KR102364978B1 - Method of processing lonicera japonica thunb to increase anti-oxidant and anti-obesity - Google Patents

Abstract

The present invention relates to a method for processing gold and silver coins, comprising the step of high-pressure treatment of Lonicera japonica Thunb.

Description

Method of processing gold and silver coins to enhance antioxidant and anti-obesity activity

The present invention relates to a method for processing gold and silver coins, comprising the step of high-pressure treatment of Lonicera japonica Thunb.

In the living body, a significant amount of active oxygen is generated during the oxidation process for energy production. When active oxygen is instantaneously generated in large amounts or chronically generated, it reacts strongly with cell components to cause damage to cells and tissues, and continuous cell damage can lead to DNA denaturation, lipid oxidation, protein degradation there is.

In the human body, superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT), glutathione reductase, glutathione-S-transferase ), etc., there are antioxidant enzymes against free radicals, but due to various environmental pollutants, smoking, and stress that increase with industrialization, there is a limit to defending against protein degradation and DNA damage only with the activity of antioxidant enzymes in the human body.

Synthetic substances such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) have been developed as antioxidants, but they can cause various side effects on the human body. Research to find safe antioxidants is required.

On the other hand, as mankind gradually develops into an affluent society, obesity has emerged as one of the serious diseases, and the World Health Organization (WHO) has declared obesity as a disease to be treated.

Obesity is a metabolic disease caused by an imbalance in calorie intake and consumption, and is morphologically caused by an increase in the size (hypertrophy) or an increase in the number (hyperplasia) of fat cells in the body. Obesity is not only the most common nutritional disorder in Western society, but also in recent years in Korea, the importance of treatment and prevention has been greatly emphasized as the frequency of obesity is rapidly increasing due to the improvement of diet and westernization of lifestyle due to economic development. there is.

According to the circumstances as described above, research for enhancing antioxidant and anti-obesity activity is required.

Korean Patent Registration No. 10-1861796

It is an object of the present invention to provide a method for processing gold and silver coins, comprising the step of high-pressure treatment for gold and silver coins ( Lonicera japonica Thunb.).

Another object of the present invention is to provide a gold-silver product manufactured by the above processing method.

Another object of the present invention is to provide an extract of gold and silver coins obtained by extracting the processed gold and silver coins.

Another object of the present invention is to provide a food composition comprising the gold or silver flower extract or the gold and silver flower extract.

One aspect of the present invention for achieving the above object provides a method of processing gold and silver coins, comprising the step of high-pressure treatment of gold and silver coins ( Lonicera japonica Thunb.).

Specifically, the high pressure treatment may be performed at 300 MPa to 500 MPa.

Also specifically, the high-pressure treatment may be performed for 1 minute to 40 minutes.

In addition, specifically, the step of leaving at 0 °C to 10 °C before the high-pressure treatment may be further included, and the step of leaving at 0 °C to 10 °C may be made for 20 to 30 hours, but is not limited thereto. .

In the present invention, “ Lonicera japonica ” is a flower of L. japonica Thunb. belonging to the genus lonicera of the Caprifoliaceae family, and is known to be effective in diuresis, gastric ulcer, arthritis, suppurative dermatitis, and bronchitis. Therefore, it has been used in the pharmaceutical or cosmetic industry, but has a unique flavor and slight toxicity.

In the present invention, by high-pressure treatment of gold and silver coins, antioxidant activity and anti-obesity activity were enhanced, and it was applied to food.

In one embodiment of the present invention, it was confirmed that the content of phenolic acid, flavonoid, and organic acid exhibiting anti-obesity activity increased when high-pressure treatment was performed on gold and silver coins, and it was confirmed that the antioxidant activity was enhanced throughout the high-pressure time.

The gold and silver coins used in the present invention may be one or more selected from the group consisting of raw materials for gold and silver coins, extracts, pulverized products, dispersions, pulverized solutions, and fermented products, but is not limited thereto, and may be applied in a modified form as necessary. . The 'raw material for gold and silver coins' refers to the whole or a part of gold and silver coins that have been collected or cultivated, and refers to a state before processing, extraction, and pulverization are performed.

Another aspect of the present invention provides a gold-silver product manufactured by the above processing method.

Specifically, the processed gold and silver coins may have enhanced antioxidant or anti-obesity activity, and chlorogenic acid, caffeic acid, 4,5-dicaffeoylquinic acid, 3, 5-dicaffeoyl quinic acid (3,5-dicaffeoylquinic acid), rutin (rutin), quercetin (quercetin), luteolin (luteolin), quinic acid (quinic acid) and one selected from the group consisting of shikimic acid (shikimic acid) It may be characterized in that the content of the above compounds is increased.

In one embodiment of the present invention, when high-pressure treatment is performed on gold and silver coins, chlorogenic acid, caffeic acid, 4,5-dicaffeoylquinic acid, and 3,5-dicaffeoylquinic acid, which are phenolic acids showing antiobesity activity, increase, and the flavonoid rutin , it was confirmed that quercetin and luteolin increased (Table 5). In addition, it was confirmed that antioxidant activity was also enhanced by confirming that DPPH radical scavenging ability, ABTS radical scavenging ability, ferric reducing antioxidant power (FRAP) value and reducing power were increased throughout the high pressure time ( FIGS. 3 to 6 ).

The chlorogenic acid is a substance composed of an ester bond of caffeic acid and quinic acid, and is known to have effects of inhibiting the production of lipid peroxide, inhibiting cholesterol biosynthesis, antioxidant, anti-cancer and anti-obesity. Chlorogenic acid present in plants forms an ester bond with the carboxyl group of chlorogenic acid with carbohydrates and proteins in the cell wall, and the hydroxyl group of chlorogenic acid forms an ether bond with lignin. When these bonds are broken during high-pressure treatment, the bound chlorogenic acid may be released and the content may increase.

In addition, decaffeoyl quinic acid is a substance consisting of ester bonds of two caffeic acids to quinic acid, and the caffeic acid of 4,5-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid present in gold and silver coins is partially dissolved during high pressure treatment. It can be hydrolyzed to chlorogenic acid to increase the content.

The caffeic acid is also called carbolic acid, and is also known as a carcinogen or antioxidant.

The quinic acid is a hydrolysis product of chlorogenic acid, 4,5-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid, and the chlorogenic acid, 4,5-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid and 3,5- Decaffeoyl quinic acid may be hydrolyzed to increase the content of quinic acid.

The shikimic acid is a derivative of quinic acid, and since the quinic acid is hydrolyzed by high-pressure treatment of gold and silver coins to be converted into lactone quinic acid or shikimic acid, the content of the shikimic acid can be increased.

Another aspect of the present invention provides an extract of gold and silver coins obtained by extracting the processed gold and silver coins with hot water or an organic solvent.

The "extract" is obtained by separating the active ingredients contained in the extracting material by contacting the solvent and the extracting material under specific conditions, and the extract may include the active ingredient in the raw material by the extraction process for the gold and silver extract.

The gold and silver extract can be extracted according to a conventional method known in the art for extracting an extract from a natural product, that is, using a conventional solvent under conditions of conventional temperature and pressure. For example, the gold and silver flower extract may be extracted using water, alcohol, ethyl acetate, acetone, nucleic acid, dichloromethane, or a mixed solvent thereof, which may be modified and applied as necessary.

As the extraction method, various methods such as hot water extraction, cold extraction, reflux extraction, and ultrasonic extraction may be used, and hot water extraction may be preferably used, but is not limited thereto.

In addition, even when the gold and silver coins are processed by high-pressure treatment based on the gold and silver extract, a re-extraction process may be performed according to the needs of those skilled in the art.

Since the anti-oxidation or anti-obesity activity of the processed gold and silver flower of the present invention is enhanced through high-pressure treatment, an extract of the gold and silver flower with enhanced antioxidant or anti-obesity activity can be provided by preparing an extract using the same.

Another aspect of the present invention provides a food composition comprising the processed gold and silver coins or an extract of gold and silver coins. Specifically, the food composition may exhibit antioxidant or anti-obesity activity.

In one embodiment of the present invention, it was confirmed that the antioxidant activity of the gold and silver coins was enhanced by high-pressure treatment (FIGS. 3 to 6), and it was confirmed that the contents of phenolic acid and flavonoids, which are known to exhibit anti-obesity activity, were increased ( Table 3 and Table 5).

The food composition may include a food pharmaceutically acceptable carrier.

The food composition of the present invention includes all forms of functional food, nutritional supplement, health food and food additives, etc., food compositions of this type are known in the art. It can be prepared in various forms according to known conventional methods.

In the present invention, when the processed gold and silver flower or the gold and silver flower extract is used as a food additive, it may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prophylactic, health or therapeutic treatment).

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all health foods in the ordinary sense.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , a carbonation agent used in carbonated beverages, and the like. In addition, it may contain the pulp for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination. In addition, the food can be prepared in the form of tablets, granules, powders, capsules, liquid solutions and pills according to known manufacturing methods. In addition, various conventional flavoring agents or natural carbohydrates may be included as additional ingredients.

Also specifically, the food composition may be a health functional food.

The health functional food refers to a food manufactured and processed using raw materials or ingredients useful for the human body according to the Health Functional Food Act No. 6727, and controls nutrients or physiological action for the structure and function of the human body. It refers to ingestion for the purpose of obtaining useful effects for health purposes such as The health functional food of the present invention may contain normal food additives, and the suitability as a food additive is determined according to the general rules and general test methods of food additives approved by the Ministry of Food and Drug Safety, etc., unless otherwise specified. It is judged according to the relevant standards and standards.

When used as a health functional food or health functional beverage containing a processed gold and silver flower or an extract of gold and silver coins produced by the processing method of the present invention, the compound is added as it is or used with other foods or food ingredients, and is used in a conventional method. can be used appropriately. In addition, the mixing amount may be appropriately determined depending on the intended use thereof (prevention, health or improvement, therapeutic treatment).

The high-pressure treatment method of the present invention can enhance the antioxidant or anti-obesity activity of gold and silver coins, and the processed gold and silver coins and extracts of gold and silver coins prepared thereby can be utilized as antioxidant or anti-obesity food compositions.

It should be understood that the effects of the present invention are not limited to the above-described effects, and include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a result of analyzing the phenolic acid and flavonoid content of Examples and control samples of the present invention through RP-HPLC, specifically, (1) is chlorogenic acid, (2) is caffeic acid, (3) is 4, 5-dicaffeoylquinic acid, (4) is 3,5-dicaffeoylquinic acid, (5) is rutin, (6) is quercetin, and (7) is luteolin.
2 is a result of analyzing the organic acid content of Examples and Controls of the present invention through HPLC.
3 is a result of measuring the DPPH radical scavenging ability of the Example and the control sample of the present invention.
4 is a result of measuring the ABTS radical scavenging ability of the Example and the control sample of the present invention.
5 is a result of measuring ferric reducing antioxidant power (FRAP) of the examples and control samples of the present invention.
6 is a result of measuring the reducing power of the Example and the control sample of the present invention.

Hereinafter, the present invention will be described in detail by way of Examples. However, the following examples only illustrate the present invention, and the present invention is not limited by the following examples.

Analysis of the general composition of gold and silver coins

For general components of gold and silver coins, moisture, crude flour, crude fat, and crude protein contents were analyzed according to the AOAC method, and carbohydrates were calculated by subtracting these contents from 100.

As shown in Table 1 below, the moisture content of gold and silver coins was 6.06%, the crude flour content was 6.20%, the crude fat content was 3.77%, the protein content was 10.35%, and the carbohydrate content was 73.62%.

specimen Ingredients (%) moisture views george crude protein carbohydrate gold coin 6.06 ± 0.15 6.20 ± 0.10 3.77 ± 0.16 10.35 ± 0.13 73.62 ± 0.11

Preparation Example 1. High pressure treatment of gold and silver coins

After standing at 4 °C for 24 hours, 12 g of gold and silver coins were vacuum-packed using a vacuum sealer (SW-600, MAgic Seal, Gyeonggi-do, South Korea), followed by an isostatic pressing machine (CIP22260, Avure Technologies Inc., Lewis). center, OH, USA) at 400 MPa for different pressure times and high pressure treatment.

Specifically, high pressure treatment was carried out for 1, 5, 10, 20 and 40 minutes, respectively, in order, Example 1 (1 minute high pressure treatment), Example 2 (5 minute high pressure treatment), Example 3 (10 minute high pressure treatment), respectively , Example 4 (20 min high pressure treatment) and Example 5 (40 min high pressure treatment).

Experimental Example 1. Analysis of Physicochemical Characteristics of Gold and Silver Coins Treated with High Pressure

1-1. Chromaticity analysis of high-pressure treated gold and silver coins

In order to analyze the physicochemical properties of the high-pressure-treated gold and silver coins, the chromaticity was analyzed, and the high-pressure-treated gold and silver coins were used as a control.

The chromaticity analysis of the gold and silver coins was measured using a Minolta Chroma Meter CR-400 (Konica Minolta, Tokyo, Japan), and the results were L * (brightness), a * (redness), b * (yellowness), ΔE ( total color difference), H ˚ (color angle), and C * (saturation).

ΔE, H˚ and C* were measured by Equations 1 to 3 below (Table 2).

[Formula 1]

ΔE = [( ΔL *) ² + ( Δa *) ² + ( Δb *) ² ] ^0.5

[Equation 2]

H˚ = tan ^-1 ( b* / a* )

[Equation 3]

C* = [( a* ) ² + ( b* ) ² ] ^0.5

division
chromaticity
L *
a *
b *
△ E
H °
C *
control
38.62 ± 1.00 ^a1)
2.60 ± 0.03 ^a
19.47 ± 0.18 ^a
-
82.43 ± 0.06 ^c
19.64 ± 0.18 ^a
Example 1
39.84 ± 0.98 ^a
1.87 ± 0.06 ^b
19.84 ± 0.30 ^a
1.82 ± 0.33 ^a
84.65 ± 0.24 ^a
19.92 ± 0.29 ^a
Example 2
39.37 ± 0.31 ^a
1.98 ± 0.04 ^b
19.78 ± 0.05 ^a
1.21 ± 0.10 ^a
84.33 ± 0.12 ^ab
19.88 ± 0.05 ^a
Example 3
39.20 ± 1.56 ^a
1.93 ± 0.04 ^b
19.33 ± 0.48 ^a
1.59 ± 0.26 ^a
84.34 ± 0.04 ^ab
19.42 ± 0.48 ^a
Example 4
40.17 ± 1.00 ^a
1.92 ± 0.11 ^b
19.93 ± 0.36 ^a
1.53 ± 0.61 ^a
84.54 ± 0.23 ^ab
20.02 ± 0.36 ^a
Example 5
39.23 ± 0.98 ^a
2.00 ± 0.12 ^b
19.59 ± 0.80 ^a
1.44 ± 0.16 ^a
84.21 ± 0.29 ^b
19.69 ± 0.80 ^a
* Values with the same alphabetic character in the same column do not show a significant difference (p<0.05)

Referring to Table 2, the high pressure treatment did not affect the L*, b* and C * values.

The a* value was significantly lower than that of the control group in Examples 1 to 5, and there was no effect due to the high pressure treatment time. It can be seen that the high-pressure treatment significantly increased the chlorophyll content of the gold and silver coins and lowered the redness, thereby reducing the a* value.

The chlorophyll has magnesium at the center, and since it is known that the magnesium is involved in the antioxidant mechanism, the above results suggest that gold and silver coins having an increased chlorophyll content through high-pressure treatment have antioxidant activity.

When the value of ΔE exceeds 5, the human eye can clearly recognize that the color is different from the control. In Examples 1 to 5, ΔE values were all lower than 5.

The above result suggests that the original color could be maintained to some extent even when the gold and silver coins were subjected to high pressure treatment, and the sensory quality characteristics could be preserved.

1-2. Analysis of total phenolic acid and flavonoid content of high-pressure treated gold and silver coins

In order to analyze the physicochemical properties of the high-pressure-treated gold and silver coins, the contents of total phenolic acid and flavonoids were analyzed.

Specifically, the total phenolic acid content of the gold and silver coins was measured by modifying the Folin-Ciocalteau method.

After adding 10 mL of 70% (v/v) methanol to 0.1 g of the example and control samples, the mixture was stirred for 1 minute. Thereafter, after sonication for 30 minutes, centrifugation was performed at 3,500 rpm for 20 minutes. The supernatant was diluted 5 times with distilled water, 0.5 mL of 1 N Folin reagent was added to the 0.5 mL sample solution, and after 3 minutes, 1.5 mL of a 10% (w/v) Na ₂ CO ₃ solution was added. After storage in the dark for 1 hour, absorbance was measured at 725 nm, and gallic acid (GAE) was used as a standard.

In addition, the total flavonoid content of the gold and silver coins was measured by modifying the aluminum chloride colorimetric method.

After adding 10 mL of 70% (v/v) methanol to 0.1 g of the example and control samples, the mixture was stirred for 1 minute. Thereafter, after sonication for 30 minutes, centrifugation was performed at 3,500 rpm for 20 minutes. The supernatant was diluted 5 times with 70% (v/v) methanol, and 0.3 mL of distilled water and 30 uL of 5% (w/v) NaNO ₂ were added to the 0.3 mL sample solution. After 5 minutes, 60 uL of 10% (w/v) AlCl ₃ was added, and after 5 minutes, 200 uL of 1 M NaOH was added. Then, absorbance was measured at 500 nm, and rutin (RE) was used as a standard (Table 3).

division Total phenolic acid content
(ug GAE/100 mg) Total flavonoid content
(ug RE/100 mg) control 2013.08 ± 22.86 ^c3) 4494.79 ± 99.37 ^c Example 1 2158.24 ± 18.83 ^b 4717.01 ± 130.24 ^b Example 2 2203.94 ± 18.05 ^a 5206.60 ± 46.97 ^a Example 3 2151.52 ± 35.97 ^b 5203.13 ± 136.22 ^a Example 4 2142.56 ± 12.49 ^b 4727.43 ± 49.23 ^b Example 5 2149.73 ± 8.39 ^b 4692.71 ± 18.04 ^b * Values with the same alphabetic character in the same column do not show a significant difference (p<0.05)

As shown in Table 3 above, the total phenol and total flavonoid contents of the control group were found to be 2013.08 ug GAE/100mg and 4494.79 ug RE/100mg, respectively. On the other hand, the total phenolic acid and flavonoid contents of Examples 1 to 5 were significantly increased compared to the control group, and in particular, the total phenolic acid and flavonoid contents of Example 2 showed maximum values (2203.94 ug GAE/100 mg and 5206.60 ug RE/100 mg). The above result can be attributed to the cell wall being destroyed by heat treatment, and the phenolic substances bound to the cell wall are converted to a free state.

1-3. Analysis of phenolic acid and flavonoid content using RP-HPLC of high-pressure-treated gold and silver coins

The content of phenolic acid and flavonoids was analyzed in order to analyze the physicochemical properties of the high-pressure treated gold and silver coins.

Specifically, after adding 5 mL of 70% (v/v) methanol to 0.1 g of Example and Control samples, vortexing for 1 minute, sonication for 30 minutes, and centrifugation at 3,500 rpm for 20 minutes. The supernatant was prepared by filtering with a 0.45 um membrane filter (Millipore, Ireland), and the contents of phenolic acid and flavonoids were analyzed using the Aglient 1200 HPLC system (Aglient Technologies, Palo Alto, USA) under the conditions of Table 4 below (Fig. 1 and Table 5). The concentration ranges of each substance for quantitative evaluation of phenolic acid and flavonoids of gold and silver coins were set to 10-500, 0.1-10, 1-100, 10-250, 1-50, 0.1-5, and 0.1-10 ug/mL. .

Phenolic acid is chlorogenic acid, caffeic acid, 4,5-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid, As flavonoids, rutin, quercetin and luteolin were used as standard substances, all of which are known to be anti-obesity-related substances.

column Agilent Eclipse XDB-C ₁₈ column (250 x 4.6 mm, id, 5 um) concentration gradient Phosphoric acid (A: 0.2%, v/v) and methanol (B)
0-30 min: 30-60% B. wavelength 340 nm flow rate 1 mL/min injection volume 10 uL temperature 25 ^o C

division
Phenolic acid content (ug/100 mg)
Flavonoid content (ug/100 mg)
chlorogenic acid
caffeic acid
4,5-decaffeoyl quinic acid
3,5-decafoil quinic acid
Routine
quercetin
luteolin
control
1006.51 ± 37.31 ^c1)
1.92 ± 0.12 ^d
13.38 ± 0.43 ^c
648.77 ± 21.31 ^c
36.07 ± 2.51 ^c
6.49 ± 0.34 ^d
0.09 ± 0.02 ^c
Example 1
1145.11 ± 46.70 ^b
2.12 ± 0.15 ^cd
15.50 ± 0.77 ^b
765.47 ± 39.18 ^b
39.51 ± 0.80 ^b
7.86 ± 0.32 ^b
0.11 ± 0.01 ^b
Example 2
1244.69 ± 15.84 ^a
2.60 ± 0.07 ^a
17.20 ± 0.27 ^a
833.76 ± 14.50 ^a
43.60 ± 0.22 ^a
7.85 ± 0.48 ^b
0.15 ± 0.01 ^a
Example 3
1241.15 ± 29.47 ^a
2.50 ± 0.10 ^ab
17.73 ± 0.79 ^a
837.64 ± 34.57 ^a
42.32 ± 0.06 ^ab
8.35 ± 0.20 ^a
0.13 ± 0.02 ^ab
Example 4
1229.58 ± 44.74 ^a
2.30 ± 0.15 ^bc
17.39 ± 0.73 ^a
825.50 ± 28.85 ^a
42.89 ± 3.45 ^a
7.92 ± 0.10 ^b
0.11 ± 0.01 ^b
Example 5
1159.08 ± 0.31 ^b
2.10 ± 0.11 ^cd
16.14 ± 0.03 ^b
762.78 ± 3.32 ^b
40.66 ± 0.65 ^ab
6.93 ± 0.14 ^c
0.12 ± 0.01 ^b
* Values with the same alphabetic character in the same column do not show a significant difference (p<0.05)

As shown in Figure 1, gold and silver coins contain phenolic acids (chlorogenic acid, caffeic acid, 4,5-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid) and flavonoids (rutin, quercetin and luteolin). Confirmed. Chlorogenic acid is one of the main substances of gold and silver coins, and is composed of an ester bond between caffeic acid and quinic acid.

As shown in Table 5, the phenolic acid and flavonoid contents of the gold coins were significantly increased throughout the high pressure treatment time (Example 2).

In particular, the contents of chlorogenic acid, caffeic acid, 4,5-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, rutin, and luteolin showed the maximum content at the time of high pressure treatment for 5 minutes, and the maximum amount of quercetin at the time of high pressure treatment for 10 minutes. content was shown.

The above results showed that the content of chlorogenic acid, caffeic acid, 4,5-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, rutin, quercetin and luteolin, which are related to anti-obesity, was significantly reduced by controlling the high-pressure treatment time of gold and silver coins. It suggests that it can be used as a remarkably excellent composition for suppressing obesity.

1-4. Analysis of organic acid content using HPLC of high-pressure treated gold and silver coins

In order to analyze the physicochemical properties of the high-pressure treated gold and silver coins, the content of organic acids was analyzed.

Specifically, 5 mL of distilled water was added to 0.1 g of Example and Control samples, followed by vortexing for 1 minute, sonication for 30 minutes, and centrifugation at 3,500 rpm for 20 minutes. The supernatant was prepared by filtering with a 0.45 um membrane filter (Millipore, Ireland). The content of organic acids was analyzed under the conditions shown in Table 6 below using the Aglient 1200 HPLC system (Aglient Technologies, Palo Alto, USA) (FIG. 2).

Quinic acid, malic acid, shikimic acid and citric acid were used as standard materials for organic acids.

column Zorbax SB-Aq column (150 x 4.6 mm, i.d., 5 um) isocratic 200 mM aqueous phosphate buffer (pH 2.0)/acetonitrile (99:1, v/v) wavelength 214 nm, ref 360 nm flow rate 0.6 mL/min injection volume 10 uL temperature 25 ^o C

Referring to FIG. 2 , the contents of quinic acid, malic acid, shikimic acid and citric acid were not affected by the high pressure treatment, which could be attributed to the fact that the hydrolysis of chlorogenic acid did not occur much during the high pressure treatment and the organic acid content did not increase. there is.

Experimental Example 2. Evaluation of antioxidant activity of high-pressure treated gold and silver coins

The antioxidant activity of the high-pressure treated gold and silver coins was evaluated.

Specifically, 10 mL of 70% (v/v) methanol was added to 0.1 g of Examples 1 to 5 and the control (gold and silver coin not subjected to high pressure treatment) samples. After vortexing for 1 minute, sonicated for 30 minutes, and centrifuged at 3,500 rpm for 20 minutes. The supernatant was diluted 10-fold with 70% (v/v) methanol and then used to evaluate each antioxidant activity.

2-1. Measurement of DPPH radical scavenging ability

The DPPH radical scavenging ability of high-pressure-treated gold and silver coins was measured by modifying the Blois method (Blois MS 1958).

Specifically, after adding 0.6 mL of 200 mM DPPH to 0.2 mL of Examples 1 to 5 and the control sample, it was stored in a dark room for 15 minutes. Then, absorbance was measured at 517 nm, and compared and calculated as ascorbic acid equivalent antioxidant capacity (AEAC) for ascorbic acid, an antioxidant ( FIG. 3 ).

Referring to FIG. 3 , in the case of Examples 1 to 5 treated with high pressure, the DPPH radical scavenging ability was significantly superior to that of the control group, and in particular, it was confirmed that the DPPH radical scavenging ability was the maximum in Example 1.

2-2. ABTS radical scavenging activity measurement

ABTS radical scavenging activity of gold and silver coins treated with high pressure was published in Indian Journal of Geo-Marine Sciences Vol. 42(5), September 2013, pp. 556-564 The method of "Comparison of DPPH and ABTS assays for determining antioxidant potential of water and methanol extracts of Spirulina platensis " was modified and measured.

Specifically, to prepare ABTS ⁺ solution, 7 mM ABTS reagent and 2.45 mM potassium persulfate were mixed at a ratio of 1:1 (v/v), and then left in a dark room for 12 to 16 hours. Then, the prepared ABTS ⁺ reagent was diluted with methanol to adjust the absorbance value at 734 nm to 0.700±0.005.

After adding ABTS ⁺ 0.9 mL to 0.1 mL of Examples 1 to 5 and the control sample, it was stored in a dark room for 6 minutes. Absorbance was measured at 734 nm and compared and calculated as Trolox equivalent antioxidant capacity (TEAC) for the antioxidant Trolox (FIG. 4).

Referring to FIG. 4 , Examples 1 to 5 showed remarkably superior ABTS radical scavenging ability compared to the control group, and in particular, Example 1 showed the greatest ABTS radical scavenging ability.

2-3. FRAP (Ferric reducing antioxidant power) measurement

Ferric reducing antioxidant power (FRAP) values of Examples 1 to 5 and the control group are ANALYTICAL BIOCHEMISTRY 239, 70-76 (1996) 「The Ferric Reducing Ability of Plasma (FRAP) as a Measure of ''Antioxidant Power'': The FRAP Assay" was modified and measured.

Specifically, the FRAP reagent is 10 mM 2,4,6,-tripyridyl-s-triazine (TPTZ) and 20 mM FeCl ₃ dissolved in 300 mM sodium acetate buffer (pH 3.6) and 40 mM hydrochloric acid 10: It was prepared by mixing 1:1 (v/v/v) and heating at 37°C.

After adding 2.85 mL of the FRAP reagent prepared above to 0.15 mL of Examples 1 to 5 and the control sample, it was stored in a dark room at 37°C for 15 minutes. Absorbance was measured at 593 nm, and FeSO ₄ ·7H ₂ O (0 to 1 mM) was used as a standard material (FIG. 5).

Referring to FIG. 5 , in the case of Examples 1 to 5, the FRAP value was significantly increased compared to the control group, and in particular, the FRAP value of Example 1 was the maximum.

2-4. reducing power measurement

The reducing power of Examples 1 to 5 and the control was determined by Sanjukta et al. (2015), Journal of Functional Foods , 14 , 650-658 "Enhancement of antioxidant properties of two soybean varieties of Sikkim Himalayan region by proteolytic Bacillus subtilis fermentation" was modified and measured.

Specifically, 0.9 mL of 200 mM phosphate buffer (pH 6.6) and 0.9 mL of 1% (w/v) potassium ferric cyanide were added to 0.1 mL of the samples of Examples 1 to 5 and the control group. Then, 0.9 mL of 10% (w/v) trichloroacetic acid was added and centrifuged at 3,500 rpm for 20 minutes. The supernatant (0.9 mL) was mixed with 0.9 mL of distilled water and 0.9 mL of 0.1% (w/v) FeCl ₃ , and absorbance was measured at 700 nm. Ascorbic acid (0-100 ug/mL) was used as a standard material (FIG. 6).

Referring to FIG. 6 , Examples 1 to 5 showed excellent reducing power compared to the control group, and in particular, Example 1 showed the maximum reducing power.

The above results confirm that the antioxidant activity of the gold and silver coins is enhanced by processing the gold and silver coins using the high-pressure treatment method of the present invention.

The description of the present invention described above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and likewise components described as distributed may be implemented in a combined form.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present invention.

Claims (11)

High pressure treatment on gold and silver coins ( Lonicera japonica Thunb.);
As a method of processing gold and silver coins, the high pressure treatment is performed at 300 MPa to 500 MPa for 1 minute to 40 minutes,
Further comprising the step of leaving at 0 ℃ to 10 ℃ before the high pressure treatment,
The step of leaving at 0 ℃ to 10 ℃ will be made for 20 to 30 hours, the processing method of gold and silver coins. delete delete delete The method of claim 1,
The method for processing gold and silver coins, wherein the gold and silver coins are at least one selected from the group consisting of raw materials for gold and silver coins, extracts, pulverized products, dispersions, pulverized solutions, and fermented products. A processed product of gold and silver coins manufactured by the processing method of claim 1. The processed gold and silver coins according to claim 6, wherein the processed gold and silver coins have enhanced antioxidant activity. 7. The method according to claim 6, wherein the processed gold and silver coins are chlorogenic acid, caffeic acid, 4,5-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid (3) ,5-dicaffeoylquinic acid), rutin (rutin), quercetin (quercetin), luteolin (luteolin), quinic acid (quinic acid) and that the content of one or more compounds selected from the group consisting of shikimic acid (shikimic acid) is increased Characterized by a gold and silver coin work. An extract of gold and silver coins obtained by extracting the processed product of claim 6 with hot water or an organic solvent. A food composition comprising the processed product of claim 6 or the extract of claim 9 . The food composition according to claim 10, wherein the food composition exhibits antioxidant activity.

Images