KR20210040899A - Method for preparing Citrus peel extract containing increased content of flavonoid - Google Patents

Abstract

A citrus peel extract manufactured by a manufacturing method of the present invention has an increased content of one or more flavonoids selected from the group consisting of Narirutin, Hesperidin, Nobiletin, Tangeretin, and Sinensetin, an increased content of 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one (DDMP), and increased antioxidant activity or xanthine oxidase inhibitory activity and then has excellent physiological activity. Therefore, the extract can be utilized in various industrial fields such as the development of functional food, food material, and cosmetic material.

Description

Method for preparing Citrus peel extract containing increased content of flavonoid}

The present invention relates to a method for preparing an extract of citrus fruit with an increased content of flavonoids.

Citrus fruits such as mandarin oranges and Hallabong are widely cultivated in the southern regions of Korea and Jeju Island.In the process of making citrus fruits such as beverages and citrus chocolate, processed by-products such as citrus peels and juice gourd are generated more than 50,000 tons per year and are discarded as waste. .

The peel of citrus is called “dermis” and has been traditionally used as a medicinal material. The dermis relieves the lumps of gi and strengthens the function of the spleen to treat abdominal swelling, belching, vomiting, nausea, indigestion, flatulence and drowsy symptoms, and thin stool. It also removes cough and phlegm and is diuretic.

Essential oils contained in citrus fruit skin are known to stimulate digestive organs, promote digestion, expectorm, anti-ulcer, secretion of anti-gastric juice, heart rate, increase blood pressure, anti-allergic, promote bile secretion, inhibit uterine smooth muscle, and have antibacterial effects.

These citrus peels are known to be rich in flavonoids, pectins and oils. Citrus peel contains 1 to 3% of flavonoids based on dry weight, and these flavonoids are known to be more than 60 species.

The main flavonoid compounds in citrus fruits are common flavonoids such as rutin and diosmin, and flavonoids specific to citrus fruits such as hesperidin and naringin, and only citrus-specific flavonoids not reported in vegetables or fruits. Flavonoids such as nobiletin and tangeretin, which are the found polymethoxyflavonoids, exist.

Hesperidin is a flavanone glycoside among the flavonoid pigments present in citrus fruits. It is known to inhibit the formation of lipid peroxide and to reduce cholesterol, as well as anti-oxidative effects such as delaying aging, anti-inflammatory effects, capillary protection and anti-cancer effects.

Narirutin is present in various citrus areas of persimmons, and is known for its antioxidant effect, anti-inflammatory effect, bronchial asthma treatment and anticancer effect, and improvement in alcoholic liver disease.

Naringin is the main component of citrus bitter taste and is contained in summer tangerines and grapefruits. Naringin is known to be effective in liver disease, inhibit platelet aggregation, lower blood pressure, and improve lipid metabolism.

Tangeretin is known to have an effect of preventing metastasis of cancer cells by enhancing adhesion of cancer cells, and is also known to exhibit strong antioxidant and anti-inflammatory activities.

The conventional method for preparing an extract containing nobiletin and tangeretin from citrus fruits has a number of problems for industrial application due to low extraction efficiency, long time or high cost, and high energy consumption. Accordingly, the present inventors aim to provide a method for preparing an extract having a high flavonoid content from citrus fruit peel.

Korean Patent Registration No. 10-0770746

It is an object of the present invention to Narirutin, Hesperidin, Nobiletin, Tangeretin and Sinensetin, comprising the step of roasting citrus peel to a temperature of 140 to 155°C. At least one flavonoid selected from the group consisting of, 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one (2,3-dihydro-3,5-dihydroxy It is to provide a method for preparing a citrus peel extract with increased content of -6-methyl-4(H)-pyran-4-one, DDMP), antioxidant activity, or xanthine oxidase inhibitory activity.

Another object of the present invention is to provide a citrus fruit peel extract prepared by the above manufacturing method.

Another object of the present invention is to provide an antioxidant food composition comprising the citrus fruit extract.

Another object of the present invention is to provide an antioxidant cosmetic composition comprising a citrus fruit extract.

In order to achieve the object of the present invention as described above, Narirutin, hesperidin, nobiletin, including the step of rolling the citrus peel to a temperature of 140 to 155 °C in one embodiment , Tangeretin and sinensetin content of one or more flavonoids selected from the group consisting of, 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4 -One (2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one, DDMP) content, antioxidant activity, or xanthine oxidase inhibitory activity of citrus peel extracts Provides a manufacturing method.

In the present invention, "citrus" refers to the fruit of a plant of luck and citrus (Citrus), Gabos (Sphaerocarpa), autumn scent (Gaeulhyang), potato (Benikoji), light orange (Daidai), grapefruit (Paradisi), Giju mandarin ( Kinokuni), Tangerine/Orange (Sinensis), Dangyuja, Erythrosa, Lime R (Floridana), Limonia, Kanpei, Lemon, Lumia , Limetta, Tangor, Banpeiyu, Murcott, Meyer Lemon, Minihyang, Bergamot, Poonensis, Platymamma, Sarcodactylis, Blood Lime, Blood Orange, Bizzaria, Leiocarpa, Pseudogulgul, Nishinokaori, Depressa, Citrus (Insitorum), Aurantium, Jaffa, Ugli, Ethrog, Sudachi, Oroblanco, Unshiu, King Tangerine, Winter Prince (Winter Prince), Suavissima, Deliciosa, Junos, Iyo, Limettioides, Tamuraana, Jabara, Jigak, Jia Jambhiri, Sunki, Tsunokaori, Setoka, Kiyomi, Nippokoreana, Cara Cara, Hystrix, Calamansi Microcarpa), Webberi, Clementina, Cleopatragul (Reshni), Kinoto (Myrtifolia), Key Lime ( Aurantiifolia), Kinnow, Tangerina, Tangelo, Hassaku, Persian lime (Latifolia), Pyeongyul, Pyriformis, Natsudaidai, Harye orange It may be one or more selected from the group consisting of (Unshiu), Hallabong (Shiranuhi), Hallahyang (Setomi), Tachibana and Goldenhyang (Beni Madonna), and grapefruit.

In the present invention, the "peel" is the peel of a citrus fruit and is also called a dermis. Citrus peel is rich in carotenoids, flavonoids, pectin and terpene.

In the present invention, "extraction" is a method obtained by dissolving only specific components in a substance consisting of various components such as plants or minerals. In the present invention, a group consisting of water, an alcohol solvent having 1 to 4 carbon atoms, and a mixed solvent thereof from citrus fruit peel It means extracting by dissolving a physiologically active substance using one solvent selected from. Specifically, the bioactive substance may be a flavonoid.

In the present invention, “flavonoid” is a kind of secondary metabolite of plants or fungi, and has a structure of 15 carbon skeletons consisting of two phenyl rings (A and B) and a heterocyclic ring (C) as shown in Formula 1.

The flavonoid may be any one flavonoid selected from the group consisting of Naritutin, Hesperidin, Nobiletin, Tangeretin, and Sinensetin.

In the present invention, “Naritutin” is a flavonoid contained in a large amount of citrus fruits and has the structure of Formula 2. Nariroutine is not reported in vegetables or fruits other than citrus fruits, and is known to have antioxidant effects, anti-inflammatory effects, bronchial asthma treatment and anticancer effects, and alcoholic liver disease improvement effects.

In the present invention, "hesperidin" is a kind of flavonoid found in citrus fruits, and is a glycoside of flavonones having the structure of formula (3). Hesperidin has physiological actions such as strengthening capillaries, preventing bleeding, and adjusting blood pressure. Hesperidin has been known since ancient times as a yellow pigment and is used in foods, medicines, and cosmetics.

In the present invention, "Nobiletin" is a flavonoid contained in a large amount of citrus fruits and has the structure of Formula 4. The nobiletin is not reported in vegetables or fruits other than citrus fruits, exerts anticancer action against colon cancer, acts as a therapeutic agent for bronchial asthma, and is known to have a recovery effect against memory damage.

In the present invention, "Tangeretin" is a polymethoxyflavonoid that is mainly concentrated in the peel (skin) of citrus fruits, and has a structure of Formula 5, and is known to have anticancer and antioxidant activities.

In the present invention, "Sinensetin" is a methylated flavone, has the structure of Formula 6, is contained in orange oil, and has anti-inflammatory and antiviral effects.

In the present invention, “DDMP(2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one)” has a structure of Formula 7, and its anti-oxidation and anti-cancer effects are known. have.

In the present invention, the “roasting step” is characterized in that the citrus peel is roasted and cooked without oil at a temperature of 140 to 155°C at which heat is transferred.

The roasting step may be performed for 10 to 60 minutes at a temperature of at least 60 RPM at a temperature in which the delivery temperature of the citrus peel is maintained at 120 to 155°C, and this can be performed for 10 to 60 minutes at a temperature of the citrus peel. It can be different.

More specifically, it may be performed for 10 to 43 minutes at 90 RPM so that the temperature of the citrus peel reaches a temperature of 155°C at a temperature of 250°C.

In the case where the pericarp temperature is less than 140° C. at the roasting temperature, the effect of increasing flavonoids is low, and efficiency decreases, and when it exceeds 160° C., a problem of burning citrus peel may occur.

If the RPM is less than 60 RPM during the abrasion, a problem may occur that the rolling speed takes a long time, and in the case of more than that, the speed can be adjusted within a range in which the skin does not burn.

When the roasting time is less than 10 minutes based on 90 RPM, the temperature transfer rate is low, so when extracting raw materials, the extraction efficiency is low, the color and scent are sweet, and there is a problem of not savory. This black and tan taste can cause problems that are too strong.

In an embodiment of the present invention, in order to confirm the change in the extraction yield, antioxidant activity, total polyphenol content, flavonoid content and DDMP content of the extract of citrus peel according to the roasting temperature, at 250° C., the citrus peel reaches 100 to 155° C. The extract was prepared by roasting with PRM, adding water to the roasted citrus peel, heating to 100°C, and hot water extraction for 1 hour. Then, the extraction yield, antioxidant activity, xanthine oxidase inhibitory activity, total polyphenol content, flavonoid content, and DDMP content of each extract were analyzed.

As a result of the extraction yield analysis, it was confirmed that the extraction yield of citrus peels treated with roasting at 100 to 155°C increased as the roasting temperature increased. Specifically, the extraction yield of the untreated sample was measured as 22.4%, and the extraction yield was measured as 31.0% from the treated citrus peels treated with roasting at 155°C, confirming that the extraction yield increased by 8.6% by the roasting treatment.

As a result of the antioxidant activity analysis, the anti-oxidant activity tended to increase in the sample treated with roasting compared to that of the untreated sample, and the antioxidant activity increased as the roasting temperature increased. Specifically, the extract of citrus peel treated with roasting at 155°C increased DPPH or ABTS radical scavenging activity and xanthine oxidase inhibitory activity.

As a result of analyzing the total polyphenol content, it was confirmed that the higher the roasting temperature, the higher the total polyphenol content. Specifically, the total polyphenol content increased by 10.9% to 12.3% compared to the untreated sample at a roasting temperature of 100 to 120°C, and the total polyphenol content was 43.4 to 74.9% compared to the untreated sample at a roasting temperature of 140 to 155°C. Increased. Among them, the total polyphenol content of the sample roasted at 155°C increased 74.9% compared to the untreated control group, confirming that the polyphenol content was the highest.

Therefore, it was confirmed that the roasting treatment at 140 to 155°C was effective in increasing the total polyphenol content, and it was confirmed that 155°C was an excellent roasting temperature condition for increasing the polyphenol.

As a result of analyzing the content of flavonoids, Narirutin, Hesperidin, Nobiletin, Tangeretin, or Sinensetin, the content of flavonoids increased as the aching temperature increased. Was confirmed.

Specifically, the content of flavonoids Narirutin (Narirutin), hesperidin, nobiletin, and Sinensetin in the roasted citrus peel at 140°C to 155°C is in the range of 100 to 120°C. Was confirmed to be higher than that of the treated sample, and it was confirmed that the contents of Narirutin, Hesperidin, Nobiletin, and Sinensetin were the highest in citrus peels treated with roasting at 155°C.

The content of Tangeretin was measured as 0.09 mg/g in the untreated sample, 0.09 mg/g to 0.10 mg/g at 100 to 120°C, and 0.10 mg/g in the sample treated at 140 to 155°C. To 0.12mg/g, and the highest content was 0.12mg/g at 155°C.

That is, when the abrasion treatment was performed at 140 to 155°C, the content of the citrus peel Narirutin, Hesperidin, Nobiletin, Tangeretin, or Sinensetin was untreated. And it was confirmed that it is superior to the roasted sample at 100 to 120°C, of which it was confirmed that the roasted sample at 155°C had the highest flavonoid content.

As a result of analyzing the change in DDMP content, it was confirmed that the DDMP content increased by the abrasion treatment, and it was confirmed that the DDMP content increased as the temperature of the abrasion treatment increased. Specifically, a small amount of DDMP was found in the initially dried sample, but it was confirmed that it increased significantly at a temperature of 140°C to 155°C. As a result of analyzing the content of DDMP (2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one) according to the roasting temperature, when the temperature reaches 155°C from 0℃ It was confirmed that the content increased as the roasting temperature increased. Specifically, a small amount of 0.20mg/g was detected compared to other heat treatment samples, and 1.83mg/g was produced at 120°C. And 9.83mg/g was produced at 140℃, and the highest content was confirmed as 19.17mg/g at 155℃.

That is, when the roasting treatment was performed at 140 to 155°C, it was confirmed that the DDMP content of the citrus peel was superior to that of the untreated sample and the roasted sample at 100 to 120°C, of which it was confirmed that the roasted sample at 155°C had the highest DDMP content. .

Therefore, the citrus peel extract prepared by roasting at 140 to 155°C, specifically 155°C, is characterized in that the extraction efficiency is improved, and the antioxidant effect, total polyphenol content, flavonoid content, or DDMP content is increased.

In addition, the citrus peel extract prepared by roasting at 140 to 155°C, specifically 155°C is characterized by increased xanthine oxidase inhibitory activity.

In the present invention, “xanthine oxidase” is an enzyme that generates excess oxidation radicals in the process of producing uric acid using xanthine as a substrate. Xanthine oxidase, one of the free radical generation systems in vivo, is a non-specific enzyme involved in the metabolism of purine, pyrimidine pteridine aldehyde, and heterocyclic compounds. It catalyzes the reaction of oxidizing hypoxanthine, a metabolite of purine, to xanthine and xanthine to uric acid.

When uric acid is produced more than normal, or when the kidney's ability to excrete uric acid decreases, excess uric acid remains in the blood and increases blood uric acid levels, leading to hyperuricemia or gout.

The citrus peel extract of the present invention has an increased xanthine oxidase inhibitory activity compared to unsalted citrus peel, thereby reducing uric acid in blood or urine, and thus has excellent preventive or ameliorating effects of gout or hyperuricemia.

In the present invention, "Hyperuricemia" means that the concentration of uric acid in the blood is increased. This occurs when the concentration of monosodium urate in the serum exceeds the limited solubility limit. Plasma uric acid saturation at 37°C is about 7mg/dl. Therefore, when this concentration is exceeded, it becomes physicochemically supersaturated. Serum uric acid concentration is known to be relatively high when it exceeds +2 standard deviation than the average serum uric acid concentration in normal subjects. In most epidemiological studies, the upper limit for men is 7 mg/dl and 6 mg/ for women. For this reason, the actual upper limit level of hyperuricemia is defined as 7.0 mg/dl or more.

In the present invention, “Gout” is a metabolic disease in which crystals of uric acid formed by a long-lasting state in a state of high uric acid in the blood cannot be discharged out of the body, and accumulate in various tissues and cause various symptoms. Increased levels of uric acid in the blood may be caused by excessive uric acid production and improper excretion due to abnormalities in the excretion process through the kidneys. A high uric acid level in the blood does not immediately cause gout, and uric acid crystals are easily formed by the high uric acid level, and if the state of deposit in various tissues persists for 10 to 20 years, gout is caused by the triggering factor. This is mostly common in men in their 40s to 50s, but the age of onset is gradually lowering due to changes in diet and environment, and women who take long-term immunosuppressants after menopause or after organ transplant surgery, or women whose renal function has declined due to long-term use of diuretics. It is more likely to develop from

In the present invention, "improvement" refers to any action that improves or benefits the symptoms of an individual with suspicion of hyperuricemia or gout by using a food composition containing the citrus pericarp extract prepared by the above manufacturing method as an active ingredient.

In the present invention, "prevention" refers to any action that suppresses or delays the onset of hyperuricemia or gout by administration of the composition according to the present invention.

In another embodiment of the present invention, it was confirmed that the content of 5-hydroxymethylfurfural (5-(Hydroxymethyl)furfural, 5-HMF) increased by the abrasion treatment, and the 5-HMF content increased as the abrasion treatment temperature increased. It was confirmed that it increased.

As a result of analyzing the content of 5-HMF according to the roasting temperature, it was confirmed that 5-HMF was not detected at 100°C, and 5-HMF was produced at 120°C or higher at 0.16mg/g at 120°C.

Specifically, it was confirmed that the content of 5-HMF increased as the abrasion temperature increased as it was produced at 2.98 to 16.77mg/g at 140°C to 155°C. In addition, it was confirmed that the content was the highest at 16.77mg/g at 155°C.

That is, when roasting at 140 to 155°C, it was confirmed that the 5-HMF content of the citrus peel was superior to that of the untreated sample and the roasted sample at 100 to 120°C. Of these, the roasted sample at 155°C had the most 5-HMF content. It was confirmed that it was high.

Therefore, the citrus peel extract prepared by the method of the present invention is characterized by an increased content of 5-hydroxymethylfurfural (5-(Hydroxymethyl)furfural, 5-HMF) at the same time.

In the present invention, “5-(Hydroxymethyl)furfural, 5-HMF” has the structure of Chemical Formula 8, and is known to have anti-oxidation and anti-cancer effects.

In another aspect of the present invention, it provides a citrus fruit extract prepared by the above manufacturing method.

Citrus fruit peel extract of the present invention can be prepared by a manufacturing method comprising the step of roasting the citrus peel at a temperature of 140 to 155 ℃, specifically citrus peel to reach a temperature of 140 to 155 ℃ 60 RPM or more It may be prepared by roasting, and more specifically, it may be prepared by roasting for 10 to 43 minutes at 90 RMP at 155°C.

In the above-described examples, it was confirmed that the total polyphenol content, antioxidant activity, and flavonoid content were increased by the roasting treatment of the citrus peel extract, and the total polyphenol content of the citrus peel extract of the present invention was compared with the untreated citrus peel, It is characterized by increased antioxidant activity, DDMP, 5-HMF or flavonoid content.

Therefore, the polyphenol may be a flavonoid, and more specifically, one or more flavonoids selected from the group consisting of Narirutin, Hesperidin, Nobiletin, Tangeretin, and Sinensetin. Can be

As described above, it was confirmed that the content of flavonoids, which are physiologically active substances, and the antioxidant effect in the citrus peel extract increased by the roasting treatment as described above, and the citrus peel extract can be used as a food composition and a cosmetic composition for antioxidant purposes.

In another aspect of the present invention, it provides an antioxidant food composition comprising the citrus fruit extract.

The description of the citrus peel extract in the present invention is as described above.

In the above-described embodiment, it was confirmed that the inhibitory activity of xanthine oxidase involved in uric acid production was increased in the citrus fruit skin extract prepared by the above manufacturing method, and the food composition of the present invention improved or prevented hyperuricemia or gout. It is available for use.

The antioxidant food composition may further include ingredients commonly added during food production, such as proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents.

Examples of the carbohydrate include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

For example, when the food composition of the present invention is prepared as a drink, it may further include citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, headworm extract, jujube extract, licorice extract, and the like.

The food composition may be a health functional food for improving or preventing antioxidant, hyperuricemia, or gout.

The health functional food of the present invention includes beverages (including tea or alcoholic beverages), fruits and processed foods thereof (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat, and processed foods thereof, including the food composition. (E.g. ham, sausage, corned beef, etc.), breads and noodles (e.g. udon, soba, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (e.g. butter, cheese, etc.), It may be edible vegetable oil, margarine, vegetable protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.).

In addition, the health functional food of the present invention may be formulated into tablets, pills, powders, granules, powders, capsules, liquid formulations, and the like. They may be formulated further comprising one or more of carriers, diluents, excipients and additives.

Specific examples of the carrier, excipient, diluent and additive are, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium phosphate, calcium Silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate And it is preferable that at least one selected from the group consisting of mineral oil is used.

Another aspect of the present invention provides an antioxidant cosmetic composition comprising a citrus fruit extract.

The description of the citrus peel extract in the present invention is as described above.

The antioxidant cosmetic composition may further include conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , Oil, powder foundation, emulsion foundation, wax foundation, and may be formulated as a spray, but is not limited thereto. In more detail, it may be prepared in the form of a nourishing cream, astringent lotion, a flexible lotion, lotion, essence, nourishing gel or massage cream.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide are used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, toss, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, and crystallites Sex cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, and the like can be used.

Citrus peel extract prepared by the production method of the present invention increases the flavonoid content of Narirutin, Hesperidin, Nobiletin, Tangeretin and/or Sinensetin, 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one (2,3-dihydro-3,5-dihydroxy-6-methyl-4(H) The content of -pyran-4-one, DDMP) or/and the content of 5-hydroxymethylfurfural (5-(Hydroxymethyl)furfural, 5-HMF) is increased, and the physiological activity such as antioxidant effect is excellent. It can be used in various industrial fields such as the development of materials and cosmetic materials.

1 is a result of analyzing the change in the total polyphenol content of the citrus peel extract according to the roasting temperature.
FIG. 2 is a result of analyzing changes in the contents of five flavonoids (hesperidin, Narirutin, Nobiletin, Tangeretin, and Sinensetin) according to the roasting treatment of citrus peel.
3 shows a 2D contour plot of HPLC chromatography of an extract according to the abrasion treatment of citrus peel.
4 shows 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one (2,3-dihydro-3,5-) according to the roasting temperature of citrus peel. This is the result of analyzing the changes in the content of dihydroxy-6-methyl-4(H)-pyran-4-one, DDMP) and 5-hydroxymethylfurfural (5-(Hydroxymethyl)furfural, 5-HMF).

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and the scope of the present invention is not limited by these examples.

Experiment method

<Example 1> Preparation of sample

① Dry sound treatment

The roasting was performed for 28 minutes by setting the heating temperature of the roaster to 250°C and the rotation speed of 90 rpm so that the temperature delivered to the peel of the citrus (Citrus unshiu) reached 100 to 160°C ( Table 1).

In the case of Comparative Example 4, which was heated at 160°C, a problem of burning of the sample occurred, and analysis was performed except for Comparative Example 4 in the subsequent experiment.

Sample name Abrasion temperature (℃) Comparative Example 1 No treatment Comparative Example 2 100 Comparative Example 3 120 Example 1 140 Example 2 145 Example 3 150 Example 4 155 Comparative Example 4 160

② Sample extraction

10 times the weight of distilled water was added to the roasted citrus peels at each temperature, followed by extraction at 100° C. for 1 hour. Each extraction solution was filtered through a filter paper and then concentrated at 60°C using a vacuum rotary concentrator to prepare a concentrated solution.

The concentrate was frozen at -70°C, and then freeze-dried by setting the oven temperature to 40°C using a freeze dryer, and setting the cooling concentration to -80°C.

③ Preparation of sample

A sample was prepared at a concentration of 100 mg/ml by adding a mixed solvent of DMSO and ethanol (1:1) to the freeze-dried extract.

<Example 2> Activity assay

To confirm the change of antioxidant activity of the extract according to the roasting temperature of citrus peel, ABTS radical scavenging activity, DPPH radical scavenging activity, NO scavenging activity, Xanthine oxidase inhibitory activity and NO production inhibitory activity were analyzed.

① ABTS radical scavenging activity analysis

First, an ABTS solution was prepared by mixing equal amounts of 7 mM ABTS and 2.45 mM ammonium persulfate, and then used after allowing radicals to be generated for 24 hours.

Samples diluted in ethanol at various concentrations were dispensed into a 96-well plate by 100 μl, and the ABTS solution was added in the same amount. After reacting in the dark for 10 minutes, absorbance was measured at a wavelength of 745 nm to analyze ABTS radical scavenging activity.

② DPPH radical scavenging activity analysis

First, 100 µl of a sample diluted with methanol to various concentrations was dispensed into a 96-well plate, and 100 µl of a 0.4 mM DPPH solution was added. Then, after reacting in the dark for 10 minutes, absorbance was measured at a wavelength of 517 nm to analyze DPPH radical scavenging activity.

③ Analysis of NO scavenging activity

1% (w/v) sulfanilamide dissolved in 5% (v/v) phosphoric acid and 0.1% (w/v) naphthylethylenediamine dihydrochloride dissolved in distilled water The mixture was mixed to prepare a grease reagent.

Samples of various concentrations diluted with PBS were dispensed into a 96-well plate by 50 µl, and 50 µl of 10 mM sodium nitro-prusside (SNP) solution was added at a time to react at room temperature for 30 minutes. After the reaction was completed, 100 µl of a Grease reagent was added and reacted for 10 minutes, and then absorbance was measured at a wavelength of 545 nm to analyze NO scavenging activity.

④ Analysis of xanthine oxidase inhibitory activity

In order to analyze the xanthine oxidase inhibitory activity according to the abrasion treatment temperature, superoxidase radical scavenging activity and xanthine oxidase inhibitory activity were analyzed.

○ Analysis of xanthine oxidase superoxide scavenging activity

In 100 µl of 200 mM sodium phosphate buffer (pH 7.5) containing 1 mM xanthine, 2 mM EDTA (pH7.5) and 0.3 mM nitroblue tetrazolium (NBT) at various concentrations, 50 After adding mU/ml xanthine oxidase and reacting for 15 minutes, absorbance was measured at 550 nm wavelength to analyze xanthine oxidase superoxide radical scavenging activity.

○ Xanthine oxidase inhibitory activity

Xanthine oxidase inhibitory activity was determined by adding 50 mU/ml xanthine oxidase to 100 μl of 200 mM sodium phosphate buffer (pH 7.5) containing various concentrations of samples and 1 mM xanthine and 2 mM EDTA. Production was induced, and absorbance was measured at a wavelength of 290 nm to analyze xanthine oxidase inhibitory activity.

⑤ Analysis of NO production inhibitory activity

RAW 264.7 cells were dispensed into a 96-well plate to a final concentration of 1.5×10 ⁵ cells/ml, and cultured in DMEM (Dulbecco’s modified Eagle’s medium) medium containing 10% FBS for 24 hours. Then, each sample was treated in a medium containing 10% FBS, and after 1 hour, LPS (200 ng/ml) was treated and cultured for 24 hours, and the supernatant was recovered.

100 ml of the Griess reagent was mixed with 100 ml of the supernatant and reacted for 10 minutes, and the absorbance was measured at 540 nm to analyze the NO production inhibitory activity.

<Example 3> Analysis of Changes in Components of Citrus Peel According to Roasting Temperature

① Total polyphenol content analysis

For the total polyphenol content, 1.0 ml of distilled water and 0.5 ml of 2N Follin-Ciocalteau's phenol reagent were added to 0.5 ml of the sample solution, and reacted at room temperature for 5 minutes. 2.0 ml of a 7% Na ₂ CO ₃ solution was added thereto, mixed, reacted at room temperature for 90 minutes, and absorbance was measured at 750 nm. As a standard substance, a standard curve for each concentration was drawn up using gallic acid, and the total polyphenol content was calculated.

② Flavonoid component analysis

The change of flavonoid content in citrus peels according to the roasting temperature was analyzed using HPLC. A standard solution and a test solution were prepared as follows, and HPLC was performed under the conditions of Table 2.

○ Preparation of standard solution

Two reducing sugar compounds and 13 flavonoids were used as standard reagents.

As a reducing sugar-producing compound, 5-HMF (5-hydroxymethylfurfural) and DDMP ((2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one) were used.

Flavonoids include Rutin, Eriocitrin, Neoeriocitrin, Narirutin, Rhoifolin, Naringin, Hesperidin, Neohesperidin, and Neohesperidin. Neoponcirin, Poncirin, Nobiletin, Tangeretin, and Sinensetin were used.

The standard reagent was prepared with a mixed solvent of methanol and DMSO (1/1, v/v) to prepare 100 mg/ml stock solution and diluted to prepare 0.25, 0.5, 1.0 and 5.0 mg/ml standard solutions.

○ Preparation of test solution

Add 20 ml of the extraction solvent (DMSO: MeOH = 1:1) to 2 g of citrus peel extract and extract by sonication for 10 minutes, filter this and adjust the volume of the filtered solution with a 20 ml volumetric flask. I got it. The prepared solution was filtered through a 0.45 μm syringe filter and used as a test solution.

Device Waters 2695 Alliance system Detector PDA detector column Waters Sunfire C18 (4.6mm × 250mm, 5mm) Mobile phase
(Mobile phase)

A solvent: methanol (Fisher, HPLC grade)
B solvent: tertiary distilled water

Injection volume 10 μl Temperature 40℃ Runtime 40 minutes

Experiment result

<Experimental Example 1> Extraction efficiency according to the roasting temperature

It was confirmed that the extraction efficiency of citrus peels increased as the roasting temperature increased.At 155℃, the extraction efficiency was 8.6% higher than that of Comparative Example 1 without the roasting treatment, confirming that the extraction efficiency was the best (Table 3). .

Abrasion temperature
(℃) No treatment
(Comparative Example 1) 100
(Comparative Example 2) 120
(Comparative Example 3) 140
(Example 1) 145
(Example 2) 150
(Example 3) 155
(Example 4) Sample weight
(g) 50 50 50 50 50 50 50 Extraction weight
(g) 11.2 11.9 12.4 13.5 14.1 14.9 15.5 Extraction efficiency
(%) 22.4 23.8 24.8 27.0 28.2 29.8 31.0

<Experimental Example 2> Activity change according to the roasting temperature

Citrus peels showed a tendency to increase the antioxidant activity in the roasted samples compared to the untreated samples, and the antioxidant activity increased as the roasting temperature increased. Specifically, it was confirmed that Example 4, which was subjected to abrasion treatment at 155°C, was excellent in DPPH or ABTS radical scavenging activity, so that the antioxidant effect was excellent.

In addition, it was confirmed that Example 4, which was subjected to abrasion treatment at 155°C, had an excellent inhibitory effect on the production of super oxide and uric acid.

Specifically, it was confirmed that Example 4 has superior uric acid production inhibitory effect than allopurinol (ALLO), which is a gout treatment, and superoxide scavenging activity compared to butylated hydroxyanisol (BHA), an antioxidant. As a result of confirming this excellence, it was confirmed that the inhibition of xanthine oxidase was excellent.

Xanthine oxidase inhibition can be used for preventing or improving hyperuricemia or gout caused by uric acid accumulation by inhibiting the production of uric acid in blood or urine.

Therefore, the astringent-treated citrus peel extract can be used to improve or prevent hyperuricemia or gout by reducing uric acid in blood or urine through inhibition of xanthine oxidase.

< Experimental Example 3 > Thinness Temperature dependent Total phenol Content change

1 is a result of analyzing total polyphenolics contents (TPC) according to the roasting temperature. In Figure 1, it was confirmed that the higher the roasting temperature, the higher the total polyphenol content. Specifically, at 100 to 120 ℃ 281.5 to 288.18mg / g increased 27.9 to 31.5mg / g compared to Comparative Example 1, and at 140 ℃ to 155 ℃ 367.9 to 488.8mg / g compared to Comparative Example 1 Total poly It was confirmed that the phenol content increased from 111.3 to 192.2 mg/g.

That is, the total polyphenol content increased by 10.9% to 12.3% of the untreated group at a roasting temperature of 100 to 120°C, and the total polyphenol content increased by 43.4% or more compared to the untreated sample at a roasting temperature of 140°C or higher. Therefore, it was confirmed that roasting treatment at 140°C or higher was effective in increasing the total polyphenol content, and the total polyphenol content of the roasted sample at 155°C increased 74.9% compared to the untreated control group, resulting in the greatest increase in polyphenol content. As a result, it was confirmed that 155°C was an excellent temperature condition for increasing polyphenols.

In addition, when the abrasion treatment was performed at 160° C., citrus peels burned and the experiment could not be performed.

<Experimental Example 4> Changes in the content of major components of flavonoids

Content analysis of 13 flavonoids reported to be present in citrus peel was conducted.

Specifically, Rutin, Eriocitrin, Neoeriocitrin, Narirutin, Rhoifolin, Naringin, Hesperidin, Neohesperidin, 13 kinds of flavonoids such as Neoponcirin, Poncirin, Nobiletin, Tangeretin, and Sinensetin were analyzed, but in this experiment Narirutin, Hesperidin (hesperidin), Nobiletin (Nobiletin), Tangeretin (Tangeretin), sinensetin (Sinensetin) only five kinds of flavonoids were detected.

In addition, it was confirmed that the flavonoid content of Narirutin, Hesperidin, Nobiletin, Tangeretin, and Sinensetin increased as the abrasion treatment temperature increased (Fig. 2). .

The content of Narirutin, which is most frequently contained in citrus peels, increased from 12.43mg/g before roasting to 155℃ after roasting to 15.32mg/g, increasing to 2.89 mg/g. In other words, the content of narirutin increased by about 23% in the sample roasted at 155°C compared to the untreated sample.

Hesperidin, a flavonoid found in the widest range of citrus fruits, increased by 3.34 mg/g from 8.16 mg/g before roasting to 11.50 mg/g after roasting at 155°C. That is, the hesperidin content increased by about 41% in the sample roasted at 155°C compared to the untreated sample.

In addition, it was confirmed that the content of the three polymethoxyflavonoid components also increased during the abrasion treatment. Specifically, nobiletin was measured as 0.31mg/g in the untreated sample, and it was measured as 0.30 to 0.31mg/g in the sample treated at 100 to 120°C, confirming that there was little change in content, and more than 140°C. The content increased from 0.33 to 0.39mg/g during the abrasion treatment at. Specifically, the abrasion-treated sample at 155°C had a nobiletin content of 0.39 mg/g, which was increased by about 26% compared to the untreated group.

The content of Tangeretin was measured as 0.09mg/g in the untreated sample, 0.09 to 0.10mg/g at 100 to 120°C, and 0.10 to 0.12mg/g in the sample treated at 140 to 155°C. It was increased, and it was confirmed that the content was the highest at 155°C, 0.12mg/g.

Sinensetin content was measured to be 0.07 mg/g in the untreated sample and in the sample roasted at 100°C and 120°C, but 0.09 mg/g at 140°C, 0.13 mg/g at 145 to 150°C, and at 155°C. It was confirmed that the content increased to 0.15mg/g and increased at 140℃ or higher, and the content was the highest at 155℃.

That is, it was confirmed that the flavonoid content was significantly increased in the citrus peel extract treated at 140°C to 155°C. Specifically, the content of flavonoids Narirutin, hesperidin, nobiletin, and sinensetin was increased in the citrus peel extract treated at 140°C to 155°C. It was confirmed that the contents of Narirutin, Hesperidin, Nobiletin, and Sinensetin were the highest in the treated citrus peel extract.

<Experimental Example 5> Generation of a new compound by abrasion treatment

Next, it was confirmed that two kinds of compounds were generated by the abrasion treatment (155°C) in the 2D contour plot of the HPLC chromatography of FIG. 3. The two compounds were separated and analyzed using prep-LC. As a result of the analysis, the compound was 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one (2,3-dihydro-3,5-dihydroxy-6- It was confirmed that they were methyl-4(H)-pyran-4-one, DDMP) and 5-hydroxymetalfurfural (5-(Hydroxymethyl)furfural, 5-HMF).

As a result of analyzing the content of DDMP and 5-HMF according to the roasting temperature in FIG. 4, about 0.20 mg/g of DDMP was detected at the untreated temperature, and 9.83 mg/g was produced at 140° C. and 19.17 mg at 155° C. The highest content was shown in /g. The content of DDMP was increased from 120°C to 1.83mg/g compared to the untreated temperature sample, but showed a very low content compared to the conditions between 140°C and 150°C. In the case of 5-HMF, it was not detected in untreated samples and citrus roasted at 100°C, but 0.16mg/g of 5 HMF was produced at 120°C, and 2.98 to 16.77mg/g at 140°C to 155°C, resulting in the roasting temperature. It was confirmed that the content of 5-HMF increased as was increased. In addition, it was confirmed that the content was the highest at 16.77mg/g at 155°C.

Claims (7)

Comprising the step of roasting the citrus peel to a temperature of 140 to 155 ℃,
Content of one or more flavonoids selected from the group consisting of Narirutin, Hesperidin, Nobiletin, Tangeretin and Sinensetin, 2,3-dihydro-3,5 -Dihydroxy-6-methyl-4(H)-pyran-4-one (2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one, DDMP) A method of preparing a citrus fruit extract with increased content, antioxidant activity, or xanthine oxidase inhibitory activity. In claim 1,
The citrus peel extract has an increased content of 5-hydroxymethylfurfural (5-(Hydroxymethyl)furfural, 5-HMF) at the same time,
Method for producing citrus peel extract. In claim 1, the rolling step is performed at a temperature of 155 °C,
Manufacturing method. Citrus peel extract prepared by the manufacturing method of claim 1. An antioxidant food composition comprising the citrus fruit extract of claim 4. The food composition according to claim 5, wherein the composition is used for improving or preventing hyperuricemia or gout. An antioxidant cosmetic composition comprising the citrus peel extract of claim 4.

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