KR19990080214A - Functional drink containing health extract of citrus peel - Google Patents

Abstract

The present invention relates to a functional beverage for health improvement manufactured using the extract of citrus fruit, and more particularly, to a functional beverage for health improvement manufactured using bioflavonoids derived from citrus fruit extract or citrus peel. will be.

Description

Functional drink containing health extract of citrus peel

The present invention relates to a functional beverage for health improvement manufactured using the extract of citrus fruit, and more particularly, to a functional beverage for health improvement manufactured using bioflavonoids derived from citrus fruit extract or citrus peel.

Citrus peels contain various vitamins, small amounts of limonoids, about 0.01% phenolic and about 1 to 7% bioflavonoids (Ranganna S. et al., Citrus fruits: varieties, chemistry, technology, and quality evaluation.Part II.CRC Crit. Rev. Food Sci.Nutri.Boca Raton , 18 , 313-386, CRC Press (1983)). Among them, bioflavonoids include a wide variety of flavonoids as shown in Table 1 (Horowitz, RM et al., J. org. Chem. , 25 , 2183-2187 (1960)).

As shown in Table 1, hesperidin is found in the extract of orange, lemon, and tangerine, and naringin is present in the extract of grapefruit.

Citrus peels are known to have a variety of pharmacological activities, for example, blood pressure, arteriosclerosis, cardiovascular disease prevention, anti-inflammatory, antioxidant, anticancer, antiviral, capillary potentiation. For example, hesperidin or hesperetine is known to have capillary fortification, reduced permeability, lowered blood pressure, lowered blood cholesterol, antiplatelet aggregation, anti-inflammatory and antiviral effects (Meyer, OC, Angiology). , 45 , 579-584 (1994); Struckmann JR, et al., Angiol., 45 , 419-428 (1994); Matsubara, Y., et al., Japan Organic Synthesis Chem. Association Journal., 52 , 318 -327 (1994. Mar.); Galati EM, et al., Farmaco., 51 (3) , 219-221 (1996, Mar.); Monforte MT., Et al., Farmaco., 50 (9) , 595-599 (1995, Sep.); JP 95-86929; JP 95-86930; Chung, MI, et al., Chin. Pharm. J. (Taipei) ., 46 , 429-437 (1994, Nov.) Galati, EM, et al., Farmaco., 40 (11) , 709-712 (1994, Nov.); and Emim JA, et al., J. Pharm.Pharmacol ., 46 (2), 118-122 (1994)), bioflavonoids are known to have antioxidant, hypotensive, anticancer and antiviral effects (Saija, A., et al., Free Radical Biol. Med., 19 , 481-486 (1995); Mats ubara, Y., et al., supra; Galati EM, et al., supra; Felicia V., et al., Nutr. Cancer, 26 , 167-181 (1996); EP 0352147 A2 (1990.1.24) and Kaul, TN, et al., J. Med. Virol., 15, 71-79 (1985)), naringenin and naringin are known to have cholesterol lowering, anticancer and anti-gastric ulcer (protection from alcohol) actions. (Monforte MT. et al., supra; JP 95-86929; JP 95-86930; Felicia V. et al., Supra; EP 0352147 A2 (1990.1.24); and Martin MJ et al., Pharmacol., 49, 144-150 (1994)).

In addition, citrus rinds and seeds contain limonoids, which have been shown to have anticancer activity (Lam, LKT et al., Inhibition of chemically induced carcinogenesis by citrus limonoids.In Food Phytochemicals for Cancer Prevention, Vol. I, ACS Symposium series No. 546, Huang, MT, O. Osawa, CT Ho, R. Rosen (ed). 1993).

There are many reports that phenolic in citrus fruits has anticancer effect (Stich HF et al., Adv. Exp. Med. , 177 , 1-29 (1984); Stavric B. et al., Effects of flavonoids on mutagenicity and bioavailability of xenobiotics in foods.in Huang MT, CT Ho, CY Lee (eds) (1992): Phenolic compounds in food and their effects on health.II.Antioxidants and cancer prevention.ACS Symposium Series 507, Washington, DC, ACS pp239-249). In addition, phenolic, especially Ella ginsan, toy acid, caffeic acid in the cancer suppressing effect (Castonguay A. Ann. NY Acad. Sci., 686, 177-185 (1993)), and inhibition of cancer generated in the tongue of mice effect (Tanaka T. et al., Carcinogenesis , 14 , 1321-1325 (1993)) has also been reported.

The inventors have already discovered that citrus peel extract, hesperidin or naringin inhibit 3-hydroxy-3-methylglutaryl coeigen (HMG CoA) reductase, an enzyme involved in cholesterol biosynthesis, and acetyl, an enzyme involved in cholesterol absorption. It confirmed that it inhibits the Coyl: cholesterol acyltransferase (ACAT) and filed with Korean Patent Application Nos. 97-55578, 97-55579, 97-55580, respectively. When the citrus peel extract, hesperidin or naringin can be added to various vegetable juices, fruit juices, carbonated drinks, other beverages, the present invention was completed.

It is an object of the present invention to provide a functional beverage for health improvement comprising a citrus peel extract, or a bioflavonoid derived therefrom.

In the present invention, a citrus fruit extract, or a health-improving functional beverage containing hesperidin or naringin, which are separated or commercially available from the same, provides vegetable juice, fruit juice and carbonated beverage.

Citrus fruits in the present invention refers to a mandarin orange, sugar citron, orange, lemon, citrus, grapefruit or citron. In order to obtain citrus peels used in the present invention, citrus fruits must first be produced without pesticide treatment or by using low pollution pesticides or biological control agents that are harmless to humans.

In the present invention, the following methods are used to prepare the citrus peel extract. That is, after drying the citrus peel, 3 to 30 l of 20 to 95% ethanol per kg of dried peel was added, extracted at 25 to 80 ° C. for 1 to 12 hours, and then the extract was filtered and the filtrate was concentrated in vacuo. Extracts can be obtained.

On the other hand, after drying the citrus peel, 0.1 to 2% Ca (OH) ₂ or 5 to 30 L of NaOH per 1 kg of dried peel is added and extracted at 25 to 60 ° C. for 1 to 5 hours, and then the extract is extracted. After filtering, 1N HCl solution was added to the filtrate to adjust the pH of the mixed solution to 4.0 to 7.0, and the resultant precipitate was recovered by drying in a refrigerating chamber for 10 to 48 hours, followed by drying.

The functional foods of vegetable juice, fruit juice and carbonated beverages of the present invention include 200-10000 mg of hesperidin, 200-10000 mg of naringin or 10-100 g of citrus rind extract with respect to 1000 ml of beverage.

The vegetable juice may include carrot juice, and the fruit juice may include grape juice, pineapple juice, apple juice or banana juice, and the carbonated beverage may include coca cola and pepsi.

Ingesting a beverage composition for improving health, comprising the extract of citrus peel, prepared by the method of the present invention, will greatly help our body's defense ability against cardiovascular disease, cancer, virus, inflammation, diabetes, etc. will be.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following Examples are only for illustrating the present invention, and the scope of the present invention is not limited to these.

Example 1 Extraction and Quantitative Methods of Bioflavonoids from Citrus Peel Extracts

The peels of oranges, grapefruits and lemons, which were produced in Jeju, South Korea, were imported from citrus fruits, tangerines, citrons and imported citrus fruits from Jeonnam, and then powdered to a particle size of 100 μm-200 μm. 500 mg of this powder was precisely weighed, and 50 ml of methanol was added thereto, followed by extraction with a water bath at 50 ° C. for 6 hours, followed by cooling, filtration, and addition of methanol to make a 50 ml solution. If necessary, the mixture was diluted to have a concentration within the range used for preparing the calibration curve.

In order to investigate the components of the skin extract obtained as described above, HPLC analysis was performed under the following conditions. 5.0 μl of the sample solution was injected into an HPLC equipped with a Lichrosorb RP-8 column (5 μm, 4 × 250 mm) previously equilibrated with 37% methanol. The temperature of the column was adjusted to 30 ° C. and eluted at a flow rate of 1.0 ml / min using 37% methanol as the mobile phase. Separately, hesperidin and nargine (Sigma Co.) standards were dissolved in methanol and standard solutions were prepared so that the final concentrations were 0.1, 0.2, 0.3, 0.4 and 0.5 mg / ml, respectively, and HPLC was performed under the same conditions. The eluate was detected at 280 nm using a UV-VIS spectrophotometer, and the content was calculated by comparing the area of the HPLC profile of the extract with the calibration curve and the content of hesperidin and naringin in the extract extracted from the extract. The content was as follows.

Content of Hesperidin and Naringin in Various Citrus Peels Citrus varieties Hesperidin Narinjin Orange Tangerine Orange Lemon Lemon Tangerine Grapefruit -0.402.101.402.10-0.80 3.403.90 Micro Trace Micro Trace 4.700.80

Example 2: Preparation of Citrus Peel Extract

(1) Method using ethanol

After drying the citrus peel from Jeju Island, 5 l of 30% ethanol solution was added to 500 g of dried citrus peel and extracted at 60 ° C. for 5 hours. The extract was filtered using cotton cloth, and the filtrate was concentrated in vacuo to obtain 190 g of syrupy extract. As a result of measuring the content of hesperidin in the extract in the same manner as in Example 1, 190 g of hesperidin was contained in the extract. As a result of analyzing the components of the extract using HPLC, it consisted of trace components such as water 65%, sugar 28% (11% fructose, glucose 11%, sucrose 6%), hesperidin 2.7% and other ash.

(2) Method using Ca (OH) ₂

After drying the citrus peel from Jeju Island, 5 liters of 0.5% Ca (OH) ₂ solution was added to 500 g of the dried citrus peel and stirred for 1 hour while stirring at room temperature. The extract was filtered with cotton fabric and 1N HCl solution was added to the filtrate to make the pH of the mixed solution 4.0 to 4.5, or 6.80 to 7.0, and then left in a 5 ° C. cold room for 24 hours. At this time, the resulting precipitate was recovered and dried to obtain 5 g and 10 g of dried powder, respectively. The precipitated powder was quantitatively analyzed by HPLC as in Example 1, containing 3.2 g and 6.55 g of hesperidin, respectively, and the purity of hesperidin was 64% and 65%, respectively.

(3) Method using NaOH

After drying the citrus peel from Jeju Island, 5 l of 0.5% NaOH was added to 500 g of the dried citrus peel and stirred at room temperature for 1 hour, followed by filtration with cotton fabric. 1N HCl solution was added to the filtrate to bring the pH of the mixed solution to 4.0-4.5 or 6.8-7.0, and then left in a 5 ° C. refrigerator for 24 hours. At this time, the resulting precipitate was recovered and dried to obtain 44 g and 49 g of dry powder, respectively. 13.6 g and 9.8 g of hesperidin, respectively, were recovered from the dry powder by HPLC (purity: 31%, 20%, respectively).

Example 3: Oral Toxicity Test of Citrus Peel Extract

Specific pathogens free of 7-8 weeks of age, 6 females weighing 25-29 g and 6 males weighing 34-38 g, temperature 22 ± 1 ° C, humidity 55 ± 5%, illumination 12L It was bred in the animal room of / 12D. Mice were allowed to acclimate for about a week before being used in the experiment. Feed for experimental animals (Jeil Jedang Co., Ltd., mice and rats) and drinking water were supplied after sterilization and free ingestion.

Citrus peel extract obtained in (1) of Example 2 was prepared at a concentration of 100 mg / ml using 0.5% Tween 80 as a solvent, followed by oral administration of 0.2 ml per 20 g of mouse body weight. Samples were administered orally once and observed for side effects or lethality for 10 days after administration. That is, on the day of dosing, changes in general symptoms and the presence or absence of dead animals were observed at least once in the morning, at least once every afternoon from 1 hour, 4 hours, 8 hours, 12 hours after the administration and from the next day until the 10th. In addition, the animals were killed and dissected at 10 days of administration, and the internal organs were visually examined. Changes in body weight were measured at daily intervals from the day of administration, and the weight loss of the animals by the citrus peel extract was observed.

The purpose of this experiment was to provide information on available doses in general pharmacology and drug efficacy tests and to derive basic data on toxicity by identifying the degree of acute toxicity in the oral route of citrus peel extracts. The result was obtained.

In the case of acute oral toxicity, no lethal animals were observed for 10 days at the dose of 1000 mg / kg citrus peel extract. An autopsy of the surviving animals was performed 10 days later, and there were no gross lesions, and normal body weight gain was observed without any weight loss for 10 days from the day after the administration.

In conclusion, the citrus peel extract was not toxic upon oral administration.

Example 4 Dietary Composition of Experimental Animals and Animal Administration of Citrus Peel Extracts

20 Sprague Dawley rats, 3 weeks old, were purchased from the Korean Experimental Animal Center in Eumseong-gun, Chungbuk, Korea, and were bred for one week with experimental animal feed and fertilized at 4 weeks of age (weight: 90-110 g). It was divided into two groups by randomized block design. Two groups of rats each had two different high-cholesterol diets, one containing 1% cholesterol (control) in the normal diet (AIN-76 experimental animal diet) and one percent cholesterol and 5% citrus peel The diet containing each obtained in Example 1 (1) was consumed. The composition of the diets ingested in both groups of rats is shown in Table 3.

Dietary group Control Citrus peel extract group Casein 20 20 D, L-methionine 0.3 0.3 Corn starch 15 15 Sucrose 49 49 Cellulose powder 5 0 Mineral mixtures 3.5 3.5 Vitamin mixtures One One Choline Citrate 0.2 0.2 Corn oil 5 5 cholesterol One One Citrus Peel Extract - 5 gun 100 100

All diets correspond to the high cholesterol diet (1%, wt / wt), and the cellulose, mineral and vitamin mixtures in the AIN-76 diet, the laboratory rat's laboratory diet, were obtained from the US-based TEKLAD premier, Madison. , Wisconsin).

Rats of both diets were allowed to eat the diet freely with water for 6 weeks, and their dietary intake was recorded daily and weighed every 7 days. After the animal breeding was completed, the data on the breeding journals were analyzed, and there was no significant difference between the two groups in terms of dietary intake and weight gain.

Example 5 Determination of Total Cholesterol, HDL-Cholesterol and Neutral Lipid Contents in Experimental Animals

The effects of experimental diet on blood cholesterol and triglyceride content in rats were investigated as follows.

Blood samples were taken from the rats of both diets, from which plasma H6DL fractions were separated using HDL-cholesterol reagent (Sigma Chemical Co. Cat. No. 352-3) including dextran-sulfate. Total cholesterol and HDL-cholesterol concentrations were measured by enzymatic coloration using Sigma's Total Cholesterol Kit (Cat. No. 352-100) (Allain et al., Clin. Chem., 20 , 470-475 (1974)). It was. Neutral lipid concentrations were measured using Sigma Diagnostic Kit (Cat. No. 339-50) (Bucolo, G. and David, H., Clin. Chem., 19 , 476-482 (1973)). As a result, the plasma total cholesterol concentration was 20% decreased in the citrus peel extract group compared with the control group, but there was no significant change in the HDL or triglyceride concentration.

Example 6 Preparation of Functional Drink for Health Improvement Using Citrus Peel Extract

Functional beverages for improving health comprising the tangerine peel extract prepared in Example (1) of Example 2 were prepared as follows.

(1) vegetable juice production

Hesperidin or naringin 200 mg to 10000 mg, or 10-100 g of citrus peel extract were added to 1000 ml of various vegetable juices to prepare functional vegetable juices.

(2) fruit juice production

Hesperidin or naringin 200 mg to 10000 mg, citrus peel extract 10 to 100 g was added to 1000 ml of grape, pineapple, apple or banana fruit juice to prepare a functional fruit juice.

(3) carbonated beverage manufacturing

200 mg to 10000 mg of hesperidin or naringin and 1 to 100 g of citrus peel extract were added to 1000 ml of Coca Cola or Pepsi Cola to prepare a functional carbonated beverage.

The functional beverage for health improvement prepared by adding the extract of citrus peel containing hesperidin or naringin to various beverages may exhibit effects such as antioxidant activity, lowering blood pressure, lowering blood cholesterol, and anticancer action.

Claims (3)

A functional beverage for health improvement prepared by adding 200-10000 mg of hesperidin, 200-10000 mg of naringin, or 10-100 g of citrus rind extract with respect to 1000 ml of beverage. The method of claim 1, The citrus fruit is a functional drink of mandarin orange, sugar, orange, lemon, citrus, grapefruit or citron. The method of claim 1, Functional drink for health improvement, characterized in that the beverage is vegetable juice, fruit juice or carbonated beverage.