KR101411998B1 - Preparation of opuntia humifusa extract using enzymes under ultra high pressure - Google Patents
Preparation of opuntia humifusa extract using enzymes under ultra high pressure Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/06—Enzymes
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A23V2200/00—Function of food ingredients
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2300/00—Processes
- A23V2300/28—Hydrolysis, degree of hydrolysis
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Abstract
본 발명은 초고압 조건 하 천년초에 효소를 처리하여 가수분해하는 단계를 포함하는 천년초 추출물의 제조 방법에 대한 것이다. 본 발명의 제조 방법으로 제조된 천년초 추출물은 천년초의 점성이 실질적으로 제거되어 이용이 용이할 뿐 아니라, 항산화 활성, 라디칼 소거능이 우수한 이점이 있다.The present invention relates to a method for producing a quillaine starch extract comprising hydrolyzing an enzyme by treatment with an enzyme at a high-pressure condition. The tsunami extract prepared by the method of the present invention is advantageous in that it is easy to use because the viscosity of the tsunami seeds is substantially eliminated, and the antioxidant activity and the radical scavenging ability are excellent.
Description
본 발명은 천년초의 점성을 제거하여 항산화 활성이 높은 천년초 추출물을 제거하는 방법에 대한 것이다.The present invention relates to a method for eliminating milky water extracts having a high antioxidative activity by eliminating viscous water of milky water.
천년초(Opuntia humifusa)는 충남 등지에서 자생하는 한국 토종 선인장이다. 흔히들 천년초는 국문 명칭이 비슷한 백년초(Opuntia ficus - indica var saboten)와 혼동되는 경우가 많으나 양자는 그 특성이 상이하다. 즉, 일반적으로 손바닥 선인장으로 널리 알려진 백년초가 1-2 m 크기로 자라며 길고 굵은 가시가 많은 반면 천년초는 솜털 가시를 갖고 약 30 cm 크기로 자란다. 제주도에서 재배되는 백년초는 열대지방이 원산지로 저온에 약하여 노지재배가 어려운 반면, 한국 토종 선인장인 천년초는 휴면기인 겨울에도 노지에서 생존하며 병충해에 강한 특징이 있다.
Opuntia humifusa is a Korean native cactus native to Chungcheongnam-do. Heunhideul cheonnyeoncho is the Korean name similar baeknyeoncho (Opuntia ficus - indica there saboten ) are often confused with the characteristics of the two are different. In other words, commonly known as the palm cactus, the perennial plant grows 1-2 m in length, while the long and thick thorns grow, whereas the millennium plant grows to about 30 cm in size with fluffy spiny. The cultivated white perennial plant in Jeju Island is weak in low temperature due to its tropical origin. On the other hand, Korean native cactus, Chunnungcho, survives in the winter, which is a dormant period, and is resistant to pests and diseases.
귀화식물인 백년초에 비하여 천년초의 활성에 대하여는 많은 연구가 이루어지지 않은 편이며, 식품, 화장품 등으로 가공, 상용화 역시 거의 이루어지지 않고 있다. 천년초의 가공 및 이용이 활발하지 못한 주된 이유는 천년초의 주성분인 펙틴에 의한 점성 때문으로, 천년초를 이용한 다양한 제품을 개발하기 위하여는 천년초의 소재화를 위한 추출방법이 개선될 것이 우선적으로 요구된다.
There is not much research on the activity of the millennium early in comparison with the naturalized plant, which is a natural plant, and the processing and commercialization of it by food, cosmetics and the like are hardly achieved. The main reason that the processing and use of the millenniace is not active is due to the viscosity due to pectin which is the main ingredient of millennia. Therefore, in order to develop various products using millennium, it is firstly demanded that the extraction method for the millennia.
현재까지 천년초의 점성 제거를 위한 시도로는 여과, 효소 가수분해, 발효, 천년초 동결 후 분말화 방법 등이 제시되어 왔는데(대한민국 등록 특허 제 10-0763797호), 여과나 분말화 등의 방법은 점성이 효과적으로 제거되지 않고, 일반적인 효소 가수분해의 경우 점성이 충분히 제거되기까지 시간이 많이 소요되거나, 다량의 효소가 필요하여 비용이 증가하는 문제점이 있었다.To date, efforts to remove viscous water of the millennia have been proposed (filtration, enzymatic hydrolysis, fermentation, and post-freeze-thawing methods) (Korean Patent No. 10-0763797) The enzymatic hydrolysis does not effectively remove the enzyme. In the case of general enzyme hydrolysis, it takes a long time until the viscosity is sufficiently removed or a large amount of enzyme is required, which increases the cost.
본 발명의 목적은 천년초의 점성을 빠른 시간 내에 효과적으로 제거하여 라디칼 소거능이 증진된 천년초 추출물을 제조하는 방법을 제공하는 것이다.It is an object of the present invention to provide a method for manufacturing a quinceanil seed extract having improved radical scavenging ability by effectively removing the viscosity of the millennia early.
상기 목적을 달성하기 위하여 본 발명은 초고압 조건 하 천년초에 효소를 처리하여 가수분해하는 단계를 포함하는 천년초 추출물의 제조 방법을 제공한다.In order to accomplish the above object, the present invention provides a method for preparing a tsunami extract comprising the step of hydrolyzing an enzyme at an early stage under an ultrahigh pressure condition.
본 발명의 천년초 추출물의 제조 방법은 천년초의 점성을 빠른 시간 내에 충분히 제거하면서도 항산화 효능 및 라디칼 소거능이 우수한 천년초 추출물을 제조할 수 있다.The method of the present invention for producing the mushroom extract of the present invention can produce a mushroom extract having excellent antioxidative activity and radical scavenging ability while rapidly removing the viscosity of the mushroom starch.
도 1은 가수분해 효소 및 압력에 따른 천년초 추출물의 라디칼 소거능을 나타낸다.
도 2는 가수분해 효소 및 압력에 따른 천년초 추출물의 점도를 나타낸다.
도 3은 펙티넥스 및 사이토레이즈 PCL 5의 비율에 따른 효소 혼합물의 천년초 점성 제거능을 나타낸다.
도 4는 초고압 및 혼합 효소 처리 여부에 따른 천년초 추출물의 고형분 및 폴리페놀 함량 차이를 나타낸다.
도 5는 초고압 및 혼합 효소 처리 여부에 따른 천년초 추출물의 총 당 및 산성당 함량 차이를 나타낸다.
도 6은 초고압 및 혼합 효소 처리 여부에 따른 천년초 추출물의 라디칼 소거능 차이를 나타낸다.
도 7은 초고압 및 혼합 효소 처리 여부에 따른 천년초 추출물의 항보체 활성 차이를 나타낸다.
도 8은 초고압 및 혼합 효소 처리 여부에 따른 천년초 추출물의 점도 차이를 나타낸다.
도 9는 초고압 조건 하 혼합 효소 처리 시간에 따른 천년초 추출물의 점도 변화(a), 총 당 함량의 변화(b) 및 산성당 함량의 변화(c)를 나타낸다.
도 10은 초고압 조건 하 혼합 효소 처리 시간에 따른 천년초 추출물의 DPPH 및 ABTS 라디칼 소거능의 변화를 나타낸다.Fig. 1 shows the radical scavenging ability of the acarola extract according to hydrolytic enzymes and pressure.
Fig. 2 shows the viscosity of the acarola extract according to hydrolytic enzymes and pressure.
Figure 3 shows the ability of the enzyme mixture to remove viscous water in the millennia depending on the ratio of pectinex and
FIG. 4 shows the difference in solid content and polyphenol content of the quinternial root extract according to the treatment with ultrahigh pressure and mixed enzyme.
FIG. 5 shows the differences in the total sugar and acid content of the Korean acantholyticus extract according to the treatment with ultrahigh pressure and mixed enzyme.
FIG. 6 shows the difference in radical scavenging ability of the quinternial root extract according to the treatment with ultrahigh pressure and mixed enzyme.
FIG. 7 shows the difference in antifouling activity between the extracts of Cheolwon-gil on the basis of ultrahigh-pressure and mixed enzyme treatment.
FIG. 8 shows the viscosity difference of the Cheonnyuck seed extract according to whether ultra-high pressure or mixed enzyme treatment was performed.
9 shows changes in viscosity (a), change in total sugar content (b) and change in acid content (c) of the Cheonjiyang extract according to the time of the mixed enzyme treatment under the high pressure condition.
FIG. 10 shows changes in DPPH and ABTS radical scavenging activity of the quinternial root extract according to time of mixed enzyme treatment under ultrahigh pressure.
본 발명은 초고압 조건 하 천년초에 효소를 처리하여 가수분해하는 단계를 포함하는 천년초 추출물의 제조 방법을 제공한다.The present invention provides a method for preparing a tsunami extract comprising the step of hydrolyzing an enzyme at an early stage under an ultrahigh pressure condition.
또한 본 발명은 상기 방법으로 제조된 천년초 추출물을 제공한다.In addition, the present invention provides a tsunami extract prepared by the above method.
또한 본 발명은 상기 천년초 추출물을 유효성분으로 포함하는 식품 조성물 및 화장료 조성물을 제공한다.
In addition, the present invention provides a food composition and a cosmetic composition comprising the citrus extract as an active ingredient.
본 발명의 이점 및 특징, 그리고 그것들을 달성하는 방법은 상세하게 후술되어 있는 실험예들을 참조하면 명확해질 것이다. 그러나, 본 발명은 이하에서 개시되는 실험예들에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 것이며, 단지 본 실험예들은 본 발명의 개시가 완전하도록 하며, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이며, 본 발명은 청구항의 범주에 의해 정의될 뿐이다.
Advantages and features of the present invention, and methods of achieving them, will become apparent with reference to the following detailed examples. However, it should be understood that the present invention is not limited to the following examples, but may be embodied in many different forms and should not be construed as limited to the exemplary embodiments set forth herein. To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.
천년초Millennium
본 발명의 천년초는 오푼티아 후미푸사(Opuntia humifusa)이다. 본 발명의 천년초는 수세 및 가시를 제거하고 물을 첨가하여 마쇄한 상태로 가수분해에 사용하는 것이 바람직하나, 이에 제한되는 것은 아니다.
The milky candle of the present invention is Opuntia humifusa . In the present invention, it is preferable to use water for hydrolysis in the milled state by removing water and visibility and adding water thereto, but it is not limited thereto.
초고압Ultrahigh pressure
본 발명의 초고압은 100 내지 1,000 MPa이 될 수 있으며, 바람직하게는 100 내지 300 MPa이고, 더욱 바람직하게는 100 내지 200 MPa이나, 이에 제한되는 것은 아니다.
The ultrahigh pressure of the present invention may be 100 to 1,000 MPa, preferably 100 to 300 MPa, and more preferably 100 to 200 MPa.
효소enzyme
본 발명의 효소는 가수분해효소이다. 예컨대, 본 발명의 효소는 셀룰레이즈, 펙티네이즈, 헤미셀룰레이즈, 아라비네이즈, 베타-글루카네이즈, 자일라네이즈, 알파-아밀레이즈, 글루코아밀레이즈, 폴리갈락투로네이즈로 구성된 군으로부터 선택되는 하나 이상일 수 있다. 또한 본 발명의 효소는 에코니테이즈 CE, 라피데이즈, 비스코자임, 울트라플로 L, 사이토레이즈 PCL 5, 옵티덱스 L-400, 셀루클라스트 1.5L 및 펙티넥스 5XL로 구성되는 군으로부터 선택되는 하나 이상일 수 있다.The enzyme of the present invention is a hydrolytic enzyme. For example, the enzyme of the present invention is selected from the group consisting of cellulase, pectinase, hemicellulase, arabinase, beta-glucanase, xylenes, alpha-amylase, glucoamylase, polygalacturonose It can be more than one. In addition, the enzyme of the present invention may be at least one enzyme selected from the group consisting of Econidase CE, Rapidays, Viscomzyme, Ultraflow L, CytoRaze PCL 5, Optidex L-400, Celluclast 1.5 L and Pectinex 5XL .
바람직하게는 본 발명의 효소는 펙티넥스 및 사이토레이즈 PCL 5으로 구성되는 군으로부터 선택되는 하나 이상이며, 더욱 바람직하게는 본 발명의 효소는 펙티넥스 및 사이토레이즈 PCL 5의 혼합 효소이다. 더더욱 바람직하게는 본 발명의 효소는 펙티넥스 및 사이토레이즈를 3:1 내지 1:3의 중량비로 혼합한 혼합 효소이고, 가장 바람직하게는 펙티넥스 및 사이토레이즈 PCL 5를 동량으로 혼합한 혼합 효소이나 이에 제한되는 것은 아니다.
Preferably, the enzyme of the present invention is at least one selected from the group consisting of pectinex and
천년초Millennium 추출물 extract
본 발명의 천년초 추출물은 천년초의 점성이 실질적으로 제거된 것이다. 상기 "실질적으로 제거"란 천년초 추출물을 원료로 하여 일반적인 공정에 제공할 수 있는 상태를 가리킨다. 예컨대, 본 발명의 천년초 추출물은 15 cP 이하의 점도를 가지며, 더욱 바람직하게는 10 cP 이하의 점도를 갖고, 더더욱 바람직하게는 5 cP 이하의 점도를 갖는다.The tsunami extract of the present invention is substantially removed from the tsunami-annual viscosity. The term "substantially eliminated" refers to a state in which the extract is available as a raw material and can be supplied to a general process. For example, the quinterniaceae extract of the present invention has a viscosity of 15 cP or less, more preferably 10 cP or less, and still more preferably 5 cP or less.
또한 본 발명의 천년초 추출물은 항산화 활성 또는 자유 라디칼 소거능을 갖는다.
The present invention also provides an antioxidant activity or free radical scavenging ability.
식품 조성물Food composition
본 발명의 식품 조성물은 건강보조식품, 건강기능식품, 기능성 식품 등이나 이에 제한되는 것은 아니며, 천연식품, 가공식품, 일반적인 식자재 등에 본 발명의 천년초 추출물을 첨가하는 것도 포함된다. 또한 본 발명의 식품 조성물은 동물용 사료를 포함한다.
The food composition of the present invention is not limited to a health supplement food, a health functional food, a functional food and the like, but it also includes the addition of the quintern herb extract of the present invention to natural foods, processed foods, and general food ingredients. The food composition of the present invention also includes animal feed.
본 발명의 식품 조성물은, 상기 천년초 추출물을 그대로 첨가하거나 다른 식품 또는 식품 조성물과 함께 사용될 수 있으며, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그의 사용 목적에 따라 적절하게 결정될 수 있다. 일반적으로, 본 발명의 천년초 추출물은, 식품 또는 음료의 제조 시에 식품 또는 음료의 원료 100 중량부에 대하여 0.1 내지 90 중량부, 바람직하게는 1 내지 60 중량부 첨가될 수 있다.
The food composition of the present invention can be used as it is, or can be used in combination with other food or food compositions, and can be suitably used according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use thereof. Generally, the quintern herb extract of the present invention may be added in an amount of 0.1 to 90 parts by weight, preferably 1 to 60 parts by weight, based on 100 parts by weight of the raw material of the food or beverage when the food or beverage is prepared.
상기 식품의 종류에는 특별한 제한은 없다. 상기 천년초 추출물을 포함하는 식품 조성물은 정제, 경질 또는 연질 캅셀제, 액제, 현탁제 등과 같은 경구투여용 제제의 형태로 이용될 수 있으며, 이들 제제는 허용 가능한 통상의 담체, 예를 들어 경구투여용 제제의 경우에는 부형제, 결합제, 붕해제, 활택제, 가용화제, 현탁화제, 보존제 또는 증량제 등을 사용하여 조제할 수 있다. There is no particular limitation on the kind of the food. The food composition comprising the snakebird extract may be used in the form of tablets, hard or soft capsules, liquids, suspensions, and the like, which may contain acceptable conventional carriers, for example, oral preparations A binder, a disintegrant, a lubricant, a solubilizing agent, a suspending agent, a preservative, or an extender, for example.
상기 천년초 추출물을 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제, 기타 영양제 등을 들 수 있으나 이들 종류의 식품으로 제한되는 것은 아니다.
Examples of foods that can be supplemented with the citrus jujube extract include dairy products such as meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, drinks, tea, , Alcoholic beverages and vitamin complexes, and other nutrients, but are not limited to these kinds of foods.
화장료Cosmetics 조성물 Composition
본 발명의 화장료 조성물은, 당업계에서 통상적으로 제조되는 임의의 제형으로 제조될 수 있으며 용액, 현탁액, 유탁액, 페이스트, 젤, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 제한되는 것은 아니다. 또한 본 발명의 화장료 조성물은 유연화장수, 영양화장수, 영양크림, 마사지크림, 에센스, 아이크림, 클렌징크림, 클렌징폼, 클렌징워터, 팩, 스프레이 또는 파우더 제형으로 제조될 수도 있다.
The cosmetic composition of the present invention may be prepared in any formulations conventionally produced in the art and may be formulated into solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant- A foundation, an emulsion foundation, a wax foundation and a spray, but the present invention is not limited thereto. The cosmetic composition of the present invention may also be formulated as a softening agent, a nutritional lotion, a nutritional 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.
<재료 및 방법>≪ Materials and methods >
본 실험에 사용된 천년초는 충남 연기군에서 생산된 것으로 네오크레마에서 제공하였다. 선인장의 점성 물질의 분해를 위하여는 Econitase CE(셀룰레이즈), Ultralflo L(베타-글루카네이즈), Cytolase PCL 5(펙티네이즈), Spezyme Prime(알파-아밀레이즈), Optidex L-400(글루코아밀레이즈), Pectinex 5XL, Celluclast 1.5L, optidex L-400, Rapidase(펙티네이즈, 헤미셀룰레이즈 및 셀룰레이즈) 및 Viscozyme(아라비네이즈, 셀룰레이즈, 베타-글루카네이즈, 헤미셀룰레이즈 및 자일레이즈)를 사용하였으며 이들은, 비전바이오캠으로부터 구입하였다(표 1).
The tsunami seeds used in this experiment were produced by Neo Crema, which was produced in Yeongi County, Chungnam Province. For the decomposition of viscous material of cactus, Econitase CE (Cellulase), Ultralflo L (Beta-Glucanase), Cytolase PCL 5 (Pectinase), Spezyme Prime (Alpha-Amylase), Optidex L-400 Pectinex 5XL, Celluclast 1.5L, Optidex L-400, Rapidase (pectinase, hemicellulase and cellulase) and Viscozyme (arabinose, cellulase, beta-glucanase, hemicellulase and xylase) Were purchased from Vision BioCam (Table 1).
효소의 특성Characteristics of enzymes
천년초Millennium 초고압 효소 추출 Extra-high pressure enzyme extraction
천년초의 줄기를 수세 후 잔가시를 제거한 다음 100 g에 물 500 ml을 가하여 homeogenizer 로 마쇄한 것을 효소 분해에 각각 사용하였다. 효소분해(효소 첨가량: 5 mL/100 g 천년초)는 표 1의 효소 특성에 따라 pH 5.0, 반응온도 50 ℃에서 60분간 가수분해하였으며, 이때 상압, 100, 200 및 300 MPa하에서 각각 분해를 진행하였다. 고압 효소 처리 후 효소 활성을 없애기 위해 끓는 물에 5분간 가열 처리 후 냉각 및 여과하여 그 상징액을 천년초 효소 가수분해액으로 사용하였다.
After washing the stem of the millennium, the residual sugar was removed and 100 g of water was added to 500 ml of water and homogenized with a homeogenizer. The enzymatic hydrolysis (enzyme addition amount: 5 mL / 100 g) was hydrolyzed at pH 5.0 and
천년초Millennium 분석방법 Analysis method
천년초 가수분해물의 폴리페놀함량은 Folin-Denis 법(Dewanto, V., Wu, X., & Liu, R. H.(2002b). Processed sweet corn has higher antioxidant activity. Journal of Agricultural and Food Chemistry, 50, 4959?4964.)에 준하여 측정하였으며, 총당과 산성당의 함량은 phenol-sulfuric acid법과 Blumenkrantz과 Asboe-Hansen법(Blumenkrantz, N. and Asboe-Hansen, G. (1973) Anal. Biochem., 54, 484)에 의해 측정하였다. The polyphenol content of the hydrolyzate of the millennia was measured by the Folin-Denis method (Dewanto, V., Wu, X., & Liu, RH (2002b), Processed sweet corn has higher antioxidant activity, Journal of Agricultural and Food Chemistry, 50, 4959? The contents of total sugars and acid sugars were determined by phenol-sulfuric acid method, Blumenkrantz and Asboe-Hansen method (Blumenkrantz, N. and Asboe-Hansen, G. (1973) Anal. Biochem., 54, 484) .
점도는 Brookfield viscometer(Model DV-II)를 이용하여 측정하였다. 이때 spindle No. 3을 사용하여 30 ℃에서 60 rpm의 조건에서 점도를 측정하였다.
Viscosity was measured using a Brookfield viscometer (Model DV-II). At this time, 3 was used to measure the viscosity at 30 DEG C and 60 rpm.
항산화 활성Antioxidant activity
항산화 활성은 ABTS와 DPPH라디칼 소거능에 의해 측정하였다. ABTS cationdecolorization assay법에 의해 시행하였다(Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 26: 1231-1237.). ABTS 7.4 mM과 potassium persulphate 2.6 mM을 하루 동안 암소에 방치하여 ABTS 라디칼을 형성시킨 후, 이 용액을 734 nm에서 흡광도 값이 1.0 이 되도록 몰 흡광계수(ε=3.6×104 M-1cm-1)를 이용하여 증류수로 희석하였다. 희석된 ABTS 라디칼 용액 1 mL에 추출액 50 ㎕를 가하여 흡광도의 변화를 정확히 90분 후에 측정하였다. ABTS 라디칼 제거능은 하기 식 1로 계산되었다.
Antioxidant activity was measured by ABTS and DPPH radical scavenging ability. ABTS cationdecolorization assay (Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. 1999. Free Radic Biol Med 26: 1231- 1237). After ABTS 7.4 mM and potassium persulphate 2.6 mM were incubated in the dark for one day to form ABTS radicals, the solution was incubated at 734 nm with a molar extinction coefficient (ε = 3.6 × 10 4 M -1 cm -1 ) Was diluted with distilled water. To 1 mL of the diluted ABTS radical solution, 50 μL of the extract was added and the change in absorbance was measured after 90 minutes. The ABTS radical scavenging ability was calculated by the following
<식 1><
ABTS 라디칼 소거능(%)＝(Ab-As)/Ab×100
ABTS radical scavenging ability (%) = (Ab-As) /
Ab: 추출물 대신 증류수를 넣었을 때의 흡광도 값의 변화Ab: Change in absorbance value when distilled water is added instead of extract
As: 추출물 넣었을 때의 흡광도 값
As: Absorbance value when put in extract
DPPH 라디칼 소거능에 의한 항산화활성은 Kim 등(Kim DO, Lee KW, Lee HJ, Lee CY. 2002. Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals. J Agric Food Chem 50: 3713-3717.)의 방법을 변형하여 실행하였다. 0.2 mM DPPH 용액 1 mL에 추출액 50 ㎕를 가하여 흡광도의 변화를 520 nm에서 정확히 30분 후에 측정하였다. DPPH 라디칼 제거능은 ABTS 라디칼 제거능과 동일한 식에 의해 계산하였다.
The antioxidant activity by DPPH radical scavenging was measured by the method of Kim et al. (Kim DO, Lee KW, Lee HJ, Lee CY 2002. Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals. J Agric Food Chem 50: 3713-3717. . 50 μl of the extract was added to 1 ml of 0.2 mM DPPH solution and the change in absorbance was measured at 520 nm exactly after 30 minutes. DPPH radical scavenging activity was calculated by the same equation as ABTS radical scavenging activity.
항보체Anti-complement 활성 측정 Active measurement
항보체 활성은 Mayer 방법을 이용하여 시료에 의한 보체 소비(complement consumption) 후 잔존하는 보체에 의한 적혈구 용혈 정도에 근거를 둔 complement fixation test로 측정하였다. 즉, 증류수에 녹인 시료에 정상인의 혈청 (normal human serum, NHS)과 GVB++(gelatin veronal buffered saline, pH 7.4, 2% gelatin, 500 μM Ca++, 2 mM Mg++ 함유) 완충액을 각각 50 ㎕씩 혼합하여 37 ℃에서 30분간 1차 반응시키고 GVB++를 350 ㎕를 가한 후 이를 10~160 배로 연속 희석하였다. 여기에 다시 750 ㎕의 GVB++를 가한 후 양의 감작적혈구(IgM-sensitization sheep erythrocyte, EA Cell, 1×108 cell/mL) 250 ㎕를 가해 37 ℃에서 60분간 2차 반응시키고 PBS 2.5 mL를 넣어 반응을 정지시켰다. 반응액을 2,500 rpm에서 약 10분간 원심분리 하여 얻어진 상등액을 412 nm에서 흡광도를 측정하여 잔존 용혈활성을 측정하였다. 항보체 활성은 NHS와 buffer, 증류수만을 반응시킨 대조군의 총보체 용혈(50% total complement hemolysis, TCH50)에 대한 저지율(inhibition of 50% total complement hemolysis, ITCH50, %)(식 2)로서 나타내었다.
Antibody activity was measured by the complement fixation test based on the degree of red blood cell hemolysis by the complement after consumption of complement by sample using the Mayer method. 50 μl of normal human serum (NHS) and GVB ++ (containing gelatin veronal buffered saline, pH 7.4, 2% gelatin, 500 μM Ca ++, and 2 mM Mg ++) were added to each sample dissolved in distilled water. For 30 min., 350 μl of GVB ++ was added thereto, and the resultant was serially diluted 10 to 160 times. After addition of 750 μl of GVB ++, 250 μl of positive sensitization sheep erythrocyte (EA Cell, 1 × 10 8 cells / mL) was added, followed by a secondary reaction at 37 ° C for 60 minutes and 2.5 mL of PBS The reaction was stopped. The reaction solution was centrifuged at 2,500 rpm for about 10 minutes, and the supernatant was measured for absorbance at 412 nm to measure the residual hemolytic activity. Anti-complement activity was expressed as inhibition of 50% total complement hemolysis (
<식 2><
ITCH50(%)={(대조군의 TCH50-샘플의 TCH50)/대조군의 TCH50}×100
ITCH 50 (%) = {(the control group TCH 50 - TCH 50 of the sample) / TCH 50 of the control group} × 100
<실험예 1> 효소 및 압력에 따른 천년초의 가수분해<Experimental Example 1> Hydrolysis of Milkwool with Enzymes and Pressure
전술한 방법에 따라 표 1의 효소를 이용하여 천년초를 상압 및 100 MPa 조건 하 가수분해하고 가수분해물의 라디칼 소거능 및 점도를 측정하였다. 이때, 대조군으로는 천년초 열수 추출물을 사용하였다.
According to the method described above, the enzyme of Table 1 was hydrolyzed at the atmospheric pressure and 100 MPa, and the radical scavenging ability and viscosity of the hydrolyzate were measured. At this time, as a control group, hot water extract of tsunami extract was used.
라디칼Radical 소거능Scatters
효소 추출물의 라디칼 소거능을 측정한 위한 결과, 상압(0.1 Mpa) 조건에서는 라피데이즈의 라디칼 소거능이 제일 우수하였고, 펙티넥스 처리군 역시 라디칼 소거능이 높았다.As a result of measuring the radical scavenging ability of the enzyme extract, the radical scavenging ability of lapidades was the best at the atmospheric pressure (0.1 Mpa) and the radical scavenging ability of the pectinex treated group was also high.
그러나 실험군 전체에서는 사이토레이즈 PCL 5를 100 MPa 조건 하 처리한 실험군의 DPPH 라디칼 소거능이 제일 우수하였으며, 펙티넥스를 100 MPa 조건 하 처리한 실험군의 라디칼 소거능이 그 다음으로 우수하였다(도 1). However, in the whole experimental group, the DPPH radical scavenging ability of the experimental group treated with
상압에서 높은 라디칼 소거능을 보였던 라피데이즈와 비스코자임는 100 MPa에서 라디칼 소거능이 감소하는 것으로 보아 압력에 의해 효소의 활성이 감소하는 것으로 추정된다. 반면, 100 MPa에서 라디칼 소거능이 우수하였던 사이토레이즈와 펙티넥스는 압력에 의한 효소활성감소가 적은 것으로 추정된다. 따라서 고압에서 천년초의 효소 처리에 적합한 것은 펙티넥스와 사이토레이즈였다.
The radical scavenging activity decreased at 100 MPa and the activity of enzyme was decreased by pressure due to the high radical scavenging ability at atmospheric pressure. On the other hand, cyto - raze and pectinex, which had excellent radical scavenging ability at 100 MPa, are presumed to have less enzyme activity due to pressure. Therefore, pectinex and cyto-raze were suitable for treatment of enzymes at high pressure.
점도Viscosity
상압 조건 하 실험군들의 점성 제거능은 라피데이즈 처리군이 가장 높았다. 비스코자임, 라피다제 처리군이 각각 4.87, 7.67 cP로 낮은 점도를 보였으나, 100 MPa처리시 9.56, 10.90 cP로 점성의 제거 효과가 낮았다. 반면, 상압에서 13.6과 13.8 cP의 낮은 점성 제거효과를 보였던 사이토레이즈와 펙티넥스는 100 MPa처리시 각각 6.4와 6.8 cP로 비교적 높은 점성 제거 효과를 보였다. 사이토레이즈 PCL 5를 100 MPa 조건 하 처리한 실험군의 점성 제거능이 가장 우수한 것으로 나타났다. (도 2). 그러므로 천년초 추출물이 우수한 라디칼 소거능을 가지면서도 천년초의 점성을 충분히 제거하는 방법은 천년초를 100 MPa에서 사이토레이즈 PCL 5 또는 펙티넥스로 처리하는 것으로 확인되었다.
The viscous removal capacity of the experimental groups was highest in the Rafidades treatment group at atmospheric pressure. Viscozyme and lapidase treatment showed low viscosity of 4.87 and 7.67 cP, respectively, but the removal effect of viscosity was low at 9.56 and 10.90 cP at 100 MPa treatment. On the other hand, CytoRase and Pectinex, which showed low viscosity of 13.6 and 13.8 cP at normal pressure, showed relatively high viscous removal effect at 6.4 and 6.8 cP at 100 MPa treatment, respectively. The experimental group treated with
<실험예 2><Experimental Example 2>
상기 실험예 1에서 우수한 점성 제거능 및 라디칼 소거능을 보인 펙티넥스 및 사이토레이즈 PCL 5를 하기 표 2의 중량비로 혼합하여 100 MPa 조건 하 천년초 마쇄물에 처리하였다. 그리고 상기 천년초 가수분해물의 라디칼 소거능을 측정하였다. 이때, 대조군으로는 천년초 열수 추출물을 사용하였다.
In Experimental Example 1, pectinex and
PC-1 내지 PC-5: 펙티넥스:사이토레이즈 PCL 5의 중량비PC-1 to PC-5: Pectinex: weight ratio of
대조군: 천년초 열수 추출물
Control group:
점도Viscosity
그 결과, 실험예 1에서 효소 단독 처리 시 100MPa에서 사이토레이즈 PCL 5와 펙테넥스를 혼합 사용할 경우 점도의 변화를 측정하였다. 펙티넥스와 사이토레이즈를 동량 홉합하였을 때(PC-3) 점도는 2.45 cP로 가장 낮은 값을 보였다(도 3). 그러므로 이하의 실험예 3 내지 8들은 사이토레이즈 PCL 5 및 펙티넥스를 동일한 중량으로 혼합한 효소 혼합물을 이용하여 수행하였다.
As a result, in Experimental Example 1, the viscosity change was measured when
<실험예 3> 천년초 초고압 추출물의 성분 분석≪ Experimental Example 3 >
사이토레이즈 PCL 5: 펙티넥스를 2:2로 포함하는 혼합 효소(이하, “혼합 효소”라 함)로 천년초를 압력에 따라 가수분해하고, 그 가수분해된 추출물의 dry weight, total sugar, uronic acid(산성당) 함량을 각각 측정하였다. 한편 비교군으로는 효소 처리 없이 초고압 추출을 수행한 천년초 초고압 추출물을 이용하였다. 각각의 실험군은 하기 표 3과 같으며, 이를 실험예 3 내지 실험예 6에서 사용하였다.
Total hydrolyzate of the hydrolyzed extract was measured by using a mixed enzyme (hereinafter referred to as " mixed enzyme ") containing 2: 2 cytoRease PCL 5: pectinex, (Acid group) content were measured. On the other hand, as the comparative group, ultra - Each experimental group is shown in Table 3 below and used in Experimental Examples 3 to 6.
*혼합 효소: 사이토레이즈 PCL 5: 펙티넥스를 2:2의 중량비로 혼합한 효소 혼합물
* Mixed enzyme: CytoRaze PCL 5: Enzyme mixture in which pectinex was mixed at a weight ratio of 2: 2
건물양Building quantity 및 폴리페놀 함량 And polyphenol content
천년초를 마쇄물의 경우(control) 18.2 mg/ml, 천년초 열수 추출물의 경우(Heat-ex) 21.9 mg/ml 의 건물양을 보였다. 또한 효소를 처리하지 않고 100 내지 300 MPa의 초고압에서 추출한 P-1군, P-2군 및 P-3군의 경우 각각 18.4, 19.0, 20.0 mg/ml의 건물양을 보였다. In the case of millennia, the amount of building was 18.2 mg / ml (control) and 21.9 mg / ml (heat-ex) in the case of hot-water extract from the tiller. In the P-1 group, P-2 group and P-3 group extracted at an ultra-high pressure of 100 to 300 MPa without the enzyme treatment, the amount of building was 18.4, 19.0 and 20.0 mg / ml, respectively.
한편, 초고압과 혼합 효소 처리를 병용하였을 경우 P-1-E군, P-2-E군 및 P-3-E군은 각각 35.3, 35.0과 36.8 mg/ml의 건물량을 보였는바, 초고압과 효소를 병용 처리하였을 경우 추출되어 회수되는 건물의 양이 증가하는 것으로 확인되었다.
On the other hand, P-1-E, P-2-E, and P-3-E groups showed 35.3, 35.0 and 36.8 mg / ml dry weight, respectively, It was confirmed that the amount of extracted and recovered buildings increased when the enzyme was used in combination.
또한 추출물중의 활성성분으로 여겨지는 폴리페놀의 함량 역시 초고압 효소추출의 경우 P-1-E군, P-2-E군 및 P-3-E군은 각각 0.25, 0.27 및 0.28 mg/ml로 control(0.18 mg/ml)과 Heat-ex군(0.19 mg/ml)에 비하여 높은 함량을 보였다. 또한, 100 내지 300MPa의 효소 무처리 초고압 추출물들(0.17 내지 0.18 mg/ml)에 비하여도 높은 함량을 보였다(도 4). The content of polyphenols, which are considered to be active components in the extracts, was 0.25, 0.27 and 0.28 mg / ml in the P-1-E, P-2-E and P- (0.18 mg / ml) and Heat-ex group (0.19 mg / ml), respectively. In addition, the content was higher than that of the enzyme-free ultra-high pressure extracts (0.17-0.18 mg / ml) of 100-300 MPa (FIG. 4).
그러나 혼합 효소 처리 초고압 추출물 간 100, 200, 300 MPa의 압력의 차이는 없었다.
However, there was no difference in pressure between 100, 200, and 300 MPa between the high - pressure extracts treated with mixed enzyme.
총 당Per capita 및 산성당 And Sanctuary
초고압 처리시 건물량, 폴리페놀과 더불어 천년초의 가수분해 정도를 판단할 수 있는 지표가 총 당(total sugar) 및 산성당(uronic acid)의 함량 변화이다. 총 당 함량 변화 역시 효소 처리 없이 초고압만 처리하는 경우보다 혼합 효소와 병용 처리시 증가하는 경향을 보였으며, 면역활성 증진 물질로 알려진 uronic acid 함량 역시 초고압/효소 처리시 증가하는 경향을 보였다.
Changes in total sugar and uronic acid content are indicators of the amount of dry matter, polyphenol, and hydrolysis rate in millennials at ultra high pressure. Changes in total sugar content also increased with the mixed enzyme treatment, compared with the case without the enzyme treatment, and the uronic acid content, also known as the immunoreactivity enhancing substance, also tended to increase with the high pressure / enzyme treatment.
초고압 혼합 효소 추출물의 경우 P-1-E군, P-2-E군 및 P-3-E군은 총당과 산성당의 함량이 각각 17.27, 16.53, 18.94 mg/ml과 3.44, 3.63, 3.53 mg/ml의 함량으로 control (9.61, 0.84 mg/ml)과 heat-ex군(11.40, 1.25 mg/ml)보다 높은 함량을 보였다. 건물량과 폴리페놀 함량과 마찬가지로 효소 처리 없이 단순히 초고압 추출한 경우(9.44-10.32 mg/ml, 0.81-1.12 mg/ml)에 비하여 높은 함량을 갖는 것으로 나타났다(도 5). 이상의 결과에 의해, 천년초를 초고압/효소 병용처리하는 것이 유용성분의 증대 효과를 기대할 수 있는 것으로 확인되었다.
In the case of P-1-E, P-2-E and P-3-E, the total sugar and acidic sugar contents were 17.27, 16.53 and 18.94 mg / ml and 3.44, 3.63 and 3.53 mg / ml in the control group (9.61, 0.84 mg / ml) and heat-ex group (11.40, 1.25 mg / ml). Similar to the dry matter and polyphenol contents, it was found to be higher than the case of simply extracting with ultra high pressure (9.44-10.32 mg / ml, 0.81-1.12 mg / ml) without enzyme treatment (Fig. 5). From the above results, it was confirmed that the treatment of the combination of the high-pressure / enzyme treatment of the millennia early can expect the effect of increasing the useful ingredient.
<실험예 4> 천년초 초고압 추출물의 라디칼 소거능<Experimental Example 4> Radical scavenging ability of ultra high pressure extract
천년초 추출물의 라디칼 소거능을 측정하고자 IC50값을 구하여 각각의 활성을 비교하였다. 천년초 마쇄물(control), 열수 추출물(Heat-ex군)의 경우 DPPH와 ABTS에 대한 IC50 값은 각각 4.11, 0.63 mg/ml, 2.20, 0.44 mg/ml의 활성을 보인 반면 초고압과 혼합 효소를 병용 처리한 P-1-E군, P-2-E군 및 P-3-E군은 각각 0.62, 0.11 mg/ml, 0.55, 0.13 mg/ml, 0.47, 0.12 mg/ml로 낮은 IC50값을 보였는바, 라디칼 소거능이 우수한 것으로 나타났다(도 6).
In order to measure the radical scavenging ability of the extracts, IC50 values were determined and the activities of the extracts were compared. IC50 values for DPPH and ABTS were 4.11, 0.63 mg / ml, 2.20 and 0.44 mg / ml, respectively, in the case of control and hot-water extracts. The IC50 values of the treated P-1-E, P-2-E and P-3-E groups were as low as 0.62, 0.11 mg / ml, 0.55 and 0.13 mg / ml, 0.47 and 0.12 mg / Bar, and radical scavenging ability (FIG. 6).
특히, 초고압 효소 병용처리시 DPPH에 대한 소거능은 P-3-E군이 높은 반면, ABTS에 대한 소거능은 P-3-E군에 비하여 P-1-E군이 다소 높은 라디칼 소거능을 보였다.
Particularly, the P-3-E group showed higher scavenging ability to DPPH than the P-3-E group, while the scavenging ability to ABTS showed a higher radical scavenging ability than the P-3-E group.
<실험예 5> 천년초 초고압 추출물의 항보체 활성≪ Experimental Example 5 >
면역활성의 비교를 위하여 각 추출 방법에 따른 항보체 활성을 측정하였다. 그 항보체 활성의 기준물질인 LPS(lipopolysaccharide)의 활성은 1000 ㎍/ml에서 60%의 항보체 활성을 보인 반면, 마쇄물(control) 및 열수 추출물(Heat-ex군)은 1000 ㎍/ml에서 각각 15.0%와 27.4%의 항보체 활성을 보인 반면, 초고압 조건 하 무효소 추출한 P-1군, P-2군 및 P-3군은 1000 ㎍/ml에서 각각 34.2%, 33.5%, 31.3%의 항보체 할성을 보였다. 반면 초고압 효소 추출물인 P-1-E군, P-2-E군 및 P-E-3군은 1000 ㎍/ml에서 각각 48.9%, 50.2%, 51.2%의 항보체 활성을 보였다(도 7). 초고압 효소 추출은 다른 추출방법에 비하여 높은 항보체 활성을 보였으며, 초고압 추출시 적용하였던 100 MPa, 200 MPa, 300 MPa의 압력간의 유의적인 차이는 없었다.
For the comparison of immunological activity, anti - complement activity according to each extraction method was measured. The activity of LPS (lipopolysaccharide), a reference substance for its anti-complement activity, showed 60% anti-complement activity at 1000 μg / ml, whereas control and hot-water extract (Heat-ex group) , P-1, P-2, and P-3 groups, respectively, showed an anticoagulant activity of 15.0% and 27.4% Showed anti - complementation ability. On the other hand, the P-1-E, P-2-E and PE-3 hypervariable enzyme extracts showed anti-complement activity of 48.9%, 50.2% and 51.2% at 1000 ㎍ / ml, respectively (FIG. The ultrafiltration enzyme activity showed higher antioxidant activity than the other extraction methods, and there was no significant difference between the pressures of 100 MPa, 200 MPa and 300 MPa applied in ultra high pressure extraction.
<실험예 6> 천년초 초고압 추출물의 점도<Experimental Example 6> Viscosity of the ultra-high pressure extract
각 추출 방법에 따른 점도의 변화를 측정한 결과, control인 천년초 마쇄 추출물의 경우 40.9 cP의 점도을 보였으며, 열수 추출물의 경우 이보다 낮은 13.2 cP점도를 보였다. 무효소 초고압 추출물인 P-1군, P-2군 및 P-3군은 각각 21.2, 20.3, 19.8 cP의 점도로 열수추출물에 비하여 다소 높은 점도를 보인 반면, 초고압 효소 추출물인 P-1-E군, P-2-E군 및 P-3-E군은 각각 3.4, 2.8과 2.7 cP로 낮은 점도를 보였다(도 8).
As a result of measuring the change of viscosity according to each extraction method, the viscosity of the control granulated pollen seed extract was 40.9 cP, and that of the hot water extract was 13.2 cP. The P-1 group, P-2 group and P-3 group showed higher viscosity than the hot-water extracts at 21.2, 20.3 and 19.8 cP, respectively, while the P-1-E Group, P-2-E and P-3-E groups showed low viscosities of 3.4, 2.8 and 2.7 cP, respectively (Fig. 8).
이상의 결과에 의하면 초고압 효소 추출은 다른 추출방법에 비하여 효과적인 추출방법임을 확인하였으며, 초고압 효소 추출의 경우 100, 200, 300 MPa의 각 압력간의 차이는 유의적이지 않았다. 따라서 비용 대비 가수분해 효율을 고려할 때, 100 MPa에서 효소를 처리하는 방법이 효율적일 것이다.
From the above results, it was confirmed that the extraction method of ultrahigh pressure enzyme was an effective extraction method compared with other extraction methods. In the case of ultra high pressure enzyme extraction, the difference between the pressures of 100, 200 and 300 MPa was not significant. Therefore, considering the cost-effectiveness of hydrolysis, it would be efficient to treat the enzyme at 100 MPa.
<실험예 7> 추출 시간에 따른 성분 변화≪ Experimental Example 7 >
상기 실험예 3 내지 6에서 가장 효율적으로 판단된 조건인 100 MPa 압력 하 천년초를 혼합 효소로 추출 시, 추출 시간에 따른 추출물 내 성분 변화를 측정하였다.
In the experiment examples 3 to 6, when the extract was extracted with the mixed enzyme at a pressure of 100 MPa, which was the most efficient condition, the change in the extract components was measured according to the extraction time.
그 결과, 추출시간이 증가할수록 총당, 산성당 및 폴리페놀 함량이 증가하는 경향을 보였으며, 점도는 추출시간이 증가할수록 감소하는 경향을 보였다. 점도의 경우 초기 40.9 cP였으나 1, 2, 4시간 추출 시 각각 33.4, 2.7과 2.5로 감소하였다(도 9a).As a result, total sugar, acid sugar and polyphenol content tended to increase with increasing extraction time, and viscosity tended to decrease with increasing extraction time. The initial viscosity was 40.9 cP, but it decreased to 33.4, 2.7 and 2.5 at 1, 2 and 4 hours extraction, respectively (Fig. 9a).
총당 및 산성당 역시 초기에는 각각 9.6 mg/ml과 0.84 mg/ml로 낮은 함량을 보였다. 그러나 1시간, 2시간 및 4시간 효소 추출 시, 총당 함량은 각각 17.27, 18.9, 19.3 mg/ml의 변화를 보였고, 산성당은 3.44, 3.67, 3.70 mg/ml로 서서히 증가하는 경향을 보였다(도 9b). 폴리페놀 함량 역시 효소 추출시간이 증가할수록 초기 0.18 mg/ml에서 1시간, 2시간 및 4시간 추출 시 각각 0.25, 0.35, 0.38 mg/ml의 함량 변화를 보였다(도 9c)Total sugar and acid were also initially low at 9.6 mg / ml and 0.84 mg / ml, respectively. However, when 1, 2, and 4 hours of enzyme digestion were performed, the total sugar content was 17.27, 18.9, and 19.3 mg / ml, respectively, and acidity was gradually increased to 3.44, 3.67, and 3.70 mg / ml 9b). The content of polyphenol contents was 0.25, 0.35, and 0.38 mg / ml at 1 hour, 2 hours and 4 hours, respectively, at the initial 0.18 mg / ml as the enzyme extraction time increased (FIG. 9c)
효소 추출시간에 따른 성분 간의 함량 변화는 2시간 까지 급격히 변화가 진행되다가 그 시간 이후 서서히 증가(점도의 경우는 감소)하는 경향을 보였다.
The contents of the components varied with the time of enzyme extraction and showed a tendency to increase rapidly after 2 hours and gradually increase (decrease in viscosity) after that time.
<실험예 8> 추출 시간에 따른 라디칼 소거능의 변화<Experimental Example 8> Changes in radical scavenging ability according to extraction time
상기 실험예 3 내지 6에서 가장 효율적으로 판단된 조건인 100 MPa 압력 하 천년초를 혼합 효소로 추출 시, 추출 시간에 따른 추출물의 라디칼 소거능 차이를 측정하였다.
The radical scavenging ability of the extract was measured according to the extraction time when the mixed enzyme was extracted under the most effective condition of 100 MPa pressure in Experimental Examples 3 to 6.
초고압 효소 추출시 시간에 따른 DPPH 및 ABTS 라디칼 소거능을 측정한 결과, 라디칼 소거능을 나타내는 IC50값의 변화는 추출시간이 증가할수록 증가하는 경향을 보였으나, 추출 2시간 이후의 변화는 미미하였다.
As a result of measuring DPPH and ABTS radical scavenging activity over time in the extraction of ultrahigh pressure enzyme, the IC50 value indicating the radical scavenging ability tended to increase with increasing extraction time, but the change after 2 hours of extraction was insignificant.
ABTS 라디칼 소거능은 추출 전 630.2 ㎍/ml의 IC50값을 보였으나 추출 후 1시간이 경과하자 110.3 ㎍/ml으로 현저히 증가하였다. 그러나 2시간 및 4 시간 추출 시 각각 95.7 및 95.4 ㎍/ml으로 2시간 이후 활성 변화가 미미하였다. 한편, DPPH 라디칼 소거능 역시 추출 전 4100.3 ㎍/ml이었으나, 추출 후 1시간이 경과하자 620.4 ㎍/ml으로 현저히 증가하였고, 2시간 및 4시간 추출 시 각각 590.3 및 586.2 ㎍/ml으로 1시간 이후 활성의 변화가 미미 하였다(도 10).
The ABTS radical scavenging activity was 630.2 ㎍ / ml before extraction but increased to 110.3 ㎍ / ml after 1 hour of extraction. However, 95% and 95.4 ㎍ / ml of 2 hour and 4 hour extraction, respectively, showed little activity change after 2 hours. The DPPH radical scavenging activity was also 4100.3 ㎍ / ml before extraction but increased to 620.4 ㎍ / ml after 1 hour of extraction. The activity of DPPH radical scavenging activity was increased to 590.3 and 586.2 ㎍ / ml, respectively, The change was minimal (Fig. 10).
그러므로 추출 시간 대비 라디칼 소거능을 고려할 때, 1시간 내지 2시간 동안 추출하는 것이 가장 효율적일 것으로 판단되었다. 이는 추출 시간 대비 점도 변화, 추출물 내 성분 변화의 효율과도 일치하는 것으로, 점도, 유효 성분 및 라디칼 소거능을 모두 고려 시, 초고압 조건 하 1시간 내지 2시간 동안 혼합 효소로 천년초를 추출하는 것이 가장 효율적이다.
Therefore, when the radical scavenging ability is compared with the extraction time, extraction for 1 to 2 hours was considered to be most effective. Considering both the viscosity, the active ingredient and the radical scavenging ability, it is most efficient to extract the silkworms with the mixed enzyme for 1 hour to 2 hours under the super high pressure condition, to be.
Claims (10)
A method for producing a Chinese medicinal herb extract comprising the step of hydrolyzing and hydrolyzing at least one enzyme selected from the group consisting of pectinex and cytoRease PCL 5 under the ultrahigh pressure condition to the Opuntia humifusa to remove the viscosity of the perennial plant.
100MPa ~ 1,000MPa의 압력 하 효소 처리를 수행하는 것을 특징으로 하는 제조 방법.
The method according to claim 1,
Characterized in that an enzyme treatment is carried out under pressure of 100 MPa to 1,000 MPa.
100MPa~300MPa의 압력 하 효소 처리를 수행하는 것을 특징으로 하는 제조 방법.
The method according to claim 1,
Characterized in that an enzyme treatment is carried out under a pressure of 100 MPa to 300 MPa.
100MPa~200MPa의 압력 하 효소 처리를 수행하는 것을 특징으로 하는 제조 방법.The method according to claim 1,
Characterized in that an enzyme treatment is carried out under a pressure of 100 MPa to 200 MPa.
상기 효소는 펙티넥스 및 사이토레이즈 PCL 5의 혼합물인 것을 특징으로 하는 제조 방법.
The method according to claim 1,
Wherein said enzyme is a mixture of pectinex and cytoRase PCL 5.
상기 효소는 펙티넥스 및 사이토레이즈 PCL 5를 3:1 내지 1:3의 중량비로 혼합한 혼합물인 것을 특징으로 하는 제조 방법.
The method according to claim 1,
Wherein the enzyme is a mixture of pectinex and cytoRase PCL 5 in a weight ratio of 3: 1 to 1: 3.
상기 천년초 추출물은 DPPH 라디칼 소거능 및 15 cP 이하의 점도를 갖는 것을 특징으로 하는 제조 방법.
The method according to claim 1,
Wherein the citrus fruit extract has a DPPH radical scavenging activity and a viscosity of 15 cP or less.
8. A cosmetic composition comprising an effective amount of a tsunami extract prepared by the method of any one of claims 1 to 7.
8. A food composition comprising as an active ingredient an extract of a Korean lilacs produced by the method of any one of claims 1 to 7.
And hydrolyzing at least one enzyme selected from the group consisting of pectinex and cytoResize PCL 5 to hydrolysis in Opuntia humifusa under ultra high pressure conditions.
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