KR102463374B1 - A method of simultaneously extracting a non-polar component and a polar component from a natural product using two or more immiscible solvents, and a natural product extract manufactured by the method - Google Patents

Abstract

It relates to a method for simultaneously extracting a non-polar component and a polar component from a natural product using two or more immiscible solvents and a natural product extract prepared by the method, and the extraction method according to an aspect is harmful by extracting using two solvents that do not mix with each other. The extraction efficiency of lipids is high without the use of organic solvents, and phospholipids and neutral lipids can be recovered from each layer by evaporating the polar solvent after separating the two layers, so a complex post-process is required to separate neutral lipids and phospholipids. It can be used in food or cosmetics even in the form of total lipids including phospholipids and neutral lipids by evaporating the polar solvent without separation.

Description

A method of simultaneously extracting a non-polar component and a polar component from a natural product using two or more immiscible solvents and a natural product extract prepared by the method {A method of simultaneously extracting a non-polar component and a polar component from a natural product using two or more immiscible solvents, and a natural product extract manufactured by the method}

It relates to a method for simultaneously extracting a non-polar component and a polar component from a natural product using two or more immiscible solvents, and a natural product extract prepared by the method.

In order to extract an active ingredient from a natural product, a method of selecting a solvent in which the active ingredient dissolves well, impregnating it, and then filtering is generally used. Representative examples include extracting soybeans with hexane to produce soybean oil or extracting egg yolk with ethanol to obtain lecithin. Sometimes, like extracting polyphenols by mixing water and ethanol, it is sometimes extracted by mixing a solvent in which natural substances are easily soluble. After extraction, the solvent is removed and purified to obtain an active ingredient. In the case of soybeans, when extracted with hexane and the hexane is removed, gum is obtained along with soybean oil. The main component of gum is soybean lecithin, which is used to separate soybean oil and soybean lecithin through a degumming process that removes the gum.

Among the active ingredients of natural products, non-polar components are generally extracted by using solvents such as hexane, alcohol, or acetone or by pressing. In addition, compression extraction method or supercritical extraction method, which is a method that excludes harmful solvents, is also widely used (Korean Patent Application Laid-Open No. 2002-0064645).

To extract total lipids including phospholipids that are well soluble in ethanol or methanol, such as soybeans and egg yolk, and neutral lipids that are not soluble in ethanol but well soluble in chloroform, a Folch solution prepared with chloroform and methanol, two solvents that mix well with each other also use it Total lipids extracted with Folch solution or hexane are evaporated to obtain neutral lipids, phospholipids, and fat-soluble pigments. However, there is a problem in that environmental problems occur while using harmful organic solvents, and product preference is reduced due to concerns about residual solvents. In addition, the post-process of evaporating the solvent and separating the remaining total lipids into neutral lipids and phospholipids is complicated. Therefore, there is a need for a method capable of effectively extracting total lipids from natural products while excluding the use of harmful organic solvents.

One aspect is (A) extracting by adding a polar solvent and a non-polar solvent to the natural product and stirring; (B) filtering the extract to remove solids to obtain a filtrate; And (C) forming an immiscible non-polar solvent layer and a polar solvent layer by allowing the obtained filtrate to stand still; a method for simultaneously extracting a polar component and a non-polar component from a natural product using an immiscible solvent comprising a. will do

Another aspect is to provide a natural extract containing a non-polar component, a polar component, or a combination thereof prepared by the above method.

Another aspect is to provide a food composition comprising a natural product extract containing the non-polar component, the polar component, or a combination thereof.

Another aspect is to provide a cosmetic composition comprising a natural extract containing the non-polar component, the polar component, or a combination thereof.

One aspect is (A) extracting by adding a polar solvent and a non-polar solvent to the natural product and stirring; (B) filtering the extract to remove solids to obtain a filtrate; And (C) forming an immiscible non-polar solvent layer and a polar solvent layer by allowing the obtained filtrate to stand still; a method for simultaneously extracting a polar component and a non-polar component from a natural product using an immiscible solvent comprising a. do.

The natural product may be a vegetable or animal material. In one embodiment, the natural product may be any one selected from the group consisting of vegetable and animal natural products, including neutral lipids and phospholipids, and processed products thereof, for example, egg yolk powder, egg yolk powder, soybean , green-lipped mussels, krill, buttermilk powder, green bean coffee, cheonggukjang, rice buds, evening primrose seeds, ginseng, and red ginseng may be any one selected from the group consisting of. In one embodiment, the natural product may be a dried product obtained by natural drying, vacuum drying, drum drying, foam drying, hot air drying or freeze drying.

The term “polar solvent” may refer to a solvent whose molecular structure is itself a permanent electrical dipole. For example, the polar solvent may be a C1-C6 alcohol, water, acetone, ketone, (eg, methylethylketone), or a mixture thereof. In one embodiment, the C1-C6 alcohol is methanol, ethanol, n-propanol, isopropanol, n-butanol, sec-butanol, isobutanol, tert-butanol, n-pentanol, n-hexanol, 1, 2-hexanediol, 1,6-hexanediol, or a mixture thereof. The polar solvent may be, for example, a mixture of water and alcohol, that is, an aqueous alcohol solution. In one embodiment, the polar solvent has a purity of 20 to 99.9% (w/w), for example, 30 to 80% (w/w), 30 to 60% (w/w), 40 to 90% (w/w), 40-80% (w/w), 40-60% (w/w), 45-55% (w/w), 80-97% (w/w), 90-97% (w/w), 92-97% (w/w), 93-97% (w/w), 94-97% (w/w), 95-97% (w/w), 90-96% (w/w), 92 to 96% (w/w), 93 to 96% (w/w), 94 to 96% (w/w), 95 to 96% (w/w). Depending on the type of natural product, if the purity of ethanol is less than 30% (w/w) or more than 60% (w/w), the yield of antioxidants may be lowered, and the purity of ethanol is less than 92% (w/w) or When it exceeds 97% (w/w), the yield of phospholipids may be lowered, and when the purity of ethanol is less than 40% (w/w) or more than 90% (w/w), the yield of polyphenol may be lowered.

The term “non-polar solvent” may refer to a solvent in which the solvent molecules do not have permanent dipoles and, as a result, have no power to cause intermolecular associations with polar species. For example, the non-polar solvent may be vegetable, mineral, and animal oils or butter, for example, medium chain triglycerides (MCT), egg yolk oil, evening primrose oil, soybean oil, mineral oil, butter, squalane, squalene, shea butter oil, olive oil, coconut oil, palm oil, grapeseed oil, sunflower oil, jojoba oil, beeswax, sweet almond oil, castor oil, algan oil, avocado oil, cocoa butter, or mixtures thereof.

The polar solvent and the non-polar solvent are immiscible solvents, and each solvent may form a layer when mixed. Therefore, when a mixed solvent of a polar solvent and a non-polar solvent is used as an extraction solvent for a natural product, an active ingredient that is easily soluble in each solvent may be simultaneously extracted into each solvent layer. For example, phospholipids, which are polar components that are easily soluble in polar solvents, are dissolved in the polar solvent layer, and neutral lipids, which are non-polar components that are easily dissolved in non-polar solvents, are dissolved in the non-polar solvent layer to form an immiscible layered structure. can be simultaneously extracted, and the separation of both may be easy.

The agitation is rotational stirring, vertical reciprocating stirring (RS-1, Jeotech), vertical rotational stirring (CRT-350, Jeotech), orbital stirring (CPS-350, Jeotech), or shaking stirring, vibration stirring, rocking stirring (CWS-350, Jeotech) 1 to 1500 rpm (revolution / min), for example, 20 to 1000 rpm, 50 to 800 rpm, 100 to 600 rpm, may be stirred at a speed of 200 to 400 rpm. If the stirring speed is less than 1 rpm (revolution/min), the stirring effect may be reduced.

The extraction may be extracted by a conventional method in the art, such as room temperature extraction, cooling extraction, warming extraction, pressure extraction, microwave extraction, ultrasonic extraction, hot water extraction, reflux cooling extraction, subcritical extraction, or supercritical extraction. have. The extraction is -20 °C to 90 °C, for example, -20 °C to 70 °C, 2 °C to 30 °C, 20 °C to 30 °C, 20 °C to 70 °C, 20 °C to 60 °C, 20 °C to 50 °C , 20 ℃ to 40 ℃, 37 ℃ to 60 ℃, it may be carried out at 37 ℃ to 50 ℃. When the extraction temperature is less than 2 ℃, the lipid extraction efficiency may be reduced, and if it is more than 90 ℃, the non-polar solvent may be evaporated.

The extraction time may be appropriately selected according to the selected temperature and extraction method. For example, the extraction time may be 10 minutes to 60 days, 10 minutes to 14 days, 10 minutes to 7 days, 10 minutes to 48 hours, 10 minutes to 24 hours, 10 minutes to 10 hours, 10 minutes to 2 hours, 20 minutes to 2 hours, 30 minutes to 8 hours, 30 minutes to 4 hours, 12 hours to 48 hours, 12 hours to 24 hours, or 24 hours to 48 hours. If the extraction time is less than 10 minutes, the lipid extraction efficiency may be reduced, and if the extraction time is more than 60 days, it may lead to oxidation of the lipids.

The extraction may be one or more times, for example, extracted 1 to 5 times, 1 to 4 times, 1 to 3 times, 2 to 5 times, or 2 to 4 times, and each extraction is performed in the same way, It can be done in other ways.

In one embodiment, the step (A) is based on 100 parts by weight of the natural product, 100 to 800 parts by weight of a polar solvent, for example, 100 to 500 parts by weight, 100 to 400 parts by weight, 100 to 200 parts by weight, 200 parts by weight. to 500 parts by weight, 200 to 400 parts by weight, and 10 to 1600 parts by weight of a non-polar solvent, for example, 10 to 800 parts by weight, 100 to 400 parts by weight, 100 to 200 parts by weight, 10 to 100 parts by weight, 15 to 50 parts by weight, 15 to 25 parts by weight may be added. Specifically, based on 100 parts by weight of the natural product, 300 to 500 parts by weight of a polar solvent, and 150 to 250 parts by weight of a non-polar solvent may be added.

In one embodiment, the step (B) may be to filter the extract through a mesh of 10-200 mesh. Filtration methods may vary depending on the type of natural products such as dashi bags, sieves, filter papers, and sieves. The filtration may be one or more times, for example, filtered 1 to 5 times, 1 to 4 times, 1 to 3 times, 2 to 5 times, or 2 to 4 times, and each filtration is performed in the same way, It can be done in other ways. In one embodiment, the method according to claim 1, after the step (C), separating the formed non-polar solvent layer and the polar solvent layer to obtain a non-polar component-containing extract; and evaporating the polar solvent from the separated polar solvent layer to obtain an extract containing a polar component; may further include. The method may further include; purifying the obtained non-polar component-containing extract to recover the non-polar active ingredient. In addition, the method may further include; purifying the obtained extract containing the polar component to recover the polar active ingredient. In another embodiment, after step (C), evaporating the polar solvent without separating the non-polar solvent layer and the polar solvent layer to obtain an extract containing a non-polar component and a polar component; may further include .

In one embodiment, the non-polar component may include at least one selected from the group consisting of neutral lipids, carotenoids, and lutein.

In one embodiment, the polar component may include at least one selected from the group consisting of phospholipids, polyphenols, and ginsenosides.

The extraction method according to an aspect can be used to separate the two solvent layers to recover phospholipids and neutral lipids from each solvent layer, respectively, and can also be used in the form of total lipids including phospholipids and neutral lipids without separating the two solvent layers. can

The stationary is for layer separation, and the stationary time is, for example, 10 minutes to 7 days, 10 minutes to 18 hours, 10 minutes to 12 hours, 10 minutes to 6 hours, 10 minutes to 3 hours, 10 minutes to 2 hours, 30 minutes to 12 hours, 30 minutes to 6 hours, 30 minutes to 3 hours, 30 minutes to 2 hours, 3 hours to 12 hours, 6 hours to 12 hours, or 6 hours to 9 hours.

The polar solvent may be removed by natural drying, vacuum drying, drum drying, foam drying, hot air drying, or freeze drying.

Another aspect is to provide a natural extract containing a non-polar component, a polar component, or a combination thereof prepared by the above method. In the natural extract containing the non-polar component, the polar component, or a combination thereof, the preparation method, the non-polar component, the polar component, the natural product, extraction, etc. are as described above. The natural extract containing a non-polar component, a polar component, or a combination thereof according to an aspect may be an egg yolk powder extract containing neutral lipids and phospholipids prepared by the above method, that is, an extract containing total lipids. The total lipid-containing extract prepared by the above method may contain a larger amount of total lipids than when a single solvent is used or a Folch solvent that is well mixed with each other is used. The total lipid-containing extract may further include other active ingredients soluble in non-polar solvents and polar solvents in addition to neutral lipids and phospholipids.

Another aspect is to provide a natural extract containing a non-polar component prepared by the above method. The natural extract containing a non-polar component according to an aspect may be a neutral lipid-containing egg yolk extract prepared by the above method, that is, an extract containing egg yolk oil. The neutral lipid-containing extract prepared by the above method may contain a larger amount of neutral lipids than when a single solvent is used or a Folch solvent that is well mixed with each other is used. The neutral lipid-containing extract may further include other active ingredients soluble in a non-polar solvent in addition to the neutral lipid.

Another aspect is to provide a natural extract containing the polar component prepared by the above method. The natural product extract containing the polar component according to an aspect may be a phospholipid-containing egg yolk extract prepared by the above method, that is, an extract containing egg yolk lecithin. The phospholipid-containing extract prepared by the above method may contain a larger amount of phospholipids than when a single solvent is used or a Folch solvent that is well mixed with each other is used. The phospholipid-containing extract may further include other active ingredients soluble in a polar solvent in addition to phospholipids.

Another aspect is to provide a food composition comprising a natural product extract containing the non-polar component, the polar component, or a combination thereof. In the food composition, the manufacturing method, non-polar component, polar component, natural product, extraction, etc. are the same as described above.

The "food composition" may be used alone or in combination with other foods or food ingredients, and may be appropriately used according to a conventional method, using a natural extract containing a non-polar component, a polar component, or a combination thereof prepared by the above method. . The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prophylactic, health or therapeutic treatment). In general, when preparing food or beverage, the food composition may be added in an amount of 15 parts by weight or less based on the raw material. There is no particular limitation on the type of the food. Among the types of food, the beverage composition may contain various flavoring agents or natural carbohydrates as an additional component like a conventional beverage. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatine and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The food composition may also contain nutrients, vitamins, electrolytes, flavoring agents, antioxidants, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonic acid Carbonating agents used in beverages, or a combination thereof. The food composition may also contain natural fruit juice, fruit juice beverage, fruit flesh for the preparation of vegetable beverage, or a combination thereof.

Another aspect is to provide a cosmetic composition comprising a natural extract containing the non-polar component, the polar component, or a combination thereof. In the cosmetic composition, the preparation method, non-polar component, polar component, natural product, extraction, etc. are the same as described above.

The "cosmetic composition" can be prepared in various forms, and in one embodiment, the composition is an emulsion, lotion, cream (oil-in-water type, water-in-oil type, multi-phase), solution, suspension (anhydrous and water-based), It may be in any one formulation selected from the group consisting of emulsions, anhydrous products (oil and glycol-based), pastes, gels, masks, packs, powders, soaps, powders, and sprays.

The cosmetic composition may further include an acceptable carrier in the cosmetic formulation. The "acceptable carrier for cosmetic preparations" refers to compounds or compositions that are already known and used that can be included in cosmetic preparations, or compounds or compositions to be developed in the future that do not have toxicity, instability, or irritation beyond what the human body can adapt when in contact with the skin. say that The carrier may be included in the cosmetic composition of the present specification in an amount of about 1% to about 99.99% by weight, preferably about 90% by weight to about 99.99% by weight of the total weight of the composition. Examples of the carrier include alcohol, oil, surfactant, fatty acid, silicone oil, humectant, humectant, viscosity modifier, emulsifier, stabilizer, sunscreen, color developer, fragrance, and the like.

The extraction method according to an aspect has high extraction efficiency of lipids without using harmful organic solvents by extracting using two immiscible solvents, and phospholipids and neutral lipids from each layer by evaporating the polar solvent after separating the two layers Since they can be recovered individually, a complicated post-process for separating neutral lipids and phospholipids is not required, and by evaporating the polar solvent without separation, it can be used in food or cosmetics in the form of total lipids including phospholipids and neutral lipids.

1 is a ternary plot graph of the weight ratio of egg yolk powder, ethanol, and MCT mixed to prepare an ethanol/medium chain neutral fat mixed solvent extract of egg yolk powder of Example 1-2.
2 is a photograph confirming the properties of an ethanol/medium chain triglyceride mixed solvent extract of egg yolk powder prepared by varying the weight ratios of egg yolk powder, ethanol, and medium chain triglycerides in Example 1-2.
3 is a photograph confirming the properties of a mixed extract of ethanol/medium chain triglycerides of egg yolk powder of Example 1-1.
4 is a photograph confirming the properties of the egg yolk powder extract obtained in Example 1-1 and Comparative Examples 1-1 to 1-4.
5 is an egg yolk extract extracted using the solvent of Example 1-2 prepared in various weight ratios of Example 1-1 and the ethanol/MCT mixed solvent and the solvent of Comparative Examples 1-1 to 1-4. This is the result of quantifying the fatty acid content of the egg yolk powder extract based on 100 g of the dried raw material.
6 is an egg yolk extract extracted using the solvent of Example 1-2 prepared in various weight ratios of Example 1-1 and the ethanol/MCT mixed solvent and the solvent of Comparative Examples 1-1 to 1-4. This is the result of quantifying the total phospholipid content of the egg yolk powder extract by weight ratio of acetone insolubles based on 100 g of the dry raw material.
7 is a result of quantifying the weight ratio of acetone insolubles based on 100 g of dry raw material for the total phospholipid content of egg yolk powder extracts obtained by varying the purity of ethanol of Example 1-1 to 70, 90, 95, and 99.9% (w/w) .
8 is a photograph confirming the properties of the extracts of Examples 1-3 and Comparative Examples 1-5.
9 is a photograph in which the butter extract prepared in Examples 1-2 and Comparative Example 1-5 in Experimental Example 4.2 was mixed with water in a weight ratio of 1:9, mixed with a hand mixer, and then phase separation was observed with the naked eye. .
10 shows the total phospholipid content of the egg yolk powder extract extracted using the solvents of Examples 1-1 and 1-3 and the egg yolk powder extract extracted using the solvents of Comparative Examples 1-4 and 1-5 based on 100 g of acetone-insoluble This is the result of quantifying the weight ratio of water.
11 is a graph analyzing the lutein content of the egg yolk powder extract extracted using the solvents of Examples 1-1 and 1-3 and the egg yolk powder extract extracted using the solvents of Comparative Examples 1-4 and 1-5.
12 is a graph analyzing the lutein content of an egg yolk powder extract extracted using the solvent of Example 1-1 and an egg yolk powder extract extracted using the solvent of Comparative Examples 1-1 to 1-4.
13 is a photograph confirming the properties of the mixed solvent extract of ethanol/medium chain neutral fat of various natural products of Example 2-1.
14 is a photograph confirming the properties of the ethanol solvent extract of various natural products of Comparative Example 2-2.
15 shows the fatty acid content of extracts of various natural products extracted using the ethanol/MCT mixed solvent of Example 2-1 and the extracts of various natural products extracted using the solvents of Comparative Examples 2-1 and 2-2 based on 100 g of dry raw material is the result of quantification.
Figure 16 shows the total phospholipid content of various natural products extracted using the ethanol/MCT mixed solvent of Example 2-1 and the ethanol solvent of Comparative Example 2-2, the total phospholipid content of acetone insolubles based on 100 g of dry raw material. This is the result of quantifying the weight ratio.
17 is a diagram showing a mixture of yolk powder, krill, buttermilk, egg yolk powder, and green-lipped mussel powder extracts extracted using the ethanol single solvent of Comparative Example 2-2 or the ethanol/MCT mixed solvent of Example 2-1 with water 1: After mixing in a weight ratio of 9, it is a photograph of visual observation of phase separation after mixing with a hand mixer.
18 is a diagram showing a mixture of green-lipped mussel powder extract with water in a weight ratio of 1:9, and then mixed with water and a hand mixer. It is the result of observing the phase separation under a microscope.
19 shows the total polyphenol content of the evening primrose seed extract extracted using the ethanol/water/MCT mixed solvent of Example 2-2 and the evening primrose seed extract extracted using the ethanol/water solvent of Comparative Example 2-4 as a dry raw material It is the result of quantification based on 100g.
20 is an extract of evening primrose seeds extracted using the ethanol/water/MCT mixed solvent of Example 2-2 (month 50:50 ethanol/water layer, moon 50:50 total extract), ethanol/water of Comparative Example 2-2 Evening primrose seed oil extracted using solvent (1:4:0, 50%), tocopherol (JB, Mixed Tocopherol T-70), and evening primrose oil extracted by cold pressing from evening primrose seeds (Saero Hands, Evening Primrose Oil Refined) ) as a result of quantifying the 50% inhibitory concentration (IC ₅₀ ) in ppm, FIG. 20a is a general graph, and FIG. 20b is a log graph.
21 shows the ginsenoside content of the ethanol/water layer of the extract of red ginseng extracted using the ethanol/water/MCT mixed solvent of Example 2-2 and the extract of red ginseng extracted using the ethanol/water solvent of Comparative Example 2-4. is the result of quantification based on 1 g of dry raw material.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes of the present invention, and the scope of the present invention is not limited to these examples.

Example 1-1: Preparation of egg yolk powder extract using ethanol/medium chain triglycerides (MCT) mixed solvent

Egg yolk powder (Poonglim Food, egg yolk powder), ethanol (Daehan Alcoholic Life, 93.7% (w/w) alcoholic ethanol), and MCT (Lotte Foods, medium chain neutral fat) were mixed in a 1:4:2 weight ratio (ethanol weight in the extraction solvent 66.7 %) and shaken at 80 rpm for 1 hour (Roto-bot R4040, Benchmark Scientific, Sayreville, NJ, USA) for mixed extraction. After standing for 10 minutes, the egg yolk component was removed using a vacuum filter, and the filtered solution was evaporated by ethanol using a vacuum vacuum concentrator (Rotavapor N-1000, EYELA, Bohemia, NY, USA) to obtain an extract.

Example 1-2: Preparation of egg yolk powder extract using ethanol/medium chain triglycerides (MCT) mixed solvent

Egg yolk powder (Poonglim Food, egg yolk powder), ethanol (Daehan Alcoholic Life, 93.7% (w/w) alcohol ethanol), and MCT (Lotte Foods, medium chain triglyceride) were added in a 1:4:2 weight ratio instead of as [Table 1]. An ethanol/medium chain neutral fat mixed solvent extract of egg yolk powder was prepared in the same manner as in Example 1-1 except for changing the composition ratio.

Sample
ID parts by weight weight % yolk powder ethanol MCT yolk powder ethanol MCT 1:2:2 100 200 200 20 40 40 1:2:4 100 200 400 14 29 57 1:2:6 100 200 600 11 22 67 1:2:8 100 200 800 9 18 73 1:2:10 100 200 1000 8 15 77 1:2:12 100 200 1200 7 13 80 1:2:14 100 200 1400 6 12 82 1:2:16 100 200 1600 5 11 84 1:4:0.2 100 400 20 19 77 4 1:4:0.5 100 400 50 18 73 9 1:4:1 100 400 100 17 67 17 1:4:2 100 400 200 14 57 29 1:4:4 100 400 400 11 44 44 1:4:6 100 400 600 9 36 55 1:4:8 100 400 800 8 31 62 1:4:10 100 400 1000 7 27 67 1:4:12 100 400 1200 6 24 71 1:4:14 100 400 1400 5 21 74 1:4:16 100 400 1600 5 19 76 1:6:2 100 600 200 11 67 22 1:6:4 100 600 400 9 55 36 1:6:6 100 600 600 8 46 46 1:6:8 100 600 800 7 40 53 1:6:10 100 600 1000 6 35 59 1:6:12 100 600 1200 5 32 63 1:6:14 100 600 1400 5 29 67 1:6:16 100 600 1600 4 26 70 1:8:2 100 800 200 9 73 18 1:8:4 100 800 400 8 62 31 1:8:6 100 800 600 7 53 40 1:8:8 100 800 800 6 47 47 1:8:10 100 800 1000 5 42 53 1:8:12 100 800 1200 5 38 57 1:8:14 100 800 1400 4 35 61 1:8:16 100 800 1600 4 32 64

1 is a ternary plot graph of the weight ratio of egg yolk powder, ethanol, and MCT mixed to prepare an ethanol/medium chain neutral fat mixed solvent extract of egg yolk powder of Example 1-2. (The red dot is the ratio used for fatty acid analysis in Experimental Example 2.1, and the blue circle is the sample used in Experimental Example 2.2. Total phospholipid analysis.)

Example 1-3: Preparation of egg yolk powder extract using ethanol/oil mixed solvent

Use egg yolk oil, evening primrose oil, soybean oil, mineral oil, butter, squalane, shea butter oil, olive oil, coconut oil, palm oil, grapeseed oil, sunflower oil, or jojoba oil instead of MCT, butter, shea butter, palm oil, coconut oil An ethanol/oil mixed solvent extract of egg yolk powder was prepared in the same manner as in Example 1-1, except that the solid oil was extracted at 50°C.

Comparative Example 1-1: Preparation of egg yolk powder extract using methanol/chloroform mixed solvent (Folch solution)

Egg yolk powder (Poonglim Food, egg yolk powder) was mixed with Folch solution, a mixed solvent of 2:3 volume ratio of methanol and chloroform in a 1:4 weight ratio instead of ethanol/MCT mixed solvent, except that the Folch solution was evaporated instead of ethanol prepared an egg yolk powder extract in the same manner as in Example 1-1. The Folch solution is a mixture of chloroform (Samjeon Pure Yak, 99.5%, CAS No. 67-66-3) and methanol (Samjeon Pure Yak, 99.9%, CAS No. 67-56-1), which are miscible solvents, in a volume ratio of 3:2. It was prepared by mixing.

Comparative Example 1-2: Preparation of egg yolk powder extract using ethanol solvent

An egg yolk powder extract was prepared in the same manner as in Comparative Example 1-1, except that an ethanol (Daehan Alcoholic Life, 93.7% (w/w) alcoholic ethanol) solvent was used instead of the Folch solution.

Comparative Example 1-3: Preparation of egg yolk powder extract using hexane solvent

An egg yolk powder extract was prepared in the same manner as in Comparative Example 1-1, except that a hexane (Samjeon Soonyak, 95.0%, CAS No 110-54-3) solvent was used instead of the Folch solution.

Comparative Example 1-4: Preparation of egg yolk powder extract using MCT solvent

Comparative Example 1-1 except that an MCT (Lotte Foods, medium-chain triglyceride) solvent was used instead of the Folch solution, and only the removal of the egg yolk component was performed using a vacuum filter without evaporating the solvent using a vacuum concentrator. An egg yolk powder extract (MCT/yolk oil solution) was prepared in the same manner as described above.

Comparative Example 1-5: Preparation of egg yolk powder extract using oil solvent

Use egg yolk oil, evening primrose oil, soybean oil, mineral oil, butter, squalane, shea butter oil, olive oil, coconut oil, palm oil, grapeseed oil, sunflower oil, or jojoba oil instead of Folch solution, and use a vacuum concentrator to remove the solvent. An oil solvent extract of egg yolk powder was prepared in the same manner as in Comparative Example 1-1, except that only the removal of the egg yolk component was performed using a reduced pressure filter without an evaporation process.

Example 2-1: Preparation of ethanol/medium chain triglyceride mixed solvent extracts of various natural products

In the same manner as in Example 1-1, except that various natural products (abed egg yolk powder, soybean, green lipped mussel, krill, buttermilk powder, green bean coffee, cheonggukjang, rice bran, evening primrose seed, ginseng, and red ginseng) were used instead of egg yolk powder. Ethanol/medium chain neutral fat mixed solvent extracts of each natural product were prepared.

Example 2-2: Preparation of mixed solvent extracts of ethanol/water/medium chain neutral fat of various natural products

Instead of using evening primrose seeds or red ginseng as a natural product, and mixing natural products, ethanol (Korea Alcoholic Life, 93.7% (w/w) alcohol ethanol), and MCT (Lotte Foods, medium chain triglycerides) in a weight ratio of 1:4:2 As shown in [Table 2], natural products, ethanol (0 ~ 100% (w / w)), and MCT are mixed in a weight ratio of 1:4:1 (that is, the composition ratio of natural products, ethanol, and MCT and ethanol A mixed solvent extract of ethanol/water/medium chain neutral fat of each natural product was prepared in the same manner as in Example 2-1, except that the purity was changed.

Sample
ID parts by weight weight % natural product ethanol water MCT natural product ethanol water MCT month0:100 100 0 400 100 17 0 67 17 Moon 10:90 100 40 360 100 17 7 60 17 Moon 20:80 100 80 320 100 17 13 53 17 Moon 30:70 100 120 280 100 17 20 47 17 Moon 40:60 100 160 240 100 17 27 40 17 Moon 50:50 100 200 200 100 17 33 33 17 Moon 60:40 100 240 160 100 17 40 27 17 Moon 70:30 100 280 120 100 17 47 20 17 Moon 80:20 100 320 80 100 17 53 13 17 Moon90:10 100 360 40 100 17 60 7 17 Moon 93:7 100 372 28 100 17 62 5 17 Moon 95:5 100 380 20 100 17 63 3 17 moon100:0 100 400 0 100 17 67 0 17 Red0:100 100 0 400 100 17 0 67 17 Hong 10:90 100 40 360 100 17 7 60 17 Hong 20:80 100 80 320 100 17 13 53 17 Hong 30:70 100 120 280 100 17 20 47 17 Red 40:60 100 160 240 100 17 27 40 17 Red 50:50 100 200 200 100 17 33 33 17 Red 60:40 100 240 160 100 17 40 27 17 Red 70:30 100 280 120 100 17 47 20 17 Red 80:20 100 320 80 100 17 53 13 17 Red 90:10 100 360 40 100 17 60 7 17 Hong 93:7 100 372 28 100 17 62 5 17 Hong 95:5 100 380 20 100 17 63 3 17 Red100:0 100 400 0 100 17 67 0 17

Comparative Example 2-1: Preparation of solvent extracts in methanol/chloroform mixed solvent (Folch solution) of various natural substances

Except that the various natural products of Example 2-1 were mixed with Folch solution, which is a mixed solvent of 2:3 volume ratio of methanol and chloroform, in a weight ratio of 1:6 instead of the mixed solvent of ethanol/medium chain neutral fat, Example 2-1 and Folch solution extracts of each natural product were prepared in the same manner.

Comparative Example 2-2: Preparation of ethanol solvent extracts of various natural products

An ethanol solvent extract of each natural product was prepared in the same manner as in Comparative Example 2-1, except that an ethanol (Daehan Alcoholic Life, 93.7% (w/w) alcoholic ethanol) solvent was mixed in a weight ratio of 1:4 instead of the Folch solution. did.

Comparative Example 2-3: Preparation of MCT solvent extracts of various natural products

Instead of Folch solution, MCT (Lotte Foods, medium chain triglyceride) solvent was used in a weight ratio of 1:4, and only the removal of egg yolk was performed using a vacuum filter without evaporating the solvent using a vacuum concentrator. prepared MCT solvent extracts of each natural product in the same manner as in Comparative Example 2-1.

Comparative Example 2-4: Preparation of ethanol/water solvent extracts of various natural products

Instead of using evening primrose seeds or red ginseng as a natural product, and mixing a natural product and ethanol (Korea Alcoholic Life, 93.7% (w/w) alcoholic ethanol) in a weight ratio of 1:4, as in [Table 2] above, natural product and ethanol (0 ~ 100% (w/w)) of each natural product in the same manner as in Comparative Example 2-2, except that the mixture was used in a 1:4 weight ratio (that is, the purity of ethanol was used differently). An extract was prepared.

Experimental Example 1: Analysis of yield and properties of egg yolk powder extract according to extraction solvent

The following [Table 3] shows the results of measuring the weight of the egg yolk powder extract obtained in Example 1-1 and Comparative Examples 1-1 to 1-4 and confirming the yield according to the extraction solvent. The extracts of Example 1-1 and Comparative Example 1-4 were excluded from the weight of the extract after determining the MCT content through fatty acid analysis. Yield (%) was calculated as (weight of extract excluding MCT (g)/weight of sample (g))*100.

menstruum Sample weight (g) Extract weight (g) Yield (%) Ethanol/MCT (Example 1-1) 20 44.4 58.4 Folch (Comparative Example 1-1) 30 14.4 47.9 Ethanol (Comparative Example 1-2) 30 5.7 19.1 Hexane (Comparative Example 1-3) 30 11.0 36.8 MCT (Comparative Example 1-4) 30 105.1 39.6

As a result, the weight of the ethanol/medium chain neutral fat mixed solvent extract through 20 g of egg yolk powder was 44.4 g, and the yield of the extract excluding MCT was 58.4%, which was higher than Comparative Examples 1-1 to 1-4. Therefore, it can be seen that when extraction is performed using a mixed solvent of ethanol/medium chain neutral fat, which is an immiscible solvent, an extract with a yield of 1.2 to 3 times higher than when using a single solvent or a miscible solvent (Folch) can be obtained. It is possible to extract a large amount of active ingredients in egg yolk powder using an eco-friendly solvent that can be used for food and cosmetics without using the existing harmful organic solvents such as Folch or hexane, so the extraction efficiency can be improved in an eco-friendly way. This means that it can be significantly increased.

2 is a photograph confirming the properties of an ethanol/medium chain triglyceride mixed solvent extract of egg yolk powder prepared by varying the weight ratios of egg yolk powder, ethanol, and medium chain triglycerides in Example 1-2. 3 is a photograph confirming the properties of a mixed extract of ethanol/medium chain triglycerides of egg yolk powder of Example 1-1. As a result, as shown in FIG. 2, the color of the extract and the amount of gel were changed according to the weight ratio of the mixed solvent. Specifically, in the case of 1:4:0.2 with a low ratio of medium chain triglycerides, a yellow gel with strong viscosity was obtained as a single phase, and it was confirmed that liquid egg yolk lecithin, an extract containing egg yolk lecithin as the main component and egg yolk oil as a minor component, was prepared. did. In this case, since egg yolk oil is mixed, the viscosity of egg yolk lecithin is reduced, so post-processing is easy and it is convenient to apply to the process. On the other hand, in the case of 1:4:8 with a high ratio of medium chain triglycerides, it was confirmed that yellow and transparent oil was obtained as a single phase, and egg yolk oil containing lecithin, an extract containing egg yolk lecithin as a minor component and egg yolk oil as a main component, was prepared. did. In this case, since it contains lecithin, a natural emulsifier, when applied to food, it has the advantage of easy processing. In addition, in the case of 1:4:0.5, 1:4:1, and 1:4:2 (Example 1-1, FIG. 3), the properties of the extract are all yellow and transparent liquid in the form of sunken gel, and the gel-like lecithin As it was confirmed that the layer separation of the liquid egg yolk oil and the liquid egg yolk oil occurred, it was confirmed that the two layers could be separated and used or shaken and used in the post-process when prepared at the above ratio.

4 is a photograph confirming the properties of the egg yolk powder extract obtained in Example 1-1 and Comparative Examples 1-1 to 1-4. As a result, as shown in FIG. 4, in Comparative Example 1-1 extracted using Folch solution and Comparative Example 1-3 extracted using a hexane solvent, the oil exhibited the same properties, and Comparative Example extracted using an ethanol solvent In the case of 1-2, a gel-like appearance was exhibited, and in Comparative Example 1-4 extracted using the MCT solvent, the appearance of a yellow and transparent oil was exhibited. The ethanol/medium chain neutral fat mixed solvent extract of the egg yolk powder obtained in Example 1-1 had a gel-like state in a yellow and transparent liquid, and the ethanol extract of Comparative Example 1-2 and the MCT extract of Comparative Example 1-4 were mixed. It was confirmed that the shape was similar. This means that when extraction is performed using a mixed solvent of ethanol/medium-chain neutral fat, which is an immiscible solvent, active ingredients dissolved in each solvent layer can be simultaneously extracted.

Experimental Example 2: Analysis of active ingredient content of egg yolk powder extract according to extraction solvent

2.1. Fatty Acid Analysis

Fatty acid analysis was performed according to the 2.1.5.4 fatty acid analysis method of the Food Standards Code, except that heptane was used instead of isooctane. Agilent, GC-FID, HP 5890 was used as an analysis instrument.

Specifically, Example 1-2 prepared in various weight ratios of Example 1-1 and an ethanol/MCT mixed solvent, and an egg yolk extract using the solvents of Comparative Examples 1-1 to 1-4 were compared and analyzed for fatty acid content. . In the fatty acid analysis result, fatty acids except for C8:0 and C10:0 corresponding to MCT were quantified based on 100 g of dry raw material. 5 is an egg yolk extract extracted using the solvent of Example 1-2 prepared in various weight ratios of Example 1-1 and the ethanol/MCT mixed solvent and the solvent of Comparative Examples 1-1 to 1-4. This is the result of quantifying the fatty acid content of the egg yolk powder extract based on 100 g of the dried raw material.

As a result, as shown in FIG. 5, Examples 1-1 and 1-2 extracted using an ethanol/MCT mixed solvent had a section in which the fatty acid value was extracted higher than Comparative Examples 1-1 to 1-4. appeared to be This means that active ingredients such as egg yolk oil and egg yolk lecithin can be more efficiently extracted from egg yolk powder of the same weight during mixed extraction. In addition, it is an eco-friendly method by extracting non-polar and amphoteric substances without the use of harmful solvents such as Folch or hexane. Triglyceride, which is a neutral lipid, is often used in post-processing in food and cosmetic manufacturing, and it can be industrially usefully used to manufacture semi-finished products obtained by extracting active ingredients from natural products such as egg yolk using triglycerides and ethanol.

2.2. Total phospholipid analysis

Total phospholipids were analyzed by a physicochemical test method that measures the weight of acetone insolubles according to Ⅲ.3.5.2 Phospholipids (as acetone insolubles) of the Health Functional Foods Code.

Specifically, comparative analysis of the total phospholipid content with the egg yolk extract using the solvents of Comparative Examples 1-1 to 1-4 in Example 1-2 prepared in various weight ratios of Example 1-1 and ethanol/MCT mixed solvent did. The total phospholipid content was quantified by weight ratio of acetone insolubles based on 100 g of dry raw material. 6 is an egg yolk extract extracted using the solvent of Example 1-2 prepared in various weight ratios of Example 1-1 and the ethanol/MCT mixed solvent and the solvent of Comparative Examples 1-1 to 1-4. This is the result of quantifying the total phospholipid content of the egg yolk powder extract by weight ratio of acetone insolubles based on 100 g of the dry raw material.

As a result, as shown in FIG. 6, Examples 1-1 and 1-2 extracted using an ethanol/MCT mixed solvent had a higher total phospholipid value than Comparative Examples 1-1 to 1-4. appeared to be

Summarizing the above results, when extracting using a mixed solvent of ethanol/MCT, neutral lipids and phospholipids can be extracted at a higher yield than when using a single solvent, and a mixed solution of methanol/chloroform that mixes well with each other Compared to the case of using the Folch solution, it means that the extraction rate is similar without the use of harmful solvents. Lecithin is often used in food processing, but when extracted in this way, it can be developed as a food additive with high extraction efficiency and no residual solvent.

Experimental Example 3: Total phospholipid analysis of ethanol/medium chain triglyceride mixed solvent extract of egg yolk powder according to ethanol purity

Total phospholipids in the same manner as in Experimental Example 2.2, except that the ethanol/medium chain triglyceride mixed solvent in which the purity of the ethanol of Example 1-1 was variously changed to 70, 90, 95, and 99.9% (w/w) was used. Analysis was performed. 7 is the result of quantifying the weight ratio of acetone insolubles based on 100 g of dry raw material for the total phospholipid content of the egg yolk powder extract in which the purity of ethanol of Example 1-1 was changed to 70, 90, 95, 99.9% (w/w) .

As a result, as shown in FIG. 7 , it was found that the content of total phospholipids was highest at 95% (w/w) purity of ethanol. This means that when extracted using a mixed solvent of ethanol/medium-chain triglycerides, total phospholipids can be obtained in high yield at ethanol purity of 92 to 97% (w/w).

Experimental Example 4: Characteristics, phase separation observation, and active ingredient content analysis of ethanol/oil mixed solvent extract of egg yolk powder according to oil solvent

4.1 Characterization analysis

The following [Table 4] shows the results of confirming the properties of the ethanol/oil mixed solvent extract of the egg yolk powder of Example 1-3 and the oil solvent extract of the egg yolk powder of Comparative Examples 1-5, and FIG. 8 shows Examples 1-3 and Comparative Examples 1-5 This is a photograph confirming the properties of the extract.

extracted oil extraction temperature Properties of the mixed extract (Example 1-3) Properties of the oil extract (Comparative Example 1-5) egg yolk oil room temperature Yellow oil with a gel-like precipitate clear yellow oil Evening Primrose Oil room temperature Yellow oil with a gel-like precipitate clear yellow oil soybean oil room temperature Yellow oil with a gel-like precipitate yellow wax mineral oil room temperature uniform yellow oil clear yellow oil butter 50℃ Gel and yellow oil coexist yellow gel squalane room temperature uniform yellow oil clear yellow oil shea butter oil 50℃ yellow gel ivory-colored gel olive oil room temperature uniform yellow oil clear yellow oil coconut oil 50℃ Yellow oil with a gel-like precipitate Yellow oil with a gel-like precipitate palm oil 50℃ yellow gel bright yellow gel grapeseed oil room temperature uniform yellow oil clear yellow oil sunflower oil room temperature Yellow oil with a gel-like precipitate clear yellow oil jojoba oil room temperature uniform yellow oil clear yellow oil

As a result, as shown in [Table 4] and FIG. 8, the appearance of the ethanol/oil mixed extract of Example 1-3 had a precipitate or dark yellow, whereas the oil extract of Comparative Example 1-5 had a clear and bright color. extract came out. It existed in a liquid or solid state at room temperature depending on the melting point of the oil used.

4.2. Phase separation observation

The egg yolk powder extracts prepared in Examples 1-3 and Comparative Examples 1-5 were mixed with water in a weight ratio of 1:9, mixed with a hand mixer, and left at room temperature to observe phase separation with the naked eye.

As a result, phase separation was observed during extraction with the oil solvent of Comparative Example 1-5, but it was observed that the polar and amphoteric extracts of the ethanol layer stabilize the emulsion when extracted with the ethanol/oil mixed solvent of Examples 1-3.

9 is a photograph in which the butter extract prepared in Examples 1-3 and Comparative Example 1-5 in Experimental Example 4.2 was mixed with water in a weight ratio of 1:9, mixed with a hand mixer, and then visually observed whether or not phases were separated. . As a result, as shown in FIG. 9, the gel-like oil of Comparative Example 1-5 maintained the shape of a gel when mixed with water and was not mixed, but in the case of the gel-like oil of Example 1-2, it was easily mixed with water. confirmed to have That is, in the case of extraction with a mixed solvent, not only the non-polar material but also the amphoteric material serving as an emulsifier can be extracted to stabilize the emulsion, and it can be used to prepare a stable formulation for the manufacture of food and cosmetics.

4.3. Total phospholipid analysis

Total phospholipid analysis was performed in the same manner as in Experimental Example 2.2 on the ethanol/oil mixed extract of Examples 1-3 and the oil extract of Comparative Examples 1-5.

10 shows the total phospholipid content of the egg yolk powder extract extracted using the solvents of Examples 1-1 and 1-3 and the egg yolk powder extract extracted using the solvents of Comparative Examples 1-4 and 1-5 based on 100 g of acetone-insoluble This is the result of quantifying the weight ratio of water.

As a result, as shown in FIG. 10, it was confirmed that Examples 1-3 can obtain a total phospholipid (acetone insoluble) value that is 2.7 to 9.4 times higher than Comparative Examples 1-5.

4.4. Lutein Analysis

The egg yolk powder extracts prepared in Examples 1-3 and Comparative Examples 1-5 were analyzed according to the Chinese National Standards Method (GBT-23209-2008) lutein analysis method. Agilent, HPLC, and 1220 infinity were used as analytical instruments.

11 is a graph analyzing the lutein content of the egg yolk powder extract extracted using the solvents of Examples 1-1 and 1-3 and the egg yolk powder extract extracted using the solvents of Comparative Examples 1-4 and 1-5. 12 is a graph analyzing the lutein content of an egg yolk powder extract extracted using the solvent of Example 1-1 and an egg yolk powder extract extracted using the solvent of Comparative Examples 1-1 to 1-4.

As a result, as shown in Figures 11 and 12, Example 1-1 compared to Comparative Example 1-5, Example 1-3 compared to Comparative Example 1-5, the lutein content (ppm) was increased 1 to 6.0 times. . In addition, 9.3 ppm of Comparative Example 1-1, 8.3 ppm of Comparative Example 1-2, 4.4 ppm of Comparative Example 1-3, and 10.8 ppm of Comparative Example 1-4 were extracted. That is, compared to single extraction, lutein was efficiently extracted during mixed extraction.

Experimental Example 5. Phase separation observation and active ingredient content analysis of ethanol/medium chain neutral fat mixed solvent extracts of various natural products

5.1. character analysis

13 is a photograph confirming the properties of the mixed solvent extract of ethanol/medium chain neutral fat of various natural products of Example 2-1. 14 is a photograph confirming the properties of the ethanol solvent extract of various natural products of Comparative Example 2-2. As shown in FIGS. 13 and 14 , the mixed extract has fluidity, while the ethanol extract has no fluidity, and thus has an advantage in handling the extract.

5.2. Fatty Acid Analysis

Fatty acid analysis was performed in the same manner as in Experimental Example 2.1 for extracts of various natural products extracted using the solvents of Example 2-1 and Comparative Examples 2-1 and 2-2.

15 shows the fatty acid content of extracts of various natural products extracted using the ethanol/MCT mixed solvent of Example 2-1 and the extracts of various natural products extracted using the solvents of Comparative Examples 2-1 and 2-2 based on 100 g of dry raw material is the result of quantification.

As a result, as shown in FIG. 15, Example 2-1 extracted using an ethanol/MCT mixed solvent showed that 1 to 5.26 times more fatty acids were extracted than Comparative Examples 2-1 and 2-2, It was confirmed that fatty acids were extracted in an amount similar to the existing technology of extraction using Folch's solution, an organic solvent harmful to the human body. This means that when extracting using a mixed solvent of ethanol/medium-chain triglycerides, it is possible to extract fatty acids at a similar level to various natural products without using organic solvents harmful to the human body.

5.3. Total phospholipid analysis

Total phospholipid analysis was performed in the same manner as in Experimental Example 2.2 for extracts of various natural products extracted using the solvents of Example 2-1 and Comparative Example 2-2.

Figure 16 shows the total phospholipid content of various natural products extracted using the ethanol/MCT mixed solvent of Example 2-1 and the ethanol solvent of Comparative Example 2-2, the total phospholipid content of acetone insolubles based on 100 g of dry raw material. This is the result of quantifying the weight ratio.

As a result, as shown in FIG. 16 , Example 2-1 extracted using an ethanol/MCT mixed solvent obtained a similar amount of acetone insolubles as Comparative Example 2-2, which is an ethanol solvent extract. This means that MCT does not affect the solubility of polar substances soluble in ethanol.

5.4. Phase separation observation

The extracts prepared in Example 2-1 and Comparative Example 2-2 were mixed with water in a weight ratio of 1:9, mixed with a hand mixer, left at room temperature, and phase separation was observed with the naked eye and a microscope.

17 is a diagram showing a mixture of yolk powder, krill, buttermilk, egg yolk powder, and green-lipped mussel powder extracts extracted using the ethanol single solvent of Comparative Example 2-2 or the ethanol/MCT mixed solvent of Example 2-1 with water 1: This is a photograph of visual observation of phase separation after mixing with a hand mixer after mixing in a weight ratio of 9 (from left to right, green-lipped mussel powder extract, abalone yolk powder, krill shrimp, buttermilk, egg yolk powder, and green-lipped mussel powder extract). 18 is a diagram showing a mixture of green-lipped mussel powder extract with water in a weight ratio of 1:9, and then mixed with water and a hand mixer. It is the result of observing the phase separation under a microscope.

As a result, as shown in FIGS. 17 and 18 , the green-lipped mussel powder extract, krill, buttermilk, egg yolk, and phospholipids and neutral lipids were extracted using the ethanol/MCT mixed solvent of Example 2-1. It was extracted to form a stable emulsion upon mixing with water. Such an emulsion is stably maintained even after a month has elapsed after mixing, and the phospholipid component extracted in a polar solvent acts as an emulsifier to help disperse non-polar substances. In the case of extraction with a single solvent rather than mixed extraction, the upper color of the tube appeared dark, phase separation was observed, and no emulsion was formed.

5.5 Total Polyphenol Analysis

Total polyphenol analysis was performed on the extracts of evening primrose seeds extracted using the solvents of Example 2-2 and Comparative Example 2-4. The total polyphenol analysis was conducted by the Functional Food Division of the National Academy of Agricultural Science, except that the dilution solvent was used as 50% (w/w) ethanol. PJ006781200902) was analyzed according to the analysis method. As an analysis instrument, PG instruments Ltd, UV-Spectrometer, and T60U Spectrometer were used.

19 shows the total polyphenol content of the evening primrose seed extract extracted using the ethanol/water/MCT mixed solvent of Example 2-2 and the evening primrose seed extract extracted using the ethanol/water solvent of Comparative Example 2-4 as a dry raw material It is the result of quantification based on 100g.

As a result, as shown in FIG. 19, Example 2-2 extracted using an ethanol/water/MCT mixed solvent obtained a total polyphenol analysis value similar to that of Comparative Example 2-4, an ethanol/water solvent extract, It was confirmed that more total polyphenols could be obtained when the purity of ethanol was 50% (w/w).

5.6 DPPH radical scavenging activity analysis

DPPH radical scavenging activity analysis was performed on the extracts of evening primrose seeds extracted using the solvents of Example 2-2 and Comparative Example 2-4. Specifically, a sample in which the ethanol/water layer was separated from the extract of evening primrose seeds extracted using a 50:50 weight ratio of ethanol and water in the ethanol/water/MCT mixed solvent of Example 2-2 (month 50:50, ethanol /water layer) and samples including both ethanol/water layer and MCT layer (month 50:50 total extract), evening primrose seeds extracted using a 50:50 weight ratio of ethanol and water in the ethanol/water solvent of Comparative Example 2-4 Antioxidant scavenging activity was analyzed for extracts of (1:4:0, 50%). DPPH radical scavenging activity was analyzed according to the analysis method (DPPH antioxidant assay revisited. Food Chemistry 2009;113:1202-1205), except that the dilution solvent was 99.9% (w/w) ethanol. As an analysis instrument, PG instruments Ltd, UV-Spectrometer, and T60U Spectrometer were used.

20 is an extract of evening primrose seeds extracted using the ethanol/water/MCT mixed solvent of Example 2-4 (month 50:50 ethanol/water layer, moon 50:50 total extract), ethanol/water of Comparative Example 2-4 50% inhibitory concentration (IC ₅₀ ) of evening primrose seed extract extracted using solvent, tocopherol (JB, Mixed Tocopherol T-70), and evening primrose oil extracted by cold pressing from evening primrose seed oil (Saero Hands, evening primrose oil tablet) As a result of quantification in ppm, FIG. 20A is a general graph, and FIG. 20B is a log graph.

As a result, as shown in FIG. 20, Example 2-2 (month 50:50, ethanol/water layer) extracted using an ethanol/water/MCT mixed solvent was an ethanol/water solvent extract, Comparative Example 2-4 (1: 4:0, 50%) showed a statistically similar IC ₅₀ value, and it was found to have about 3 times the antioxidant scavenging activity than tocopherol. In addition, Example 2-2 (month 50:50, ethanol/water layer) has about 3000 times the antioxidant scavenging activity of evening primrose oil extracted from oil by cold pressing from evening primrose seeds, and Example 2-2 (month 50:50) total extract) was also confirmed to have about 100 times more antioxidant scavenging activity than evening primrose oil. This means that the antioxidants contained in evening primrose seeds can be extracted at a high concentration when extracted using an ethanol/MCT mixed solvent.

5.7 Ginsenoside analysis

Ginsenoside analysis was performed on the extract of red ginseng extracted using the solvent of Example 2-2 and Comparative Example 2-4. Ginsenoside analysis was performed according to the Food Standards Code 6.4.2.1 Ginseng/Red Ginseng Ingredients (as ginsenosides). Agilent, HPLC, and 1220 infinity were used as analytical instruments.

21 shows the ginsenoside content of the ethanol/water layer of the extract of red ginseng extracted using the ethanol/water/MCT mixed solvent of Example 2-2 and the extract of red ginseng extracted using the ethanol/water solvent of Comparative Example 2-4. is the result of quantification based on 1 g of dry raw material.

As a result, as shown in FIG. 21, Example 2-2 extracted using an ethanol/water/MCT mixed solvent obtained similar results to Comparative Example 2-4, which is an ethanol/water solvent extract. This means that MCT does not affect the solubility of polar substances soluble in ethanol.

Claims (16)

(A) extracting by adding a polar solvent and a non-polar solvent to a natural product and stirring;
(B) filtering the extract to remove solids to obtain a filtrate; and
(C) forming an immiscible non-polar solvent layer and a polar solvent layer by allowing the obtained filtrate to stand still; a method of simultaneously extracting a polar component and a non-polar component from a natural product using an immiscible solvent comprising a. The method according to claim 1, After step (C),
obtaining an extract containing a non-polar component by separating the formed non-polar solvent layer and the polar solvent layer; and
The method further comprising; evaporating the polar solvent from the separated polar solvent layer to obtain an extract containing a polar component. The method according to claim 1, After step (C),
The method further comprising; evaporating the polar solvent without separating the non-polar solvent layer and the polar solvent layer to obtain an extract containing a non-polar component and a polar component. The method according to claim 1, wherein the polar solvent is a C1-C6 alcohol, water, acetone, ketone, or a mixture thereof. The method according to claim 4, wherein the C1-C6 alcohol is methanol, ethanol, n-propanol, isopropanol, n-butanol, sec-butanol, isobutanol, tert-butanol, n-pentanol, n-hexanol, 1,2 -Hexanediol, 1,6-hexanediol, or a mixture thereof. The method according to claim 1, wherein the non-polar solvent is medium chain triglycerides (MCT), egg yolk oil, evening primrose oil, soybean oil, mineral oil, butter, squalane, shea butter oil, olive oil, coconut oil, palm oil, grape seed oil, sunflower oil, jojoba oil, or a mixture thereof. The method according to claim 1, wherein the natural product is selected from the group consisting of egg yolk powder, yellow yolk powder, soybean, green-lipped mussel, krill, buttermilk powder, green bean coffee, cheonggukjang, rice bran, evening primrose seed, ginseng, and red ginseng. How to be either. The method according to claim 1, wherein the non-polar component is a method comprising at least one selected from the group consisting of triglycerides, carotenoids, and lutein. The method according to claim 1, wherein the polar component comprises at least one selected from the group consisting of phospholipids, polyphenols, and ginsenosides. The method according to claim 1, wherein the polar solvent has a purity of 20 to 99.9% (w/w). The method according to claim 1, wherein step (A) is based on 100 parts by weight of the natural product, 100 to 800 parts by weight of a polar solvent, and 10 to 1600 parts by weight of a non-polar solvent. A natural product extract containing a non-polar component, a polar component, or a combination thereof prepared by the method according to any one of claims 1 to 11. The natural extract according to claim 12, wherein the non-polar component comprises at least one selected from the group consisting of neutral lipids, carotenoids, and lutein. The natural product extract of claim 12, wherein the polar component comprises at least one selected from the group consisting of phospholipids, polyphenols, and ginsenosides. A food composition comprising a natural product extract containing the non-polar component of claim 12, the polar component, or a combination thereof. A cosmetic composition comprising a natural extract containing the non-polar component of claim 12, the polar component, or a combination thereof.

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