KR100857003B1 - A method for preparing extract of small molecular from citrus peel and the extract of small molecular - Google Patents

Abstract

A method of manufacturing citrus peel extract having a low molecular weight is provided to obtain citrus peel extract containing a physiologically active substance such as hesperetin which is easily digested and absorbed into the body in large quantities by decomposing the cell wall structure of the citrus peel. Citrus peel extract having a low molecular weight is prepared by the steps of: washing citrus peel with water and then ozone water, drying at room temperature and coarsely crushing; adding 5 to 10 times the weight of water to coarsely crushed citrus peel and heating at 90 to 120deg.C for 10 to 14hr; adding probiotics containing 1.0x10^6 to 1.0x10^9/g Streptococcus faecalis, Bacillus mesentericus and Clostridium butyricum on a dry basis to the citrus peel extract and fermenting at 30 to 38deg.C for 4 to 6hr; adding water to the fermented citrus peel liquid and extracting.

Description

A method for preparing extract of small molecular from citrus peel and the extract of small molecular}

1 is a chromatogram obtained by measuring the molecular weight distribution by a size exclusion column high performance liquid chromatography.

2 is a graph showing the relationship between the molecular weight and the elution time based on the chromatogram of the molecular weight standard.

The present invention relates to a method for low molecular weight extraction of constituents of dermis so as to effectively utilize effective ingredients such as hesperidin contained in citrus dermis (dermis) and dermis extract thus obtained.

Citrus fruits, such as Wenzhou citrus, mandarin orange, sugar citron, citron, blueberry, and tangerine, are mainly used only for the flesh, such as steaming to eliminate juice and to eliminate sour taste, so that a large amount of skin (dermis) is generated. Most of these citrus dermis are disposed of, and research on how to use them is required. In Korean Patent Publication No. 1992-0004229, a method of manufacturing feed using citrus peels is disclosed. In Korean Patent Publication No. 10-0381726, a citrus fermentation solution as a feed additive for aquaculture, and in Korean Patent Publication No. 10-0469831, citrus charcoal and its It describes a manufacturing method. These methods focused on the use of the dermis for non-edible uses such as animal feed.

Citrus dermis, however, contains pectin-containing dietary fiber, carotenoids, vitamins and flavonoids (Horowitz et al., Flavonoids of Citrus. IV.Isolation of some aglycones from the Lemon ( Citrus) limon ). J. Org. Chem. 25: 2183-8187, 1960) and many other bioactive substances useful for the human body. Dietary fiber in the dermis is known to help prevent atherosclerosis, hyperlipidemia and diabetes by lowering the absorption of fat, cholesterol and sugar in the intestine when a person consumes a sufficient amount. In addition, several physiological activities of dermal-derived flavonoids have recently been reported. In particular, hesperidin, also called vitamin P, has been reported to be effective in protecting capillaries and preventing atherosclerosis and hypertension. In addition, nobiletin of flavonoids, beta-cryptoxanthin and coumarin of carotenoids have been reported to have anticancer activity, and high content of vitamin C in the dermis has anti-aging effect. Pectin amino acids, limonene (limonene) is known to have a hair growth effect. As the contents of the active ingredient having various pharmacologically active activities were found to be higher in the dermis than the citrus pulp, more studies were conducted on the use of the dermis for food and the efficacy of the active ingredient derived from the citrus dermis. It is done.

In Korean Patent Publication No. 10-0213895, registered in 1999, hesperidin or naringin, which is known to be effective in preventing and treating cardiovascular diseases, was isolated from citrus dermis, and Murakami et al. (Suppressive effects of citrus fruits on free radical generation and nobiletin and anti-inflammatory polymethoxyflavonoid.BioFactors, in press.2000) Evaluation of the anti-allergic activity of Citrus unshiu using rat basophilic leukemia RBL-2H3 cells as well as basophils of patients with seasonal allergic rhinitis to pollen.International Journal of Molecular Medicine 17: 511-515, 2006). In addition, Korean Patent No. 0508897 discloses a citrus fruit composition in which the content of the active flavonoid compound is increased by converting a glycoform-inactive intact full-lavonoid compound into an aglycone-type active flavonoid compound by adding glycolytic enzymes such as cellulase to citrus fruits. It describes the preparation method of. As such, the methods of using the dermis for food have been mostly extracted or used only with specific substances or supplemented with the flesh.

Korean Patent No. 0665016, recently registered, uses a method of manufacturing dermis tea using organic citrus fruits, which are semi-dried citrus dermis, crushed to a certain size or less, and manufactured for multi-tube and tea bags so that they can be consumed in hot water and consumed. Suggesting. However, the use of tea as a tea soaked in hot water for such a short time has a fundamental limitation in terms of extracting the active ingredient. In particular, the outer wall of plants such as the dermis is composed of components such as pectin, cellulose, lignin, beta-1,4-glucan, which are difficult to degrade. However, since bioactive substances exist inside these outer walls, it is difficult to extract them by simple hydrothermal extraction. In addition, some of the extracted bioactive substances are also in the form of polymer polysaccharides or glycosides, so if there are no enzymes or microorganisms capable of degrading them in the body, they will not be decomposed, and thus the absorption rate will be degraded.

An object of the present invention is to provide a method for low molecular weight extraction of constituents of dermis so that effective ingredients such as hesperidin contained in citrus dermis (dermis) can be effectively used. To this end, in the present invention, Streptococcus paecalis ( Streptococcus faecalis ); Bacillus mesentericus ; Probiotics consisting of Clostridium butyricum and their sugar transfer enzymes are used to decompose the dermal wall structure to make it easier to extract various bioactive substances present in the wall. Effectively extract the active ingredient from the dermis by the method of hot water extraction over. In addition, the bioactive substances extracted through such a process, in particular, glycosides such as hesperidin are decomposed into sugar and aglycone form, which is changed to a state in which digestion and absorption are easy in the human body.

In another aspect, the present invention provides a dermis extract containing a large amount of bioactive substances such as hesperidin or hesperetin (hesperetin), which is extracted in this manner, can be used in various applications such as health food, beverages, cosmetics, etc. The purpose is to make it.

Other objects and advantages of the present invention will be described below and will be better understood by practice of the present invention.

In the present invention,
(a) adding water and ripening the coarsely ground dermis of citrus fruits;
(b) Probiotics containing 1.0 × 10 ^{6 to} 1.0 × 10 ⁹ / g of Streptococcus faecalis , Bacillus mesentericus and Clostridium butyricum on a dry basis, respectively Adding to the cooked dermis obtained in the step (a) and fermented to obtain a dermal fermentation broth;
(c) adding water to the dermal fermentation broth obtained in step (b) and heating the extract to take the solution. Provided is a method for producing a low molecular weight dermis extract comprising the.

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In the present invention,

There is provided a health food containing the low molecular weight dermis extract prepared by the method as an active ingredient.

In the present invention, "citrus fruits" is meant to include all the citrus, mandarin orange, sugar citron, citron, blueberry, tangerine, products and the like.

"Dermis" in the present invention means mature peel, the fruit of the citrus is removed.

1. The steps to ripen citrus dermis

Water is added to the citrus dermis, heated to cook, and cooled to room temperature. Preferably, water is added to the coarsely pulverized dermis, heated to 90 to 110 ° C. for 1 to 2 hours, cooked sufficiently, and then slowly cooled to 20 to 40 ° C. At this time, More preferably, about 2 to 5 times of water is added and heated. When the dermis is cooked and fermented, the solid wall structure of the dermis is decomposed, so that it is easy to extract various bioactive substances present in the wall.

Prior to performing this step, a pretreatment is preferably carried out to wash and dry the surface of the dermis. Preferably, the dermis of the citrus or pulp removed state is first immersed in fresh water for about 1 hour and washed with water to remove foreign substances on the surface. More preferably, citrus fruits or dermis from which foreign substances have been removed are washed again with ozone water and dried at room temperature. The ozone water generates active oxygen with strong sterilization, deodorization and high reactivity with metals to sterilize microorganisms on the dermal surface and remove residual pesticide components. After washing with water and drying, citrus fruits are separated into pulp and dermis and then coarsely pulverized.

2. Fermentation stage

While maintaining the cooked dermal solution obtained in the above step at 20 ~ 40 ℃, Streptococcus faecalis , Bacillus mesentericus , Clostridium butyricum on the basis of the dry amount Probiotics containing 1.0 × 10 ^{6 to} 1.0 × 10 ⁹ / g, respectively, are added to the cooked dermis obtained in the above step and fermented. Preferably, 0.1-0.2 g of fermented 1 liter of cooked dermis is added and fermented for 2 to 8 hours, more preferably at 4 to 6 hours at 30 to 38 ° C. In one embodiment of the present invention, as a probiotics, "Probiotics Powder Complex" of TOA PHARMACEUTICAL, Japan was used, but the available probiotics are not limited thereto, and the probiotic surface containing at least 1.0 × 10 ⁶ / g of the three strains, respectively. The probiotics preferably contain 1.0 × 10 ^{6 to} 1.0 × 10 ⁹ / g, and more preferably 1.0 × 10 ^{7 to} 1.0 × 10 ⁸ / g, respectively, of the microorganisms. On the basis of quantity, 0.1 to 0.2 g of probiotics is added to the dermis obtained in the above step, preferably 0.12 to 0.18 g, and the probiotic preparation contains enzymes, especially sugar transfer enzymes, of the microorganisms contained therein. Since the enzyme is also produced in the process, sugar transfer with the cells of the microorganisms contained, even if the sugar transfer enzyme is not used separately There is an effect that cows put together. Fermentation time is typically 2-8 hours, preferably 4~6 hours suitable one, it is possible to further extend the time or speed, as needed.

 Through this fermentation process, the wall structure of the dermis, which is composed of components such as pectin, cellulose, lignin, beta-1,4-glucan, is decomposed or Changed to a looser structure, it is easier to extract various bioactive substances such as hesperidin inside the wall, and through this fermentation process, biomolecules that existed in the dermis in the form of glycosides such as polymer polysaccharide and hesperidin have low molecular weight. It can be broken down into sugar and aglycone, which can be easily digested and absorbed by the human body. After this process and undergoing the hot water extraction process of the next step, preferably 70% or more, more preferably 80% or more of the components contained in the whole dermis extract is a low molecular state of less than 1000 molecular weight.

3. Extraction step

After the fermentation step is completed, water is added and extracted by heating for 5 to 20 hours. Preferably, 5 to 10 times of water is added and heated at 90 to 120 ° C. for 10 to 14 hours to extract effective bioactive substances. After extraction, cool to room temperature and take only the supernatant or filter the solution. Through this extraction step, the sterilization of the fermentation microorganism and the inactivation of the enzyme are also made at the same time. The extract obtained in this step can be used as it is, or concentrated or dry powdered. Concentration or dry powdering may be applied with known conventional methods used in the food processing field. The dermis extract obtained according to the present invention can be used in various forms in various fields such as beverages, health foods, cosmetics, etc. In particular, it can be used as a raw material for health functional foods and cosmetics of capsule or tablet type in a powdered state.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, the scope of the present invention is not limited by the following examples, and those skilled in the art to which the present invention pertains should be within the equivalent scope of the technical concept of the present invention and the claims to be described below. Of course, various modifications and variations are possible.

Example  One

Pretreatment and dermal ripening steps

3 kg of Wenju mandarin persimmon, 1 kg of sugar citron, and 11 kg of product, which are sold in Jeju Island, were immersed in fresh water for 1 hour to remove foreign substances, and the freshly removed citrus fruits were first immersed in ozone water for 2 hours. , Fertilizer components were removed, and immersed in clean ozone water for 3 hours to remove foreign substances, residual pesticides and fertilizer components as much as possible. Washed and sterilized citrus fruits were air-dried at room temperature until there was no moisture on the fruit surface. The dried citrus fruits were separated into pulp and dermis, and the separated dermis was coarsely ground using a grinder. Four times of water was added to the pulverized dermis thus heated to 105 ° C. for 90 minutes, and then slowly cooled to room temperature to maintain about 30 ° C.

Example  2

Fermentation stage

60L of cooked dermis obtained in the above step was prepared by Toa Pharmaceutical Co., Ltd. (TOBI PHARMACEUTICAL Co., Ltd.) Probiotics Powder Complex: Streptococcus faecalis T-110; Bacillus mesentericus TO-A; Clostridium butyricum TO-A, the number of cells per g of 2.4 × 10 ⁸ CFU (colony forming unit), 6.7 × 10 ⁸ CFU, 1.1 × 10 ⁷ CFU) was added 8g and fermented at 35 ℃ for 5 hours.

Example  3

Extraction step

10 times of water was added to the dermal fermentation broth obtained in Example 2 and extracted by heating at 100 ° C for 30 hours. After extraction, the mixture was cooled to room temperature and the supernatant was taken as the dermis extract of the present invention.

Experimental Example

The molecular weight distribution of the components contained in the dermis extract obtained in Example 3 was measured using a size exclusion chromatography using TSKgel GMPWXL coulumn. The experiment was commissioned by the Japan Food Analysis Center.

1. Test Method

1) Preparation of Standard Solution

A molecular weight standard (such as pullulan) as shown in Table 1 was dissolved in 0.1 mol / L sodium acetate solution to obtain a 0.05W / V% solution.

*: Shodex standard P-82 [Showa Major Co., Ltd.]

2) Preparation of test solution

0.2 g of the dermis extract obtained in Example 3 was taken and placed in 10 ml of 0.1 mol / L sodium acetate solution. After standing at room temperature overnight, it was bathed in boiling water for 10 minutes. After cooling at room temperature, the filtrate obtained by filtration with a membrane filter of pore-size 0.45 탆 was used as a test solution.

3) Molecular weight distribution measurement

Standard and test solutions were injected into a high performance liquid chromatography (HPLC) device using a size exclusion column. The results obtained were analyzed using a 480II data station GPC (gel permeation chromatography) program (SYSTEM INSTRUMENTS Co. ltd.).

2. Results

The result of measuring the molecular weight distribution is shown in Table 2 below. The obtained chromatogram is shown in FIG. 2 shows the graph of the relationship between the molecular weight and the elution time based on the chromatogram of the molecular weight standard. As a result of the experiment, most of the components up to 84% were found to have a molecular weight of 1,000 or less.

According to the present invention, through the microbial fermentation and hot water extraction process, the wall structure of the dermis can be decomposed to easily extract various bioactive substances present in the wall. The dermis extract prepared according to the present invention contains various bioactive substances derived from dermis such as pectin, carotenoids, flavonoids, vitamins, etc., and thus can be utilized in various fields such as health food, medicine, cosmetics, agriculture and livestock, etc. It can be effectively used in beverages, health foods and cosmetics.

Claims (6)

(a) adding water and ripening the coarsely ground dermis of citrus fruits; (b) Probiotics containing 1.0 × 10 ^{6 to} 1.0 × 10 ⁹ / g of Streptococcus faecalis , Bacillus mesentericus and Clostridium butyricum on a dry basis, respectively Adding to the cooked dermis obtained in the step (a) and fermented to obtain a dermal fermentation broth; (c) adding water to the dermal fermentation broth obtained in step (b) and heating the extract to take the solution. The method for preparing a low molecular weight dermis extract of claim 1, further comprising a pretreatment step of washing the dermis with water, washing with ozone again, drying at room temperature, and then coarsely pulverizing. The method of claim 1, wherein the step of extracting heat by adding 5 to 10 times of water to the dermal fermentation broth and extracting by heating for 10 to 14 hours at 90 to 120 ℃. According to claim 1, wherein step (b) is Streptococcus faecalis ( Streptococcus faecalis ); Bacillus mesentericus ; Probiotics containing 1.0 × 10 ^{7 to} 1.0 × 10 ⁹ / g of Clostridium butyricum , respectively, based on the dry weight of 0.1-0.2 g was added to 1 L of the cooked dermis obtained in step (a). Method for producing a low molecular weight dermis extract, characterized in that the fermentation for 2 to 8 hours. The method of claim 4, wherein the fermentation is a method for producing a low molecular weight dermis extract, characterized in that the fermentation for 4 to 6 hours at 30 to 38 ℃. Health food containing the low molecular weight dermis extract prepared by the method of claim 1 as an active ingredient.

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