concentrated under reduced pressure at 60 ° C. in a distillation apparatus equipped with a cooling condenser, and then spray-dried to obtain a dry weight of 12.1 g.

Comparative Example 2

120 ml of purified water was added to the first half extract obtained in Comparative Example 1, the same amount of normal hexane was added to the mixture, and the mixture was mixed well. Then, the normal hexane fraction was removed, and the same amount of methylene chloride was added to the filtrate. The process of obtaining was repeated three times and concentrated under reduced pressure at 40 ° C. in a distillation apparatus equipped with a cooling condenser to obtain a dry weight of 1.2 g.

Comparative Example 3

After adding the same amount of normal butanol to the filtrate obtained in Comparative Example 2 and mixing it well, the process of obtaining a fraction of normal butanol was repeated three times, and concentrated under reduced pressure at 50 ° C. in a distillation apparatus equipped with a cooling condenser to obtain a dry weight of 2.5 g.

Example 1

The filtrate (water-soluble fraction) obtained in Comparative Example 3 was concentrated under reduced pressure at 60 ° C. in a distillation apparatus with a cooling condenser to obtain a dry weight of 7.4 g.

[Comparative Example 4]

The lower tuber pulverized product 300g was performed at 50 degreeC by the method of Unexamined-Japanese-Patent No. 6-219934, and the dry weight of 10.5g was obtained.

[Comparative Example 5]

100 ml of purified water was added to 10.5 g of the halves extract obtained in Comparative Example 4, and the drying was performed in the same manner as in Comparative Example 2 to obtain a dry weight of 0.9 g.

Comparative Example 6

The filtrate in Comparative Example 5 was subjected to the same method as in Comparative 3 to obtain 2.1 g of a dry weight.

Example 2

The filtrate (aqueous fraction) in Comparative Example 6 was carried out in the same manner as in Example 1 to obtain a dry weight of 6.8 g.

Example 3

35 ml of purified water was added to the dried product of Example 1, the reaction mixture was titrated to pH 12.5 with sodium hydroxide and sodium solution, and the white precipitate obtained by centrifugation was concentrated under reduced pressure at 60 ° C. in a distillation apparatus equipped with a cooling condenser to obtain a dry weight of 0.5 g.

Experimental Example 1

Each of the extracts obtained in Examples 1 to 3 and Comparative Examples 1 to 6 were added to the culture solution of ear melanoma cells (B16 mouse melanoma cells) to test the whitening effect at the cellular level (Lotan r., Lotan d. Cancer Res. 40: 3345-3350, 1980). Each of the extracts obtained in Examples 1 to 3 and Comparative Examples 1 to 6 and hydroquinone were added to the culture medium of rat B16 melanoma cells, and then cultured for 3 days, followed by trypsin treatment. The melanin was extracted after centrifugation and removal from the vessel. To this, 1 ml of sodium hydroxide solution (1N concentration) was added, boiled for 10 minutes to dissolve melanin, and the absorbance was measured at 400 nanometers (nm) using a spectrophotometer to measure the amount of melanin produced per unit cell number (10 ⁶ cells) It was performed by the method shown as, the relative melanin production relative to the control was calculated as the inhibition rate (%) and the results are summarized in Table 1.

As can be seen from the above results, it can be seen that when compared to the control example of Comparative Examples 2 to 3, the solvent fraction of the lower extract has a blackening component that strongly promotes melanin production despite the application amount of the trace amount. However, it can be seen that the water-soluble fraction from which the blackening component is removed as in Example 1 according to the present invention increases the whitening effect by more than 10 times at the same concentration as in Comparative Example 1. Therefore, it can be seen that the weakening whitening effect of the total extract is a phenomenon in which blackening components that promote melanin production cancel the melanin production inhibitory effect. In addition, the whitening effect is remarkably improved in Example 2, in which blackening component of Comparative Examples 5 and 6 is present in Comparative Example 4 according to Japanese Patent Laid-Open No. 6-219934 and the blackening component is very weak. I can see

In addition, the whitening effect of Example 3, which further refines the water-soluble fraction of the lower half, is 13 times higher than that of Example 1, and can be applied in small amounts. As a white powder, it is understood that there is no problem in the color of the product at all. Can be.

Experimental Example 2

When the melanin production inhibitory ability of the rat melanoma cells cultured in the same manner as in Experimental Example 1 was investigated according to the concentration, the results are shown in Table 2.

As can be seen from the above results, the extract of Example 3 has a very strong melanin production inhibitory ability against cultured mouse melanoma cells, and melanin production inhibitory ability from 0.0001% to 0.1% or more depending on the concentration of the extract of Example 3 This increases. In addition, hydroquinone is excellent in whitening effect, but can not be experimented at 0.0001% or more due to cytotoxicity such as yesterday's cell growth and cell type change, extract of Example 3 is cytotoxic even at 0.1% concentration It has higher melanin production inhibition than hydroquinone without showing.

The preparation example of the skin whitening composition containing the extract of the Example below is shown by the manufacture example.

[Production Example 1]

An example of the external preparation of the ointment for the skin containing the half-harvest extract is as follows.

Herein, the semi-half extract refers to that of Example 1.

Production Example 2 and Comparative Production Example 1

A prescription example of a cream in cosmetics containing a half herb extract is as follows.

Here, the half extract refers to that of Example 3.

Production Example 3 and Comparative Production Example 2

Prescription examples of the flexible cosmetics in the cosmetics containing the semi-half extract are as follows.

Here, the half extract refers to that of Example 3.

Production Example 4 and Comparative Production Example 3

An example of the prescription of nutrient cosmetics in cosmetics containing half the extract is as follows.

Here, the half extract refers to that of Example 3.

Production Example 5 and Comparative Production Example 4

A prescription example of the pack among the cosmetics containing half the extract is as follows.

Here, the half extract is that of Example 3.

Production Example 6 and Comparative Production Example 5

An example of the prescription of the essence among the cosmetics containing half the extract is as follows.

Here, the half extract refers to that of Example 3.

[Comparative Production Example 6 and Comparative Production Example 7]

A prescription example of a cream in cosmetics containing a half herb extract is as follows.

Here, the half bottom extract says the thing of the comparative example 4.

[Comparative Production Example 8 and Comparative Production Example 9]

The formulation of the cream in the cosmetics containing half-harvest extract is as follows.

Here, the half bottom extract says the thing of the comparative example 4.

Experimental Example 3

20 healthy males and females were placed in the lower arm of each arm with 6 holes of 7 mm diameter in two rows of aluminum foil, and 60mJ / cm ² light was irradiated with a ORIEL solar simulator 1000w at a distance of 10 cm from the arm. . The irradiation site was washed well with 70% ethanol aqueous solution before irradiation. Sample containing semi-extracts prepared according to Preparation Examples 2,3,6, and Comparative Preparation Examples 1,2,5,6,7,8,9, twice a day from 3 days before irradiation to 3 weeks after irradiation. Base and half-free extracts were applied to the same row in pairs. For each, the pigmentation degree of the production example and the comparative production example were visually determined, and the evaluation of the degree of suppression of the pigmentation of the production example compared to the comparative production example was evaluated in three stages of distinct effects, effects, and no differences. Table 3 is as follows.

As can be seen from the results in Table 3 above, the half water-soluble fraction extract-containing cosmetics prepared according to Preparation Examples 2, 3 and 6, prepared using Example 3, were used for at least 10 of 20 subjects. It showed a whitening effect and did not show any side effects, indicating that the improved water-soluble fraction extract of the improved half was safe and very effective in improving freckles or freckles or skin whitening agents. In addition, there was no problem in the color or formulation of the product using a purified sample in the product. However, Comparative Preparation Example 6 prepared using Comparative Example 4 does not exhibit the same remarkable whitening effect as Preparation Example 2, because the whitening effect is canceled by the blackening component contained in the lower half. In addition, even if the concentration was increased 10 times (1% content) as in Comparative Preparation Example 8, the whitening effect was not recognized. Particularly, the product was dark brown, and there was a problem with the formulation, and some of the irritants were appealed. However, in Example 3, Preparation Examples 2, 3, and 6 do not show any side effects, it can be seen that the water-soluble fraction extract of the contrary is a safe and highly effective freckles or freckles improvement or skin lightening agent.

Claims (3)

Extracting half or more times with water or anhydrous lower alcohol, ethyl acetate or acetone having 1 to 4 carbon atoms and concentrating the extract under reduced pressure; Adding purified water to the concentrate and adding a polar organic solvent having a polarity of 3.0 or more that is not mixed with water to perform fractionation to remove the organic solvent fraction; Aqueous fraction extract of Banja extract, characterized in that it is prepared by drying the water-soluble fraction from which the organic solvent fraction is removed. The method of claim 1, wherein the water-soluble fraction extract of the half organic extract is prepared by a method of obtaining a precipitate by further adjusting the water-soluble fraction extract from the polar organic solvent fraction to pH 6.0 ~ 13.0 with an alkali. The composition for improving blemishes, freckles and skin whitening, characterized in that it contains 0.0001 to 10% by weight of the extract of claim 1 or 2 as a dry weight.