WO2008127063A1

WO2008127063A1 - A method for preparing the inclusion complex of ginseng extract with gamma-cyclodextrin, and the composition comprising the same

Info

Publication number: WO2008127063A1
Application number: PCT/KR2008/002112
Authority: WO
Inventors: Seung Hyun Lee; Heui Jong Yu; Nam Suk Cho; Ji Ho Park; Tae Hyun Kim; Ki Ho Kim; Sung Ku Lee
Original assignee: Bioland Ltd.
Priority date: 2007-04-16
Filing date: 2008-04-15
Publication date: 2008-10-23
Also published as: KR100865047B1; KR20080093186A

Abstract

The present invention provides a method for preparing the inclusion complex of ginseng extract with gamma-cyclodextrin and, the composition comprising the same, specifically, a method for preparing the inclusion complex of ginseng extract with gamma-cyclodextrin providing favorable advantages such as the removal of bitter component of ginseng extract with holding the innate flavor thereof, the increase of the solubility and the stability of ginseng extract, which results in transparent liquid type product with high quality.

Description

A METHOD FOR PREPARING THE INCLUSION COMPLEX

OF GINSENG EXTRACT WITH GAMMA-CYCLODEXTRIN,

AND THE COMPOSITION COMPRISING THE SAME

Technical Field

[1] The present invention relates to a method for preparing the inclusion complex of ginseng extract with gamma-cyclodextrin, and the composition comprising the same, specifically, a method for preparing the inclusion complex of ginseng extract with gamma-cyclodextrin providing favorable advantages such as the removal of bitter component of ginseng extract with holding the innate flavor thereof, the increase of the solubility and the stability of ginseng extract, which results in transparent liquid type product with high quality.

[2]

Background Art

[3] Ginseng products have been widely used for improving human health for long years ago in Asian countries and the ginseng extract contains various potent and pharmacologically active substances, such as high amount of various ginsenosides, such as gin- senoside RbI, Rb2, Rc, Rd, RgI, Re etc, showing various pharmacological activities such as immuno-potentiating activity, ant-inflammatory activity, anti-allergy activity, anti-cancer activity, blood pressure lowering activity, cholesterol lowering activity, anti-thrombic activity, etc.

[4] However, the innate bitter tastes of ginseng product have been reported to give rise to several problems, for example, a limited use in the amount of dosage, difficulty in applying to various formulations such as beverage, extract etc., particularly, the formulation for Md, in spite of its good functionality.

[5] There have been tried to solve the problems due to bitter tastes of ginseng products till now. Among the method to solve the problems, the method for scavenging bitter taste in oral formulation and the method for reducing the bitter taste using other components, for example, the reducing method using aminoalkylmethacrylate copolymer and/or glutamates disclosed in Korea Patent Publication No. 10-2001-77882; the reducing method using flavoring agent such as aspartam disclosed in Japanese Patent Publication No. 1990-56416 etc., have been tried till now. However, the methods could not afford satisfactory effect on masking bitter taste of ginseng extract and the more efficient methods have been still needed till now.

[6]

[7] Ganma-cyclodextrin, a malto-oligosaccharide, consists of eight glucoses forming cyclic donut-like ring linked with alpha- 1, 4-linkage molecules and shows ambivalent property, i.e., hydrophobic interior area due to hydrogen bonding between the hydroxyl groups in C-2 and C- 3 and hydrophilic exterior area due to polar hydroxyl groups oriented to outer direction, of which property enable to enhance the solubility of hydrophobic substance or unstable substances by forming an inclusive substance. Moreover, the bitter tastes or unpleasant scent can be removed through the inclusive formation with a part or all of the organic substance showing pleasant taste and favor.

[8] In particular, the solubility at 25⁰C of gamma-cyclodextrin (30.0g/100ml) is higher than those of alpha-cyclodextrin (16.0g/100ml) and beta-cyclodextrin (1.85g/100ml). Furthermore, the internal width surrounded with glucoses of gamma-cyclodextrin (8.3A) is greater than those of alpha-cyclodextrin (5.3A) and beta-cyclodextrin (6.5A). Therefore, gamma-cyclodextrin is the most efficient for forming the inclusive useful in removing unspecific bitter ingredients in plant extract (Pro. Biochem., 39, ppl033-1046, 2003; Chem. Rev., 98, ppl743-1753, 1988).

[9]

[10] However, there has been not reported or disclosed on the use of gamma-cyclodextrin in the formation with ginseng extract to remove the bitter taste thereof in any of the above cited literatures, the disclosures of which are incorporated herein by reference.

[11] The inventors of the present invention have investigated an removing effect of the inclusion complex of ginseng extract with gamma-cyclodextrin comparing with those of alpha-cyclodextrin and beta-cyclodetrin through various experiments, i.e., a sensory test, transparency determination test, stability determination test, NMR determination etc. and finally completed present invention by confirming that a gamma-cyclodextrin effectively scavenged the bitter taste by forming the inclusion complex with bitter components of ginseng extract with holding innate flavor of itself as well as enhanced the solubility and the stability of ginseng extract, and the transparency of liquid formulation containing ginseng extract.

[12]

[13] These and other objects of the present invention will become apparent from the detailed disclosure of the present invention provided hereinafter.

[14] Disclosure of Invention Technical Problem

[15] The present invention provides a method for preparing the inclusion complex of ginseng extract with gamma-cyclodextrin.

[16] Also, the present invention provides a food or food additive composition comprising the inventive inclusion complex.

[17] The present invention provides a health food or food additives comprising the above- described inventive inclusion complex, together with a sitologically acceptable additive for reinforcing immune resisting power.

[18]

Technical Solution

[19] Accordingly, it is an object of the present invention to provide a method for preparing the inclusion complex of ginseng extract with gamma-cyclodextrin, and the composition comprising the same.

[20]

[21] Hereinafter, the present invention is described in detail.

[22]

[23] Specifically, the present invention provides a method for preparing the inclusion complex of ginseng extract with garrma-cyclodextrin comprising the steps consisting of: extracting a ginseng material with 1 to 20-fold volume of water, ethanol or the mixture thereof and concentrating the filtrate with filtering the residue to obtain ginseng extract at the 1^st step; adding distilled water to the extract to obtain diluted solution at the 2^nd step; and adding gamma-cyclodextrin thereto with stirring to perform inclusive reaction at the 3^rd step.

[24]

[25] IVbre specifically, The present invention provides a method for preparing the inclusion complex of ginseng extract with garrma-cyclodextrin comprising the steps consisting of: extracting a ginseng material with 3 to 10-fold volume, preferably, 5 to 7-fold volume of water, ethanol or the mixture thereof, preferably, the mixture solution of water and ethanol, more preferably, 40-80 v/v%, most preferably, 60-70 v/v% mixture solution of water and ethanol for the period ranging from 1 hour to 10 hours, preferably, 2 to 5 hours, once to ten times, preferably, 4 to 6 times at the temperature ranging from 70 to 100Q preferably, from 90 to 95⁰C with a hot- water extraction, ul- trasonication extraction or reflux extraction, preferably, reflux extraction, filtering the solution to obtain filtrate and concentrating the filtrate with vacuum at the temperature ranging from 40 to 8(JC preferably, from 60 to 70C to obtain ginseng extract at the 1 ^st step; adding distilled water to the extract to the extent that the amount of solid phase reaches to the range from 10 to 80 w/v%, preferably, 20 to 40 w/v% to obtain diluted solution at the 2^nd step; and adding 0.1 to 99.9 w/w%, preferably, 1 to 40 w/w%, more preferably, 3 to 30 w/w% gamma-cyclodextrin based on the amount of solid phase thereto with stirring at the temperature ranging from 40 to 120C preferably, from 60 to 9OC more preferably, from 80 to 9OC for the period ranging from 0.5 hour to 6 hours, preferably, 1 to 3 hours to perform inclusive reaction at the 3^rd step.

[26] The term "ginseng material" disclosed herein comprises Panax ginseng, Panax quinquefolia, Panax notoginseng, Panax japonica, Panax trifolia, Panax pseudoginseng, Panax vietnamensis, Panax elegatior, Panax wangianus or Panax bipin- ratifidus in the form of a red ginseng, white ginseng, black ginseng, unhusked ginseng, husked ginseng, wild ginseng or cultivated ginseng, etc., preferably, red ginseng.

[27] The term "the inclusion complex of ginseng extract with gamma-cyclodextrin" disclosed herein may be provided with the form of concentrated solution or dried powder.

[28]

[29] To investigate the removing effect of the inclusion complex of ginseng extract with gamma-cyclodextrin prepared by the above-described method comparing with those of alpha-cyclodextrin and beta-cyclodextrin, present inventors performed various experiments, i.e., a sensory test, transparency determination test, stability determination test, NMR determination etc. and confirmed that a garrma-cyclodextrin effectively scavenged the bitter taste by forming the inclusion complex with bitter components of ginseng extract with holding innate flavor of itself as well as enhanced the solubility of ginseng extract, the stability of ginseng extract and the transmittance of liquid formulation containing ginseng extract.

[30]

[31] The present invention provides a food or food additive composition comprising the inventive inclusion complex prepared by the above-described method.

[32]

[33] The inventive composition may additionally comprise one or more than one of organic acid, such as citric acid, funaric acid, adipic acid, lactic acid, malic acid; phosphate, such as phosphate, sodiun phosphate, potassiun phosphate, acid pyrophosphate, polyphosphate; natural anti-oxidants, such as polyphenol, catechin, α- tocopherol, rosemary extract, vitamin C, green tea extract, licorice root extract, chitosan, tannic acid, phytic acid etc.

[34]

[35] The above-described inclusion complex of the present composition may be 20 to 90

% high concentrated liquid, power, or granule.

[36] Similarly, the above-described inclusion complex of the present composition can comprise additionally one or more than one of lactose, casein, dextrose, glucose, sucrose and sorbitol.

[37]

[38] Also, in the present invention, there is also provided a using method of the food additives such as sterilizer, spice, seasoning, various nutrients, vitamin, a mineral or an electrolyte, synthetic flavoring agent, a coloring agent and improving agent in case of cheese chocolate et al., pectic acid and the salt thereof, alginic acid and the salt thereof, organic acid, protective colloidal adhesive, pH controlling agent, stabilizer, a preservative, glycerin, alcohol, carbonizing agent used in carbonate beverage et al., or as essential component of food materials.

[39]

[40] Wherein the food additives can be added to food by deposition, spray, or mixing the ratio of the additives is not so important but is generally range from about 0.01 to 20 w/w % per 100 w/w % present composition. Examples of addable food comprising aforementioned extract therein are.

[41] It is another object of the present invention to provide a health food or food additives comprising the above-described inventive inclusion complex, together with a sito- logically acceptable additive for reinforcing immune resisting power.

[42]

[43] The term "sitologically acceptable additive" described herein may be any substance the intended use which results or may reasonably be expected to result-directly or in- directly-in its becoming a component or otherwise affecting the characteristics of any food for example, thickening agent, maturing agent, bleaching agent, sequesterants, humectant, anticaking agent, clarifying agents, curing agent, emulsifier, stabilizer, thickner, bases and acid, foaming agents, nutrients, coloring agent, flavoring agent, sweetner, preservative agent, antioxidant, etc., which shall be explained in detail as follows.

[44] If a substance is added to a food for a specific purpose in that food, it is referred to as a direct additive and indirect food additives are those that become part of the food in trace amounts due to its packaging, storage or other handling.

[45]

[46] The above-described health food may comprises at least one combination selected from the group consisting of the aloe vera gel extract, green tea leave extract, mugwort extract, pine needles extract, asparagus extract and dropwort extract as a main component; at least one combination selected from the group consisting of Acanthopanacis cortex extract, Chinese quince extract, ginger extract, onion extract; and vitamin mixture comprising vitamin C, B _l5 B₂, B₃ and B₁₂ and folic acid for reinforcing immune resisting power.

[47]

[48] The above described health food comprises confectionary such as chewing gun, candy, biscuits, chocolate, ice cream, etc., meat processed foodstuff, fish processed foodstuff, bean-curd product, jelly product, edible oil, noodle product, beverages, tea product, heath care food, nutrient food, seasoning foodstuff, ginseng product, kimchi or salted food, dried meat or fish food and the like.

[49] The health food of the present invention comprises the above inclusion complex as

0.01 to 95 %, preferably 1 to 80 % by weight based on the total weight of the composition.

[50]

[51] The above-described health food comprise health food, health beverage etc., and may be used as a form of powder, granule, tablet, liquid, chewing tablet, capsule, beverage etc.

[52] To develop for health food, examples of addable food comprising the above described inventive inclusion complex of the present invention are e.g., various food, beverage, gum, vitamin complex, health improving food and the like, and can be used as powder, granule, tablet, chewing tablet, capsule or beverage etc.

[53]

[54] The above-described composition therein can be added to food, additive or beverage, wherein the amount of above described inclusion complex in food or beverage may generally range from about 0.1 to 95 w/w %, preferably 1 to 80 w/w % of total weight of food for the health food composition and 1 to 30 g, preferably 3 to 10 g on the ratio of 10Om^ of the health beverage composition.

[55]

[56] Providing that the health beverage composition of present invention contains above described inclusion complex as an essential component in the indicated ratio, there is no particular limitation on the other liquid component, wherein the other component can be various deodorant or natural carbohydrate etc. such as conventional beverage. Examples of aforementioned natural carbohydrate are monosaccharide such as glucose, fructose etc.; disaccharide such as maltose, sucrose etc.; conventional sugar such as dextrin, cyclodextrin; and sugar alcohol such as xylitol, and erythritol etc. As the other deodorant than aforementioned ones, natural deodorant such as taumatin, stevia extract such as levaudioside A, glycyrrhizin et al., and synthetic deodorant such as saccharin, aspartam et al., may be useful favorably. The amount of above described natural carbohydrate is generally ranges from about 1 to 20 g, preferably 5 to 12 g in the ratio of 100 ml of present beverage composition.

[57]

[58] The other components than aforementioned composition are various nutrients, a vitamin, a mineral or an electrolyte, synthetic flavoring agent, a coloring agent and improving agent in case of cheese, chocolate et al., pectic acid and the salt thereof, alginic acid and the salt thereof, organic acid, protective colloidal adhesive, pH controlling agent, stabilizer, a preservative, glycerin, alcohol, carbonizing agent used in carbonate beverage et al. The other component than aforementioned ones may be fruit juice for preparing natural fruit juice, fruit juice beverage and vegetable beverage, wherein the component can be used independently or in combination. The ratio of the components is not so important but is generally range from about 0 to 20 w/w % per 100 w/w % present composition. Examples of addable food comprising aforementioned inclusive therein are various food, beverage, gun, vitamin complex, health improving food and the like

[59] Wherein the food additives can be added to one or one over food such as fruits, vegetables, food dehydrated foods or cutting products such as fruits, vegetables; fruit juice, vegetable juices or the mixture juices thereof; drinks containing acid-beverage; confectionaries such as cookie, candy, caramel, gun; breads; ice creams, teas, fermented milk such as yogurt; dairy product, spices, alcoholic beverages, cans, in- bottles, noodles, processed livestock products, processed marine products, fermented food, beans food, cereals food, processed meats, licorices or herbs, specifically, at least one selected from gum, candy, biscuit, ice cream or chocolate, which may be prepared by the methods well-known in the art and shall be explained in detail as follows.

[60]

[61] R>r example, for preparing chewing gun of the present invention, the mixture of 0.5 w/w (%) food additive of the present invention, 25 w/w (%) gum base, and 72 w/w (%) were mixed at pre-heated mixer at 60 to 7(JC and 2.5 w/w (%) peppermint flavor is added thereto to mix together with keeping the internal temperature at 55⁰C and to extract. The resulting base is subject to press, aging, cutting, packaging to produce purposed chewing gum.

[62] The inventive food additive may be used in preparing confectionary or bakery; sauce or dressing material; or fermented milk or yoghurt by adding therein as a form of powder or liquid phase for making functional or preferential food. The inventive food additive may be used in preparing kimchi, fermented salted fish, or salad by directly adding therein for the purpose of the conservation of taste, anti-oxidation, or discoloration. It also may be used in preparing ham, sausage etc. by soaking or spraying method. The powder or granule form of inventive food additive may be used as seasoning by mixing with other spices such as a garlic, a green onion, a ginger, a herb, a licorice root etc., and may be used in processed marine products such as a boiled fish paste, a dried slice of fish seasoned with spices, slated fish or various kimchies such as cabbage kimchi, radish kimchi, etc. by soaking or spraying method.

[63]

[64] The inventive inclusion complex of the present invention also can be used as a main component or additive and aiding agent in the preparation of various functional health food and health care food.

[65] Accordingly, it is the other object of the present invention to provide a functional health food comprising an inventive inclusion complex of the present invention for reinforcing immune resisting power.

[66]

[67] The term " functional health food" defined herein may be the functional food having enhanced functionality such as physical functionality or physiological functionality by adding the inventive inclusion complex of the present invention to conventional food to reinforce immune resisting power in himan or mammal, which comprise various formulations such as tablet, capsule, powder, granule or liquid formulation.

[68]

[69] Inventive composition of the present invention has no toxicity and adverse effect therefore; they can be used with safe.

[70]

[71] It will be apparent to those skilled in the art that various modifications and variations can be made in the compositions, use and preparations of the present invention without departing from the spirit or scope of the invention. [72]

Advantageous Effects

[73] Inventive inclusion complex of present invention could more effectively scavenge the bitter taste by forming the inclusion with bitter components of ginseng extract with holding innate flavor of itself as well as enhance the solubility of ginseng extract, the stability of ginseng extract and the transparency of liquid formulation containing ginseng extract comparing with those of alpha-cyclodextrin and beta-cyclodextrin, being confirmed through various experiments, i.e., a sensory test, transmittance determination test, stability determination test, NMR determination etc.

[74]

Brief Description of the Drawings

[75] The above and other objects, features and other advantages of the present invention will more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which;

[76]

[77] Hg. 1 represents the NMR spectral data of an inventive red ginseng inclusive powder prepared in Example 1-3.

[78]

Best Mode for Carrying Out the Invention

[79] It will be apparent to those skilled in the art that various modifications and variations can be made in the compositions, use and preparations of the present invention without departing from the spirit or scope of the invention.

[80]

Mode for the Invention

[81] The present invention is more specifically explained by the following examples.

However, it should be understood that the present invention is not limited to these examples in any manner.

[82]

[83] Reference Example 1. Preparation of materials

[84] Six years-old cultivated ginseng procured from Keumsan Susan Center (Keumsan

City, Korea) was performed to conventional process for making red ginseng, i.e., repeated steaming steps at 90-95⁰C for 3 hours and drying steps in order that the water amount of ginseng reached to 12.5-13.5 v/w (%), and then subjected to lyophilization to prepare dried red ginseng. Alpha-cyclodextrin (CAVAMAX® W6 Rod), beta- cyclodextrin (CAVAMAX® W7 Rod), and gamma-cyclodextrin (CAVAMAX® W8 Ibod) were procured at commercial manufacturing company (Wacker Chemical Corporation).

[85]

[86]

[87] Example 1. Preparation of the inclusion product of red ginseng using gamma- cyclodextrin

[88] 1-1. Preparation of red ginseng extract

[89] lkg of red ginseng prepared in Reference Example 1 was performed to reflux extraction with 70 v/v(%) ethanol solution, filtration and the filtrate was concentrated with vaccum at 6(JC to the extent that the water amount of solid phase reached to 70 w/v(%) to obtain lkg of red ginseng extract, which was used in following Examples and Comparative Examples.

[90]

[91] 1-2. Preparation of inventive inclusion complex (γ-CDD

[92] lOOg of distilled water was added to lOOg of red ginseng extract prepared in step 1-1 and diluted to the extent that the amount of solid phase reached to 35 w/v(%). 2. Ig of gamma-cyclodextrin (3 w/w% based on the solid phase) was added thereto and the mixture was heated at 85⁰C for 2 hours with stirring to perform inclusive reaction. The resulting product was dried to obtain 68.5g of inventive inclusion complex of red ginseng (designated as "γ-CDl" hereinafter).

[93]

[94] 1-3. Preparation of inventive inclusion complex (γ-CD2)

[95] Excepting that the amount of gamma-cyclodextrin was substituted with 3.5g (5 w/ w(%) based on the solid phase), all the procedure was performed according to the identical procedure disclosed in Step 1-2 to afford 69.8g of inventive inclusion complex of red ginseng (designated as "γ-CD2" hereinafter).

[96]

[97] 1-4. Preparation of inventive inclusion complex (γ-CD3)

[98] Excepting that the amount of gamma-cyclodextrin was substituted with 7g (10 w/ w(%) based on the solid phase), all the procedure was performed according to the identical procedure disclosed in Step 1-2 to afford 73.2g of inventive inclusion complex of red ginseng (designated as "γ-CD3" hereinafter).

[99]

[100] 1-5. Preparation of inventive inclusion complex (γ-CD4) [101] Excepting that the amount of ganrna-cyclodextrin was substituted with 14g (20 w/ w(%) based on the solid phase), all the procedure was performed according to the identical procedure disclosed in Step 1-2 to afford 79.8g of inventive inclusion complex of red ginseng (designated as "γ-CD4" hereinafter).

[102]

[103] 1-6. Preparation of inventive inclusion complex (γ-CD5)

[104] Excepting that the amount of gamma-cyclodextrin was substituted with 2 Ig (30 w/ w(%) based on the solid phase), all the procedure was performed according to the identical procedure disclosed in Step 1-2 to afford 86.5g of inventive inclusion complex of red ginseng (designated as "γ-CD5" hereinafter).

[105]

[106] Comparative Example 1. Preparation of the inclusion complex of red ginseng using alpha-cyclodextrin (α-CDl)

[107] lOOg of distilled water was added to lOOg of red ginseng extract prepared in step 1-1 and diluted to the extent that the amount of solid phase reached to 35 w/v(%). 7g of gamma-cyclodextrin (10 w/w% based on the solid phase) was added thereto and the mixture was heated at 85⁰C for 2 hours with stirring to perform inclusive reaction. The resulting product was performed to spray drying to obtain 73.2g of comparative inclusion complex of red ginseng (designated as "α-CDl" hereinafter).

[108]

[109] Comparative Example 1. Preparation of the inclusion complex of red ginseng using alpha-cyclodextrin (β-CDl)

[110] Excepting the alpha-cyclodextrin was substituted with beta-cyclodextrin, all the procedure was performed according to the identical procedure disclosed in the above Comparative Example 1 to afford 73.1g of comparative inclusion complex of red ginseng (designated as "β-CDl" hereinafter).

[I l l]

[112] Experimental Example 1. Sensory test

[113] To compare the effect of inventive inclusion complexes prepared in Example 1 on the removal of bitter taste with holding innate flavor of red ginseng comparing with those of the comparative inclusion complexes prepared in Comparative Examples, the inclusion complexes were dissolved in distilled water at the concentration of 1 w/w(%) to determine the difference in innate flavor and bitter taste. Hve trained volunteers took participate in the sensory test and the results were scored with the condition that the score of control group treated with none was set to 10 and scores were classified into nine steps pursuant to relative evaluation. Resulting scores were transformed into mean value or mean value standard deviation and the analysis was repeated three times. The significance was verified according to Duncan's multiple range analysis and the result was shown in Table 1.

[114] [115] Table 1 [Table 1] [Table ]

[116] [117] As can be seen in Table 1, the scores to determine bitter taste in the group treated with Comparative Example 1 (α-CDl; about 8.3) and Comparative Example 2 (β-CDl; about 9.0) were higher than that of Example 1-4 with the equivalent amount of cyclodextrin (γ-CD3; about 1.85), which confirms that the bitter taste of inventive inclusion complex prepared in Example 1-4 showed much weaker that those of comparative inclusion complex prepared in Comparative Example 1 and 2. IVbreover, the scores to determine the innate ginseng flavor in the group treated with Comparative Example 1 (α-CDl; about 9.2) and Comparative Example 2 (β-CDl; about 9.3) were higher than that of Example 1-4 with the equivalent amount of cyclodextrin (γ-CD3; about 7.9), which confirms that the innate ginseng flavor of inventive inclusion complex prepared in Example 1-4 showed much stronger that those of comparative inclusion complex prepared in Comparative Examples.

[118] Accordingly, it has been confirmed that the inventive inclusion complex showed potent effect on removing bitter taste with holding its innate flavor.

[119] [120] Experimental Example 2. The determination of transmittance [121] To determine the transmittance of inventive inclusion complexes prepared in Example 1 comparing with those of the comparative inclusion complexes prepared in Comparative Examples, the inclusion complexes were dissolved in distilled water at the concentration of 1 w/w(%) and left alone in incubator at 45⁰C for 72 hours to observe the precipitates. The transmittance was determined by UV/vis spectrophotometer (V-550, JASCO) at 600nm and the result was shown in Table 2.

[122] [123] Table 2 [Table 2] [Table ]

[124] [125] As can be seen in Table 2, the turbidity of the group treated with γ-CD2 (5 w/v%) and γ-CD3 (10 w/v%) was very lower than that the control group. However the turbidity of the group treated with more than those concentrations got worsen reversely, which means that the solubility in case treatment group with over dose of gamma-cyclodextrin becomes reduced although the gamma-dextrin itself provides enhanced solubility. Accordingly, it has been confirmed that the concentrations of gamma-cyclodextrin of 5 or 10 w/v% are optimum to prepare liquid formulation.

[126] [127] Experimental Example 3. The determination of stability [128] To determine the stability of inventive inclusion complexes prepared in Example 1 comparing with those of the comparative inclusion complexes prepared in Comparative Examples, following test was performed as follows.

[129] Ginsenoside RbI, an active ingredient of inventive inclusion complexes (γ-CD2), was prepared by following procedure:

[130] Ig of γ-CD2 prepared in Example 1-3 was poured in round flask and 70 ml of 50% (v/v) methanol was added thereto to perform reflux extraction with heating in water bath for 1 hour. The solution was centrifuged to collect supernatant and the remaining residue was performed to repeated extraction with 50 ml of 50% (v/v) methanol to collect supernatant. The collected supernatants were mixed and concentrated with vacuun at 6OC. 20ml of buffer solution was added thereto and the solution was transferred to 25ml volune of flask. The volune of flask was adjusted to 25ml by adding small quantity of buffer solution. 5ml of solution was run into activated C18 cartridge colunn (Sep-Pak Plus, WAT020515, Waters, USA) and the column was washed with water and 30% (v/v) methanol to elute with 20ml of methanol. The eluted solution was concentrated with vacuun below 6OC and dissolved in 2ml of methanol to be used in following analysis.

[131] The amount of ginsenoside RbI, an active ingredient of inventive inclusion complex (γ-CD2), was determined by using HPLC (717 plus autosampler, 1525 binary pump and 2487 dual wavelength UV detector, Waters, US) equipped with colunn (Cap-Call Pak C 18, 250x4.6 mm, 5 micrometer, Shiseido, JAPAN) at 203nm. The mixture solution (CH3CN:water=30:70) was used as a mobile phase running at the speed of 1.5 ml/min. various concentrations of standard sample solution, i.e., 1, 0.1, 0.01 mg/ml were prepared from stock solution prepared by dissolving ginsenoside RbI (Sigma, St- Louis, MD, USA) in methanol to the concentration of lmg/ml to obtain calibration curve. The change in the amount of ginsenoside RbI in γ-CD2 was determined and the result was shown in Table 3

[132]

[133] Table 3 [Table 3] [Table ]

[134] [135] Aa can be seen in Table 3, the amount of ginsenoside RbI in γ-CD2 has sustained to about 97% after three months, which confirms that the inventive inclusion complex is proved to be stable regardless of the change of active ingredient.

[136] [137] Experimental Example 4. The determination of NMR [138] To determine whether the ginseng extract formed inclusion complex with gamma- cyclodextrin or not, the structure of γ-CD5 prepared in Example 1-6 was analyzed with iH-NMR spectran (Varian UNITY INOVA 400; 400MHz) adopting the chemical shift value of water peak as an internal standard. Gamma-cyclodextrin and γ-CD5 prepared in Example 1-6 were dissolved in D₂O (Sigma, St-Louis, MO, USA) to determine and the result was shown in Hg. 1.

[139] [140] At the result, the internal pore widths of gamma-cyclodextrin, i.e., H3 (3.965) and H6 (3.901) showed chemical shits to that of γ-CD5, i.e., H3 (3.965) and H6 (3.901), which showed similar tendency to the result of an inclusive formation with gamma- cylodextrin and ferulic acid (J. Pharm. Biomed. Anal., 40, pp875-881, 2006). Accordingly, it has been confirmed that ginseng extract definitely formed inclusion complex with gamma-cyclodextrin being proved by the chemical- shift in proton atom positioned at internal pore and therefore the inventive inclusion complex could effectively scavenge the bitter taste by forming the inclusion with bitter components of ginseng extract with holding innate flavor of itself as well as enhance the solubility of ginseng extract, the stability of ginseng extract and the transmittance of liquid formulation containing ginseng extract comparing with those of alpha-cyclodextrin and beta-cyclodextrin.

[141] [142] Hereinafter, the formulating methods and kinds of food compositions or health functional food will be described, but the present invention is not limited to them.

[143] Preparation of chewing gun(A)

[144] γ-CDl 0.5 w/w%

[145] Gun base 20 w/w%

[146] Sugar 76.5 w/w%

[147] Fruit Flavor 1.0 w/w%

[148] Distilled water 2.0 w/w%

[149]

[150] Chewing gun was prepared by mixing the above components and by the method well-known in the art.

[151]

[152] Preparation of chewing gun (B)

[153] γ-CD2 0.5 w/w%

[154] Gun base 25 w/w%

[155] Sorbitol 72.0 w/w%

[156] Peppermint Flavor 2.5 w/w%

[157] Distilled water optimum amount

[158]

[159] Chewing gun was prepared by mixing the above components and by the method well-known in the art.

[160]

[161] Preparation of candv(A)

[162] γ-CD3 0.4 w/w%

[163] Sugar 54.5 w/w%

[164] Starch syrup 45.0 w/w%

[ 165] Orange Flavor 0.1 w/w%

[166]

[167] Candy was prepared by mixing the above components and by the method well- known in the art.

[168]

[169]

[170] Preparation of candy (B)

[171] γ-CD4 0.5w/w%

[172] Malditol 54.5 w/w% [173] Reduced Starch syrup 45.0 w/w%

[174] Natural Herb Flavor 0.1 w/w%

[175]

[176] Candy was prepared by mixing the above components and by the method well- known in the art.

[177]

[178] Preparation of biscuit

[179] γ-CD5 1.7kg

[180] Weak flour (1^st grade) 88kg

[181] Medium flour ( 1 ^st grade) 76.4kg

[182] Refined sugar 16.5kg

[183] Table salt 2.5kg

[184] Glucose 2.7kg

[ 185] Palm shortening 40.5kg

[186] Sodiun hydrogen carbonate 0.6kg

[187] Sodiun bisulfite 0.55kg

[188] Rice powder 5.0kg

[189] Vitamin Bl 0.003kg

[190] Vitamin B2 0.003kg

[191] MIk flavor 0.16kg

[192] Distilled water 71. lkg

[193] Whole milk powder 4.0kg

[194] Substitute milk powder 1.Okg

[195] Calcium phosphate monobasic 0. lkg

[196] Scorched salt 1.0kg

[197] Spray milk 25 kg

[198]

[199] Biscuit was prepared by mixing the above components and by the method well- known in the art.

[200]

[201]

[202] Preparation of ice cream

[203] γ-CDl 0.5 w/w%

[204] Fatty oil 10.0 w/w%

[205] Solid-not-fat 10.0 w/w% [206] Sugar 12.0 w/w%

[207] Starch syrup 3.0 w/w%

[208] Emulsifier (Span) 0.5 w/w%

[209] Flavor (Strawberry) 0.15 w/w%

[210] Distilled water 63.05 w/w%

[211]

[212] Ice cream was prepared by mixing the above components and by the method well- known in the art.

[213]

[214] Preparation of chocolat

[215] γ-CD2 0.5 w/w%

[216] Cocoa butter 34.0 w/w%

[217] Cocoa mass 15.0 w/w%

[218] Cocoa powder 15.0 w/w%

[219] Lecithin 0.5 w/w%

[220] Flavor (Vanilla) 0.5 w/w%

[221]

[222] Chocolate was prepared by mixing the above components and by the method well- known in the art.

[223]

[224] Preparation of table

[225] γ-CD3 200 mg

[226] Corn Starch 100 mg

[227] Lactose 100 mg

[228] Magnesium Stearate optimum

[229]

[230] Tablet preparation was prepared by mixing above components and entabletting.

[231]

[232] Preparation of capsule

[233] γ-CD4 100 mg

[234] Corn Starch 50 mg

[235] Lactose 50 mg

[236] Talc 2 mg

[237] Lactose 14.8 mg

[238] Magnesium Stearate optimum [239]

[240] Capsule preparation was prepared by mixing above components and filling gelatin capsule by conventional gelatin preparation method.

[241]

[242] Preparation of liquid

[243] γ-CD5 1000 mg

[244] Sugar 20 g

[245] Isomerized sugar 20 g

[246] Lemon flavor optimum amount

[247]

[248] Liquid formulation was prepared by dissolving the components to distilled water with a proper dose of lemon scent, mixing, adjusting to 100 ml with distilled water in brown bottle and sterilizing by conventional liquid formulation preparation method.

[249]

[250] Preparation of health functional food

[251] γ-CDl 1000 mg

[252] Vitamin mixture optimum amount

[253] Vitamin A acetate 70 μg

[254] Vitamin E 1.0 mg

[255] Vitamin B ₁ 0.13 mg

[256] Vitamin B ₂0.15 mg

[257] Vitamin B ₆0.5 mg

[258] Vitamin B ₁₂ 0.2 μg

[259] Vitamin C 10 mg

[260] Biotin 10 μg

[261] Amide nicotinic acid 1.7 mg

[262] E)lic acid 50 μg

[263] Calcium pantothenic acid 0.5 mg

[264] Mineral mixture optimum amount

[265] Ferrous sulfate 1.75 mg

[266] Zinc oxide 0.82 mg

[267] Magnesium carbonate 25.3 mg

[268] Monopotassium phosphate 15 mg

[269] Dicalcium phosphate 55 mg

[270] Potassium citrate 100 mg [271 ] Magnesium chloride 24.8 mg

[272]

[273] The above -mentioned vitamin and mineral mixture may be varied in many ways.

Such variations are not to be regarded as a departure from the spirit and scope of the present invention.

[274]

[275] Preparation of health beverage

[276] γ-CD2 1000 mg

[277] Citric acid 1000 mg

[278] Oligosaccharide 100 g

[279] Apricot concentration 2 g

[280] Taurine 1 g

[281] Distilled water 900 ml

[282]

[283] Health beverage preparation was prepared by dissolving active component, mixing, stirring at 85⁰C for 1 hour, filtering and then filling all the components in 2000 ml container and sterilizing by conventional health beverage preparation method.

[284]

[285] The invention being thus described may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

[286]

Industrial Applicability

[287] As described in the present invention, the inventive inclusion complex could effectively scavenge the bitter taste by forming the inclusive with bitter components of ginseng extract with holding innate flavor itself as well as enhance the solubility of ginseng extract, the stability of ginseng extract and the transmittance of liquid formulation containing ginseng extract. Therefore, it can be useful in the preparation of health food, food additive and health functional food with good quality.

Claims

[1] A method for preparing the inclusion complex of ginseng extract with garrma- cyclodextrin comprising the steps consisting of: extracting a ginseng material with 1 to 20-fold volume of water, ethanol or the mixture thereof and concentrating the filtrate with filtering the residue to obtain ginseng extract at the 1^st step; adding distilled water to the extract to obtain diluted solution at the 2^nd step; and adding garrma-cyclodextrin thereto with stirring to perform inclusive reaction at the 3^rd step.

[2] The method according to claim 1, wherein said method comprises the steps consisting of: extracting a ginseng material with 3 to 10-fold volume of water, ethanol or the mixture thereof for the period ranging from 1 hour to 10 hours once to ten times at the temperature ranging from 70 to 100 ⁰C with a hot- water extraction, ultrasonication extraction or reflux extraction filtering the solution to obtain filtrate and concentrating the filtrate with vacuum at the temperature ranging from 40 to 80 ⁰CtO obtain ginseng extract at the 1 ^st step; adding distilled water to the extract to the extent that the amount of solid phase reaches to the range from 10 to 80 w/v% to obtain diluted solution at the 2^nd step; and adding 0.1 to 99.9 w/w% garrma-cyclodextrin based on the amount of solid phase thereto with stirring at the temperature ranging from 40 to 120 ⁰C for the period ranging from 0.5 hour to 6 hours to perform inclusive reaction at the 3 ^rd step.

[3] The method according to claim 1, wherein said ginseng material comprises

Panax ginseng, Panax quinquefolia, Panax notoginseng, Panax japonica, Panax trifolia, Panax pseudoginseng, Panax vietnamensis, Panax elegatior, Panax wangianus or Panax bipinratifidus in the form of a red ginseng, white ginseng, black ginseng, unhusked ginseng, husked ginseng, wild ginseng or cultivated ginseng

[4] A food or food additive composition comprising the inventive inclusion complex prepared by the method as set forth in claim 1.

[5] The food or food additive composition according to claim 4, wherein said composition comprises confectionary such as chewing gum, candy, biscuits, chocolate and ice cream.

[6] A health functional food comprising the inventive inclusion complex prepared by the method as set forth in claim 1.

[7] The health functional food according to claim 6, wherein said food comprises the form of tablet, capsule, powder, granule or liquid formulation.