KR20120017354A - Cosmetic composition for skin whitening comprising red rice extract - Google Patents

Abstract

PURPOSE: A cosmetic composition containing red rice extract for skin whitening is provided to suppress tyrosinase activity and to block melanogenesis with less cytotoxicity. CONSTITUTION: A cosmetic composition for skin whitening contains 0.001-20 wt% of red rice extract. The extract is prepared from rice bran of red rice using distilled water, 1,3-butylene glycol, alcohol of 1-4 carbon atoms, ethyl acetate, or acetone solvent. The cosmetic composition is used in the form of an external use skin ointment, essence, lotion, cream, gel, lotion, pack, massage cream, emulsion, foundation, makeup base, soap, or sun oil.

Description

Cosmetic composition for skin whitening containing red rice extract

The present invention relates to a cosmetic composition for skin whitening containing a red rice extract. The red rice extract of the present invention is very excellent in whitening activity as compared to the white rice can be usefully used as a cosmetic composition for skin whitening.

Recently, the tension of skin cells such as UV exposure and drying due to many outdoor activities such as golf, mountain climbing, hiking, fishing, and so on, is a direct cause of skin aging. Currently, functional cosmetics are focused on UV protection and whitening function, and thus, it is necessary to rejuvenate the skin through the relaxation of the underlying skin nerve and cell recovery function.

Conventionally, arbutin has been mainly used as an active ingredient of a cosmetic composition having a whitening activity, but the compound has a problem of causing many side effects when the compound is not used in a large amount.

Therefore, as a component having a whitening activity of the cosmetic composition has become a lot of attention to natural resources, among which many cosmetic compositions using rice extract has been developed. In particular, the rice used as the cosmetic composition is mostly white or black rice, the cosmetic composition using the white or black rice extract was mainly developed. However, in general, various products using white rice and black rice have been developed, but there are no products using red rice by wild type or artificial breeding and natural mutation.

The inventors of the present invention while studying a method of using the red rice as a material of the cosmetic composition, the red rice extract is superior to the white or brown rice extract, the whitening activity is excellent, and the cell stability is also very excellent as a cosmetic composition for whitening The present invention has been found to be useful.

Therefore, an object of the present invention is to provide a cosmetic composition for skin whitening containing red rice extract.

In order to solve the above problems, the present invention provides a cosmetic composition for skin whitening containing a red rice extract.

Hereinafter, the present invention will be described in more detail.

In the present invention, the red rice is a rice having red color on the skin or seedling, but is not limited thereto. Preferably, red pearl wine (National Species Variety Registration No. 1-1-2001-5) or red pearl wine (National Species Resource Variety No. 1- 1-2007-7) may be rice harvested from varieties of rice. More preferably it may be rice harvested from the rice of red pearl variety. The red rice is scientific name Oryza sativa L. and the genotype of the rind may be Rd and RC. The red rice RC gene is a gene encoding a basic Helix-Loop-Helix protein, but is not limited thereto. Preferably, the NCBI registration number may be AB247503 or AB250059, and the Rd gene encodes dihydroflavonol-4-reductase (DFR). Although not limited thereto, the NCBI registration number may be AP004317. In particular, rice may be divided into white, brown, red, purple, or black and green rice according to the genotype of the skin, that is, the expression level of the pigment is changed depending on the combination of genes involved in the skin formation. It may be represented by red or the like.

In the present invention, the rice bran is preferably composed of shell of brown rice, that is, bran layer (peel, seed skin, whistle layer) and rice snow, which are easily obtained by those skilled in the art in the process of preparing brown rice.

The cosmetic composition of the present invention is characterized by containing a red rice extract.

The red rice extract may be prepared by extracting all or part of the red rice, but preferably may be a red rice rice bran extract prepared by extracting the rice bran portion of the red rice.

The red rice extract may be extracted according to a conventional plant extract manufacturing method, but is not limited thereto, and preferably in the group consisting of distilled water, 1,3-butylene glycol, alcohol having 1 to 4 carbon atoms, ethyl acetate, and acetone. It can be extracted with one or more solvents selected, preferably with metaols or ethanol, most preferably with ethanol. After extraction with the solvent, the filtration process may be further carried out, it may be concentrated under reduced pressure and / or lyophilized to powder. The solvent at the time of extraction may be extracted by adding 10 to 10,000 parts by weight based on 100 parts by weight of the extract, although not limited thereto.

The red rice extract is excellent in whitening activity by inhibiting melanin production by inhibiting the enzymatic activity of tyrosinase more effectively than white or brown rice (see Example 6). Arbutin is known to have particularly whitening activity. Compared with the red rice extract, while showing similar whitening activity, the cytotoxicity is very low and can be easily used as a cosmetic composition more safely than arbutin (see <Example 5>).

The cosmetic composition of the present invention may contain 0.001 to 20% by weight of the red rice extract, the content may be appropriately adjusted according to the content of the other ingredients contained in the formulation or cosmetic composition. However, when the content of the red rice extract in the cosmetic composition is less than 0.001% by weight, it may be difficult to expect a substantial whitening effect in the cosmetic composition, and when it exceeds 20% by weight, it may be uneconomical compared to the efficacy of the cost increase. Preferably, the red rice extract is included in an amount of 0.01 to 5.0 wt%. In addition, the cosmetic composition, in addition to the three kinds of extracts may include all kinds of ingredients usable in conventional cosmetics, such as excipients, diluents and the like.

The cosmetic composition of the present invention may be provided for the use of basic cosmetics, makeup cosmetics, body cosmetics, hair cosmetics, scalp cosmetics, shaving cosmetics or oral cosmetics. Examples of the basic cosmetics include creams, lotions, packs, massage creams, emulsions, etc. The makeup cosmetics include foundation, makeup base, lipstick, eyeshadow, eyeliner, mascara, eyebrow pencil, and the like. Examples include soaps, liquid cleansers, baths, sunscreen creams and sun oils. Hair care products include shampoos, rinses, hair treatments, hair mousses, hair liquids, pomades, hair colorants, hair bleaches, and colorants. There are rinses, hair tonics and scalp treatments as scalp cosmetics, aftershave and shaving creams as shaving cosmetics, and toothpaste and mouth washes as oral cosmetics. The cosmetic composition of the present invention is not limited thereto but is preferably skin external ointment, essence, lotion, cream, gel, lotion, pack, massage cream, latex, foundation, makeup base, soap, liquid cleanser, bath agent, sun cream, and It may be a formulation selected from the group consisting of sun oil. At this time, to improve the physical properties of the fragrance, pigments, fungicides, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, synthetic polymer materials and the like can be further added.

For example, when applied as an external skin ointment, in addition to the red rice extract may be prepared to contain 50.0 to 97.0% by weight of petrolatum and 0.1 to 5.0% by weight of polyoxyethylene oleyl-ether phosphate. In addition, when applied as a flexible softener, it may be prepared to contain 1.0 to 10.0% by weight of polyhydric alcohols such as propylene glycol, glycerin, and 0.05 to 2.0% by weight of surfactants such as polyethylene oleyl ether and polyoxyethylene cured castor oil. . In addition, when applied as a nutrient cosmetic cream or nourishing cream, in addition to the red rice extract 5.0 to 20.0% by weight of oils such as squalene, petrolatum, octyldodecanol and 3.0 to 15.0% by weight of wax components such as ethanol, stearyl alcohol, beeswax It can be prepared to contain. In addition, when applied as an essence, it can be prepared to contain 5.0 to 30.0% by weight of polyhydric alcohols such as glycerin, propylene glycol. In addition, when applied as a massage cream, in addition to the red rice extract may be prepared to contain 30.0 to 70.0% by weight of oil, such as liquid paraffin, petrolatum, isononyl isononanoate, and, if applied as a pack, A peel-off pack containing 5.0 to 20.0% by weight of vinyl alcohol or a wash-off pack containing 5.0 to 30.0% by weight of pigments such as kaolin, talc, zinc oxide and titanium dioxide in a general emulsified cosmetic. In addition, the composition for improving skin of the present invention is applied to the general cosmetic ingredients, such as oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc. as necessary. It is possible to.

The cosmetic composition of the present invention contains red rice extract, which effectively inhibits the enzymatic activity of tyrosinase and blocks melanin production by effectively inhibiting the enzymatic activity of tyrosinase and arbutin (arbutin), which is known to have excellent whitening activity. In comparison, the red rice extract exhibits similar whitening activity, but has very low cytotoxicity and can be easily used as a cosmetic composition more safely than arbutin.

1 is a picture of the shape of brown rice.
2 is a state of red rice used in the present invention.
3 is a rice bran extract of red rice prepared using 70% ethanol as a solvent. (A) Red rice (YW7758), (B) Brown rice YUM034), (C) White rice (YC0110).
4 is a chromatograph of free amino acid analysis results of rice bran extract of red rice (RdRc, YW7758).
FIG. 5 is a chromatographic view of a free amino acid analysis result of rice bran extract of brown rice (Rc, YUM034). FIG.
6 is a chromatographic diagram of the results of free amino acid analysis of rice bran extract of white rice (rdrc, YC0010).
Figure 7 is a graph comparing the content of gamma butyric acid (GABA) contained in the rice bran extract of red rice (RdRc YW7758), brown rice (YUM034), white rice (YUM044) by free amino acid analysis.
8 is a standard curve graph of GABA (γ-Amino Butyric acid).
Figure 9 is a graph of the content of GABA (γ-Amino butyric acid) of red rice rice, brown rice rice and white rice rice.
10 is a graph showing the cell survival test results of human skin keratinocyte line (HaCAT) treated with rice bran extract for 48 hours.
11 is a graph showing the cell survival test results of the human skin keratinocyte line (HaCAT) treated with rice bran extract for 72 hours.
Figure 12 shows the growth of cells by time of HaCAT cells taken at intervals of 24 hours, 48 hours, 72 hours, and 96 hours after treatment with 100 μg / ml of rice bran extract of red rice.
(A, C, E, G and I: untreated cells 0, 24, 48, 72, 96 hours growth, respectively, B, D, F, H and J: extract treated cells 0, 24, respectively Cell growth by 48, 72, 96 hours)
13 is a graph showing the effect of the extract of red and white rice on cell proliferation.
14 is a B16F10 melanin cell line of red rice bran extract It is a graph showing melanin inhibitory effect.
15 is red rice of rice bran extract It is a graph showing the inhibition rate of the enzyme activity of mushroom tyrosinase.
Figure 16 relates to the results of irradiation for 5 weeks the appearance stability test of moisturizing cream containing red rice bran extract.
Figure 17 relates to the results of a five-week irradiation of the stability test of the whitening cream containing red rice bran extract.
Figure 18 relates to the results of irradiation for 5 weeks the appearance stability test of the red rice bran extract extract containing sunscreen cream.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

< Reference Example  1>

Selection of Rice Used in the Invention

Rice used in the present invention is Rd or Rc RdRc as a weed or conventional rice collected from nature carrying genes or Rc genotype homozygous or heterozygous strains were used directly or rdrc recessive cultivars were used. Rice having a pigment-related gene composition and having a skin appearance of red, pale red, brown, and reddish brown was used as a sample to distinguish white or no pigment through genetic selection.

Genotypes and Varieties of Red, Brown and White Rice Color of the skin or seed genotype Breed name National species resource number Research Serial Number Red rice RcRd Red pearl 1-1-2001-5 YW7758 Brown rice Rcrd Gold tower 01-0001-2008-3 YW7760 White rice rcrd , rcRd course 01-0001-1997-21 YC0110

Specifically, as shown in Table 1, the red rice used in the present invention was harvested from rice of red pearl variety, and the genotype of rind was RcRd , and the brown rice used as comparative example was harvested from rice of gold tower variety. The genotype is Rcrd and the white rice is harvested from a variety of rice varieties and the genotype of the rind is rcrd or rcRd .

Rice bran used in this experiment was made into a fine powder using liquid nitrogen and stored at -80 ° C and used as a sample. 2 shows their color and appearance. Figure 2 shows the general name and genotype and phenotype of the pigment gene containing rice on the left side, in the present invention was added with reference to black rice.

< Example  1>

Red rice Rice bran  Preparation of Extract

To prepare rice bran extract, 70% ethanol corresponding to 20 times the amount of the rice bran sample prepared in <Reference Example 1> was added, followed by shaking at 30 ° C., RPM 170, in a culture incubator (KMC-8480 SF, Vision, Korea). Extraction was repeated twice under 12 hour conditions. Thereafter, three filtration processes were performed. After centrifugation with g-force 8,000 for 20 minutes, the filtrate was first filtered through a paper filter, followed by Advantec No. Secondary filtration to 2 and then filtered filtrate with a 0.2um advantec menbrane filter. The filtrate was completely removed from ethanol using a rotary pressure reducer (N-1000, Eyela, Japan) at 50 ° C., dried in a freeze dryer (FD-1, Eyela, Japan), crushed and stored at -80 ° C.

The extraction yield of 70% ethanol extract from 300 g of each type of rice bran described in <Reference Example 1> is shown in Table 2.

Specifically, 39g of red rice bran was obtained, and 31g of brown rice and 36g of white rice were extracted. Therefore, the extraction yield of the material from the dry rice bran weight was 13% for red rice, 10% for brown rice, and about 12% for white rice (Table 2). Figure 3 is a photograph of the extraction solution of each rice bran extracted with 70% ethanol. The extracts were concentrated, dried in lyophilizer and powdered.

Yield of rice bran extract of red rice, brown rice, white rice shape Rice bran / Brown rice (kg / kg) Rice yield
(%) Rice bran extract
/ Rice bran (g / g) Rice Yield Extract Yield (%) Red rice 3.57 / 30.42 11.72 39/300 13 Brown rice 1.18 / 8.2 14.39 31/300 10 White rice 3.16 / 34.56 9.15 36/300 12

< Example  2>

Red rice Rice bran  Determination of the content of phenolic substances in extracts

Phenolic compounds in plants are secondary metabolites and exhibit various physiological activities such as antioxidant and antimicrobial activity. In particular, antioxidant activity has been reported to work with phenolic compounds. Investigation of the content of phenolic compounds in plants is expected to be the primary data to explore the antioxidant activity of plant-derived natural extracts.

Specifically, the extraction method of total phenolic substances from rice bran was performed as follows. First, in order to obtain an extract, 10 ml of 70% ethanol is added to 0.5 g of rice bran and shaken at 10 ° C. for 1 hour. The supernatant obtained after centrifuging this at 8,000 rpm for 15 minutes at 4 degreeC was obtained by Whatman NO. It was used by filtration with 5 filter paper. 2.0 ml of 2% Na ₂ CO ₃ was added to 0.2 ml of each extracted sample solution, and after 2 minutes, 0.2 ml of 50% Folin-Ciocalteu's reagent was added thereto, and the mixture was left at room temperature for 30 minutes and absorbance was measured at 750 nm (Rhee et al. ., 1981, J. Food Sci . 46: 75-77). Content was calculated from the standard curve prepared using Gallic acid (0.5 mg / ml) (Kim et al., 2000, J. Korean). Soc . Food Sci . Nutr 29: 1127-1132).

In the present invention, the results of measuring the phenolic compound content of each rice bran extract are as follows. The polyphenol content per 1 g of fractions was 4.9 mg / g for red rice, 1.1 mg / g for brown rice and 1.3 mg / g for white rice (Table 3). In particular, red rice (YW7758) was found to be four times higher in total phenolic content than brown rice (YW7760) and white rice (YC0110).

Total phenolic content (mg GAE ^{1 )} / g-extract) contained in rice bran of red, brown and white rice Rice bran sample Total phenolic content Red rice ( RdRc , YW7758 (red pearl)) 4.9 Brown rice ( rdRc , YW7760 (gold tower)) 1.1 White rice ( rdrc , YC0110 (one article) 1.3

¹⁾ Polyphenol content was expressed as GAE (gallic acid equivalent) per gram of fraction

< Example  3>

Red rice Rice bran  Content Analysis of Free Amino Acids in Extracts

In the present invention, ethanol extract is prepared by extracting ethanol from red rice ( RdRc gene), brown rice ( Rc gene) and white rice and analyzing the free amino acid content using an amino acid analyzer, and the result is shown in FIG. 4 (red rice). , FIG. 5 (brown rice) and FIG. 6 (white rice) are shown and compared in Table 4.

Specifically, as shown in Table 4, the total free amino acid content was 2794.3 µg / g for white rice, 1514.7 µg / g for brown rice, and 1393.3 µg / g for red rice ( RdRc gene). The major constituent amino acids are aspartic acid, glutamic acid, alanine, and proline, and red rice was 288.3 ㎍ / g, 365.1 ㎍ / g, 168.5 ㎍ / g, 65.9 ㎍ / g, respectively. Μg / g and 70.3 μg / g white rice showed 403.5 μg / g, 1488.1 μg / g, 140.0 μg / g and 83.7 μg / g, respectively. Among the free amino acids, 7 kinds of essential amino acids, including threonine, valine, methionine, isoleucine, leucine, phenylalanine, and lysine, were included.The total essential amino acid content of each sample was 70.5㎍ / g red rice, 65.8㎍ / g brown rice, white rice 111.8 μg / g. Among them, the content of GABA was the highest in the red rice bran with RdRc gene ( 130 ug / g). Essential amino acids accounted for 5.1% of red rice, 4.3% of brown rice, and 4.0% of white rice, and the red rice bran with RdRc gene had the highest concentration of amino acids. Therefore, the utilization of red rice ( RdRc gene YW7758), a weed rice rice bran, as a functional material is considered to be good.

Free amino acid content in red, brown and white rice (㎍ / g-raw bran) Free amino acids Red rice (YW7758) Brown rice (YW7760) White rice (YC0110) Phosphoserine 7.7 ± 1.50 5.2 ± 0.77 8.3 ± 1.40 Taurine 19.2 ± 2.18 10.6 ± 2.75 30.3 ± 4.54 Phospho ethanol amine 23.2 ± 9.97 ^-1) - Urea - 34.7 ± 49.06 62.2 ± 88.01 Aspartic acid 288.3 ± 43.95 230.0 ± 23.52 403.5 ± 26.68 Throneine 25.9 ± 3.55 25.4 ± 1.38 38.4 ± 3.47 Serine 60.7 ± 6.21 39.8 ± 2.82 128.4 ± 6.30 Glutamic acid 365.1 ± 49.84 587.5 ± 57.82 1488.1 ± 121.03 Sarcosine - 8.3 ± 11.75 - α-amino adipic acid 0.8 ± 1.1 4.6 ± 0.75 4.3 ± 0.31 Glycine 29.5 ± 1.83 49.8 ± 6.17 20.6 ± 0.52 Alanine 168.5 ± 14.66 192.5 ± 8.64 140.0 ± 4.30 Citrulline 0.6 ± 0.84 0.7 ± 0.96 1.3 ± 1.85 α-amino-n-butyric acid 3.2 ± 0.84 1.3 ± 0.24 1.2 ± 0.12 Valine 26.0 ± 17.83 20.6 ± 4.01 30.8 ± 14.00 Methionine 1.1 ± 1.57 7.0 ± 2.56 10.2 ± 4.29 Cystathionine 3.0 ± 2.16 5.7 ± 2.54 7.2 ± 0.40 Isoleucine 4.9 ± 4.00 3.1 ± 4.36 8.1 ± 5.80 Leucine 3.8 ± 5.35 2.5 ± 3.51 8.5 ± 8.93 Tyrosine 8.4 ± 11.95 7.0 ± 9.85 12.1 ± 17.15 Phenylalanine - - 1.3 ± 1.83 β-Alanine 4.6 ± 0.35 4.1 ± 0.03 6.6 ± 0.69 β-Amino isobutyric acid 2.8 ± 0.22 4.4 ± 0.11 4.5 ± 1.02 γ-Amino-n-butyric acid 130.8 ± 15.68 30.3 ± 8.91 93.1 ± 12.48 Ethanol amine 24.3 ± 3.17 30.4 ± 10.90 37.2 ± 11.28 Ammonia 29.8 ± 2.62 11.3 ± 3.72 11.3 ± 4.54 Hydroxylysine - - - Ornithine 0.6 ± 0.83 1.8 ± 0.32 2.8 ± 0.96 Lysine 8.9 ± 2.60 7.3 ± 2.58 14.5 ± 4.33 1-Methylhistidine - 0.9 ± 1.25 2.3 ± 0.36 Histidine 18.0 ± 3.04 29.5 ± 4.53 27.1 ± 4.29 Anserine 0.0 3.9 ± 5.47 5.7 ± 8.00 Carnosine 20.8 ± 24.24 20.7 ± 29.22 24.1 ± 34.11 Arginine 46.9 ± 16.95 63.5 ± 19.45 76.5 ± 32.26 Proline 65.9 ± 13.2 70.3 ± 6.39 83.7 ± 16.68 TAA ^{2 )} 1393.3 1514.7 2794.3 EAA ^{3 )} 70.5 65.8 111.8 EAA / TAA (%) ⁴⁾ 5.1 4.3 4.0

¹⁾ Not detected

²⁾ TAA, total free amino acid

³⁾ EAA, essential aminoacid (Thr + Val + Met + Ile + Leu + Phe + His + Lys)

< Example  4>

Red rice Rice bran  Extract Of gamma aminobutyric acid  Quantitative Analysis

Gamma-amino acid-butyl; into free amino acids, which are widely distributed in nature (GABA γ- Amino-n-butyric acid) is known as GABA nerve excitement inhibitory neurotransmitter in the human brain (Inhibitory Neurotransmitter). In particular, GABA accounts for about 30% of the total neurotransmitter of the central nervous system and is present at a high concentration of about 200 to 1,000 times that of other neurotransmitters. In addition, GABA is involved in the regulation of many physiological mechanisms of the human body such as blood pressure increase suppression, vision enhancement, anti-anxiety, and anticonvulsion. GABA is also known to be effective in controlling the secretion of growth hormone and is effective in pain relief. It is a receiving substance. Therefore, in detail, in the present invention, the content of GABA contained in red and brown rice and brown rice of white rice was measured in detail.

<4-1> Free amino acids  According to the analysis result GABA Content of

The content of GABA was 130.8 ㎍ / g of red rice bran extract, 30.3 ㎍ / g of brown rice bran extract and 93.1 ㎍ / g of rice bran extract, of which 130.8 per 1 gram of rice bran ㎍ was much higher than other samples. This was found to be 4.3 times higher than brown rice of rice bran extract and 1.3 times higher than white rice (see FIG. 7).

<4-2> by enzyme method GABA Quantitative Analysis of

For the present invention, the quantitative analysis of GABA using enzymatic method was performed according to the method of Zhang and Bown (1997).

First, 0.1 g of finely ground rice bran and 400 ul of methanol were added to liquid nitrogen for pretreatment of the sample, and the mixture was left at 65 ° C. for 30 minutes. Thereafter, 1 ml of 70 mM LaCl ₃ was added to the sample, mixed, and centrifuged at 10,000 rpm for 5 minutes. 700ul of the supernatant of centrifugation and 160uL of 0.1M KOH were taken and stirred for 5 minutes. This was again used for the experiment supernatant centrifuged for 5 minutes at 10,000rpm.

Quantitative analysis of GABA was based on the Gabase enzyme quantification method of Sigma. A) 100 mM posassium pyrophosphate buffer (pH 8.6), B) 100 mM 2-Mercaptoethanol solution (2-ME), C) 25 mM β-Nicotinamide adenine dinucleotide phosphate solution (β-NADP), D) 10 mM γ-Amino- n-Butyric acid solution (GABA), E) 100 mM α-ketoglutarate solution, F) 75 mM potassium phosphate buffer with 25% (v / v) glycerol, pH7.2 (Enz-Dil), G) 1 units / ml Gabase Enzyme solution was prepared. After reacting the reagents A, B, E, C, and D for about 20 minutes at room temperature, G and F were added to measure absorbance 5 times at 60 seconds for 5 minutes at 340 nm using the total scan time of the spectrophotometer. By using the following formula to calculate the GABA content in rice bran through Equation 1 below and the experimental protocol is described in Table 5.

Quantitative Experiment Protocol of GABA Using GABAase Reagent Test (ml) Blank (ml) Control (ml) A (Buffer) 2.3 2.3 2.3 B (2-ME) 0.1 0.1 0.1 E (α-KG) 0.15 0.15 0.15 C (β-NADP) 0.15 0.15 0.15 D (GABA or sample) 0.3 (GABA) 0.3 (sample) 0.3 (Reagent A) Mix and leave for 10 ~ 20min G (GABase) 0.02 - - F (Enz Dil) - 0.02 0.02

8 and 9, the results of measuring the content of GABA components in the rice bran by the enzymatic method are as follows. The red rice (YW7758) contained 4.41uM of GABA, 2.0uM of brown rice (YW7760) and 2.4uM of white rice (YC0110) per 1g of rice bran sample.

The results showed that red rice had 1.8 times higher GABA content than white rice and 2.2 times higher GABA content than brown rice.

In the experiment of the present invention, the content of GABA in brown skin was less than that of white rice by analysis of free amino acid analysis and enzyme method. Therefore, the analysis result of the free amino acid (FIG. 7) and the analysis result of the enzyme method (FIG. 9) were in agreement.

< Example  5>

Red rice Rice bran  Cytotoxicity Test of Extract

Mouse-derived melanoma cells (B16F10, cell line number: KCLB80008) and human-derived keratinocyte cells (HaCAT) as cells used in the cytotoxicity test and growth in vivo melanin inhibition test used in the present invention were received from Korea Cell Line Bank (KCLB). Incubated.

The culture was incubated with DMEM (Dulbecco's modified Eagle's medium, sigma) containing 10% FBS and 2% penicillin / streptomycin in a cell culture medium maintained at 37 ° C., 5% CO 2 and constant humidity. The culture was exchanged at intervals of 2 to 3 days and subcultured.

The effects of rice bran extract, the skin of red rice, on the cytotoxicity and proliferation of HaCAT cells, which are keratynocytes derived from human skin, were investigated.

Specifically, after culturing for 48 and 72 hours by adding rice bran extract of various concentrations to the cell culture solution, MTT assay was performed and cell survival results were examined.

In MTT assay, cultured HaCAT cells were dispensed in 96 well plates at 1 × 10 ⁴ / well, and after 24 hours, each rice bran extract was dissolved in 0.1% DMSO and finally treated with cells at 25, 50, and 100 ㎍ / ml. It was. After 48 hours and 72 hours of sample treatment, the supernatant was removed and 90 μl of DMEM media was added, followed by MTT (3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H prepared at 2 mg / ml. 10 μl of a solution of -tetrazolim bromide, Sigma, USA) was added to each well, blocked with foil, and reacted for 4 hours in a 37 ° C., 5% CO ₂ incubate. After removing the supernatant and treating 200 μl of DMSO, absorbance was measured at 540 nm using an ELISA microplate reader (Emax, Molecular devices, USA).

<5-1> Red rice Rice bran  If the extract was treated for 48 hours

Experimental results when the red rice bran extract was treated for 48 hours are shown in FIG. 10. In experiments treated with rice bran extracts of red rice (YW7758), white rice (YC0110), brown rice (YW7760) and GABA for 48 hours, a group containing only 0.1% DMSO was used as a control.

As shown in FIG. 10, the red rice bran extract (YW7758) 25ug / ml treated group had a cell survival rate of 107.8% and a cell survival rate of 102.8% at a concentration of 50ug / ml and a concentration of 100ug / ml compared to the control group. The survival rate was 98.1%. Therefore, they did not show toxicity to cells, but rather induced cell proliferation.

The brown rice bran extract (YW7760) showed 108.6% cell growth at 25ug / ml and 104.3% at 50ug / ml, and 95.4% at 100ug / ml. The rice bran extract group (YC0110) of white rice showed 103.4% cell viability at 25ug / ml concentration, 100.7% cell viability at 50ug / ml concentration, and 104.8% cell viability at 100ug / ml concentration. It was confirmed that more than 90% induces safety and cell proliferation for normal cells. In general, the cell survival rate was 107.5% at 25ug / ml concentration, 105.5% at 50ug / ml concentration, and 99.4% at 100ug / ml concentration.

As a result, the growth rate of the cells according to the concentration was similar to that of the rice bran samples of red rice (YW7758), brown rice (YW7760), and white rice (YC0110) and GABA. Able to know.

<5-2> Red rice Rice bran  When the extract was treated for 72 hours

Experimental results when the red rice bran extract was treated for 72 hours are shown in FIG. 11.

As shown in FIG. 11, the red rice bran extract group had a cell viability of 107.8% at a concentration of 25 ug / ml, a cell survival rate of 105.9% at a concentration of 50 ug / ml, and a cell survival rate of 98.4 at a concentration of 100 ug / ml compared to the control group. % Did not show toxicity to the cells, but rather induced cell proliferation.

In addition, the brown rice bran extract group had a cell survival rate of 101.7% at 25ug / ml concentration, a cell survival rate of 107.9% at a concentration of 50ug / ml, and a 93% cell survival rate at 100ug / ml.

In the case of white rice, the cell survival rate was 105.4% at 25ug / ml concentration, 103.1% cell survival rate at 50ug / ml concentration, and 100.6% cell survival rate at 100ug / ml concentration. It was confirmed that the induction of cell safety and cell proliferation.

In the case of GABA treatment, the cell survival rate was 101.1% at 25ug / ml concentration, 99.9% cell survival rate at 50ug / ml concentration, and 97.5% cell survival rate at 100ug / ml concentration. Red and brown and white and GABA showed similar results

<5-3> Red rice Rice bran  Cell growth observed when treated with extract

In the present invention, when the extract is used as a cosmetic, the effect on the growth and growth of human skin cells is important, the effect on the growth of keratinocytes was investigated.

MTT assay was performed for 24 hours, 48 hours, 72 hours, after treatment with red rice bran extract and GABA at different concentrations (0 μg / ml, 25 μg / ml, 50 μg / ml, 100 μg / ml). Hourly cell growth of HaCAT cells was recorded at 96 hour intervals, and the results are shown in FIG. 12.

As shown in FIG. 12, cell viability and growth were normal in most cases.

<5-4> Red rice Rice bran  Investigation of the Effects of Extracts on Cell Growth and Proliferation

The effects of RdRc gene-expressing red rice (YW7758) and rice bran extracts on HaCAT cells, which are human skin keratinocytes, were investigated for their growth and proliferation. The final concentration of the red rice extract was treated to 100 μg / ml in HaCAT cells, and after 0 hours, 48 hours, 96 hours, 144 hours, 216 hours, and 264 hours, trypan blue exclusion test was performed. Numerical increase was counted. Brown rice was excluded from the growth rate because it showed similar amino acid composition and survival rate.

After the sample treatment, tryptophan blue was administered and the results of counting the living cells are shown in FIG. 13.

As shown in FIG. 13, the cell number increased from 0 hours to 96 hours after treatment with brown rice extract, and cell proliferation rapidly increased by 144 hours, and cell proliferation was increased by 216 hours. . After 216 hours after the extract treatment, the total amount of cells in the red rice (YW7758 or RdRc gene) -treated extract was about 30% higher than the untreated control (0.1% DMSO). The total cell number tended to increase by 25%. After 216 hours after the sample treatment, the growth rate of the control and all cells decreased and the survival rate of HaCAT cells treated with the extract of red rice was the highest.

In conclusion, the extracts of red rice (YW7758) and white rice (YC0110) promoted cell growth. Table 6 is a table showing the average of the actual cell count and the standard deviation three times the experiment.

Effects of Rice Bran Extracts from Red and White Rice on the Proliferation of HaCAT Cells No treatment No treatment Red rice White rice 0 0.1% DMSO 0.1% DMSO 0.1% DMSO 0hr 10000 10000 10000 10000 48hr 28750 22500 28750 26250 96hr 63750 58750 80000 75000 144hr 468750 471875 690625 628125 216hr 1687500 1792500 2257500 2058750 264hr 1267500 1312500 1503750 1402500

< Example  6>

Red rice Rice bran  Inhibition and Whitening Activity of Extracts by Melanin Production

In the present invention, the effect of inhibiting the melanin production of the extract from B16F10, the skin melanosite of the mouse in order to determine the rate of inhibition of melanin formation caused by pigmentation of red rice bran in the cell.

<6-1> Melanin production inhibitory effect

Incubate the cultured B16F10 cells into 6-well plates at 5x10 ⁴ per well and stabilize for 24 hours at 37 ° C and 5% CO ₂ conditions, then white rice ( rdrc , YC0110), red rice ( RdRc , YW7758), Arbutin ( sigma, USA) was dissolved in 0.1% DMSO and diluted with DMEM and treated to a final concentration of 0.01, 0.1, 0.25, 0.5%, 37 ℃, 5% CO ₂ The incubator was incubated for 3 days.

The supernatant of the culture solution was removed and the cells attached with trypsin EDTA were separated and placed in a 1 ml DMEM medium to distribute the cells evenly with a pipette. 20 µl of the suspension was mixed with 20 µl of 0.4% trypan blue (Gibco, USA) solution, and the cells were stained. After counting, 50,000 cells were taken, centrifuged at 1,500 rpm for 3 minutes, transferred to a micro tube, and centrifuged at 12,000 xg for 3 minutes. After drying for 1 minute in the heat bloc of 1M NaOH and 10% DMSO solution was put into a 500ul pipette. Melanin was dissolved in an 80 ° C. water bath for 20 minutes, centrifuged at 12,000 rpm for 3 minutes, and absorbance was measured at 490 nm, and the melanin production inhibition rate was calculated using Equation 2 below.

The B16F10 melanocytes were treated with rice bran extract at a final concentration of 50 µg / ml and 100 µg / ml for 3 days, and then cell viability and melanin production were measured. Each experiment was repeated three times.

As shown in FIG. 14, the extract of red rice (YW7758) inhibited melanin production by 29% at 50 μg / ml treatment, and melanin production was suppressed by 45% at 100 μg / ml. On the other hand, melanocytes grown in 0.1% DMSO without the control extract showed no inhibition of melanogenesis. In addition, as shown in Figure 13, the treatment of the extract of red rice bran did not affect the growth of cells. However, it can be seen that melanin production is inhibited in a concentration dependent manner. However, in the case of white rice (YC0110), melanin production was suppressed by about 16% at the extract concentration of 50µg / ml, and melanin production was inhibited by 33% at 100µg / ml. However, melanin production of red rice bran extract was higher than that of white rice bran extract.

In addition, as compared with arbutin, a representative whitening material, it can be seen that the rice bran extract of red rice is similar to arbutin in its ability to inhibit melanin production. In particular, when using a high concentration of arbutin due to the toxicity of the cell can not be used in high concentrations in the composition of the cosmetics, red rice extract is considered safe because it is safe to use.

<6-2> Tyrosinase  Enzyme activity inhibition

Tyrosinase or tyrosinase is an enzyme involved in the melanin biosynthesis pathway in the human body and the whitening components have a mechanism of action to inhibit this enzyme. Tyrosinase converts tyrosine to L-DOPA which is then converted to dopaquinone and finally produces melanin.

Therefore, in order to evaluate the degree of inhibition of tyrosinase enzyme activity in vitro, red and white ethanol extracts were dissolved in a solvent and diluted to 0.001, 0.01, 0.1, 0.25, 0.5%, and then 0.1M phosphate buffer solution (pH6.8). 250ul, 225ul of sample solution and 25ul of mushroom Tyrosinase (2,000U / mL) solution were added in this order. Put 1.5mM Tyrosin 250ul into the solution and reacted for 20 minutes at 37 ℃ and left for 5 minutes on ice to stop the reaction. Thereafter, the absorbance at 490 nm was measured using a spectrophotometer, and the degree of enzymatic activity inhibition of tyrosinase was measured through Equation 3 below. As a control, 0.1M phosphate buffer (pH 6.8) was used instead of the sample solution. As a positive control, arbutin (Arbutin, hydroquinone-beta-d-glucopyranoside) was diluted to the same concentration as red rice bran (YW7758) and white rice bran (YC0110).

 The results of the whitening experiments are shown in Table 7 and FIG. 15. As shown in Table 7 and FIG. 15, as a result of analyzing the tyrosinase inhibition effect, the inhibitory effect of tyrosinase activity was hardly observed in 0.001% of arbutin as a positive control, but in 0.01, 0.1, 0.25, and 0.5%, respectively. 42, 81, 95, 98% showed a concentration-dependent inhibitory effect of tyrosinase activity.

In addition, red rice bran extract (YW7758) increased the tyrosinase activity inhibitory effect in a concentration-dependent manner, inhibited 50% at 0.25% concentration, but inhibited tyrosinase activity by 75.6% at 0.5% concentration.

On the other hand, white rice (YC0110) rice bran extract also showed a concentration-dependent inhibitory effect on tyrosinase activity, but showed an inhibitory effect of about 50% at the highest concentration of 0.5%, but significantly lower than red rice bran extract (YW7758). Effect.

Inhibitory Effect of Rice Bran Extract on Tyrosinase Activity (Average Error Value of Three Repeats) YC0110
(White rice) 0.001% 0.01% 0.1% 0.25% 0.5% 1set 4.970 2.434 14.300 28.905 53.550 2set 5.469 2.100 13.514 29.574 44.994 3set 5.832 0.648 14.255 33.369 43.413 AVERAGE 5.423 1.727 14.023 30.616 47.319 STDEV 0.433 0.950 0.442 2.408 5.454 YW7760
(Brown taste) 0.001% 0.01% 0.1% 0.25% 0.5% 1set 0.059 2.309 6.758 21.090 38.709 2set 0.268 3.998 10.213 18.260 40.256 3set 0.382 3.250 7.457 19.329 44.157 AVERAGE 0.236 3.186 8.143 19.560 41.041 STDEV 0.164 0.846 1.827 1.429 2.807 YW7758
(Red taste) 0.001% 0.01% 0.1% 0.25% 0.5% 1set 6.187 18.458 30.325 56.897 73.935 2set 5.580 14.352 34.719 52.589 78.021 3set 5.508 6.479 28.942 46.652 75.810 AVERAGE 5.758 13.097 31.329 52.046 75.922 STDEV 0.373 6.087 3.017 5.144 2.045 Arbutin
(Arbutin) 0.001% 0.01% 0.1% 0.25% 0.5% 1set 0.710 20.183 58.519 95.740 98.986 2set 2.009 14.469 57.173 85.270 95.984 3set 2.808 19.438 55.940 81.210 94.276 AVERAGE 1.842 18.030 57.210 87.407 96.416 STDEV 1.059 3.106 1.290 7.497 2.384 Overall average 0.001% 0.01% 0.1% 0.25% 0.5% YC0110
(White rice) 5.423 1.727 14.023 30.616 47.319 YW7760
(Brown taste) 0.236 3.186 8.143 19.560 41.041 YW7758
(Red taste) 5.758 13.097 31.329 52.046 75.922 Arbutin
(Arbutin) 1.842 18.030 57.210 87.407 96.416

< Manufacturing example >

Cosmetics  Preparation of the composition

Using the red rice extract of the present invention to prepare a composition of the cosmetic. General composition Whitening cream, moisturizing cream, was prepared by adjusting the concentration of the red rice extract in the cream block.

< Manufacturing example  1> Preparation of Lotion

Lotions containing red rice bran extract Prize Cosmetics % wt. A Cetyl PEG / PPG Dimethicone 2.50 Polyglycerin-4-isostearate 1.20 Cetyldimethicone 1.00 Eten Homopolymer 0.50 Dicapyryl Chamonate 5.00 Cocoglycerides 3.00 Cetyloctaneate 5.00 B Purified water control Propylene glycol 5.00 D-panthenol 0.50 Sodium Chlorary 1.00 C Cyclopentasilaxen 8.00 Mica / titanium dioxide / tin oxide 2.10 D Tocopherol Acetate 0.50 Red rice  extract 2.00 Gamma Aminobutyl Acid 0.10 direction q.s. antiseptic q.s.

To prepare the lotion, as shown in Table 8 above, Phase A was heated to 80-85 ° C., Phase B was heated to 80 ° C. and stirred while slowly adding Phase B to A. Phase C was added and then mixed into Phase (A + B) until well dispersed. The stirred composition was cooled to 40-50 ° C., and then phase D was added and mixed with slow stirring. The composition was stirred while cooling to 35-40 ° C.

< Manufacturing example  2> Moisturizing cream  Produce

Moisturizing cream containing rice bran extract of red rice Chemical name content(%) Purified water to100 Trimethylolpropane tricaprylate / caprate 4 Propylene glycol 2 Betaine One Polysorbate 60 One Fiji / Fiji-17 / 6 copolymer One Stearic acid 0.7 Cyclomethicone 0.5 Glyceryl Stearate / Pig-100stearate 0.5 Dimethicone 0.5 Dimethicone / Vinyl Dimethicone Crosspolymer 0.3 Sorbitan stearate 0.30 Imidazolidinyl urea 0.2 Methylparaben 0.2 Allantoin 0.1 Collagen 0.1 Xanthan Gum 0.1 Sodium DieA 0.1 Butyl Paraben 0.1 NATO GUM 0.1 Tocopheryl acetate 0.05 Disodium ID 0.02 Sodium hyaluronate 0.01 Yellow No. 6 0.01 Combination spices 0.1 Red rice  extract 0.5-2.0

The moisturizing cream was prepared in the same manner as in <Preparation Example 1>.

< Manufacturing example  3> Sunscreen  Produce

Sun Cream with Red Rice Bran Extract Prize CTFA INCI Naming % I Solbitan Oliveate 3.00 Seteas-6 Olivet 1.50 Dibutyl adipate 3.00 Octocrylene 5.00 Diethylamino hydrobenzoyl hexabenzoate 2.00 Ethylene Methoxycinate 10.00 Netoshiphenyl Trazaine 2.00 II Purified water Up to 100.00 glycerin 3.00 Sodium Carbamer 0.30 Methene Bisbenzotriazole Tetramethylbutylphenol 6.00 Gamma aminobutyric acid 0.5 Red rice  extract ~ 2.00 III perfume a.n. antiseptic a.n.

< Experimental Example  1>

Of the present invention Cosmetics  Appearance safety of the composition

In order to study the safety of the developed cosmetics using the raw materials developed in the present invention was tested in accordance with the safety guidelines test standards of the Agency.

A cosmetic composition was prepared by adding rice bran extract and GABA of white rice (YC0110) and red rice (YW7758). The temperature safety test of the basic composition used the method of observing or measuring the appearance and physical properties of the product after leaving the contents of cosmetics according to the temperature conditions in the distribution process and the form of consumer use. : Color difference, fading, stripe, color stain, alien substance mixing, wound, float, separation, sedimentation, sweating, powder, crack, gelling, transparency, caking, bubble, pinhole, lifting, mold, oozing, lowering, container It was a measurement such as a change in incense in the inside and a change in use. The physical properties were measured by measuring pH, hardness, viscosity, turbidity, emulsification and powder particles diameter, loss of moisture drying, emulsification type, softening point and melting point.

<1-1> Moisturizing cream  stability

As shown in Figure 16, the stability of the moisturizing cream prepared with red rice bran extract was investigated. After 5 weeks of safety of the moisturizing cream containing red rice bran extract, no degeneration or unusual physical changes were observed.

<1-2> stability of whitening cream

As shown in Figure 17, the stability of the whitening cream prepared with red rice bran extract was investigated. Investigation of the safety of the whitening cream containing red rice bran extract over time for 5 weeks showed no degeneration or unusual physical changes.

<1-3> Sunscreen  stability

As shown in Figure 18, the stability of the sunscreen prepared with red rice bran extract was investigated. As a result of 5 weeks of safety of the sunscreen containing red rice bran extract, no degeneration or unusual physical changes were observed.

< Experimental Example  2>

Of the present invention Cosmetics  Of the composition pH  Safety investigation according to the change

After preparing whitening cream, sunscreen cream and moisturizing cream mixed with red and white rice extract samples, the pH stability of the composition was examined for 5 weeks. As shown in Tables 10, 11 and 12, there was no change in pH.

<2-1> Moisturizing cream  stability

PH change of moisturizing cream after adding red and white rice rice bran extract (25 ℃) Experimental product Day 1 2nd week 3rd week 4 weeks Week 5 result Control 6.74 6.72 6.75 6.74 6.73 Good Moisturizing Cream + YC0110 6.67 6.65 6.66 6.68 6.67 Good Moisturizing Cream + YW7758 6.7 6.68 6.71 6.7 6.69 Good Moisturizing Cream + YW7758 / GABA 6.69 6.69 6.67 6.71 6.69 Good

* White Rice (YC0110), Red Rice (YW7758)

<2-2> stability of whitening cream

PH change of whitening cream after adding red and white rice rice bran extract (25 ℃) Experimental product Day 1 2nd week 3rd week 4 weeks Week 5 result Control 7.38 7.37 7.38 7.36 7.39 Good Whitening Cream + YC0110 7.19 7.2 7.18 7.19 7.2 Good Whitening Cream + YW7758 7.12 7.11 7.13 7.12 7.11 Good Whitening Cream + YW7758 / GABA 7.17 7.16 7.18 7.17 7.16 Good

* White Rice (YC0110), Red Rice (YW7758)

<2-3> Sunscreen  stability

PH change of sunscreen after adding red and white rice bran extract (25 ℃) Experimental product Day 1 2nd week 3rd week 4 weeks Week 5 result Control 5.53 5.53 5.55 5.56 5.53 Good Sun Cream + YC0110 6.28 6.25 6.27 6.29 6.28 Good Sun Cream + YW7758 5.72 5.72 5.73 5.71 5.7 Good Sun Cream + YW7758 / GABA 5.55 5.55 5.56 5.54 5.55 Good

* White Rice (YC0110), Red Rice (YW7758)

< Experimental Example  3>

Of the present invention Cosmetics  Thermal safety investigation of the composition

The thermal stability at 40 ° C. of the composition mixed with the rice bran extract of red and white rice was investigated.

After preparing a whitening, sunscreen and moisturizing cream, the thermal stability of the composition was examined for 5 weeks. As shown in the table below, there was no change in physical properties.

Thermal Safety Study of Moisturizing Cream Containing Red and White Rice Bran Extract (CPI) Experimental product Day 1 2nd week 3rd week 4 weeks Week 5 result Control 400000 350000 230000 280000 250000 Good Moisturizing Cream + YC0110 185000 156000 150000 200000 280000 Good Moisturizing Cream + YW7758 450000 430000 190000 260000 300000 Good Moisturizing Cream + YW7758 / GABA 230000 215000 160000 250000 200000 Good

* White Rice (YC0110), Red Rice (YW7758)

Thermal Safety Study of Whitening Cream Containing Red and White Rice Bran Extract Experimental product Day 1 2nd week 3rd week 4 weeks Week 5 result Control 865000 840000 660000 720000 780000 Good Whitening Cream + YC0110 870000 830000 780000 850000 880000 Good Whitening Cream + YW7758 868000 620000 600000 750000 680000 Good Whitening Cream + YW7758 / GABA 900000 760000 730000 850000 885000 Good

Thermal Safety Investigation of UV Blocking Cream Containing Red and White Rice Bran Extract Experimental product Day 1 2nd week 3rd week 4 weeks Week 5 result Control 405000 300000 275000 255000 205000 Good Sun Cream + (YC0110) 350000 245000 250000 225000 220000 Good Sun Cream + (YW7758) 350000 300000 295000 235000 245000 Good Sun Cream + (YW7758) / GABA 375000 280000 285000 295000 225000 Good

* White Rice (YC0110), Red Rice (YW7758)

Claims (6)

Cosmetic composition for skin whitening containing red rice extract The cosmetic according to claim 1, wherein the red rice is rice of red pearl variety (National Species Variety Registration No. 1-1-2001-5) or red pearl wine variety (National Species Resource Variety Registration No. 1-1-2007-7) Composition. The cosmetic composition according to claim 1, wherein the red rice extract is extracted from rice bran of red rice. The cosmetic composition of claim 1, wherein the red rice extract is extracted with one or more solvents selected from the group consisting of distilled water, 1,3-butylene glycol, alcohols having 1 to 4 carbon atoms, ethyl acetate, and acetone. The cosmetic composition of claim 1, wherein the cosmetic composition is a skin external ointment, essence, lotion, cream, gel, lotion, pack, massage cream, latex, foundation, makeup base, soap, liquid cleanser, bath, sun cream, and sun oil. Cosmetic composition having a formulation selected from the group consisting of. The cosmetic composition of claim 1, wherein the cosmetic composition contains 0.001 to 20% by weight of red rice extract based on the total composition.

Images