Natural hydrogen bond donor solvent and method for extracting active ingredients using same

Abstract

The present invention relates to a natural hydrogen bond donor solvent and a method for extracting active ingredients using same, wherein the natural hydrogen bond donor solvent is prepared by mixing glucose and lactic acid. The natural hydrogen bond donor has an excellent effect of extracting active ingredients from an extraction target and is expected to discharge fewer environmental pollutants due to the absence of toxic substance emission and by-products. Additionally, physiologically active ingredients can be extracted with high efficiency from various plants.

Description

Natural hydrogen bond donor solvent and method for extracting effective ingredients using the same

[0001] The present invention relates to a natural hydrogen bond donor solvent and a method for extracting an effective ingredient using the same.

[0002] The existing industrial extraction methods for plant components such as natural products or herbal medicines can be broadly divided into high-temperature hot water extraction and organic solvent extraction.

[0003] Hot water extraction offers the advantage of extracting a variety of water-soluble substances and is the most widely used method because it eliminates the need for residual toxicity of the extraction solvent. However, hot water extraction is difficult to extract from nonpolar substances, has problems such as heat-induced decomposition or denaturation of active ingredients, and requires additional post-extraction processes such as freeze-drying.

[0004] In addition, extraction methods using organic solvents have advantages over hot water extraction methods in extracting non-polar substances, but they have limitations such as the possibility of causing cytotoxicity and skin irritation due to residual solvents, so a secondary solvent removal process is essential after the extraction process.

[0005] To overcome the shortcomings of these hydrothermal and organic solvent extraction methods, deep eutectic solvents (DES) have been developed as an environmentally friendly alternative. DES possesses properties such as high solubility and a low melting point, and has the advantages of low toxicity, low cost, and easy and simple production.

[0006] The eutectic solvent used in the present invention is a natural hydrogen bond donor solvent manufactured by mixing glucose and lactic acid. Unlike existing eutectic solvents, it is a solvent manufactured using naturally occurring substances, glucose and lactic acid. Glucose is mainly produced during photosynthesis or cellular respiration, and lactic acid is generally produced during fermentation by bacteria or other microorganisms. Glucose and lactic acid play different roles in different metabolic pathways in the human body and are known to be substances that contribute to energy production and regulation in the human body. This eco-friendly energy solvent is manufactured using biologically derived and inexpensive materials, making it environmentally friendly and applicable as a biocompatible solvent.

[0007] Accordingly, the present inventors have developed a natural hydrogen bond donor solvent that is low in toxicity, inexpensive, and can be manufactured using an easy and simple method. Using this natural hydrogen bond donor solvent, they attempted to extract effective ingredients, and in particular, to produce plant extracts with increased physiologically active ingredients. They confirmed that these plant extracts had increased total phenol, total flavonoid, and tannin contents, thereby completing the present invention.

[0008]

[0009] Prior art document: Republic of Korea Patent No. 10-2276980

[0010] An object of the present invention is to provide a natural hydrogen bond donor solvent (NHBDS) in which two or more hydrogen bond donor (HBD) solvents are mixed.

[0011] In addition, another object of the present invention is to provide a method for producing a natural hydrogen bond donor solvent (NHBDS).

[0012] In addition, another object of the present invention is to provide a method for extracting useful components.

[0013] In addition, another object of the present invention is to provide a method for increasing the content of physiologically active ingredients in a plant extract.

[0014] To achieve the above-described purpose, the present invention provides a natural hydrogen bond donor solvent (NHBDS) comprising two or more hydrogen bond donor (HBD) solvents.

[0015] Next, the present invention provides a method for producing a natural hydrogen bond donor solvent (NHBDS), comprising a step of mixing two or more hydrogen bond donor (HBD) solvents.

[0016] Furthermore, the present invention provides a method for extracting a useful component, including a step of extracting a useful component from animals, plants, prokaryotes, and eukaryotes using a natural hydrogen bond donor solvent in which two or more types of hydrogen bond donor solvents are mixed.

[0017] Lastly, the present invention provides a method for increasing the content of a physiologically active component in a plant extract, comprising the steps of: preparing a plant; mixing two or more hydrogen bond donor solvents to prepare a natural hydrogen bond donor solvent (NHBDS); and adding the natural hydrogen bond donor solvent to the plant to extract it.

[0018] The present invention produces a natural hydrogen bond donor solvent mixed with glucose and lactic acid. This natural hydrogen bond donor is highly effective in extracting active ingredients from the target substance, and is expected to produce minimal environmental pollutants due to its absence of toxic substances and byproducts. Furthermore, it can efficiently extract physiologically active ingredients from various plants.

[0019] FIG. 1 is a diagram showing the results of measuring the total phenol content of a Platycodon grandiflorum extract in one embodiment of the present invention.

[0020] Figure 2 is a diagram showing the results of measuring the total phenol content of a bottle gourd extract in one embodiment of the present invention.

[0021] Figure 3 is a diagram showing the results of measuring the total phenol content of a ginseng extract in one embodiment of the present invention.

[0022] Figure 4 is a diagram showing the results of measuring the total phenol content of a licorice extract in one embodiment of the present invention.

[0023] FIG. 5 is a diagram showing the results of measuring the total phenol content of an extract of Angelica gigas Nakai in one embodiment of the present invention.

[0024] Figure 6 is a diagram showing the results of measuring the total phenol content of an extract of Schisandra chinensis according to one embodiment of the present invention.

[0025] Figure 7 is a diagram showing the results of measuring the total phenol content of a green tea extract in one embodiment of the present invention.

[0026] Figure 8 is a diagram showing the results of measuring the total phenol content of a gold extract in one embodiment of the present invention.

[0027] FIG. 9 is a diagram showing the results of measuring the total flavonoid content of a Platycodon grandiflorum extract in one embodiment of the present invention.

[0028] Figure 10 is a diagram showing the results of measuring the total flavonoid content of a bottle gourd extract in one embodiment of the present invention.

[0029] Figure 11 is a diagram showing the results of measuring the total flavonoid content of a ginseng extract in one embodiment of the present invention.

[0030] Figure 12 is a diagram showing the results of measuring the total flavonoid content of a licorice extract in one embodiment of the present invention.

[0031] Figure 13 is a diagram showing the results of measuring the total flavonoid content of the extract of Angelica gigas Nakai in one embodiment of the present invention.

[0032] Figure 14 is a diagram showing the results of measuring the total flavonoid content of an extract of Schisandra chinensis according to one embodiment of the present invention.

[0033] Figure 15 is a diagram showing the results of measuring the total flavonoid content of a green tea extract in one embodiment of the present invention.

[0034] Figure 16 is a diagram showing the results of measuring the total flavonoid content of a gold extract in one embodiment of the present invention.

[0035] Figure 17 is a diagram showing the results of measuring the condensed tannin content of a Platycodon grandiflorum extract in one embodiment of the present invention.

[0036] Figure 18 is a diagram showing the results of measuring the condensed tannin content of a bottle gourd extract in one embodiment of the present invention.

[0037] Figure 19 is a diagram showing the results of measuring the condensed tannin content of a ginseng extract in one embodiment of the present invention.

[0038] Figure 20 is a diagram showing the results of measuring the condensed tannin content of a licorice extract in one embodiment of the present invention.

[0039] Figure 21 is a diagram showing the results of measuring the condensed tannin content of the extract of Angelica gigas Nakai in one embodiment of the present invention.

[0040] Figure 22 is a diagram showing the results of measuring the condensed tannin content of an extract of Schisandra chinensis according to one embodiment of the present invention.

[0041] Figure 23 is a diagram showing the results of measuring the condensed tannin content of a green tea extract in one embodiment of the present invention.

[0042] Figure 24 is a diagram showing the results of measuring the condensed tannin content of a golden extract in one embodiment of the present invention.

[0043] Figure 25 is a diagram showing the results of measuring the hydrolyzed tannin content of a doraji extract in one embodiment of the present invention.

[0044] Figure 26 is a diagram showing the results of measuring the hydrolyzed tannin content of a bottle gourd extract in one embodiment of the present invention.

[0045] Figure 27 is a diagram showing the results of measuring the hydrolyzed tannin content of a ginseng extract in one embodiment of the present invention.

[0046] Figure 28 is a diagram showing the results of measuring the hydrolyzed tannin content of a licorice extract in one embodiment of the present invention.

[0047] Figure 29 is a diagram showing the results of measuring the hydrolyzed tannin content of an extract of Angelica gigas Nakai in one embodiment of the present invention.

[0048] Figure 30 is a diagram showing the results of measuring the hydrolyzed tannin content of an extract of Schisandra chinensis according to one embodiment of the present invention.

[0049] Figure 31 is a diagram showing the results of measuring the hydrolyzed tannin content of a green tea extract in one embodiment of the present invention.

[0050] Figure 32 is a diagram showing the results of measuring the hydrolyzed tannin content of a golden extract in one embodiment of the present invention.

[0051] Figure 33 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of a Platycodon grandiflorum extract in one embodiment of the present invention.

[0052] Figure 34 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of the extract of the purslane plant in one embodiment of the present invention.

[0053] Figure 35 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of a ginseng extract in one embodiment of the present invention.

[0054] Figure 36 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of a licorice extract in one embodiment of the present invention.

[0055] Figure 37 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of the extract of Angelica gigas Nakai in one embodiment of the present invention.

[0056] Figure 38 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of an extract of Schisandra chinensis according to one embodiment of the present invention.

[0057] Figure 39 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of a green tea extract in one embodiment of the present invention.

[0058] Figure 40 is a diagram showing the results of confirming the HPLC (High-performance liquid chromatography) spectrum of a gold extract in one embodiment of the present invention.

[0059] Hereinafter, the present invention will be described in detail with reference to the attached drawings, using exemplary embodiments. However, the following exemplary embodiments are provided as illustrative examples of the present invention. If a detailed description of a technology or configuration well known to those skilled in the art is judged to unnecessarily obscure the gist of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the following claims and equivalents interpreted therefrom.

[0060] In addition, the terms used in this specification are terms used to appropriately express preferred embodiments of the present invention, and may vary depending on the intention of the user or operator, or the customs of the field to which the present invention belongs. Therefore, the definitions of these terms should be determined based on the contents throughout this specification. Throughout the specification, when a part is said to "include" a certain component, this does not mean that other components are excluded, but rather that other components may be included, unless specifically stated otherwise.

[0061] Throughout this specification, '%' used to indicate the concentration of a particular substance is (w / w) % for solid / solid, (w / v) % for solid / liquid, and (v / v) % for liquid / liquid, unless otherwise stated.

[0062] Hereinafter, the present invention will be described in more detail.

[0063]

[0064] In one aspect, the present invention provides a natural hydrogen bond donor solvent (NHBDS) comprising two or more hydrogen bond donor (HBD) solvents mixed together.

[0065] In one embodiment of the present invention, the hydrogen bond donor solvent may be selected from the group consisting of monosaccharides, disaccharides, polysaccharides, amino acids, carboxylic acids, and aldehydes, preferably polysaccharides and carboxylsal, more preferably glucose and lactic acid, but is not limited thereto.

[0066] In one embodiment of the present invention, when two types of the hydrogen bond donor solvents are selected and used, the hydrogen bond donor solvents may be mixed in a molar ratio of 1 to 3:1 to 5, preferably 1 to 2:1 to 4, but is not limited thereto.

[0067] In one embodiment of the present invention, the hydrogen bond donor solvent may be a mixture of at least one natural hydrogen bond donor solvent selected from monosaccharides, disaccharides, and polysaccharides, preferably polysaccharides, more preferably glucose; and at least one natural hydrogen bond donor solvent selected from amino acids, carboxylic acids, and aldehydes, preferably carboxylic acids, more preferably lactic acid; but is not limited thereto.

[0068] In the present invention, the glucose has an aldehyde group (-CHO) that can act as a hydrogen donor and a hydroxyl group (-OH) that can act as a hydrogen donor, and the lactic acid has a carboxyl group (-COOH) that can act as a hydrogen donor and a hydroxyl group (-OH) that can act as a hydrogen donor. Therefore, glucose and lactic acid can act as hydrogen donors to form hydrogen bonds, and in the present invention, a new type of solvent was prepared by combining the glucose and lactic acid, and this was named a natural hydrogen bond donor solvent.

[0069] The above natural hydrogen bond donor solvent can be used directly as an extraction solvent, but when the viscosity is high, it can be mixed with another solvent to increase extraction efficiency, and preferably, it can be used as a solvent by mixing with water.

[0070] The above water may be added to the natural hydrogen bond donor solvent in a volume ratio of 0.1 to 0.3: 1 to 2, but is not limited thereto.

[0071]

[0072] In one aspect, the present invention provides a method for producing a natural hydrogen bond donor solvent (NHBDS), comprising the step of mixing two or more hydrogen bond donor (HBD) solvents.

[0073] In one embodiment of the present invention, when two types of the hydrogen bond donor solvents are selected and used, the hydrogen bond donor solvents may be mixed in a molar ratio of 1 to 3:1 to 5, preferably 1 to 2:1 to 4, but is not limited thereto.

[0074]

[0075] In one aspect, the present invention provides a method for extracting a useful component, comprising the step of extracting a useful component from animals, plants, prokaryotes, and eukaryotes using a natural hydrogen bond donor solvent in which two or more hydrogen bond donor solvents are mixed.

[0076] In one embodiment of the present invention, the plant may be at least one selected from the group consisting of Platycodon grandifloras, Centella asiatica, Panax ginseng, Glycyrrhiza uralensis, Angelica gigas, Schisandra chinensis, Camellia sinensis, and Scutellaria baicalensis, but is not limited thereto.

[0077]

[0078] In one aspect, the present invention relates to a method for increasing the content of a physiologically active component in a plant extract, comprising the steps of: preparing a plant; mixing two or more hydrogen bond donor solvents to prepare a natural hydrogen bond donor solvent (NHBDS); and adding the natural hydrogen bond donor solvent to the plant to extract it.

[0079] In one embodiment of the present invention, the plant may be at least one selected from the group consisting of, but is not limited to, Platycodon grandiflorum, Centella asiatica, ginseng, licorice, Angelica gigas, Schisandra chinensis, green tea, and goldenrod.

[0080] In one embodiment of the present invention, the natural hydrogen bond donor solvent may be, but is not limited to, an ultrasonic-treated solvent.

[0081] In one embodiment of the present invention, the ultrasonic treatment may be performed for 20 to 40 minutes, preferably 25 to 35 minutes, and more preferably 30 minutes, but is not limited thereto.

[0082] In one embodiment of the present invention, the natural hydrogen bond donor solvent may be added in an amount of 8 to 12 times the weight of the plant body, preferably 9 to 11 times, and more preferably 10 times, but is not limited thereto.

[0083] In one embodiment of the present invention, the extraction may be performed at 30 to 70°C, preferably 40 to 60°C, more preferably 50°C, for 20 to 40 minutes, preferably 25 to 35 minutes, more preferably 30 minutes, but is not limited thereto.

[0084] In one embodiment of the present invention, the physiologically active ingredient may be at least one selected from the group consisting of total phenol, total flavonoid, tannin, platycodin, madecassoside, asiaticoside, ginsenoside, glycyrhizin, nodakenin, gomisin A, epicatechin, baicalin, baicalein, and wogonin, but is not limited thereto.

[0085] In one embodiment of the present invention, the platycodin may be an effective ingredient of a Platycodin extract, but is not limited thereto.

[0086] In one embodiment of the present invention, the madecassoside or asiaticoside may be an effective ingredient of the extract of Centella asiatica, but is not limited thereto.

[0087] In one embodiment of the present invention, the ginsenoside may be an effective ingredient of a ginseng extract, but is not limited thereto.

[0088] In one embodiment of the present invention, the ginsenoside may be at least one selected from the group consisting of Rg1, Rb1, and Rg3, but is not limited thereto.

[0089] In one embodiment of the present invention, the glycyrrhizin may be an effective ingredient of a licorice extract, but is not limited thereto.

[0090] In one embodiment of the present invention, the Nodakenin may be an effective ingredient of an extract of Angelica gigas Nakai, but is not limited thereto.

[0091] In one embodiment of the present invention, the Gomisin A may be an effective ingredient of an extract of Schisandra chinensis, but is not limited thereto.

[0092] In one embodiment of the present invention, the epicatechin may be an effective ingredient of green tea extract, but is not limited thereto.

[0093] In one embodiment of the present invention, the baicalin, baicalein or wogonin may be an effective ingredient of the golden extract, but is not limited thereto.

[0094]

[0095] This invention was carried out with the support of the Regional Research and Development Innovation Support Project (2023 KNU-DGG BIZ Linked Open Innovation Lab Support Project) of the Ministry of Science and ICT (Science and Technology Promotion Agency) with the project identification number 202322260300 and the project detailed number 2023-06.

[0096] As described above, specific embodiments of the present invention have been described in detail. However, those skilled in the art who understand the spirit of the present invention will be able to easily suggest other inventions that are backwards or other embodiments included within the scope of the spirit of the present invention by adding, changing, or deleting other components within the scope of the same spirit. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention is indicated by the scope of the claims described below rather than the detailed description described above, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

[0097]

[0098] <Example 1> Plant preparation

[0099] Platycodon grandifloras (Jecheon Herb, Jecheon-si, Chungcheongbuk-do), Centella asiatica (Jecheon Herb, Jecheon-si, Chungcheongbuk-do), Panax ginseng (Geumsan Fresh Ginseng Farming Association, Geumsan-gun, Chungcheongnam-do), Glycyrrhiza (Glycyrrhiza uralensis) (Jecheon Herb, Jecheon-si, Chungcheongbuk-do), Angelica gigas (Pyeongchang-gun, Gangwon-do), Schisandra chinensis (Schisandra chinensis) (Jecheon Herb, Jecheon-si, Chungcheongbuk-do), Green tea (Camellia sinensis) (Jecheon Herb, Jecheon-si, Chungcheongbuk-do), and Scutellaria baicalensis (Jecheon Medicinal Herb Farming Association, Jecheon-si, Chungcheongbuk-do) were dried, ground, and used.

[0100]

[0101] <Example 2> Preparation of Natural Hydrogen Bond Donor Solvent (NHBDS)

[0102] In order to prepare a natural hydrogen bond donor solvent, hydrogen bond donor solvents, glucose and lactic acid, were mixed at a molecular ratio as shown in Table 1 below. After mixing, 20 mL (v / v) of water was added to the mixture, and the ultrasonic power was set to 50%, followed by ultrasonic treatment for 30 minutes to prepare a natural hydrogen bond donor solvent.

[0103] Hydrogen bond donor 1 Hydrogen bond donor 2 Hydrogen bond donor 3 G1L1GlucoseLactic acid 1:1 G1L2GlucoseLactic acid 1:2 G1L3GlucoseLactic acid 1:3 G1L4GlucoseLactic acid 1:4 G2L3GlucoseLactic acid 2:3

[0104] <Example 3> Preparation of plant extracts

[0105] To prepare a plant extract, 10 mL of the natural hydrogen bond donor solvent prepared in Example 2 was added to 1 g of the plant prepared in Example 1, and extraction was performed at 50°C for 30 minutes to prepare an extract. In addition, 70% ethanol, an organic solvent, was used as a control, and extraction was performed under the same conditions.

[0106]

[0107] <Experimental Example 1> Measurement of total phenol and total flavonoid content

[0108] 1-1. Measurement of total phenol content

[0109] Total phenol contents (TPC) were measured in the plant extract prepared in Example 3 above.

[0110] Total phenol content was determined by adding 2 μl of plant extract, 20 μl of distilled water, and 10 μl of Folin-Ciocalteu's phenol reagent to a 96-well plate and incubating at room temperature for 6 minutes. Then, 100 μl of 7% Na2CO3 solution and 70 μl of distilled water were added and incubated at room temperature for 90 minutes, and the absorbance was measured at 595 nm. A calibration curve was created using gallic acid as a standard, and the total phenol content measured using this is shown in Figures 1 to 8.

[0111] As shown in Figures 1 to 8, the total phenol content was significantly higher in plant extracts extracted with a natural hydrogen bond donor solvent than in plant extracts extracted with ethanol.

[0112]

[0113] 1-2. Measurement of total flavonoid content

[0114] Total flavonoid contents (TFC) were measured in the plant extract prepared in Example 3 above.

[0115] Total flavonoid content was determined by adding 2 μl of plant extract, 100 μl of distilled water, and 5 μl of 5% NaNO2 solution to a 96-well plate and incubating at room temperature for 10 minutes. Then, 10 μl of 10% AlCl3·6H2O solution was added and incubated again at room temperature for 10 minutes. After adding 40 μl of 1 M NaOH and 45 μl of distilled water, the absorbance was measured at 405 nm. A calibration curve was created using catechin as a standard, and the total flavonoid content measured using this is shown in Figures 9 to 16.

[0116] As shown in Figures 9 to 16, the total flavonoid content was significantly higher in plant extracts extracted with a natural hydrogen bond donor solvent than in plant extracts extracted with ethanol.

[0117] Through this, it was confirmed that the total phenol content and total flavonoid content of the plant extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid were significantly high, and accordingly, it was judged that a natural hydrogen bond donor solvent mixed with glucose and lactic acid could be used as a solvent to replace organic solvents.

[0118]

[0119] <Experimental Example 2> Tannin content analysis

[0120] 2-1. Analysis of condensed tannin content

[0121] The content of condensed tannin, a flavonoid compound, was measured in the plant extract prepared in Example 3 above.

[0122] The condensed tannin content was measured by adding 2 μl of plant extract, 100 μl of 1% vanillin, and 100 μl of 25% H2SO4 to a 96-well plate, incubating at room temperature for 10 minutes, and measuring the absorbance at 500 nm. A calibration curve was created using catechin as a standard, and the condensed tannin content measured using this is shown in Figures 17 to 24.

[0123] As shown in Figures 17 to 24, the condensed tannin content was significantly higher in plant extracts extracted with a natural hydrogen bond donor solvent than in plant extracts extracted with ethanol.

[0124]

[0125] 2-2. Analysis of hydrolyzed tannin content

[0126] The content of hydrolysable tannin, a flavonoid compound, was measured in the plant extract prepared in Example 3 above.

[0127] For hydrolyzed tannin analysis, 2 μl of plant extract, 100 μl of DW, and 100 μl of 2.5% KIO3 were added to a 96-well plate, incubated at room temperature for 5 minutes, and the absorbance was measured at 550 nm. Tannic acid was used as a standard substance, and the hydrolyzed tannin content measured using this is shown in Figures 25 to 32.

[0128] As shown in Figures 25 to 32, the content of hydrolyzed tannins was significantly higher in plant extracts extracted with a natural hydrogen bond donor solvent than in plant extracts extracted with ethanol.

[0129] Through this, it was confirmed that the tannin content of plant extracts extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid was significantly high and possessed antioxidant activity. Accordingly, it was determined that a natural hydrogen bond donor solvent mixed with glucose and lactic acid could be used as a solvent to replace organic solvents.

[0130]

[0131] <Experimental Example 3> Analysis of the content of unique physiologically active components contained in plant extracts

[0132] 3-1. Analysis of platycodin content in Platycodin extract

[0133] The content of platycodin included in the control group, the ethanol extract of Platycodon grandiflorum prepared in Example 3 above, and the G1L3 group extracted with the G1L3 eutectic solvent was measured.

[0134] To measure the platycodin content, 2 μl of Platycodon grandiflorum extract, 100 μl of DW, and 100 μl of 2.5% KIO3 were sequentially added and incubated at room temperature for 5 minutes. HPLC (High-performance liquid chromatography) was measured, and the peak area at the retention time (RT) corresponding to each active ingredient was calculated to compare the active ingredient content. The analysis conditions for HPLC are shown in Table 2 below.

[0135] Column 4.6×150 mm, 5 μm, C18 Mobile Phase Distilled water:acetonitrile (A:B) Flow rate 0.9 mL / min Analysis time 75 min Wavelength 205 nm, ESLD Temperature 30 ℃ Injection volume 10 ㎕

[0136] As a result, as shown in Table 3, Table 4 and Figure 33 below, it was confirmed that the platycodin content of the G1L3 group of the Platycodon grandiflorum extract using the natural hydrogen bond donor solvent of the present invention was significantly increased compared to the control group, which was the ethanol extract group.

[0137] Specifically, in the HPLC spectrum, at a retention time (RT) of 27.0 minutes corresponding to platycodin, the peak area of ​​the G1L3 group increased approximately 3 times compared to the control group.

[0138] Active ingredient R T Peak area Height Conc. Unit Control group Platycodin 27.049355647428850.063 mM

[0139]

[0140] Active ingredient RTPeak areaHeightConc.UnitG1L3Platicodine27.059989035955710.202mM

[0141] Through this, it was confirmed that the platycodin content of the Platycodon extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid was significantly higher than that of the control group.

[0142]

[0143] 3-2. Analysis of Madecassoside and Asiaticoside Contents in Centella Asiatica Extract

[0144] The contents of Madecassoside and Asiaticoside included in the control group, the ethanol extract of Centella asiatica prepared in Example 3, and the G1L3 group extracted with the G1L3 eutectic solvent were measured.

[0145] The process for measuring the contents of madecassoside and asiaticoside was carried out in the same manner as in Experimental Example 3-1 above, and the analysis conditions of HPLC are shown in Table 5 below.

[0146] Column 4.6×250 mm, 5 μm, C18 Mobile Phase Distilled water:acetonitrile (A:B) Flow rate 1.0 mL / min Analysis time 55 min Wavelength 206 nm Temperature 30 ℃ Injection volume 20 ㎕

[0147]

[0148] As a result, as shown in Table 6, Table 7 and Figure 34 below, compared to the control group, which is an ethanol extract group, it was confirmed that the contents of madecassoside and asiaticoside of the G1L3 group of the Centella asiatica extract using the natural hydrogen bond donor solvent of the present invention were significantly increased compared to the control group.

[0149] Specifically, in the HPLC spectrum, at a retention time (RT) of 14.9 minutes corresponding to madecassoside, the peak area of ​​the G1L3 group increased approximately 4.4 times compared to the control group, and at a retention time of 17.3 minutes corresponding to asiaticoside, the peak area of ​​the G1L3 group increased approximately 3.7 times compared to the control group.

[0150]

[0151] Active ingredient R T Peak area Height Concentration Unit Control Madecassoside 14.9 2 9 3 5 4 0 3 4 6 7 2 4 3 0 8.90 1 mg / L Asiaticoside 17.3 1 7 8 2 4 0 7 9 3 0 1 9 3 1 7.39 7 mg / L

[0152]

[0153] Active ingredient R T Peak area Height Conc. Unit G 1 L 3 Madecassoside 14.9 4 5 15 6 4 7 19 2 19 6 9 9 13 6 9.270 mg / L Asiaticoside 17.3 3 5 2 8 9 3 8 7 7 3 8 3 4 12 1 1 7 6.285 mg / L

[0154] Through this, it was confirmed that the contents of madecassoside and asiaticoside in the extract of Centella asiatica extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid were significantly higher than those in the control group.

[0155]

[0156] 3-3. Analysis of ginsenoside content in ginseng extracts

[0157] The ginsenoside content included in the ginseng ethanol extract, which was the control group prepared in Example 3, and the G2L3 group extracted with the G2L3 eutectic solvent was measured.

[0158] The process for measuring the contents of ginsenosides Rg1, Rb1, and Rg3 was carried out in the same manner as in Experimental Example 3-1, and the analysis conditions of HPLC are shown in Table 8 below.

[0159] Column 4.6×150 mm, 5 μm, C18 Mobile Phase Distilled water:acetonitrile (A:B) Flow rate 0.9 mL / min Analysis time 75 min Wavelength 205 nm, ESLD Temperature 30 ℃ Injection volume 10 ㎕

[0160] As a result, as shown in Table 8, Table 9 and Figure 35 below, it was confirmed that the contents of ginsenosides Rg1, Rb1 and Rg3 of the ginseng extract G2L3 group using the natural hydrogen bond donor solvent of the present invention were significantly increased compared to the control group, which was the ethanol extract group.

[0161] Specifically, in the HPLC spectrum, at a retention time (RT) of 25.0 minutes corresponding to Rg1, the peak area of ​​the G2L3 group increased by more than 3 times compared to the control group. At a retention time of 29.6 minutes corresponding to Rb1, the peak area of ​​the G2L3 group increased by more than 5.5 times compared to the ethanol extract group. At a retention time of 39.6 minutes corresponding to Rg3, the peak area of ​​the ethanol extract group did not appear, but the peak area of ​​the G2L3 group appeared large.

[0162] Active ingredient RTPeak areaHeightConc.UnitControl Rg124.950539236608290.170mMRb129.626132037146200.062mMRg3Not measured

[0163]

[0164] Active ingredient RTPeak areaHeightConc.UnitG2L3GroupRg124.93217204981737000.487mMRb129.605738845690240.336mMRg339.621403475197690.163mM

[0165] Through this, it was confirmed that the ginsenoside content of the ginseng extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid was significantly higher than that of the control group.

[0166]

[0167] 3-4. Analysis of glycyrrhizin content in licorice extract

[0168] The content of glycyrrhizin included in the control group, licorice ethanol extract prepared in Example 3 above, and the G1L4 group extracted with the G1L4 eutectic solvent was measured.

[0169] The process for measuring glycyrrhizin content was carried out in the same manner as in Experimental Example 3-1 above, and the HPLC analysis conditions are shown in Table 11 below.

[0170] Column 4.6×250 mm, 5 μm, C18 Mobile Phase Distilled water:acetonitrile (A:B) Flow rate 1 mL / min Analysis time 55 min Wavelength 206 nm Temperature 30 ℃ Injection volume 20 ㎕

[0171] As a result, as shown in Table 12, Table 13 and Figure 36 below, it was confirmed that the glycyrrhizin content of the licorice extract G1L4 group using the natural hydrogen bond donor solvent of the present invention was significantly increased compared to the control group, which was the ethanol extract group.

[0172] Specifically, in the HPLC spectrum, the peak area of ​​the G1L4 group at the retention time (RT) corresponding to glycyrrhizin increased approximately 26-fold compared to the control group.

[0173] Active ingredient R T Peak area Height Concentration Unit Control group Glycyrrhizin 36.94 256 340 352 939 32.648 mg / L

[0174] Active ingredient RTPeak areaHeightConc.UnitG1L4Glycyrrhizin37.115141217831335485847.137mg / L

[0175] Through this, it was confirmed that the glycyrrhizin content of the licorice extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid was significantly higher than that of the control group.

[0176]

[0177] 3-5. Analysis of Nodakenin Content in Angelica Gigas Extract

[0178] The content of nodakenin included in the control group, the ethanol extract of Angelica gigas Nakai prepared in Example 3 above, and the G1L4 group extracted with the G1L4 eutectic solvent was measured.

[0179] The process for measuring the nodakenin content was carried out in the same manner as in Experimental Example 3-1 above, and the HPLC analysis conditions are shown in Table 14 below.

[0180]

[0181] Column 4.6×250 mm, 5 μm, C18 Mobile Phase 0.1% TFA: Acetonitrile (A:B) Flow rate 1 mL / min Analysis time 25 min Wavelength 330 nm Temperature 30 ℃ Injection volume 10 μl

[0182] As a result, as shown in Table 15, Table 16 and Figure 37 below, it was confirmed that the nodakenin content of the G1L4 group of the Angelica gigas Nakai extract using the natural hydrogen bond donor solvent of the present invention was significantly increased compared to the control group, which was the ethanol extract group.

[0183] Specifically, in the HPLC spectrum, at a retention time (RT) of 3.5 minutes corresponding to Angelica gigas Nakai, the peak area of ​​the G1L4 group increased approximately 1.6 times compared to the control group.

[0184] Active ingredient R T Peak area Height Conc. Unit Control Nodakenin 3.4766646075731922301.299 mg / L

[0185] Active ingredient RTPeak areaHeightConc.UnitG1L4gunnotakenin3.453106369731510251484.525mg / L

[0186] Through this, it was confirmed that the nodakenin content of the Angelica gigas Nakai extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid was significantly higher than that of the control group.

[0187]

[0188] 3-6. Analysis of gomisin A content in Schisandra chinensis extract

[0189] The content of gomisin A included in the control group, the ethanol extract of Schisandra chinensis prepared in Example 3 above, and the G1L3 group extracted with the G1L3 eutectic solvent was measured.

[0190] The process for measuring the content of gomisin A was carried out in the same manner as in Experimental Example 3-1 above, and the analysis conditions of HPLC are shown in Table 17 below.

[0191] Column 4.6×250 mm, 5 μm, C18 Mobile Phase Distilled water:acetonitrile (A:B) Flow rate 1 mL / min Analysis time 70 min Wavelength 220 nm Temperature 30 ℃ Injection volume 10 ㎕

[0192] As a result, as shown in Table 18, Table 19 and Figure 38 below, it was confirmed that the content of gomisin A in the G1L3 group of the Schisandra chinensis extract using the natural hydrogen bond donor solvent of the present invention was significantly increased compared to the control group, which was the ethanol extract group.

[0193] Specifically, in the HPLC spectrum, at a retention time (RT) of 29.4 minutes corresponding to gomisin A, the peak area of ​​the G1L3 group increased by approximately 1.1 times compared to the control group.

[0194] Active ingredient R T Peak area Height Conc. Unit Control group Gomisin A 29.385 1113 1712 887 892 194.461 mg / L

[0195] Active ingredient RTPeak areaHeightConc.UnitG1L3Gungomisin A29.37712265336907970214.406mg / L

[0196] Through this, it was confirmed that the content of gomisin A in the Schisandra chinensis extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid was significantly higher than that in the control group.

[0197]

[0198] 3-7. Analysis of epicatechin content in green tea extract

[0199] The epicatechin content included in the green tea ethanol extract, which is the control group manufactured in Example 3, and the G1L2 group extracted with the G1L2 eutectic solvent was measured.

[0200] The process for measuring epicatechin content was carried out in the same manner as in Experimental Example 3-1 above, and the HPLC analysis conditions are shown in Table 20 below.

[0201] Column 4.6×250 mm, 5 μm, C18 Mobile Phase 0.1% TFA: Acetonitrile (A:B) Flow rate 1 mL / min Analysis time 30 min Wavelength 235 nm Temperature 30 ℃ Injection volume 10 μl

[0202] As a result, as shown in Table 21, Table 22 and Figure 39 below, it was confirmed that the epicatechin content of the green tea extract G1L2 group using the natural hydrogen bond donor solvent of the present invention was significantly increased compared to the control group, which was the ethanol extract group.

[0203] Specifically, in the HPLC spectrum, at a retention time (RT) of 14 minutes corresponding to epicatechin, the peak area of ​​the G1L2 group increased by approximately 1.2 times compared to the control group.

[0204] Active ingredient RT Peak area Height Conc. Unit Control epicatechin 14.30 8 26 133 14 16 8 28 5 19 5.42 3 mg / L

[0205]

[0206] Active ingredient RTPeak areaHeightConc.UnitControl groupEpicatechin13.9453393675362894251.680mg / L

[0207] Through this, it was confirmed that the epicatechin content of green tea extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid was significantly higher than that of the control group.

[0208]

[0209] 3-8. Analysis of baicalin, baicalein, and wogonin contents in golden extract

[0210] The contents of baicalin, baicalein, and wogonin included in the control group, the golden ethanol extract prepared in Example 3, and the G1L4 group extracted with the G1L4 eutectic solvent were measured.

[0211] The process for measuring the content of baicalin, baicalein, and ogonine was carried out in the same manner as in Experimental Example 3-1 above, and the analysis conditions of HPLC are shown in Table 23 below.

[0212] Column 4.6×250 mm, 5 μm, C18 Mobile Phase 0.1% TFA: Acetonitrile (A:B) Flow rate 1 mL / min Analysis time 30 min Wavelength 275 nm Temperature 30 ℃ Injection volume 10 μl

[0213] As a result, as shown in Table 24, Table 25 and Figure 40 below, it was confirmed that the contents of baicalin, baicalein and ogonine in the G1L4 group of golden extracts using the natural hydrogen bond donor solvent of the present invention were significantly increased compared to the control group, which was the ethanol extract group.

[0214] Specifically, in the HPLC spectrum, at a retention time (RT) of 15.7 minutes corresponding to baicalin, the peak area of ​​the G1L4 group increased by about 3 times or more compared to the control group. In addition, at a retention time of 22 minutes corresponding to baicalein, the peak area of ​​the G1L4 group increased by about 1.4 times or more compared to the control group, and at a retention time of 26 minutes corresponding to ogonine, the peak area of ​​the G1L4 group increased by about 2.4 times or more compared to the control group.

[0215] Active ingredient R T Peak area Height Conc. Unit Control Baicalin 15.70 149 250 147 90 640 155 722 mg / L Baicalein 21.96 337 86 182 537 316 75 436 mg / L Ogonin 25.72 72 24 142 52 89 216 33 949 mg / L

[0216] Active ingredient R T Peak area Height Concentration Unit G 1 L 4 Group Baicalin 15.700 15 15 30 19 17 7 16 24 4 8 1.385 mg / L Baicalein 21.95 8 5 4 13 6 25 7 6 7 10 10 7.567 mg / L Ogonin 25.9 7 6 5 12 4 8 7 3 6 5 5 8 6 0 8 1.647 mg / L

[0217] Through this, it was confirmed that the contents of baicalin, baicalein, and ogonine in the golden extract extracted with a natural hydrogen bond donor solvent mixed with glucose and lactic acid were significantly higher than those in the control group.

[0218]

[0219] In summary, these results indicate that when a plant is extracted with a natural hydrogen bond donor solvent prepared by mixing glucose and lactic acid, the content of physiologically active components increases and the antioxidant effect is excellent, and therefore, the plant extract extracted with the natural hydrogen bond donor solvent of the present invention can be usefully used as an antioxidant composition.

[0220]

[0221] The present invention has been described above, focusing on preferred embodiments and experimental examples. Those skilled in the art will appreciate that the present invention can be implemented in modified forms without departing from its essential characteristics. Therefore, the disclosed embodiments should be considered illustrative rather than limiting. The scope of the present invention is set forth in the claims, not the foregoing description, and all differences within the scope equivalent thereto should be construed as being encompassed by the present invention.

Claims

Natural hydrogen bond donor solvent (NHBDS), a mixture of 1.2 or more hydrogen bond donor (HBD) solvents.

2. In paragraph 1, A natural hydrogen bond donor solvent, characterized in that the hydrogen bond donor solvent is selected from the group consisting of monosaccharides, disaccharides, polysaccharides, amino acids, carboxylic acids, and aldehydes.

3. In paragraph 1, A natural hydrogen bond donor solvent, characterized in that the hydrogen bond donor solvent is mixed at a molar ratio of 1 to 3: 1 to 5.

4. In paragraph 1, A natural hydrogen bond donor solvent, characterized in that the above hydrogen bond donor solvent is a mixture of at least one natural hydrogen bond donor solvent selected from monosaccharides, disaccharides, and polysaccharides; and at least one natural hydrogen bond donor solvent selected from amino acids, carboxylic acids, and aldehydes. A method for producing a natural hydrogen bond donor solvent (NHBDS), comprising the step of mixing 5.2 or more types of hydrogen bond donor (HBD) solvents.

6. In paragraph 5, A method for producing a natural hydrogen bond donor solvent, characterized in that the hydrogen bond donor solvent is mixed at a molar ratio of 1 to 3: 1 to 5. A method for extracting useful components, comprising: a step of extracting useful components from animals, plants, prokaryotes, and eukaryotes using a natural hydrogen bond donor solvent in which 7.2 or more types of hydrogen bond donor solvents are mixed; 8. In paragraph 7, A method for extracting useful components, characterized in that the useful components are at least one selected from the group consisting of phytochemicals, lipids, proteins, pigments, pigments, and fragrances.

9. Step for preparing the plant; A step of preparing a natural hydrogen bond donor solvent (NHBDS) by mixing two or more types of hydrogen bond donor solvents; and A method for increasing the content of physiologically active components in a plant extract, comprising a step of adding the natural hydrogen bond donor solvent to the plant and extracting it.

10. In paragraph 9, A method for increasing the content of physiologically active ingredients in a plant extract, characterized in that the plant is at least one selected from the group consisting of Platycodon grandifloras, Centella asiatica, Panax ginseng, Glycyrrhiza uralensis, Angelica gigas, Schisandra chinensis, Camellia sinensis, and Scutellaria baicalensis.

11. In paragraph 9, A method for increasing the content of physiologically active components in a plant extract, characterized in that the natural hydrogen bond donor solvent is an ultrasonic-treated solvent.

12. In paragraph 11, A method for increasing the content of physiologically active components in a plant extract, characterized in that the above ultrasonic treatment is performed for 20 to 40 minutes.

13. In paragraph 9, A method for increasing the content of physiologically active ingredients in a plant extract, characterized in that the natural hydrogen bond donor solvent is added in an amount of 8 to 12 times the weight of the plant.

14. In paragraph 9, A method for increasing the content of physiologically active components in a plant extract, characterized in that the above extraction is performed at 30 to 70°C for 20 to 40 minutes.

15. In paragraph 9, A method for increasing the content of a physiologically active ingredient in a plant extract, characterized in that the physiologically active ingredient is at least one selected from the group consisting of total phenol, total flavonoid, tannin, platycodin, madecassoside, asiaticoside, ginsenoside, glycyrhizin, nodakenin, gomisin A, epicatechin, baicalin, baicalein, and wogonin.

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