KR102450185B1 - Method for extracting active ingredients from Ulmi Cortex - Google Patents

Abstract

The present invention provides a method for extracting an active component of Ulmus pumila comprising the following steps: preparing Ulmus pumila; extracting a solution containing an active component by mixing the prepared Ulmus pumila with an organic solvent; and concentrating and freeze-drying the extracted solution under reduced pressure to manufacture an Ulmus pumila extract.

Description

Method for extracting active ingredients from milk white skin {Method for extracting active ingredients from Ulmi Cortex}

The present invention relates to a method for extracting an active ingredient from milky white skin.

Ulmi Cortex belonging to the family Ulmaceae is the bark or root bark of elm, and has been reported to have excellent effects in water bathing, expectoration, anticancer, anti-corrosive properties, wound healing agents and inflammation. Sitosterol, phytosterol, stimasterol, tannin, starch, polysaccharide, etc. exist, and there have been many studies on the separation and chemical composition of frieselin, epifriendelalol, and taraxerol, which are components that exhibit analgesic action. Attempts have yet to be made.

On the other hand, although it has antioxidant enzymes that protect against free radicals in the human body, such as superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT), glutathione reductase, and glutathione-S-transferase, various environmental pollution that is increasing with industrialization Due to substances, smoking, and stress, the role of the antioxidant system in the human body alone exceeds the defense system, causing protein degradation and DNA damage.

Therefore, synthetic substances such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) have been developed as antioxidants. is becoming

On the other hand, inflammation is a defense mechanism that prevents damage to human tissue due to infection and is accompanied by symptoms such as development, fever, and pain. If this inflammatory response is repeated for a long time, it becomes a factor that can cause various chronic diseases such as neurodegenerative diseases and cancer.

Macrophages are involved in and regulate the production of inflammatory mediators such as nitric oxide (NO), prostaglandin (PG), and pro-inflammatory cytokines in the immune response in the body. These inflammatory mediators induce an inflammatory response, and when the host's immune response does not respond properly, it causes inflammatory diseases.

On the other hand, in order to utilize a material with excellent physiological activity as an industrial material, research and review on the optimal manufacturing process considering the characteristics of the material is required separately from the search for physiological activity. Biological materials have limitations during the production process, so it is necessary to produce a large amount of functional materials with a small space and low cost, and it is necessary to develop an extraction process that can produce as much useful material as possible from the same amount of biological material at a low cost. .

Patent Document 1: Republic of Korea Patent Publication 10-2018-0106104

Accordingly, the present inventors have tried to provide a method for effectively extracting the main ingredients from the milkweed, and as a result, the extraction process, process temperature, extraction solvent, and additives were optimized to determine the extraction amount of polyphenols and flavonoids, which are the main ingredients and active ingredients. It has been found that improvements can be made and the present invention has been completed.

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In order to achieve the above object, the present invention

Preparing milk white blood;

Extracting a solution containing an active ingredient by mixing the prepared milkweed with an organic solvent; and

Concentrating the extracted solution under reduced pressure and freeze-drying to prepare a milkweed extract; provides a method for extracting milkweed active ingredients comprising.

In addition, extracting the solution containing the active ingredient is performed by mixing the prepared milkweed with an organic solvent and a eutectic solvent, and the organic solvent is a mixture of acetone and chloroform in a volume ratio of 1:1, and the eutectic The solvent is a mixture of choline chloride as a salt and ethylene glycol as a hydrogen bond donor in a molar ratio of 1:4,

The extraction is characterized in that the ultrasonic process is performed for 24 hours at a temperature of 30 ℃.

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Milkweed extract according to the present invention shows a very high content of major components with a total polyphenol content of 167.54 mg/g and a total flavonoid content of 25.31 mg/g.

the present invention

Preparing milk white blood;

Extracting a solution containing an active ingredient by mixing the prepared milkweed with an organic solvent; and

Concentrating the extracted solution under reduced pressure and freeze-drying to prepare a milkweed extract; provides a method for extracting milkweed active ingredients comprising.

Hereinafter, the method for extracting the active ingredient of milk white skin according to the present invention will be described in detail.

First, the method for extracting milky white skin active ingredient according to the present invention includes the step of preparing the milky white skin.

In the above step, after cutting the milkweed to an appropriate size, freeze-drying, and grinding with a blender can be prepared in powder form.

Next, the method for extracting the active ingredient of milkweed according to the present invention comprises the step of extracting a solution containing the active ingredient by mixing the prepared milkweed with an organic solvent.

In the above step, to extract a solution containing the active ingredient by mixing the prepared milkweed with an organic solvent, increasing the active ingredient and/or active ingredient extracted from the extract of the milkweed should be performed, and an organic solvent (or extraction solvent) The type and content of additives mixed therewith, extraction process, and process temperature can be the main conditions.

As the organic solvent (or extraction solvent), methanol, ethanol, acetone, chloroform, ethylene chloride, ethyl acetate, n-hexane, dichloromethane, etc. may be used, and preferably acetone and chloroform are mixed in a volume ratio of 1:1. that can be used

The extraction may be performed by a dipping process, an ultrasonic process, a reflux process, etc., and preferably, the extraction may be performed through an ultrasonic process.

The extraction may be performed for 24 hours at a temperature range of 20-40 °C, preferably at a temperature of 30 °C for 24 hours.

For the extraction, a eutectic solvent may be used as an additive, and the eutectic solvent includes a salt and a hydrogen bond donor. The salt in the eutectic solvent is choline chloride (ChCl), tetramethylammonium chloride (Tetramethylammonium chloride, Me4NCl), methyl triphenyl phosphonium bromide (Methyl triphenyl phosphonium bromide, Me (Ph) 3 PBr) and the like can be used, Preferably, choline chloride may be used. In the eutectic solvent, as the hydrogen bond donor, ethylene glycol, 1,2-butanediol, phenol, urea, etc. may be used, and preferably ethylene glycol may be used. can Preferably, choline chloride and ethylene glycol may be mixed in a molar ratio of 1:4 for extraction of the milkweed.

The content of the eutectic solvent may be 0.01-1.0 g per 1 ml of the organic solvent, and in a preferred example, the content of the eutectic solvent may be 0.6 g per 1 ml of the organic solvent.

Next, the method for extracting milky white skin active ingredient according to the present invention includes the step of concentrating the extracted solution under reduced pressure and freeze-drying to prepare a milky white skin extract.

Finally, the extracted solution is concentrated and freeze-dried under reduced pressure to prepare a milkweed extract, and the extract may be filtered with a filter paper and then concentrated under reduced pressure and freeze-dried at 55°C with a concentrator.

The milkweed extract prepared through this is characterized in that it contains a total polyphenol content of 167.54 mg/g and a total flavonoid content of 25.31 mg/g.

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Hereinafter, the present invention will be described in more detail by way of the following examples.

However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited by the examples and experimental examples.

<Example 1> Preparation of milk white skin extract-1

After preparing the milk white skin and freeze-drying, it was ground with a blender to prepare milk white skin powder.

0.05 g of the prepared milk powder was added to 5 ml of 70% ethanol as an organic solvent.

Then, dipping, ultrasonic processing (ultra sonic) and reflux (reflux) processes are performed to extract for 24 hours at a temperature of 20-40 ° C., and then filter with filter paper (Watman No. 2) and concentrator (EYELA N -3000, Japan) was concentrated under reduced pressure at 55 °C and freeze-dried to obtain an extract. The conditions for extracting milk white skin carried out in Example 1 are shown in Table 1 below.

The total polyphenol content and total flavonoid content of the prepared milkweed extract were analyzed. Specifically, the total polyphenol content was measured by applying the Folin-Denis method, and the total flavonoid content was measured by applying the Nieva Moreno method. is shown in Table 2 below.

Example 1 extraction solvent extraction process Process temperature (℃) Example 1-1 ethanol dipping 20 Example 1-2 ultrasonic wave 20 Examples 1-3 reflux 20 Examples 1-4 ultrasonic wave 10 Examples 1-5 ultrasonic wave 30 Examples 1-6 ultrasonic wave 40

Example 1 Total polyphenol content
(mg/g) Total flavonoid content
(mg/g) Example 1-1 88.52 14.28 Example 1-2 90.70 14.89 Examples 1-3 86.21 10.47 Examples 1-4 87.22 12.56 Examples 1-5 91.35 15.90 Examples 1-6 90.86 14.28

As shown in Table 1 and Table 2, when the ultrasonic process is applied as the extraction process and 30° C. is applied as the process temperature, the total polyphenol content is 91.35 mg/g and the total flavonoid content is 15.90 mg/g under the ethanol extraction solvent. It can be seen that the extraction efficiency is shown.

<Example 2> Preparation of milkweed extract-2

After preparing the milk white skin and freeze-drying, it was ground with a blender to prepare milk white skin powder.

0.05 g of the prepared milk powder was added to 5 ml of an organic solvent containing at least one of ethanol, methanol, acetone, chloroform, ethylene chloride, n-hexane, and dichloromethane.

Then, after performing an ultrasonic process to extract at a temperature of 30 ℃ for 24 hours, filtered with filter paper (Watman No. 2), concentrated under reduced pressure at 55 ℃ with a concentrator (EYELA N-3000, Japan) and freeze-dried to extract the extract got it The conditions for extracting milk white skin performed in Example 2 are shown in Table 3 below.

The total polyphenol content and total flavonoid content of the prepared milkweed extract were analyzed. Specifically, the total polyphenol content was measured by applying the Folin-Denis method, and the total flavonoid content was measured by applying the Nieva Moreno method. is shown in Table 4 below.

Example 2 extraction solvent extraction process Process temperature (℃) *Examples 1-5 ethanol ultrasonic wave 30 Example 2-1 methanol Example 2-2 acetone Example 2-3 chloroform Example 2-4 ethylene chloride Example 2-5 n-hexane Example 2-6 dichloromethane **Example 2-7 Acetone/Chloroform

* Examples 1-5 are taken from Example 1.

** Example 2-7 is a mixture of acetone and chloroform in a 1:1 volume ratio

Example 2 Total polyphenol content
(mg/g) Total flavonoid content
(mg/g) Examples 1-5 91.35 15.90 Example 2-1 82.18 14.22 Example 2-2 92.37 16.35 Example 2-3 91.90 16.05 Example 2-4 78.41 10.48 Example 2-5 83.48 11.57 Example 2-6 82.59 10.95 Example 2-7 93.75 16.80

As shown in Tables 3 and 4, it was confirmed that excellent extraction efficiency was exhibited when ethanol, acetone and chloroform were applied as extraction solvents. Furthermore, when a solvent mixed with acetone and chloroform in a volume ratio of 1:1 is applied as the extraction solvent, the extraction amount is significantly increased by showing the highest extraction amount, 93.75 mg/g of total polyphenol content, and 16.80 mg/g of total flavonoid content. could confirm that

<Example 3> Preparation of milk white skin extract-3

After preparing the milk white skin and freeze-drying it, it was ground with a blender to prepare milk white skin powder.

0.05 g of the prepared milk powder and 0.01-1.00 g of the eutectic solvent were added to 5 ml of a solvent in which acetone and chloroform were mixed in a volume ratio of 1:1.

Then, after performing an ultrasonic process to extract at a temperature of 30 ℃ for 24 hours, filtered with filter paper (Watman No. 2), concentrated under reduced pressure at 55 ℃ with a concentrator (EYELA N-3000, Japan) and freeze-dried to extract the extract got it The conditions for the extraction of milk white skin carried out in Example 3 are shown in Table 5 below.

The total polyphenol content and total flavonoid content of the prepared milkweed extract were analyzed. Specifically, the total polyphenol content was measured by applying the Folin-Denis method, and the total flavonoid content was measured by applying the Nieva Moreno method. is shown in Table 6 below.

Example 3 eutectic solvent Salt: hydrogen bond donor molar ratio Amount of eutectic solvent added (g)
salt hydrogen bond donor Example 3-1 Choline Chloride ethylene glycol 1:4 0.01 Example 3-2 tetramethylammonium chloride ethylene glycol 1:4 0.01 Example 3-3 Methyl triphenyl phosphonium bromide ethylene glycol 1:4 0.01 Example 3-4 Choline Chloride 1,2-Butanediol 1:2 0.01 Example 3-5 Choline Chloride urea 1:2 0.01 Example 3-6 Choline Chloride ethylene glycol 1:4 0.05 Example 3-7 0.1 Example 3-8 0.2 Example 3-9 0.3 Example 3-10 0.4 Example 3-11 0.6 Example 3-12 1.0

Example 3 Total polyphenol content
(mg/g) Total flavonoid content
(mg/g) Example 3-1 95.80 18.25 Example 3-2 93.85 16.95 Example 3-3 92.54 16.57 Example 3-4 94.69 17.20 Example 3-5 93.95 17.02 Example 3-6 100.27 19.87 Example 3-7 110.75 20.43 Example 3-8 124.05 20.98 Example 3-9 136.60 21.30 Example 3-10 146.23 22.47 Example 3-11 167.54 25.31 Example 3-12 152.65 23.01

As shown in Tables 5 and 6, it was confirmed that the extraction efficiency was more excellent when the eutectic solvent was applied. In particular, when 0.6 g of a eutectic solvent in which choline chloride and ethylene glycol are applied in a molar ratio of 1:4 as the eutectic solvent is used, the total polyphenol content is 167.54 mg/g and the total flavonoid content is 25.31 mg/g, indicating that the extraction amount is significantly increased. could

Above, a specific part of the present invention has been described in detail, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Accordingly, it is intended that the substantial scope of the present invention be defined by the appended claims and their equivalents.

Claims (3)

Preparing milk white blood;
The prepared milkweed is mixed with an organic solvent in which acetone and chloroform are mixed in a volume ratio of 1:1, and a eutectic solvent in which choline chloride and ethylene glycol are mixed with a hydrogen bond donor in a molar ratio of 1:4 as a salt, but the content of the eutectic solvent is Extracting a solution containing the active ingredient for 24 hours at a temperature of 30 ℃ by ultrasonication using 0.12 g with respect to 1 ml of the organic solvent; and
Concentrating and freeze-drying the extracted solution under reduced pressure to prepare a milkweed extract, wherein the milkweed extract has a total polyphenol content of 167.54 mg/g and a total flavonoid content of 25.31 mg/g; including; A method for producing a milkweed extract. delete delete