KR101063351B1 - The extracts and fractions of Hippophae rhamnoides L. - Google Patents
The extracts and fractions of Hippophae rhamnoides L. Download PDFInfo
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- KR101063351B1 KR101063351B1 KR1020100031410A KR20100031410A KR101063351B1 KR 101063351 B1 KR101063351 B1 KR 101063351B1 KR 1020100031410 A KR1020100031410 A KR 1020100031410A KR 20100031410 A KR20100031410 A KR 20100031410A KR 101063351 B1 KR101063351 B1 KR 101063351B1
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- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000007218 ym medium Substances 0.000 description 1
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 1
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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Abstract
본 발명은 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화제, 항생제 및 항당뇨용 조성물에 관한 것으로, 더욱 상세하게는 산자나무를 유기용매로 추출하여 제조되는 추출물 또는 이의 분획물을 포함하는 항산화제, 항생제 및 항당뇨용 조성물에 관한 것이다. 본 발명의 산자나무 추출물 또는 이의 분획물은 항산화 효과를 나타냄으로써 항산화 또는 노화방지 화장품 또는 약학적 조성물로 유용하게 이용될 수 있고, 항균 및 항진균 효과를 나타냄으로써 항생제 또는 건강식품으로 유용하게 이용될 수 있고, 항당뇨 효과를 나타냄으로써 항당뇨용 조성물 또는 건강식품으로 유용하게 이용될 수 있다.The present invention relates to an antioxidant, an antibiotic and an antidiabetic composition comprising an extract of Ornithiasis japonica or a fraction thereof as an active ingredient. More particularly, the present invention relates to an antioxidant, an antioxidant, Antibiotics, and antidiabetic agents. INDUSTRIAL APPLICABILITY The extract of Wakamatsu or its fractions of the present invention exhibits an antioxidative effect and thus can be effectively used as an antioxidant or antioxidant cosmetic or pharmaceutical composition and exhibits antimicrobial and antifungal effects, thus being useful as an antibiotic or a health food , And exhibit an antidiabetic effect, so that it can be usefully used as a composition for antidiabetic use or as a health food.
Description
본 발명은 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화제, 항생제 및 항당뇨용 조성물에 관한 것이다.The present invention relates to an antioxidant, an antibiotic and a composition for antidiabetic use which comprise an extract of Liliaceae or a fraction thereof as an active ingredient.
최근 소득 증대로 인하여 개인의 영양상태가 크게 호전되었고 의료 기술의 발달로 평균 수명이 선진국 수준으로 많이 증가하고 있다. 이에 따라 질병의 치료보다는 질병 예방에 힘써야 한다는 생각에 여러 형태의 건강 기능성 식품에 대한 수요에 초점을 맞추고 있다. 질병을 예방하기 위한 생체 방어시스템으로 천연물을 대상으로 하는 연구가 활발히 수행되고 있으며, 천연물의 기능성과 천연물에 함유하고 있는 2차 대사산물의 생리활성 효과에 대한 연구가 진행되고 있다. 또한 천연식물로 노화와 성인병 질환의 원인인 생체 내에서 발생하는 산화적 스트레스의 직접적 원인이 되는 활성산소 종(reactive oxygen species : ROS)을 소거할 수 있으며 식품에 존재하는 항산화 물질들이 과산화물에 의해 야기되는 각종 노화성질환의 억제제 및 치료제로 개발될 수 있다고 기대되는 바이다.The recent increase in income has greatly improved the nutritional status of the individual and the average life expectancy has increased to the level of advanced countries due to the development of medical technology. Therefore, we focus on the demand for various types of health functional foods in order to prevent diseases and to prevent diseases. Studies on natural products have been actively carried out as biological defense systems to prevent diseases. Studies on the functionality of natural products and the effects of the secondary metabolites contained in natural products on physiological activity are under way. In addition, it is possible to eradicate reactive oxygen species (ROS) which is a direct cause of oxidative stress in the living body, which is a cause of aging and diseases of natural diseases, and the antioxidant substances present in foods are caused by peroxides It is expected to be developed as an inhibitor and therapeutic agent for various aging diseases.
산자나무(Hippophae rhamnoides L.; 비타민나무)는 보리수과에 속하는 관목으로서 100 여종 이상의 성분을 포함하고 있으며, 특히 비타민(A, B, C, E, F, K)등 유효한 성분을 많이 함유하고 있다. 겨울철 추위 및 여름철 고온에서도 잘 자라며, 척박한 지대에서도 잘 자라는 강한 적응성을 지녔다. 이미 산자나무의 항산화 활성 측정 및 산자나무의 지방산 함량 및 암 예방, 면역 체계 분야 등 다양한 연구 분야가 이루어지고 있으며 산자나무 열매에 베타 카로틴 성분이 함유하고 있다고 보고된 바 있다. 또한 산자나무 잎의 기기분석을 통해 유효성분을 측정하고 있으며, 산자나무로 다양한 식품을 만들어 사용하고 있으나, 국내에서는 아직 그 연구가 활발하게 이루어지지 않고 있다. Hippophae rhamnoides L .; Vitamin tree) is a shrub belonging to the bloom tree and contains more than 100 kinds of ingredients. Especially, it contains a lot of effective ingredients such as vitamins (A, B, C, E, F and K). It grows well in the cold of winter and the high temperatures of summer, and has a strong adaptability that grows well in the barren zone. It has already been reported that antioxidant activity of Sanjia wood is investigated, and fatty acid content, cancer prevention, and immune system of Sanjay tree are being studied. Beta carotene is contained in the fruit tree. In addition, the active ingredient is measured by instrumental analysis of Sanjia leaves, and various foods are made using Sanjia wood, but the research is not active in Korea yet.
이에 본 발명자들은 산자나무 뿌리 및 줄기 추출물 및 이의 분획물의 전자공여능, SOD 유사 활성, 항균 및 항진균 활성, 3가철 활성(Ferric-thiocyanate activity), α-글루코시다아제 활성을 측정하고, 본 발명의 추출물 및 이의 분획물이 항산화제, 항생제 및 항당뇨용 조성물로써 사용될 수 있음을 확인함으로써 본 발명을 완성하였다.
Therefore, the present inventors measured the electron donating ability, SOD-like activity, antibacterial and antifungal activity, ferric-thiocyanate activity and? -Glucosidase activity of the extracts of the root and stem extracts and fractions thereof, And fractions thereof can be used as antioxidants, antibiotics and antidiabetic compositions.
본 발명의 목적은 항산화 활성을 나타내는 산자나무 추출물 또는 이의 분획물을 제공하는 것이다.It is an object of the present invention to provide an extract of Ornithiasis or its fraction showing antioxidative activity.
또한, 본 발명의 목적은 항균 및 항진균 활성을 나타내는 산자나무 추출물 또는 이의 분획물을 제공하는 것이다.It is also an object of the present invention to provide a saponin extract or fractions thereof exhibiting antibacterial and antifungal activity.
아울러, 본 발명의 목적은 항당뇨 활성을 나타내는 산자나무 추출물 또는 이의 분획물을 제공하는 것이다.In addition, the object of the present invention is to provide a saponin extract exhibiting an anti-diabetic activity or a fraction thereof.
상기 목적을 달성하기 위하여, 본 발명은 물, 알코올 또는 이들의 혼합물로 추출되는 산자나무(Hippophae rhamnoides L.; 비타민나무) 추출물을 제공한다.In order to achieve the above object, the present invention relates to a method for producing a plant extract of Hippophae rhamnoides L .; Vitamin tree) extracts.
또한, 본 발명은 상기 산자나무 추출물을 n-헥산, 에틸아세테이트 및 n-부탄올 순으로 추출하여 각 단계에서 얻은 분획물을 제공한다.In addition, the present invention provides fractions obtained in each step by extracting the above-mentioned extract from the above-mentioned extracts in the order of n-hexane, ethyl acetate and n-butanol.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화제를 제공한다.In addition, the present invention provides an antioxidant comprising the above-described extract of Sanjoboya or fractions thereof as an active ingredient.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 활성산소가 과량으로 축적되어 발병되는 질환의 예방 및 치료용 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing and treating diseases in which excessive amounts of active oxygen containing an extract of the Acanthopanthis mothera or its fractions as an active ingredient are accumulated and developed.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 함유하는 노화방지용 약학적 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing aging which contains the above-described extract of Liliaceae or its fractions as an active ingredient.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 포함하는 피부 노화 방지용 화장품을 제공한다.In addition, the present invention provides a cosmetic for preventing skin aging comprising the above-mentioned extract of Liliaceae or its fractions.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항생제를 제공한다.In addition, the present invention provides an antibiotic comprising the above extract of Sanjobo or fractions thereof as an active ingredient.
또한, 본 발명은 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항당뇨용 조성물을 제공한다.In addition, the present invention provides a composition for anti-diabetic comprising an extract of Ornithiasis or its fraction as an active ingredient.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 천연방부제를 제공한다.In addition, the present invention provides a natural preservative comprising the above-described extract of Sanjay or fractions thereof as an active ingredient.
아울러, 본 발명은 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 당뇨병 예방 및 개선용 건강식품을 제공한다.
In addition, the present invention provides a health food for prevention and improvement of diabetes mellitus comprising an extract of Ornithiasis or its fraction as an active ingredient.
이하, 본 발명을 상세히 설명한다.
Hereinafter, the present invention will be described in detail.
본 발명은 산자나무(Hippophae rhamnoides L.) 추출물을 제공한다.The present invention relates to a method for the production of Hippophae rhamnoides L.) extract.
상기 산자나무는 재배한 것 또는 시판되는 것 등 제한 없이 사용할 수 있다. 또한, 본 발명에 따른 산자나무 추출물은 다음과 같이 제조될 수 있다. 산자나무 뿌리 및 줄기를 물로 깨끗이 세척한 후, 그늘 및 실온에서 7일간 건조한다. 건조한 산자나무 뿌리 및 줄기를 분쇄한 후, 추출 용기에 넣고, 추출용매로 적절한 온도에서 일정 시간 추출한다. 상기 산자나무 추출물을 수득한 후, 거름종이 등을 이용하여 고형분을 제거하고, 현탁액을 원심분리시키고, 상층액을 감압 여과할 수 있다. 상기 추출 용매는 물, 알코올 또는 이의 혼합물, 바람직하게는 C1 내지 C4의 저급 알코올 또는 이들의 혼합 용매로부터 선택된 용매를 사용하는 것이 바람직하며, 메탄올을 사용하는 것이 더욱 바람직하나, 이에 한정되는 것은 아니다. 상기 추출 용매의 양은 산자나무 건조 중량의 2 내지 10 배로, 바람직하게는 4배로 한다. 상기 추출 방법은 침지 추출, 열 추출, 환류 냉각 추출 및 초음파 추출 등의 추출 방법을 사용할 수 있으며, 바람직하게는 침지 추출방법으로 1회 내지 5회 추출될 수 있다. 추출시 온도는 20℃ 내지 30℃ 이며, 더욱 바람직하게는 20℃ 내지 25℃인 실온에서 수행하는 것이 바람직하다. 상기 추출 시간은 5 내지 10일이며, 바람직하게는 7일이다.
The above tree can be used without limitation such as cultivated or marketed. In addition, the extract of Liliaceae according to the present invention can be prepared as follows. After washing the roots and stems of the plants with water, they are dried in the shade and room temperature for 7 days. After the dry roots and stems are crushed, they are placed in an extraction vessel and extracted with an extraction solvent at a suitable temperature for a certain period of time. After obtaining the above-mentioned extract, the solid content is removed using a filter paper or the like, the suspension is centrifuged, and the supernatant can be filtrated under reduced pressure. The extraction solvent is preferably a solvent selected from water, an alcohol or a mixture thereof, preferably a C 1 to C 4 lower alcohol or a mixed solvent thereof. More preferably, methanol is used. However, no. The amount of the extraction solvent is 2 to 10 times, preferably 4 times, the dry weight of Sanjay tree. The extraction method may be an extraction method such as an immersion extraction method, a heat extraction method, a reflux cooling extraction method, and an ultrasonic extraction method. Preferably, the extraction method may be performed once to five times by an immersion extraction method. The temperature at the time of extraction is preferably 20 ° C to 30 ° C, more preferably 20 ° C to 25 ° C at room temperature. The extraction time is 5 to 10 days, preferably 7 days.
또한, 본 발명은 상기 산자나무 추출물을 n-헥산, 에틸아세테이트 및 n-부탄올 순으로 추출하여 각 단계에서 얻은 분획물을 제공한다.In addition, the present invention provides fractions obtained in each step by extracting the above-mentioned extract from the above-mentioned extracts in the order of n-hexane, ethyl acetate and n-butanol.
상기 산자나무 추출물의 분획물을 획득하기 위한 분획용매는 유기용매이고, 바람직하게는 n-헥산, 에틸아세테이트 및 n-부탄올 군에서 선택되며, 통상적으로 분별깔때기를 이용하여 분획하였다. 상기 분획물은 상기 추출물로부터 분획 과정을 1 내지 5회, 바람직하게는 3회 반복하여 수득할 수 있고, 분획 후 감압 농축될 수 있다.The fraction solvent for obtaining fractions of the above extract is selected from the group consisting of n-hexane, ethyl acetate and n-butanol, and is usually fractionated using a separatory funnel. The fraction may be obtained from the extract by repeating the fractionation process one to five times, preferably three times, and may be fractionated and then concentrated under reduced pressure.
본 발명의 실시예에서는 상기 산자나무 추출물을 증류수에 현탁시킨 후, n-헥산, 에틸아세테이트 및 n-부탄올 순으로 계통 분획한 후, 이를 3회 반복하고 각 용매 분획물을 감압 농축하였다.
In the example of the present invention, the above-described extract of the bitter gourd was suspended in distilled water, followed by fractionation in the order of n-hexane, ethyl acetate and n-butanol. The fraction was repeated three times, and the solvent fractions were concentrated under reduced pressure.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화제를 제공한다.In addition, the present invention provides an antioxidant comprising the above-described extract of Sanjoboya or fractions thereof as an active ingredient.
전자공여능은 항산화작용의 지표이며, 환원력이 큰 것일수록 전자공여능이 높다고 알려져 있다(Kang YH et al ., Korean J Food Sci Technol 27:978-984, 1995). 이에 상기 산자나무 추출물 또는 이의 분획물의 항산화 활성을 기존의 항산화제인 BHA, BHT, α-토코페롤과 비교한 결과, 상기 대조군에 비해 높은 전자공여능을 나타냈으며, 특히 뿌리 및 줄기의 EtOAc(에틸 아세테이트) 분획에서 각각 RC50이 1.5 ± 0.5 ㎍/㎖ 및 1.0 ± 0.5 ㎍/㎖로 이는 대조군 α-토코페롤 RC50 12 ± 0.5 ㎍/㎖과 비교해 볼 때 더 강한 전자공여능을 나타냈다(표 1 참조).The electron donating ability is an index of antioxidant activity, and it is known that the electron donating ability is higher as the reducing power is larger (Kang YH meat al . , Korean J Food Sci Technol 27: 978-984,1995). The antioxidative activity of the extracts of the above-mentioned extracts or fractions thereof was compared with those of the existing antioxidants BHA, BHT and? -Tocopherol. As a result, the antioxidative activity of the extracts was higher than that of the control group. Particularly, EtOAc (ethyl acetate) RC 50 was 1.5 ± 0.5 ㎍ / ㎖ and 1.0 ± 0.5 ㎍ / ㎖, respectively, which showed stronger electron donating ability compared to the control α-tocopherol RC 50 12 ± 0.5 ㎍ / ㎖ (see Table 1).
또한, 체내에서 활성산소 라디칼을 산화시켜주는 효소로 산소 상해로부터 생체를 보호하는 기능이 있는 것으로 알려져 있는 활성산소제거효소(Superoxide dismutase; SOD) 유사 활성(like activity) 작용은 산자나무에서 메탄올 추출물을 비롯한 모든 분획물에서 농도가 증가할수록 농도 의존적으로 증가하였다(표 2 참조). 산자나무 추출물 및 이의 분획물은 각각 저농도에서는 활성이 큰 차이를 보이지 않았으나, 산자나무 헥산 분획물 고농도(1,000 ppm)에서는 각각 65.7% 또는 73.4% 억제율을 나타났다(표 2 참조). 상기 결과는 Hong HD 등(Korean J Food Sci Technol 30:1484-1487, 1998)의 과실, 과채류의 착즙의 SOD 유사 활성에서 사과 착즙액의 경우 14.6%, 케일농축액의 경우 26.7%, 키위 착즙액의 경우 27.6%, 무착즙액의 경우 24.1%의 활성을 나타낸 것에 비하여 높은 SOD 유사 활성을 나타내었다. 또한, 상기 결과는 폴리페놀과 플라보노이드의 함량이 높을수록 SOD의 유사 활성이 증가한다는 보고(Kwon TD et al ., Kor J Phys Edu 3:891-899, 2001)와 기존의 연구결과(Azuma K et al ., L J Agric Food Chem 47:3963-3966, 1999; Lee YS et al ., Korean J Food Preserv 12:75-79, 2005)에 근거하여 산자나무의 뿌리와 줄기에 폴리페놀이나 플라보노이드를 포함한 생리활성 물질을 많이 함유하고 있는 것으로 생각되므로 이를 기능성 제품에 활용할 수 있을 것으로 사료된다.In addition, superoxide dismutase (SOD) -like activity, which is known to have the function of protecting the organism from oxygen injury by an enzyme that oxidizes the active oxygen radical in the body, (See Table 2). In the low concentration of the extracts and their fractions, there was no significant difference in activity, but 65.7% or 73.4% inhibition rate was observed at high concentration (1,000 ppm) of the saponin hexane fraction (see Table 2). The results are shown in Hong HD ( Korean J Food Sci Technol 30: 1484-1487, 1998) negligence, 14.6 percent of the apple juice mixture in the SOD-like activity of the juice of fruits and vegetables, 26.7 percent for kale concentrate, 27.6% of kiwi juice mixture, in the case of non-juice mixture of 24.1 %, Respectively, as compared with the control. In addition, the above results indicate that the higher the content of polyphenol and flavonoid, the more similar activity of SOD (Kwon TD et al . , Kor J Phys Edu 3: 891-899, 2001) and previous studies (Azuma K et al . , LJ Agric Food Chem 47: 3963-3966, 1999; Lee YS et al . , Korean J Food Preserv 12: 75-79, 2005), it is thought that it contains many physiologically active substances including polyphenols and flavonoids in roots and stem of Sanjay tree. Therefore, it can be used for functional products.
아울러, 불포화 지방산의 장기보관에 따른 지질 과산화 억제율을 측정하는 항산화 측정방법인 3가철 활성 측정법을 이용하여 지질과산화 억제 활성을 측정한 결과, 본 발명의 산자나물 추출물 및 BuOH 분획물은 BHT 및 BHA와 비슷하게 높은 항 지질과산화 능력을 보였고, α-토코페롤에 비해 6배 정도 높은 지질과산화 억제 효과를 나타냈다(도 1 참조).As a result of measuring lipid peroxidation inhibitory activity using the trivalent activity measurement method, an antioxidative measurement method for measuring the inhibition rate of lipid peroxidation by long-term storage of unsaturated fatty acids, the present invention extracts and the BuOH fractions were similar to BHT and BHA Showed high antitumor lipid peroxidation ability and showed a lipid peroxidation inhibitory effect 6 times higher than that of? -Tocopherol (see FIG. 1).
상기와 같이 본 발명의 산자나무 추출물 또는 이의 분획물은 강한 전자공여능, SOD 유사 활성 및 α-토코페롤에 비해 6배 정도 높은 지질과산화 억제 효과를 나타내므로 생체 내에서 유해한 라디칼을 효과적으로 제거할 수 있는 항산화제로 사용가능하다.As described above, the present invention extracts or fractions thereof exhibit a strong electron donating ability, a SOD-like activity, and a lipid peroxidation inhibitory effect which is about 6 times higher than that of? -Tocopherol, and thus can be effectively used as an antioxidant capable of removing harmful radicals in vivo Available.
상기 항산화제는 통상적으로 사용되는 부형제, 붕해제, 감미제, 활택제, 향미제 등을 추가로 포함할 수 있으며, 통상적인 방법에 의해 정제, 캅셀제, 산제, 과립제, 현탁제, 유제, 시럽제, 기타 액제로 제형화될 수 있다. 구체적으로 상기 조성물은 경구 투여용 제형, 예를 들면 정체, 트로치제(troches), 로젠지(lozenge), 수용성 또는 우성현탁액, 조제분말 또는 과립, 에멀젼, 하드 또는 소프트 캡슐, 시럽 또는 엘릭시르제(elixirs)로 제제화된다. 정제 및 캡슐 등의 제형으로 제제하기 위해 락토오스, 사카로오스, 솔비톨, 만니톨, 전분, 아밀로펙틴, 셀롤로오스 또는 젤라틴과 같은 결합제, 디칼슘 포스페이트와 같은 부형제, 옥수수 전분 또는 고구마 전분과 같은 붕해제, 스테아르산 마그네슘, 스테아르산 칼슘, 스테아릴푸마르산 나트륨 또는 폴리에틸렌글리콜 왁스와 같은 윤활유가 함유된다. 캡슐제형의 경우는 상기에서 언급한 물질 이외에도 지방유와 같은 액체 담체를 함유한다. 또한, 상기 조성물은 경구 또는 비경구 투여할 수 있으며, 비경구 투여시 피하주사, 정맥주사, 근육내 주사 또는 흉부내 주사 주입방식을 선택하는 것이 바람직하다. 비경구 투여용 제형으로 제제화하기 위해서는 본 발명의 산자나무 추출물 또는 이의 분획물을 안정제 또는 완충제와 함께 물에서 혼합하여 현탁액으로 제조하고 이를 앰플 또는 바이알의 단위 투여형으로 제제한다. 본 발명에 따른 유효성분의 투여량은 체내에서 활성성분의 흡수도, 불활성화율 및 배설속도, 환자의 연령, 성별 및 상태, 치료할 질병의 중증 정도에 따라 적절히 선택되나, 경구 투여제의 경우 일반적으로 성인에게 1일에 체중 1 kg당 본 발명의 산자나무 추출물 또는 이의 분획물을 0.1 ~ 0.2 g의 양으로 1회 내지 수회 나누어 투여할 수 있으며, 0.1 ~ 10 ㎎의 양으로 투여하는 것이 바람직하다.
The antioxidant may further contain commonly used excipients, disintegrants, sweeteners, lubricants, flavors and the like, and may be formulated into tablets, capsules, powders, granules, suspensions, emulsions, syrups and the like by a conventional method May be formulated as a solution. Specifically, the composition may be in a form suitable for oral administration, for example, as tablets, troches, lozenges, aqueous or aqueous suspensions, pharmaceutical powders or granules, emulsions, hard or soft capsules, syrups or elixirs ). Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, celluloses or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, and disintegrants such as stearic acid Magnesium stearate, calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax. In the case of a capsule formulation, in addition to the above-mentioned substances, a liquid carrier such as fatty oil is contained. In addition, the composition may be administered orally or parenterally, and it is preferable to select subcutaneous injection, intravenous injection, intramuscular injection, or intra-thoracic injection injection method for parenteral administration. In order to formulate the formulation for parenteral administration, the extract of the present invention or its fraction is mixed with water in a stabilizer or a buffer to prepare a suspension, which is then formulated into a unit dosage form of an ampule or vial. The dosage of the active ingredient according to the present invention is appropriately selected depending on the degree of absorption, inactivation rate and excretion rate of the active ingredient in the body, the age, sex and condition of the patient, and severity of the disease to be treated. The adult of the present invention can be administered to the adult in an amount of 0.1 to 0.2 g per one kg body weight per day, and the amount of the extract is preferably 0.1 to 10 mg per day.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 활성산소가 과량으로 축적되어 발병되는 질환의 예방 및 치료용 약학적 조성물을 제공한다.
The present invention also provides a pharmaceutical composition for preventing and treating diseases in which excessive amounts of active oxygen containing an extract of the Acanthopanthis mothera or its fractions as an active ingredient are accumulated and developed.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 함유하는 노화방지용 약학적 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing aging which contains the above-described extract of Liliaceae or its fractions as an active ingredient.
본 발명의 산자나무 추출물 또는 이의 분획물은 항산화 활성을 보임으로 효과적인 활성산소가 과량으로 축적되어 발병되는 질환의 예방 및 치료용 약학적 조성물 또는 노화방지용 약학적 조성물로 유용하게 사용할 수 있다.The extracts of Wakamatsu or its fractions of the present invention exhibit antioxidative activity and thus can be effectively used as a pharmaceutical composition for preventing or treating diseases in which effective oxygen is accumulated in an excessive amount, or a pharmaceutical composition for prevention of aging.
이때, 상기 활성산소가 과량으로 축적되어 발병되는 질환은 간장 질환, 뇌졸중, 심근경색, 당뇨병성 혈관장애, 고지혈증, 급성염증, 류마티스, 암으로 이루어진 군으로부터 선택되는 것을 특징으로 한다.At this time, the disease in which the active oxygen accumulates and is caused by excessive accumulation is selected from the group consisting of liver disease, stroke, myocardial infarction, diabetic vascular disorder, hyperlipidemia, acute inflammation, rheumatism and cancer.
상기 조성물은 활성산소가 과량으로 축적되어 발병하는 질환의 예방 및 치료뿐만 아니라 노화방지를 위하여 단독으로, 또는 수술, 호르몬 치료, 약물 치료 및 생물학적 반응 조절제를 사용하는 방법들과 병용하여 사용할 수 있다.
The composition may be used alone or in combination with methods using surgery, hormone therapy, drug therapy, and biological response modifiers, as well as prevention and treatment of diseases caused by accumulation of excessive amounts of active oxygen, for prevention of aging.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 포함하는 피부 노화 방지용 화장품을 제공한다.In addition, the present invention provides a cosmetic for preventing skin aging comprising the above-mentioned extract of Liliaceae or its fractions.
상기 피부 노화 방지용 화장품으로는 로션, 연고, 겔, 크림, 패치 또는 분무제 등이 있으나 여기에 국한되는 것은 아니다. 본 발명의 상기 산자나무 추출물 또는 이의 분획물을 함유하는 피부 노화 방지용 화장품을 제조함에 있어서, 통상적으로 함유되는 피부 외용제 조성물에 본 발명의 산자나무 추출물 또는 이의 분획물이 1 내지 15 중량부, 바람직하게는 2 또는 10 중량부로 첨가할 수 있다. 본 발명의 피부용 외형제에는 본 발명의 산자나무 추출물 또는 이의 분획물에 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온 봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 피부용 외용제에 통상적으로 사용되는 임의의 다른 성분과 같은 피부 과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 또한 상기 성분들은 피부 과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다.
The skin anti-aging cosmetic may include, but is not limited to, lotions, ointments, gels, creams, patches or sprays. In producing the anti-aging skin cosmetic composition containing the above-described saponin extract of the present invention or fractions thereof, 1 to 15 parts by weight, preferably 2 to 15 parts by weight of the extract of the present invention, Or 10 parts by weight. The external composition for skin of the present invention may further contain a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, , Surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, lipophilic or lipophilic active agents, And any other ingredients conventionally used in external preparations. The components can also be introduced in amounts commonly used in the field of dermatology.
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항생제를 제공한다.In addition, the present invention provides an antibiotic comprising the above extract of Sanjobo or fractions thereof as an active ingredient.
상기 항생제는 병원성 박테리아 및 병원성 진균에 대한 항균 및 항진균 활성을 나타내는 것을 특징으로 한다. 상기 병원성 박테리아는 바실러스 서브틸러스(Bacillus subtilis), 스타필로코커스 아우레우스(Staphylococcus aureus), 대장균(Escherichia coli) 및 살모넬라 타이피무리움(Salmonella typhimurium)으로 이루어진 군으로부터 선택되는 어느 하나일 것이다. 상기 진균은 피키아 자디니(Pichia jadinii) 및 캔디다 알비칸스(Candida albicans) 중의 어느 하나일 것이다.The antibiotic is characterized by exhibiting antibacterial and antifungal activity against pathogenic bacteria and pathogenic fungi. The pathogenic bacteria are Bacillus subtilis subtilis ), Staphylococcus Staphylococcus aureus , Escherichia E. coli , and Salmonella typhimurium . The fungus is called < RTI ID = 0.0 > Pichia & jadinii ) and Candida albicans ( Candida albicans ).
본 발명의 실시예에서는 Gram(+)균인 바실러스 서브틸러스(Bacillus subtilis) 및 스타필로코커스 아우레우스(Staphylococcus aureus), Gram(-)인 대장균(Escherichia coli), 살모넬라 타이피무리움(Salmonella typhimurium), 클레브시엘라 네우모니아(Klebsiella pneumonia), 및 진균인 피키아 자디니(Pichia jadinii) 및 캔디다 알비칸스(Candida albicans)를 사용하여, 상기 산자나무 추출물 또는 이의 분획물이 항균 및 항진균제로 유용하게 사용될 수 있음을 확인하였다. 상기 바실러스 서브틸러스는 고초균으로 식품의 부패에 영향을 주고, 스타필로코커스 아우레우스는 식중독 원인균이고, 대장균은 식품오염의 지표균이며, 살모넬라 티피무리움은 식중독 미생물이고, 클레브시엘라 네우모니아는 세균성 폐렴을 일으키는 것으로 알려져 있다. 피키아 자디니는 김치에서 분리된 효모 중 하나로 알려져 있으며, 사람의 생식기나 입주위에 캔디다증(candidasis)을 일으키는 캔디다 알비칸스는 진균류(즉, 곰팡이류)에 속하는 균으로 정상적인 피부·점막·분변·객담·요 등에 존재할 수 있는 균이다. 그 결과, 클레브시엘라 네우모니아를 제외한 균들에서 전체적으로 활성을 생육 억제 활성을 나타냈다(표 3 참조). 특히 산자나무 에틸아세테이트 분획물은 대부분의 균에서 생육 억제 활성을 나타냈고, 대장균에서는 모든 산자나무 추출물 및 이의 분획물이 ≤500 ㎍/㎖에서 활성을 나타냈다(표 3 참조). 그러나 대조군인 (+)-카테킨은 본 발명의 산자나무 추출물 및 이의 분획물에 비해 항균 활성이 현저히 낮은 것으로 나타났다(표 3 참조).
In an embodiment of the present invention, Gram (+) gyunin Bacillus sub-blocks bus (Bacillus subtilis) and Staphylococcus Staphylococcus aureus ), Gram (-) Escherichia coli coli ) , Salmonella typhimurium ( Salmonella typhimurium , & lt ; RTI ID = 0.0 & gt ; Klebsiella < / RTI & gt ; pneumonia , and fungi Pichia jadinii and Candida albicans , it was confirmed that the above-mentioned extracts or fractions thereof can be effectively used as antibacterial and antifungal agents. The Bacillus subtilis is a Bacillus subtilis, which affects food spoilage. Staphylococcus aureus is a cause of food poisoning, Escherichia coli is an indicator of food contamination, Salmonella typhimurium is a food poisoning microorganism, Nia is known to cause bacterial pneumonia. Pichia jadini is known as one of the yeasts isolated from kimchi. Candida albicans, which causes candidiasis on human genital or inhabited areas, is a fungus belonging to fungi (ie, fungi) and has normal skin, mucous membrane, feces, sputum, It is a germ that can exist in urine. As a result, the activity was inhibited as a whole in the bacterium except Cleve Siella moononia (see Table 3). In particular, the ethyl acetate fraction of Sanjia wood showed the growth inhibitory activity in most of the bacterium. In E. coli, all the extracts of Sanjay tree and its fractions were active at ≤ 500 ㎍ / ㎖ (see Table 3). However, the control group (+) - catechin showed significantly lower antimicrobial activity than the potato extract of the present invention and its fractions (see Table 3).
또한, 본 발명은 상기 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 천연방부제를 제공한다.In addition, the present invention provides a natural preservative comprising the above-described extract of Sanjay or fractions thereof as an active ingredient.
본 발명의 천연방부제는 다양한 미생물, 특히 세균 및 진균에 대해 광범위한 우수한 항균 활성을 나타내었다(표 3 참조). 본 발명의 천연방부제는 상기 산자나무 추출물 또는 이의 분획물 이외에도 이의 방부 활성을 저해하지 않는 한 추가의 성분들을 포함할 수 있다. 상기한 바와 같이 광범위한 방부 스펙트럼을 갖는 본 발명에 따른 천연방부제는 의약품, 화장품, 식품, 섬유, 생활용품 등에서 방부 목적을 달성하기 위해 광범위하게 사용될 수 있으며, 이들 제품에 사용되어 제품의 품질의 안전성과 안정성을 높인다. 상기한 제품에 본 발명의 천연방부제를 포함시키는 경우, 이의 사용량은 제품의 총 중량을 기준으로 하여 0.001 내지 30 중량%, 바람직하게는 0.001 내지 20 중량 %, 더욱 바람직하게는 0.001 내지 5 중량%의 범위이다. 본 발명의 천연방부제를 포함하는 제품을 제조하기 위해, 당업자에게 널리 공지된 바에 따라 적합한 제형 및 첨가제를 선택하여 제조할 수 있을 것이다.
The natural preservative of the present invention exhibited a wide range of excellent antimicrobial activity against a variety of microorganisms, particularly bacteria and fungi (see Table 3). The natural preservatives of the present invention may contain, in addition to the above-mentioned extracts of the Acanthopanax senticosus or its fractions, additional components as long as they do not inhibit their preservative activity. As described above, the natural preservative according to the present invention having a broad spectrum of preservation can be widely used for the purpose of preservation in medicines, cosmetics, foods, fibers, household goods, etc., Increase stability. When the natural preservative of the present invention is contained in the above-mentioned products, the amount of the natural preservative is 0.001 to 30% by weight, preferably 0.001 to 20% by weight, more preferably 0.001 to 5% by weight Range. To prepare a product comprising the natural preservative of the present invention, one may make suitable formulations and additives as is well known to those skilled in the art.
또한, 본 발명은 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 항당뇨용 조성물을 제공한다.In addition, the present invention provides a composition for anti-diabetic comprising an extract of Ornithiasis or its fraction as an active ingredient.
상기 항당뇨용 조성물은 α-글루코시다아제 활성을 억제함으로써 항당뇨 활성을 나타내는 것을 특징으로 한다. 상기 α-글루코시다아제는 식이 후 섭취된 탄수화물을 소화 흡수되기 위한 상태인 단당류로 가수분해하는 역할을 갖는 장내 소화 효소로서, 상기 효소 활성을 억제하여 글루코오스 흡수를 늦춤으로써 식후 혈당 상승을 조절하는 방법이 당뇨병 치료에 적용되고 있다. 이에 본 발명의 실시예에서는 본 발명의 산자나무 추출물 또는 이의 분획물에 대한 α-글루코시다아제 저해활성을 측정함으로써 항당뇨 활성을 측정하였다. 그 결과, 산자나무 뿌리 및 줄기 BuOH 분획물에서 대조군인 당뇨치료제인 보글리보스 70.4%에 비해 각각 79.5%, 80.6%의 높은 억제율을 나타냈으나, 또 다른 대조군인 당뇨치료제인 아카보스 89%에 비해 낮은 억제율을 나타냈다(도 2 참조). 상기 결과는 본 발명의 산자나무 추출물 또는 이의 분획물이 항당뇨용 조성물로 사용될 수 있음을 나타낸다.
The antidiabetic composition is characterized by exhibiting an anti-diabetic activity by inhibiting? -Glucosidase activity. The α-glucosidase is an intestinal digestive enzyme having a role of hydrolyzing a carbohydrate taken after dieting into a monosaccharide to be digested and absorbed. The α-glucosidase is a method for controlling postprandial glucose uptake by retarding the glucose uptake by inhibiting the enzyme activity Has been applied to the treatment of diabetes. Thus, in the examples of the present invention, the antidiabetic activity was measured by measuring the inhibitory activity of? -Glucosidase on the extracts of the plants of the present invention or fractions thereof. As a result, the inhibition rates of 79.5% and 80.6% were higher than those of 70.4% of the control group, voglibose, which is the control group, in the saphenous root and stem BuOH fractions. However, the inhibition rate was lower than that of the control group, (See Fig. 2). The above results indicate that the extract of Ornithiasis or its fraction can be used as an antidiabetic composition.
아울러, 본 발명은 산자나무 추출물 또는 이의 분획물을 유효성분으로 포함하는 당뇨병 예방 및 개선용 건강식품을 제공한다.In addition, the present invention provides a health food for prevention and improvement of diabetes mellitus comprising an extract of Ornithiasis or its fraction as an active ingredient.
본 발명의 산자나무 추출물 또는 이의 분획물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 사용 목적(예방, 건강 또는 위생)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 상기 산자나무 추출물 또는 이의 분획물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양으로 첨가된다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.The extract of the present invention or fractions thereof of the present invention can be directly added or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or hygiene). Generally, in the production of a food or a drink, the said extract of the bitter gourd or fraction thereof is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material. However, in the case of long-term ingestion intended for health and hygiene purposes or health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount in the above range.
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 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 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.
본 발명의 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 0.01~0.04 g, 바람직하게는 약 0.02~0.03 g이다.The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 상기 산자나무 추출물 또는 이의 분획물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 산자나무 추출물 또는 이의 분획 또는 이의 유도체는 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01~0.1 중량부의 범위에서 선택되는 것이 일반적이다.
In addition to the above-mentioned effects, the present invention also relates to a method for preparing the above-described extracts or fractions thereof, which comprises the step of administering to the human body an effective amount of at least one compound selected from the group consisting of various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, Preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the acid extract of the present invention or a fraction thereof or a derivative thereof may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
본 발명의 산자나무 추출물 또는 이의 분획물은 항산화 효과를 나타냄으로써 항산화 또는 노화방지 화장품 또는 약학적 조성물로 유용하게 이용될 수 있고, 항균 및 항진균 효과를 나타냄으로써 항생제 또는 건강식품으로 유용하게 이용될 수 있으며, 항당뇨 효과를 나타냄으로써 항당뇨용 조성물 또는 건강식품으로 유용하게 이용될 수 있다.
INDUSTRIAL APPLICABILITY The extract of Wakamatsu or its fractions of the present invention exhibits an antioxidative effect and thus can be effectively used as an antioxidant or antioxidant cosmetic or pharmaceutical composition and exhibits antimicrobial and antifungal effects, thus being useful as an antibiotic or a health food. , And exhibit an antidiabetic effect, so that it can be usefully used as a composition for antidiabetic use or as a health food.
도 1은 3가철 활성 측정법에 의해 측정한 산자나무 추출물 및 이의 분획물의 항산화 효과를 나타낸 도이다:
a: 뿌리; 및,
b: 줄기.
도 2는 산자나무 추출물 및 이의 분획물의 α-글루코시다아제 활성을 나타낸 도이다:
a: 뿌리; 및,
b: 줄기.Figure 1 shows the antioxidant effect of the extracts of Sanjoboya and its fractions measured by trivalent active assay method:
a: root; And
b: Stem.
Fig. 2 is a diagram showing the activity of? -Glucosidase in extracts of Wakamatsu and its fractions:
a: root; And
b: Stem.
이하, 본 발명을 실시예, 실험예 및 제조예에 의해 상세히 설명한다.Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples.
단, 하기 실시예, 실험예 및 제조예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.
However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples.
<< 실시예Example 1> 시료, 기기 및 시약 1> Samples, instruments and reagents
<1-1> <1-1> 산자나무Sanjay 시료의 준비 Preparation of sample
(주)삼성생약으로부터 제공받은 산자나무(Hippophae rhamnoides L.; 비타민나무) 뿌리 및 줄기를 물로 깨끗이 세척한 후, 그늘 및 실온에서 7일간 건조하여 분쇄하여 사용하였다.( Hippophae ) supplied from Samsung Heavy Industries Co., Ltd. rhamnoides L .; Vitamin tree) roots and stems were cleaned with water, dried in shade and at room temperature for 7 days, and pulverized.
<1-2> 기기 및 시약<1-2> Devices and reagents
시약은 일급 및 특급을 사용하였고, 흡광도 측정에 사용한 UV/VIS Spectra는 Jasco사(일본) V530 분광광도계(spectrophotometer)를 사용하여 측정하였고, 회전식 감압농축기(rotary vacuum evaporator)는 EYELA사(일본)의 NE-2001과 AC-1112A를 사용하였다.The UV / VIS Spectra used for the absorbance measurement was measured using a Jasco (Japan) V530 spectrophotometer, and a rotary vacuum evaporator was measured using a spectrophotometer of EYELA (Japan) NE-2001 and AC-1112A were used.
생리활성에 사용한 DPPH(1,1-diphenyl-2-picrylhydrazyl), 파이로갈롤(Pyrogallol), Tween 20, 리놀렌산(linoleic acid), 티오시안산 암모늄(ammonium thiocyanate)을 사용하였고, 양성 대조군으로 사용한 BHA(tert-butyl hydroxyanisole), BHT(tert-butyl hydroxytoluene), α-토코페롤(α-tocopherol), 아스코르브산(Ascorbic acid), (+)-카테킨(catechin), 케토코나졸(Ketoconazol), 마이코스탄틴(Mycostantin), 테트라시클린(tetracycline)은 Sigma사(USA) 제품을 사용하였다.
DPPH (1,1-diphenyl-2-picrylhydrazyl), Pyrogallol,
<< 실시예Example 2> 2> 산자나무Sanjay 추출물 및 이의 Extract and its 분획물의Fraction 제조 Produce
<2-1> 메탄올 추출물 제조<2-1> Preparation of methanol extract
실시예 1-1의 방법으로 준비한 산자나무 뿌리(408 g) 및 줄기(115 g) 가루를 100% 메탄올 10배에 침지하여 실온에서 7일간 3번 반복 추출하고 여과지에 여과하였다. 상기 추출액을 용액이 완전히 증발할 때까지 감압 하에 농축기를 사용하여 농축함으로써, 건조된 추출물을 수득하였다. 상기 방법을 이용한 결과 산자나무 뿌리(408 g) 및 줄기(115 g) 가루에서 추출물 뿌리 68 g 및 줄기 60 g을 제조할 수 있었다.(408 g) and stem (115 g) prepared by the method of Example 1-1 were immersed in 100% methanol 10 times, repeatedly extracted three times at room temperature for 7 days, and filtered through a filter paper. The extract was concentrated using a concentrator under reduced pressure until the solution completely evaporated to obtain a dried extract. As a result of using the above method, 68 g of extract root and 60 g of stem were able to be prepared from the seeds of 408 g of the Japanese peony root and 115 g of the stem.
<2-2> 물 추출물 제조<2-2> Preparation of water extract
추출용매로 물을 사용하여, 상기 실시예 <2-1>의 추출방법과 동일하게 추출하여 추출물 7.3 및 1.8 g을 제조할 수 있었다.Using water as an extraction solvent, extraction was carried out in the same manner as in Example <2-1> to obtain 7.3 and 1.8 g of extracts.
<2-3> 에탄올 추출물 제조<2-3> Preparation of ethanol extract
추출용매로 100% 에탄올을 사용하여, 상기 실시예 <2-1>의 추출방법과 동일하게 추출하여 추출물 9.2 및 2.5 g을 제조할 수 있었다.The extraction was carried out in the same manner as in Example <2-1> above using 100% ethanol as an extraction solvent, and extracts 9.2 and 2.5 g were obtained.
<2-4> <2-4> 분획물Fraction 제조 Produce
실시예 2-1의 방법으로 수득한 추출물을 증류수에 현탁시킨 후 n-헥산(hexane), 에틸 아세테이트(ethyl acetate), 부탄올(butanol) 순으로 계통 분획하였다. 3회 반복한 후, 각 용매 분획물을 감압 농축하였다.The extract obtained in Example 2-1 was suspended in distilled water and fractionated in the order of n -hexane, ethyl acetate and butanol. After repeating 3 times, each solvent fraction was concentrated under reduced pressure.
그 결과, 각각 n-헥산[뿌리(3.9 g), 줄기(5.1 g)], 에틸 아세테이트[뿌리(2 g), 줄기(4.4 g)], 부탄올[뿌리(14 g), 줄기(11.1 g)]의 용매 분획물을 수득하였다. 상기 방법을 이용한 결과 산자나무 뿌리(408 g) 및 줄기(115 g) 가루에서 추출물 뿌리 68 g 및 줄기 60 g을 제조할 수 있었다.
As a result, n - hexane [root (3.9 g), stem (5.1 g)], ethyl acetate [root (2 g), stem (4.4 g)], butanol [root (14 g) ] ≪ / RTI > As a result of using the above method, 68 g of extract root and 60 g of stem were able to be prepared from the seeds of 408 g of the Japanese peony root and 115 g of the stem.
<< 실험예Experimental Example 1> 1> 전자공여능Electron donating ability 조사 Research
Xiong Q 등(Biological and Pharmaceutical Bulletin 19:1580-1585, 1996)에 의해 제시된 DPPH 자유 라디칼 소거법에 의한 전자공여능 측정방법을 이용하였다.Xiong Q, etc. ( Biological and Pharmaceutical Bulletin 19: 1580-1585, 1996) was used to determine the electron donating ability by the DPPH free radical scavenging method.
구체적으로, 4 ㎖의 메탄올이 담긴 시험관에 실시예 2의 방법으로 수득한 실험군으로서 산자나무 뿌리 및 줄기 추출물 및 이의 분획물, 양성 대조군(BHA, BHT, α-토코페롤) 및 음성 대조군(무첨가군)을 농도별로 각각 첨가하여 시료를 준비하였다. 상기 시료에 DPPH(0.15 mM) 용액 1 ㎖을 첨가한 후, 실온에서 30분간 방치하여 반응시키고 517 ㎚에서 UV/VIS 분광광도계로 흡광도를 측정하였다. 이때, 상기 시료가 DPPH 자유 라디칼을 50% 억제하는데 필요한 시료의 농도(RC50; ㎍/㎖)는 음성 대조군의 DPPH 자유 라디칼 값을 50% 감소시키는데 필요한 시료의 농도로 나타내었다.Specifically, as the experimental group obtained by the method of Example 2, 4 ml of methanol was added to the test tube, and the roots and the stem extracts and fractions thereof, the positive control (BHA, BHT, alpha-tocopherol) and the negative control group Respectively, to prepare samples. 1 ml of a solution of DPPH (0.15 mM) was added to the sample, the reaction was allowed to stand at room temperature for 30 minutes, and the absorbance was measured with a UV / VIS spectrophotometer at 517 nm. At this time, the concentration of the sample (RC 50 ; / / ml) required for the sample to inhibit the DPPH free radical by 50% was expressed as the concentration of the sample required to reduce the DPPH free radical value of the negative control by 50%.
실험군의 항산화 활성을 기존의 항산화제인 BHA, BHT, α-토코페롤과 비교한 결과, 표 1에 나타난 바와 같이 모든 산자나무 뿌리 및 줄기의 추출물 및 이의 분획물에서 높은 전자공여능을 나타냈으며, 특히 뿌리 및 줄기의 EtOAc(에틸 아세테이트) 분획에서 각각 RC50이 1.5 ± 0.5 ㎍/㎖ 및 1.0 ± 0.5 ㎍/㎖로 이는 대조군 α-토코페롤 RC50 12 ± 0.5 ㎍/㎖과 비교해 볼 때 더 강한 전자공여능을 나타냈다.As shown in Table 1, the antioxidative activity of the experimental group was higher than that of the conventional antioxidants BHA, BHT and α-tocopherol, and thus showed high electron donating ability in all the extracts of roots and stems and their fractions, RC 50 was 1.5 ± 0.5 ㎍ / ㎖ and 1.0 ± 0.5 ㎍ / ㎖, respectively, in the EtOAc (ethyl acetate) fraction, which showed stronger electron donating ability compared to the control α-tocopherol RC 50 12 ± 0.5 ㎍ / ㎖.
1)RC50 : 30분 후 DPPH의 50% 감소에 필요한 양, 각 수치들은 세 번 반복된 검사에 의해 평균 ± 표준편차로 나타났다.
1) RC 50 : the amount required for a 50% reduction in DPPH after 30 minutes, each value was expressed as mean ± SD by three repeated tests.
<< 실험예Experimental Example 2> 활성산소제거효소( 2> Reactive oxygen scavenging enzyme ( SuperoxideSuperoxide dismutasedismutase ; ; SODSOD ) 유사 활성 조사) Similar activity studies
Marklund 등(Marklund S & Marklund G, Eur J Biochem 47:468-474, 1975)의 방법에 따라 SOD 유사 활성을 측정하였다.Marklund et al. (Marklund S & Marklund G, Eur J Biochem 47: 468-474, 1975).
구체적으로, 실시예 2의 방법으로 수득한 실험군으로써 농도별(100, 1,000, 5,000 및 10,000 ppm) 산자나무 뿌리 및 줄기 추출물 및 이의 분획물 및 대조군(무첨가군) 각각 0.2 ㎖에 Tris-HCl의 완충용액(50 mM Tris + 10 mM EDTA, pH 8.5) 2.6 ㎖이 혼합된 시료에 7.2 mM 파이로갈롤 0.2 ㎖을 가하였다. 25℃에서 10분간 방치함으로써 반응시킨 후, 1.0 N HCl 0.1 ㎖를 가하여 반응을 정지시켰고 반응액 중 산화된 파이로갈롤의 양을 420 ㎚에서 측정하였다. SOD 유사 활성은 하기 수학식 1에 따라 실험군과 대조군의 흡광도 차이를 백분율(%)로 나타낸 값이다.Specifically, as a test group obtained by the method of Example 2, 0.2 ml of each of the concentrations of (100, 1,000, 5,000 and 10,000 ppm) waxy root and stem extract and its fractions and the control group (no added group) And 2.6 ml of a buffer solution (50 mM Tris + 10 mM EDTA, pH 8.5), 0.2 ml of 7.2 mM pyrogallol was added. The reaction was allowed to stand at 25 DEG C for 10 minutes, after which 0.1 mL of 1.0 N HCl was added to stop the reaction, and the amount of oxidized pyrogallol was measured at 420 nm in the reaction solution. The SOD-like activity is a value indicating the difference in absorbance between the experimental group and the control group as a percentage (%) according to the following equation (1).
그 결과, 표 2에 나타난 바와 같이 산자나무에서 메탄올 추출물을 비롯한 모든 분획물에서 농도가 증가할수록 활성산소제거효소(Superoxide dismutase; SOD) 유사 활성이 농도 의존적으로 증가하였다. 산자나무 뿌리 및 줄기 추출물 및 이의 분획물은 각각 저농도(100 ppm)에서는 활성이 큰 차이를 보이지 않았으나, 산자나무 뿌리 또는 줄기 헥산 분획물 1,000 ppm에서는 각각 65.7% 또는 73.4% 억제율을 나타났다.As a result, as shown in Table 2, the concentration of the superoxide dismutase (SOD) -like activity increased in a concentration-dependent manner in all the fractions including methanol extracts from Sanjay trees. In the low concentration (100 ppm), no significant difference was observed in the activity of the extracts from the root and stem extracts and the fractions thereof, respectively, whereas the inhibition rate was 65.7% or 73.4% at 1,000 ppm of the roots or stem hexane fractions, respectively.
(ppm)density
(ppm)
1) 모든 수치들은 세 번 반복된 검사에 의해 평균 ± 표준편차로 나타났다.
1) All values were expressed as mean ± standard deviation by three repeated tests.
<< 실험예Experimental Example 3> 지질과산화 억제 활성 조사 3> Lipid peroxidation inhibition activity
Inatani R 등(Agricultural and Biological Chemistry 47:521-528, 1983)에 의한 3가철 활성 측정법을 이용하여 지질과산화 억제 활성을 측정하였다. 상기 3가철 활성 측정법은 불포화 지방산의 장기보관에 따른 지질 과산화 억제율을 측정하는 항산화 측정방법이다.Inatani R, etc. (Agricultural and Biological Chemistry 47: 521-528, 1983) was used to measure lipid peroxidation inhibitory activity. The trivalent activity measurement method is an antioxidant measuring method for measuring the inhibition rate of lipid peroxidation by long-term storage of unsaturated fatty acids.
구체적으로, 불포화 지방산인 리놀렌산(2.51%), 에탄올(2.0 ㎖), 0.05 M 인산염 완충용액(pH 7.0, 4.0 ㎖), H2O(1.9 ㎖) 및 10% 트윈 20(0.1 ㎖)이 담긴 20 ㎖ 시험관에, 전량이 10 ㎖이 되고 산자나무 뿌리 및 줄기 추출물 및 이의 분획물, 양성 대조군(BHA, BHT, α-토코페롤) 및 대조군(무첨가군)을 각각 최종농도가 0.005%가 되도록 넣은 후, 40℃의 암소 배치하였다. 상기 혼합액 시료 0.1 ㎖에 75% 에탄올(9.7 ㎖) 및 30% 티오시아산 암모늄(0.1 ㎖)을 가하였고, 2 × 10-2 M 염화제일철의 3.5% 염산용액(0.1 ㎖)을 가한 후, 정확히 3분 후에 500 ㎚에서 UV/VIS 분광광도계로 흡광도를 측정하였다. 이어서, 48시간마다 총 35일간 측정하였다. 지질 과산화물 생성 억제능(%)은 하기 수학식 2에 따라 계산하였다.Specifically, a solution containing 20% (w / v) of linolenic acid (2.51%), unsaturated fatty acid, ethanol (2.0 ml), 0.05 M phosphate buffer solution (pH 7.0, 4.0 ml), H 2 O (1.9 ml) (BHA, BHT,? -Tocopherol) and a control group (no-added group) were added to a final concentration of 0.005%, and the resulting solution was added to 40 ml of a solution of 40 ≪ / RTI > 75% ethanol (9.7 ml) and 30% ammonium thiocyanate (0.1 ml) were added to 0.1 ml of the mixed solution sample. To the mixture was added 3.5 ml of a hydrochloric acid solution (0.1 ml) of 2x10 -2 M ferrous chloride After 3 minutes, the absorbance was measured with a UV / VIS spectrophotometer at 500 nm. Then, a total of 35 days was measured every 48 hours. The inhibitory activity (%) of lipid peroxide formation was calculated according to the following equation (2).
그 결과, 도 1에 나타난 바와 같이 α-토코페롤은 17일 이후 급격한 산화가 이뤄져 BHT 및 BHA 비해 항 지질과산화 능력이 낮게 나타났다. 산자나무 뿌리(도 1a)에서는 BuOH 분획물 및 MeOH 추출물이 양성 대조군인 BHT 및 BHA와 유사한 높은 항 지질과산화 능력을 나타냈고, 줄기(도 1b) 또한 BuOH 분획물 및 MeOH 추출물에서 양성 대조군인 BHT 및 BHA와 비슷하게 높은 항 지질과산화 능력을 보였다. 또한, BuOH 분획물과 MeOH 추출물이 줄기 및 뿌리에서 α-토코페롤에 비해 6배 정도 높은 지질과산화 억제 효과를 나타냈다.
As a result, as shown in FIG. 1,? -Tocopherol was abruptly oxidized after 17 days, indicating that the lipid peroxidation capacity was lower than that of BHT and BHA. BuOH fraction and MeOH extract showed high antitumor lipid peroxidation ability similar to that of positive control groups BHT and BHA, and stem (FIG. 1B) also showed positive control groups BHT and BHA in the BuOH fraction and MeOH extract Showed similarly high lipid peroxidation capacity. In addition, BuOH fraction and MeOH extract showed 6 times higher inhibition effect on lipid peroxidation than α - tocopherol in stem and root.
<< 실험예Experimental Example 4> 항균활성 조사 4> Antimicrobial activity investigation
Kobayasi A 등(Zeitschrift fur Naturforsch 51:527-533, 1996)의 연속 2배 희석법(serial 2-fold dilution)에 따라 항균활성을 측정하였다.Kobayasia et al. ( Zeitschrift fur Naturforsch 51: 527-533, 1996) in a serial 2-fold dilution.
<4-1> <4-1> 피검체Subject
Gram(+)균인 바실러스 서브틸러스(Bacillus subtilis) 및 스타필로코커스 아우레우스(Staphylococcus aureus), Gram(-)인 대장균(Escherichia coli), 살모넬라 타이피무리움(Salmonella typhimurium), 클레브시엘라 네우모니아(Klebsiella pneumonia), 및 진균인 피키아 자디니(Pichia jadinii) 및 캔디다 알비칸스(Candida albicans)를 사용하였다.The Gram (+) strain Bacillus subtilis subtilis ) and Staphylococcus Staphylococcus aureus ), Gram (-) Escherichia coli coli ) , Salmonella typhimurium ( Salmonella typhimurium , Klebsiella pneumonia , and fungus Pichia < RTI ID = 0.0 > jadinii ) and Candida albicans were used.
<4-2> 배양 및 활성 측정<4-2> Culture and activity measurement
포도상구균(micrococcus)은 영양(nutrient) YM 배지에 배양하여 상기 균체 현탁액을 직경 25 ㎜ 시험관에 10 ㎖씩 접종하여 각각 37℃와 30℃에서 12시간 동안 진탕 배양(100 rpm)하였다. 상기 배양액을 각각의 배지에 100배 희석하여 항균시험에 사용하였다. 시료를 96 웰 마이크로 분석 플레이트의 제 1번 구에 넣고 조제된 균체 현탁액을 분주하여 2배씩 희석하여 사용하였다.Staphylococcus micrococcus was cultured in nutrient YM medium, and 10 ml of the cell suspension was inoculated into a 25 mm diameter test tube and shake cultured (100 rpm) at 37 ° C and 30 ° C for 12 hours, respectively. The culture broth was diluted 100 times in each medium and used for the antibacterial test. Samples were placed in No. 1 wells of a 96-well microanalysis plate and the prepared cell suspension was dispensed and diluted 2-fold.
상기의 시료를 37℃와 30℃에서 24시간 암 조건에서 배양하여 세균의 증식을 육안으로 탁도 정도를 측정함으로써 확인하였다. 항균 활성은 세균의 생육을 억제하는 최저농도(Minimum Inhibitory Concentration: 이하, MIC)를 측정하였다(Kim MJ & Hyun JO, Kor J Vreed 29:115-123, 1997).The above samples were incubated at 37 ° C and 30 ° C for 24 hours under dark conditions, and the proliferation of bacteria was visually confirmed by measuring the degree of turbidity. The minimum inhibitory concentration (MIC) that inhibited the growth of bacteria was measured (Kim MJ & Hyun JO, Kor J Vreed 29: 115-123,1997).
그 결과, 표 3에 나타난 바와 같이 클레브시엘라 네우모니아를 제외한 균들에서 전체적으로 생육 억제 활성을 나타냈다. 특히 산자나무 뿌리 및 줄기의 에틸아세테이트 분획물은 대부분의 균에서 생육 억제 활성을 나타냈다. 그중 대장균에서는 모든 산자나무 추출물 및 이의 분획물이 ≤500 ㎍/㎖에서 활성을 나타냈다. 대조군인 (+)-카테킨은 본 발명의 산자나무 추출물 및 이의 분획물에 비해 항균 활성이 현저히 낮은 것으로 나타났다.As a result, as shown in Table 3, the growth inhibitory activity was shown as a whole in the bacterium except Cleve C. laneumonia. Especially, the ethyl acetate fraction of the roots and stems of the wild plants showed the growth inhibitory activity in most bacteria. Among them, all of the extracts and their fractions were active at ≤ 500 ㎍ / ㎖ in E. coli. The control group, (+) - catechin, was significantly lower in antimicrobial activity than the extract of the present invention and its fractions.
1) B.s.: Bacillus subtilis , S.a.: Staphylococus aureus , E.c.: Escherichia coli, S.t.: Salmonella typhimurium, K.p.: Klebsiella pneumonia, P.j.: Pichia jadinii 및 C.a.: Candida albicans.
1) Bs: Bacillus subtilis , Sa: Staphylococus aureus , Ec: Escherichia coli, St: Salmonella typhimurium, Kp: Klebsiella pneumonia, Pj: Pichia jadinii and Ca: Candida albicans .
<< 실험예Experimental Example 5> 5> 항당뇨Anti-diabetic 활성 조사 Activity investigation
α-글루코시다아제는 식이 후 섭취된 탄수화물을 소화 흡수되기 위한 상태인 단당류로 가수분해하는 역할을 갖는 장내 소화 효소로서 글루코오스 흡수를 늦춤으로써 식후 혈당 상승을 조절하는 방법이 당뇨병 치료에 적용되고 있다. 상기 α-글루코시다아제 저해활성을 측정함으로써 항당뇨 활성을 측정하였다.α-glucosidase is a intestinal digestive enzyme that hydrolyzes carbohydrates taken after dieting into a monosaccharide, which is a state for digestion and absorption. As a result, a method of controlling postprandial increase in blood glucose by delaying glucose absorption has been applied to the treatment of diabetes. The anti-diabetic activity was measured by measuring the? -Glucosidase inhibitory activity.
실시예 2의 방법으로 수득한 산자나무 뿌리 및 줄기 추출물을 1 ㎎/㎖의 농도가 되도록 0.3 M KPB 완충용액(pH 7.0)으로 희석한 후, α-글루코시다아제 효소액(Sigma Chemical, USA) 100 ㎕를 넣고 혼합하여 37℃에서 30분간 반응시켰다. 상기 반응액을 100℃에서 5분간 두어 반응을 정지시킨 후 상기 반응액 40 ㎕에 글루코오스 키트(Lot.R658, 아산제약(주)) 200 ㎕를 첨가하여 37℃에서 5분간 발색 시킨 후 496 ㎚에서 흡광도를 측정하였다. 이때, 산자나무 추출물 및 이의 분획물의 흡광도에 대한 양성 대조군으로 당뇨치료제인 아카보스(Acarbose) 및 보글리보스(Voglibose)를 사용하였다. 상기 아카보스는 α-글루코시다아제 억제제로 시판되고 있는 의약품으로서 주로 소장 세포의 점막에 존재하는 말타아제(maltase)의 활성을 억제하는 것으로 알려져있다(Carrascosa JM et al ., Diabetes Obes Metab 3: 240-248, 2001). α-글루코시다아제에 대한 저해활성(억제율)을 수학식 3에 따라 계산하였다.The roots and stem extracts obtained by the method of Example 2 were diluted with 0.3 M KPB buffer solution (pH 7.0) to a concentration of 1 mg / ml, and then treated with? -Glucosidase enzyme solution (Sigma Chemical, USA) 100 Mu] l, and the mixture was reacted at 37 [deg.] C for 30 minutes. The reaction solution was kept at 100 ° C for 5 minutes to stop the reaction. 200 μl of a glucose kit (Lot.R658, ASAN PHARMACEUTICAL CO., LTD.) Was added to 40 μl of the reaction solution, followed by color development at 37 ° C for 5 minutes. Absorbance was measured. At this time, acarbose and Voglibose, which are diabetic therapeutic agents, were used as a positive control for the absorbance of the extract of Sanjay tree and its fractions. It is known that acarbose inhibits the activity of maltase existing in the mucosa of small intestinal cells as a commercially available drug as an? -Glucosidase inhibitor (Carrascosa JM et al . , Diabetes Obes Metab 3: 240-248, 2001). The inhibitory activity (inhibition rate) against? -glucosidase was calculated according to Equation (3).
그 결과, 도 2에 나타난 바와 같이 산자나무 뿌리 및 줄기 BuOH 분획물에서 대조군 보글리보스 70.4%에 비해 각각 79.5%, 80.6%의 높은 억제율을 나타냈으나, 아카보스 89%에 비해 낮은 억제율을 나타냈다.
As a result, as shown in Fig. 2, the inhibition rates of 79.5% and 80.6% were higher than those of the control group, 70.4%, respectively, but the inhibition rate was lower than that of acarbose 89%.
<< 제제예Formulation example 1> 약학적 제제의 제조 1> Preparation of pharmaceutical preparations
1. 산제의 제조1. Manufacturing of powder
산자나무 추출물 또는 분획물 2 g2 grams of Lycopersiconnet extract or fraction
유당 1 gLactose 1 g
상기의 성분을 혼합하고 기밀포에 충진하여 산제를 제조하였다.
The above components were mixed and packed in airtight bags to prepare powders.
2. 정제의 제조2. Preparation of tablets
산자나무 추출물 또는 분획물 100 ㎎100 mg of the extract or fraction
옥수수전분 100 ㎎
유 당 100 ㎎100 mg of milk
스테아린산 마그네슘 2 ㎎2 mg of magnesium stearate
상기의 성분을 혼합한 후, 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.
After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.
3. 캡슐제의 제조3. Preparation of capsules
산자나무 추출물 또는 분획물 100 ㎎100 mg of the extract or fraction
옥수수전분 100 ㎎
유 당 100 ㎎100 mg of milk
스테아린산 마그네슘 2 ㎎2 mg of magnesium stearate
상기의 성분을 혼합한 후, 통상의 캡슐제의 제조방법에 따라서 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.
After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.
4. 환의 제조4. Manufacture of rings
산자나무 추출물 또는 분획물 1 g1 g of Lycopersiconnet extract or fraction
유당 1.5 gLactose 1.5 g
글리세린 1 gGlycerin 1 g
자일리톨 0.5 g0.5 g of xylitol
상기의 성분을 혼합한 후, 통상의 방법에 따라 1 환 당 4 g이 되도록 제조하였다.
After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.
5. 과립의 제조5. Manufacture of granules
산자나무 추출물 또는 분획물 150 ㎎150 mg of the extract or fraction
대두 추출물 50 ㎎Soybean extract 50 mg
포도당 200 ㎎200 mg of glucose
전분 600 ㎎600 mg of starch
상기의 성분을 혼합한 후, 30% 에탄올 100 ㎎을 첨가하여 섭씨 60℃에서 건조하여 과립을 형성한 후 포에 충진하였다.
After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.
<< 제제예Formulation example 2> 2> 화장료의Cosmetic 제조 Produce
<2-1> 유연 화장수의 제조<2-1> Production of Flexible Lotion
산자나무 추출물 또는 분획물을 유효성분으로 함유하는 유연 화장수의 제제예는 다음 표 4와 같이 제조하였다.Examples of the preparation of a soft lotion containing an extract of Sanjayaku or fractions as an active ingredient were prepared as shown in Table 4 below.
<2-2> 영양 크림의 제조<2-2> Preparation of nutritional cream
산자나무 추출물 또는 분획물을 함유한 영양크림의 제제예는 다음 표 5의 조성과 같이 제조하였다.Formulation examples of nutritional creams containing extracts or fractions were prepared according to the composition shown in Table 5 below.
<제제예 3> 식품의 제조 ≪ Formulation Example 3 > Preparation of food
본 발명의 산자나무 추출물 또는 분획물을 포함하는 식품들을 다음과 같이 제조하였다.Foods comprising the extracts or fractions of the present invention were prepared as follows.
1. 조리용 양념의 제조1. Preparation of cooking seasoning
본 발명의 산자나무 추출물 또는 분획물 20~95 중량부로 건강 증진용 조리용 양념을 제조하였다.Healthy cooking sauces were prepared with 20 to 95 parts by weight of the extracts or fractions of the present invention.
2. 밀가루 식품의 제조2. Manufacture of flour food
본 발명의 산자나무 추출물 또는 분획물 0.5~5.0 중량부를 밀가루에 첨가하고, 이 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하여 건강 증진용 식품을 제조하였다.0.5 to 5.0 parts by weight of the extract of Orangan tree of the present invention was added to wheat flour, and bread, cake, cookies, crackers and noodles were prepared by using this mixture to prepare a food for health promotion.
3. 유제품(dairy products)의 제조3. Manufacture of dairy products
본 발명의 산자나무 추출물 또는 분획물 5~10 중량부를 우유에 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.5 to 10 parts by weight of the extract or fraction of the present invention was added to milk and various dairy products such as butter and ice cream were prepared using the milk.
4. 선식의 제조4. Manufacture of wire
현미, 보리, 찹쌀, 율무를 공지의 방법으로 알파화시켜 건조한 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.Brown rice, barley, glutinous rice, and yulmu were alfalted by a known method and dried, and the powder was prepared into a powder having a particle size of 60 mesh by a pulverizer.
검정콩, 검정깨, 들깨도 공지의 방법으로 쪄서 건조한 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.Black soybeans, black sesame seeds, and perilla seeds were steamed and dried in a known manner, and were then ground to a powder having a particle size of 60 mesh.
상기에서 제조한 곡물류, 종실류 및 본 발명의 산자나무 추출물 또는 분획물의 건조분말을 다음의 비율로 배합하여 제조하였다.The grains, the seeds, and the dry powder of the extracts or fractions of the present invention were mixed in the following proportions.
곡물류(현미 30 중량부, 율무 15 중량부, 보리 20 중량부),(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)
종실류(들깨 7 중량부, 검정콩 8 중량부, 검정깨 7 중량부),Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)
산자나무 추출물 또는 분획물(3 중량부),(3 parts by weight), < RTI ID = 0.0 >
영지(0.5 중량부),(0.5 part by weight),
지황(0.5 중량부)
(0.5 parts by weight)
<< 제제예Formulation example 4> 음료의 제조 4> Manufacturing of beverages
1. 건강음료의 제조1. Manufacture of health drinks
산자나무 추출물 또는 분획물 1000 ㎎ 1000 mg of Lycopersiconazole extract or fraction
구연산 1000 ㎎ Citric acid 1000 mg
올리고당 100 g 100 g of oligosaccharide
매실농축액 2 g Plum concentrate 2 g
타우린 1 g Taurine 1 g
정제수를 가하여 전체 900 ㎖ Purified water was added to a total of 900 ml
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 ℓ용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다. The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, It is used in the production of the health beverage composition of the invention.
상기 조성비는 비교적 기호 음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용 용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.
Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.
Claims (10)
An antioxidant composition comprising, as an active ingredient, an ethyl acetate fraction obtained by dissolving a roots or a stem methanol extract in water followed by sequential use of n-hexane, ethyl acetate and butanol.
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