KR100612377B1 - Composition comprising hot water extract of Phellinus mushroom fruit body for preventing and treating acute resparatory disease - Google Patents

Composition comprising hot water extract of Phellinus mushroom fruit body for preventing and treating acute resparatory disease Download PDF

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KR100612377B1
KR100612377B1 KR1020060039900A KR20060039900A KR100612377B1 KR 100612377 B1 KR100612377 B1 KR 100612377B1 KR 1020060039900 A KR1020060039900 A KR 1020060039900A KR 20060039900 A KR20060039900 A KR 20060039900A KR 100612377 B1 KR100612377 B1 KR 100612377B1
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박승춘
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경북대학교 산학협력단
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Abstract

본 발명은 진흙버섯 자실체의 열수 추출물을 유효성분으로 하는 급성 호흡기 질환의 예방 및 치료용 조성물에 관한 것이다.The present invention relates to a composition for the prevention and treatment of acute respiratory diseases comprising the hot water extract of the fruiting mud mushroom body.

본 발명의 진흙버섯 자실체의 열수 추출물은 백혈구의 수를 감소시키고, 사이토킨의 농도를 현저히 감소시킴으로, 급성 호흡기 염증 억제 효과가 우수하다.The hydrothermal extract of the mud mushroom fruiting body of the present invention reduces the number of white blood cells and significantly reduces the concentration of cytokines, and is excellent in inhibiting acute respiratory inflammation.

따라서, 본 발명의 조성물은 감기, 급성 인두염, 급성 비염, 급성 부비강염, 급성 편도염, 급성 후두염, 급성 후두개염, 급성 기관지염, 급성 상기도 감염증 및 급성 호흡 궁박증후군과 같은 급성 호흡기 질환의 예방 및 치료에 유용하게 사용될 수 있다.Therefore, the composition of the present invention is for the prevention and treatment of acute respiratory diseases such as cold, acute pharyngitis, acute rhinitis, acute sinusitis, acute tonsillitis, acute laryngitis, acute laryngitis, acute bronchitis, acute upper respiratory infection and acute respiratory distress syndrome. It can be usefully used.

Description

진흙버섯 자실체의 열수 추출물을 유효성분으로 하는 급성 호흡기 질환의 예방 및 치료용 조성물{Composition comprising hot water extract of Phellinus mushroom fruit body for preventing and treating acute resparatory disease}Composition comprising hot water extract of Phellinus mushroom fruit body for preventing and treating acute resparatory disease}

도 1은 본 발명의 진흙버섯 자실체 열수 추출물의 항암활성에 대한 반응표면분석(a) 및 등고선도(b)를 나타낸 도이다.1 is a view showing the response surface analysis (a) and contour diagram (b) for the anticancer activity of the mud mushroom fruiting body hydrothermal extract of the present invention.

도 2는 본 발명의 진흙버섯 자실체 열수 추출물의 총당에 대한 반응표면분석 (a) 및 등고선도(b)를 나타낸 도이다.Figure 2 is a view showing the response surface analysis (a) and contour diagram (b) of the total sugar of the mud mushroom fruiting body hydrothermal extract of the present invention.

도 3은 본 발명의 진흙버섯 자실체 열수 추출물의 환원당에 대한 반응표면분석(a) 및 등고선도(b)를 나타낸 도이다.Figure 3 is a view showing the reaction surface analysis (a) and contour diagram (b) for the reducing sugar of the mud mushroom fruiting body hydrothermal extract of the present invention.

본 발명은 진흙버섯 자실체의 열수 추출물을 유효성분으로 하는 급성 호흡기 질환의 예방 및 치료용 조성물에 관한 것이다.The present invention relates to a composition for the prevention and treatment of acute respiratory diseases comprising the hot water extract of the fruiting mud mushroom body.

현대인들은 산업의 발달로 대기오염, 알레르기원(동물의 털, 꽃가루 등), 흡 연 등으로 인한 알레르기성 기관지 질환에 노출되어 있으며, 감기, 후두염, 급성 후두개염, 세기관지염, 폐렴, 천식, 편도선염 등과 같은 호흡기 질환은 선진국에서 가장 빈번하게 발생하는 질환 중 하나로 성인보다는 어린아이에게서 많이 발생된다는 보고가 있다(Sears, 1997).Modern people are exposed to allergic bronchial diseases caused by air pollution, allergens (animal hair, pollen, etc.), smoking, etc. due to the development of industry, cold, laryngitis, acute laryngitis, bronchiolitis, pneumonia, asthma, tonsillitis, etc. The same respiratory disease is one of the most frequent diseases in developed countries and is reported to occur more frequently in children than in adults (Sears, 1997).

매년 5세 이하의 1,500만명의 어린이 중 약 400만명이 급성 호흡기질환 (acute resparatory disease)으로 사망하고 있으며, 이 사망자 중 90% 이상이 개발도상국가에서 일어난다. 또한, 이러한 호흡기 질환은 특히 대기 환경 오염이 심각한 지역에서 발생빈도가 높다.Each year, about 4 million of the 15 million children under age 5 die from acute resparatory disease, and more than 90% of these deaths occur in developing countries. In addition, such respiratory diseases are frequently occurring in areas where air pollution is serious.

예로부터 버섯은 항암, 항산화, 콜레스테롤 저하, 혈압강하, 면역강화, 종양저지, 자궁출혈, 월경불순 등의 치료에 효과가 좋은 것으로 알려져 왔다.Mushrooms have been known to be effective in the treatment of anticancer, antioxidant, cholesterol lowering, blood pressure lowering, immune strengthening, tumor suppression, uterine bleeding, and menstrual irregularities.

최근에는 이들 버섯 중 진흙버섯류의 추출물이 건강보조식품으로 각광을 받고있다. 진흙버섯은 민주름버섯목(Aphylophorales), 소나무비닐버섯과 (Hymenochaetaceae), 진흙버섯(Phellinus)에 속하는 백색 부후균으로 주로 참나무, 뽕나무와 같은 활엽수에 발생하는데, 표고버섯 재배시 참나무 골목에 발생하여 병을 일으킨다(차 등, 1994). 자실체는 코르크질로 단년생 또는 다년생으로 알려져 있으며, 나무에 부착된 편반구형으로 보통 상하가 서로 연결되어 있는 군생형으로 발생하며 크기는 지름 5~12㎝ 정도로 상층표면은 암황갈색으로 융모가 있고 가장 자리는 갓에 비해서 색이 엷고 관공의 표면은 암자갈색으로 ㎜당 6~8개가 있으며 포자는 4~5 x 3~3.5 ㎛로 타원형이며 표면은 평활하다고 알려져 있다(Ryvardan, 1993).Recently, the extract of mud mushrooms among these mushrooms has been spotlighted as a dietary supplement. Mud mushrooms are white fungi belonging to Aphylophorales, Hemenochaetaceae and Phellinus. They occur mainly on hardwoods such as oak and mulberry. Cause illness (tea et al., 1994). The fruiting body is cork, known as short-term or perennial. It is a hemispherical shape attached to the tree and usually occurs as a colony type with upper and lower sides connected to each other. The upper surface is dark yellow brown with villi and edges. Are lighter in color than the lampshade, and the surface of the cavities is dark brown, 6-8 per mm, spores are 4-5 x 3 ~ 3.5 ㎛, elliptical, and the surface is known to be smooth (Ryvardan, 1993).

이에, 본 발명자들은 반응표면분석법에 의한 최적의 추출조건으로 진흙버섯 자실체로부터 얻은 열수 추출물에서 급성 호흡기 염증을 억제하는 효과가 있음을 발견하고 본 발명을 완성하였다.Thus, the present inventors have found that the optimum extraction conditions by the reaction surface analysis method has the effect of inhibiting acute respiratory inflammation in the hydrothermal extract obtained from the mud mushroom fruiting body and completed the present invention.

본 발명은 진흙버섯 자실체의 열수 추출물을 유효성분으로 하는 급성 호흡기 질환의 예방 및 치료용 조성물을 제공하고자 한다.The present invention is to provide a composition for the prevention and treatment of acute respiratory diseases, the active ingredient of the hot water extract of the mud mushroom fruiting body.

본 발명은 진흙버섯 자실체의 열수 추출물을 유효성분으로 하는 급성 호흡기 질환의 예방 및 치료용 조성물을 제공한다.The present invention provides a composition for the prevention and treatment of acute respiratory diseases comprising the hot water extract of the mud mushroom fruiting body as an active ingredient.

상기 진흙버섯 자실체의 열수 추출물은 반응표면분석법을 이용하여 추출조건을 최적화하여 추출함을 특징으로 한다.The hydrothermal extract of the mud mushroom fruiting body is characterized by extracting by optimizing the extraction conditions using the reaction surface analysis method.

이하, 본 발명에 대해 상세히 설명한다.Hereinafter, the present invention will be described in detail.

본 발명에서 사용되는 진흙버섯 자실체는 참나무 톱밥을 121℃에서 90분간 고압살균하여 참나무 톱밥배지를 만든 후 버섯균을 접종하여 3개월 만에 속성으로 재배한 것을 사용한다.The mud mushroom fruiting body used in the present invention is used by autoclaving oak sawdust at 121 ° C. for 90 minutes to make oak sawdust medium, and then inoculating mushroom bacteria and cultivating it as a property in three months.

본 발명에서 사용되는 진흙버섯은 펠리너스 린테우스(Phellinus linteus), 펠리너스 바우미(Phellinus baumii), 펠리너스 길버스(Phellinus gilvus) 중에서 선택된 것을 사용한다.The mud mushroom used in the present invention uses one selected from Pelinus linteus, Pelinus baumii, and Pelinus gilvus.

본 발명의 진흙버섯 자실체의 열수 추출물은 중심합성계획법에 의한 반응표면분석법을 이용하여 다음과 같은 방법으로 추출조건을 최적화하여 추출한다.The hot water extract of the mud mushroom fruiting body of the present invention is extracted by optimizing the extraction conditions by the following method using the reaction surface analysis method according to the central synthesis planning method.

중심합성계획에 의한 독립변수(Xi)는 추출시간(X1; 8, 10, 12시간), 추출온도(X2; 80, 90, 100℃) 및 추출부피(X3; 10, 20, 30㎖/g)로 구성하며, 이들 독립변수에 의해 영향을 받는 종속변수(Yn)는 항암활성(Y1), 총당(Y2) 및 환원당(Y3)으로 하고, 이들은 3회 반복 측정하여 그 평균값을 회귀분석에 사용한다.Independent variables (X i ) according to the central synthesis plan were extracted time (X 1 ; 8, 10, 12 hours), extraction temperature (X 2 ; 80, 90, 100 ° C) and extraction volume (X 3 ; 10, 20, 30 ml / g), and the dependent variable (Y n ) affected by these independent variables is anticancer activity (Y 1 ), total sugar (Y 2 ) and reducing sugar (Y 3 ), which are repeated three times. The mean value is used for the regression analysis.

본 발명의 진흙버섯 자실체의 열수 추출물의 항암활성의 회귀계수는 0.9578 이며, 추출온도와 추출부피에 비례한다. 항암활성에 대한 진흙버섯 자실체의 열수 추출물의 최적 조건은 추출시간 10.24시간, 추출온도 99.46℃, 추출부피 23.00㎖/g 이며, 85.5%에서 최대반응이 나타난다.The regression coefficient of anticancer activity of the hydrothermal extract of the mud mushroom fruiting body of the present invention is 0.9578, which is proportional to the extraction temperature and the extraction volume. The optimum conditions of hydrothermal extract of mud mushroom fruiting body against anticancer activity were extraction time of 10.24 hours, extraction temperature of 99.46 ℃, extraction volume of 23.00ml / g, and maximum response at 85.5%.

본 발명의 진흙버섯 자실체의 열수 추출물의 총당의 회귀계수는 0.9276 이며, 추출온도에만 비례한다. 총당에 대한 진흙버섯 자실체의 열수 추출물의 최적 조건은 추출시간 10.70시간, 추출온도 99.13℃, 추출부피 17.90㎖/g 이며, 2.08㎎/㎖의 농도에서 최대반응이 나타난다.The regression coefficient of the total sugar of the hydrothermal extract of the mud mushroom fruiting body of the present invention is 0.9276, which is proportional only to the extraction temperature. The optimum conditions of hydrothermal extract of mud mushroom fruiting body for total sugar were extraction time of 10.70 hours, extraction temperature of 99.13 ℃, extraction volume of 17.90ml / g, and maximum reaction at the concentration of 2.08mg / ml.

본 발명의 진흙버섯 자실체의 열수 추출물의 환원당의 회귀계수는 0.9832 이며, 추출온도와 추출부피에 비례한다. 환원당에 대한 진흙버섯 자실체의 열수 추출의 최적 조건은 추출시간 10.27시간, 추출온도 96.11℃, 추출부피 27.80㎖/g 이며, 890.6ppm에서 최대반응이 나타난다.The regression coefficient of the reducing sugar of the hydrothermal extract of the mud mushroom fruiting body of the present invention is 0.9832, which is proportional to the extraction temperature and the extraction volume. The optimum conditions for hot water extraction of mud mushroom fruiting bodies for reducing sugars were extraction time of 10.27 hours, extraction temperature of 96.11 ℃, extraction volume of 27.80ml / g, and maximum reaction at 890.6ppm.

따라서, 본 발명의 진흙버섯 자실체의 열수 추출물의 최적의 추출조건은 추출시간 10~11시간, 추출온도 96~100℃, 추출부피 17~28㎖/g 로 결정할 수 있다.Therefore, the optimum extraction conditions of the hydrothermal extract of the mud mushroom fruiting body of the present invention can be determined by the extraction time 10 ~ 11 hours, the extraction temperature 96 ~ 100 ℃, the extraction volume 17 ~ 28mL / g.

본 발명의 진흙버섯 자실체의 열수 추출물은 백혈구의 수를 감소시키고, 사이토킨의 농도를 현저히 감소시킴으로, 감기, 급성 인두염, 급성 비염, 급성 부비강염, 급성 편도염, 급성 후두염, 급성 후두개염, 급성 기관지염, 급성 상기도 감염증 및 급성 호흡 궁박증후군과 같은 급성 호흡기 질환의 예방 및 치료에 유용하게 사용될 수 있다.The hot water extract of the mud mushroom fruiting body of the present invention reduces the number of leukocytes and significantly reduces the concentration of cytokines, such as cold, acute pharyngitis, acute rhinitis, acute sinusitis, acute tonsillitis, acute laryngitis, acute laryngitis, acute bronchitis, and acute It can be usefully used for the prevention and treatment of acute respiratory diseases such as upper respiratory tract infection and acute respiratory distress syndrome.

본 발명의 조성물은 진흙버섯 자실체의 열수 추출물에 추가로 동일 또는 유사한 기능을 나타내는 유효성분을 1종 이상 함유할 수 있다.The composition of the present invention may contain one or more active ingredients exhibiting the same or similar function in addition to the hydrothermal extract of the mud mushroom fruiting body.

본 발명의 조성물은, 투여를 위해서 상기 기재한 유효성분 이외에 추가로 약제학적 또는 식품에서 허용 가능한 담체를 1종 이상 포함할 수 있다. 약제학적 또는 식품에서 허용 가능한 담체는 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다. 더 나아가 당분야의 적정한 방법으로 또는 Remington's Pharmaceutical Science(최근판), Mack Publishing Company, Easton PA에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화 할 수 있다.The composition of the present invention, in addition to the above-described active ingredient for administration may further comprise one or more carriers which are acceptable in pharmaceutical or food. Pharmaceutical or food acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, as necessary. And other conventional additives such as buffers and bacteriostatic agents can be added. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

본 발명의 조성물은 목적하는 방법에 따라 비경구 투여하거나 경구 투여할 수 있으며, 경구투여가 바람직하다. 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도 등에 따라 그 범위가 다양하나, 일일 투여량은 0.01 ~ 5 g/㎏의 양을 일회 내지 수회에 나누어 투여하는 것이 바람직하다.The composition of the present invention can be administered parenterally or orally according to the desired method, oral administration is preferred. The dosage varies depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, excretion rate and severity of the disease, but the daily dosage is 0.01 to 5 g / kg. It is preferable to administer dividedly to several times.

본 발명의 조성물은 급성 호흡기 질환의 예방 및 치료를 위하여 단독으로, 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 및 생물학적 반응 조절제를 사용하는 방법들과 병용하여 사용할 수 있다.The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of acute respiratory diseases.

본 발명의 조성물은 급성 호흡기 질환의 개선을 목적으로 건강식품에 첨가될 수 있다. 본 발명의 진흙버섯 자실체의 열수 추출물을 식품 첨가물로 사용할 경우, 상기 진흙버섯 자실체의 열수 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에는 본 발명의 진흙버섯 자실체의 열수 추출물은 원료에 대하여 1~20 중량%, 바람직하게는 5~10 중량%의 양으로 첨가된다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.The composition of the present invention can be added to health food for the purpose of improving acute respiratory disease. When the hot water extract of the mud mushroom fruiting body of the present invention is used as a food additive, the hot water extract of the mud mushroom fruiting body may be added as it is, or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, the hot water extract of the mud mushroom fruiting body of the present invention is added in an amount of 1 to 20% by weight, preferably 5 to 10% by weight, based on the raw materials. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. .

상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식 품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of food. Examples of foods to which the above substances can be added include dairy products including meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

본 발명의 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100㎖당 일반적으로 약 0.01~0.04g, 바람직하게는 약 0.02~0.03g 이다.The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

상기 외에 본 발명의 조성물은 여러가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.01~0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonic acid. Carbonating agents and the like used in beverages. In addition, the composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예 및 실험예를 제시한다. 그러나 하기의 실시예 및 실험예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited by the Examples.

실시예Example : 중심합성계획법에 의한 반응표면분석법을 이용한 진흙버섯 자실체의 열수 추출물의 제조 : Preparation of Hot Water Extract of Mud Mushroom Fruits by Response Surface Methodology by Central Synthesis Scheme

본 발명에서 사용한 진흙버섯(Phellinus baumii)은 경상북도 농업기술원에서 속성으로 재배한 자실체를 공급받아 사용하였다.Mud mushroom ( Phellinus baumii ) used in the present invention was used to receive the fruiting body cultivated as a property at Gyeongsangbuk-do Agricultural Research and Extension Services .

1. 중심합성계획법에 의한 반응표면분석법을 이용한 진흙버섯 자실체의 열수1.Heat of Mud Mushroom Fruit Body Using Response Surface Methodology by Central Synthesis Planning 추출 조건의 최적화Optimization of Extraction Conditions

본 발명의 진흙버섯 자실체의 열수 추출물은 반응표면분석법(response surface methodology, RSM)을 이용하여 최적의 추출조건의 예측과 추출물의 이화학적 특성을 모니터링하였다.The hydrothermal extract of the mud mushroom fruiting body of the present invention predicted the optimum extraction condition and monitored the physicochemical properties of the extract by using response surface methodology (RSM).

추출조건의 최적화를 위한 실험계획은 중심합성계획법(central composite design)에 의하여 설계하였고, 반응표면분석 회귀분석을 위해서는 SAS(statistical analysis system) 프로그램을 사용하였다.The experimental design for the optimization of extraction condition was designed by central composite design, and SAS (statistical analysis system) program was used for response surface analysis regression analysis.

중심합성계획에 의한 독립변수(Xi)의 실험계획은 추출공정에서 중요한 변수로 고려되는 인자, 즉 추출시간(X1), 추출온도(X2) 및 추출부피(X3)를 -1, 0, 1의 3 단계로 부호화하여 중심합성계획에 따라 16구로 설정하여 추출실험을 실시하였다. 추출시간(X1)은 8, 10, 12시간, 추출온도(X2)는 80, 90, 100℃ 및 추출부피(X3)는 10, 20, 30㎖/g 으로 구성하였다.The experimental design of the independent variable (X i ) by the central synthesis plan is based on the factors considered as important variables in the extraction process: extraction time (X 1 ), extraction temperature (X 2 ) and extraction volume (X 3 ). Extraction experiments were carried out by encoding 16 steps according to the central synthesis plan by encoding 0 and 1 steps. Extraction time (X 1 ) was composed of 8, 10, 12 hours, extraction temperature (X 2 ) is 80, 90, 100 ℃ and extraction volume (X 3 ) was composed of 10, 20, 30ml / g.

또한 이들 요인변수에 의해 영향을 받는 종속변수(Yn)는 추출물의 품질인자로서 항암활성(Y1), 총당(Y2) 및 환원당(Y3)으로 하였으며, 이들은 3회 반복 측정하여 그 평균값을 회귀분석에 사용하였다.In addition, the dependent variable (Y n ) influenced by these factor variables was the anticancer activity (Y 1 ), total sugar (Y 2 ) and reducing sugar (Y 3 ) as the quality factors of the extracts. Was used in the regression analysis.

진흙버섯 자실체의 열수추출을 최적화하기 위한 중심합성계획은 표 1에 나타내었다.The central synthesis plan for optimizing the hydrothermal extraction of mud mushroom fruiting bodies is shown in Table 1.

진흙버섯 자실체의 열수추출을 최적화 하기 위한 중심합성계획Central Synthesis Scheme to Optimize Hot Water Extraction of Mud Mushroom Fruits 추출시간(X1) (hr)Extraction time (X 1 ) (hr) 추출온도(X2) (℃)Extraction temperature (X 2 ) (℃) 추출부피(X3) (㎖/g)Extraction volume (X 3 ) (ml / g) 1One 8(-1)8 (-1) 80(-1)80 (-1) 10(-1)10 (-1) 22 12(1)12 (1) 80(-1)80 (-1) 10(-1)10 (-1) 33 8(-1)8 (-1) 100(1)100 (1) 10(-1)10 (-1) 44 12(1)12 (1) 100(1)100 (1) 10(-1)10 (-1) 55 8(-1)8 (-1) 80(-1)80 (-1) 30(1)30 (1) 66 12(1)12 (1) 80(-1)80 (-1) 30(1)30 (1) 77 8(-1)8 (-1) 100(1)100 (1) 30(1)30 (1) 88 12(1)12 (1) 100(1)100 (1) 30(1)30 (1) 99 8(-1)8 (-1) 90(0)90 (0) 20(0)20 (0) 1010 12(1)12 (1) 90(0)90 (0) 20(0)20 (0) 1111 10(0)10 (0) 80(-1)80 (-1) 20(0)20 (0) 1212 10(0)10 (0) 100(1)100 (1) 20(0)20 (0) 1313 10(0)10 (0) 90(0)90 (0) 10(-1)10 (-1) 1414 10(0)10 (0) 90(0)90 (0) 30(1)30 (1) 1515 10(0)10 (0) 90(0)90 (0) 20(0)20 (0) 1616 10(0)10 (0) 90(0)90 (0) 20(0)20 (0)

2. 진흙버섯 자실체의 열수 추출물의 제조2. Preparation of hydrothermal extract of mud mushroom fruiting body

진흙버섯의 자실체를 1~2㎝의 크기로 절편한 다음, 흐르는 물에 세척하여 불순물을 제거하였다. 절편된 진흙버섯 자실체는 중심합성계획에 따른 조건에 따라 하기와 같은 열수 추출 순환법으로 추출하였다.The fruiting bodies of the mud mushrooms were cut to a size of 1 to 2 cm, and then washed with running water to remove impurities. The sliced mud mushroom fruiting body was extracted by the following hydrothermal extraction circulation method according to the conditions according to the central synthesis plan.

절편된 진흙버섯 자실체 100g에 2ℓ의 증류수를 첨가하여(진흙버섯 자실체 : 증류수 = 1 : 20), 열수 추출 순환법으로 추출하였다.2 liters of distilled water was added (mud mushroom fruiting body: distilled water = 1:20) to 100 g of the sliced mud mushroom fruiting body, and extracted by hot water extraction circulation method.

열수 추출 순환법은 증류수가 2ℓ에서 0.5ℓ로 증류되면 다시 순환하여 2ℓ가 되게 하는 방법으로, 추출조건에 따라 총 8시간 동안 90℃에서 열수 추출하였다.In the hot water extraction circulation method, when distilled water is distilled from 2 L to 0.5 L, it is circulated again to be 2 L, and hot water is extracted at 90 ° C. for a total of 8 hours depending on extraction conditions.

상기 얻어진 열수 추출액은 1500 xg, 20분동안 원심분리하여 상층액을 얻었다. 얻어진 상층액을 60℃에서 감압농축한 후, 동결건조하여 진흙버섯 자실체의 열수 추출물을 얻었다.The obtained hot water extract was centrifuged at 1500 xg for 20 minutes to obtain a supernatant. The obtained supernatant was concentrated under reduced pressure at 60 ° C., and then lyophilized to obtain a hot water extract of the fruiting mud mushroom.

실험예 1Experimental Example 1 : 진흙버섯 자실체의 열수 추출물의 항암활성, 총당 및 환원당 측정 : Anticancer Activity, Total Sugar and Reducing Sugar of Hydrothermal Extract of Fruits from Mud Mushrooms

본 발명의 진흙버섯 자실체의 열수 추출물의 성분변화를 알아보기 위하여, 항암활성, 총당 및 환원당의 함량을 측정하였다.In order to investigate the changes in the components of the hydrothermal extract of the mud mushroom fruiting body of the present invention, the anticancer activity, total sugar and reducing sugar content were measured.

진흙버섯 성분에 항암활성이 있음은 이미 공지된 사실로서, 진흙버섯 추출물의 항암활성이 높으면 유효성분의 함량도 높다고 판단할 수 있으므로, 진흙버섯 성분의 대표적 활성으로써 항암활성을 측정하였다.It is known that there is anti-cancer activity in the mud mushroom component, if the anti-cancer activity of the mud mushroom extract can be determined that the content of the active ingredient is high, the anti-cancer activity was measured as a representative activity of the mud mushroom component.

마찬가지로, 진흙버섯의 유효성분의 함량을 반영하는 기준으로 총당 및 환원당의 함량을 측정하여 유효성분의 함량이 가장 높은 최적의 추출조건을 결정하였다.Similarly, by measuring the content of total sugar and reducing sugar as a reference reflecting the content of the active ingredient of the mud mushroom, the optimum extraction condition with the highest content of the active ingredient was determined.

1. 항암활성 측정1. Anticancer activity measurement

항암활성은 쥐의 육종 180 암세포주(murine sarcoma 180 cells)에서 SRB (sulforhodamine B) 방법으로 측정하였다.Anticancer activity was measured by the sulforhodamine B (SRB) method in murine sarcoma 180 cells.

96 웰 플레이트의 각 웰에 1×104의 세포가 포함된 180㎕의 RPMI 1640 배지(페니실린 10,000 units/㎖, 스트렙토마이신 10㎎/㎖ 및 10% FBS)를 가하여 37℃, 5% CO2 가스 존재하에서 24시간 배양하였다. 상기 실시예에서 제조한 진흙버섯 열수 추출물을 PBS로 희석하여 5가지 농도로 만들고 각각 20 ㎕씩 배지에 가하여 48시간 더 배양하였다. 배양이 종료된 후 50 ㎕의 차가운 50% TCA(trichloroacetic acid)를 위에서부터 천천히 가해 주고 잠시 기다린 후 조심스럽게 냉장고로 옮겨 1시간 동안 충분히 고정시키고, 고정이 끝난 후에는 증류수로 5회 이상 세척하여 0.4% SRB 용액 100㎕를 가하여 상온에서 30분 이상 염색하였다. 염색이 끝난 후 1% 아세트산으로 5회 세척하여 잘 건조시키고, 150㎕의 10mM 완충안된 Tris 용액으로 SRB 염료를 잘 녹여내어 96 웰 플레이트용 마이크로플레이트 리더(microplate reader)로 490㎚의 범위에서 흡광도를 측정하였다.To each well of a 96 well plate, 180 μl of RPMI 1640 medium (10,000 units / ml penicillin, 10 mg / ml streptomycin and 10% FBS) containing 1 × 10 4 cells was added to 37 ° C., 5% CO 2 gas. Incubated for 24 hours in the presence. The mud mushroom hydrothermal extract prepared in the above example was diluted with PBS to 5 different concentrations, and 20 µl of each was incubated for 48 hours. After the incubation, 50 μl of cold 50% TCA (trichloroacetic acid) is slowly added from above, wait for a while, and then carefully transferred to the freezer for 1 hour. After fixing, the solution is washed with distilled water at least 5 times for 0.4 100 µl of% SRB solution was added and stained for at least 30 minutes at room temperature. After staining, wash 5 times with 1% acetic acid to dry well, and dissolve the SRB dye with 150 μl of 10 mM unbuffered Tris solution and absorb the absorbance in the range of 490 nm with a microplate reader for 96 well plates. Measured.

2. 총당 측정2. Total sugar measurement

총당은 글루코스를 표준물질로 하여 안트론 방법(Anthrone method)으로 측정하였다.Total sugar was measured by the Antrone method using glucose as a standard.

상기 실시예에서 제조한 진흙버섯 열수 추출물 200㎕를 안트론 시약(0.1g 안트로/76% 황산) 1㎖와 혼합하였다. 혼합물을 12분동안 가열하고, 분광광도계로 620㎚에서 흡광도를 측정하였다.200 µl of the mud mushroom hydrothermal extract prepared in the above example was mixed with 1 ml of anthrone reagent (0.1 g antro / 76% sulfuric acid). The mixture was heated for 12 minutes and the absorbance was measured at 620 nm with a spectrophotometer.

3. 환원당 측정3. Reducing Sugar Measurement

환원당은 글루코스를 표준물질으로 하여 DNS(dinitrosalicylic acid) 분석으로 측정하였다.Reducing sugar was measured by DNS (dinitrosalicylic acid) analysis using glucose as a standard.

상기 실시예에서 제조한 진흙버섯 열수 추출물 500㎕를 DNS 시약 0.5㎖와 혼합하였다. 혼합물을 5분동안 가열하고, 분광광도계로 575㎚에서 흡광도를 측정하였다.500 µl of the mud mushroom hydrothermal extract prepared in Example was mixed with 0.5 ml of DNS reagent. The mixture was heated for 5 minutes and the absorbance was measured at 575 nm with a spectrophotometer.

항암활성, 총당 및 환원당의 측정 결과는 표 2에 나타내었다.The results of anticancer activity, total sugars and reducing sugars are shown in Table 2.

본 발명의 진흙버섯 자실체의 열수 추출 조건에 따른 항암활성, 총당 및 환원당의 함량Anticancer Activity, Contents of Total Sugar and Reducing Sugar According to Hot Water Extraction Conditions of Mud Mushroom Fruit Body of the Present Invention 항암활성(Y1) (%)Anticancer activity (Y 1 ) (%) 총당(Y2) (㎍/㎖)Total sugar (Y 2 ) (µg / ml) 환원당(Y3) (ppm)Reducing Sugar (Y 3 ) (ppm) 1One 60.560.5 0.670.67 352.2352.2 22 55.255.2 0.760.76 394.5394.5 33 71.771.7 1.291.29 494.5494.5 44 77.777.7 3.073.07 422.2422.2 55 80.080.0 0.460.46 429.7429.7 66 77.977.9 0.500.50 496.3496.3 77 86.786.7 1.591.59 983.0983.0 88 91.191.1 1.711.71 12201220 99 67.367.3 0.900.90 486.4486.4 1010 72.072.0 1.141.14 539.8539.8 1111 66.966.9 0.760.76 384.2384.2 1212 89.289.2 1.841.84 693.1693.1 1313 62.062.0 1.001.00 455.2455.2 1414 78.378.3 0.990.99 746.3746.3 1515 66.366.3 1.161.16 508.7508.7 1616 66.366.3 1.161.16 508.7508.7 총 모델 1차 2차 백터곱 (cross-product)Total model 1st order quadratic cross-product ** *** * NS** *** * NS ** ** NS NS** ** NS NS *** *** NS ***** *** NS ** F-비율F-ratio 15.1315.13 8.548.54 39.0539.05 추출시간 추출온도 추출부피Extraction time Extraction temperature Extraction volume NS ** **NS ** ** NS ** NSNS ** NS NS *** ***NS *** ***

※ *** : p<0.001, ** : p<0.01, * : p<0.05 NS : 유의성 없음.※ ***: p <0.001, **: p <0.01, *: p <0.05 NS: No significance.

표 2에 나타난 바와 같이, 항암활성은 추출온도와 추출부피에서 유의성이 나타났으며(p<0.01), 총당은 추출온도에서 유의성이 나타났다(p<0.01). 또한, 환원당은 추출온도와 추출부피에서 유의성이 나타났다(p<0.001).As shown in Table 2, anticancer activity was significant at extraction temperature and extraction volume (p <0.01), and total sugar was significant at extraction temperature (p <0.01). In addition, reducing sugars were significant at extraction temperature and extraction volume (p <0.001).

따라서, 추출온도와 추출부피가 반응요인에 유의한 영향을 끼치고 있음을 알 수 있다.Therefore, it can be seen that the extraction temperature and the extraction volume have a significant effect on the reaction factors.

실험예 2Experimental Example 2 : 진흙버섯 자실체의 열수 추출물의 항암활성, 총당 및 환원당의 회귀분석 : Anticancer Activity, Regression Analysis of Total Sugar and Reducing Sugar of Hot Water Extract of Fruits

상기 실험예 1에서 측정된 항암활성, 총당 및 환원당의 평균값을 이용하여 표면반응곡선에 대한 회귀분석을 수행하였다.Regression analysis was performed on the surface response curve using the average values of anticancer activity, total sugars and reducing sugars measured in Experimental Example 1.

회귀분석 결과는 표 3에 나타내었고, 최적의 추출반응조건은 표 4에 나타내었다.The regression analysis results are shown in Table 3, and the optimum extraction reaction conditions are shown in Table 4.

항암활성, 총당 및 환원당에 의하여 계산된 회귀계수 값Regression coefficient value calculated by anticancer activity, total sugar and reducing sugar 2차 반응식2nd reaction 회귀계수(R2)Regression coefficient (R 2 ) 유의성valence 항암활성(Y1, %)Anticancer activity (Y 1 , %) Y 1 = 639.535** - 5.5775X 1 - 13.5985*X 2 + 2.4615X 3 - 0.2125X 1 2 + 0.11125X 1 X 2 + 0.0755*X 2 2 + 0.01X 1 X 3 - 0.01725X 2 X 3 - 0.0035X 3 2 Y 1 = 639.535 **-5.5775 X 1- 13.5985 * X 2 + 2.4615 X 3 - 0.2125 X 1 2 + 0.11125 X 1 X 2 + 0.0755 * X 2 2 + 0.01 X 1 X 3 - 0.01725 X 2 X 3 - 0.0035 X 3 2 0.95780.9578 0.00180.0018 총당(Y2, ㎍/㎖)Total sugar (Y 2 , Μg / ml) Y 2 = 20.10856-0.508892X 1 - 0.478961X 2 + 0.180609X 3 - 0.007974X 1 2 + 0.011063X 1 X 2 + 0.002481X 2 2 - 0.010688X 1 X 3 - 0.000738X 2 X 3 - 0.000569X 3 2 Y 2 = 20.10856-0.508892 X 1- 0.478961 X 2 + 0.180609 X 3 - 0.007974 X 1 2 + 0.011063 X 1 X 2 + 0.002481 X 2 2 - 0.010688 X 1 X 3 - 0.000738 X 2 X 3 - 0.000569 X 3 2 0.92760.9276 0.00840.0084 환원당(Y3, ppm)Reducing Sugar (Y 3 , ppm) Y 3 = 2908.373276 - 4.151293X 1 - 40.662466X 2 - 158.673603***X 3 - 2.62931X 1 2 + 0.34875X 1 X 2 + 0.150328X 2 2 + 2.085X 1 X 3 + 1.383750***X 2 X 3 + 0.771328*X 3 2 Y 3 = 2908.373276 - 4.151293 X 1 - 40.662466 X 2 - 158.673603 *** X 3 - 2.62931 X 1 2 + 0.34875 X 1 X 2 + 0.150328 X 2 2 + 2.085 X 1 X 3 + 1.383 750 *** X 2 X 3 + 0.771328 * X 3 2 0.98320.9832 0.00010.0001

※ *** : p<0.001, ** : p<0.01, * : p<0.05※ ***: p <0.001, **: p <0.01, *: p <0.05

표 3에 나타난 바와 같이, 항암활성의 회귀계수는 0.9578로, 추출시간이 적을수록(p<0.01), 추출온도가 낮을수록(p<0.05) 항암활성은 저하되었다.As shown in Table 3, the regression coefficient of anticancer activity was 0.9578. The anticancer activity decreased as the extraction time was short (p <0.01) and the extraction temperature was low (p <0.05).

그러나 총당의 회귀계수는 0.9276으로, 추출조건에 대하여 유의성이 인정되지 않았다.However, the total sugar regression coefficient was 0.9276, which was not significant for the extraction condition.

또한, 환원당의 회귀계수는 0.9832로, 추출부피가 클수록(p<0.01) 환원당의 농도가 저하되었다.In addition, the regression coefficient of the reducing sugar was 0.9832, and the concentration of the reducing sugar decreased as the extraction volume was larger (p <0.01).

실험예 3Experimental Example 3 : 진흙버섯 자실체의 열수 추출물의 최적의 추출조건 : Optimal Extraction Conditions of Hot Water Extract of Fruits from Mud Mushrooms

반응표면분석법은 그래프로서 최적화를 이루는 한 방법으로, 등고선도를 이용하여 최적조건을 결정할 수 있다. 그러나 각 반응변수들의 반응표면은 정확하게 일치하지 않으므로 적절한 제한조건을 필요로 한다. 따라서, 주어진 실험조건내에서 예비실험을 통해 얻어진 결과를 바탕으로 제한영역을 설정하고, 각 조건이 일치하는 지역을 중첩되는 등고선도(contour map)로 구하였다.Response surface analysis is a method of optimizing graphically, using contour plots to determine optimal conditions. However, the response surface of each response variable does not exactly match, so appropriate constraints are required. Therefore, based on the results obtained through the preliminary experiments within the given experimental conditions, the limited area was set, and the regions matching each condition were obtained by overlapping contour maps.

항암활성을 종속변수(Y1)로 하여 반응표면분석 및 등고선도를 작성하여 도 1에 나타내었다.Response surface analysis and contour plots were made with anticancer activity as the dependent variable (Y 1 ) and are shown in FIG. 1.

총당을 종속변수(Y2)로 하여 반응표면분석 및 등고선도를 작성하여 도 2에 나타내었다.Response surface analysis and contour plots were made with total sugar as the dependent variable (Y 2 ) and are shown in FIG. 2.

환원당을 종속변수(Y3)로 하여 반응표면분석 및 등고선도를 작성하여 도 3에 나타내었다.Response surface analysis and contour plots were made with reducing sugar as the dependent variable (Y 3 ) and are shown in FIG. 3.

본 발명의 진흙버섯 자실체의 열수 추출물의 최적의 추출조건은 표 4에 나타내었다.The optimum extraction conditions of the hydrothermal extract of the mud mushroom fruiting body of the present invention is shown in Table 4.

본 발명의 진흙버섯 자실체의 열수 추출물의 최적의 추출조건Optimal Extraction Conditions of Hot Water Extract of Mud Mushroom Fruit Body of the Present Invention 추출시간(X1) (hr)Extraction time (X 1 ) (hr) 추출온도(X2) (℃)Extraction temperature (X 2 ) (℃) 추출부피(X3) (㎖/g)Extraction volume (X 3 ) (ml / g) 최대반응Response 반응형태Response type 항암활성 (Y1, %)Anticancer activity (Y 1 , %) 10.2410.24 99.4699.46 23.0023.00 85.585.5 안장점 (saddle point)Saddle point 총당 (Y2, ㎍/㎖)Total sugar (Y 2 , Μg / ml) 10.7010.70 99.1399.13 17.9017.90 2.082.08 안장점Saddle 환원당 (Y3, ppm)Reducing Sugar (Y 3 , ppm) 10.2710.27 96.1196.11 27.8027.80 890.6890.6 안장점Saddle

도 1에 나타난 바와 같이, 본 발명의 진흙버섯 열수 추출물의 항암활성은 추출온도와 추출부피가 증가할수록 증가하는 경향을 보여, 추출온도와 추출부피에 비례함을 알 수 있다.As shown in Figure 1, the anticancer activity of the mud mushroom hydrothermal extract of the present invention shows a tendency to increase as the extraction temperature and extraction volume increases, it can be seen that it is proportional to the extraction temperature and extraction volume.

도 2에 나타난 바와 같이, 본 발명의 진흙버섯 자실체의 열수 추출물의 총당은 추출온도가 증가할수록 증가하나 추출용매는 영향을 주지 않았다. 따라서, 총당은 추출온도에만 비례함을 알 수 있다.As shown in Figure 2, the total sugar of the hydrothermal extract of the mud mushroom fruiting body of the present invention increased as the extraction temperature increased, but the extraction solvent did not affect. Therefore, it can be seen that the total sugar is only proportional to the extraction temperature.

도 3에 나타난 바와 같이, 본 발명의 진흙버섯 자실체의 열수 추출물의 환원당은 추출온도와 추출부피가 증가할수록 증가하는 경향을 보여, 추출온도와 추출부피에 비례함을 알 수 있다.As shown in Figure 3, the reducing sugar of the hot water extract of the mud mushroom fruiting body of the present invention shows a tendency to increase as the extraction temperature and the extraction volume increases, it can be seen that the proportion is proportional to the extraction temperature and the extraction volume.

또한, 표 4에 나타난 바와 같이, 항암활성에 대한 진흙버섯 자실체의 열수 추출물의 최적 조건은 추출시간 10.24시간, 추출온도 99.46℃, 추출부피 23.00㎖/g 이며, 85.5%에서 최대반응이 나타났다.In addition, as shown in Table 4, the optimum conditions of the hydrothermal extract of the mud mushroom fruiting body for anticancer activity was 10.24 hours extraction time, 99.46 ℃ extraction temperature, 23.00ml / g extraction volume, the maximum response was 85.5%.

총당에 대한 진흙버섯 자실체의 열수 추출물의 최적의 조건은 추출시간 10.70시간, 추출온도 99.13℃, 추출부피 17.90㎖/g 이며, 2.08㎎/㎖의 농도에서 최대반응이 나타났다.The optimum conditions of hydrothermal extract of mud mushroom fruiting body on total sugar were extraction time of 10.70 hours, extraction temperature of 99.13 ° C, extraction volume of 17.90ml / g, and maximum reaction at the concentration of 2.08mg / ml.

환원당에 대한 진흙버섯 자실체의 열수 추출의 최적 조건은 추출시간 10.27시간, 추출온도 96.11℃, 추출부피 27.80㎖/g 이며, 890.6ppm에서 최대반응이 나타났다.The optimum conditions for hot water extraction of mud mushroom fruiting bodies for reducing sugars were extraction time of 10.27 hours, extraction temperature of 96.11 ℃, extraction volume of 27.80ml / g, and maximum reaction at 890.6ppm.

따라서, 본 발명의 진흙버섯 자실체의 열수 추출물의 최적의 추출조건은 항암활성, 총당 및 환원당을 기준으로 할 경우 추출시간 10~11시간, 추출온도 96~100℃, 추출부피 17~28㎖/g 로 결정할 수 있다.Therefore, the optimal extraction conditions of the hydrothermal extract of the mud mushroom fruiting body of the present invention is based on the anticancer activity, total sugar and reducing sugar extraction time 10 ~ 11 hours, extraction temperature 96 ~ 100 ℃, extraction volume 17 ~ 28mL / g Can be determined.

실험예 4Experimental Example 4 : 본 발명의 진흙버섯 자실체의 열수 추출물이 급성 호흡기 염증 반응 억제에 미치는 효과 : Effect of Hot Water Extract of Mud Mushroom Fruit Body of the Present Invention on Acute Respiratory Inflammatory Response

본 발명의 진흙버섯 자실체의 열수 추출물이, 랫트에서 급성 호흡기 염증 반응 억제에 미치는 효과를 알아보기 위하여 다음과 같은 실험을 수행하였다.In order to determine the effect of the hot water extract of the mud mushroom fruiting body of the present invention on the inhibition of acute respiratory inflammatory response in rats, the following experiment was performed.

1. 백혈구수 측정1. Leukocyte count

24 마리의 스프라-돌리(Sprague-Dawley) 랫트를 3개의 그룹으로 나누어, 대조군은 생리적 식염수를, 실험군 1은 50㎍의 LPS(lipopolysaccharide)를, 실험군 2는 상기 실시예에서 제조한 진흙버섯 자실체의 열수 추출물을 투여한 후 50㎍의 LPS를 랫트의 기관지에 투여하여, 기관지 삼출액에서 백혈구 수를 측정하였다.Twenty-four Sprague-Dawley rats were divided into three groups, the control group of physiological saline, the experimental group 1 of 50 μg of lipopolysaccharide (LPS), and the experimental group 2 of the mud mushroom fruiting bodies prepared in the above examples. After administration of the hydrothermal extract of, 50 μg of LPS was administered to the bronchus of the rat, and the leukocyte count was measured in the bronchial effusion.

백혈구의 수는 급성 호흡기 염증 반응시 지표가 된다.The number of white blood cells is an indicator for acute respiratory inflammatory reactions.

결과는 표 5에 나타내었다.The results are shown in Table 5.

본 발명의 진흙버섯 자실체의 열수 추출물의 총 백혈구수 및 중호성 백혈구수Total leukocyte count and Neutrophil leukocyte count of hydrothermal extract of mud mushroom fruiting body of the present invention 총 백혈구 수(x 106)Total white blood cell count (x 10 6 ) 중호성 백혈구 수(x 106)Neutrophil leukocyte count (x 10 6 ) 대조군Control 0.30±0.10*0.30 ± 0.10 * 0.09±0.06*0.09 ± 0.06 * 실험군 1Experimental group 1 2.44±0.392.44 ± 0.39 1.82±0.341.82 ± 0.34 실험군 2Experiment group 2 0.28±0.14*0.28 ± 0.14 * 0.05±0.03*0.05 ± 0.03 *

※ 수치 : 평균±표준편차, * : p<0.05※ Number: mean ± standard deviation, *: p <0.05

표 5에 나타난 바와 같이, LPS를 투여한 실험군 1에서 총백혈구의 수는 생리적 식염수를 투여한 대조군에 비하여 8배 증가하였다. 그러나 진흙버섯 자실체의 열수 추출물로 전처리한 후, LPS를 투여한 실험군 2에서는 대조군보다도 백혈구의 수가 오히려 감소하였다.As shown in Table 5, the number of total leukocytes in the experimental group 1 administered with LPS increased 8-fold compared to the control group administered with physiological saline. However, after pretreatment with hydrothermal extract of the mud mushroom fruiting body, the number of leukocytes in the experimental group 2 administered with LPS decreased rather than the control group.

또한, 중호성 백혈구 수에 있어서도 총 백혈구 수와 같은 양상의 결과를 보여주었다.In addition, the result of the same pattern as the total leukocyte count was shown in the number of neutrophils.

일반적으로 리포폴리사카라이드는 사람 또는 동물에서 염증반응을 일으켜 여러 질병의 원인이 되며, 호흡기에 염증이 생기면 백혈구 수가 증가하게 된다.In general, lipopolysaccharides cause inflammatory reactions in humans or animals and cause various diseases. When the respiratory tract becomes inflamed, the white blood cell count increases.

따라서, 본 발명의 진흙버섯 자실체의 열수 추출물은 백혈구의 수를 감소시킴으로, 급성 호흡기 염증 억제 효과가 우수함을 알 수 있다.Therefore, the hydrothermal extract of the mud mushroom fruiting body of the present invention reduces the number of leukocytes, it can be seen that the effect of suppressing acute respiratory inflammation.

2. 사이토킨 측정2. Cytokine Measurement

24 마리의 스프라-돌리(Sprague-Dawley) 랫트를 3개의 그룹으로 나누어, 대조군은 생리적 식염수를, 실험군 1은 50㎍의 LPS(lipopolysaccharide)를, 실험군 2는 상기 실시예에서 제조한 진흙버섯 자실체의 열수 추출물을 투여한 후 50㎍의 LPS를 랫트의 기관지에 투여하여, 기관지 삼출액에서 염증매개인자로서 사이토킨 (cytokine)인 조직괴사인자(Tissue Necrotic Factor-α; TNF-α)와 인터루킨-1β (interleukin-1β; IL-1β)의 억제 정도를 측정하였다.Twenty-four Sprague-Dawley rats were divided into three groups, the control group having physiological saline, the experimental group 1 having 50 µg of lipopolysaccharide (LPS), and the experimental group 2 with the mud mushroom fruiting body prepared in the above examples. 50 μg of LPS was administered to the bronchus of rats after administration of the hydrothermal extract of Tissue Necrotic Factor-α (TNF-α) and interleukin-1β (cytokine) as mediators of inflammation in bronchial effusion. interleukin-1β; IL-1β) was measured.

회수된 기관지액은 10분동안 4℃에서 350 xg로 원심분리한 후, 상층액을 측정전까지 -20℃에 보관하였으며, 사이토킨(TNF-α와 IL-1β)의 변화를 효소결합면역흡수분석법 키트(Enzyme-linked immunosorbent assay kits)로 측정하였다. 이때 측정 한계치는 TNF-α는 12 pg/㎖, IL-1β는 32 pg/㎖ 이었다.The collected bronchial fluid was centrifuged at 350 xg at 4 ° C for 10 minutes, and the supernatant was stored at -20 ° C until measurement. The changes in cytokines (TNF-α and IL-1β) were measured by enzyme-linked immunosorbent assay kits. It was measured by (Enzyme-linked immunosorbent assay kits). At this time, the measurement limit was 12 pg / ml for TNF-α and 32 pg / ml for IL-1β.

IL-1β는 급성 호흡기 염증 반응시 지표가 된다.IL-1β is an indicator in acute respiratory inflammatory responses.

측정 결과는 표 6에 나타내었다.The measurement results are shown in Table 6.

본 발명의 진흙버섯 자실체의 열수 추출물의 TNF-α 및 IL-1β의 변화Changes in TNF-α and IL-1β of Hot Water Extracts of the Mud Mushroom Fruiting Body of the Present Invention TNF-α(pg/㎖)TNF-α (pg / ml) IL-1β(pg/㎖)IL-1β (pg / ml) 대조군Control 200±180200 ± 180 220±98**220 ± 98 ** 실험군 1Experimental group 1 1,180±140*1,180 ± 140 * 650±76*650 ± 76 * 실험군 2Experiment group 2 1,360±120*1,360 ± 120 * 380±46**380 ± 46 **

※ 수치 : 평균±표준편차, *,** : p<0.05※ Number: mean ± standard deviation, *, **: p <0.05

표 6에 나타난 바와 같이, TNF-α의 농도는 실험군 1 및 실험군 2에서 대조군보다 증가하여, 본 발명의 진흙버섯 자실체의 열수 추출물이 염증반응에 영향을 미치지 않음을 알 수 있다.As shown in Table 6, the concentration of TNF-α was increased in the experimental group 1 and the experimental group 2 than the control group, it can be seen that the hydrothermal extract of the mud mushroom fruiting body of the present invention does not affect the inflammatory response.

또한, IL-1β의 농도는 실험군 1에서 대조군보다 3배 정도 증가하였고, 실험군 2에서는 대조군보다 1.7배 정도 증가하여, 본 발명의 진흙버섯 자실체의 열수 추출물이 염증반응을 효과적으로 억제함을 알 수 있다.In addition, the concentration of IL-1β was increased three times as compared to the control group in Experimental group 1, 1.7 times as compared to the control group in Experimental group 2, it can be seen that the hydrothermal extract of the mud mushroom fruiting body of the present invention effectively inhibits the inflammatory response. .

하기에 본 발명의 조성물을 위한 제제예를 예시한다.Examples of preparations for the compositions of the present invention are illustrated below.

제제예 1Formulation Example 1 : 약학적 제제의 제조 : Preparation of Pharmaceutical Formulations

1. 산제의 제조1. Preparation of powder

진흙버섯 자실체의 열수 추출물 2g2 g of hot water extract of fruiting bodies of mud

유당 1g1g lactose

상기의 성분을 혼합하고 기밀포에 충진하여 산제를 제조하였다.The above ingredients were mixed and filled in airtight cloth to prepare a powder.

2. 정제의 제조2. Preparation of Tablets

진흙버섯 자실체의 열수 추출물 100㎎100 mg of hot water extract of fruiting bodies of mud

옥수수전분 100㎎Corn Starch 100mg

유 당 100㎎Lactose 100mg

스테아린산 마그네슘 2㎎2 mg magnesium stearate

상기의 성분을 혼합한 후, 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. 캡슐제의 제조3. Preparation of Capsule

진흙버섯 자실체의 열수 추출물 100㎎100 mg of hot water extract of fruiting bodies of mud

옥수수전분 100㎎Corn Starch 100mg

유 당 100㎎Lactose 100mg

스테아린산 마그네슘 2㎎2 mg magnesium stearate

상기의 성분을 혼합한 후, 통상의 캡슐제의 제조방법에 따라서 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

제제예 2Formulation Example 2 : 식품의 제조 : Manufacture of food

본 발명의 진흙버섯 자실체의 열수 추출물을 포함하는 식품들을 다음과 같이 제조하였다.Food containing a hydrothermal extract of the mud mushroom fruiting body of the present invention was prepared as follows.

1. 조리용 양념의 제조1. Preparation of Cooking Seasonings

본 발명의 진흙버섯 자실체의 열수 추출물 20 ~ 95 중량%로 건강 증진용 조리용 양념을 제조하였다.The hot water extract of the mud mushroom fruiting body of the present invention was prepared for cooking spices for health promotion.

2. 토마토 케찹 및 소스의 제조2. Preparation of Tomato Ketchup and Sauce

본 발명의 진흙버섯 자실체의 열수 추출물 0.2 ~ 1.0 중량%를 토마토 케찹 또는 소스에 첨가하여 건강 증진용 토마토 케찹 또는 소스를 제조하였다.Health-promoting tomato ketchup or sauce was prepared by adding 0.2 to 1.0% by weight of hot water extract of the mud mushroom fruiting body of the present invention to tomato ketchup or sauce.

3. 밀가루 식품의 제조3. Manufacturing of Flour Foods

본 발명의 진흙버섯 자실체의 열수 추출물 0.5 ~ 5.0 중량%를 밀가루에 첨가하고, 이 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하여 건강 증진용 식품을 제조하였다.0.5 to 5.0% by weight of hot water extract of the mud mushroom fruiting body of the present invention was added to wheat flour, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare foods for health promotion.

4. 스프 및 육즙(gravies)의 제조4. Preparation of soups and gravy

본 발명의 진흙버섯 자실체의 열수 추출물 0.1 ~ 5.0 중량%를 스프 및 육즙에 첨가하여 건강 증진용 육가공 제품, 면류의 수프 및 육즙을 제조하였다.The hot water extract of the mud mushroom fruiting body of the present invention was added to the soup and broth to prepare meat products for health promotion, soup and noodles of noodles.

5. 그라운드 비프(ground beef)의 제조5. Preparation of Ground Beef

본 발명의 진흙버섯 자실체의 열수 추출물 10 중량%를 그라운드 비프에 첨가하여 건강 증진용 그라운드 비프를 제조하였다.10% by weight of hot water extract of the mud mushroom fruiting body of the present invention was added to ground beef to prepare ground beef for health promotion.

6. 유제품(dairy products)의 제조6. Manufacture of Dairy Products

본 발명의 진흙버섯 자실체의 열수 추출물 5 ~ 10 중량%를 우유에 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.5 to 10% by weight of hot water extract of the mud mushroom fruiting body of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

7. 선식의 제조7. Manufacture of wire

현미, 보리, 찹쌀, 율무를 공지의 방법으로 알파화시켜 건조시킨 것을 배전한 후 분쇄기로 입도 60메쉬의 분말로 제조하였다.Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh using a grinder.

검정콩, 검정깨, 들깨도 공지의 방법으로 쪄서 건조시킨 것을 배전한 후 분쇄기로 입도 60메쉬의 분말로 제조하였다.Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh.

본 발명의 마른진흙버섯 자실체의 추출물을 진공 농축기에서 감압·농축하고, 분무, 열풍건조기로 건조하여 얻은 건조물을 분쇄기로 입도 60메쉬로 분쇄하여 건조분말을 얻었다.The extract of the dried mud mushroom fruiting body of the present invention was reduced and concentrated in a vacuum concentrator, and the dried product obtained by drying with a spray and a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

상기에서 제조한 곡물류, 종실류 및 진흙버섯 자실체의 열수 추출물의 건조분말을 다음의 비율로 배합하여 제조하였다.The dry powders of the hot water extracts of the grains, seeds and mud mushroom fruiting bodies prepared above were prepared by combining the following ratios.

곡물류(현미 30중량%, 율무 15중량%, 보리 20중량%),Cereals (30% by weight brown rice, 15% by weight radish, 20% by weight barley),

종실류(들깨 7중량%, 검정콩 8중량%, 검정깨 7중량%),Seeds (7% by weight perilla, 8% by weight black beans, 7% by weight black sesame),

진흙버섯 자실체의 열수 추출물의 건조분말(3 중량%),Dry powder (3% by weight) of hydrothermal extract of mud mushroom fruiting body,

영지(0.5중량%),Ganoderma lucidum (0.5% by weight),

지황(0.5중량%)Foxglove (0.5 wt%)

제제예 3Formulation Example 3 : 음료의 제조 : Preparation of Beverages

1. 탄산음료의 제조1. Preparation of Carbonated Drinks

설탕 5~10%, 구연산 0.05~0.3%, 카라멜 0.005~0.02%, 비타민 C 0.1~1%의 첨가물을 혼합하고, 여기에 79~94%의 정제수를 섞어서 시럽을 만들고, 상기 시럽을 85~98℃에서 20~180초간 살균하여 냉각수와 1:4의 비율로 혼합한 다음 탄산가스를 0.5~0.82%를 주입하여 본 발명의 진흙버섯 자실체의 열수 추출물을 함유하는 탄산음료를 제조하였다.5-10% of sugar, 0.05-0.3% citric acid, 0.005-0.02% caramel, 0.1-1% of vitamin C are mixed, and 79-94% purified water is mixed to make syrup, and the syrup is 85-98 Sterilizing at 20 ℃ for 180 seconds, mixed with a cooling water 1: 1 ratio, and then injected with 0.5 ~ 0.82% carbon dioxide gas to prepare a carbonated beverage containing a hot water extract of the fruiting body of the mud mushroom of the present invention.

2. 건강음료의 제조2. Manufacture of health drinks

액상과당(0.5%), 올리고당(2%), 설탕(2%), 식염(0.5%), 물(75%)과 같은 부재료와 진흙버섯 자실체의 열수 추출물을 균질하게 배합하여 순간 살균을 한 후 이를 유리병, 패트병 등 소포장 용기에 포장하여 건강음료를 제조하였다.After sterilizing by homogeneously mixing hot water extract of mud mushroom fruit body with subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%) This was packaged in small packaging containers such as glass bottles and plastic bottles to prepare a healthy beverage.

3. 야채쥬스의 제조3. Preparation of Vegetable Juice

본 발명의 진흙버섯 자실체의 열수 추출물 5g을 토마토 또는 당근 쥬스 1,000㎖에 가하여 건강 증진용 야채쥬스를 제조하였다.5 g of the hydrothermal extract of the fruiting body of the mud mushroom of the present invention was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

4. 과일쥬스의 제조4. Preparation of Fruit Juice

본 발명의 진흙버섯 자실체의 열수 추출물 1g을 사과 또는 포도 쥬스 1,000㎖에 가하여 건강 증진용 과일쥬스를 제조하였다.1 g of hot water extract of the fruiting body of the mud mushroom of the present invention was added to 1,000 ml of apple or grape juice to prepare fruit juice for health promotion.

본 발명의 진흙버섯 자실체의 열수 추출물은 백혈구의 수를 감소시키고, 사이토킨의 농도를 현저히 감소시킴으로, 급성 호흡기 염증 억제 효과가 우수함을 알 수 있다.The hydrothermal extract of the mud mushroom fruiting body of the present invention reduces the number of white blood cells and significantly reduces the concentration of cytokines, and thus it can be seen that the acute respiratory inflammatory effect is excellent.

따라서, 본 발명의 조성물은 감기, 급성 인두염, 급성 비염, 급성 부비강염, 급성 편도염, 급성 후두염, 급성 후두개염 및 급성 기관지염, 급성 상기도 감염증 및 급성 호흡 궁박증후군과 같은 급성 호흡기 질환의 예방 및 치료에 유용하게 사용될 수 있다.Therefore, the composition of the present invention is for the prevention and treatment of acute respiratory diseases such as cold, acute pharyngitis, acute rhinitis, acute sinusitis, acute tonsillitis, acute laryngitis, acute laryngitis and acute bronchitis, acute upper respiratory tract infection and acute respiratory distress syndrome. It can be usefully used.

Claims (2)

펠리너스 바우미 자실체의 열수 추출물을 유효성분으로 하는 급성 호흡기 질환에 유용한 식품 조성물.A food composition useful for acute respiratory diseases comprising hot water extract of Felinus Baumi fruiting body. 제 1항에 있어서, 상기 펠리너스 바우미 자실체의 열수 추출물은 최적의 추출조건으로 추출시간 10~11시간, 추출온도 96~100℃, 추출부피 17~28㎖/g 인 반응표면분석법을 이용하여 추출한 것임을 특징으로 하는 급성 호흡기 질환에 유용한 식품 조성물.According to claim 1, wherein the hot water extract of the Felinus Baumi fruiting body is an optimal extraction condition using a reaction surface analysis method of extraction time 10 ~ 11 hours, extraction temperature 96 ~ 100 ℃, extraction volume 17 ~ 28mL / g Food composition useful for acute respiratory disease, characterized in that the extract.
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CN104800597A (en) * 2015-05-23 2015-07-29 周末 Traditional Chinese medicine composition for treating sphagitis
CN105055882A (en) * 2015-08-19 2015-11-18 周国生 Traditional Chinese medicine for treating acute epiglottitis

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CN104258340A (en) * 2014-09-29 2015-01-07 洛阳御平国生物科技有限公司 Formula of traditional Chinese medicine for treating laryngitis
CN104800597A (en) * 2015-05-23 2015-07-29 周末 Traditional Chinese medicine composition for treating sphagitis
CN105055882A (en) * 2015-08-19 2015-11-18 周国生 Traditional Chinese medicine for treating acute epiglottitis

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