TWI792573B - Composition containing croton lechleri resin extract and application thereof - Google Patents

Abstract

The present invention relates to a composition containing croton lechleri resin extract and its application. The croton lechleri resin extract has anti-oxidation and anti-aging effects. In order to enhance its effect, it is combined with other substances that also have anti-oxidation and anti-aging effects to further enhance the anti-oxidation and anti-aging effects.

Description

Compositions comprising peruvian croton resin extract and uses thereof

The present invention relates to a composition containing Peruvian croton resin extract and its application, specifically, to a composition containing Peruvian croton resin extract for anti-oxidation and anti-aging.

With the aging of the world, the skin will gradually weaken with age, so when the skin's antioxidant power is not enough, the oxidation will be out of balance, and the aging phenomenon will be accelerated. The main reason for the increase of this type of active oxygen is called "oxidative stress".

External oxidative stress, such as stimulation caused by ultraviolet rays, air pollution, excessive friction and improper medical treatment, as well as lack of nutrients, unbalanced diet, excessive intake of high-sugar and high-salt food, smoking, and alcoholism. Influence. Intrinsic oxidative stress, for example, can be fatigue, insomnia, staying up late, depression, anxiety, and sleep deprivation, etc., as well as mental stress from the workplace and interpersonal relationships.

Croton Lechleri Resin Extract is a thousand-year-old dracaena in South America. It grows in the Amazon rainforest of Peru. It has large spherical branches and leaves. The off-white trunk can secrete a magical wine-red juice. Local residents call it " Dragon's blood" because of its curative blood red color The juice is like precious dragon blood; the Peruvian croton resin extract has been used by Amazon residents as a magic weapon for curing diseases and anti-aging and rejuvenation since ancient times because of its excellent repairing ability.

Many studies have reported that the Peruvian croton tree has the following effects:

1. Anti-aging effect: Proanthocyanidins contained in Peruvian croton resin can effectively rebuild collagen fiber bundles, restore skin elasticity and toughness, and have anti-aging effects.

2. Wound repairing effect: Lignin in Peruvian croton resin can regulate the mechanism of wound healing, and polyphenols can scavenge free radicals, so it can stimulate wound contraction, stimulate collagen production, and help the movement of fibroblasts Scar formation, accelerate skin renewal, achieve the effect of wound repair.

3. Anti-free radical ability: The proanthocyanidins and catechins in the Peruvian croton resin resin have anti-oxidation effects, which can effectively delay the oxidation of the skin.

From the above content, it can be seen that Peruvian croton resin has anti-oxidation and anti-aging effects; but in order to enhance and optimize the antioxidant and anti-aging effects of Peruvian croton resin extract for skin repair, there is no relevant research on Peruvian croton resin. The composition and ratio of the extract and other ingredients can enhance the antioxidant and anti-aging effects of the Peruvian croton resin extract.

Therefore, it is necessary to develop a composition comprising the Peruvian croton resin extract on the premise of enhancing the antioxidant and anti-aging effects of the Peruvian croton resin extract.

In view of the above-mentioned problems, the object of the present invention is to overcome the shortcomings of the above-mentioned prior art, and further provide a composition containing Peruvian croton resin extract, which has better anti-oxidation and anti-aging effects.

The composition comprising Peruvian croton resin extract provided by the present invention comprises Peruvian croton resin extract, clary sage essential oil, tea tree essential oil, tocopherol, hydrogenated castor oil, 1,3-propanediol and water.

Among them, clary sage essential oil is extracted from clary sage, which is a plant of the genus Salvia (Lamiaceae) and the main component of the essential oil extracted from it is linalool (Linalool), Linalyl acetate, α-terpineol, Germacrene D, Geranyl acetate, and Sclareol; These ingredients also have antioxidant effects.

Tea tree essential oil is extracted from the branches and leaves of tea tree. Tea tree is a plant of the genus Melaleuca in the family Myrtaceae. The main components of the essential oil extracted from it are 4-terpineol (Terpinen-4-ol). ), γ-terpinene (γ-terpinene), 4-carene (4-carene), α-terpineol (α-terpineol) and terpinolene (Terpinolene), of which γ-terpinene is in the body It has good anti-oxidation function inside and outside, effectively combines free radicals and reactive oxygen species inside and outside the cell membrane, significantly increases superoxide dismutase (SOD), reduces oxidative damage, and maintains cell activity.

The remaining ingredients in the composition, such as tocopherol and hydrogenated castor oil, are common compounds with antioxidant effects, while 1,3-propanediol is used as a common solvent, moisturizer or thickener in cosmetics and skin care products.

In addition, in the composition provided by the present invention comprising Peruvian croton resin extract, each component is based on the total weight of the composition, and its weight percentage is shown in Table 1 below, wherein the "remainder" refers to On a weight basis, after the content of each component is determined, the sum of the added 1,3-propanediol amount and the content of other components is just equal to the total weight of the composition.

Furthermore, the composition comprising the Peruvian croton resin extract provided by the present invention can be obtained only by mixing the ingredients uniformly according to the proportion; therefore, the preparation method is simple and the manufacturing cost is cheap.

To sum up, by mixing the Peruvian croton resin extract with other ingredients of specific types and ratios, a composition with significant anti-oxidation and anti-aging effects can be obtained.

The following diagrams are used to further illustrate the present invention: Figure 1 is a schematic diagram of the DPPH free radical scavenging ability of only the Peruvian croton resin extract; Figure 2 is a combination of different composition ratios containing the Peruvian croton resin extract The schematic diagram of the DPPH free radical scavenging ability of the croton resin extract and only the Peruvian croton resin extract; Figure 3 is a schematic diagram of the total phenol content of the composition containing the Peruvian croton resin extract and only the Peruvian croton resin extract in different composition ratios ; Figure 4 is a schematic diagram of the use of different composition ratios and concentrations of the composition comprising the Peruvian croton resin extract and only the first collagen produced by the Peruvian croton resin extract; Fig. 5 is a schematic diagram of the effect on cell viability of using different composition ratios and concentrations of compositions containing Peruvian croton resin extract and only Peruvian croton resin extract.

The following content will be combined with drawings to illustrate the technical content of the present invention through specific embodiments, and those who are familiar with this technology can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments. Various modifications and changes may be made to the details in this specification based on different viewpoints and applications without departing from the spirit of the present invention.

Experimental Materials

Composition containing Peruvian croton resin extract, consisting of the following 6 ingredients and water: Peruvian croton resin extract (Croton Lechleri Resin Extract, Cobiosa, Spain), clary sage oil (Clary Sage Oil, Payan Bertrand, France), tea tree essential oil ((Melaleuca Alternifolia (Tea Tree) Leaf Oil, Southern Cross Botanicals, Australia), tocopherol (Tocopherol, Fuji Chemical Industry, Japan), hydrogenated castor oil (PEG-60 Hydrogenated Castor Oil, BASF, Germany) , 1,3-propanediol (Propanediol, DuPont Tate & Lyle Bio Products, USA).

experimental method

1. DPPH free radical scavenging test

In the process of lipid self-oxidation, free radicals will be generated to cause lipid rancidity. Common antioxidants provide hydrogen (Hydrogen doner) to scavenge lipid peroxide free radicals (Peroxyl freeradical), thereby inhibiting the oxidation chain reaction. conduct.

DPPH (2,2-Diphenyl-1-picrylhydrazyl) is a stable free radical, and it has a strong absorbance value at 517nm with ethanol (Methanol) solution, so the lower the absorbance value, the hydrogen supply capacity of antioxidants The stronger, the better the antioxidant capacity.

DPPH free radical scavenging rate (%)=(1-A/B)×100%

Among them, A represents the absorbance value of the sample group at OD517, and B represents the absorbance value of the control group at OD517.

First, take 40 μL of samples from the comparison group and the experimental group diluted with different concentrations, and add them to different holes in the 96-well cell analysis plate, and then add 160 μL of 0.3 mM DPPH ethanol solution, mix evenly, and then stand in the dark at room temperature After reacting for 30 minutes, the absorbance value (OD517) at a wavelength of 517nm was measured with a Microplate reader (SPECTROstar Nano), wherein the concentrations of the samples in the experimental groups were 0.625%, 1.25%, 2.5%, 5%, and 10%.

2. Determination and analysis of total phenol content

Use phenolic indicator (Folin-Ciocalteu's phenol reagent) to detect the content of phenolic compounds in the sample; if there are phenolic compounds in the sample, it will react with the phenolic indicator at a wavelength of 750nm, and when the absorbance value is higher, it means The more polyphenols contained in the sample, the stronger the antioxidant capacity. Prepare different known concentrations of gallic acid (gallic acid) and react with phenolic indicators as a standard product and make a calibration line to detect the absorption value of the unknown substance after reacting with the indicator, and the corresponding calibration line can be measured content of phenolic compounds.

First, take 200 μL of samples from the comparison group and the experimental group at different concentrations into different microcentrifuge tubes, and add 100 μL of sodium carbonate (Na ₂ CO ₃ ) and 100 μL of phosphomolybdic acid phenol reagent to each microcentrifuge tube, React at room temperature for 90 minutes in the dark. Afterwards, 200 μL of the reaction solution was taken in each microcentrifuge tube to different holes in the 96-well cell analysis plate, and the absorbance value (OD750) at a wavelength of 750 nm was measured with a Microplate reader (SPECTROstar Nano), and each experimental group was carried out three times. The test was repeated, wherein the final concentration of the composition of the comparison group sample and the experimental group was 6.25%. In this determination analysis, gallic acid was also used as a positive control group to compare the relative total phenolic content of the composition of the present invention compared with gallic acid (positive control group).

3. Procollagen type I production test

Type I collagen is the most abundant collagen in the human body. It forms large eosinophilic fibers called collagen fibers, promotes the synthesis of type I collagen and the generation of fibroblasts in the skin, and has anti-aging and anti-wrinkle effects , It can repair and maintain skin elasticity in the skin function.

Hs68 cells were seeded in a 96-well cell assay plate, and the number of cells per well was 1x10 ⁵ . After culturing in a cell incubator at 37°C/5% CO ₂ for 24 hours, remove the original cell culture medium, and add the diluted samples (each sample was diluted to 0.25%, 0.5% and 1%) into the analysis plate , and each sample was tested in triplicate. After culturing for 72 hours, the supernatant was transferred to an EIA Kit assay plate (Precoated) for enzyme immunoassay (ELISA).

4. Cell Viability Analysis

(formazan)結晶而分辨細胞是否存活，由於死亡細胞之粒線體中不具有活性之脫氫酶，所以MTT的顏色仍然為黃色不會產生藍紫色之甲

結晶。而甲

結晶於波長570nm時具有最大吸光值，因此可經由吸光值的測定來計算細胞的存活率。 The analysis of cell viability is based on the ability of succinate dehydrogenase in the mitochondria of Hs68 cells to convert MTT ([3-(4,5)-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) into a light yellow aqueous solution The tetrazole ring is cut off to produce blue-purple carapace

(formazan) crystallization to distinguish whether the cells are alive or not, because the mitochondria of dead cells do not have active dehydrogenase, so the color of MTT is still yellow and will not produce blue-purple formazan

crystallization. And A

The crystal has a maximum absorbance value at a wavelength of 570nm, so the cell survival rate can be calculated by measuring the absorbance value.

Cell survival rate formula (%)=A/B×100%

Among them, A represents the absorbance value of the sample group at OD570, and B represents the absorbance value of the control group at OD570.

Experimental results

Example 1

Analysis of DPPH free radical scavenging test of Peruvian croton resin extract

Dissolve the Peruvian croton resin extract in a mixture of water and 1,3-propylene glycol, and prepare 0.1%, 0.5% and 1% by weight compositions containing only the Peruvian croton resin extract as comparison groups RA and RB And RC, its mixing ratio is shown in Table 2 below:

First, carry out DPPH test with the above-mentioned comparison groups, and preliminarily select the appropriate ratio of Peruvian croton resin extract to use, and calculate IC50 from the experimental results. IC50 is the concentration required to inhibit 50% of free radicals. The lower the concentration, the more antioxidant. The stronger the force.

Referring to Figure 1, Figure 1 is a schematic diagram of the DPPH free radical scavenging ability of only Peruvian croton resin extract. As shown in Figure 1, when the IC50 is reached, the comparison group R-A is 42.40wt%, R-B is 8.15wt%, and R-C is 8.13wt%. It can be seen that when the content of Peruvian croton resin extract increases from 0.1wt% to 0.5 The concentration when it reached IC50 dropped significantly from 42.4wt% to 8.15wt% at wt%, but when the content of Peruvian croton resin extract increased from 0.5wt% to 1.0wt%, the concentration when it reached IC50 was not clear Therefore, based on R-B as the best anti-oxidation comparison group (that is, the use of Peruvian croton resin extract is 0.5%), and adding other ingredients for subsequent anti-oxidation and anti-aging tests.

Example 2

Test Analysis of DPPH Free Radical Scavenging Ability of Compositions Containing Peruvian Croton Resin Extract

Based on the comparison group RB (that is, the amount of Peruvian croton resin extract used is 0.5%), adding appropriate proportions of clary sage essential oil, tea tree essential oil, tocopherol, hydrogenated castor oil, 1,3-propanediol and water as the experimental group AD, after uniformly mixing the Peruvian croton resin extract with the above-mentioned ingredients, a composition containing the Peruvian croton resin extract was obtained. The individual mixing ratios of the experimental groups AD are shown in Table 3 below:

The experimental group A-D was subjected to the DPPH test and compared with the comparison group R-B. The DPPH free radical scavenging ability is shown in Figure 2; it can be seen from the second figure that the concentration of the experimental group A-D when reaching IC50 is 5.14 wt%, 6.87wt%, 7.54wt% and 5.26wt% (that is, the concentration when the experimental group A-D reaches IC50 degree range of about 5~8wt%), its DPPH free radical scavenging ability is greater than that of the comparison group R-B (8.15wt%), and in the experimental group A-D, the experimental group A, D has better DPPH than the experimental group B, C Free radical scavenging ability, especially the experimental group A has the strongest DPPH free radical scavenging ability.

Example 3

Determination and Analysis of Total Phenol Content of Compositions Containing Peruvian Croton Resin Extract

Comparison group R-B and experimental group A-D are carried out total phenolic content measurement analysis, and its result is as shown in the 3rd figure; As can be seen from the 3rd figure, experimental group A-D is relative to the total phenolic content of gallic acid (mg of GAE /g, GAE is the abbreviation of gallic acid equivalent (gallic acid equivalent)) are 0.1181 (mg of GAE/g), 0.0323 (mg of GAE/g), 0.0304 (mg of GAE/g) and 0.0997 ( mg of GAE/g), that is, the total phenol content of the experimental group A-D ranged from about 0.03 to 0.12 (mg of GAE/g), while that of the comparison group R-B was only 0.0277 (mg of GAE/g).

From the above results, it can be seen that the total phenolic content of the experimental group A-D is higher than that of the comparative group R-B, and among the experimental groups A-D, the experimental group A and D have higher total phenolic content than the experimental group B and C, especially the experimental group A has the highest total phenol content; this result is consistent with the trend of DPPH free radical scavenging ability in Example 2.

Example 4

Analysis of Type I Procollagen Production Test

The comparison group R-B and the experimental group A-D were subjected to type I collagen production test analysis, and the results are shown in Figure 4; it can be seen from Figure 4 that compared with the control group, the production of type I collagen The comparison group R-B (1%) increased by 146%, the experimental group A (1%) increased by 174%, the experimental group B (1%) increased by 171%, the experimental group C (1%) increased by 104%, and the experimental group D ( 1%) increased by 103%, so it can be seen that the comparison group R-B and the experimental groups A and B have a positive and significant difference in the production of type I collagen at a concentration of 1% (the * in Figure 4 indicates P<0.05, and ** indicates P<0.01).

In addition, when the concentration of each group was increased, the production of type I collagen also increased; the results proved that the production of collagen in the experimental group A and B was better than that of the comparison group R-B, and at 1 % concentration compared with the comparison group R-B sequentially increased by 27%, 25% (increased by 1.18 times and 1.17 times), that is, increased by more than 25%. Therefore, after mixing the Peruvian croton resin extract with the above-mentioned specific ingredients in a specific ratio, it can more effectively increase the production of type 1 collagen and achieve anti-aging effects.

Example 5

Cell Viability Analysis

The comparison group R-B and the experimental group A-D were subjected to type I collagen production test analysis, and the results are shown in Figure 5; it can be seen from Figure 5 that, compared with the control group, except for the experimental group D (1% ) was less than 80%, the cell survival rates of the comparison group R-B and the experimental group A-D with different concentrations were all greater than 80%. Therefore, it can be known that the composition comprising the Peruvian croton resin extract is not very toxic to cells.

In summary, the experimental results in each example confirm that the antioxidant capacity and total phenolic content of the experimental groups A, B, C, and D are better than those of the comparison group R-B, while the collagen production of the experimental groups A and B The amount is better than that of the comparison group R-B. It can be seen that the composition containing the Peruvian croton resin extract in the experimental groups A and B has the best anti-oxidation and anti-aging effects, and is not very toxic to cells, so the composition can be used in cosmetics or skin care products middle.

Those in the technical field of the present invention can understand from the foregoing that the present invention can be exemplified in other specific forms without changing the technical concepts or essential features of the disclosure. In this regard, the exemplary aspects disclosed herein are for illustrative purposes only and should not be construed as limiting the scope of this disclosure. Rather, this disclosure is intended to cover not only such exemplary aspects, but also Various changes, modifications, equivalents, and other aspects within the spirit and scope of the present invention as defined in the appended claims.

Claims (6)

A composition comprising Peruvian croton resin extract, which comprises the following ingredients: Peruvian croton resin extract (Croton Lechleri Resin Extract), clary sage oil (Clary Sage Oil), tea tree essential oil (Melaleuca Alternifolia Leaf Oil), tocopherol (Tocopherol), hydrogenated castor oil (PEG-60 Hydrogenated Castor Oil), 1,3-propanediol (Propanediol) and water, wherein based on the total weight of the composition, the Peruvian croton resin extract is 0.5wt%, clary sage 0.3~0.7wt% of essential oil, 0.2wt% of tea tree essential oil, 0.1~0.5wt% of tocopherol, 0.1~2wt% of hydrogenated castor oil, 4.5wt% of water, and 1,3-propanediol as the balance; and When the DPPH free radical scavenging ability of the composition reaches IC50, the use concentration of the composition is 5~8wt%, and the total phenol content of the composition relative to gallic acid is 0.03~0.12 (mg of GAE/g) . The composition comprising the Peruvian croton resin extract as described in claim 1, wherein the production of the first type collagen (Procollagen type I) of the composition is increased by 25% compared with only the Peruvian croton resin extract above. The composition comprising Peruvian croton resin extract as claimed in claim 1, wherein when the composition is used in Hs68 cells, the survival rate of the Hs68 cells is greater than 80%. A composition comprising Peruvian croton resin extract is used for anti-oxidation and anti-aging applications. The composition comprises the following ingredients: Peruvian croton resin extract (Croton Lechleri Resin Extract), clary sage essential oil (Clary Sage Oil), Tea Tree Essential Oil (Melaleuca Alternifolia Leaf Oil), Tocopherol (Tocopherol), Hydrogenated Castor Oil (PEG-60 Hydrogenated Castor Oil), 1,3- Propanediol (Propanediol) and water, wherein based on the total weight of the composition, Peruvian croton resin extract is 0.5wt%, clary sage essential oil is 0.3~0.7wt%, tea tree essential oil is 0.2wt%, tocopherol is 0.1~ 0.5wt%, hydrogenated castor oil is 0.1~2wt%, water is 4.5wt%, and the balance is 1,3-propanediol; and when the DPPH free radical scavenging ability of the composition reaches IC50, the use concentration of the composition is 5~8wt%, and the total phenol content of the composition relative to gallic acid is 0.03~0.12 (mg of GAE/g). The composition comprising Peruvian croton resin extract as described in claim 4 is used for anti-oxidation and anti-aging applications, wherein when the composition has only Peruvian croton resin extract, its type I collagen (Procollagen type I) generation increased by more than 25%. The composition comprising Peruvian croton resin extract as described in claim 4 is used for anti-oxidation and anti-aging applications, wherein when the composition is used in Hs68 cells, the survival rate of the Hs68 cells is greater than 80%.

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