KR20180088146A - Cosmetic composition for antiaging and whitening derived from lipid-extracted microalgae - Google Patents

Abstract

An object of the present invention is to provide an anti-aging and whitening cosmetic composition derived from defatted microalgae which has little cytotoxicity and is excellent in antioxidant, whitening and anti-aging effects. The present invention provides the cosmetic composition having excellent antioxidative, whitening and anti-aging effects by using the defatted microalgae remaining after an extraction process to produce biodiesel.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for anti-aging and whitening derived from defatted microalgae,

The present invention relates to a cosmetic composition for anti-aging and whitening, and more particularly to a cosmetic composition for anti-aging and whitening derived from a defatted microalgae.

In order to lower the production of fatty acid methyl esters derived from microalgae, securing a by-product credit through the use of a lipid-extracted microalgae, a by-product of lipid extraction . As a result of mass cultivation of microalgae, it is possible to mass-produce microalgae with high lipid content, and at the same time, there is a problem of disposal of micro-fine algae produced as a byproduct. Therefore, in order to recycle microalgae, Butylated Hydroxyanisole, Butylated Hydroxytolune, Hydroquinone, Catechol and DPPD have been widely used for their high economic efficiency with strong antioxidative effect. However, It is a reality that there is a high demand for finding a natural antioxidant that is harmless to the human body and has high antioxidant effect because of increasing rejection and decreasing use. Currently, we have developed functional bioactive materials using natural biological materials that have little harmful effects on humans and the environment. We have developed natural antioxidants such as searching for antioxidants from new plant resources, isolating substances, and finding functional materials, need. On the other hand, the degreasing microalgae are mostly composed of cell walls in addition to the residual lipids, and cell walls are formed by xyloglucan and arabinoxylan, which constitute cellulose and hemicellulose, which are a large amount of glucose, Lignin, which is a combination of various phenolic substances, has a form that surrounds and protects the outside. Thus, phenolic compounds are precursors of lignin that form the cell walls of plants, such as gallic acid, coumaric acid, caffeic acid, cinnamic acid, ferulic acid, acid as a polyphenol with multivalent OH- and it is highly utilized as a natural antioxidant. Among the polyphenols that exhibit antioxidant activity, hydroquinone, which is known to have excellent whitening effect, inhibits the production of melanin pigment, which is a cause of stain. Its whitening effect is also known as 10 to 100 times of vitamin C. Inhibition of tyrosinase activity in the melanogenesis process melanogenesis inhibits melanin pigment formation and deposition by inhibiting the oxidation of tyrosine to DOPA in the early stages of melanin biosynthesis. However, hydroquinone derived from a compound shows toxicity to fibroblasts of normal skin and various side effects such as tissue gallstones and partial discoloration of skin color are observed during long-term use, so that it is prohibited to be used as a raw material for whitening products, Is used only as a medicament for the purpose of. In recent years, there has been a growing interest in whitening substances derived from natural materials, which exhibit a strong whitening effect with little side effects on the human body in terms of development of cosmetics. Physiologically active substances including polyphenols contained in seaweeds and microalgae are known to have strong antioxidant ability and are known to have an inhibitory effect on tyrosinase activity. Therefore, they are useful as cosmetic material replacing conventional hydroquinone as a natural whitening material. High value.

When the polyphenol and the physiologically active substance contained in the cell walls of the degreasing microalgae are appropriately separated, they can be used as whitening and natural antioxidants, and thus they are highly valuable as functional materials for high value-added cosmetics and pharmaceuticals. Existing extraction of physiologically active substances by high temperature and high pressure treatment accelerated oxidation at a high temperature, causing rapid deformation of the physico-chemical properties of the physiologically active substance, resulting in the deterioration of function. In this regard, Korean Patent Registration No. 1410632 discloses a cosmetic composition containing fucoxanthin derived from microalgae and a method for producing the same.

However, in the case of the above-mentioned prior arts, there are many problems in using the drug at an effective concentration or higher in terms of stability, discoloration possibility, etc. due to high energy consumption and low extraction efficiency.

It is an object of the present invention to provide an anti-aging and whitening cosmetic composition derived from a defatted microalga having almost no cytotoxicity and excellent antioxidant, whitening and anti-aging effect, to solve various problems including the above- . However, these problems are exemplary and do not limit the scope of the present invention.

According to one aspect of the present invention, there is provided an anti-aging and whitening cosmetic composition derived from a defatted microalgae, which contains a lipid-extracted microalgae extract as an active ingredient.

According to another aspect of the present invention, A protein concentration step of treating degreasing microalgae with an organic solvent to remove lipids and carbohydrates; A protein elution step of eluting the protein by treating a basic solution of the defatted microalgae in which the protein is concentrated; A protein precipitation step of treating the protein eluted in the protein elution step with an acidic solution to precipitate the protein; And a recovery step of removing the protein precipitated in the protein precipitation step and recovering a supernatant or a solid residue from the remaining defatted microalgae, wherein the crude polyphenol is extracted from the defatted microalgae.

According to another aspect of the present invention, Preparing a degreasing microalgae; There is provided a method for extracting crude polyphenols from degreasing microalgae, which comprises an extraction step of extracting the degreasing microalgae using water, an organic solvent or a mixture thereof or ultrasonic waves.

As described above, according to one embodiment of the present invention, the effect of producing a cosmetic composition having excellent antioxidation, whitening and anti-aging effect can be realized by using the defatted microalgae remaining after the extraction process to produce biodiesel. Of course, the scope of the present invention is not limited by these effects.

FIG. 1 is a graph illustrating the effect of microbial destruction size on the production of antioxidant-polyphenol compositions from defatted microalgae according to an embodiment of the present invention.
FIG. 2 is a graph illustrating the effect of solid-to-liquid ratio on the production of antioxidant-polyphenol compositions from defatted microalgae according to an embodiment of the present invention.
FIG. 3 is a graph illustrating the effect of ethanol concentration on the production of antioxidant-polyphenol compositions from defatted microalgae according to an embodiment of the present invention.
FIG. 4 is a graph illustrating the effect of extraction temperature on the production of antioxidant-polyphenol compositions from defatted microalgae according to an embodiment of the present invention.
FIG. 5 is a graph illustrating the effect of extraction time on the production of antioxidant-polyphenol compositions from defatted microalgae according to an embodiment of the present invention.
FIG. 6 is a graph illustrating the effect of extraction time and concentration on the extraction of weak acid of defatted microalgae polyphenols according to an embodiment of the present invention.
FIG. 7 is a graph illustrating the effect of weak acid concentration and extraction temperature on weak acid extraction of defatted microalgae polyphenols according to an embodiment of the present invention.
FIG. 8 is a graph illustrating the effect of weak acid concentration and extraction temperature on the extraction of weak acid of the degreasing microalgae polyphenols according to an embodiment of the present invention.
9 is a graph illustrating the effect of polyphenol concentration on the degreasing microalgae extraction method according to an embodiment of the present invention.
FIG. 10 is a graph illustrating tyrosinase inhibitory activity for evaluating whitening effect of defatted microalgae and microalgae extracts according to an embodiment of the present invention.

Definition of Terms:

As used herein, the term "microalgae" refers to phytoplankton inhabiting the ocean, and plankton such as cochlearinism, which often causes red tides, is also a microalgae. Microalgae, which focuses on marine bioenergy research, is a species of microalgae that is rich in lipids, that is, oil. The size is about 10 microns (micron, one millionth of a meter) and about one tenth the thickness of the hair.

As used herein, the term "lipid-extracted microalgae" is a solid by-product obtained by extracting lipids after microalgae cultivation. It accounts for 30-50% of the dry weight of microalgae. Most of the lipid-extracted microalgae are composed of cell walls, Is composed of xyloglucan and arabinoxylan, which constitute a large amount of cellulose, glucose and hemicellulose, and a lignin, which is a combination of various phenolic substances, is wrapped around the outside It has a form of protection. Since degreasing microalgae contains natural antioxidants including phenolic compounds, it contains a large amount of functional materials such as polysaccharides and antioxidants which are not eluted in the solvent used for lipid extraction, and thus are highly valuable as high value-added foods, cosmetics and pharmaceutical materials have.

As used herein, "melanin" is a proteinaceous organic pigment formed from amino acids that determines human skin color. Melanin is a protein complex that is produced in the melanocyte present in the basal layer of the skin. It is divided into black and brown eumelanin and yellow and red pheomelanin.

DETAILED DESCRIPTION OF THE INVENTION [

According to one aspect of the present invention, there is provided an anti-aging and whitening cosmetic composition derived from a defatted microalgae, which contains a lipid-extracted microalgae extract as an active ingredient.

In the cosmetic composition for anti-aging and whitening derived from the defatted microalgae, the microalgae may be tetracellis, spirulina, chlorella, or vegetable plaque.

The cosmetic composition of the present invention is not particularly limited in the form of the cosmetic composition of the present invention. For example, the cosmetic composition of the present invention can be used in a variety of forms such as a softening agent, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, And may be formulated into cosmetics such as water, a pack, a powder, a body lotion, a body cream, a body oil, and a body essence, and may be applied to a form absorbed into the skin using a form applied to the skin or a micro needle.

In the cosmetic composition of the present invention, the pharmaceutically acceptable carrier or the cosmetically acceptable carrier may include hydrocarbons such as vaseline, liquid paraffin, gelling hydrocarbon (aka plastid base) and the like; Vegetable oils such as medium-chain fatty acid triglyceride, lard, hard fat, cacao butter and the like; Higher fatty alcohols such as cetanol, stearyl alcohol, stearic acid and isopropyl palmitate; fatty acids and esters thereof; Water-soluble bases such as macrogol (polyethylene glycol), 1,3-butylene glycol, glycerol, gelatin, white sugar, sugar alcohol and the like; Emulsifiers such as glycerin fatty acid esters, polyoxyl stearate, and polyoxyethylene hardened castor oil; Acrylic acid ester, and sodium alginate; Jet agents such as liquefied petroleum gas and carbon dioxide; And preservatives such as p-hydroxybenzoic acid esters. The external preparation of the present invention can be prepared by using these methods according to a conventional method. In addition, a stabilizer, a flavoring agent, a coloring agent, a pH adjuster, a diluent, a surfactant, a preservative, an antioxidant and the like may be blended as needed. The use of the external preparation of the present invention can be applied to the top of the local window by a conventional method.

According to another aspect of the present invention, there is provided a method for producing a microorganism, comprising the steps of: treating a degreasing microalgae with an organic solvent to remove lipids and carbohydrates; A protein elution step of eluting the protein by treating a basic solution of the defatted microalgae in which the protein is concentrated; A protein precipitation step of treating the protein eluted in the protein elution step with an acidic solution to precipitate the protein; And a recovery step of removing the protein precipitated in the protein precipitation step and recovering a supernatant or a solid residue from the remaining defatted microalgae, wherein the crude polyphenol is extracted from the defatted microalgae.

In the extraction method of crude polyphenol from the degreasing microalgae, the organic solvent may be ethanol, methanol, acetone, ethyl acetate, ethylene glycol or benzene, and the acidic solution may be sulfuric acid, hydrochloric acid, nitric acid and acetic acid The basic solution may be calcium hydroxide (Ca (OH) 2), sodium hydroxide (NaOH) or ammonia.

According to another aspect of the present invention, There is provided a method for extracting crude polyphenols from degreasing microalgae, which comprises an extraction step of extracting the degreasing microalgae using water, an organic solvent or a mixture thereof or ultrasonic waves.

 In the method for extracting crude polyphenol from the degreasing microalgae, the ultrasonic wave may be applied at a frequency of 10 to 90 kHz under a temperature condition of 20 to 110 ° C.

The extraction of physiologically active substances from the degreasing microalgae accelerated the oxidation at high temperature, resulting in rapid degradation of the physico-chemical properties of the physiologically active substances, leading to the deterioration of the physiological activity. The problems such as protein mutation, high energy consumption and low extraction efficiency . Therefore, studies on techniques for preventing the degradation of physiologically active substances have been continuously tried. The present inventors have noted that, among various extraction techniques, ultrasonic extraction is a technique capable of minimizing loss of physiologically active substance under mild temperature and pressure. Ultrasonic extraction method has been recognized as an effective method for physiologically active substances from plant cell walls with high extraction efficiency and short extraction time compared to existing extraction methods. It is a technology that interest is increasing, and separation and transmission efficiency of physiologically active substances is improved due to the pupil phenomenon of ultrasonic waves Which is an effective process for extracting physiologically active substances from cell walls. Therefore, in order to minimize the destruction of physiologically active substances, ultrasonic extraction method (Ultrasonic-assisted extraction, UAE) was performed at a temperature of 100 ° C. or less using the defatted microalgae after extraction process to produce biodiesel from Tetraselmis , As a result of the internal pressure increase, destruction of the lignocellulosic structure, increase of the diffusion of the extract, and low temperature and short extraction time due to cavitation due to ultrasonic waves, it is more economical and whitening, antioxidant, anti-aging and skin Anti-aging and whitening cosmetic compositions derived from a degreasing microalga having excellent regenerating effect were developed.

Degreasing microalgae contain natural antioxidants including polyphenol compounds, and it contains a large amount of functional materials such as polysaccharides and antioxidants which are not eluted in the solvent used for lipid extraction. Therefore, it has high value as high value-added foods, cosmetics and pharmaceuticals have. Functional biologically active materials using natural biological materials with little harmful effects on humans and the environment have been developed and put into practical use. Therefore, it is necessary to search for new antioxidants from new plant resources, develop natural antioxidants such as material separation, And many plant extracts and herbal medicine extracts act on tyrosinase to inhibit melanin production. However, it has been found that there are many problems in using the drug at an effective concentration or higher in terms of stability and possibility of discoloration, But it has not yet shown an excellent effect. Development of a whitening functional cosmetic material utilizing the inhibitor of tyrosinase contained in the cell walls of the microfluidic microalgae enables the high value added of the microfluidic microbial cells to be discarded. The polyphenol and the physiologically active substance To develop antioxidant and anti-aging and functional cosmetic materials. Therefore, it is urgent to develop the field of the micro-algae extract for the limited field of research and commercialization.

Hereinafter, the present invention will be described in more detail by way of examples. It should be understood, however, that the invention is not limited to the disclosed embodiments, but may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, Is provided to fully inform the user.

Example 1: Preparation of defatted microalgae

The microalgae used in the present invention were obtained from a microalgae Tetraselmis KCTC 12236BP which had been degreased after cultivation in an indoor incubator at Inha University, and then dried in an oven at 60 ° C for 12 hours, Microalgae were used and kept frozen at 5 ℃ until used. The organic solvent used as the extraction solvent was a reagent having a purity of 95% or more of Sigma-Aldrich (St. Louis, Mo., USA) and the reagent used for the analysis was a reagent of first grade or higher from Sigma-Aldrich.

Example 2: Degreasing microalgae cleaning

For the protein extraction, a sample of defatted microalgae powder and 0.01 mol sulfuric acid solution were mixed at a ratio of 1:10 (w / w) and washed at 150 rpm for 2 hours in a shaking incubator at 30-50 ° C. to remove residual organic or acid Solution and other impurities were removed.

Example 3: Protein separation using a base / acid solution

The protein elution and precipitation using the base solution were carried out by mixing the defatted microalgae washed in Example 2 with NaOH at a ratio of 1:10 (w / w), and then, in a shaking incubator maintained at 30-100 ° C, Or by sonication at 20-100 kHz at 30-100 < 0 > C in an ultrasonic extractor. Thereafter, the mixture was centrifuged at 4,000 × g at 4 ° C. for 10 minutes, and the supernatant was recovered. The protein was precipitated by adding 5 N HCl to the isoelectric point and then centrifuged at 10,000 × g at 4 ° C. for 10 min. The precipitated protein was recovered from the recovered supernatant or precipitated solid residue Polyphenol was extracted and used.

Example 4: Protein separation using an organic solvent

The mixture was mixed at a ratio of 1:10 (w / w) using 20-100% organic solvent according to an embodiment of the present invention, and stirred at 150 rpm for 6 hours in a shaking incubator maintained at 30-100 ° C, And the non-protein portion was removed under the condition of 20-80 kHz at 30-100 ° C in an ultrasonic extractor. Thereafter, the mixture was centrifuged at 4,000 × g at 4 ° C. for 10 minutes to recover the supernatant, and the polyphenol was extracted from the recovered supernatant or the precipitated solid residue.

Example 5: Polyphenol extraction using ultrasonic

According to one embodiment of the present invention, prior to the extraction experiment using the defatted microalgae cryopreserved, the sample was dried in an oven at 60 ° C. until there was no change in the drying weight. The degreasing microalgae used in the present invention were dried after the removal of the lipids and the microalgae residues were used. For the extraction of polyphenol, a dry sample is added to a sealed glass tube at a ratio of 1: 5-1: 20, which is a basic extraction condition, and a solvent, and the mixture is stirred at 30-100 ° C for 1-60 minutes with water or an organic solvent Ultrasonic extraction was performed. The ultrasonic extraction was performed by adding ultrasonic waves (20-80 kHz, 200W) in a stand-bath mode using an ultrasonic machine (JAC Ultrasinic, Hwaseong, Korea) after adding distilled water to the degreasing microalgae. During extraction, the tube was placed in a stainless rack so that the bottom of the tube did not reach the bottom of the ultrasonic bath, and the extract and solvent in the container were placed so that they were completely immersed in the water. After extraction, 1 ml of the extract was transferred to a microtube, centrifuged, and the supernatant was recovered, and a diluent was prepared as needed to measure total polyphenol (TPC).

Example 6: Analysis of components of degreasing microalgae

Analysis of the components of the cell walls of the degreasing microalgae was carried out according to the NREL LAP (Laboratory Analytical Procedure) proposed by the US National Renewable Energy Laboratory (NREL). 0.3 g of the corresponding biomass solid sample was added to 3.0 ml of concentrated sulfuric acid (72% w / w), hydrolyzed at 30 ° C for 1 hour, diluted with 4.0 ml of 4.0% sulfuric acid in 84 ml of distilled water, Was subjected to secondary hydrolysis at 121 ° C for 1 hour. The hydrolyzed supernatant was quantitatively analyzed for each component using HPLC (Bio-Rad Aminex HPX-87P column) and Refractive index detector (Varian 356-LC, Varian, Inc., CA, USA). The cell wall components of the dried defatted microalgae Tetraselmis are shown in Table 1 below.

Dry weight ratio (%) cellulose Hemicellulose Lignin LEA 13.6 5.6 3.8 Rice straw 34.4 17.9 12.4 Corn stover 38.4 24.4 17.0 Switchgrass 31.0 21.2 17.6

Example 7: Total polyphenol (TPC) analysis

According to one embodiment of the present invention, the total polyphenol content was determined using the Folin-Denis method. Folin-Ciocalteu's phenol solution was added to the diluted extract and the polyphenol concentration was measured by the absorbance change based on the principle of reduction to blue color by the polyphenol compound. Each extract was diluted to 1 mg / mL, 0.125 mL of each diluted extract was added, and 0.5 mL of distilled water was added thereto. Then, 0.125 mL of Folin-Ciocalteu reagent was added and reacted for 5 minutes. Then, 1.25 mL of 7% sodium carbonate solution And the reaction was allowed to proceed at room temperature for 1 hour, and the absorbance was measured at 760 nm. The measured values were expressed as mg gallic acid equivalent (GAE) / g dry weight (DW) as compared to the standard curve using galic acid at each concentration. All samples were expressed using mean and standard deviation from three replicate experiments. All analyzes were repeated 3 times and the results were expressed as means and standard deviation (mean ± SD) and analyzed using SAS software (Statistical Analysis System, SAS Institute Inc., Cary, NC, USA) range test (p <0.05).

As a result, the total polyphenol concentration was the highest at microbial crushing size of 0.36 mm and 2.5% of the liquid fraction (FIGS. 1 and 2). When polyphenol production ability was evaluated according to ethanol concentration, which is a solvent used for ultrasonic extraction, polyphenol production was highest at an ethanol concentration of 60% (FIG. 3). In addition, when polyphenol production was compared according to the ultrasonic extraction temperature of the degreasing microalgae, the polyphenol production tended to increase proportionally with the increase of the ultrasonic extraction temperature. When the polyphenol production was carried out at 100 ° C, (Fig. 4). In addition, it was confirmed that the effect of ultrasonic extraction time on polyphenol production was significant, and it was confirmed that the increase of polyphenol production concentration after 20 minutes of extraction was not large, and 20 minutes of extraction time was set as optimal extraction time (FIG. 5) . The optimum conditions for the increase of polyphenol production from the degreasing microalgae were investigated. Optimization was carried out using temperature, time, and sulfuric acid concentration as the main variables using statistical optimization. As a result, The polyphenol of 8.04 g GAE / g DM was produced at an extraction temperature of 100 ° C, an extraction time of 46.8 minutes and a sulfuric acid concentration of 0.32 N (FIGS. 6 to 8). Using the optimal conditions obtained from the above experiment, the production of polyphenol, which is a direct indicator of antioxidant activity against ultrasonic extraction using weak acids, compared to hot water extraction of defatted microalgae was 4.80 mg GAE / g DW, whereas ultrasonic extraction 8.76 mg GAE / g DW extraction showed a 1.8-fold higher TPC content, indicating that the productivity of ultrasonic extraction was superior to that of conventional hot water extraction (FIG. 9).

Example 8: Evaluation of whitening efficacy

The synthesis of melanin for whitening efficacy evaluation is regulated by tyrosinase and its complex enzyme, and tyrosinase is a key enzyme of melanin biosynthesis as a rate - determining enzyme for melanin biosynthesis. Tyrosinase activity was determined by mixing 0.8 ml of 0.1 M potassium phosphate buffer (pH 6.8), 0.5 ml of 0.2 mg / ml L-tyrosine aqueous solution and 0.1 ml of 1000 unit / ml tyrosinase, Lt; 0 &gt; C for 20 minutes. The absorbance of the reaction solution was measured at 450 nm and the enzyme inhibitory activity of the sample was determined as a relative value to albumin as a positive control. To the reaction mixture, 0.2 mL of mushroom tyrosinase (110 U / mL) was added to 0.5 mL of 0.175 M sodium phosphate buffer (pH 6.8) and 0.2 mL of the substrate solution in which 10 mM L-DOPA had been dissolved and 0.1 mL of the sample solution. And the DOPA chrome produced in the reaction solution was measured at 475 nm. The tyrosinase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample.

As a result, the degreasing microalgae showed tyrosinase inhibitory activity in a concentration dependent manner and the tyrosinase inhibitory activity was higher at the same concentration than the simple microalgae extract. In particular, at a concentration of 50 ug / ml, almost the same activity as that of arbutin at the same concentration as that of the positive control was exhibited (FIG. 10).

In conclusion, the cosmetic composition prepared by the ultrasonic extraction method of the defatted microalgae, which is a byproduct remaining after the extraction process to produce the biodiesel from the microalgae Tetraselmis , has almost no cytotoxicity and has excellent antioxidative, whitening and anti Aging and skin regeneration, it can be usefully used for the development of cosmetics for skin whitening, external preparation for skin, pharmaceutical or functional foods, and production of products.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims.

Claims (8)

An anti-aging and whitening cosmetic composition derived from a defatted microalgae, which contains an extract of lipid-extracted microalgae as an active ingredient. The method according to claim 1,
Wherein the microalgae are tetracellis, spirulina, chlorella, or vegetable plaques, which are derived from defatted microalgae. A protein concentration step of treating degreasing microalgae with an organic solvent to remove lipids and carbohydrates;
A protein elution step of eluting the protein by treating a basic solution of the defatted microalgae in which the protein is concentrated;
A protein precipitation step of treating the protein eluted in the protein elution step with an acidic solution to precipitate the protein; And
And recovering the supernatant or the solid residue from the remaining defatted microalgae by removing the protein precipitated in the protein precipitation step and extracting the crude polyphenol from the defatted microalgae. The method of claim 3,
Wherein the organic solvent is ethanol, methanol, acetone, ethyl acetate, ethylene glycol or benzene; and extracting the crude polyphenol from the degreasing microalgae. The method of claim 3,
Wherein the acidic solution is sulfuric acid, hydrochloric acid, nitric acid, and acetic acid; and extracting the crude polyphenol from the degreasing microalgae. The method of claim 3,
Wherein the basic solution is calcium hydroxide (Ca (OH) ₂ ), sodium hydroxide (NaOH) or ammonia, and extracting the crude polyphenol from the degreasing microalgae. Preparing a degreasing microalgae;
A method for extracting crude polyphenols from degreasing microalgae, comprising extracting the degreasing microalgae using water, an organic solvent or a mixture thereof or ultrasonic waves. 8. The method of claim 7,
Wherein the ultrasonic wave is applied at a frequency of 10 to 90 kHz under a temperature condition of 20 to 110 캜, and extracting the crude polyphenol from the degreasing microalgae.

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