KR20160079361A - Method for producing Hibiscus cannabinus extract with enhanced antioxidant activity and kaempferitrin content - Google Patents

Abstract

The present invention relates to a method of producing a Hibiscus cannabinus extract with enhanced antioxidant activity and kaempferitrin contents, comprising: a step (a) of mixing water and soluplus to manufacture an extraction solvent; and a step (b) of adding the extraction solvent manufactured in the step (a) to dried and crushed leaves of Hibiscus cannabinus, and extracting a hibiscus cannabinus extract. The present invention also relates to the Hibiscus cannabinus extract manufactured thereby, and to processed foods comprising the same.

Description

TECHNICAL FIELD The present invention relates to a method for producing an extract of horseshoe crab having enhanced antioxidative activity and campperitrin content,

The present invention relates to a method for producing an anti-oxidant activity, a method for producing an anti-oxidant activity and a camelite extract, And a processed food containing the wool extract.

Kenaf contains calcium, a large amount of protein, vitamins and iron, and contains a large amount of tannins, saponins, polyphenols and alkaloids, and is also possible to be edible. Compared to milk of the same weight, it contains 4 times calcium, iron and various vitamins. Other studies have reported that kenaf leaf extract was used as hepatocyte protective activity, lipid peroxidation, and hematopoietic agent in the treatment of sore throat disorders, bruises, and bile hyperactivity in India and Africa . The major components of the horses are kaempferol, one of the flavonoids that protects against the oxidation of fat and DNA and has a strong antioxidant effect. In addition, afzelin, kaempferitrin, astragalin, . According to recent studies, the yam extract has been reported to have immunomodulatory, antimicrobial and antioxidant properties.

One way to improve the solubility of poorly soluble drugs is to reduce the particle size of the drug to increase the available surface area. However, the reduction of particle size by pulverization showed that the soluble powders tended to aggregate with each other and the air particles interfered with the dissolution. Thus, studies have been made to dramatically increase the solubility of a drug by mixing a water-soluble polymer carrier having a high solubility in a drug having a particle size capable of increasing solubility to a certain level.

As described above, solid dispersion is defined as a technique capable of changing the poor solubility property of the drug to the soluble solubility by dispersing the poorly soluble drug due to the molecular level interaction with the water-soluble carrier. Currently, water-soluble polymeric carriers used for solid dispersion formation include polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), crospovidone, polyvinylpyrrolidone-polyvinyl acetate Copolymers (PVP-PVA), and cellulose derivatives. The conditions of use of the polymer carrier are non-toxic and do not inhibit the pharmacological effect and structure of the drug, and have no pharmacological effect of the polymer itself.

Soluplus is a graft copolymer and has a molecular weight of 90,000 to 140,000. It has a critical micelle concentration because it shows similar action to surfactant. Solubility is high in water and more than 50% in acetone, more than 54% in methanol (MeOH) and more than 25% in ethanol (EtOH). The graft copolymer is characterized by a hydrophilic group and a hydrophilic group, and the viscosity increases rapidly in an aqueous solution of 30% or more. Soluplus can be used as a hydrophilic polymer dispersant (poly eric soluvilizer) to improve the solubility by dispersing a low-solubility substance in a high-solubility water-soluble polymer.

Korean Patent Publication No. 2011-0092482 discloses a kenaf extract having increased antibacterial activity by far-infrared ray drying and a method for producing the same, Korean Patent No. 0840558 discloses a kenaf extract having anti-inflammatory and immunomodulating activity However, this method is different from the method for producing the wool extract in which the antioxidative activity and the camp peritrin content of the present invention are enhanced.

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and an object of the present invention is to provide an extracting solvent in which an appropriate amount of soluplus is mixed with water in order to maximize the use of a cotton leaf and to produce an extract having improved extraction yield and functionality The present invention has been accomplished on the basis of this finding by providing a wool extract obtained by extracting a wool leaf and having improved functionalities such as antioxidant activity and active ingredients of total wool such as phenol, flavonoid and campperitrin, and a processed food using the wool extract.

In order to solve the above-mentioned problems, the present invention provides a method for producing an extracting solvent, comprising the steps of: (a) mixing water and soluplus to prepare an extraction solvent; And (b) adding the extractant prepared in step (a) to the dried and crushed sheep leaves, and then extracting the leaves. The present invention further provides a method for preparing a wool extract having enhanced antioxidative activity and campperitrin content .

The present invention also provides an antioxidant activity produced by the above method and a wool extract having enhanced camp peritrin content.

The present invention also provides a processed food containing the antioxidant activity and the wool extract having enhanced camp peritrin content.

The present invention also relates to a method for preparing an antimicrobial composition comprising the step of adding an extracting solvent obtained by mixing water and soluplus to dried and crushed sheep leaves and then extracting the antioxidative activity and the kaempferitrin ) ≪ / RTI > content.

The present invention also provides a composition for promoting antioxidative activity and a camp peritrin content of a wool extract containing Soluplus as an active ingredient.

Since the extract of the rabbit produced by the method of the present invention is extracted using water without using an organic solvent, it can be taken more safely, the extraction yield is enhanced, the total phenol content and flavonoid content are improved, Such as Kaempferitrin content and antioxidative activity, which are effective ingredients of the present invention, can be easily used for processed foods, cosmetics, and pharmaceuticals using the above-described wool extract.

FIG. 1 is a graph comparing DPPH radical scavenging activity (%) by concentration of yam extract.
Fig. 2 is a graph comparing total polyphenol contents (mg / g) of yam extract.
3 is a graph comparing the total flavonoid content (mg / g) of yam extract.
Soluplus 15%: Water + Soluplus 15% Mixture Solvent Extract, Soluplus 30%: Water + Soluplus 30% Water: Water Extract, MeOH: Methanol Extract, EtOAc: Ethyl Acetate Extract, Hexane: Mixed solvent extract, Soluplus 45%: water + Solu plus 45% mixed solvent extract.
FIG. 4 shows the results of HPLC analysis of the camp peritrin content of the horse radish extract of FIG. 4 and refers to a standard, methanol extract, water extract, and water + Soluplus 45% mixed solvent extract from the top.

In order to achieve the object of the present invention,

(a) mixing water and soluplus to prepare an extraction solvent; And

(b) a step of adding to the leaves of the dried and crushed sheep the addition of the extraction solvent prepared in the step (a) and then extracting the leaves, wherein the antioxidative activity and the production of the camelite extract .

In the method for producing a wool extract of the present invention, the soluplus in step (a) is a polyvinyl caprylactam-polyvinyl acetate-polyethylene glycol graft copolymer, And preferably has a weight average molecular weight of 90,000 to 140,000 g / mol. SoluPlus improves the solubility of crushed sheep leaves and improves the extraction yield of sheep leaves and extracts the active ingredients more effectively.

In the formula, 1 is 10 to 10,000, preferably 100 to 900. [ Also, m is 20 to 20,000, preferably 150 to 1,500. Also, n is from 30 to 30,000, preferably from 300 to 3,000.

In addition, in the method for producing a wool hemp extract of the present invention, the extraction solvent of step (a) may be prepared by adding 40 to 50% of soluplus to water, Preferably, 45% of soluplus is added to water by weight of water and then mixed. When extracting the sheep leaves using the extraction solvent prepared as described above, not only the extraction yield is increased but also the effective components of the horses such as total polyphenol, flavonoid and campperitrin contents are efficiently extracted and the antioxidant activity And the like.

The method for producing the rabbit extract of the present invention is more specifically

(a) adding 40 to 50% of soluplus to water in water and mixing to prepare an extraction solvent; And

(b) adding the extraction solvent prepared in step (a) to dried and crushed sheep leaves, and then extracting the mixture at 70 to 90 ° C for 1 to 3 hours for 1 to 3 times,

More specifically,

(a) adding 45% of soluplus to water in water and mixing to prepare an extraction solvent; And

(b) adding the extraction solvent prepared in step (a) to dried and crushed sheep leaves, and then extracting twice at 80 ° C for 2 hours.

The present invention also provides an antioxidant activity produced by the above method and a wool extract having enhanced camp peritrin content.

The present invention also provides a processed food containing the antioxidant activity and the wool extract having enhanced camp peritrin content. There is no particular limitation on the kind of the processed food. Examples of the foods to which the wool horse extract can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, rice cakes, ice cream, , A drink, an alcoholic beverage, and a vitamin complex, all of which are processed foods in a conventional sense.

The present invention also relates to a method for preparing an antimicrobial composition comprising the step of adding an extracting solvent obtained by mixing water and soluplus to dried and crushed sheep leaves and then extracting the antioxidative activity and the kaempferitrin ) ≪ / RTI > content.

The present invention also provides a composition for promoting antioxidative activity and a camp peritrin content of a wool extract containing Soluplus as an active ingredient. The antioxidant activity and the camp peritrin content of the rabbit extract can be enhanced by extracting the horseshoe with Solux Plus as the extracting solvent. The SolutoPlus may be prepared by adding 40 to 50% of Soluplus to the weight of water to water.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

1. Experimental material

The leaves of kenaf cultivated in Chuncheon city, Kangwon province in 2014 were harvested and ground, and then the sample was crushed and used. Kaempferitrin used for HPLC analysis was purchased from Sigma Chemical Co. (St. Louis, MO, USA).

2. Production of yam extract

The pulverized samples were mixed with 100 ml of each solvent (water, methanol, ethyl acetate, hexane, water + Soluplus 15%, 30%, 45%) 5 g each and then extracted twice at 80 ° C for 2 hours. The extracts were concentrated in a rotary evaporator (EYELA, N-1000, Tokyo, Japan) and then lyophilized (EYELA, FDU-1200, Tokyo, Japan)

3. DPPH free radical scavenging ability

Each extract was dissolved by concentration using 80% methanol. 1 ml of each extract solution was added to a test tube, and 4 ml of 0.15 mM DPPH solution (dissolved in 80% methanol) was added thereto. The mixture was uniformly mixed and allowed to stand at room temperature for 30 minutes, and absorbance was measured at 517 nm. The DPPH radical scavenging activity (%) was expressed as a percentage (%) in the absorbance difference between the sample added and the non-added sample.

DPPH radical scavenging ability (%) = [(1-A / B) x 100]

A: Absorbance of the sample, B: Absorbance of the control

4. Total polyphenol content measurement

The content of total polyphenol compounds was measured by slightly modifying the Folin-Denis method using the phenomenon that the phenolic material reacted with phosphomolybdate and showed blue color. 0.2 ml of the extract was taken in a test tube and 1.8 ml of distilled water was added thereto, followed by mixing well and left at room temperature for 3 minutes. 0.4 ml of a saturated solution of Na ₂ CO ₃ was added and mixed, and 1.4 ml of distilled water was added to make a total of 4 ml. The mixture was allowed to stand at room temperature for 1 hour and the absorbance of the supernatant was measured at 725 nm. The total polyphenol compounds were determined from the standard curves prepared using gallic acid.

5. Total flavonoid content measurement

Total flavonoid content was determined by adding 0.1 ml of 10% aluminum nitrate, 0.1 ml of 1M aqueous potassium acetate and 4.3 ml of 80% ethanol to 0.5 ml of the extract, reacting at room temperature for 40 minutes, Absorbance was measured. At this time, the content of the total flavonoid compound was determined from a standard curve prepared using quercetin.

6. HPLC analysis conditions

The extracts were dissolved in methanol for HPLC and then analyzed by filtration through a 0.45 μm syringe filter (Toyo Roshi Kaisha, Japan). Shimadzu LC system (CBM-20A, Japan) was used for the HPLC analysis and the columns were Prevail C18, 5 ㎛, 4.6 × 150 mm. The mobile phase was A: water, B: acetonitrile, gradient of 16-50% acetonitrile, and absorbance at 280 nm at a flow rate of 1 ml / min.

Example 1: Result of Yield of Yogurt Extract

The yield of water extract was 42.01% and the extraction efficiency was high in the solvent without addition of solubilizing agent. On the other hand, the extract yielded the highest extract yield of 74.57% in water + SoluPlus 45% extract with solubilizing agent (Table 1).

Extraction yield (%) of yam extract Sample Extraction yield (%) Organic solvent Hexane 7.40 Methanol 34.56 Ethyl acetate 7.74 Water + SOLU PLUS water 42.01 Water + soluplus 15% 40.82 Water + soluplus 30% 51.67 Water + soluplus 45% 74.57

Example 2: Measurement results of DPPH free radical scavenging ability of the wool extract

In the case of DPPH radical scavenging ability, the antioxidant activity of methanol> Soluplus 45%> Soluplus 30%> Ethyl acetate> Soluplus 15%>water> hexane showed the difference of antioxidant power in the order of methanol extract> 2 ㎎ / ㎖, 78.53% , And the antioxidant activity was 68.12% at a concentration of 2 ㎎ / ㎖ of SoluPlus 45% extract. The water + SoluPlus extract was assayed for antioxidant activity in proportion to the addition concentration of SoluPlus (Fig. 1).

Example 3: Total polyphenol content of wool extract

The total polyphenol content was found to be: Soluplus 45%> Soluplus 30%>Methanol> Soluplus 15%>Water> Ethyl acetate> Hexane. In organic solvent, methanol was the highest at 24.01 ㎎ / ㎖ DW, but the highest value was obtained at 45.25% of water plus Soluzu plus 57.25 ㎎ / ㎖ DW. The water + SoluPlus extract was measured for total polyphenol in proportion to the addition concentration of SoluPlus (FIG. 2).

Example 4: Total flavonoid content of wool extract

In the total flavonoid content, methanol was>45%>Soluplus>30%> ethyl acetate>water> Soluplus 15%> hexane in the order of methanol> Soluplus 45%, and methanol with the highest flavonoid value was 9.13 ㎎ / g DW It came out. However, a value similar to that extracted from the methanol solvent by water + SOLUPLUS 45% to 8.73 mg / g DW was measured. Water + Solu Plus extracts were measured for total flavonoids in proportion to the addition concentration of SoluPlus (FIG. 3).

Example 5: Analysis of Kaempferitrin content of wool extract

In the case of HPLC analysis, Kaempferitrin, a major component of sheep, was generally extracted from organic solvents and the content of 57.7 mg / g was determined in the methanol extract. However, when the water extract and the water + SoluPlus 45% extract were compared with each other, the contents of 8.5 ㎎ / g and 17.1 ㎎ / g were higher than those of SoluPlus, respectively (Table 2 and Fig. 4 ).

The content of camp peritrin in the wool extract (mg / g) Sample Campperitrin content Methanol extract 57.7 mg / g Water extract 8.5 mg / g Water + Solu Plus 45% extract 17.1 mg / g

Claims (10)

(a) mixing water and soluplus to prepare an extraction solvent; And
(b) extracting the dried and crushed sheep leaves by adding the extraction solvent prepared in the step (a) and then extracting the leaves. The antioxidant activity and the amount of the camel extract of camel fermented with kaempferitrin ≪ / RTI > The method according to claim 1, wherein the soluplus in step (a) is polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer. Wherein the activity and the camp peritrin content are enhanced. 3. The method according to claim 2, wherein the polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft copolymer is a copolymer of the following formula (1): < EMI ID =
[Chemical Formula 1]

(Wherein l is 10 to 10,000, m is 20 to 20,000, and n is 30 to 30,000). The method according to claim 1, wherein the extraction solvent of step (a) is prepared by adding 40 to 50% of soluplus to water in water, and then mixing the solution. The antioxidant activity and the concentration of camp peritrin are increased ≪ / RTI > The method according to claim 1,
(a) adding 40 to 50% of soluplus to water in water and mixing to prepare an extraction solvent; And
(b) a step of adding the extraction solvent prepared in step (a) to dried and crushed sheep leaves and then extracting the mixture at 70 to 90 ° C for 1 to 3 hours for 1 to 3 times. Wherein the activity and the camp peritrin content are enhanced. An antioxidant activity produced by the method according to any one of claims 1 to 5 and an increased amount of camperitrin. A processed food containing the antioxidant activity of claim 6 and a wool extract having enhanced camp peritrin content. Wherein the extract is prepared by adding an extracting solvent in which water and soluplus are mixed to a dried and crushed sheep leaf, and then extracting the extract. The antioxidant activity and the kaempferitrin content of the rape extract are increased Way. A composition for promoting antioxidative activity and camp peritrin content of a wool extract containing Soluplus as an active ingredient. 10. The antioxidant activity and the antioxidant activity of the rabbit extract according to claim 9, wherein the soluplus is a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer. A composition for promoting campferritin content.

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