KR100392775B1 - Alismatis Rhizoma tea of rosted type and process for prepatration thereof - Google Patents

Alismatis Rhizoma tea of rosted type and process for prepatration thereof Download PDF

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KR100392775B1
KR100392775B1 KR10-2000-0078050A KR20000078050A KR100392775B1 KR 100392775 B1 KR100392775 B1 KR 100392775B1 KR 20000078050 A KR20000078050 A KR 20000078050A KR 100392775 B1 KR100392775 B1 KR 100392775B1
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South Korea
Prior art keywords
gospel
extract
taxa
tea
taxi
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KR10-2000-0078050A
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Korean (ko)
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KR20020048777A (en
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서권일
조영숙
박정로
이성태
박채규
장진규
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순천시
조영숙
박정로
서권일
이성태
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23FCOFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00Tea; Tea substitutes; Preparations thereof
    • A23F3/16Tea extraction; Tea extracts; Treating tea extract; Making instant tea
    • A23F3/18Extraction of water soluble tea constituents
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23FCOFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00Tea; Tea substitutes; Preparations thereof
    • A23F3/16Tea extraction; Tea extracts; Treating tea extract; Making instant tea
    • A23F3/22Drying or concentrating tea extract
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23FCOFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00Tea; Tea substitutes; Preparations thereof
    • A23F3/16Tea extraction; Tea extracts; Treating tea extract; Making instant tea
    • A23F3/30Further treatment of dried tea extract; Preparations produced thereby, e.g. instant tea
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B65CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
    • B65DCONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
    • B65D85/00Containers, packaging elements or packages, specially adapted for particular articles or materials
    • B65D85/70Containers, packaging elements or packages, specially adapted for particular articles or materials for materials not otherwise provided for
    • B65D85/804Disposable containers or packages with contents which are mixed, infused or dissolved in situ, i.e. without having been previously removed from the package
    • B65D85/808Disposable containers or packages with contents which are mixed, infused or dissolved in situ, i.e. without having been previously removed from the package for immersion in the liquid to release part or all of their contents, e.g. tea bags
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00Food ingredients
    • A23V2250/20Natural extracts
    • A23V2250/21Plant extracts
    • A23V2250/214Tea

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  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Mechanical Engineering (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

본 발명은 택사복음차 및 그 제조방법에 관한 것으로 항균활성, 항산화 효과 및 면역활성이 뛰어난 택사를 건조한 뒤 복음기를 사용하여 복음처리한 후 알류미늄호일 티백(Al-foil tea bag)에 포장함으로써 일반 음용형으로 제조하거나 복음처리된 택사를 분쇄하여 부직포 티백(Tea bag)에 포장하여 침출형 차를 제조함으로써 복음처리하지 않은 택사에 비해 기호도가 증가되고 음용이 간편한 택사복음차를 제공하는 뛰어난 효과가 있다.The present invention relates to a taxa Gospel tea and a method for manufacturing the same, the general drink by packaging the aluminum foil tea bags (Al-foil tea bag) after the Gospel treatment using the Gospel after drying the taxis excellent in antimicrobial activity, antioxidant effect and immune activity By manufacturing leached tea by crushing the processed or gospel-processed tacks in non-woven tea bags, there is an excellent effect of increasing the taste and easy-to-drink taxi teas compared to the non-gospel-free tacks. .

Description

택사복음차 및 그 제조방법{Alismatis Rhizoma tea of rosted type and process for prepatration thereof}Gospel tea of Texas and its manufacturing method {Alismatis Rhizoma tea of rosted type and process for prepatration

본 발명은 택사복음차 및 그 제조방법에 관한 것이다. 더욱 상세하게는, 본 발명은 항균활성, 항산화 효과 및 면역활성효과가 뛰어난 택사를 건조시킨 뒤 복음기를 사용하여 복음처리함으로써 기호도가 증가된 택사복음차 및 그 제조방법에 관한 것이다.The present invention relates to a taxa gospel and a manufacturing method thereof. More specifically, the present invention relates to a taxa Gospel tea and a method of manufacturing the same by increasing the preference by drying the tackiness excellent in antimicrobial activity, antioxidant effect and immune activity effect and then using the Gospel.

택사과(澤瀉科;Alismataceae)식물인 택사(Alismatis Rhizoma)는 단자엽 식물중 진화하는 과정에서 분화된 잔존군으로 세계적으로 약 13속 90여종이 분포되어 있는데, 중국의 복건, 강서 사천, 귀천 및 운남지방에서도 많이 생산되고 있으며, 우리나라의 경우에는 그 생산량의 50% 이상을 차지하고 있는 전남 순천 지방을 비롯하여 전남 무안, 구례 및 경북 상주 지역에서 생산되고 있으며, 생약으로 쓰이는 택사는 질경이 택사(Alisma plantago-aquatica L. var. orientaleSamuels)가 주로 재배 생산된다. 택사의 함유성분은 전분 25%, 단백질 7%, 정유의 furfural, resin, 무기염과 triterpenoid계의 alisol A, B, C와 그 아세테이트(acetate) 및 당류로 알려져 있다. 이와 같은 택사는 예로부터 수분을 제거하고 오장을 도우며 기력을 튼튼히 하는 약재로 알려져 있으며, 칼륨(K) 많이 함유되어 있어 택사를 복용하게되면 뇨중의 칼륨(K)의 배출이 현저히 증가된다. 또한 택사는 혈중 콜레스테롤을 감소시키고, 혈압 및 혈중의 당을 낮추는 작용이 있고 사염화 탄소로 유발된 간독성에 유효한 보간작용이 있음이 보고된 바 있다. 또한 최근에는 택사분말이 무좀에도포약으로 쓰이고 있어 택사에는 다양한 생리활성물질이 존재하는 것으로 생각된다. 그러나 택사의 생리활성에 대한 실제적인 연구결과는 매우 부족하며 이를 이용한 건강식품에 대한 개발도 미흡한 실정이다. Alismatis Rhizoma , an Alismataceae plant, is a remnant group differentiated during the evolution of monocotyledonous plants, with about 13 genera and 90 species distributed worldwide. Fujian, Jiangxi, Sichuan, Guitian and It is also produced in Yunnan province, and in Korea, it is produced in the Jeonnam Suncheon region, which accounts for more than 50% of its production, in Muan, Gurye, and Sangju, Gyeongsangbuk-do, and the taxa used as a herbal medicine is Alisma plantago- aquatica L. var. orientale Samuels) is mainly grown and produced. The constituents of taxa are known as 25% starch, 7% protein, furfural, resin, essential salts, alisol A, B, C, and acetates and acetates of triterpenoids. Such taxa is known as a medicine that removes moisture, helps the intestine, and strengthens energy. It is contained a lot of potassium (K), and when the taxi is taken, potassium (K) in the urine is significantly increased. Taxa has also been reported to reduce blood cholesterol, lower blood pressure and blood sugar, and have an effective interpolation effect on hepatotoxicity induced by carbon tetrachloride. In addition, recently, the tactile powder has been used as a medicine for athlete's foot, so it is thought that various physiologically active substances are present in the tactical taxi. However, the actual research results on physiological activity of taxis are very insufficient and the development of health foods using them is insufficient.

본 발명자들은 상기와 같은 점을 착안하여 순천지역에서 생산되는 택사의 항균, 항산화 활성 및 면역반응 조절작용에 미치는 영향 등과 같은 생리활성을 규명하고 건조 택사를 복음처리하여 택사복음차를 제조함으로써 본 발명을 완성하였다.The inventors of the present invention, by identifying the above-mentioned physiological activity, such as the effect on the antibacterial, antioxidant activity and immune response control of the taxi cab produced in Suncheon region, and the process of tackling the dry taxi cab to produce the gospel tea Was completed.

따라서, 본 발명의 목적은 택사복음차의 제조방법을 제공하는 것이다. 본 발명의 다른 목적은 택사복음차를 제공하는 것이다.Accordingly, it is an object of the present invention to provide a method for producing a taxa gospel gospel. Another object of the present invention is to provide a taxa gospel difference.

도 1은 본 발명의 바람직한 제조방법의 실시예를 공정별로 도시한 개략도이다.1 is a schematic diagram showing an embodiment of a preferred manufacturing method of the present invention for each process.

도 2은 본 발명 택사추출물의 항균활성 측정을위해 시험균주에 택사 물추출물을 첨가한후 생성된 생육저해환을 나타내는 평판배지의 사진도이다.Figure 2 is a photograph of a plate medium showing the growth inhibition ring produced after the addition of the tax water extract to the test strain to determine the antimicrobial activity of the tax extract of the present invention.

도 3a는 본 발명 택사추출물의 바실러스 서브틸러스(B. subtilis)균주의 성장 억제효과를 나타낸 그래프이다.Figure 3a is a graph showing the growth inhibitory effect of the Bacillus subtilis strain of the present invention tack extract.

도 3b는 본 발명 택사추출물의 슈도모나스 아에리그노사(Pseudomonas aerignosa) 균주의 성장 억제효과를 나타낸 그래프이다.Figure 3b is a graph showing the growth inhibitory effect of Pseudomonas aerignosa strain of the present taxa extract.

도 3c는 본 발명 택사추출물의 대장균(E.coli) 균주의 성장 억제효과를 나타낸 그래프이다.Figure 3c is a graph showing the growth inhibitory effect of E. coli strains of the present invention tack extract.

도 3d는 본 발명 택사추출물의 살모넬라 티피무리움(Salmonella typhymuriu- Figure 3d is Salmonella typhymuriu of the present invention taxa extract ( Salmonella typhymuriu-

m)균주의 성장 억제 효과를 나타낸 그래프이다. m ) It is a graph showing the growth inhibitory effect of the strain.

도 3e는 본 발명 택사추출물의 비브리오 파라하모리티커스(Vibrio parahaemo liticus) 균주의 성장 억제효과를 나타낸 그래프이다.Figure 3e is a graph showing the growth inhibitory effect of the Vibrio parahaemo liticus strain of the present taxa extract.

도 4은 택사추출물 처리에 따른 살모넬라 티피무리움(Salmonella typhymuriu Figure 4 is Salmonella typhymuriu according to the process of extracting the taxa ( Salmonella typhymuriu

m)의 전자현미경 사진도이다. m ) electron micrograph.

도 5는 50℃에서 7일 동안 리놀레산 자동산화 과정중 택사추출물(100㎕) 첨가에 따른 과산화물가의 변화를 나타낸 그래프이다.Figure 5 is a graph showing the change in peroxide value according to the addition of the tack extract (100 μl) during linoleic acid automatic oxidation for 7 days at 50 ℃.

도 6는 50℃에서 7일 동안 리놀레산의 자동산화 과정중 택사추출물을 농도별 첨가에 따른 과산화물가의 변화를 나타낸 그래프이다.Figure 6 is a graph showing the change in peroxide value according to the concentration of the tax extract during the automatic oxidation of linoleic acid for 7 days at 50 ℃.

도 7은 본 발명 택사추출물을 쥐간에 처리시 TBA가의 변화를 나타낸 그래프이다.Figure 7 is a graph showing the change in TBA value when the present invention is treated with a rat extract.

도 8a는 본 발명 택사추출물을 비장세포에 처리시 비장세포의 증식에 미치는 영향을 나타낸 그래프이다.Figure 8a is a graph showing the effect on the proliferation of splenocytes when the present invention is treated to splenocytes.

도 8b는 본 발명 택사추출물을 B세포에 처리시 B세포의 증식에 미치는 영향을 나타낸 그래프이다.Figure 8b is a graph showing the effect on the proliferation of B cells when the present invention is treated to B cell tack extract.

도 9a는 본 발명 택사추출물이 동종항원에 대해 반응하는 T세포 증식에 미치는 영향을 나타낸 그래프이다.Figure 9a is a graph showing the effect of the present invention tack extract on T cell proliferation in response to homologous antigen.

도 9b는 본 발명 택사추출물이 동종항원에 대해 반응하는 T세포의 저해도를 나타낸 그래프이다.Figure 9b is a graph showing the degree of inhibition of T cells in response to the tax extract of the present invention homologous antigen.

도 10는 본 발명 택사추출물이 동종항원에 대해 반응하는 T세포의 세포독성에 미치는 영향을 나타낸 그래프이다.Figure 10 is a graph showing the effect of the present invention taxa extract on the cytotoxicity of T cells in response to homologous antigen.

도 11은 본 발명에 의한 택사복음차 제품의 사진도이다.Figure 11 is a photographic representation of a tack gospel product according to the present invention.

본 발명의 상기 목적은 택사 추출물을 제조하여 식중독 및 병원성 미생물에 대한 항균활성, 리놀레산과 간지질에 대한 항산화 효과 및 면역활성을 확인한 뒤 건조된 택사를 복음기를 사용하여 160℃, 30분간 복음처리하여 물에 끊여서 음용할 수 있는 일반형 차와 복음처리한 택사를 분쇄하고 티백(Tea bag)에 포장한뒤 온수에 침출하여 음용할 수 있는 침출용차를 제조함으로써 달성하였다.The object of the present invention is to prepare an extract of the taxa and to determine the antimicrobial activity against food poisoning and pathogenic microorganisms, the antioxidant effect and immune activity against linoleic acid and hepatic lipids, and then dried by using the Gospel to process the Gojigi at 160 ℃, 30 minutes It was achieved by crushing a general type tea and gospel-treated tea crushed in water and packing it in a tea bag and then leaching it in hot water to prepare a leachable tea for drinking.

이하, 본 발명의 구성을 설명한다.Hereinafter, the configuration of the present invention will be described.

본 발명은 택사의 항균, 항산화 및 면역활성을 조사하기 위하여 메탄올, 에탄올 및 물을 사용하여 택사 추출물을 제조하는 단계; 상기 택사추출물의 항균 활성을 조사하기 위하여 식중독 및 병원성 미생물에 대한 항균활성을 측정하고, 택사추출물의 처리에 따른 미생물의 형태변화를 조사하는 단계; 택사추출물의 항산화효과를 조사하기 위하여 리놀레산에 대한in vitro항산화 효과 및 간 지질과산화 억제 기능에 대하여 조사하는 단계; 택사 추출물의 면역세포 증식효과, 대식세포주의 일산화질소 생산에 미치는 영향, 동종항원에 대한 증식반응, 동종항원에 의한 T세포 사이토카인 분비량의 억제 및 동종항원에 대한 T세포의 살세포 작용 억제효과를 조사하는 단계; 건조택사를 복음처리 조건을 달리하여 택사복음차를 제조하고 관능평가, 색도변화 및 택사물추출물의 특성을 조사하여 최적 복음조건을 결정하는 단계; 건조된 택사 100을 복음기에 넣고 20rpm속도로 드럼을 회전시키면서 최적 복음처리 조건인 160℃에서 30분간 복음처리하는 단계; 상기 단계의 복음처리한 택사를 10g씩 Al-foil포장하여 일반음용형으로 제조하는 단계; 복음처리한 택사를 분쇄기를 사용하여 20∼45mesh 크기로 분쇄하고 티백(Tea bag)에 포장하여 침출형 차로 제조하는 단계로 구성된다The present invention comprises the steps of preparing a taxi extract using methanol, ethanol and water to investigate the antibacterial, antioxidant and immune activity of the taxi; Measuring antimicrobial activity against food poisoning and pathogenic microorganisms to investigate the antimicrobial activity of the taxa extract, and examining the morphological changes of the microorganisms according to the treatment of the taxa extract; Irradiating with respect to the in vitro antioxidant activity and lipid peroxidation inhibiting liver functions for the linoleic acid in order to examine the antioxidant activity of the extract Alismataceae; Effects of Taxa Extract on Immune Cell Proliferation, Effects of Macrophage Cells on Nitric Oxide Production, Proliferative Response to Homologous Antigens, Inhibition of T Cell Cytokine Secretion by Homologous Antigens, and T Cell Killer Action on Homologous Antigens Investigating; Manufacturing a taxi grammar by varying gospel processing conditions for dry taxi and determining the optimal gospel condition by investigating sensory evaluation, chromaticity change, and characteristics of taxi extract; Putting the dried taxi tack 100 in the Gospel and rotating the drum at a speed of 20 rpm for 30 minutes at 160 ° C., which is an optimal gospel treatment condition; Al-foil packaging by 10g each of the gospel-treated tack in the step of producing a general drinking type; It is composed of the steps of crushing the processed taeksa into 20 ~ 45mesh size using a grinder and packing it into a tea bag to produce a leachable tea.

본 발명 택사(Alismatis Rhizoma)는 1998년 전남 순천에서 수확한 것을 구입하여 사용하였다. 항균 시험용으로 사용한 균주Bacillus subtilis,Pseudomonas aeruginosa,Escherichia coli,Salmonella typhimurium,Escherichia coliO-157 및Vibrio parahaemolyticus는 한국 종균협회에서 분양 받아 사용하였다. 각종 미생물의 배양에 사용된 배지는 영양 배지(nutrient broth, agar)를 사용하였으며,Vibrio parahaemoliticus는 3% NaCl이 첨가된 배지를 사용하였다. Alismatis Rhizoma of the present invention purchased and used harvested in Suncheon, Jeonnam in 1998. The strains Bacillus subtilis , Pseudomonas aeruginosa , Escherichia coli , Salmonella typhimurium , Escherichia coli O-157 and Vibrio parahaemolyticus were used for antimicrobial testing. Nutrient broth (agar) was used for the culture of various microorganisms, and 3% NaCl was added for Vibrio parahaemoliticus .

택사의 면역활성 측정을 조사하기 위해 사용한 생쥐(BALB/c, C57BL/6)는, 대한실험동물센터(충북 음성군)에서 구입한 후 생후 8∼12주된 암컷을 사용하였으며, 실험 전까지 고형사료와 1차 증류수를 공급하면서 사육하였다. RPMI 1640배지, 안티바이오틱 안티마이코틱은 Gibco BRL(Grand Island, NY, USA)제품을 사용하였으며, FCS(fertal calf serum)은 PAA제품을 사용하였다. LPS와 2-ME(2-mercaptoehtanMice (BALB / c, C57BL / 6) used to investigate the immune activity of taxa were purchased from the Korea Experimental Animal Center (negative group Chungbuk), and 8-12 weeks old females were used. It was bred while supplying tea distilled water. RPMI 1640 medium, antibiotic antimicrobial was used Gibco BRL (Grand Island, NY, USA) products, FCS (fertal calf serum) was used PAA products. LPS and 2-ME (2-mercaptoehtan

ol), NaHCO3, N-1-나프틸-에틸렌-디아민(N-1-naphthyl-ethylen-diamine)과 썰파닐아마이드(Sufanilamide)는 Sigma사 제품을 사용하였다. 또한 세포독성 측정(Cytotoxicity assay)에 사용된 시약(Cyto Tox 96Non-radioactive Cytotoxicity Assay)과 세포증식 측정에 사용된 시약(Cell titer96Aqueous One solution cell proliferation Assay)은 Promega사 제품을 사용하였고, 사이토카인(cytokine)측정에 필요한 항체는 Pharmingen사의 제품을 사용하였다. B 세포 분리에 사용된 Sephadex G-10도 Pharmingen사의 제품을 사용하였다. 세포배양을 위한 세포주로는 생쥐 단핵/대식세포 계열의 세포주인 RAW264.7을 한국세포주은행(서울대학교 의과대학 암연구소)에서 구입하였다. 상기 세포배양을 위한 배지는 RPMI 1640배지에 NaHCO32g/L와 페니실린(penicillin) 100unit/ml, 스트렙토마이신 100mg/ml를 첨가하여 사용하였다.ol), NaHCO 3, N-1-naphthyl-ethylen-diamine and sulfanamide were used by Sigma. In addition, the reagents used in the cytotoxicity assay (Cyto Tox 96). Non-radioactive Cytotoxicity Assay and Reagents Used to Measure Cell Proliferation (Cell titer96) Aqueous One Solution Cell Proliferation Assay was used for Promega, and the antibodies required for cytokine measurement were used for Pharmingen. Sephadex G-10 used for B cell separation was also used by Pharmingen. As a cell line for cell culture, RAW264.7, a mouse mononuclear / macrophage cell line, was purchased from Korea Cell Line Bank (Seoul National University College of Medicine). Medium for the cell culture was used by adding NaHCO 3 2g / L, penicillin 100unit / ml, streptomycin 100mg / ml to RPMI 1640 medium.

항균활성 및 항산화능을 측정하기 위하여 사용한 택사(Alismatis Rhizoma) 추출물은 메탄올, 에탄올 및 물과 같은 용매를 사용하여 택사 100g당 1500ml의 비율로 용매를 첨가하고 환류냉각기를 이용하여 메탄올과 에탄올은 80℃, 물은 100℃에서 2시간 추출한후 택사 g 당 0.5ml가 되도록 농축하여 제조하였다. Alismatis Rhizoma extract used to measure antimicrobial activity and antioxidant activity was added at a rate of 1500 ml per 100 g of taxa using solvents such as methanol, ethanol and water, and methanol and ethanol at 80 ° C using a reflux condenser. , Water was prepared by extracting at 100 ℃ for 2 hours and concentrated to 0.5ml per g of taxi.

본 발명의 택사복음차의 복음처리는 실험실용 복음기(Laboratory roaster)를 사용하였으며 분쇄기는 실험실용 분쇄기(Laboratory Mill, Model 4, Arthur H.Thomas, PA, U.S.A)를 사용하였다. 택사의 복음처리조건은 150∼160℃에서 30∼60분간 이며 바람직한 조건은 160℃, 30분간 처리하는 것이다. 택사를 복음처리한 후 분쇄하지 않고 일반음용형으로 제조한 복음차는 원형의 택사 10g을 물 2ℓ에 첨가한 다음 100℃에서 약 1시간 정도 끊인후 수시로 보리차 대용으로 음용이 가능하다. 또한 침출차용으로 분쇄하여 제조한 택사복음차는 1 티백(Tea bag)을 90℃온수 120㎖에 침출하여 음용할 수 있다. 택사복음차의 포장은 5∼15g씩 Al-foil로 포장하거나 1∼2g씩 티백(Tea bag)에 포장하였다. 상기 포장시 바람직하게는 10g을 Al-foil로 포장하거나 1.4g씩 티백(Tea bag)에 포장하였다.The Gospel treatment of the Gospel of Taxa of the present invention used a laboratory roaster, and a grinder was a laboratory mill (Laboratory Mill, Model 4, Arthur H. Thomas, PA, U.S.A). Gospel treatment condition of taxi is 30-60 minutes at 150-160 degreeC, and preferable conditions are 160 degreeC and 30-minute process. Gospel tea produced in general drinking form without crushing after taksa is added to 10 grams of round talc in 2 l of water, and after about 1 hour at 100 ℃ can be used as a substitute for barley tea. In addition, the tack gospel tea prepared by pulverizing for leaching tea can be brewed by leaching one tea bag (Tea bag) in 120ml of hot water. The packaging of the Gospel of Texas was 5-15g each in Al-foil or 1-2g each in tea bags. In the packaging, preferably 10g was packed in Al-foil or 1.4g each in a tea bag.

본 발명 택사복음차의 바람직한 제조방법은 도 1에 도시하였다.Preferred method of manufacturing the present invention taxi gospel is shown in FIG.

이하, 본 발명의 구체적인 방법을 실시예를 들어 상세히 설명하고자 하지만,본 발명의 권리범위는 이들 실시예에만 한정되는 것은 아니다.Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

실험예 1: 택사 추출물의 항균효과 조사Experimental Example 1: Investigation of antimicrobial effect of taxa extract

택사 추출물의 항균효과를 조사하기 위하여 택사 추출물을 사용하여 항균력, 미생물 생육도에 미치는 영향, 미생물의 형태변화에 미치는 영향을 조사하였다.In order to investigate the antimicrobial effect of taxa extracts, the effects of antimicrobial activity, microbial growth, and microbial morphology were investigated.

항균력의 측정은 Farag의 방법을 변형하였다. 즉, 아가(agar) 1.5%가 함유되어 있는 생육배지를 페트리 디쉬(petri dish)의 밑면에 얇게 펴고 그위에 다시 0.6%의 agar가 함유된 생육배지를 부어 2중의 평판 배지를 만든 후 각 균주를 평판배지에 도말하였다. 상기 균주가 도말된 평판배지에 택사추출물을 일정량 가한 직경 0.8cm의 여지 디스크(disc)를 올려놓고 36℃에서 24시간 배양한 뒤, 생성되는생육저해환을 측정하여 항균력을 조사하였다.The measurement of antimicrobial activity modified Farag's method. That is, the growth medium containing 1.5% agar is thinly spread on the underside of a petri dish, and the growth medium containing 0.6% of agar is poured again to make a double flat medium. The plate was plated. The strain was placed on a plate medium coated with a certain amount of the tack extract extract 0.8cm diameter disk (disc) and incubated at 36 ℃ for 24 hours, the resulting growth inhibition was measured to investigate the antibacterial activity.

미생물의 생육도 측정은 액체배지 희석법을 사용하였다. 시험관에 5ml의 생육배지를 넣고 대수기 중기까지 배양된 균체 배양액 1%를 접종한 후 택사 추출물 함량을 배지 5ml당 30∼200㎕가 되도록 첨가하고 각 균주의 최적 온도에서 배양하면서 경시적으로 미생물의 생육정도를 분광광도계(spectrophotometer)를 사용하여 660nm에서 흡광도를 측정하였다.Microbial growth was measured using the liquid medium dilution method. Inoculate 5 ml of growth medium into the test tube and inoculate 1% of the cell culture cultured up to the middle of the logarithmic season, and then add tac extract extract to 30 to 200 µl per 5 ml of medium and incubate at optimum temperature of each strain. The extent of growth was measured by absorbance at 660nm using a spectrophotometer.

택사(Alismatis Rhizoma) 추출물에 의한 미생물의 형태변화 조사는 식품부패 미생물에 택사 추출물을 처리한 후 전자현미경으로 그 형태 변화를 관찰하였다. 부패미생물로Salmonella typhimurium을 배지에 48시간 배양한 다음 배양 균주 일부에 택사 추출물을 배지 ml당 100㎕로 첨가하고 3시간 방치한 뒤 이를 원심분리하여 균체를 분리한 후 0.05M 인산 완충액(phosphate buffer)으로 희석하고 0.45㎕ membrane filter에 균체를 고정하였다. 이를 5% 글루타알데하이드(glutaaldehyde) 용액에 하룻밤 담구어 멸균수로 세척한 뒤 30∼100%의 에탄올에 차례로 담구어 탈수하고, 이소아밀아세테이트(isoamylacetate)에 약 30분간 담구어 건조시킨 후 전자현미경 촬영시료를 조제하여 관찰하였다.The morphological changes of microorganisms by Alixmatis Rhizoma extracts were treated with food extract microorganisms and their morphological changes were observed by electron microscopy. Incubate Salmonella typhimurium as a decay microorganism in a culture medium for 48 hours, add 100 microliters of taxa extract per ml of culture strain to the culture strain, leave for 3 hours, and centrifuge to isolate the cells. 0.05M phosphate buffer The cells were diluted with and fixed to 0.45 μl membrane filter. Soak overnight in 5% glutaaldehyde (glutaaldehyde) solution, wash with sterile water, and then immerse in 30 ~ 100% ethanol one by one in order to dehydrate, immerse in isoamylacetate for about 30 minutes to dry and electron microscope The photographing sample was prepared and observed.

실험결과, 택사 추출물의 항균 활성은 도 2와 표1에 나타낸 바와 같이, 물추출물이 메탄올 및 에탄올 추출물보다 항균활성이 강하게 나타났다. 또한Escherichia coliO-157균주를 제외한 모든 시험균주에 대하여 항균활성이 강하게 나타났으며 각 시험균주에 대한 택사 추출물의 투명환(clear zone)은Vibrio parahaemolyticus, Bacillus subtilis, Salmonella typhimurium, Pseudomonasaeruginosa 및 Escherichia coli에 대하여 각각 18.0, 16.0, 16.0, 13.0, 11.5mm였으며 특히,Vibrio parahaemolyticus균주에 대해서 항균활성이 가장 강하게 나타났다.Experimental results, as shown in Figure 2 and Table 1, the antimicrobial activity of the taxa extract, water extract showed stronger antibacterial activity than methanol and ethanol extract. Also had antimicrobial activity appeared strongly to all tested strains except for Escherichia coli O-157 strain clear zone (clear zone) of Alismataceae extract of each test strain in Vibrio parahaemolyticus, Bacillus subtilis, Salmonella typhimurium, Pseudomonasaeruginosa and Escherichia coli 18.0, 16.0, 16.0, 13.0, and 11.5mm, respectively, and the antimicrobial activity of Vibrio parahaemolyticus was the strongest.

택사 추출물의 항균활성Antimicrobial Activity of Taxa Extracts 균주Strain 메탄올 추출물Methanol extract 에탄올 추출물Ethanol extract 물 추출물Water extract Bacillus subtilisBacillus subtilis 14.014.0 13.513.5 16.016.0 Pseudomonas aerignosaPseudomonas aerignosa 14.014.0 14.014.0 13.013.0 E.coliE.coli 11.011.0 11.011.0 11.511.5 Salmonella typhymuriumSalmonella typhymurium 14.014.0 13.513.5 16.016.0 E.coliO-157 E.coli O-157 8.08.0 8.08.0 8.08.0 Vibrio parahaemolyticuaVibrio parahaemolyticua 16.016.0 16.516.5 18.018.0 〔주〕단위 :mm(Note) Unit: mm

택사(Alismatis Rhizoma) 추출물이 미생물의 증식에 미치는 영향을 조사한 결과, 도 3a∼3e에 나타낸바와 같이 택사추출물을 배지 5ml당 30㎕만 첨가하여도 균의 성장이 억제되었으며 150㎕ 첨가시는 거의 모든 균에서 대조구에 비하여 50% 이상 균의 성장이 억제되었으며, 200㎕ 첨가시는 균의 성장이 거의 억제되었다.As a result of investigating the effect of Alismatis Rhizoma extract on the growth of microorganisms, as shown in Figs. 3a to 3e, the growth of the bacteria was inhibited even by adding 30 μl of the Taxa extract per 5 ml of the medium. The growth of the bacteria was inhibited by more than 50% compared to the control, and the growth of the bacteria was almost inhibited when 200μl addition.

택사 추출물 처리에 따른 미생물의 형태 변화를 조사한 결과, 도 3에 나타낸 바와 같이Salmonerlla typhimurium에 택사 물추출물을 처리한 후 대조구와 비교시 균체의 표층구조가 허물어지는 심한 형태적 변화를 나타내었다. 서 등(서권일,이상원, 양기호 :한국농산물저장유통학회(1999), 6(1))은E.coliO-157에 산수유 물추출물을 처리한 후 전자현미경으로 관찰한 결과 대조구와 비교하여 산수유 물추출물을 처리한 균체에서 세포막 기능이 파괴되어 세포내용물이 균체 외부로 유출되어균체의 생육이 억제되는 현상을 나타내었다고 보고하였고, 서 등은 대장균에 겨자 물추출물을 처리하였을 때 균체 표면이 수축되고 표층구조가 심하게 허물어졌다고 보고하였는데, 본 결과의 이와 같은 현상들도 상기의 연구보고와 비교하여 볼 때 택사 물추출물에 의한 미생물의 세포벽 및 세포막의 기능이 파괴되어 용균이나 균체성분의 노출로 인한 결과라고 생각된다.As a result of investigating the morphological changes of the microorganisms according to the extract of the taxa extract, as shown in FIG. 3, after treating the taxa water extract with Salmonerlla typhimurium , the surface structure of the cells was severely compared with the control group. Seo et al. (Seo Kwon Il, Sang Won Lee, Ki Ho Ho: Korean Agricultural Products Storage and Distribution Association (1999), 6 (1)), treated with E. coli O-157 extracts of cornus water, were observed by electron microscopy. It was reported that the cell membrane function was disrupted in the cells treated with the extract and the contents of the cells were leaked out of the cells, thereby inhibiting the growth of the cells. It was reported that the structure was severely collapsed.These phenomena are also the result of the exposure of lysates or cell components due to the destruction of the cell wall and cell membrane of microorganisms by taxi water extract. I think.

실험예 2: 택사 추출물의 항산화 효과 조사Experimental Example 2: Investigation of Antioxidant Effect of Taxa Extract

택사 추출물(Alismatis Rhizoma)의 리놀레산(linoleic acid)에 대한in vitro항산화 및 간 지질과산화 억제 기능에 대하여 조사하였다. 택사 추출물의 항산화 효과 측정을 위하여 삼각플라스크에 리놀레산(linoleic acid) 1g, 에탄올(ethanol) 10ml 및 소정의 택사 추출물을 첨가한 후 0.2M 인산완충용액(phosphate buffer) 25ml를 가하여 37℃에서 일정기간 저장한 다음 반응용액을 분액깔대기에 옮겨 클로로포름(chloroform) 25ml를 가하여 2∼3회 반복 추출하였다. 상기 클로로포름(chloroform) 추출액에 아세트산(acetic acid) 25ml와 포화 KI용액 1ml를 가하여 암소에서 5분간 방치한 다음 증류수 50ml를 가하여 1/100 N Na2S2O3용액으로 적정하였다. 택사추출물은 각각 메탄올, 에탄올, 물추출물을 첨가하였고, BHT 0.1%를 첨가하여 50℃에서 7일간 저장하면서 과산화물가를 측정하였다.The in vitro antioxidant and hepatic lipid peroxidation inhibitory effects on linoleic acid of Alismatis Rhizoma were investigated. In order to measure the antioxidant effect of taxa extract, 1 g of linoleic acid, 10 ml of ethanol, and a predetermined taxa extract were added to a Erlenmeyer flask, and 25 ml of 0.2 M phosphate buffer was added and stored at 37 ° C. for a period of time. Then, the reaction solution was transferred to a separatory funnel, and 25 ml of chloroform was added thereto, followed by extraction two to three times. 25 ml of acetic acid and 1 ml of saturated KI solution were added to the chloroform extract, which was allowed to stand in the dark for 5 minutes, and then 50 ml of distilled water was added and titrated with a 1/100 N Na 2 S 2 O 3 solution. Taxi extract was added methanol, ethanol, and water extract, respectively, and BHT 0.1% was added and stored at 50 ° C. for 7 days to measure peroxide value.

택사 추출물의 지질과산화 억제효과는 흰쥐의 간 균질액(liver homogenate)를 사용하여in vitro로 조사하였다. 흰쥐의 간을 적출하여 인산완충용액(phospha-Lipid peroxidation inhibitory effect of Taxa extract was investigated in vitro using liver homogenate in rats. Phosphate buffer solution (phospha-

te buffer, pH 7.4)로 균질화한 다음 균질액에 과산화수소(H2O2) 1M과 50mM의 황산철(FeSO4) 및 택사 추출물 0.05ml를 가하여 42℃에서 72시간 동안 저장한 후 생성된 TBARS(thiobarbituric acid reactive substances)함량을 측정하였다.After homogenization with te buffer, pH 7.4), 1M of hydrogen peroxide (H 2 O 2 ), 50 mM iron sulfate (FeSO 4 ), and 0.05 ml of taxa extract were added to the homogenate, and stored at 42 ° C. for 72 hours. thiobarbituric acid reactive substances) was measured.

실험결과, 도 5에 나타낸 바와 같이 택사 메탄올 추출물, 에탄올 추출물 및 물추출물 100㎕ 첨가시 대조구에 비하여 과산화가가 모두 낮게 나타났으며, 물추출물의 과산화물가는 메탄올 및 에탄올 추출물보다 작았으나, 0.1% BHT 첨가시 보다는 조금 높게 나타났다. 또한 택사 물추출물을 50, 100, 150 및 200㎕의 농도로 첨가한 후 과산화물가를 측정한 결과 도 6에 나타낸 바와 같이 모두 대조구에 비하여 매우 낮게 나타났으며, 150㎕ 첨가시는 5일 저장시까지 과산화물이 거의 생성되지 않았는데 이는 0.1% BHT 100㎕를 첨가한 경우와 비슷한 정도의 효과였다. 권 등(권오근, 손진창, 김상철, 정신교, 박승우 :한국농산물저장유통학회(1998), 5(281))은 목단피 메탄올 추출물의 에칠아세테이트 분획의 리놀레산(linoleic acid)에 대한 항산화 효과가 87.3%이었고, 비교구인 BHT는 96.2%로 나타났다고 보고하였고, 서 등(서권일, 이상원, 양기호 :한국농산물저장유통학회(1999), 6(1))은 리놀레산(linoleic acid)에 대하여 산수유 물추출물을 10, 30 및 50㎕ 첨가하고 50℃에서 7일간 저장한 후 과산화물가를 측정한 결과 대조구의 187meq/kg에 비하여 42, 30 및 25meq/kg으로 나타나 상당한 항산화 효과가 있는 것으로 보고하였는데, 본 실험결과 택사 물추출물을 50㎕만 첨가하여도 상당한 항산화 효과가 있는 것으로 나타났다.As a result, as shown in FIG. 5, when the methanol extract, ethanol extract, and water extract 100 μl were added, the peroxide value was lower than that of the control, and the peroxide value of the water extract was smaller than that of the methanol and ethanol extracts, but was 0.1% BHT. A little higher than when added. In addition, as a result of measuring the peroxide value after adding the water extracts of the taxi water at the concentrations of 50, 100, 150 and 200 μl, as shown in FIG. 6, all of them were very low compared to the control, and when 150 μl was added, the peroxide was stored until 5 days storage. This was hardly produced, which was about the same as adding 100 μl of 0.1% BHT. Kwon et al. (Kwon Oh-Geun, Son Jin-Chang, Sang-Chul Kim, Sang-Cheol Kim, Pyo-Sang, Park, Seung-Woo: Korean Society of Agricultural Products and Storage (1998), 5 (281)) showed that the antioxidant activity of linoleic acid in the ethyl acetate fraction of methanol extract was The comparison group reported that BHT was 96.2%, and Seo et al. (Seo, Kwon-Il, Sang-Won Lee, and Ki-Ho Yang: Korean Agricultural Products Storage and Distribution Association (1999), 6 (1)) showed that the extracts of linusic acid were 10, 30 and After adding 50 μl and storing at 50 ° C. for 7 days, the peroxide value was measured to be 42, 30 and 25 meq / kg compared to 187 meq / kg of the control, which showed a significant antioxidant effect. Addition of only μl showed significant antioxidant effect.

흰쥐의 간 지질에 대한 TBA가는 도 7에 나타낸 바와같이 택사 물추출물울 첨가하지 않은 대조구에서 3597μmol malondialdehyde(MDA)/g liver이었고, 택사 물추출물을 첨가한 시험구들의 TBA가는 1443 μmol MDA/g liver로서 모두 대조구에 비하여 낮게 나타나 항산화 효과가 있었다.The TBA value of liver lipids in rats was 3597 μmol malondialdehyde (MDA) / g liver in the control group without the addition of tax water extract as shown in FIG. 7, and the TBA value of the test groups to which the water extract was added was 1443 μmol MDA / g liver. All showed lower anti-oxidant effect than the control.

실험예 3: 택사 추출물의 면역활성 측정Experimental Example 3: Determination of Immune Activity of Taxa Extract

택사(Alismatis Rhizoma) 추출물의 면역세포 증식효과, 대식세포주의 일산화질소 생산에 미치는 영향, 동종항원에 대한 증식반응, 동종항원에 의한 T세포 사이토카인(cytokine) 분비량의 억제, 동종항원에 대한 T세포의 살세포 작용 억제효과 등을 조사하였다. 생쥐의 비장세포 분리를 위하여 생쥐(BALB/c)를 경추탈골로 희생시킨 후, 알콜로 소독하여 해부대에 올려놓고 오른쪽 옆구리쪽을 절개하여 비장을 떼어 내었다. 상기 비장을 핀셋을 이용하여 단일세포(single cell)로 만들고 세척용 배지로 3회 세척후 적당량의 10% FCS-RPMI 1640 배지에 희석하여 사용한다. 상기 분리된 비장세포를 10ml의 10% FCS-RPMI 1640에 희석한 후 40㎕ anti-Thy1.2 ascites를 넣고 4℃에서 60분간 배양하였다. 그 후에 4℃, 1200rpm에서 6분간 원심침전하여 5ml의 FCS-RPMI 1640에 희석하고 250㎕ rabbit complement를 첨가하여 37℃ 수욕에서 45분간 배양하였다. 그 다음 10ml 세척용 배지로 4℃, 1200 rpm에서 6분간 원심침전을 두 번 반복하고 10% FCS-RPMI 1640 배지 500㎕에 희석하여 세파독스 G-10(Sephadox G-10) 컬럼을 통과시켜서 B세포만을 순수분리 하였다. T세포의분리는 상기 분리된 비장세포를 10% FCS-RPMI 1640 1ml에 희석하여 Nylon wool 컬럼에 넣고 37℃, 5% CO2배양기에 60분간 배양하였다. 배양후 나일론 울(Nylon wool) 컬럼을 37℃로 미리 데워진 배지로 세척하여 비부착성인 T세포만을 순수 분리하여 alloreative T세포로 사용하였다.Effects of Alismatis Rhizoma Extract on the Proliferation of Immune Cells, Effects of Macrophage Cells on Nitric Oxide Production, Proliferative Response to Homologous Antigens, Inhibition of T Cell Cytokine Secretion by Homologous Antigens, and T Cells Against Homogenous Antigens The inhibitory effect of killer cells was investigated. To separate the splenocytes of the mice, the mice (BALB / c) were sacrificed with cervical distal bone, sterilized with alcohol, placed on the dissection, and the spleen was removed by dissecting the right flank. The spleen is made into single cells using tweezers, washed three times with washing medium, and diluted in an appropriate amount of 10% FCS-RPMI 1640 medium. The isolated splenocytes were diluted in 10 ml of 10% FCS-RPMI 1640, and 40 µl anti-Thy1.2 ascites were added thereto and incubated at 4 ° C. for 60 minutes. Thereafter, the mixture was centrifuged at 4 ° C and 1200rpm for 6 minutes, diluted in 5ml of FCS-RPMI 1640, and incubated for 45 minutes in a 37 ° C water bath by adding 250µl rabbit complement. Then, centrifugation was repeated twice for 6 minutes at 4 ° C. and 1200 rpm with 10 ml washing medium, diluted in 500 μl of 10% FCS-RPMI 1640 medium, and passed through a Sephadox G-10 column. Only cells were purified. Separation of T cells was diluted in 1 ml of 10% FCS-RPMI 1640 and the splenocytes were separated into nylon wool column and incubated for 60 minutes in 37 ℃, 5% CO 2 incubator. After incubation, the nylon wool column was washed with media preheated to 37 ° C., and only non-adherent T cells were purified and used as alloreative T cells.

비장세포, 순수 분리한 B세포, 또는 순수 분리한 T세포에 allogenic 비장세포(C57BL/6)을 96 well plate에 넣고 여기에 택사 추출물을 1, 10, 100, 1000㎍/ml농도로 넣어 배양한 다음 2일, 3일 4일째에 각 세포의 증식정도를 측정하였다. 측정은 Cell titer96Aqueous One Solution Cell proliferation Assay(promega)를 사용하여 2일, 3일, 4일째 각각 배양된 배양액 100㎕에 세포역가측정용 시약(Cell titer) 15㎕씩 첨가하고 4∼8시간 동안 배양한 다음 490nm에서 O.D값을 측정하였다.Splenocytes, purely isolated B cells, or purely isolated T cells were cultured in 96 well plates with allogenic splenocytes (C57BL / 6) in 1, 10, 100, 1000 ㎍ / ml concentrations. Next, the proliferation of each cell was measured on day 2 and day 3 and day 4. Measure the cell titer96 15 μl of cell titer was added to 100 μl of the culture solution on day 2, 3, and 4 using Aqueous One Solution Cell proliferation Assay (promega), followed by incubation for 4 to 8 hours, followed by 490 nm. OD value was measured at.

택사 추출물이 대식세포주의 일산화질소 생산에 미치는 효과를 알아보기 위하여 택사 추출물을 1, 10, 100, 1000㎍/㎖의 농도로 단독 처리하거나 또는 LPS, IFN-γ를 같이 첨가하였을 때, 대식세포가 생산하는 일산화질소의 농도를 측정하였다. 안정된 일산화질소(NO) 산화물인 NO2 -(nitrite)는 Griess 반응을 이용하여 측정하였다. 대식세포주(RAW264.7)를 적당한 조건하에서 48시간 배양한 후 배양 상층액을 96well plate에 100㎕씩 넣고 여기에 Greiss시약 (0.1%N-1-naphthyl-ethylendiamine in H2O + 1% sulfanilamide in 5% H3PO4)을 동량 첨가하여 10분간 반응시킨 후, 마이크로플래이트 리더(microplate reader, TitertekMultiscan Plus, Finland)로 570nm에서 흡광도를 측정하였다. 질산염(Nitrate)의 농도는 질산 나트륨(sodium nitrite)을 32μM에서부터 0.25μM까지 2배씩 희석하여 얻은 표준 곡선과 비교하여 계산하였다.To investigate the effects of taxa extract on nitric oxide production in macrophages, macrophages were treated with 1, 10, 100, and 1000 µg / ml concentrations, or when LPS and IFN-γ were added. The concentration of nitric oxide produced was measured. Stable nitrogen monoxide (NO) oxide, NO 2 - (nitrite) was measured using the Griess reaction. Incubate the macrophage line (RAW264.7) under appropriate conditions for 48 hours, add 100 µl of the culture supernatant to a 96well plate, and add Greiss reagent (0.1% N -naphthyl-ethylendiamine in H 2 O + 1% sulfanilamide in After reacting for 10 minutes by adding the same amount of 5% H 3 PO 4 ), the absorbance was measured at 570 nm with a microplate reader (Titertek Multiscan Plus, Finland). The concentration of nitrate was calculated by comparison with a standard curve obtained by diluting sodium nitrite from 32 μM to 0.25 μM in 2-fold.

택사 추출물이 동종항원에 의한 증식반응에 어떠한 효과를 나타내는지 알아보기 위하여, 동종항원을 유도하기 위한 세포수 결정실험을 수행하였다. 표적세포(Target cells)인 MMC를 처리한 생쥐 C57BL/6의 비장세포와 효과세포(Effector cells)로 비장세포에서 분리한 T세포(생쥐 BABL/c의 T세포)를 함께 3일동안 배양하여 최대 증식반응을 나타내는 조건을 확인하였다.In order to investigate the effects of taxa extract on the proliferative response by homologous antigens, cell number determination experiments were performed to induce homologous antigens. Splenocytes of mouse C57BL / 6 treated with MMC, which are target cells, and T cells (T cell of mouse BABL / c), which were isolated from splenocytes with effector cells, were cultured together for 3 days. The conditions showing the proliferative response were confirmed.

동종항원에 의한 T세포의 증식반응이 택사 추출물에 의하여 유의하게 억제된 원인을 알아보기 위하여, 동종항원에 반응하는 T세포가 분비하는 사이토카인(cytokine)의 생산량을 측정하였다. T세포가 반응할 때 분비하는 사이토카인(cytokine)의 양은 비장세포와 택사를 같이 배양한 상층액을 24시간 후에 회수하여 상층액에 포함된 IL-2, IL-4, IFN-γ,IL-10의 양을 ELISA(enzyme linked immounso-In order to determine the cause of the proliferation of T cells caused by allogeneic antigens significantly inhibited by the taxa extract, the amount of cytokines produced by T cells in response to allogeneic antigens was measured. The amount of cytokines secreted when T cells reacted was recovered after 24 hours of supernatant cultured with splenocytes and taxa, and IL-2, IL-4, IFN-γ, IL- contained in the supernatant. The amount of 10 ELISA (enzyme linked immounso-

rbent assay)kit로 측정하였다. 이때 사이토카인(cytokine) 농도의 측정 한계치는 10pg/ml이었다.It was measured by an rbent assay kit. At this time, the limit of measurement of cytokine concentration was 10 pg / ml.

비장세포 분리방법을 이용하여 C57BL/c 생쥐로부터 비장세포를 분리하여 마지막에 2ml의 10% FCS-RPMI 배지에 희석하고 200㎕의 마이토마이신 C(mytomycin C) 2mg/ml를 첨가하여 37℃ 수욕(water bath)에 25분간 처리하였다. 그 다음 세척용배지로 4℃, 1200rpm에서 6분간 원심침전을 다섯번 반복하여 세척한 다음 allogen-Splenocytes were isolated from C57BL / c mice using the splenocyte separation method, diluted in 2ml of 10% FCS-RPMI medium, and 200mg of mytomycin C was added to 2mg / ml. (water bath) was treated for 25 minutes. Then, wash repeatedly with centrifugal sedimentation for 5 minutes at 4 ℃ and 1200rpm for 5 minutes.

ic 비장세포 또는 표적세포로 사용하였다.Used as ic splenocytes or target cells.

표적세포(Target cell, allogeneic spleen cells, 1×107, 5×106/well)와 효과세포(Effetor cell,T cells, 1×106/well)의 비율(ratio)을 10:1 또는 5:1로 하여 둥근바닥 96well plate에 최종부피가 100㎕되도록 넣고 4℃, 250g에서 5분간 원심침전 시킨 후 37℃, 5% CO2배양기에 8시간 배양하였다. 단, 배양시간이 8시간이 되기 45분전에 Target maximum과 부피보정 대조구(Volume correcting control)에는 용해완충용액(Lysis Buffer)를 10배 희석하여 첨가하여 세포의 용해도가 최대가 되도록 하였다. 배양 후 4℃, 250g에서 5분간 원심침전 시킨 다음 상층액 50㎕를 떠서 평평한 바닥의 96well plate에 옮기고 여기에 50㎕ 기질혼합 완충용액(substrate mix Buffer)를 첨가하고 실온에서 알루미늄 호일을 덮어 30분간 반응시킨 다음 50㎕ 정지용액(stop soultion)을 넣고 마이크로플래이트 리더(microplate reader)를 이용하여 490nm에서 O.D.를 측정하고 상층액내에 있는 락테이트 디하이드로게나아제(lactate dehydrogenase, LDH)의 양을 측정하여 살세포작용 정도를 나타내었다. 살세포작용 정도는 하기의 식을 이용하여 산출하였다.The ratio of target cells (allogeneic spleen cells, 1 × 10 7 , 5 × 10 6 / well) and effector cells (Effetor cell, T cells, 1 × 10 6 / well) is 10: 1 or 5 : into a final volume of 96well round bottom plate 1 so as to 100㎕ were incubated 8 hours for 4 ℃, then centrifuged 5 minutes in the precipitation 250g 37 ℃, 5% CO 2 incubator. However, 45 minutes before the incubation time was 8 hours, the target maximum and volume correcting control (Lysis Buffer) was diluted 10-fold and added to the cell solubility to the maximum. After incubation, centrifuged for 5 minutes at 250 g at 4 ° C. Then, 50 µl of the supernatant was transferred to a 96-well plate on a flat bottom, and 50 µl substrate mix buffer was added thereto, and the aluminum foil was covered at room temperature for 30 minutes. After the reaction, 50 μl stop soultion was added and the OD was measured at 490 nm using a microplate reader. The amount of lactate dehydrogenase (LDH) in the supernatant was measured. The degree of killer cell activity was shown. The degree of killer cell activity was calculated using the following equation.

% CYTOTOXICITY=% CYTOTOXICITY = Expermental-Effector Spontaneous-Target SpontaneousExpermental-Effector Spontaneous-Target Spontaneous ×100× 100 Target Maximum-Target SpontaneousTarget Maximum-Target Spontaneous

생쥐의 비장세포에 택사 추출물을 농도별로 처리하여 세포의 증식정도를 측정한 결과 도 8a에 나타낸 바와 같이 무처리 대조군에 비해 택사 추출물을 처리한실험군의 비장세포 증식정도가 농도 의존적으로 증가함을 알 수 있었다. 비장세포에 포함되어 있는 B세포만을 분리하여 택사의 B세포증식 유도효과를 측정한 결과, 도 8b에 나타낸바와 같이 비장세포에서와 마찬가지로 농도 의존적으로 증식을 증가시켰으며 특히 1000㎍/㎕의 농도에서는 월등한 상승효과가 있음을 알 수 있었다.As a result of measuring the proliferation of cells by the concentration of taxa extract in the spleen cells of the mouse by concentration, as shown in FIG. 8a, the concentration of the splenocytes in the experimental group treated with the extract was increased in a concentration-dependent manner as shown in FIG. Could. As a result of measuring the induction of B cell proliferation of taxa by separating only B cells contained in splenocytes, as shown in FIG. 8B, proliferation was increased in a concentration-dependent manner as in splenocytes, especially at a concentration of 1000 ㎍ / μl. It was found that there was an excellent synergistic effect.

택사 추출물이 대식세포주의 일산화질소(NO) 생산에 미치는 효과를 조사한 결과, 표 2에 나타낸바와 같이 무처리 대조군과 비교해보면 택사 추출물을 농도별로 단독 처리한 실험군과 LPS를 1㎍/㎖을 같이 처리한 실험군에서는 모두 대식세포의 일산화탄소(NO) 생산이 유도되지 않았다. 그러나 1ng/㎖의 IFN-γ를 같이 처리한 실험군에서는 택사 추출물의 농도가 낮을때는 약간 감소하는 경향을 보였지만, 고농도에서 일산화질소(NO)생산이 유의성 있게 증가하였다. 즉, 택사추출물이 1000㎍/㎕같은 고농도에서는 IFN-γ에 의해 유도되는 일산화질소(NO) 생산에 상승적 작용을 하는 것을 알 수 있었다.As a result of investigating the effect of taxa extract on nitric oxide (NO) production of macrophage, as compared to the untreated control group, as shown in Table 2, the experimental group treated with taxa extract alone by concentration and LPS were treated together with LPS. None of the experimental groups induced carbon monoxide (NO) production in macrophages. However, the experimental group treated with 1 ng / ml IFN-γ showed a tendency to decrease slightly at low concentrations of taxa extract, but significantly increased NO production at high concentrations. In other words, it can be seen that the taxa extract has a synergistic effect on the production of nitrogen monoxide (NO) induced by IFN-γ at high concentrations such as 1000 µg / µl.

택사 추출물이 대식세포주 RAW 264.7의 일산화질소 생산에 미치는 영향Effects of Taxa Extracts on Nitric Oxide Production in Macrophage Cell RAW 264.7 택사 추출물Taxa Extract 자 극 제Stimulant 00 LPS(1㎍/㎖)LPS (1 μg / ml) IFN-γ(1ng/㎖)IFN-γ (1 ng / ml) 00 4.17±1.574.17 ± 1.57 3.81±1.253.81 ± 1.25 5.47±1.745.47 ± 1.74 1One 2.90±0.132.90 ± 0.13 3.12±0.083.12 ± 0.08 2.86±0.232.86 ± 0.23 1010 2.87±0.242.87 ± 0.24 2.66±0.192.66 ± 0.19 3.50±0.203.50 ± 0.20 100100 3.02±0.103.02 ± 0.10 3.02±0.713.02 ± 0.71 6.43±0.016.43 ± 0.01 10001000 4.49±1.474.49 ± 1.47 5.87±2.625.87 ± 2.62 21.44±3.0421.44 ± 3.04 〔주〕택사 추출물 단위: ㎍/㎖Taxa extract unit: µg / ml

택사 추출물이 동종항원에 의한 증식반응에 미치는 효과를 조사한 결과, 도9a에 나타낸 바와 같이 동일한 표적세포의 수(5×105cells/well)일 때 효과세포의 수가 well당 1×106cells이 배양 후 3일째에 최대 증식반응을 나타내었다. 따라서, 동일한 조건에서 0.01, 0.1, 1, 10㎍/㎖의 택사 추출물을 같이 처리하여 배양 후 3일째 증식반응을 측정한 결과, 도 9b에 나타낸 바와 같이 택사 추출물의 농도 의존적으로 동종항원에 대한 증식반응이 억제되었다. 즉, 1㎍/㎖에서 최대 약 50%정도의 증식 억제효과를 나타냈다.As a result of investigating the effect of taxa extract on the proliferative response by homologous antigen, as shown in FIG. 9A, when the same number of target cells (5 × 10 5 cells / well), the number of effect cells was 1 × 10 6 cells per well. Maximum proliferative response was seen at 3 days after culture. Therefore, as a result of measuring the proliferative response on day 3 after cultivation by treating 0.01, 0.1, 1, 10 ㎍ / ml taxa extract under the same conditions, as shown in FIG. The reaction was suppressed. In other words, the maximum inhibitory effect of about 50% at 1 ㎍ / ㎖.

T세포가 생산하는 사이토카인(cytokine)의 생산량을 측정한 결과, 표 3에 나타낸 바와 같이 동종항원에 대해 반응하는 T세포는 IL-2, IL-10 및 IFN-γ 등을 생산하였지만, 택사 추출물을 첨가한 실험군에서 IL-2, IFN-γ 및 IL-10의 생산량이 억제됨을 알 수 있었다. 따라서, 택사 추출물은 동종항원에 대해 반응하는 T세포의 사이토카인(cytokine) 생산을 억제함으로써 분열 증식을 저해하는 것으로 생각된다.As a result of measuring the amount of cytokines produced by T cells, T cells responding to homologous antigens produced IL-2, IL-10 and IFN-γ, as shown in Table 3. It was found that the production of IL-2, IFN-γ and IL-10 was suppressed in the experimental group added. Therefore, it is thought that the taxa extract inhibits cleavage proliferation by inhibiting cytokine production of T cells in response to homologous antigens.

택사 추출물이 동종항원에 대해 반응하는 T세포의 사이토카인 분비에 미치는 영향Effects of Taxa Extracts on Cytokine Secretion of T Cells Responding to Homologous Antigens 실험 조건Experimental conditions 사이토카인(cytokines)Cytokines IL-2IL-2 IFN-γIFN-γ IL-4IL-4 IL-10IL-10 T 세포T cell <10<10 94.3±17.294.3 ± 17.2 <10<10 200.0±21.2200.0 ± 21.2 T 세포+ 비장T cell + spleen 372.4±4.2372.4 ± 4.2 644.3±12.1644.3 ± 12.1 104±13.3104 ± 13.3 430.0±148.5430.0 ± 148.5 T 세포+비장+택사 추출물T Cell + Spleen + Taxa Extract 316.4±3.7316.4 ± 3.7 422.9±23.2422.9 ± 23.2 <10<10 205.0±70.7205.0 ± 70.7 〔주〕사이토카인 단위: pg/㎖(Note) Cytokine Unit: pg / ml

택사 추출물이 동종항원에 대한 T세포의 살세포작용에 미치는 영향을 조사한결과, 도 10에 나타낸바와 같이 동종항원에 대한 T세포의 살세포 작용을 택사 추출물이 저농도인 1㎍/㎖에서 완전히 억제함을 알 수 있었다. 따라서, 택사 추출물은 동종항원에 대한 T세포의 사이토카인(cytokine) 생산량을 억제함으로써 분열증식을 억제하였고, T세포의 동종항원에 대한 살세포 작용도 억제하는 것으로 나타났다.As a result of investigating the effect of taxa extract on T cell apoptosis on allogeneic antigen, as shown in FIG. 10, T. agar extract completely inhibited T cell apoptosis on homologous antigen at low concentration of 1㎍ / ml And it was found. Therefore, the extract of Taeksa inhibited the proliferation by inhibiting the cytokine production of T cells against homologous antigens, and also inhibited the killer cell activity against homologous antigens of T cells.

실험예 4: 택사복음차의 복음처리조건의 결정Experimental Example 4: Determination of Gospel Processing Conditions of Taxa Gospel

택사는 복음기(roaster)를 사용하여 복음온도 130, 140, 150, 160 및 170℃에서 30분으로 설정하고 복음처리하였다. 복음처리한 치커리를 Laboratory Mill로 20∼45mesh 크기로 분쇄한 다음 복음택사분말의 관능평가를 실시하고 색도의 변화 및 택사물추출물의 특성을 조사하였다. 관능평가는 9점 기호척도법으로 평가하였으며 색도의 변화는 훈터색도(Hunter color values)를 사용하였다. 택사 물추출물의 특성은 복음처리한 택사 20g에 증류수 200ml을 첨가하여 80℃에서 5시간 추출하여 여과한 뒤 200ml로 정용하여 복음처리 조건에 따른 pH의 변화, 갈색도 및 탁도를 측정하였다.Taxi was set to 30 minutes at gospel temperatures 130, 140, 150, 160 and 170 ° C. using a roaster, and the gospel was processed. Gospel treated chicory was pulverized to 20 ~ 45mesh in Laboratory Mill, and then sensory evaluation of Gospel-Taxi powder was carried out to investigate the change of chromaticity and the characteristics of the extract. Sensory evaluation was evaluated by 9-point symbol scale method, and Hunter color values were used for the change of chromaticity. The characteristics of the water extracts of taxi water were added 200ml of distilled water to 20g of the gospel treated texas, extracted at 80 ° C. for 5 hours, filtered, and then applied to 200ml to measure the pH change, brownness and turbidity according to the gospel treatment conditions.

실험 결과, 표 4에 나타낸 바와 같이 복음처리하지 않은 무처리구에서는 마른 풀냄새가 7.4로 높게 나타났으며 고소한 냄새(누룽지 냄새), 단냄새 등은 낮게 나타나 전반적인 기호성이 낮게 나타났다. 복음온도가 증가하고 복음시간이 길수록 고소한 냄새(누룽지 냄새), 단냄새, 종합적 기호도 등이 증가하는 경향으로 나타났으며, 고소한 냄새는 160℃, 30분 복음처리했을 때 7.4, 단냄새는 고소한 냄새와 마찬가지로 160℃, 30분에서 6.9였다. 마른 풀냄새는 복음온도가 높아지고 복음시간이 길어질수록 뚜렷이 감소하였으며 탄냄새는 복음온도가 높아지고 복음시간이 길수록 급속하게 증가하는 경향으로 나타났다. 특히 160℃ 30분, 40분에서는 각각 7.7과 8.1 으로 기호성이 매우 나쁘게 나타났다. 종합적 기호도는 160℃에서 7.1점으로 가장 우수하게 나타났다.As a result, as shown in Table 4, the dry grass odor was high as 7.4 in untreated vines, and the odor (Nurungji odor) and sweet odor were low, resulting in low overall palatability. As the Gospel temperature increased and the Gospel time increased, the odor (Nurungji odor), sweet odor, and overall taste tended to increase. Similarly, it was 6.9 at 160 ° C for 30 minutes. Dry grass odor significantly decreased with increasing gospel temperature and longer gospel time, and burnt smell tended to increase rapidly with higher gospel temperature and longer gospel time. In particular, at 160 ℃ 30 minutes and 40 minutes, palatability was very bad as 7.7 and 8.1, respectively. The overall acceptability was the best at 7.1 points at 160 ℃.

복음조건에 따른 택사복음차의 관능평가 결과Sensory Evaluation Results of the Gospel Differences According to Gospel Conditions 복음조건Gospel Condition 관능적 특성Sensory characteristics 온도(℃)Temperature (℃) 시간(분)Minutes 구운쌀냄새Grilled Rice Smell 단 냄새Sweet smell 마른풀냄새Smell of dry grass 탄 냄새Burnt smell 전체기호도Overall Symbol 무처리구No treatment 1.01.0 2.72.7 7.47.4 1.01.0 2.02.0 130130 3030 4.34.3 4.14.1 3.23.2 2.62.6 4.84.8 140140 3030 5.25.2 5.45.4 2.52.5 3.33.3 5.25.2 150150 3030 6.16.1 6.86.8 2.72.7 4.04.0 6.66.6 160160 3030 7.47.4 6.96.9 2.02.0 4.94.9 7.17.1 170170 3030 5.85.8 4.64.6 1.21.2 7.77.7 3.53.5

복음처리한 택사 분말의 색도변화는 표 5에 나타낸 바와 같다. 즉, 복음온도가 증가 할수록 명도를 나타내는 L 값은 무처리구의 경우 66.62였으며 170℃에서는 55.72로 어둡게 나타났다. 붉은색(+a)에서 녹색(-a)을 나타내는 a값은 무처리구에서는 -3.36으로 나타났으며 복음온도 170℃에서는 5.83으로 붉은색으로 나타났다. 황색(+b)에서 청색(-b)을 나타내는 b값은 무처리구의 경우 17.11이였으며 복음 온도 170℃에서는 36.14의 황색으로 변하였다.The chromaticity change of the gospel-treated tackifier powder is shown in Table 5. That is, as Gospel temperature increased, the L-value indicating brightness was 66.62 in the untreated zone and darkened to 55.72 at 170 ℃. The value of a representing red (+ a) to green (-a) was -3.36 in the untreated group and 5.83 at the Gospel temperature of 170 ℃. The b-value from blue (-b) to blue (-b) was 17.11 for the untreated group and changed to yellow at 36.14 at the gospel temperature of 170 ° C.

총색차(ΔE)는 볶음온도가 증가할수록 크게 변하는 것으로 나타났다.The total color difference (ΔE) was found to change significantly with increasing roasting temperature.

택사의 복음조건에 따른 훈터색도(Hunter color values)의 변화Changes in Hunter Color Values According to Gospel Terms of Taxi 복음 조건Gospel conditions 훈터 색도(Hunter's color values)Hunter's color values 온도(℃)Temperature (℃) 시간(분)Minutes LL aa bb ΔEΔE 무처리구No treatment 66.5266.52 -3.36-3.36 17.1117.11 34.6734.67 130130 3030 66.8066.80 -0.02-0.02 23.7823.78 37.3837.38 140140 3030 71.7771.77 0.020.02 29.0529.05 37.0137.01 150150 3030 68.1868.18 1.651.65 30.5030.50 40.7040.70 160160 3030 62.2462.24 3.203.20 31.3431.34 46.0046.00 170170 3030 55.7255.72 5.835.83 30.1430.14 50.6850.68

복음처리한 택사의 물추출물의 특성은 표 6에 나타낸 바와 같다. pH는 무처리구의 경우 6.66으로 나타났으며 복음온도가 증가할수록 낮아져 170℃에서는 5.57이였다. 갈색도는 무처리구에서는 0.313로 나타났으며 복음온도가 증가할수록 높아져 170℃에서는 0.481로 나타났다. 탁도는 무처리구에서는 66.71이였으며 복음온도가 증가할수록 탁해져 170℃에서는 89.86으로 탁하게 나타났다. 따라서 택사복음차의 최적 복음조건을 160℃, 30분간으로 선정하였다.The characteristics of the water extracts of the tackified taxa are shown in Table 6. The pH was 6.66 in the untreated and lowered with increasing Gospel temperature, which was 5.57 at 170 ℃. Brownness was 0.313 in the untreated area and increased with increasing Gospel temperature, 0.481 at 170 ℃. Turbidity was 66.71 in the untreated zone, and it became turbid as the Gospel temperature increased to 89.86 at 170 ℃. Therefore, the optimal gospel condition of the Gospel of Texas was selected as 160 ℃ for 30 minutes.

복음처리한 택사의 물추출물의 특성Characteristics of Water Extracts of Gospel Treated Taxi 복음 조건Gospel conditions pHpH 갈색도(420nm)Brownness (420 nm) 탁도(660nm)Turbidity (660 nm) 온도(℃)Temperature (℃) 시간(분)Minutes 무처리군No treatment group 6.666.66 0.3130.313 67.2167.21 130130 3030 6.606.60 0.3230.323 74.5374.53 140140 3030 5.705.70 0.3430.343 78.0478.04 150150 3030 5.615.61 0.4050.405 80.8380.83 160160 3030 5.435.43 0.4000.400 87.3787.37 170170 3030 5.375.37 0.4810.481 89.8689.86

실시예 1: 택사복음차의 제조Example 1 Preparation of Taxa Gospel Tea

제 1공정: 택사의 복음처리Process 1: Gospel Processing of Taxi

택사의 기호성과 기능성을 증진시키기 위하여 택사를 복음기를 사용하여 실험예 4에서 결정된 최적조건인 복음온도 160℃에서 30분간 복음처리하였다. 즉, 예열된 복음기에 건조시킨 건조택사 100g을 넣고 20rpm 속도로 드럼을 회전시키면서 복음처리하였다. 이때 복음온도의 편차는 ±1℃이내였다.In order to enhance the palatability and functionality of the taxi, the taxi was evangelized for 30 minutes at a gospel temperature of 160 ° C., which is the optimum condition determined in Experimental Example 4 using the Gospel. That is, 100 g of the dried taxi tackled in the preheated Gospel was put and the gospel was processed while rotating the drum at a speed of 20 rpm. At this time, the deviation of the gospel temperature was within ± 1 ℃.

제 2공정: 일반음용형 택사복음차의 제조Second Process: Preparation of General Drinking Taxa Gospel

제 1공정에서 복음처리한 택사를 알류미늄호일 티백(Al-foil)에 10g씩 포장하여 제조하였다.The texas treated in the first step was prepared by packing 10g each in aluminum foil tea bags (Al-foil).

제 3공정: 택사복음차의 분쇄 및 포장Process 3: Crushing and Packing the Taxi Gospel

제 1공정에서 복음처리한 택사를 실험실용 분쇄기(Laboratory Mill)로 20∼45mesh 크기로 분쇄하여 약 1.4g씩 부직포에 티백(Tea bag)용으로 포장하여 제조하였다.The tackifier treated in the first step was pulverized to a size of 20 to 45 mesh using a laboratory mill, and then packed in a nonwoven fabric by about 1.4 g for a tea bag.

이상, 상기 실시예를 통하여 설명한 바와 같이 본 발명의 항균활성, 항산화효과 및 면역활성이 뛰어난 택사를 건조시킨 뒤 복음기를 이용하여 복음처리한 후 포장하여 일반음용형 차로 제조하거나 분쇄한 다음 티백(Tea bag)으로 포장하여 침출형 차를 제조함으로서 기호성이 증진된 택사복음차를 제조하는 효과가 있으므로 식음료산업과 국민건강증진에 있어서 매우 유용한 발명인 것이다.As described above, as described through the above examples, the antimicrobial, antioxidative and immune activities of the present invention are dried, then tackified using Gospel, then packaged and manufactured or pulverized as a general drinking tea after tea bag (Tea) It is a very useful invention in the food and beverage industry and the public health because it has the effect of manufacturing the tactical Gospel tea with improved palatability by manufacturing a leaching type tea by packaging in a bag).

Claims (4)

(a) 택사 100g당 150ml의 비율로 에탄올을 첨가하고 80℃에서 1 내지 3 시간 동안 추출하는 단계;(a) adding ethanol at a rate of 150 ml per 100 g of taxi and extracting at 80 ° C. for 1 to 3 hours; (b) 상기 추출된 택사를 건조시키는 단계;(b) drying the extracted taxi; (c) 상기 건조된 택사를 복음기에 넣고 20rpm 속도로 회전시키면서 150 내지 160℃에서 30분 내지 60분간 복음처리하는 단계;(c) putting the dried tack in the gospel to the gospel process while rotating at 20 rpm speed for 30 to 60 minutes at 150 to 160 ℃; (d) 상기 (c)단계의 복음처리한 택사를 Al-foil 포장하여 일반음용형 복음차로 제조하는 단계를 포함하는 것을 특징으로 하는 택사복음차 제조방법.(D) a method for producing a tack gospel gospel comprising the step of producing a general drink type Gospel by packaging Al-foil of the tackified tack in step (c). (a) 택사 100g 당 1500ml의 비율로 에탄올을 첨가하고 80℃에서 1 내지 3시간 동안 택사를 추출하는 단계;(a) adding ethanol at a rate of 1500 ml per 100 g of taxa and extracting the taxa at 80 ° C. for 1 to 3 hours; (b) 상기 추출된 택사를 건조시키는 단계;(b) drying the extracted taxi; (c) 상기 건조된 택사를 복음기에 넣고 20rpm 속도로 회전시키면서 150 내지 160℃에서 30분 내지 60분간 복음처리하는 단계;(c) putting the dried tack in the gospel to the gospel process while rotating at 20 rpm speed for 30 to 60 minutes at 150 to 160 ℃; (d) 상기 (c)단계의 복음처리한 택사를 분쇄기를 사용하여 20 내지 45mesh 크기로 분쇄하고 티백(tea bag)에 포장하여 침출형 차로 제조하는 단계로 구성되는 것을 특징으로 하는 택사복음차 제조방법.(d) preparing a gospel Gospel tea comprising the step of pulverizing the tackified texas of step (c) using a grinder to a size of 20 to 45mesh and packing it into a tea bag to produce a leachable tea Way. 제 1항의 방법에 의해 제조된 택사복음차.Taxi gospel produced by the method of claim 1. 제 2항의 방법에 의해 제조된 택사복음차.Taxi gospel produced by the method of claim 2.
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KR19980016185A (en) * 1996-08-27 1998-05-25 김광수 Manufacturing method of chrysanthemum tea
KR19980044841A (en) * 1996-12-09 1998-09-15 임희택 Manufacturing method of low fat herbal tea
KR100249281B1 (en) * 1998-02-19 2000-03-15 김홍엽 The method process of manufacture of chinese matrimony vine tea
KR20000018127A (en) * 2000-01-13 2000-04-06 김종성 The manufacturing for method and apparatus of mulberry tree tea
KR20010081369A (en) * 2000-02-14 2001-08-29 허경만 Slimming green tea and method for preparation thereof

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR19980016185A (en) * 1996-08-27 1998-05-25 김광수 Manufacturing method of chrysanthemum tea
KR19980044841A (en) * 1996-12-09 1998-09-15 임희택 Manufacturing method of low fat herbal tea
KR100249281B1 (en) * 1998-02-19 2000-03-15 김홍엽 The method process of manufacture of chinese matrimony vine tea
KR20000018127A (en) * 2000-01-13 2000-04-06 김종성 The manufacturing for method and apparatus of mulberry tree tea
KR20010081369A (en) * 2000-02-14 2001-08-29 허경만 Slimming green tea and method for preparation thereof

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