KR100971286B1 - Method for repressing cytotoxicity of chinese medicine with lightening effect for skin - Google Patents

Abstract

The present invention relates to a method for reducing the cytotoxicity of herbal ingredients with skin whitening ability by fermenting the herbal extract yogurt, the herbal skin cosmetic pack developed in the present invention is characterized by showing excellent skin whitening ability while irritating or toxic have.

Toxic, whitening, cells, yogurt, pack, skin, herbal

Description

Method for reducing cytotoxicity of herbal ingredients with skin whitening ability {Method for repressing cytotoxicity of chinese medicine with lightening effect for skin}

The present invention relates to a method for reducing the cytotoxicity of herbal ingredients with skin whitening ability, and more particularly, to a method for reducing the cytotoxicity of herbal ingredients with skin whitening ability by fermenting herbal extracts.

After the IMF, despite the overall downturn in the domestic economy, the cosmetics industry has exploded and the current market size reaches 5 trillion won. In particular, due to changes in the economic flows caused by the IMF and the aging measures of old age populations due to life extension, the age of the consumer population has risen from 10 to 30 to 40 to 60.They hate not looking old and not wrinkled. In addition to cosmetics, functional cosmetics having a specific function is also very preferred.

Herbal cosmetics are currently on the market so that most domestic cosmetic companies have more than one brand. The representative one is Sulwhasoo, which was launched in the Pacific in 1997, and its 2004 sales were 330 billion won.

In addition, Chinese clay cosmetics are gaining popularity as Chamtowon's ocher packs generate sales of KRW 32 billion and Able C & C's herbal cosmetics 'Mishaok Yongsan' exceeded one million sales in six months of its release. Accordingly, the market of oriental cosmetics has grown day by day, and the market size of this year is estimated to be about 70 billion to 800 billion won.

The herbal pack has a big impact on the growth of such cosmetics. Traditionally, there are many people who have made and used the necessary packs from consent and knowledge gained from folk remedies, and herbal packs are not rejected by consumers, and the effects are known to the general public and are easier to enter the market than other cosmetic products. Could.

Meanwhile, the human skin color is determined by the amount of melanin, carotene, and hemoglobin, of which melanin plays the most important role. Melanin is produced through the enzymatic and non-enzymatic oxidation of tyrosine in cells called melanocytes in the basal layer of the skin epidermis and is transferred to the keratinocytes constituting the epidermis.

Melanin pigment produced by the melanocytes of the skin of the body is a phenolic polymer material having a complex form of a black pigment and protein, and plays an important role in inhibiting skin damage by ultraviolet radiation from the sun.

However, excessive synthesis and accumulation of melanin not only causes serious aesthetic problems such as blemishes, freckles and senile surpluses on the skin, but also promotes skin aging and causes skin cancer.

It is known that the action of tyrosinase in melanocytes is the most important for melanin biosynthesis. Most studies focused on the inhibition of tyrosinase activity by preventing melanin overproduction and skin whitening and skin cancer. have. Many compounds with tyrosinase inhibitory activity have already been used as raw materials for whitening cosmetics, but their use is restricted due to poor stability due to poor decomposition, odor, unclear efficacy and safety issues at the biological level. to be.

Therefore, in recent years, in order to prevent and improve such a problem, relatively low toxicity, and search for a herbal medicine having excellent whitening action, and the technology development using this has been performed. As an example, Korean Patent Publication No. 1993-0010548 (whitening cosmetics containing the extract of a white bark), Korean Patent Publication No. 10-0535875 (cosmetic composition containing a plant mixture extract having a skin whitening effect), Korean Registered Patent Publication No. 10-0654959 (a cosmetic composition for whitening) and the like have been published in the patent literature, but the use of herbal medicines has caused problems of skin irritation and toxicity and has been problematic for application to the skin at high concentrations, and has been used at lower concentrations. In this case, there has been a problem that the whitening ability is not properly exercised.

Accordingly, an object of the present invention is to develop and provide a method for reducing the cytotoxicity of herbal materials having skin whitening ability.

In order to achieve the above object, the present invention provides a first embodiment of the present invention, 0.3% (w / v) for Angelica, Cheonggung, Sukjihwang, Bokryeong, and Baekchul, and 0.25% (w / v) for Jujube and Cinnamon, respectively. And, yulmu, yulpi, mung beans, passersby each 0.5% (w / v) put in water to extract the hot water to obtain a Chinese medicine hydrothermal extract (a); Diluting the obtained herbal extract with milk to 1/4, and adding 5% (w / v) skim milk powder and 2% (w / v) sugar and fermenting to prepare a yoghurt (b); And, (c) packing the prepared yogurt; It provides a method for reducing the cytotoxicity of the herbal material with skin whitening ability comprising a.

In addition, the present invention is a second aspect of the present invention, 4% (w / v) each of mung bean, white paper, white weight, baekryeom, baekjaja, baekbuja, cheonhwa powder, 2% (w / v) persimmon fragrance, samraeja, kwakyang / v) and each of the Youngyang Hyangjeop, windproof, honbon, flakes in water by 0.8% (w / v) each to obtain a Chinese medicine hydrothermal extract by hydrothermal extraction (a); Diluting the obtained herbal extract with milk to 1/4, and adding 5% (w / v) skim milk powder and 2% (w / v) sugar and fermenting to prepare a yoghurt (b); And, (c) packing the prepared yogurt; It provides a method for reducing the cytotoxicity of the herbal material with skin whitening ability comprising a.

Hereinafter, the problem solving means of the present invention will be described in more detail.

Skin whitening is the desire of all people, both young and old, and efforts to discover new materials with skin whitening ability in recent years have been made targeting a variety of sources. Recently, herbal materials are rapidly emerging as the target of discovering new skin whitening materials in line with the spread of well-being culture and new awareness of traditional Chinese medicine.

By the way, the herbal material has a high cytotoxicity when used at high concentrations causes undesirable problems, and when used at low concentrations, effective skin whitening ability can not be expected.

Therefore, in view of the use of herbal materials, it would be required to develop a new method of reducing the cytotoxicity while showing skin whitening ability, the inventors of the present invention as a result of the yogurt fermentation of herbal ingredients with skin whitening ability, The present invention has been completed by discovering that the cytotoxicity is significantly reduced.

On the other hand, even if the cytotoxicity is reduced, this is a big problem if the original purpose of skin whitening ability can not be achieved, the herbal pack prepared by the method of the present invention, while the cytotoxicity is reduced, the skin whitening ability is rather improved. do. This is believed to have occurred because the herbal skin cosmetic pack prepared by the method of the present invention increased the skin penetration of the active ingredient derived from the herbal medicine compared to the developed herbal medicine extract as a simple pack.

In general, when the acidity is lowered to pH 5 or less, plant-derived glycosides in the extract are hydrolyzed in the form of aglycone. Aglycone-type compounds have a lower molecular weight than glycosides and are easier to penetrate into the skin than glycosides. Yoghurt is a milk-based beverage that is fermented with lactic acid bacteria and has a pH of around 4 . In yogurt fermentation, it could be concluded that the useful ingredients derived from glycosides were decomposed into aglycone and the skin permeability was enhanced by this change in pH. It confirmed through Experimental Example 2.

As described above, the method for reducing the cytotoxicity of the herbal material having skin whitening ability according to the present invention provides a method for preparing a new herbal pack which can show excellent skin whitening ability while stimulating or toxicity is alleviated.

Hereinafter, the content of the present invention will be described in detail through the following examples, but the scope of the present invention is not limited only to the following examples, and includes modifications of equivalent technical ideas.

Example 1: Preparation of Herbal Yogurt Pack

To make yoghurt pack (OK1), add Angelica, Cheonung, Sukjihwang, Bokryeong, Baekchul, 0.3% each, Jujube, Cinnamon 0.25% each, and Yulmu, Yulpi, Mung Bean, Almonds, 0.5% each in 1 liter of water Extracted.

Then, the herbal extracts obtained by hot water extraction were diluted with milk to 1/2, 1/4, 1/8 and 1/16, respectively, and then 5% (w / v) skim milk powder and 2% (w / v) sugar. ) And yogurt fermentation to prepare a herbal yogurt pack.

The yogurt prepared by the above process was sealed in a packaging pack commonly used in the cosmetic industry to prepare a skin massage pack.

Example 2: Preparation of Herbal Yogurt Pack

4% (w / v) of mung bean, white paper, white weight, baekrye, baekpjab, baekbuja, and hwahwa each used as a pack to make yogurt pack (OK3), and 2% of each / v), and Young Hyang, Windproof, Gobon, and Sculptor were each extracted with hydrothermal extract in 1 liter of 0.8% (w / v).

Then, the herbal extracts obtained by hot water extraction were diluted with milk to 1/2, 1/4, 1/8, 1/16, respectively, and then 5% (w / v) skim milk powder and 2% (w / v) sugar. ) And yogurt fermentation to prepare the herbal yogurt pack of the present invention.

The herbal yogurt prepared by the above process was sealed in a packaging pack commonly used in the cosmetic industry to prepare a skin massage pack.

Experimental Example  1: pH measurement of herbal yogurt

The pH of the herbal yoghurt packs prepared in Example 1 (OK1 with herbal extracts diluted in quarters) was analyzed and found to be slightly acidic as pH 4.2. This means that the acidity is lowered through the fermentation and ripening process. In view of the fact that when the acidity is lowered below pH 5, glycosides present in the plant are decomposed into aglycone form, according to this weak acidification, herbal medicines added to herbal yogurt may be decomposed into aglycone and become low molecular weight. It can be inferred that this may ultimately lead to an increase in the absorption capacity of the herbal medicine active ingredient into the skin.

On the other hand, if the pack product is acidic, the exfoliation is naturally removed by the acid, and the environment of the skin is improved. The removal of the stratum corneum promotes the absorption of nutrients that affect the skin. Thus, the skin absorption ability can be further promoted.

As such, the pH of the pack is very important in the absorption capacity of the active ingredient, when yogurt fermentation is carried out as in the present invention, the pH can be naturally lowered to weak acidity without the addition of artificial pH adjusting agent.

Experimental Example  2: of herbal medicine active ingredient Low molecular weight  black

Microorganisms grow by decomposing organic matters, and thus, microbial growth can be thought of as decomposition of natural substances, which means that the decomposition of these natural substances means that the high molecular compound is changed to low molecules.

Lactic acid bacteria break down sugars such as glucose to produce two molecules of lactic acid, which results in an acidic pH. The decomposition of hexasaccharides such as glucose into two molecules can be indirectly assayed as a change in pH.

In this experiment, by inoculating and cultivating the lactic acid bacteria in the herbal extract obtained in the intermediate step of the herbal yogurt preparation step of Example 1 to prove that the lactic acid bacteria are indirectly to prove the low molecular weight of natural products.

500 ml of herbal extracts were placed in a sterile fermenter and inoculated with 20 ml of lactic acid bacteria that were precultured. Fermentation samples were taken every 24 hours while the fermentation was carried out at an impeller speed of 800 rpm under the conditions of 30 ° C to measure the acidity. After 4 days of fermentation, the same operation was repeated by adding 300 ml of the herbal extract again.

The results were as shown in Figure 1, it was confirmed that the pH drops to around 3.3 as the fermentation proceeds in the vicinity of the initial 5.9.

On the other hand, to examine the amount of organic acid and total acid by lactic acid bacteria culture, 500ml of herbal extracts were incubated for 6 hours or 12 hours according to the above method. The culture solution was recovered, centrifuged to remove solids, and the supernatant was transferred to a clean container. The supernatant was diluted three times with distilled water and then sterile filtered with a 0.2 μm membrane filter. The filtered sample was used for HPLC analysis.

HPLC analysis was performed using Shimazu Model CBM-10A (Japan) according to the following conditions. The column used 'Supelcogelc-610 ((30 cm x 7.8 mm ID) "and the mobile phase consisted of 0.1% H ₃ PO ₄ at a rate of 0.6 mL / min.

Injection volume was 10 μl, and the test temperature was maintained at 30 ° C. Citric acid, tartaric acid, malic acid, succinic acid, lactic acid, acetic acid, and formic acid were used as standards. Was as Table 1 below.

division 0 hours 6 hours after fermentation 12 hours after fermentation Citric acid 4.53 4.92 4.73 Tartaric acid - - - Malian 181.51 214.16 136.63 Suche mountain - - - Lactic acid 878.37 1435.37 2068.38 Formic acid - - - Acetic acid 7.66 5.22 1.41

Tartaric acid, succinic acid, and formic acid were not detected, and citric acid and malic acid were increased during the culturing process and decreased during the 12 hour culture. And it showed a tendency to decrease gradually with acetic acid culture. Since lactic acid bacteria were inoculated, lactic acid increased continuously with culture.

As a result of measuring the total acid, it was found that the fermentation was 12 times higher than the herbal extract. This means that by inoculating and fermenting lactic acid bacteria, the sugars in the medium are broken down so that the lactic acid bacteria are multiplied to increase the amount of total acid.

From the above results, it would be indirectly proved that the sugar component in the medium was degraded and lowered due to the change of pH and total acid according to the lactic acid bacteria culture.

Experimental Example  3: Skin whitening  Synergy test

In this experiment, the inhibitory activity of tyrosinase of the herbal yogurt pack prepared in Example was investigated.

Herbal yogurt packs prepared in Examples 1 (OK1 containing herbal extracts diluted in 1/4) and Example 2 (OK3 containing herbal extracts diluted in 1/4) were used as the experimental group, and the control group was used as a control. After diluting each herbal extract (Comparative Example 1 (OK5) and Comparative Example 2 (OK6), respectively) obtained in the intermediate stages of the herbal yogurt pack preparation of Examples 1 and 2 by 1/4 with milk. The sample was used without further treatment.

For the experiment, 50 μl of sample and 50 μl of 10 U / ml tyrosinase were mixed in a 96 well ELISA plate, and then 100 μl of substrate mix was added and reacted at 37 ° C. for 20 minutes. Tyrosinase activity was measured as absorbance at 475 nm and then expressed as a relative value to absorbance of the untreated group (UT). Statistical significance of the sample treatment group for UT was indicated by * (p <0.05), ** (p <0.01) or *** (p <0.001). All data were tripled experiments.

The results were the same as in Figure 2, compared with the group not treated with herbal biopack OK1, including the herbal extracts diluted 1/4, OK3 containing the herbs diluted 1/4 (78.5%) significant inhibition effect However, 95.6% of OK5 diluted with 1/4 of herbal extracts with milk and 9% of OK6 diluted with 1/4 of herbal extracts with milk were 90.7%. It is not shown (Fig. 2).

This means that the usefulness of the medicinal herb extract can be increased by fermentation of the medicinal herb extract into yogurt, and various glycosides contained in the medicinal herb extract are hydrolyzed into various kinds of aglycone to inhibit tyrosinase of the whole yogurt composition. It can also be interpreted that this has been increased.

Experimental Example  4: Cytotoxicity Assay of Herbal Yogurt Packs

In this experimental example, the cytotoxicity of the herbal yogurt pack of the present invention was investigated. Herbal yogurt packs prepared in Examples 1 (OK1 containing herbal extracts diluted in 1/4) and Example 2 (OK3 containing herbal extracts diluted in 1/4) were used as the experimental group, and the control group was used as a control. After diluting each herbal extract (Comparative Example 1 (OK5) and Comparative Example 2 (OK6), respectively) obtained in the intermediate stages of the herbal yogurt pack preparation of Examples 1 and 2 by 1/4 with milk. The sample was used without further treatment.

The results were as shown in Table 2 below, but showed less cytotoxicity of the yogurt pack.

Mena ON at 490nm Cytotoxicity (%) No treatment group 0.000 0.0% OK1 -0.140 -4.2 OK3 -0.145 -4.4 OK5 0.103 3.1% OK6 0.227 6.9% + 3.306 100.0%

1 is a graph showing the pH change of the herbal medicine extract according to the lactic acid bacteria culture.

2 is a graph showing the inhibitory effect of tyrosinase activity of samples.

Claims (2)

0.3% (w / v) for Angelica, Cheonggeung, Sukjihwang, Bokryeong, and Baekchul, 0.25% (w / v) for Jujube and Cinnamon, respectively, and 0.5% (w / v) for Yulmu, Yulpi, Mung Bean, and Pedestrian ) Into each water to extract hot water to obtain a Chinese medicine hot water extract (a); Diluting the obtained herbal extract with milk to 1/4, and adding 5% (w / v) skim milk powder and 2% (w / v) sugar and fermenting to prepare a yoghurt (b); And, Step (c) of packaging the prepared yogurt; Method of reducing the cytotoxicity of the herbal material with skin whitening ability comprising a; 4% (w / v) of mung bean, white paper, white weight, baekryeom, baekjaja, baekbuja and cheonhwa, respectively, 2% (w / v) of persimmon, Samraeja, and Gwakyang, respectively. Step (a) to obtain the Chinese herbal hydrothermal extract by putting the engraver into water each 0.8% (w / v) by hydrothermal extraction; Diluting the obtained herbal extract with milk to 1/4, and adding 5% (w / v) skim milk powder and 2% (w / v) sugar and fermenting to prepare a yoghurt (b); And, Step (c) of packaging the prepared yogurt; Method of reducing the cytotoxicity of the herbal material with skin whitening ability comprising a;

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