KR101121980B1 - Manufacturing Method for Extracts of Leaves And the Leaves Using the Same and Cosmetic Composition Containing the Same - Google Patents

Abstract

The present invention relates to a lotus leaf extract manufacturing method, and more particularly to a lotus leaf extract manufacturing method comprising the step of enzymatic treatment by adding a cell wall degrading enzyme. Lotus leaf extract production method according to the present invention has a high polyphenol content, high whitening activity such as tyrosinase inhibitory effect and melanin production inhibitory effect, excellent anti-aging effect such as DPPH radical scavenging activity and SOD-like activity There is an effect that can provide a lotus leaf extract.

The present invention also has the effect of providing a cosmetic composition having a whitening effect and an anti-aging effect by containing the lotus leaf extract as an active ingredient.

Lotus leaf, extract, enzyme, cosmetic composition

Description

Manufacturing Method and Extracts of Leaves and the Leaves Using the Same and Cosmetic Composition Containing the Same}

The present invention relates to a lotus leaf, and more particularly, to a method for producing a lotus leaf extract having a high yield of extract of lotus leaf, and having a whitening activity and anti-aging effect, and a lotus leaf extract and a cosmetic composition containing the same.

In modern society, interest in skin care is increasing due to the improvement of living standards and the extension of the average life span, and interest in skin whitening is increasing due to the increase of the photoaging of the skin due to the increase of UV exposure of the skin due to environmental pollution. . Accordingly, researches on the development of whitening products using natural products are being actively conducted.

Whitening agents have the effect of inhibiting melanin synthesis by inhibiting melanocyte growth or inhibiting tyrosinase activity. Melanin is one of the pigments involved in the color of skin, hair and eyes (Li W, Hill HZ, Induced melanin reduces mutations and cell killing in mouse melanoma. Photochem Photobiol. 1997; 65 (3): 480-485).

Inhibiting melanogenesis controls copper (Cu), an essential component of the active site of tyrosinase, an enzyme involved in melanogenesis, or tyrosinase related protein I (TRP-1). And iron ion (Fe), an essential component of tyrosinase related protein II (TRP-2), to inactivate enzymatic activity or to inhibit the production of melanin intermediates. (NEWS & INFORMATION FOR CHEMICAL ENGINEERS, Vol. 25, No. 2, 2007)

Human skin secretion of hormones that regulate metabolism decreases internally over time, and when skin is exposed to ultraviolet rays, melanocytes develop immediate or delayed pigmentation depending on their wavelength. This includes freckles, blemishes, milk coffee spots, birthmarks, and surpluses.

In the skin exposed to sunlight, ultraviolet rays generate reactive oxygen species (ROS). As a result, the risk of photo-oxidative damage of ROS-derived skin is substantially increased, and when these excess free radicals are produced in vivo, mutations, inflammation, cell killing, carcinogenesis and skin aging appear. (Antioxidative properties of Baicalein Component from Scutellaria baicalensis George and Its Application to Cosmetics (Ⅰ), J. Korean Ind. Eng. Chem, Vol. 14, No. 5, August 2003, 657-665)

Yeon (蓮, Nelumbo nucifera Gaertiner) is a perennial aquatic plant of the water lily family, lotus leaf of the lower leaf, lotus flower softening, the operation of lotus is called Yeonsu.

Lotus leaf chemicals include roemerine, nuciferine, nornuciferine, armepavine, pronuciferine, and N-nornuciferine ), DN-methylcoclaurine, anonaine, liriodenine, quercetin, isoquercitrin, nelumboside, tatar Acids (tataric acid), malic acid (malic acid), gluconic acid (gluconic acid), acetic acid (acetic acid), succinic acid (succinic acid) and the like are known. In addition, lotus leaf is known to treat sequence alternating, hemostatic effect on hemostasis and hypothermia.

In recent years, the range of applications of lotus leaf, such as food and pharmaceutical fields using natural products in consideration of stability is wide. Therefore, in the present invention, a method of preparing a lotus leaf extract excellent in whitening and anti-aging effects such as increasing the extraction yield of lotus leaf, antioxidant effect, tyrosinase activity inhibitory effect, melanin production inhibitory effect.

The present invention provides a method for producing a lotus leaf extract and a lotus leaf extract according to the use of cell wall degrading enzymes in extracting lotus leaf, not only to increase the extraction efficiency of the active ingredient of lotus leaf, but also to have a high polyphenol content, better whitening activity and a higher antioxidant effect. It aims to do it.

The present invention also provides a cosmetic composition having a whitening effect and an anti-aging effect by containing the lotus leaf extract as an active ingredient.

The present invention for achieving the above object comprises the steps of preparing a (蓮, Nelumbo nucifera Gaertner ) leaf; Adding distilled water to the prepared lotus leaf and adjusting pH; After the pH adjustment, enzymatic treatment by adding cell wall degrading enzyme; And after the enzyme treatment, filtration or centrifugation provides a lotus leaf extract manufacturing method comprising the step of obtaining a lotus leaf extract.

In the present invention, the degrading enzyme is characterized in that the enzyme is a beta-glucanase, cellulase (celullase), proteinase (proteinase), alpha-amylase (α-amylase) and pentosanase (pentosanase) Provides a method of producing lotus leaf extract.

In addition, the present invention provides a method for producing lotus leaf extract, characterized in that the enzyme is pullulanase (pullulanase).

In addition, the present invention provides a method for producing lotus leaf extract, characterized in that the concentration of the enzyme is 1 ~ 2% (w / w).

The present invention also provides a lotus leaf extract lotus leaf extract prepared according to the above method having a polyphenol content of 1,100-1,500 µg / ml.

The present invention also provides a whitening and skin anti-aging cosmetic composition containing the lotus leaf extract as an active ingredient.

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

Lotus leaf extract preparation method of the present invention has a high polyphenol content, high whitening activity such as tyrosinase inhibitory effect and melanin production inhibitory effect, DPPH (1,1-diphenyl-2-picrydrazyl (1.1-diphenyl-) 2-picryl-hydrazyl)) has the effect of providing a lotus leaf extract excellent in anti-aging effects such as radical scavenging and SOD-like activity.

The present invention is also prepared according to the method of the present invention, by containing the lotus leaf extract excellent in whitening activity and anti-aging effect as an active ingredient has an effect that can provide a cosmetic composition having a whitening and anti-aging effect.

Hereinafter, with reference to the accompanying drawings will be described in detail the lotus leaf extract manufacturing method according to the present invention.

1 is a flow chart showing a lotus leaf extract manufacturing method according to an embodiment of the present invention.

First, the lotus leaf extract manufacturing method according to the invention is prepared by drying the lotus leaf (S110). After the lotus leaf is dried, it is preferable to grind to a suitable size, for example, the size of 50 to 200 mesh, and most preferably, about 100 mesh to increase the extraction yield of the lotus leaf active ingredient.

Then, distilled water is added to the prepared lotus leaf (S120). The reason for adding distilled water to the lotus leaf as described above is to make the enzyme easy to work. The amount of distilled water added is preferably about 20 to 30 times the volume of the prepared lotus leaf.

After adding the distilled water to adjust the pH (S130). The pH is preferably adjusted to the most active pH 5 ~ pH 8, the enzyme is not particularly limited when adjusting the pH, it is preferable to use 1M HCl and 0.1N NaOH.

After adjusting the pH of the distilled water containing the lotus leaf as described above is subjected to the enzyme treatment by adding the cell wall degrading enzyme (S140). The active ingredients of the leaves of the plant are known to be contained in the cells, and since the leaves are mainly composed of fibrin, they can be degraded by the cytolytic enzyme of the plant. In other words, the enzyme treatment is to act as a plant cell enzyme to break down the cell wall of the lotus leaf so that the effective ingredient is well extracted.

Preferred cell wall degrading enzymes for use in the lotus leaf extract, the production method of the present invention is pen tryptophan ^TM (PENTOPN), serenity mix ^TM (CEREMIX), Hatori atom ^TM (VISCOZYME), Ultra Flow ^TM (ULTRAFLO), pekti Annex ^TM (PECTINEX ), cellulose Cloud host ^TM (CELLUCLAST), Tunisia Carr claim ^TM (TUNICASE), promotional atom ^TM (PROMOZYME) and the like. In the above, pentophan ^TM (PENTOPN) is a single enzyme of xylanase, and ceremix ^TM (CEREMIX) is a beta-glucanase (β-glucanase) cellulase (cellulase), proteinase (proteinase) , Four enzymes, such as alpha-amylase and pentosanase, are mixed in the same weight ratio, respectively. Biscozyme ^TM (VISCOZYME) is a beta-glucanase (β-glucanase) Cellulase, hemicellulase, xylanase, arabanase and other enzymes are mixed in the same weight ratio, respectively.Ultraflow ^TM (ULTRAFLO) is an enzyme -Glucanase (β-glucanase), enzymes (Cellulase), xylanase (Xylanase), pentosanase (pentosanase), arabanase (arabanase), each of the four enzymes mixed in the same weight ratio to be. In addition, Pectinex ^TM (PECTINEX) is a polygalacturonase, pectine esterase, cellulase, hemicellulase, and the like in Pectintranseliminase. Enzyme of each species in the same weight ratio, Cellullast ^TM (CELLUCLAST) is the cellulase (Cellulase) single enzyme, Tunicase ^TM (TUNICASE) is a beta-glucanase (β-glucanase) single enzyme Promozyme ^TM is a Pullulanase homoenzyme.

The enzymes may be used alone or in combination of two or more, and is not particularly limited, but is preferably added in an amount of 1 to 2% (w / w).

After the addition of the degrading enzyme as described above, in order to further increase the extraction yield, it is preferably immersed for 1 to 4 hours at a temperature of 30 ~ 70 ℃.

Then, after the enzyme treatment as described above is filtered or centrifuged to obtain the lotus leaf extract of the present invention (S150). At this time, for example, Whatman No. It is preferable to filter by 41, and the centrifugation is not particularly limited, but centrifugation for 20 to 30 minutes is preferred at a centrifugal force of 2000 × G or more.

According to the method of producing a lotus leaf extract using the enzyme according to the present invention, the extraction yield of the lotus leaf extract is high, the polyphenol content is high as about 1,100 ~ 1,500㎍ / ㎖, to obtain a lotus leaf extract with more excellent whitening activity and antioxidant effect It can work.

Accordingly, the lotus leaf extract prepared according to the method of the present invention has a high content of polyphenol as an active ingredient of about 1,100 to 1,500 µg / ml, and is excellent in inhibiting tyrosinase activity and inhibiting melanin production, and has an excellent whitening effect. There is. In addition, the lotus leaf extract has a high DPPH radical scavenging activity and SOD-like activity, has excellent antioxidant activity, that is, anti-aging effect.

In addition, the lotus leaf extract was confirmed that there is no problem in stability when applied to the skin.

Next, the present invention provides a cosmetic composition containing the lotus leaf extract prepared according to an embodiment of the present invention as an active ingredient. Whatever the cosmetic composition is prepared by any method, if it contains a lotus leaf extract prepared according to an embodiment of the present invention corresponds to the present invention.

The cosmetic composition containing the lotus leaf extract prepared according to the embodiment of the present invention as an active ingredient is very stable because there is almost no change in pH according to temperature change. Even under natural light exposure, there is almost no change in pH due to temperature change, which is a very stable composition.

And, the cosmetic composition according to the present invention exhibits a tyrosinase inhibitory activity, that is, a whitening activity of about 30 to 50%. That is, the cosmetic composition of the present invention retains the activity of the lotus leaf extract even after the cosmetic manufacturing process. Therefore, the cosmetic composition of the present invention will also exhibit an anti-aging effect which is one of the activities of the lotus leaf extract.

The invention can be better understood by the following examples, which are only illustrative of the invention and are not intended to limit the scope of protection defined by the appended claims.

Example 1: Preparation of lotus leaf extract using an enzyme

The lotus leaf was prepared, dried, and ground to a size of 100 mesh. Then, after adding distilled water corresponding to 20 times the volume of the lotus leaf, distilled water containing the lotus leaf was adjusted to pH 5-8 using 1M HCl and 0.1N NaOH. A pen tryptophan ^TM (PENTOPN), serenity mix ^TM (CEREMIX) decomposing enzyme to said pH is adjusted mixture solution, Hatori atom ^TM (VISCOZYME), Ultra Flow ^TM (ULTRAFLO), pekti Annex ^TM (PECTINEX), cellulose Cloud host ^TM (CELLUCLAST), is added in Tunisia Carr claim ^TM (TUNICASE), promotional atom ^TM (PROMOZYME) 8 jong respectively 2% (w / w) of the enzyme. Each sample was placed in a constant temperature water bath and subjected to enzymatic treatment at 50 ° C. for 4 hours.

After the enzyme treatment, each sample was filtered with Whatman No. 41 to obtain a lotus leaf extract.

Experimental Example 1 Measurement of Polyphenol Content, Antioxidant Effect and Tyrosinase Inhibitory Effect of Lotus Leaf Extract According to Kinds or Concentrations of Degrading Enzyme]

Experimental Example 1-1: Measurement of Total Polyphenol Content of Lotus Leaf Extract for Yield Confirmation

The total polyphenol content of the extract obtained in Example 1 was measured using the Folin-Denis assay method. First, 50 ml of Folin-ciocalteu reagent (phosphomolybdate-phosphotungstate) was added to 0.1 ml of the lotus leaf extract obtained in Example 1, followed by reaction with 1.5 ml of 20% sodium carbonate anhydrous, followed by absorbance at 760 nm. Measured. The polyphenol content of the lotus leaf extract according to the type of the degrading enzyme measured is shown in Table 1 below.

Table 1: Polyphenol Contents of Lotus Leaf Extract According to Degrading Enzymes

Enzyme Treatment Condition Total Polyphenol Content (µg / mL) Yield (%) compared to no enzyme treatment Enzyme-free treatment 1176.6 ± 36.5 100 Pentophan 500 BG 1092.5 ± 54.4 92.8 (-7.2) Ceremix 6X MG 1409.9 ± 96.2 119.7 (+19.7) Biscozyme 1151.1 ± 14.6 97.8 (-2.2) Ultraflow L 1214 ± 59.7 103.2 (-3.2) Pectinex 100 L 1079.2 ± 30.9 91.7 (-8.3) Cellulast 1323.7 ± 74.1 112.5 (+12.5) Tunicase FN 1135.1 ± 34.5 96.5 (-3.5) Promozyme 1476.6 ± 60.2 125.5 (25.5)

In Table 1 above,

* Tryptophan pen 500 BG ^TM (Pentopan 500 BG) is a single enzyme of claim xylene Lana (Xylanase).

* Ceremix 6X MG ^TM (Ceremix 6X MG) is a cellulase, proteinase, alpha-amylase, and pentosanase in beta-glucanase. It is an enzyme which mixed four types of enzymes, such as each by the same weight ratio.

* Viscozyme ^TM (Viscozyme) is a beta-glucanase (β-glucanase) cellulase (cellulase), hemicellulase (hemicellulase), xylanase (arabanase) (arabanase) It is an enzyme which mixed enzyme in the same weight ratio, respectively.

4, such as cellulase (Cellulase), xylene Rana claim (Xylanase), pentosan xylanase (pentosanase), Ara bar or the (arabanase) the glucanase the (β-glucanase) - * ultra flow L ^TM (Ultraflo L) is a beta It is an enzyme which mixed the enzyme of a species in the same weight ratio, respectively.

* Pectinex 100L ^TM (pectinex 100L) is a polygalacturonase, pectinesterase, cellulase, hemicellulase, etc. in pectintranseliminase. It is an enzyme which mixed four types of enzyme in the same weight ratio, respectively.

Cellullast ^TM is a cellulase homoenzyme.

Tunicase FN ^™ is a beta-glucanase homoenzyme.

Promozyme ^TM is a Pullulanase homoenzyme.

As shown in Table 1, the polyphenol content of the lotus leaf extract when treated with the addition of Ceremix 6X MG ^TM (Ceremix 6X MG), Cellullast ^TM (Celluclast), Promozyme ^TM (Promozyme) It was 12 ~ 25.5% higher than the enzyme-free group.

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Experimental Example 1-2: Antioxidant Activity of Lotus Leaf Extract According to Kinds of Degrading Enzymes

DPPH radical scavenging ability (%) of the lotus leaf extract obtained in Example 1 was measured to confirm the antioxidant activity of the lotus leaf extract. At this time, Vitamine E was used as a positive control group. As a result of the experiment, all lotus leaf extracts showed more than 70% antioxidant activity except for treatment with Biscozyme ^TM (Viscozyme L) as shown in Table 2 below.

Table 2: Antioxidant Activity of Lotus Leaf Extracts by Kinds of Degrading Enzymes]

Enzyme Treatment Condition Antioxidant activity (%) Enzyme-free treatment 71.6 ± 0.86 Pentophan 500 BG 77.8 ± 1.1 Ceremix 6X MG 83.3 ± 2.7 Biscozyme 67.4 ± 1.3 Ultraflow L 82.9 ± 2.4 Pectinex 100 L 83.1 ± 1.2 Cellulast 80.7 ± 0.49 Tunicase FN 75.6 ± 2.0 Promozyme 83.9 ± 1.4

As shown in Table 2, comparing the enzyme-specific DPPH radical scavenging ability (%) showed higher antioxidant activity than other enzymes in Promozyme ^TM (Promozyme) and Ceremix 6X MG ^TM (Ceremix 6X MG).

Experimental Example 1-3: Antioxidant Activity of Lotus Leaf Extract According to Degrading Enzyme Concentration
Distilled water corresponding to 20-fold was added to the lotus leaf crushed into 100 mesh, and the pH was adjusted to 5 using 0.1N NaOH and 0.1N HCl. Here, 0% (no addition), 0.5%, of Ceremix 6X MG ^TM (Ceremix 6X MG) and Promozyme ^TM (Promozyme), which are the degrading enzymes with the highest polyphenol content of the lotus leaf extract in Experimental Example 1-1, respectively 1%, 2%, 4% (w / w) was added, put in a constant temperature water bath and then enzymatically treated at 50 ℃ for 4 hours. And each lotus leaf extract was obtained by the same method as Example 1.

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At this time, the antioxidant activity (DPPH radical scavenging ability) of the lotus leaf extract according to the concentration of the enzyme is shown in Table 3 below.

Table 3: Antioxidant Activity of Lotus Leaf Extracts According to Degrading Enzyme Concentration

Enzyme concentration 0 0.5 One 2 4 Ceremix 6X MG 81.5 ± 0.87
75 ± 1.4 84.9 ± 0.7 84.3 ± 2.2 84.6 ± 2.0 Promozyme 83.2 ± 0.4 84 ± 0.5 81.5 ± 1.8 80.9 ± 2.0

As shown in Table 3, the lotus leaf extract obtained by treating Ceremix 6X MG ^TM and Promozyme ^TM at a concentration of 1% or 2% (w / w), respectively, was higher than the lotus leaf extract extracted without enzyme treatment. It showed antioxidant activity.

Experimental Example 1-4: Inhibition of Tyrosinase Activity of Lotus Leaf Extract According to Kinds of Degrading Enzymes

The inhibitory effect of Tyrosinase activity of the lotus leaf extract obtained in Example 1 was measured. At this time, Kojic acid was used as a positive control group.

The results of measuring the inhibitory effect of Tyrosinase activity are shown in Table 5 below.

[Table 4: Tyrosinase Activity Inhibitory Effects of Lotus Leaf Extracts According to the Types of Degrading Enzymes]

Enzyme Treatment Condition Whitening activity (%) Enzyme-free treatment 47 ± 9.5 Pentophan 500 BG 55.2 ± 2.7 Ceremix 6X MG 56.7 ± 2.1 Biscozyme 59.7 ± 9.7 Ultraflow L 62.9 ± 0.6 Pectinex 100 L 63.2 ± 3.3 Cellulast 65.2 ± 3 Tunicase FN 64.2 ± 1.8 Promozyme 52.4 ± 9

As shown in Table 4, the best inhibitory effect of Tyrosinase activity was the lotus leaf extract obtained by cellulose treatment, and all lotus leaf extracts obtained by treating each enzyme including Ceremix 6X MG or Promozyme were not treated with enzyme. It showed much higher Tyrosinase inhibitory activity.

Experimental Example 1-5: Inhibition of Tyrosinase Activity of Lotus Leaf Extract According to Degrading Enzyme Concentration

Each lotus leaf was treated with Ceremix 6X MG ^TM and Promozyme ^TM at 0% (no addition), 0.5%, 1%, 2%, 4% (w / w) in the same manner as in Experimental Examples 1-3. An extract was obtained.

At this time, Tyrosinase activity inhibitory effect of the lotus leaf extract according to the concentration of the enzyme is shown in Table 5 below.

Table 5: Inhibitory Effect of Lotus Leaf Extract on Tyrosinase Activity According to Degrading Enzyme Concentration

Enzyme concentration 0 0.5 One 2 4 Ceremix 6X MG 47 ± 7.2
56.5 ± 5.8 53.5 ± 4 76.2 ± 5 59.9 ± 2.7 Promozyme 57.6 ± 4.7 45.9 ± 3 64.5 ± 4.1 55.1 ± 2.7

As shown in Table 5, as a result of measuring the inhibitory effect of Tyrosinase activity of the enzyme-treated extract for each concentration, both Ceremix 6X MG and promozyme showed a high activity inhibitory effect at a concentration of 2% (w / w).

As a result, the polyphenol content and Tyrosinase activity inhibitory effect of each lotus leaf extract showed excellent effects when treated with Ceremix 6X MG or promozyme, and when treated with an enzyme concentration of 2% (w / w) Was the best.

Experimental Example 2: Whitening Activity and Antioxidant Activity of Lotus Leaf Extract Obtained by Promozyme Treatment

In Experimental Example 1, the lotus leaf extract obtained by treating the promozyme among the degrading enzymes was found to have the highest polyphenol content and an excellent inhibitory effect on tyrosinase activity. In Experimental Example 2, the whitening and antioxidant activity of the lotus leaf extract obtained by treating the promoter was examined in more detail.

Experimental Example 2-1: Determination of Tyrosinase Activity Inhibition Effect

Tyrosinase inhibitory activity was added to 0.5 ml of 0.5 M phosphate buffer solution (pH 6.5) and 0.2 ml of substrate solution in which 10 mM L-DOPA was dissolved, and the lotus leaf extract obtained in the same manner as in Example 1 (decomposition enzyme was used as promozyme). 0.2 ml of mushroom tyrosinase (100 U / ml) was added to the mixed solution by adding 0.1 ml of the mixture, and reacted at 25 ° C. for 2 minutes, and then the dopachrome produced in the reaction solution was enzyme Absorbance was measured and compared at 475 nm using a spectroscopic absorbance analyzer (microplate reader).

Figure 2a is a graph showing the tyrosinase inhibitory activity of kojic acid (kojic acid) used as a control drug, Figure 2b is a graph showing the tyrosinase inhibitory activity of the lotus leaf extract. 2A and 2B, as the concentration of kojic acid increased, tyrosinase inhibitory activity was significantly increased to about 70% to about 90%, and the lotus leaf extract inhibited tyrosinase as the concentration increased. Activity was significantly increased by about 60%.

Experimental Example 2-2: Melanin Production Inhibitory Effect

The cell line used in the experiment was purchased from Korean Cell Line Bank (KCLB) as B16F1, and the B16F1 cells were inoculated in DMEM medium and then cultured at 37 ° C. and 5% CO ₂ . Cells attached to the culture dish were passaged with trypsin-EDTA.

The initial concentration of the cells was adjusted to 1 × 10 ⁶ cells / ml, and 200 μl was dispensed into 96 well plates, incubated for 24 hours, and then the medium was removed. The lotus leaf extract obtained in the same manner as in Example 1 (degrading enzyme was used as a promozyme) ) Was incubated for 72 hours. After removal of the culture solution, washed with PBS, 50 μl of 1N NaOH was added, reacted for 2 hours, dissolved, and the absorbance was measured at 400 nm.

3 is a graph showing the melanin production inhibitory ability of the lotus leaf extract according to the present invention. As shown in FIG. 3, it was found that the higher the concentration of the lotus leaf extract, the higher the melamine production inhibitory ability. In addition, when the amount of the lotus leaf extract was 720 µg / ml, it was found that the melamine production inhibitory ability was excellent to the extent that the melamine production inhibitory activity was nearly 100%.

Experimental Example 2-3: Antioxidant Activity-DPPH Radical Scavenging Activity

160 μl of DPPH solution and 40 μl of sample dissolved in methanol at 0.4 mM were added thereto, and the mixture was left in the dark for 30 minutes, and then absorbance was measured at 515 nm using an enzyme spectroscopic absorber (microplate reader). DPPH radical scavenging ability was expressed as a decrease in absorbance of the addition and non-addition of the lotus leaf extract obtained by the same method as in Example 1 as shown in the following inhibition rate (%). Ascorbic acid and alpha tocopherol were used as controls.

% Inhibition = (absorbance of 1-sample addition / absorption of no addition) × 100

Figure 4a is a graph showing the DPPH radical scavenging ability of alpha tocopherol used as a control drug, Figure 4b is a graph showing the DPPH radical scavenging ability of the lotus leaf extract according to an embodiment of the present invention. As shown in FIG. 4A and FIG. 4B, as the concentration of alpha tocopherol was increased, DPPH radical scavenging activity was increased to about 100%, and lotus leaf extract also increased DPPH radical scavenging activity to about 76% as the concentration was increased. It was high.

Experimental Example 2-4: Measurement of SOD (Superoxide Dismutase) -like Activity

The SOD-like activity was 40 μl in 120 μl Tris-HCl buffer (pH 8.5) (the lotus leaf extract obtained in the same manner as in Example 1 (degrading enzyme used promozyme)) and 40 μl 7.2 mM pyrogallol. The reaction was carried out at 25 ° C. for 10 minutes to measure the absorbance at 420 nm. The degree of antioxidant activity can be assessed to the extent of SOD-like activity.

SOD-like activity (%) = (absorbance of 1-sample addition / absorption of no addition) x 100

Figure 5a is a graph showing the SOD-like activity of ascorbic acid used as a control, Figure 5b is a graph showing the SOD-like activity of the lotus leaf extract according to an embodiment of the present invention. As shown in FIG. 5A and FIG. 5B, as the concentration of ascorbic acid increases, the SOD-like activity increases to about 100%, and the lotus leaf extract according to the present invention also increases the SOD-like activity by about 30%. Exceeded.

As seen through the examples and experimental examples, the lotus leaf extract according to an embodiment of the present invention has a high polyphenol content, a high whitening activity effect such as tyrosinase activity inhibitory effect and melanin production inhibitory activity, DPPH It can be seen that the anti-aging effects such as radical scavenging activity and SOD-like activity are excellent.

Hereinafter, a prototype of the cosmetic composition containing the lotus leaf extract prepared according to Example 1 (using a promozyme as an enzyme) as an active ingredient was prepared.

[Example 2: Prototype Preparation of Cosmetic Composition Containing Lotus Leaf Extract as Active Ingredient]

6A is a flowchart illustrating a method of preparing a lotus leaf extract-containing cleansing cream according to a preferred embodiment of the present invention, and FIG. 6B is a flowchart illustrating a method of preparing a lotus leaf extract-containing foam cleansing according to a preferred embodiment of the present invention.

The cosmetic composition prototype containing the lotus leaf extract prepared according to the manufacturing method of FIGS. 6A and 6B was prepared with the composition shown in Table 4 below.

Table 4: Raw Material Composition of Lotus Leaf Extract-Containing Cosmetic Composition Prototype]

cleansing cream Foam cleansing
Purified water, lotus leaf extract 15%, stearic acid, glycerin, lauric acid, cochymidopropyl betaine, potassium hydroxide, myritic acid, PIG-100 stearate, sodium uriryl selperate, rice extract 1% , Polysorbate 20, Fragrance, Propylparaben, Methylparaben

Mineral oil, purified water, lotus leaf extract 14%, propylene glycol, squalane, isopropyl myristate, glyceryl stearate, polysorbate 60, dimethicone, lotus extract (western yellow flower extract) 1%, rice extract 1%, stearic acid, beeswax, cetyl alcohol, petrolatum, PIG-100 stearate, PIG-60 hydrogenated gester oil, triethanolamine, carbomer, methylparaben, fragrance, aloe vera leaf powder, Allatoin, Propylparaben, Disodium IDase, Butylparaben

Experimental Example 4: Measurement of the Stability of the Lotus Leaf Extract-Containing Cosmetic Composition Prototype

Experimental Example 4-1: Stability Test According to Temperature

The cosmetic composition prototype prepared according to an embodiment of the present invention was stored at a predetermined temperature (25 ° C., 45 ° C., 65 ° C.), and the state change according to the change over time was evaluated. The pH was measured at 25 ° C. Before the measurement, the glass electrode was immersed in basic buffer or distilled water in advance, and the pH meter was used after being connected to a power source for at least 10 minutes. The detector was washed well with distilled water and gently wiped. Viscosity measurement was performed using a Brookfield viscometer and measured the viscosity at spindle 3 at 1 rpm for 3 minutes.

Experimental Example 4-1-1: pH Measurement According to Temperature Change

In general, the pH change should be small and stable. In order to determine the pH change with temperature, observation was carried out for 12 days at room temperature, 45 ℃, under the harsh conditions of 65 ℃.

Figure 7a is a graph showing the pH change according to the temperature change of the cleansing cream prepared according to an embodiment of the present invention, Figure 7b is a change in pH according to the temperature change of the foam cleansing prepared according to an embodiment of the present invention The graph shown. As shown in FIG. 7a and FIG. 7b, both the cleansing cream and the foam cleansing prepared according to the method of Example 2 had no significant changes to pH 6.0 ± 0.1 and pH 8.5 ± 0.2, and showed no significant difference and was relatively stable. It was confirmed.

Experimental Example 4-1-2: Viscosity Measurement with Temperature Change

Viscosity is the internal frictional force on both sides of the plane parallel to the flow as the liquid moves in a constant direction, and the viscosity value depends on the temperature and the type of fluid. In addition, the viscosity value may vary depending on the spindle, speed, time, bubbles, and the like.

In this experiment, the sample was fixed at 25 ° C. to minimize the factors that could affect the measurement. As a result of the viscosity measurement, the cleansing cream showed 142,000 cP ± 2000 and the foam cleansing showed a high viscosity of 112,000 cP ± 1000.

Figure 8a is a graph showing the viscosity change according to the temperature change of the cleansing cream prepared according to an embodiment of the present invention, Figure 8b is a viscosity change according to the temperature change of the foam cleansing prepared according to an embodiment of the present invention The graph shown. As shown in FIG. 8A and FIG. 8B, as the temperature increases, the change in foam cleansing is higher than the change in cleansing cream, and the viscosity decreases as time passes.

Experimental Example 4-2: Natural Light Exposure Test

Containing the cosmetic composition prepared according to one embodiment of the present invention in a transparent container in consideration of the conditions under the midsummer sun, discoloration, discoloration, viscosity in the same manner as in Experiment 4-1 for 0 to 12 days outdoors in sunny sunlight Changes in pH, etc. were observed and measured.

Figure 9a is a graph showing the pH change according to natural light exposure of the lotus leaf extract containing cosmetic composition prepared according to an embodiment of the present invention, Figure 9b is a natural light of the lotus leaf extract containing cosmetic composition prepared according to an embodiment of the present invention It is a graph showing the change in viscosity with exposure.

As shown in FIG. 9A and FIG. 9B, the pH change was not significantly different between the cleansing cream and the foam cleansing as a result of storing and observing in a sunny place considering the conditions under the midsummer sun. There was no change in the back. In addition, the viscosity was shown to decrease slightly over time, but the viscosity change during 12 days did not change significantly was confirmed that the lotus leaf extract-containing cosmetic composition according to the present invention is stable.

Experimental Example 5: Whitening Activity of Cosmetic Composition Containing Lotus Leaf Extract

Tyrosinase plays a very important role in skin melanin production, and DOPA oxidase, which produces DOPA quinones by oxidizing DOPA with tyrosine hydroxylase, which oxidizes tyrosine in melanosomes to produce DOPA. oxidase) acts as an important enzyme in the synthesis of melanin polymers.

The preparation composition of the lotus leaf extract-containing cosmetic composition at a concentration of 10% (w / w), the results of measuring the tyrosinase inhibitory activity, that is, the whitening activity of the cosmetic composition is shown in FIG. As shown in FIG. 10, the lotus leaf extract-containing cleansing cream according to an embodiment of the present invention has a whitening activity of 51.1%, the lotus leaf extract-containing foam cleansing shows a whitening activity of 30%, which is lower than that of the lotus leaf extract. However, it was found that the active ingredient activity of the lotus leaf extract remained after tyrosinase inhibitory activity remained even after the cosmetic composition manufacturing process.

According to the above, it can be seen that the cosmetic composition of the present invention containing the lotus leaf extract prepared according to the present invention as an active ingredient retains the whitening activity of the lotus leaf extract even after the cosmetic manufacturing process. Since the active ingredient activity of the lotus leaf extract remains after the cosmetic manufacturing process, it can be seen that the lotus leaf extract containing cosmetic composition according to the present invention also retains the anti-aging effect of the lotus leaf extract.

1 is a flow chart showing a lotus leaf extract manufacturing method according to an embodiment of the present invention.

Figure 2a is a graph showing the tyrosinase inhibitory activity of kojic acid used as a control drug, Figure 2b is a graph showing the tyrosinase inhibitory activity of the lotus leaf extract prepared according to an embodiment of the present invention to be.

3 is a graph showing the melanin production inhibitory ability of the lotus leaf extract prepared according to an embodiment of the present invention.

Figure 4a is a graph showing the DPPH radical scavenging ability of alpha tocopherol used as a control drug, Figure 4b is a graph showing the DPPH radical scavenging ability of the lotus leaf extract prepared according to an embodiment of the present invention.

Figure 5a is a graph showing the SOD-like activity of ascorbic acid used as a control, Figure 5b is a graph showing the SOD-like activity of the lotus leaf extract prepared according to an embodiment of the present invention.

6A is a flowchart illustrating a method of preparing a lotus leaf extract-containing cleansing cream according to a preferred embodiment of the present invention, and FIG. 6B is a flowchart illustrating a method of preparing a lotus leaf extract-containing foam cleansing according to a preferred embodiment of the present invention.

Figure 7a is a graph showing the pH change according to the temperature change of the cleansing cream prepared according to an embodiment of the present invention, Figure 7b is a change in pH according to the temperature change of the foam cleansing prepared according to an embodiment of the present invention The graph shown.

Figure 8a is a graph showing the viscosity change according to the temperature change of the cleansing cream prepared according to an embodiment of the present invention, Figure 8b is a viscosity change according to the temperature change of the foam cleansing prepared according to an embodiment of the present invention The graph shown.

Figure 9a is a graph showing the pH change according to natural light exposure of the lotus leaf extract containing cosmetic composition prepared according to an embodiment of the present invention, Figure 9b is a natural light of the lotus leaf extract containing cosmetic composition prepared according to an embodiment of the present invention It is a graph showing the change in viscosity with exposure.

10 is a graph showing the results of measuring the whitening activity of the lotus leaf extract-containing cosmetic composition prepared according to an embodiment of the present invention.

Claims (6)

delete Preparing a Nelumbo nucifera Gaertner leaf; Adding distilled water to the prepared lotus leaf and adjusting the pH to 5-8; After the pH adjustment, enzymatic treatment by adding cell wall degrading enzyme; And After the enzyme treatment, the step of filtration or centrifugation to obtain a lotus leaf extract, The degrading enzyme is an enzyme mixed with beta-glucanase, cellulase, proteinase, alpha-amylase and pentosanase, The enzyme treatment is lotus leaf extract production method characterized in that the immersion for 1 hour to 4 hours at a temperature of 30 ~ 70 ℃. Preparing a Nelumbo nucifera Gaertner leaf; Adding distilled water to the prepared lotus leaf and adjusting the pH to 5-8; After the pH adjustment, enzymatic treatment by adding cell wall degrading enzyme; And After the enzyme treatment, the step of filtration or centrifugation to obtain a lotus leaf extract, The degrading enzyme is pullulanase, The enzyme treatment is lotus leaf extract production method characterized in that the immersion for 1 hour to 4 hours at a temperature of 30 ~ 70 ℃. delete A lotus leaf extract prepared according to the method of claim 2 or 3, wherein the polyphenol content is 1,100-1,500 µg / ml. Whitening, anti-aging cosmetic composition containing the lotus leaf extract of claim 5 as an active ingredient.

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