KR20220043008A - Antioxidant or Anti-bacterial Composition Using an Extract of Ludwigia epilobioides - Google Patents
Antioxidant or Anti-bacterial Composition Using an Extract of Ludwigia epilobioides Download PDFInfo
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Abstract
Description
본 발명은 여뀌바늘(Ludwigia epilobioides) 추출물을 이용한 항산화 또는 항균용 조성물에 관한 것이다.The present invention relates to an antioxidant or antibacterial composition using an extract of Ludwigia epilobioides .
생체 내 정상적인 세포대사과정에서 생성되거나 여러 환경오염 및 화학물질의 노출 등에 의해 생성되는 과산화 라디칼(superoxide radical, O2ㆍ), 수산화 라디칼(hydroxyl radical, OHㆍ), 과산화수소(hydrogen peroxide, H2O2), 단일체 산소(singlet oxygen, 1O2) 등을 총칭하여 활성산소종(ROS: reactive oxygen species)이라고 한다.Superoxide radical (O 2 ㆍ), hydroxyl radical (OH ㆍ), hydrogen peroxide (H 2 O 2 ), single oxygen ( 1 O 2 ), and the like are collectively called reactive oxygen species (ROS).
이들 활성 산소는 반응성 매우 커서 생체 분자들과 빠르게 반응하여 생체막의 지질 산화, 단백질 변성, DNA 손상 등을 초래하며, 암, 동맥경화증, 당뇨병, 자가면역질환, 노화, 뇌졸중, 파킨슨병 등 각종 질환의 발병 위험성을 증가시키는 것으로 알려져 있다(Chem. Res. 14: 393 (1981); Arch. Biophys. 247: 1-11 (1986); Science 221: 1256-1264 (1983)).These reactive oxygen species are very reactive and react rapidly with biomolecules, causing lipid oxidation, protein denaturation, DNA damage, etc. of biological membranes. It is known to increase the risk of developing disease ( Chem. Res. 14: 393 (1981); Arch. Biophys. 247: 1-11 (1986); Science 221: 1256-1264 (1983)).
생명체는 지속적으로 발생하는 이러한 산화적 스트레스에 대응하기 위한 항산화 체계를 갖추고 있는데, 이러한 항산화 체계는 크게 슈퍼옥사이드 디스뮤타아제, 카탈라아제, 글루타티온 퍼옥시다아제, 글루타티온 환원효소, 글루타티온 S-전이효소 등에 의한 효소적 체계와 식이에 의하여 섭취되는 비타민 A, 비타민 C, 비타민 E, 항산화 무기질 등의 항산화제에 의한 비효소적 체계로 구분된다(Annu. Rev. Nutr., 16, pp33-50 (1996)). 효소적 체계와 비효소적 체계는 상호 보완적으로 작용하여 생체에서 생성되는 산화성 물질을 무독화시킨다.Organisms have an antioxidant system to respond to this continuously occurring oxidative stress, and these antioxidant systems are largely enzymatic by superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, etc. It is divided into a non-enzymatic system by antioxidants such as vitamin A, vitamin C, vitamin E, and antioxidant minerals ingested through the system and diet ( Annu. Rev. Nutr. , 16, pp33-50 (1996)). The enzymatic system and the non-enzymatic system work complementary to each other to detoxify oxidative substances produced in the body.
항산화제는 활성 산소와 반응함으로써 활성 산소에 의한 생체 손상을 억제하거나 최소화하는 것으로, 부틸화 하이드록시아니솔(BHA, Butylated hydroxyanisole), 부틸화 하이드록시톨루엔(BHT, Butylated hydroxytoluene), 프로필 갈레이트(PG, Propyl galate), 터셔리부틸 하이드로퀴논(TBHQ, Teritiarybutyl hydroquinone) 등의 합성 항산화제와 엽록소, 녹차 추출물, 프로롤리스 추출물, 클로렐라, 비타민 E, 비타민 C, 스쿠알레, 플라보노이드, 대나무 잎 추출물 등의 천연 항산화제로 구분될 수 있는데, 합성 항산화제의 경우 발암 및 심혈관 계열에 부작용을 유발하는 것으로 알려지면서 보다 안전하고 뛰어난 효과를 가지는 식물 유래 항산화제에 관한 연구가 활발하게 수행되고 있다(Korean J. Food Sci. Technol. 34:889-892 (2002)).Antioxidants inhibit or minimize damage to the living body by reactive oxygen species by reacting with active oxygen. Synthetic antioxidants such as PG, Propyl galate) and tertiarybutyl hydroquinone (TBHQ, Teritiarybutyl hydroquinone) and It can be classified as a natural antioxidant, and as synthetic antioxidants are known to cause side effects on carcinogenesis and cardiovascular systems, research on plant-derived antioxidants that are safer and have excellent effects are being actively conducted ( Korean J. Food ). Sci. Technol. 34:889-892 (2002)).
식물은 항산화성 물질인 폴리페놀, 플라보노이드 등을 함유하고 있어 대부분의 식물 추출물은 항산화 활성을 나타내지만, 식물 유래의 항산화제는 항산화 효과가 일반적으로 합성 항산화제에 비하여 매우 낮기 때문에 실효성이 있는 항산화 효과를 나타내기 위해서는 다량으로 사용하여야 하고 결국 원료 가격의 상승으로 경제성과 실용성이 떨어지는 문제점이 있다. 이러한 이유로 식물 유래 항산화제의 발굴은 합성 항산화제에 버금가는 활성을 나타내는 물질을 찾는데 집중되어 있다.Plants contain antioxidant substances such as polyphenols and flavonoids, so most plant extracts show antioxidant activity. In order to show the , it must be used in large quantities, and consequently, there is a problem in that economical efficiency and practicality are deteriorated due to an increase in the price of raw materials. For this reason, the excavation of plant-derived antioxidants is focused on finding substances with activity comparable to synthetic antioxidants.
최근 항산화 활성을 보이는 물질을 탐색하기 위한 항산화 활성 측정 기법으로서 TRAP 법(Anal Biochem., 28, pp300-306. 1989), DPPH 법(Nature 181:1198-1200 (1958)), ABTS 법(Methods Enzymol. 234: 279-293 (1994); Clin. Sci. 84: 407-412 (1993); Free Rad. Biol. Med. 20: 933-956 (1996)), ORAC 법(Free Rad. Biol. Med. 18:29-36 (1995)), TOSC 법(Free Rad. Biol. Med. 24(3):480-493 (1998); Toxicol. Appl. Pharmacol. 156(2):96-105 (1999)), Potassium ferricyanide 법(Jpn, J. of Nutr. 44:307-315, 1986), SOD 유사활성 측정법(Eur J Biochem 47:469-474, 1974) 등 다양한 방법이 개발되어 있다.TRAP method ( Anal Biochem. , 28, pp300-306. 1989), DPPH method ( Nature 181:1198-1200 (1958)), ABTS method ( Methods Enzymol ) 234 : 279-293 (1994); Clin. Sci. 84: 407-412 (1993); Free Rad. Biol. Med. 20: 933-956 (1996)), ORAC method ( Free Rad. Biol. Med. 18:29-36 (1995)), TOSC method ( Free Rad. Biol. Med. 24(3):480-493 (1998); Toxicol. Appl. Pharmacol. 156(2):96-105 (1999)) , Potassium ferricyanide method (Jpn, J. of Nutr. 44:307-315, 1986), SOD-like activity measurement method ( Eur J Biochem 47:469-474, 1974), such as various methods have been developed.
한편 세균의 감염은 인간의 질병에서 가장 흔하고 치명적인 원인 중의 하나이다. 특히 병원성 미생물에 의한 직접 혹은 간접적인 피해는 경제, 환경, 의학적으로 많은 문제를 야기시키고 있다. 병원성 미생물은 식품 산업에서의 식품 유통 과정 중의 부패로 인한 손실, 농산업에서의 농작물에 대한 과량의 화학 살충제의 사용으로 인한 인체의 유해성 및 환경오염, 항생제의 오남용으로 인한 항생제 내성 균주의 출현 등 사회 전반에서 많은 문제들을 야기시키고 있다Meanwhile, bacterial infection is one of the most common and fatal causes of human diseases. In particular, direct or indirect damage caused by pathogenic microorganisms causes many economic, environmental and medical problems. Pathogenic microorganisms are harmful to the human body and environmental pollution due to the use of excessive chemical pesticides on crops in the agricultural industry, the loss due to spoilage during the food distribution process in the food industry, and the emergence of antibiotic-resistant strains due to the misuse of antibiotics. is causing many problems in
특히 황색포도상구균(Staphylococcus aureus)은 통성 혐기성(facultatively anaerobic)의 그람 양성 구균으로써 포도상구균 감염증의 가장 흔한 원인균이다. 포도상구균은 건강한 사람의 피부와 비강의 점막에서 집락을 형성하며 높은 보균율로 인하여 인체에 매우 흔한 감염증을 일으킨다. 포도상구균에 의한 감염증에는 여드름, 농가진(膿痂疹), 창상과 같은 피부감염증에서부터 폐렴, 패혈증, 심내막염 등의 생명을 위협하는 중증 질환에 이르기까지 다양한 질병을 초래할 수 있다(J Heart Lung Transplant 2012, 31(11):1199-1206; Microb Pathog 2013;54:26-33; Eur J Clin Microbiol Infect Dis 1994;13(1):50-55).In particular, Staphylococcus aureus is a facultatively anaerobic Gram-positive cocci and is the most common causative agent of staphylococcal infections. Staphylococcus aureus forms colonies on the skin and nasal mucosa of healthy people and causes very common infections in the human body due to its high carrier rate. Staphylococcus aureus infection can cause a variety of diseases, ranging from skin infections such as acne, impetigo, and cuts to serious, life-threatening diseases such as pneumonia, sepsis, and endocarditis (J Heart Lung Transplant 2012, 31(11):1199-1206; Microb Pathog 2013;54:26-33; Eur J Clin Microbiol Infect Dis 1994;13(1):50-55).
이러한 황색포도상구균 중에서 페니실린계 항생제인 메씨실린(methicillin)에 내성을 보이는 균을 메티실린 내성 황색포도상구균(methicillin-resistant Staphylococcus aureus, MRSA)이라고 하며, 1961년 영국에서 처음 출현하여 1980년대 이후 급격하게 증가하고 있는데, 최근에는 우리나라 종합병원이나 대학병원에서 MRSA가 황색포도상구균 균주 중 70% 이상 차지하는 것으로 알려져 있다. 모든 베타-락탐(β-lactam)계 항생제에 내성이 있으며 또 귀놀론(quinolone)계 항생제를 비롯한 대부분의 항생제에도 강한 내성을 가지고 있어 감염 후 사망률이 높기 때문에 전 세계적으로 심각한 문제로 대두되고 있다(N Engl J Med 2005, 352:1436-44; Clin Infect Dis 1994, 19(6):1123-8).Among these Staphylococcus aureus, bacteria that are resistant to methicillin, a penicillin-based antibiotic, are called methicillin-resistant Staphylococcus aureus ( MRSA). Recently, it is known that MRSA accounts for more than 70% of Staphylococcus aureus strains in general hospitals or university hospitals in Korea. It is resistant to all β -lactam antibiotics and also has strong resistance to most antibiotics including quinolone antibiotics. N Engl J Med 2005, 352:1436-44; Clin Infect Dis 1994, 19(6):1123-8).
본 발명은 여뀌바늘 추출물의 항산화 활성 및 항균 활성을 개시한다.The present invention discloses antioxidant and antibacterial activity of extract
본 발명의 목적은 여뀌바늘 추출물을 이용한 항산화 또는 항균용 조성물을 제공하는 데 있다.It is an object of the present invention to provide an antioxidant or antibacterial composition using the extract
본 발명의 다른 목적이나 구체적인 목적은 이하에서 제시될 것이다.Other objects or specific objects of the present invention will be set forth below.
본 발명은 아래의 실시예 및 실험예에서 확인되는 바와 같이, 여뀌바늘 추출물이 DPPH 법, 환원력 측정(Reducing power)에 따른 항산화 활성을 가지고, 황색포도상구균(Staphylococcus aureus), 메티실린 내성 황색포도상구균(Methicillin-resistant Staphylococcus aureus, MRSA), 및 대장균에 대해 항균 활성을 가짐을 확인함으로써 완성된 것이다.As confirmed in the Examples and Experimental Examples below, the present invention has antioxidant activity according to the DPPH method and reducing power, Staphylococcus aureus , methicillin-resistant Staphylococcus aureus. (Methicillin-resistant Staphylococcus aureus , MRSA), and was completed by confirming that it has antibacterial activity against E. coli.
전술한 바를 고려할 때, 본 발명은 여뀌바늘 추출물을 유효성분으로 포함하는 항산화용 조성물 또는 항균용 조성물로 파악할 수 있다.Considering the above, the present invention can be identified as an anti-oxidation composition or antibacterial composition comprising a genus oleifera extract as an active ingredient.
본 명세서에서, "여뀌바늘 추출물"이란 추출 대상인 여뀌바늘의 줄기, 잎, 열매, 꽃, 뿌리, 지하부, 지상부 또는 이들의 혼합물을 물, 탄소수 1 내지 4의 저급 알콜(메탄올, 에탄올, 부탄올 등), 메틸렌클로라이드, 에틸렌, 아세톤, 헥산, 에테르, 클로로포름, 에틸아세테이트, 부틸아세테이트, N,N-디메틸포름아미드(DMF), 디메틸설폭사이드(DMSO), 1,3-부틸렌글리콜, 프로필렌글리콜 또는 이들의 혼합 용매를 사용하여 침출하여 얻어진 추출물, 이산화탄소, 펜탄 등 초임계 추출 용매를 사용하여 얻어진 추출물 또는 그 추출물을 분획하여 얻어진 분획물을 의미하며, 추출 방법은 활성물질의 극성, 추출 정도, 보존 정도를 고려하여 냉침, 환류, 가온, 초음파 방사, 초임계 추출 등 임의의 방법을 적용할 수 있다. 분획된 추출물의 경우 추출물을 특정 용매에 현탁시킨 후 극성이 다른 용매와 혼합·정치시켜 얻은 분획물, 상기 조추출물을 실리카겔 등이 충진된 칼럼에 흡착시킨 후 소수성 용매, 친수성 용매 또는 이들의 혼합 용매를 이동상으로 하여 얻은 분획물을 포함하는 의미이다. 또한 상기 추출물의 의미에는 동결건조, 진공건조, 열풍건조, 분무건조 등의 방식으로 추출 용매가 제거된 농축된 액상의 추출물 또는 고형상의 추출물이 포함된다. 바람직하게는 추출용매로서 물, 에탄올 또는 이들의 혼합 용매를 사용하여 얻어진 추출물, 더 바람직하게는 추출용매로서 물과 에탄올의 혼합 용매를 사용하여 얻어진 추출물을 의미한다.In the present specification, the term "extract of chrysanthemum extract" refers to extracting the stem, leaf, fruit, flower, root, underground part, above-ground part, or mixtures thereof with water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.) , methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or these It means an extract obtained by leaching using a mixed solvent of In consideration of this, any method such as chilling, reflux, warming, ultrasonic radiation, or supercritical extraction may be applied. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the crude extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by methods such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. Preferably, it refers to an extract obtained by using water, ethanol, or a mixed solvent thereof as an extraction solvent, and more preferably an extract obtained by using a mixed solvent of water and ethanol as an extraction solvent.
또 본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In addition, as used herein, the term "active ingredient" refers to a component that alone exhibits the desired activity or can exhibit activity together with a carrier that has no activity by itself.
본 발명의 항산화 또는 항균용 조성물(이하 "본 발명의 조성물")은 그 유효성분을 용도, 제형, 배합 목적 등에 따라 치료를 의도하는 알러지 질환의 개선 활성 등을 나타낼 수 있는 한 임의의 양(유효량)으로 포함할 수 있는데, 통상적인 유효량은 조성물 전체 중량을 기준으로 할 때 0.001 중량 % 내지 15 중량 % 범위 내에서 결정될 것이다. 여기서 "유효량"이란 그 적용 대상인 포유동물 바람직하게는 사람에게 의료 전문가 등의 제언에 의한 투여 기간 동안 본 발명의 조성물이 투여될 때, 항산화, 항균 효과 등 의도한 의료적·약리학적 효과를 나타낼 수 있는, 본 발명의 조성물에 포함되는 유효성분의 양을 말한다. 이러한 유효량은 당업자의 통상의 능력 범위 내에서 실험적으로 결정될 수 있다.Antioxidant or antibacterial composition of the present invention (hereinafter "composition of the present invention") can be used in any amount (effective amount) ), and a typical effective amount will be determined within the range of 0.001 wt % to 15 wt % based on the total weight of the composition. Herein, the term "effective amount" refers to the intended medical and pharmacological effects, such as antioxidant and antibacterial effects, when the composition of the present invention is administered to a mammal, preferably a human, to which it is applied during the administration period as suggested by a medical professional. It refers to the amount of the active ingredient included in the composition of the present invention. Such an effective amount can be determined empirically within the ordinary ability of one of ordinary skill in the art.
본 발명의 조성물은 구체적인 양태에 있어서, 식품 조성물로서 파악할 수 있다. 본 발명의 조성물이 식품 조성물로 파악될 경우 그 용도는 알러지성 피부 과민 반응 억제로 이해될 수 있다.In a specific aspect, the composition of this invention can be grasped|ascertained as a food composition. When the composition of the present invention is identified as a food composition, its use may be understood to inhibit allergic skin hypersensitivity reaction.
본 발명의 식품 조성물은 어떠한 형태로도 제조될 수 있으며, 예컨대 차, 쥬스, 탄산음료, 이온음료 등의 음료류, 우유, 요구르트 등의 가공 유류, 껌류, 떡, 한과, 빵, 과자, 면 등의 식품류, 정제, 캡슐, 환, 과립, 액상, 분말, 편상, 페이스트상, 시럽, 겔, 젤리, 바 등의 건강기능식품 제제류 등으로 제조될 수 있다. The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated drinks, and ion drinks, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, bread, confectionery, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and health functional food preparations such as bars can be manufactured.
또 본 발명의 식품 조성물은 법률상·기능상의 구분에 있어서 제조·유통 시점의 시행 법규에 부합하는 한 임의의 제품 구분을 띨 수 있다. 예컨대 한국 「건강기능식품에관한법률」에 따른 건강기능식품이거나, 한국 「식품위생법」의 식품공전(식약처 고시 「식품의 기준 및 규격」)상 각 식품유형에 따른 과자류, 두류, 다류, 음료류, 특수용도식품 등일 수 있다.In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification as long as it conforms to the enforcement laws at the time of manufacture and distribution. For example, it is a health functional food according to Korea's "Health Functional Food Act", or confectionery, beans, tea, and beverages according to each food type in the Food Ordinance of Korea "Food Sanitation Act" (Ministry of Food and Drug Safety Notification "Food Standards and Specifications") , special purpose food, and the like.
본 발명의 식품 조성물에는 그 유효성분 이외에 식품첨가물이 포함될 수 있다. 식품첨가물은 일반적으로 식품을 제조, 가공 또는 보존함에 있어 식품에 첨가되어 혼합되거나 침윤되는 물질로서 이해될 수 있는데, 식품과 함께 매일 그리고 장기간 섭취되므로 그 안전성이 보장되어야 한다. 식품의 제조·유통을 규율하는 각국 법률(한국에서는 「식품위생법」임)에 따른 식품첨가물공전에는 안전성이 보장된 식품첨가물이 성분 면에서 또는 기능 면에서 한정적으로 규정되어 있다. 한국 식품첨가물공전(식약처 고시 「식품첨가물 기준 및 규격」)에서는 식품첨가물이 성분 면에서 화학적 합성품, 천연 첨가물 및 혼합 제제류로 구분되어 규정되어 있는데, 이러한 식품첨가물은 기능 면에 있어서는 감미제, 풍미제, 보존제, 유화제, 산미료, 점증제 등으로 구분된다. The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives can be generally understood as substances that are added and mixed or infiltrated into food in manufacturing, processing, or preserving food. Food additives with guaranteed safety are limited in terms of ingredients or functions in the Food Additives Ordinance in accordance with the laws of each country that regulates the manufacture and distribution of food (“Food Sanitation Act” in Korea). In the Korean Food Additives Code (“Food Additive Standards and Specifications” announced by the Ministry of Food and Drug Safety), food additives are classified into chemically synthetic products, natural additives, and mixed preparations in terms of ingredients. It is divided into agents, preservatives, emulsifiers, acidulants, thickeners, etc.
감미제는 식품에 적당한 단맛을 부여하기 위하여 사용되는 것으로, 천연의 것이거나 합성된 것 모두 본 발명의 조성물에 사용할 수 있다. 바람직하게는 천연 감미제를 사용하는 경우인데, 천연 감미제로서는 옥수수 시럽 고형물, 꿀, 수크로오스, 프룩토오스, 락토오스, 말토오스 등의 당 감미제를 들 수 있다. The sweetener is used to impart appropriate sweetness to food, and both natural and synthetic ones may be used in the composition of the present invention. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.
풍미제는 맛이나 향을 좋게 하기 위하여 사용될 수 있는데, 천연의 것과 합성된 것 모두 사용될 수 있다. 바람직하게는 천연의 것을 사용하는 경우이다. 천연의 것을 사용할 경우에 풍미 이외에 영양 강화의 목적도 병행할 수 있다. 천연 풍미제로서는 사과, 레몬, 감귤, 포도, 딸기, 복숭아 등에서 얻어진 것이거나 녹차잎, 둥굴레, 댓잎, 계피, 국화 잎, 자스민 등에서 얻어진 것일 수 있다. 또 인삼(홍삼), 죽순, 알로에 베라, 은행 등에서 얻어진 것을 사용할 수 있다. 천연 풍미제는 액상의 농축액이나 고형상의 추출물일 수 있다. 경우에 따라서 합성 풍미제가 사용될 수 있는데, 합성 풍미제는 에스테르, 알콜, 알데하이드, 테르펜 등이 이용될 수 있다. Flavoring agents may be used to improve taste or aroma, and both natural and synthetic ones may be used. Preferably, it is a case where a natural thing is used. In the case of using a natural product, the purpose of nutritional enhancement in addition to flavor may be concurrently used. The natural flavoring agent may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or obtained from green tea leaves, honeysuckle, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo can be used. The natural flavoring agent may be a liquid concentrate or a solid extract. Optionally, a synthetic flavoring agent may be used, and the synthetic flavoring agent may be an ester, an alcohol, an aldehyde, a terpene, or the like.
보존제로서는 소듐 소르브산칼슘, 소르브산나트륨, 소르브산칼륨, 벤조산칼슘, 벤조산나트륨, 벤조산칼륨, EDTA(에틸렌디아민테트라아세트산) 등이 사용될 수 있고, 또 유화제로서는 아카시아검, 카르복시메틸셀룰로스, 잔탄검, 펙틴 등을 들 수 있으며, 산미료로서는 연산, 말산, 푸마르산, 아디프산, 인산, 글루콘산, 타르타르산, 아스코르브산, 아세트산, 인산 등이 사용될 수 있다. 산미료는 맛을 증진시키는 목적 이외에 미생물의 증식을 억제할 목적으로 식품 조성물이 적정 산도로 되도록 첨가될 수 있다.As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin, etc. are mentioned, and acidulant, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. can be used as an acidulant. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.
점증제로서는 현탁화 구현제, 침강제, 겔 형성제, 팽화제 등이 사용될 수 있다.As a thickening agent, a suspending agent, a settling agent, a gel former, a bulking agent, etc. can be used.
본 발명의 식품 조성물은 전술한 바의 식품첨가물 이외에, 기능성과 영양성을 보충, 보강할 목적으로 당업계에 공지되고 식품첨가물로서 안정성이 보장된 생리활성 물질이나 미네랄류를 포함할 수 있다.The food composition of the present invention may contain, in addition to the food additives described above, physiologically active substances or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutrition and guaranteed stability as food additives.
그러한 생리활성 물질로서는 녹차 등에 포함된 카테킨류, 비타민 B1, 비타민 C, 비타민 E, 비타민 B12 등의 비타민류, 토코페롤, 디벤조일티아민 등을 들 수 있으며, 미네랄류로서는 구연산 칼슘 등의 칼슘 제제, 스테아린산마그네슘 등의 마그네슘 제제, 구연산철 등의 철 제제, 염화 크롬, 요오드칼륨, 셀레늄, 게르마늄, 바나듐, 아연 등을 들 수 있다. Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. Minerals include calcium preparations such as calcium citrate, magnesium stearate Magnesium preparations, such as iron preparations, such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. are mentioned.
본 발명의 식품 조성물에는 전술한 바의 식품첨가물이 제품 유형에 따라 그 첨가 목적을 달성할 수 있는 적량으로 포함될 수 있다.In the food composition of the present invention, the food additives as described above may be included in an appropriate amount to achieve the purpose of the addition according to the product type.
본 발명의 식품 조성물에 포함될 수 있는 기타의 식품첨가물과 관련하여서는 각국 식품공전이나 식품첨가물 공전을 참조할 수 있다.In relation to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Ordinance of each country or the Food Additives Code.
본 발명의 조성물은 다른 구체적인 양태에 있어서는 약제학적 조성물로 파악될 수 있다.The composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.
본 발명의 약제학적 조성물은 유효성분 이외에 약제학적으로 허용되는 담체를 포함하여 당업계에 공지된 통상의 방법으로 투여 경로에 따라 경구용 제형 또는 비경구용 제형으로 제조될 수 있다. 여기서 투여 경로는 국소 경로, 경구 경로, 정맥 내 경로, 근육 내 경로, 및 점막 조직을 통한 직접 흡수를 포함하는 임의의 적절한 경로일 수 있으며, 두 가지 이상의 경로를 조합하여 사용할 수도 있다. 두 가지 이상 경로의 조합의 예는 투여 경로에 따른 두 가지 이상의 제형의 약물이 조합된 경우로서 예컨대 1차로 어느 한 약물은 정맥 내 경로로 투여하고 2차로 다른 약물은 국소 경로로 투여하는 경우이다. The pharmaceutical composition of the present invention may be prepared as an oral dosage form or a parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of the combination of two or more routes is a case in which two or more formulations of drugs according to the route of administration are combined. For example, one drug is first administered by an intravenous route and the other drug is secondarily administered by a local route.
약학적으로 허용되는 담체는 투여 경로나 제형에 따라 당업계에 주지되어 있으며, 구체적으로는 "대한민국약전"을 포함한 각국의 약전을 참조할 수 있다. Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically, reference may be made to the pharmacopoeia of each country including the "Korea Pharmacopoeia".
본 발명의 약제학적 조성물이 경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 분말, 과립, 정제, 환제, 당의정제, 캡슐제, 액제, 겔제, 시럽제, 현탁액, 웨이퍼 등의 제형으로 제조될 수 있다. 이때 적합한 담체의 예로서는 락토스, 글루코스, 슈크로스, 덱스트로스, 솔비톨, 만니톨, 자일리톨 등의 당류, 옥수수 전분, 감자 전분, 밀 전분 등의 전분류, 셀룰로오스, 메틸셀룰로오스, 에틸셀룰로오스, 나트륨 카르복시메틸셀룰로오스, 하이드록시프로필메틸셀룰로오스 등의 셀룰로오스류, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 마그네슘 스테아레이트, 광물유, 맥아, 젤라틴, 탈크, 폴리올, 식물성유, 에탄올, 글리세롤 등을 들 수 있다. 제제화할 경우 필요에 따라적절한 결합제, 윤활제, 붕해제, 착색제, 희석제 등을 포함시킬 수 있다. 적절한 결합제로서는 전분, 마그네슘 알루미늄 실리케이트, 전분 페이스트, 젤라틴, 메틸셀룰로스, 소듐 카복시메틸셀룰로스, 폴리비닐피롤리돈, 글루코스, 옥수수 감미제, 소듐 알지네이트, 폴리에틸렌 글리콜, 왁스 등을 들 수 있고, 윤활제로서는 올레산나트륨, 스테아르산나트륨, 스테아르산마그네슘, 벤조산나트륨, 초산나트륨, 염화나트륨, 실리카, 탈쿰, 스테아르산, 그것의 마그네슘염과 칼슘염, 폴리데틸렌글리콜 등을 들 수 있으며, 붕해제로서는 전분, 메틸 셀룰로스, 아가(agar), 벤토나이트, 잔탄검, 전분, 알긴산 또는 그것의 소듐 염 등을 들 수 있다. 또 희석제로서는 락토즈, 덱스트로즈, 수크로즈, 만니톨, 소비톨, 셀룰로스, 글라이신 등을 들 수 있다. When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art together with a suitable carrier It can be prepared in a formulation such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Cellulose such as hydroxypropylmethylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, glycerol and the like. In the case of formulation, an appropriate binder, lubricant, disintegrant, colorant, diluent, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, and the like, and lubricants include sodium oleate. , sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium salt and calcium salt, polyethylene glycol, etc., and the disintegrant includes starch, methyl cellulose, agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, and the like.
본 발명의 약제학적 조성물이 비경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 주사제, 경피 투여제, 비강 흡입제 및 좌제의 형태로 제제화될 수 있다. 주사제로 제제화할 경우 적합한 담체로서는 수성 등장 용액 또는 현탁액을 사용할 수 있으며, 구체적으로는 트리에탄올 아민이 함유된 PBS(phosphate buffered saline)나 주사용 멸균수, 5% 덱스트로스 같은 등장 용액 등을 사용할 수 있다. 경피 투여제로 제제화할 경우 연고제, 크림제, 로션제, 겔제, 외용액제, 파스타제, 리니멘트제, 에어롤제 등의 형태로 제제화할 수 있다. 비강 흡입제의 경우 디클로로플루오로메탄, 트리클로로플루오로메탄, 디클로로테트라플루오로에탄, 이산화탄소 등의 적합한 추진제를 사용하여 에어로졸 스프레이 형태로 제제화할 수 있으며, 좌제로 제제화할 경우 그 담체로는 위텝솔(witepsol), 트윈(tween) 61, 폴리에틸렌글리콜류, 카카오지, 라우린지, 폴리옥시에틸렌 소르비탄 지방산 에스테르류, 폴리옥시에틸렌 스테아레이트류, 소르비탄 지방산 에스테르류 등을 사용할 수 있다.When the pharmaceutical composition of the present invention is prepared for parenteral use, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories together with suitable carriers according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or isotonic solution such as 5% dextrose may be used. . When formulated for transdermal administration, it can be formulated in the form of an ointment, a cream, a lotion, a gel, an external solution, a pasta agent, a liniment agent, an air roll, and the like. In the case of nasal inhalants, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, and the like. witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like can be used.
약제학적 조성물의 구체적인 제제화와 관련하여서는 당업계에 공지되어 있으며, 예컨대 문헌[Remington's Pharmaceutical Sciences(19th ed., 1995)] 등을 참조할 수 있다. 상기 문헌은 본 명세서의 일부로서 간주 된다.Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). This document is considered a part of this specification.
본 발명의 약제학적 조성물의 바람직한 투여량은 환자의 상태, 체중, 성별, 연령, 환자의 중증도, 투여 경로에 따라 1일 0.001mg/kg ~ 10g/kg 범위, 바람직하게는 0.001mg/kg ~ 1g/kg 범위일 수 있다. 투여는 1일 1회 또는 수회로 나누어 이루어질 수 있다. 이러한 투여량은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 해석되어서는 아니 된다. A preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, severity of the patient, and the route of administration. It can be in the range /kg. Administration may be performed once or divided into several times a day. These dosages should not be construed as limiting the scope of the invention in any respect.
본 발명의 조성물은 또 다른 구체적인 양태에 있어서, 화장료 조성물로 파악할 수 있다. In another specific embodiment, the composition of the present invention can be identified as a cosmetic composition.
본 발명의 조성물이 화장료 조성물로 파악될 경우에도 그 화장료 조성물은 그 용도상, 법률상 임의의 제품 구분을 띨 수 있으며, 구체적으로 피부 트러블 개선, 아토피 피부염 개선 등의 용도를 가진 기능성 화장품, 비기능성 일반 화장품 등일 수 있다. 제품 형태에 있어서도 임의의 제품 형태를 띨 수 있는데, 구체적으로 용액, 현탁액, 유탁액, 페이스트, 젤, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 스프레이 등의 제품 형태를 띨 수 있다. 구체적인 제품 형태에 있어서는 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 폼, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형 등일 수 있다.Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be classified into any product in terms of its use or by law, and specifically functional cosmetics, non-functional products having uses such as improvement of skin troubles and improvement of atopic dermatitis It may be general cosmetics and the like. In terms of product form, it may take any product form, and specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can take the form of products such as foundation and spray. In the specific product form, it may be in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.
본 발명의 화장료 조성물은 그 유효성분 이외에 화장료 조성물에 통상적으로 이용되는 성분들, 예컨대, 안정화제, 용해화제, 계면활성제, 비타민, 색소 및 향료와 같은 통상적인 보조제, 및 담체를 포함할 수 있다. The cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the active ingredient, for example, conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and fragrances, and carriers.
본 발명의 제형이 페이스트, 크림 또는 젤인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되는데, 구체적으로 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜, 소르비탄의 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, etc. may be used.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르, 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 등이 이용될 수 있다.When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar, etc. can be used.
본 발명의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester and the like may be used.
본 발명의 화장료 조성물은 항산화, 항균 활성을 나타내는 그 유효성분을 포함하는 것을 제외하고는 당업계에 통상적으로 행하여지는 화장료 조성물의 제조방법에 따라 제조할 수 있다.The cosmetic composition of the present invention can be prepared according to the method for preparing a cosmetic composition conventionally performed in the art, except that it contains the active ingredient exhibiting antioxidant and antibacterial activity.
전술한 바와 같이, 본 발명에 따르면, 여뀌바늘 추출물을 이용한 항산화 또는 항균용 조성물을 제공할 수 있다.As described above, according to the present invention, it is possible to provide a composition for anti-oxidation or antibacterial using the extract of chrysanthemum.
본 발명의 조성물은 식품, 약품 또는 화장품으로 제품화될 수 있다.The composition of the present invention may be commercialized as food, drug or cosmetic.
도 1 및 도 2는 여뀌바늘 추출물의 항산화 활성에 대한 실험 결과이다.
도 3 내지 도4는 여뀌바늘 추출물의 항균 활성에 대한 실험 결과이다.1 and 2 are experimental results for the antioxidant activity of the extract of yew needles.
3 to 4 are experimental results for the antibacterial activity of the extract of yew needles.
이하 본 발명을 실시예 및 실험예를 참조하여 설명한다. 그러나 본 발명의 범위가 이러한 실시예 및 실험예에 한정되는 것은 아니다.Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.
<실시예> 여뀌바늘 잎 추출물과 줄기 추출물의 제조<Example> Preparation of leaf extract and stem extract
여뀌바늘 잎 건조분말에 30배 중량의 증류수를 가하고 (120℃에서 2시간) 열수추출한 후 여과하고 추출용매를 제거하여 동결건조를 거쳐 여뀌바늘 잎 추출물을 제조하였다. After adding 30 times the weight of distilled water (at 120° C. for 2 hours) to the dry powder of dandelion leaves, hot water extraction was performed, followed by filtration, removal of the extraction solvent, and freeze-drying to prepare a quince needle leaf extract.
여뀌바늘 줄기 추출물도 상기와 동일하게 열수추출하여 제조하였다.The extract of the genus needles stems was prepared by hot water extraction in the same manner as above.
<실험예> 항산화 활성 및 항균 활성<Experimental Example> Antioxidant activity and antibacterial activity
<실험예 1> 항산화 활성 실험<Experimental Example 1> Antioxidant activity experiment
<실험예 1-1> DPPH assay에 의한 항산화 활성 실험 <Experimental Example 1-1> Antioxidant activity experiment by DPPH assay
DPPH(1,1-diphenyl-2-picryl hydrazyl) 법에 따른 여뀌바늘 추출물의 항산화 활성은 다음과 같이 확인하였다.The antioxidative activity of the extracts of chrysanthemums according to the DPPH (1,1-diphenyl-2-picryl hydrazyl) method was confirmed as follows.
DPPH 라디칼 소거활성은 DPPH가 에탄올상에서 발생시키는 자유 라디칼에 대한 시료의 소거능을 측정하는 방법으로 메탄올에 녹인 0.2 mM DPPH 용액 0.1 ml에 일정 농도로 희석된 각 추출물 0.1 ml을 넣고 10초간 혼합하였다. 그리고 빛을 차단하여 30분간 상온에서 반응시킨 뒤 분광광도계를 이용하여 517 nm의 파장에서 흡광도를 측정하였으며, 시료 무처리구 대비 백분율로 결과를 산출하여 도 1에 나타내었다. 도 1을 참조하여 보면 여뀌바늘 잎 추출물과 줄기 추출물 모두 농도 의존적으로 DPPH 소거 활성을 보임을 알 수 있다.DPPH radical scavenging activity is a method of measuring the scavenging ability of a sample for free radicals generated by DPPH in ethanol. 0.1 ml of each extract diluted to a certain concentration in 0.1 ml of a 0.2 mM DPPH solution dissolved in methanol was added and mixed for 10 seconds. Then, after blocking the light and reacting at room temperature for 30 minutes, the absorbance was measured at a wavelength of 517 nm using a spectrophotometer. Referring to FIG. 1 , it can be seen that both the leaf extract and the stem extract show DPPH scavenging activity in a concentration-dependent manner.
<실험예 1-2> 환원력 측정 실험<Experimental Example 1-2> reducing power measurement experiment
환원력 측정은 potassium ferricyanide법을 이용하여 측정하였다. 각 추출물 1.0 ml (1 mg/ml)에 200 mM phosphate buffer(pH 6.6)와 1% potassium ferricyanide 용액을 각각 2.5ml씩 차례로 첨가하여 교반한 후 50℃에서 20분간 반응시켰다. 반응액은 10% trichloroacetic acid 2.5 ml을 가하여 3,500 rpm에서 10분간 원심분리한 후 상등액 2.5 ml에 증류수 2.5 ml과 ferric chloride 용액 0.5 ml을 첨가하여 혼합한 후 Multimode microplate reader (Varioskan LUX, ThermoFisher Scientific, Inc. Co. MA, USA)를 이용하여 700 nm에서 흡광도를 측정하였다. 분석한 결과는 absorbance of 700 nm로 하여 도 2에 나타내었다. 도 2를 참조하여 보면, 도 1의 결과와 마찬가지로 여뀌바늘 잎 추출물과 줄기 추출물 모두 농도 의존적으로 환원력이 증가함을 알 수 있다.The reducing power was measured using the potassium ferricyanide method. To 1.0 ml (1 mg/ml) of each extract, 2.5 ml of 200 mM phosphate buffer (pH 6.6) and 1% potassium ferricyanide solution were sequentially added and stirred, followed by reaction at 50° C. for 20 minutes. To the reaction solution, 2.5 ml of 10% trichloroacetic acid was added, centrifuged at 3,500 rpm for 10 minutes, and 2.5 ml of distilled water and 0.5 ml of ferric chloride solution were added to 2.5 ml of the supernatant and mixed, followed by a multimode microplate reader (Varioskan LUX, ThermoFisher Scientific, Inc.). (Co. MA, USA) was used to measure the absorbance at 700 nm. The analysis result was shown in FIG. 2 as an absorbance of 700 nm. Referring to FIG. 2 , it can be seen that the reducing power is increased in a concentration-dependent manner for both the genus oleifera leaf extract and the stem extract, as in the result of FIG. 1 .
<실험예 2> 항균 활성 실험<Experimental Example 2> Antibacterial activity test
<실험예 2-1> 디스크 확산법(Disc diffusion assay)에 따른 항균 활성 실험<Experimental Example 2-1> Antibacterial activity test according to the disc diffusion assay
항균 효과를 확인하기 위하여 사용한 균주는 아래 표 1과 같이 그람양성균인 스타필로코쿠스 아우레우스(Staphylococcus aureus ATCC 29213), 메티실린 내성 황색포도상구균(MRSA) 2종과 그람음성균인 대장균(Escherichia coli) 1종을 사용하였으며, 배지는 LB broth 및 LB agar를 사용하여 37℃에서 조건에서 배양하였다. 그람음성균인 대장균은 가천대학교에서 분양받아 사용하였고, 그람양성균인 스타필로코쿠스 아우레우스는 한국미생물보존센터에서 분양받은 균주를 사용하였으며, MRSA는 가천대학교 길병원에서 분양 받아 사용하였다. The strains used to confirm the antibacterial effect are Gram-positive bacteria, Staphylococcus aureus ATCC 29213, as shown in Table 1 below; Two methicillin-resistant Staphylococcus aureus (MRSA) and one gram-negative Escherichia coli were used, and the medium was cultured at 37°C using LB broth and LB agar. Escherichia coli, a gram-negative bacterium, was purchased from Gachon University, Staphylococcus aureus, a gram-positive bacterium, a strain acquired from the Korea Microbial Conservation Center was used, and MRSA was purchased and used from Gachon University Gil Hospital.
여뀌바늘 추출물의 항균 활성은 paper disc를 이용한 디스크 확산법(Disc diffusion assay)으로 확인하였다. 균주 접종을 위해 고온 고압 멸균기를 통해 멸균된 배양배지 3 ㎖에 배양균을 이식하고 37 ℃ 에서 24 시간 동안 진탕 배양 하였다. 배양된 균주를 1 x 105 cfu/㎖ 로 희석하여 agar plate에 고루 퍼지도록 도말하였다. 멸균된 paper disc에 추출물을 농도별로 접종하여 배지 표면에 가볍게 놓은 후 37 ℃ 에서 24 시간 동안 배양하면서 생육저해환 (clear zone) 생성 유무를 확인하였다. The antibacterial activity of the extracts of chrysanthemum needles was confirmed by a disc diffusion assay using a paper disc. For strain inoculation, the culture medium was transplanted into 3 ml of sterilized culture medium through a high-temperature high-pressure sterilizer, and cultured with shaking at 37° C. for 24 hours. The cultured strain was diluted to 1 x 10 5 cfu/ml and spread evenly on an agar plate. After inoculating the extracts by concentration on sterilized paper discs, lightly placing them on the surface of the medium, and culturing at 37°C for 24 hours, it was checked whether or not a clear zone was generated.
결과를 도 3(잎) 및 도 4(줄기)에 여뀌바늘 잎 추출물과 줄기 추출물 각각의 생육 저해환(clear zone) 사진을 나타내는 한편 아래의 표 2(잎) 및 표 3(줄기)에 생육 저해환의 크기를 측정하여 나타내었다.The results are shown in Figures 3 (leaf) and 4 (stem) of the growth inhibition ring (clear zone) of each of the leaf extract and stem extract, while the growth inhibition is shown in Table 2 (leaf) and Table 3 (stem) below. The size of the ring was measured and shown.
여뀌바늘 잎 추출물과 줄기 추출물 모두 3가지 미생물에 대해 항균 활성을 나타내었고, 특히 잎 추출물의 항균 활성이 우수하였다. 1,2-Hexanediol은 양성대조군으로 사용되었다.Both the leaf extract and the stem extract showed antibacterial activity against three microorganisms, and the leaf extract was particularly excellent in antibacterial activity. 1,2-Hexanediol was used as a positive control.
<실험예 2-2> 최소저해농도(MIC) 측정 실험<Experimental Example 2-2> Minimum inhibitory concentration (MIC) measurement experiment
실험 균주에 대한 추출물의 최소성장억제농도 (MIC) 측정을 위해 추출물을 멸균된 LB broth 로 1/2 씩 최소 농도가 0.004%가 될 때까지 희석하고 96 well plate 에 20 ㎕ 씩 넣은 후 각각의 균주를 1 x 105 cfu/㎖ 180 ㎕ 를 첨가하였다. 37 ℃ 항온기에서 24 시간 동안 배양한 후 흡광광도계를 통해 595 nm 파장에서 흡광도를 측정하여 접종 균주가 성장하지 않은 추출물 농도를 MIC로 하였다. To measure the minimum growth inhibitory concentration (MIC) of the extract for the test strain, dilute the extract by 1/2 with sterile LB broth until the minimum concentration is 0.004%, and then put 20 μl into a 96 well plate, and then each strain 1 x 10 5 cfu/ml 180 μl was added. After culturing for 24 hours in a 37 °C incubator, absorbance was measured at a wavelength of 595 nm through an absorbance meter, and the concentration of the extract in which the inoculated strain was not grown was used as the MIC.
결과를 아래의 표 4에 나타내었다. 여뀌바늘 잎 추출물의 최소 저해 농도는 1mg/mL, 여뀌바늘 줄기 추출물의 최소 저해 농도는 2mg/mL로 나타났다. The results are shown in Table 4 below. The minimum inhibitory concentration of the genus leaf extract was 1 mg/mL, and the minimum inhibitory concentration of the genus oleifera stem extract was 2 mg/mL.
Claims (7)
A composition for antioxidants comprising the extract of chrysanthemum needles as an active ingredient.
상기 여뀌바늘 추출물은 여뀌바늘의 잎 또는 줄기의 물, 에탄올 또는 이들의 혼합용매 추출물인 것을 특징으로 하는 조성물.
The method of claim 1,
The extract is a composition characterized in that the extract of water, ethanol, or a mixed solvent thereof of the leaves or stems of the cypress needles.
An antibacterial composition comprising an extract of chrysanthemum needles as an active ingredient.
상기 항균은 황색포도상구균(Staphylococcus aureus), 메티실린 내성 황색포도상구균(Methicillin-resistant Staphylococcus aureus, MRSA), 및 대장균에 대해 항균 활성인 것을 특징으로 하는 조성물.
4. The method of claim 3,
The antibacterial is Staphylococcus aureus ( Staphylococcus aureus ), methicillin-resistant Staphylococcus aureus (Methicillin-resistant Staphylococcus aureus , MRSA), and the composition characterized in that the antibacterial activity against E. coli.
상기 여뀌바늘 추출물은 여뀌바늘의 잎 또는 줄기의 물, 에탄올 또는 이들의 혼합용매 추출물인 것을 특징으로 하는 조성물.
4. The method of claim 3,
The extract is a composition characterized in that the extract of water, ethanol, or a mixed solvent thereof of the leaves or stems of the cypress needles.
상기 조성물은 식품 조성물인 것을 특징으로 하는 조성물.
4. The method of claim 3,
The composition is a food composition, characterized in that the composition.
상기 조성물은 화장품 조성물인 것을 특징으로 하는 조성물.
4. The method of claim 3,
The composition is a cosmetic composition, characterized in that the composition.
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