KR101680294B1 - Method for producing ginseng extract - Google Patents
Method for producing ginseng extract Download PDFInfo
- Publication number
- KR101680294B1 KR101680294B1 KR1020140056937A KR20140056937A KR101680294B1 KR 101680294 B1 KR101680294 B1 KR 101680294B1 KR 1020140056937 A KR1020140056937 A KR 1020140056937A KR 20140056937 A KR20140056937 A KR 20140056937A KR 101680294 B1 KR101680294 B1 KR 101680294B1
- Authority
- KR
- South Korea
- Prior art keywords
- ginseng
- ginseng extract
- ginsenoside
- ginsenosides
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- Prior art date
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- 235000020710 ginseng extract Nutrition 0.000 title claims abstract description 52
- 238000004519 manufacturing process Methods 0.000 title abstract description 17
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- 108090000790 Enzymes Proteins 0.000 claims abstract description 30
- 235000003140 Panax quinquefolius Nutrition 0.000 claims description 38
- 235000008434 ginseng Nutrition 0.000 claims description 38
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 claims description 37
- 239000000203 mixture Substances 0.000 claims description 29
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- 238000000034 method Methods 0.000 claims description 14
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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 인삼 추출물의 제조방법에 관한 것이다.
상기와 같은 본 발명에 따르면 다당류 가수분해 효소(PHE, Polysaccharide Hydrolyzing Enzymes)를 이용함으로써, 간단한 방법으로 고함량의 다양한 진세노사이드를 추출함과 동시에 오염발생을 줄이고 비용을 절감하는 효과가 있다.
또한, 본 발명은 고함량의 메이저 진세노사이드뿐만 아니라 마이너 진세노사이드도 함유되어 Rg1 및 Rb1의 비율과 Re, Rc, Rb2, Rg2, Rf, Rd, Rh1 및 화합물 k(compound K)의 함량이 높은 상품성이 우수한 인삼 추출물 및 상기 인삼 추출물이 함유된 건강기능식품, 피부 외용제 조성물 및 약학 조성물을 제공하는 효과가 있다. The present invention relates to a method for producing ginseng extract.
According to the present invention, by using polysaccharide hydrolyzing enzymes (PHE), various ginsenosides can be extracted in a simple manner, and contamination occurrence and cost can be reduced.
The present invention also relates to a pharmaceutical composition containing not only a high amount of major ginsenoside but also minor ginsenosides, wherein the ratio of Rg1 and Rb1 and the content of Re, Rc, Rb2, Rg2, Rf, Rd, Rh1 and compound K A ginseng extract excellent in high commerciality and a health functional food, an external preparation for skin and a pharmaceutical composition containing the ginseng extract are provided.
Description
본 발명은 인삼 추출물의 제조방법에 관한 것으로서, 더욱 상세하게는 세포벽 분해효소(PHE)를 이용하여 진세노사이드가 다량 함유된 인삼 추출물을 제조하는 방법에 관한 것이다.The present invention relates to a method for producing ginseng extract, and more particularly, to a method for producing a ginseng extract containing a large amount of ginsenoside using cell wall degrading enzyme (PHE).
인삼으로부터 추출된 추출물은 면역, 항암, 항산화, 항당뇨 활성을 포함한 여러 가지 의학적인 효과가 있는 것으로 알려져 있는데, 이러한 효과는 주로 인삼 사포닌인 진세노사이드라 불리우는 트리테르페노이드 글루코사이드(triterpenoid glucoside)에 기인한다. 진세노사이드는 식물에 존재하는 여러 사포닌 중 인삼의 사포닌 성분만을 특별하게 구분하여 명명한 것으로써, 진세노사이드는 다양한 항암, 항알레르기, 항염증, 정신안정, 진통, 기억력 개선, 간기능 강화, 항당뇨, 항스트레스, 호흡기 질환 개선, 소화기관 개선, 혈당 강하, 면역력 증강, 항산화 및 항노화 등의 효과가 있는 것으로 보고되어 있다.The extracts extracted from ginseng are known to have various medical effects including immunity, anticancer, antioxidant and antidiabetic activity. This effect is mainly caused by triterpenoid glucoside, which is called ginseng saponin, do. Ginsenoside is a special kind of saponin which is found in various saponins in plants. Ginsenoside has various anti-cancer, antiallergic, anti-inflammatory, mental stability, analgesic, memory, Anti-diabetic, anti-stress, improvement of respiratory diseases, digestive system improvement, blood glucose lowering, immunity enhancement, antioxidant and anti-aging.
진세노사이드는 1960년대 초반에 분리되어 생리학적, 약리학적 연구에 널리 이용되었으며, 다양한 생물학적 활성을 갖는 40 개 이상의 진세노사이드가 인삼 뿌리에서 분리되었다. 진세노사이드는 아글리콘(aglycon)의 구조에 따라 크게 프로토파낙사디올(glucosides of protopanaxadiol, PPD-type ginsenosides, Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, Rg3), 프로토파낙사트리올(glucosides of protopanaxatriol, PPT-type ginsenosides, Re, Rf, Rgl, Rg2) 및 올레아난(Oleanane-type ginsenosides) 세 가지로 나뉜다.Ginsenosides were isolated in the early 1960s and widely used in physiological and pharmacological studies. More than 40 ginsenosides with various biological activities were isolated from ginseng roots. The ginsenosides were classified as aglycon according to the structure of the aglycon as glucosides of protopanaxadiol (PPD-type ginsenosides, Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd and Rg3) There are three types of glycosides: protopanaxatriol, PPT-type ginsenosides, Re, Rf, Rgl, Rg2 and oleanane-type ginsenosides.
또한, 진세노사이드는 비율상 많은 양을 차지하고 있는 프로토파낙사트리올(PPT) 계열의 Rg1, Re 등과 프로토파낙사디올(PPD) 계열의 Rb1, Rc, Rb2, Rd 등으로 이루어진 메이저 진세노사이드(major ginsenoside)와 Rg3, Rk, Rg5 등 미량의 마이너 진세노사이드(minor ginsenoside)로 나뉜다. 마이너 진세노사이드는 메이저 진세노사이드의 당 가수분해 처리공정에 의해 만들어지기도 하며, 자연계에 존재하는 배당체 화합물과 마찬가지로 메이저 진세노사이드에 비하여 흡수나 약효 등의 면에서 훨씬 뛰어난 효과를 나타낸다. In addition, ginsenoside is a major ginsenoside consisting of Rg1 and Re of protopanaxyl triol (PPT) and Rb1, Rc, Rb2 and Rd of protopanaxadiol (PPD) (major ginsenoside) and minor ginsenosides such as Rg3, Rk, and Rg5. Minor ginsenosides may be produced by the sugar hydrolysis process of major ginsenosides and, as with the glycoside compounds present in nature, exhibit far superior effects in terms of absorption and efficacy compared to major ginsenosides.
프로토파낙사디올계 진세노사이드인 Rb1, Rb2, Rc, Rd는 사람의 장내 세균에 의해 20-O-β-D-글루코피라노실(glucopyranosyl)-20(S)-프로토파낙사디올(prototopanaxadiol)로 대사되고(Hasegawa H, et al., Planta Medica 63:463-40, 1997; Tawab MA, et al., Drug Metab. Dispos. 31:1065-71, 2003), 프로토파낙사트리올계 진세노사이드인 Re, Rg1은 장내세균에 의해 진세노사이드 Rh1이나 진세노사이드 F1으로 대사되며(Hasegawa H, et al., Planta Medica 62:453-7, 1996; Tawab MA, et al., Drug Metab. Dispos. 31:1065-71, 2003), 이렇게 전환된 화합물 K, Rh1 및 F1은 다양한 생리활성을 나타낸다. Rb1, Rb2, Rc, and Rd, protopanaxadiol-based ginsenosides, were identified as 20-O- beta -D-glucopyranosyl-20 (S) -prototopanaxadiol by human intestinal bacteria. (Hasegawa H, et al., Planta Medica 63: 463-40, 1997; Tawab MA, et al., Drug Metab. Dispos. 31: 1065-71, 2003), protopanaxatriol ginsenosides Re, Rg1 is metabolized by intestinal bacteria to ginsenoside Rh1 or ginsenoside F1 (Hasegawa H, et al., Planta Medica 62: 453-7, 1996; Tawab MA, et al., Drug Metab. Dispos 31: 1065-71, 2003). The compounds K, Rh1 and F1 thus converted exhibit various physiological activities.
종래 알려진 화합물 K를 제조하는 방법으로는 다이올계 사포닌을 효소인 나린지나제로 가수분해시켜 제조하거나, 쥐에 경구투여 후 대장에서 분해산물로 생성되는 화합물 K를 분리하는 방법이 있다. 그러나, 이 방법들은 수율이 극히 낮을 뿐만 아니라 여러 가지 2차 대사산물이 생성되어 화합물 K를 고순도로 정제하기가 어려운 문제점이 있으며(Kohda Hiroshi and Tanaka Osamu, The Pharm. Society of Japan 95:246(1975); Karikura 등, Chem. Pharm. Bull. 38:2859(1990)), 인삼 사포닌을 화합물 k로 전환하는 방법은 주로 미생물에 의해 이루어져 혐기성 조건에서 균을 배양해야 하거나 식용이 어려운 문제점이 있다. A known method for producing the compound K is a method in which a diol-based saponin is prepared by enzymatic digestion with NARIN or by hydrolysis, or a compound K produced as a degradation product in the large intestine after oral administration to a mouse. However, these methods have a problem that not only the yield is extremely low but also various secondary metabolites are generated, making it difficult to purify the compound K with high purity (Kohda Hiroshi and Tanaka Osamu, The Pharm. Society of Japan 95: 246 ), Karimura et al., Chem. Pharm. Bull. 38: 2859 (1990)), the method of converting ginseng saponin into compound k is mainly performed by microorganisms.
상기와 같은 생리학적, 약리학적 유효성분인 진세노사이드가 함유된 인삼 추출물을 제조하기 위해 다양한 방법이 개발되었고, 이는 진세노사이드의 추출 효율을 최대화하고 분해를 최소화하며 유기용매 사용으로 인한 환경오염과 생산비용을 줄이는 방안으로 개발되어 왔다. 인삼 추출물에 대한 관심과 니즈(needs)가 증가하고 있고 인삼 추출물의 효능 강화를 위해서는 흡수가 용이한 진세노사이드를 고함량으로 추출해야 한다. 따라서, 우수한 인삼 제품을 제조하기 위해서는 메이저 진세노사이드뿐만 아니라 수삼과 같은 인삼에는 함유되어 있지 않거나 미량으로 함유되어 다양한 약리 작용을 하는 것으로 알려져 있는 마이너 진세노사이드도 별도의 전환과정 없이 함께 추출 및 제조할 수 있는 저비용, 고효율의 방법을 개발할 필요가 있다.Various methods have been developed for producing ginsenosides containing the physiological and pharmacological active ingredient ginsenosides as described above. These methods maximize the extraction efficiency of ginsenosides, minimize the decomposition and minimize the environmental pollution And to reduce production costs. Interest and needs for ginseng extracts are increasing, and in order to enhance the efficacy of ginseng extract, ginsenosides that are easy to absorb should be extracted in high amounts. Therefore, in order to produce excellent ginseng products, minor ginsenoside, which is known to have various pharmacological actions not contained in ginseng such as ginseng, as well as major ginsenosides, There is a need to develop a low-cost, high-efficiency method that can be used.
선행기술로 대한민국 등록특허 10-0228510(진세노사이드 Rg3 및/또는 Rg5의 제조방법), 대한민국 공개특허 10-2013-0059773(피부 주름 개선효과를 가지는 가공 인삼 추출물을 유효성분으로 함유하는 건강기능성식품), 대한민국 공개특허 10-2011-0108820(이산화탄소 초임계 추출된 인삼 추출물을 포함하는 비만 억제용 조성물) 등이 있으나, 종래 용매를 이용한 가열 추출방법은 시간이 오래 소요되고 효율이 떨어질 뿐만 아니라 유효성분이 열에 의해 파괴되는 문제점이 있다. 또한, 초음파를 활용한 추출방법은 음파 진동의 기계적 충격에 의해 세포막이 파괴되어 내용물의 용출은 용이하게 하나 산업적으로 적용하기에는 어려운 문제가 있고, 이산화탄소를 이용한 초임계 추출방법은 지용성 물질 추출에는 적정하나 수용성 물질 추출에는 한계가 있다. 그 밖에 효소를 이용한 진세노사이드 제조방법에 관한 관련 선행기술로 한국 공개특허 제10-2013-0057536호(로하멘트 CL을 이용한 진세노사이드 컴파운드 케이의 제조방법), 한국 등록특허 제10-0318304호(진세노사이드알지1의제조방법) 등이 있다.Korean Patent No. 10-0228510 (method for producing ginsenosides Rg3 and / or Rg5) and Korean Patent Laid-Open No. 10-2013-0059773 (a health functional food containing the processed ginseng extract having the effect of improving skin wrinkles as an active ingredient) ), Korean Patent Publication No. 10-2011-0108820 (a composition for suppressing obesity containing supercritical carbon dioxide supercritical extract), and the like. However, the conventional heating and extraction method using a solvent takes a long time and low efficiency, There is a problem that it is destroyed by heat. In addition, the extraction method using ultrasonic waves has a problem that the cell membrane is broken due to the mechanical impact of the sound wave vibration and the dissolution of the contents is easy, but it is difficult to apply industrially. Supercritical extraction method using carbon dioxide is suitable for extracting lipophilic substances Extraction of water-soluble materials is limited. Other related arts related to the production method of ginsenosides using enzymes include Korean Patent Laid-Open No. 10-2013-0057536 (production method of ginsenoside compound k using Lohament CL), Korean Patent No. 10-0318304 (Production method of Ginsenoside algin 1), and the like.
본 발명의 목적은 다당류 가수분해 효소(PHE, Polysaccharide Hydrolyzing Enzymes)를 이용하여 간단하게 저비용으로 진세노사이드 함량이 높은 인삼 추출물을 제조하는 방법을 제공함에 있다.It is an object of the present invention to provide a method for producing a ginseng extract having a high ginsenoside content at low cost simply by using polysaccharide hydrolyzing enzymes (PHE).
또한, 본 발명의 목적은 고함량의 메이저 진세노사이드가 함유된 인삼 추출물뿐만 아니라 마이너 진세노사이드가 다량 함유된 인삼 추출물을 제공하며, 구체적으로 Rg1 및 Rb1의 비율과 Re, Rc, Rb2, Rg2, Rf, Rd, Rh1, 화합물 k(compound K)의 함량이 높은 상품성이 우수한 인삼 추출물 및 상기 인삼 추출물이 함유된 건강기능식품, 피부 외용제 조성물 및 약학 조성물을 제공함에 있다.It is another object of the present invention to provide ginseng extract having a high content of major ginsenoside as well as ginseng extract containing a large amount of minor ginsenoside. Specifically, the ratio of Rg1 and Rb1 and the ratio of Re, Rc, Rb2, Rg2 , Rf, Rd, Rh1 and compound k (compound K), and a health functional food, an external preparation for skin and a pharmaceutical composition containing the ginseng extract.
상기 목적을 달성하기 위하여, 본 발명은 (1) 인삼속(Panax species) 식물의 뿌리, 줄기, 잎 및 열매를 포함하는 군에서 선택되는 1 이상의 원료를 용매와 혼합하여 현탁액을 제조하는 단계; (2) 상기 현탁액에 셀룰라아제(cellulase), 아밀라아제(amylase), 펙티나아제(pectinase), 셀로비아제(cellobiase) 및 비스코자임(viscozyme)을 포함하는 군에서 선택되는 1 이상의 당 분해효소를 첨가하여 반응시키는 단계; 및 (3) 상기 반응 종료 후 효소를 불활성화시키는 단계;를 포함하는 인삼 추출물의 제조방법을 제공한다.(1) preparing a suspension by mixing at least one raw material selected from the group consisting of root, stem, leaf and fruit of a plant of Panax species with a solvent; (2) adding at least one saccharide-degrading enzyme selected from the group consisting of cellulase, amylase, pectinase, cellobiase and viscozyme to the suspension, ; And (3) deactivating the enzyme after the reaction is completed.
상기 (1)단계에서 용매는 물 및 탄소수 1 내지 4의 저급 알코올을 포함하는 군에서 선택된 1 이상인 것을 특징으로 한다.In the step (1), the solvent is at least one selected from the group consisting of water and lower alcohols having 1 to 4 carbon atoms.
상기 (1)단계에서 현탁액은 용매 100 중량부에 대하여 인삼속 식물의 뿌리, 줄기, 잎 및 열매를 포함하는 군에서 선택되는 1 이상의 원료를 10 내지 25 중량부로 혼합한 것을 특징으로 한다.In the step (1), the suspension is mixed with 10 to 25 parts by weight of at least one raw material selected from the group consisting of root, stem, leaf and fruit of a plant of the genus Ginseng, relative to 100 parts by weight of the solvent.
상기 (2)단계에서 당 분해효소는 현탁액 100 부피부에 대하여 1 내지 5 부피비로 혼합하는 것을 특징으로 한다.In the step (2), the saccharide-degrading enzyme is mixed with 1 to 5 parts by volume of the skin of 100 parts of the suspension.
상기 (2)단계에서 반응 시간 및 온도는 5 내지 80 시간 및 20 내지 60 ℃인 것을 특징으로 한다.In the step (2), the reaction time and temperature are 5 to 80 hours and 20 to 60 ° C.
상기 (2)단계에서 당 분해효소로 아밀라아제와 펙티나아제를 첨가하여 진세노사이드 Rg1 및 Rb1이 다량 함유된 인삼 추출물을 제조하는 것을 특징으로 한다.In step (2), amylase and pectinase are added as glucosease to produce ginseng extract containing ginsenosides Rg1 and Rb1 in a large amount.
이때, 제조된 진세노사이드 Rg1 및 Rb1이 다량 함유된 인삼 추출물은 기능성 음료 조성물로 이용하는 것을 특징으로 한다.At this time, the ginseng extract containing a large amount of the ginsenosides Rg1 and Rb1 is used as a functional beverage composition.
상기 (2)단계에서 당 분해효소로 셀룰라아제를 첨가하여 진세노사이드 Rd가 다량 함유된 인삼 추출물을 제조하는 것을 특징으로 한다.In step (2), a ginseng extract containing a large amount of ginsenoside Rd is prepared by adding a cellulase as a saccharide-degrading enzyme.
이때, 제조된 진세노사이드 Rd가 다량 함유된 인삼 추출물은 피부 외용제 조성물로 이용하는 것을 특징으로 한다.At this time, the ginseng extract containing a large amount of the produced ginsenoside Rd is used as a composition for external application for skin.
상기 (2)단계에서 당 분해효소로 비스코자임, 셀룰라아제 및 셀로비아제를 첨가하여 진세노사이드 Rh1이 다량 함유된 인삼 추출물을 제조하는 것을 특징으로 한다.In step (2), biscozyme, cellulase, and cellulase are added as a saccharide decomposing enzyme to produce a ginseng extract containing a large amount of ginsenoside Rh1.
이때, 제조된 진세노사이드 Rh1이 다량 함유된 인삼 추출물은 항암용 약학 조성물로 이용하는 것을 특징으로 한다.At this time, the ginseng extract containing a large amount of ginsenoside Rh1 is used as an anticancer pharmaceutical composition.
상기 (2)단계에서 당 분해효소로 셀룰라아제, 아밀라아제 및 셀로비아제를 첨가하여 화합물 K(compound K)가 다량 함유된 인삼 추출물을 제조하는 것을 특징으로 한다.In step (2), a ginseng extract containing a large amount of the compound K is prepared by adding cellulase, amylase and cellobiase as a glucosease.
이때, 제조된 화합물 K가 다량 함유된 인삼 추출물은 항암용 약학 조성물로 이용하는 것을 특징으로 한다.At this time, the ginseng extract containing a large amount of the prepared compound K is used as an anticancer pharmaceutical composition.
상기 (3)단계에서 효소를 불활성화시키기 위한 온도 및 시간은 70 내지 100 ℃ 및 10 내지 50 분인 것을 특징으로 한다.The temperature and time for inactivating the enzyme in the step (3) are 70 to 100 ° C and 10 to 50 minutes.
상기와 같은 본 발명에 따르면 다당류 가수분해 효소(PHE, Polysaccharide Hydrolyzing Enzymes)를 이용함으로써, 간단한 방법으로 고함량의 다양한 진세노사이드를 추출함과 동시에 오염발생을 줄이고 비용을 절감하는 효과가 있다.According to the present invention, by using polysaccharide hydrolyzing enzymes (PHE), various ginsenosides can be extracted in a simple manner, and contamination occurrence and cost can be reduced.
또한, 본 발명은 고함량의 메이저 진세노사이드가 함유된 인삼추출물뿐만 아니라 마이너 진세노사이드가 다량 함유된 인삼추출물을 제공하며, 구체적으로 Rg1 및 Rb1의 비율과 Re, Rc, Rb2, Rg2, Rf, Rd, Rh1, 화합물 k(compound K)의 함량이 높은 상품성이 우수한 인삼 추출물 및 상기 인삼 추출물이 함유된 건강기능식품, 피부 외용제 조성물 및 약학 조성물을 제공하는 효과가 있다. In addition, the present invention provides a ginseng extract having a high content of major ginsenosides as well as a ginseng extract containing a large amount of minor ginsenosides. Specifically, the ratio of Rg1 and Rb1 and the ratio of Re, Rc, Rb2, Rg2, Rf , Rd, Rh1, and compound k (compound K), and a health functional food, a skin external composition and a pharmaceutical composition containing the ginseng extract.
도 1은 (a) 효소 미처리된 인삼 추출물, (b) 아밀라아제 + 펙티나아제 처리된 인삼 추출물, (c) 표준 진세노사이드 혼합물에 함유된 진세노사이드를 나타내는 크로마토그램.
도 2는 다양한 효소의 조합에 따른 진세노사이드 Rg1 및 Rb1의 함량 비교.
도 3은 셀룰라아제 처리시 진세노사이드 Rb1의 Rd로의 전환을 나타내는 크로마토그램.
도 4는 셀룰라아제 처리시 진세노사이드 함량 비교.
도 5는 비스코자임, 셀룰라아제 및 셀로비아제를 18 및 24 시간 동안 처리시 진세노사이드 Rh1이 생성됨을 나타내는 TLC 크로마토그램.
도 6은 비스코자임, 셀룰라아제 및 셀로비아제를 18 및 24 시간 동안 처리시 진세노사이드 Rh1이 생성됨을 나타내는 진세노사이드 Rh1 함량 비교.
도 7은 인삼 추출물의 제조 공정도.Fig. 1 is a chromatogram showing (a) ginsenoside not contained in enzyme-treated ginseng extract, (b) amylase + pectinase-treated ginseng extract, and (c) standard ginsenoside mixture.
Figure 2 compares the content of ginsenosides Rg1 and Rb1 with various combinations of enzymes.
3 is a chromatogram showing the conversion of ginsenoside Rb1 to Rd during cellulase treatment.
Figure 4 compares ginsenoside content in cellulase treatment.
Figure 5 is a TLC chromatogram showing the production of ginsenoside Rh1 upon treatment of viscose, cellulase and cellobiase for 18 and 24 hours.
Figure 6 compares ginsenoside Rh1 content, which shows that ginsenoside Rh1 is produced when treating viscose, cellulase and cellulase for 18 and 24 hours.
Fig. 7 is a view showing a manufacturing process of ginseng extract. Fig.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
인삼은 약 60 cm 내외의 다년생 초본으로, 보통 4~6년 후 가을에 수확하며 가공하지 않은 그대로의 인삼을 수삼이라 한다. 수삼은 저장에 문제가 있기 때문에 가공이 필요하며 가공방식에 따라 백삼, 홍삼 등으로 분류되고, 생육환경에 따라 재배삼, 장뇌삼, 산삼 등으로 구분된다.Ginseng is a perennial herb, about 60 cm in length, usually harvested 4 to 6 years later in autumn, and raw ginseng is called raw ginseng. Ginseng is classified as white ginseng and red ginseng according to the processing method and it is classified into cultivation ginseng, ginseng ginseng, and wild ginseng depending on the growing environment.
인삼은 오래 전부터 천연 에르고제닉 에이드(natural ergogenic aid)로 알려져 있으며, 면역력 증강, 항스트레스, 항암, 항노화, 항산화, 숙취, 강장효과 등에 이르기까지 다양한 효능들이 밝혀지고 있다(Kim SH, et al., J Sports Med. Phys. Fitness., 45(2):178-82, 2005)(Gillis CN, Biochem Pharmacol., 54(1):1-8, 1997; Attele AS, et al., Biochem Pharmacol., 58(11):1685-93, 1999; Shin HR, et al., Cancer Causes Control., 11(6):565-76, 2000). PPD계 진세노사이드와 PPT계 진세노사이드는 체내에서 각기 다른 작용을 하는데, 진세노사이드 Rb1으로 대표되는 PPD계 사포닌은 중추신경에 대한 억제작용을 나타내어 정신안정, 신경이완, 진통, 항경련, 혈압강하, 파파베린양 작용 등을 나타내는 것으로 알려져 있고, 진세노사이드 Rg1으로 대표되는 PPT계 사포닌은 중추신경에 대해 흥분적으로 작용하여 항피로작용이 있는 것으로 알려져 있다.Ginseng has long been known as a natural ergogenic aid and has been shown to have various effects ranging from immunity enhancement, anti-stress, anti-cancer, anti-aging, antioxidant, hangover and tonic effects (Kim SH, et al. Biochem Pharmacol., 54 (1): 1-8, 1997; Attele AS, et al., Biochem Pharmacol., 45 (2): 178-82, 2005). , Shin et al., Cancer Causes Control, 11 (6): 565-76, 2000). PPD ginsenoside and PPT ginsenoside have different functions in the body. PPD saponin, represented by ginsenoside Rb1, has an inhibitory action on the central nervous system and is known to have mental stability, nervous relaxation, analgesia, Blood pressure lowering, and papaverine positive action. PPT saponin represented by ginsenoside Rg1 is known to have an antipyrotic action by acting excitably on the central nervous system.
미토콘드리아 에너지 대사를 개선시키고, 유산소 운동능력(aereobic exercise performance)의 증진에 진세노사이드 Rg1과 Rb1이 작용한다고 보고된 바 있다(Wang LC, et al., Planta Med., 64(2):130-133, 1998). 진세노사이드 Rg1은 면역증강효과(Okamura N et al. Biol. Pharm. Bull. 17(2), 270(1994)), 항피로작용(Saito H et al. Japan J. Pharmacology, 24, 119(1974)) 및 유해자극 방어작용(Yoshimatsu H et al. Physiology and Behavior, 53(1), 1(1993))을 갖고, 진세노사이드 Rd는 부신피질호르몬 분비 촉진작용(Hiai S et al. Chem Pharm. Bull. 31, 168(1983)), 진세노사이드 Rb1은 단백질 합성 촉진작용(Zhang J T et al. Acta Pharmaceutica Sinica 23(1), 12,(1988)), 진세노사이드 Rc는 진통작용, 단백질 및 지질 합성 촉진작용, 진세노사이드 Rb2는 항당뇨작용, 면역조절작용, 콜레스테롤 대사작용 등 다양한 약리효과를 갖는다.It has been reported that ginsenosides Rg1 and Rb1 work in improving mitochondrial energy metabolism and promoting aereobic exercise performance (Wang LC, et al., 64 (2): 130- 133, 1998). Ginsenoside Rg1 has been shown to be effective in the enhancement of immune function (Okamura N et al. Biol. Pharm. Bull. 17 (2), 270 (1994)), antipyrotic action (Saito H et al. Japan J. Pharmacology, ), And a noxious stimulant defense action (Yoshimatsu H et al. Physiology and Behavior, 53 (1), 1 (1993)) and ginsenoside Rd has a stimulating action on adrenocortical hormone secretion (Hiai S et al. Bax. 31, 168 (1983)), ginsenoside Rb1 is a protein synthesis promoting action (Zhang JT et al. Acta Pharmaceutica Sinica 23 (1), 12 Lipid synthesis promoting action, and ginsenoside Rb2 have various pharmacological effects such as antidiabetic action, immunoregulatory action, and cholesterol metabolism.
인삼 추출물의 피부와 관련된 효능으로는 소염작용(Korean J. Dermatol. 18(1):39-42(1980), Korean J. Dermatol. 14(4):335-339(1976)), 과각화방지(Korean J. Dermatol. 28(4):434-440(1990)), 여드름 예방 및 치료(Korean J. Dermatol. 30(1):27-33(1992), Korean J. Dermatol. 28(4):434-440(1990)), 항산화(Proceedings of the 2 nd international ginseng symposium 13-17(1978)), 탄력성 및 수화성 증가(Fitoterapia 57(1):15-28(1986), Fitoterapia 57(4) 217-222(1986)), 상처치유(Brit. J. Pharmacol. 125:255-262(1998), Arch. pharm. res. 25(1):71-76(2002)), 콜라겐 분해억제(Mol. Cells 9(5):476-483(1999)), 미백(Journal of the society of cosmetic scientists of korea 27(2):45-56(2001)) 등이 있다.The effects of ginseng extracts on the skin include anti-inflammatory effects (Korean J. Dermatol. 18 (1): 39-42 (1980), Korean J. Dermatol.14 (4): 335-339 (1976)), Korean J. Dermatol. 28 (4): 434-440 (1990)), acne prevention and treatment (Korean J. Dermatol.30 (1): 27-33 (1992) (Fitoterapia 57 (1): 15-28 (1986), Fitoterapia 57 (4)), antioxidant activity (Proceedings of the 2nd International Ginseng Symposium 13-17 217-222 (1986)), wound healing (Brit. J. Pharmacol. 125: 255-262 (1998), Arch Pharm Res. 25 (1): 71-76 Cells 9 (5): 476-483 (1999)) and whitening (Journal of the Society of Cosmetic Scientists of Korea 27 (2): 45-56 (2001)).
화합물 K는 종양의 침입을 막거나 염색체 변형과 종양 형성을 예방함으로써 전이 또는 항암 효과를 유도하고(Wakabayashi C, et al., Oncol. Res. 9:411-7, 1998; Lee SJ, et al., Cancer Lett. 144:39-43, 1999), Rh1은 각종 암세포 성장에 대한 세포독성 효과(Odashima S, et al., Cancer Res. 45:2781-4, 1985; Ota T, et al., Cancer Res. 47:3863-7, 1987; Lee HY, et al., Differentiation mechanism of ginsenosides in cultured murine F9 teratocarcinoma stem cells. Proc. 6th Int. Ginseng symp. Seoul 127-31, 1993) 및 항알레르기, 항염증(Park EK, et al., Int. Arch. Allergy Immunol. 133:113-120, 2004) 활성을 가진다.Res. 9: 411-7, 1998; Lee SJ, et al., ≪ RTI ID = 0.0 > 1998). ≪ / RTI > , Cancer Lett. 144: 39-43, 1999), Rh1 is a cytotoxic effect for various cancer cell growth (Odashima S, et al., Cancer Res. 45: 2781-4, 1985; Ota T, et al., Cancer Ginseng symp. Seoul 127-31, 1993) and anti-allergic, anti-inflammatory (anti-inflammatory, anti-inflammatory) (Park EK, et al., Int. Arch. Allergy Immunol., 133: 113-120, 2004).
본 발명은 (1) 인삼속(Panax species) 식물의 뿌리, 줄기, 잎 및 열매를 포함하는 군에서 선택되는 1 이상의 원료를 용매와 혼합하여 현탁액을 제조하는 단계; (2) 상기 현탁액에 셀룰라아제(cellulase), 아밀라아제(amylase), 펙티나아제(pectinase), 셀로비아제(cellobiase) 및 비스코자임(viscozyme)을 포함하는 군에서 선택되는 1 이상의 당 분해효소를 첨가하여 반응시키는 단계; 및 (3) 상기 반응 종료 후 효소를 불활성화시키는 단계;를 포함하는 인삼 추출물의 제조방법을 제공한다.(1) preparing a suspension by mixing at least one raw material selected from the group consisting of roots, stems, leaves and fruits of a plant of Panax species with a solvent; (2) adding at least one saccharide-degrading enzyme selected from the group consisting of cellulase, amylase, pectinase, cellobiase and viscozyme to the suspension, ; And (3) deactivating the enzyme after the reaction is completed.
상기 (1)단계에서 인삼속 식물로는 고려인삼(Panax ginseng), 화기삼(Panax quinquefolium), 전칠삼(Panax notoginseng), 죽절삼(Panax japonicum), 삼엽삼(Panax trifolium), 히말라야삼(Panax pseudoginseng) 또는 베트남삼(Panax vietnamensis) 등이 있고, 가공여부와 생육환경에 따른 홍삼, 수삼, 백삼, 재배삼, 장뇌삼 또는 산삼 등 품종이나 종류에 제한 없이 이용이 가능하다.Panax ginseng, Panax quinquefolium, Panax notoginseng, Panax japonicum, Panax trifolium, and Panax pseudoginseng in the step (1) Or Vietnam (Panax vietnamensis). It can be used without limitation in variety and kinds such as red ginseng, fresh ginseng, white ginseng, cultivated ginseng, camellia ginseng, or wild ginseng depending on the processing condition and growth environment.
상기 (1)단계에서 용매는 물 및 메탄올, 에탄올, 부탄올 등의 탄소수 1 내지 4의 저급 알코올을 포함하는 군에서 선택된 1 이상인 것이 바람직하다.In the step (1), the solvent is preferably at least one selected from the group consisting of water and a lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol, and butanol.
상기 (1)단계에서 현탁액은 용매 100 중량부에 대하여 인삼속 식물의 뿌리, 줄기, 잎 및 열매를 포함하는 군에서 선택되는 1 이상의 원료가 10 내지 25 중량부, 바람직하게는 20 내지 25 중량부로 혼합된 것이 최적의 효과를 나타낸다. 용매에 대해 인삼 원료의 농도가 10 내지 25 중량부일 경우 진세노사이드의 수율이 향상되어 제조공정에 있어 효율성이 높아지고 진세노사이드의 함량을 증가시켜 주는 효과가 있다. In the step (1), the suspension contains 10 to 25 parts by weight, preferably 20 to 25 parts by weight, of at least one raw material selected from the group consisting of roots, stems, leaves and fruits of the plant of the genus Ginseng Mixed shows the best effect. When the concentration of the ginseng raw material in the solvent is 10 to 25 parts by weight, the yield of ginsenoside is improved, thereby increasing the efficiency in the manufacturing process and increasing the content of ginsenoside.
상기 (1)단계에서 현택액은 생리학적으로 허용되는 산을 첨가하여 pH를 4.5 내지 5, 바람직하게는 4.5 내지 4.8로 조절하는 것이 최적의 효과를 나타낸다. pH는 효소 활성에 직접적인 영향을 미치는 중요인자로서 pH의 작은 변화에 따라 추출되는 진세노사이드의 종류와 수율이 현저하게 달라지게 된다. 본 발명에서 셀룰라아제, 아밀라아제, 펙티나아제, 비스코자임 및 셀로비아제를 포함하는 군에서 선택되는 1 이상의 당 분해효소가 인삼 추출과정에서 소량의 첨가에도 최적의 활성을 나타내는 pH의 범위는 4.5 내지 5로서, 진세노사이드 수율이 향상되는 효과가 있다.In the step (1), the pH value is adjusted to 4.5 to 5, preferably 4.5 to 4.8 by adding a physiologically acceptable acid to the present solution. pH is an important factor that directly affects the enzyme activity, and the type and yield of ginsenoside extracted depend on the small change of pH. In the present invention, at least one saccharide-degrading enzyme selected from the group consisting of cellulase, amylase, pectinase, viscose and cellobiase has a pH ranging from 4.5 to 5 , There is an effect that the yield of ginsenoside is improved.
이때, 생리학적으로 허용되는 산은 염산, 황산 및 인산을 포함하는 무기산과 옥살산, 아세트산, 숙신산, 타르타르산, 아스코르브산, 시트르산, 글루탐산, 메타설폰산 및 에탄설폰산을 포함하는 유기산으로 이루어진 군에서 선택되는 1 이상인 것이 바람직하다.Wherein the physiologically acceptable acid is selected from the group consisting of inorganic acids including hydrochloric acid, sulfuric acid and phosphoric acid and organic acids including oxalic acid, acetic acid, succinic acid, tartaric acid, ascorbic acid, citric acid, glutamic acid, methanesulfonic acid and ethanesulfonic acid 1 or more.
상기 (2)단계에서 당 분해효소는 현탁액 100 부피부에 대하여 1 내지 5 부피비, 바람직하게는 2 내지 4 부피비, 더 바람직하게는 3 내지 4 부피부로 혼합하는 것이 진세노사이드 추출에 있어서 최적의 효과를 나타낸다. 본 발명에서 당 분해효소는 인삼 추출물에 함유된 진세노사이드의 종류 및 함량에 결정적인 영향을 미치는 요인으로서, 현탁액 100 부피부에 대하여 1 내지 5 부피비로 당 분해효소를 첨가할 경우 추출하고자 하는 진세노사이드의 수율이 향상되어 제조공정에 있어 효율성이 높아지고 진세노사이드의 함량을 증가시켜 주는 효과가 있다. In step (2), the saccharide-degrading enzyme is mixed with 100 parts of the suspension in an amount of 1 to 5 parts by volume, preferably 2 to 4 parts by volume, more preferably 3 to 4 parts by weight, Effect. In the present invention, the carbohydrase has a decisive influence on the kind and content of ginsenoside contained in the ginseng extract. When the carbohydrase is added in a volume ratio of 1 to 5 per 100 parts of the suspension, The yield of the side is improved, thereby increasing the efficiency in the manufacturing process and increasing the content of ginsenoside.
상기 (2)단계에서 반응 시간 및 온도는 5 내지 80 시간, 바람직하게는 18 내지 48 시간, 더욱 바람직하게는 24 내지 36 시간 및 20 내지 60 ℃인 것이 최적의 효과를 나타낸다. 상기 반응 시간 및 온도 조건에서 당 분해효소를 처리할 경우 제조공정의 경제성, 효율성뿐만 아니라 진세노사이드 수율의 향상과 함께 목적으로 하는 진세노사이드의 추출이 가능하다.In the step (2), the reaction time and temperature are optimal for 5 to 80 hours, preferably 18 to 48 hours, more preferably 24 to 36 hours and 20 to 60 ° C. When the glycosidase is treated under the above reaction time and temperature conditions, it is possible to extract the desired ginsenoside in addition to the economical efficiency and the efficiency of the production process as well as the yield of ginsenoside.
상기 (2)단계에서 당 분해효소로 아밀라아제와 펙티나아제를 첨가하여 진세노사이드 Rg1 및 Rb1이 다량 함유된 인삼 추출물을 제조할 수 있다. In step (2), amylase and pectinase may be added as a saccharide-degrading enzyme to produce a ginseng extract containing a large amount of ginsenosides Rg1 and Rb1.
본 발명은 상기 제조된 진세노사이드 Rg1 및 Rb1이 다량 함유된 인삼 추출물에서 얻은 Rg1 및 Rb1을 유효성분으로 함유하는 기능성 음료 조성물을 제공한다. 더욱 구체적으로, 항피로, 항산화, 면역력 증진 등을 위한 기능성 음료 조성물로 이용될 수 있으며, 이는 식약처가 제시한 진세노사이드 Rg1, Rb1 및 Rg3의 함량이 2.5 내지 34 mg/g일 경우 면역력 증진, 피로 개선, 항산화 등 기능성 건강기능식품으로 인정되는 것에 부합한다.The present invention provides a functional beverage composition containing Rg1 and Rb1 obtained from a ginseng extract containing a large amount of ginsenosides Rg1 and Rb1 as an active ingredient. More specifically, it can be used as a functional beverage composition for anti-fatigue, antioxidation, immunity enhancement and the like. When the content of ginsenosides Rg1, Rb1 and Rg3 suggested by the food supplier is 2.5 to 34 mg / g, Fatigue improvement, antioxidation, and other functional health functional foods.
또한, 상기 (2)단계에서 당 분해효소로 셀룰라아제를 첨가하여 진세노사이드 Rd가 다량 함유된 인삼 추출물을 제조할 수 있다.In addition, in step (2), a ginseng extract containing a large amount of ginsenoside Rd can be prepared by adding a cellulase with a glucosease.
본 발명은 상기 제조된 진세노사이드 Rd가 다량 함유된 인삼 추출물에서 얻은 Rd을 유효성분으로 함유하는 피부 외용제 조성물을 제공한다. 구체적으로, 미백, 보습, 여드름 개선, 항산화, 노화방지, 항균, 항염, 아토피 등을 위한 피부 외용제 조성물로 이용될 수 있다(Kim et al., J Ginseng Res. (2013) 37:54-63).The present invention provides an external preparation for skin comprising Rd obtained from ginseng extract containing a large amount of ginsenoside Rd as an active ingredient. Specifically, it can be used as a composition for external application for skin for whitening, moisturizing, acne improvement, antioxidation, anti-aging, antibacterial, antiinflammatory, atopic and the like (Kim et al., J Ginseng Res. (2013) 37: 54-63) .
또한, 상기 (2)단계에서 당 분해효소로 비스코자임, 셀룰라아제 및 셀로비아제를 첨가하여 진세노사이드 Rh1이 다량 함유된 인삼 추출물을 제조할 수 있다.In addition, in step (2), ginseng extract containing ginsenoside Rh1 in a large amount can be prepared by adding biscozyme, cellulase, and cellulase as a glucosease.
본 발명은 상기 제조된 진세노사이드 Rh1이 다량 함유된 인삼 추출물에서 얻은 Rh1을 유효성분으로 함유하는 항암용 약학 조성물을 제공한다.The present invention provides an anticancer pharmaceutical composition containing Rh1 obtained from ginseng extract containing the ginsenoside Rh1 as an active ingredient.
또한, (2)단계에서 당 분해효소로 셀룰라아제, 아밀라아제 및 셀로비아제를 첨가하여 화합물 K(compound K)가 다량 함유된 인삼 추출물을 제조할 수 있다.In addition, in step (2), ginseng extract containing a large amount of compound K may be prepared by adding cellulase, amylase and cellobiase as a saccharide-degrading enzyme.
본 발명은 상기 제조된 화합물 K가 다량 함유된 인삼 추출물에서 얻은 화합물 K를 유효성분으로 함유하는 항암용 약학 조성물을 제공한다.The present invention provides an anticancer pharmaceutical composition containing, as an active ingredient, a compound K obtained from a ginseng extract containing a large amount of the compound K prepared above.
본 발명은 Rh1 또는 화합물 K가 다량 함유된 인삼 추출물을 제조함으로써, 메이저 진세노사이드의 추출과 함께 메이저 진세노사이드를 마이너 진세노사이드로 전환시켜 보다 간단한 방법으로 Rh1 및 화합물 k의 제조가 가능한 효과가 있다.The present invention relates to a method for producing Rh1 and a compound K, which comprises preparing a ginseng extract containing a large amount of Rh1 or a compound K, thereby extracting major ginsenosides and converting major ginsenosides to minor ginsenosides, .
상기 (3)단계에서 효소를 불활성화시키기 위한 온도 및 시간은 70 내지 100 ℃ 및 10 내지 50 분간, 바람직하게는 70 내지 90 ℃에서 10 내지 40 분간 열처리하는 것이 최적의 효과를 나타낸다. 상기 온도 및 시간 조건에서 효소를 불활성화시킬 경우 진세노사이드 함량의 변화를 최소화할 수 있는 효과가 있다. 또한, 효소를 불활성화시킨 다음 48 내지 72 시간 동안 동결건조하여 농축하는 것이 바람직하다.The temperature and time for inactivating the enzyme in the step (3) are optimum for effecting the heat treatment at 70 to 100 ° C for 10 to 50 minutes, preferably at 70 to 90 ° C for 10 to 40 minutes. When the enzyme is inactivated under the above temperature and time conditions, the change of the ginsenoside content can be minimized. In addition, it is preferable that the enzyme is inactivated and then lyophilized for 48 to 72 hours for concentration.
하기에 본 발명의 인삼 추출물을 포함하는 여러 가지 제제예 및 제형예를 설명하고 있으나, 이는 본 발명을 제한하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Various formulation examples and formulations including the ginseng extract of the present invention are described below, but the present invention is not intended to be limited thereto but is specifically described.
제제예 1. 정제의 제조Preparation Example 1. Preparation of tablets
실시예의 인삼 추출물........................100㎎Example Ginseng Extract 100 mg
전분.......................................................50mgStarch ................................................. ...... 50 mg
유당.......................................................50mgLactose ................................................. ...... 50 mg
스테아린산 마그네슘..............................5mgMagnesium stearate ............................. 5mg
상기의 성분들을 혼합하고 통상적인 정제의 제조방법에 따라 습식과립법 또는 건식과립법을 이용하여 정제를 제조하였다.The above components were mixed and tablets were prepared by a wet granulation method or a dry granulation method according to a conventional method for producing tablets.
제제예 2. 캡슐제의 제조Formulation Example 2. Preparation of capsules
실시예의 인삼 추출물........................100㎎Example Ginseng Extract 100 mg
전분.......................................................50mgStarch ................................................. ...... 50 mg
유당.......................................................50mgLactose ................................................. ...... 50 mg
탈크.......................................................1mgTalc ................................................. ...... 1 mg
스테아린산마그네슘............................. 5mgMagnesium stearate ............................. 5mg
상기의 성분들을 혼합하고 통상적인 캡슐제의 제조방법에 따라서 캡슐제를 제조하였다.The above components were mixed and capsules were prepared according to the conventional preparation method of capsules.
제제예 3. 액제의 제조Formulation Example 3. Preparation of a liquid preparation
실시예의 인삼 추출물...........................500㎎Example Ginseng Extract 500 mg
이성화당..................................................10ｇIsolation Party ................................................ ..10g
자당.........................................................10ｇsaccharose................................................. ........ 10g
레몬향.....................................................적당량Lemon flavor ................................................ ..... Appropriate amount
정제수를 가하여 전체..............................50㎖Purified water was added to the whole .............................. 50 ml
상기의 성분들을 통상적인 액제의 제조방법에 따라서 혼합하여 갈색병에 충전하고 멸균시켜서 액제를 제조하였다.The above components were mixed according to a conventional method for preparing a liquid preparation, filled in a brown bottle, and sterilized to prepare a liquid preparation.
제제예 4. 주사제의 제조Formulation Example 4. Preparation of injection
실시예의 인삼 추출물.............................50㎎Example Ginseng Extract 50 mg
소디움 메타비설파이트...........................2.6㎎Sodium Metabisulfite ........................... 2.6mg
메틸파라벤...............................................0.5㎎Methylparaben ............................................... 0.5 Mg
프로필파라벤...........................................0.1㎎Propylparaben ........................................... 0.1㎎
주사용 멸균증류수................................적당량Sterilized distilled water for injection ................................ Appropriate amount
상기의 성분들을 혼합하고 통상적인 방법으로 2 ml로 조절한 후, 앰플에 충전하고 멸균하여 주사제를 제조하였다.The above ingredients were mixed and adjusted to 2 ml in a conventional manner, filled in an ampoule and sterilized to prepare an injection.
제제예 5. 음료의 제조Formulation Example 5. Preparation of beverage
실시예의 인삼 추출물..........................100㎎Example Ginseng Extract 100 mg
비타민B2....................................................5㎎Vitamin B2 ................................................ .... 5 mg
비타민B6....................................................5㎎Vitamin B6 ................................................ .... 5 mg
비타민C...................................................80㎎Vitamin C ................................................ ... 80 mg
식이섬유..............................................1,000㎎Dietary fiber .............................................. 1000㎎
액상과당............................................10,000㎎Liquid fructose ............................................ 10,000㎎
유화제....................................................100㎎Emulsifier ................................................. ... 100 mg
레몬향......................................................50㎎Lemon flavor ................................................ ...... 50 mg
정제수....................................................적당량Purified water................................................. ... Appropriate amount
상기의 성분들을 혼합하여 용량이 50 ml가 되도록 하고, 혼합액을 3 μm의 필터로 통과시켜 여과한 최종 혼합액을 90 내지 93 ℃에서 20 초간 1차 살균하여 50 ml 병에 충진하고 80 내지 85 ℃에서 20 분간 2차 살균하여 음료제품을 제조하였다.The above components were mixed to make a volume of 50 ml, the mixture was passed through a 3-μm filter, and the resulting filtered mixture was first sterilized at 90 to 93 ° C for 20 seconds, filled in a 50 ml bottle, Followed by secondary sterilization for 20 minutes to prepare a beverage product.
제형예 1. 유연 화장수(스킨 로션)Formulation Example 1. Flexible lotion (skin lotion)
실시예의 인삼 추출물.......2.0Example Ginseng Extract 2.0
글리세린....................3.5Glycerin .................... 3.5
올레일알코올................1.5Oleyl alcohol ............... 1.5
에탄올........................5.5Ethanol ....................... 5.5
폴리솔베이트 80..............3.2Polysorbate 80 .............. 3.2
카르복실비닐폴리머............1.0Carboxyl vinyl polymer ............ 1.0
부틸렌 글리콜...............2.0Butylene glycol ............... 2.0
프로필렌 글리콜.............2.0Propylene glycol 2.0
방부제, 향료................적량Preservative, fragrance ................
정제수......................잔량Purified water ...................... Remaining amount
계...........................100Total ........................... 100
상기의 조성에 따라 통상적인 방법으로 유연 화장수를 제조하였다(단위 중량%).Flexible lotion was prepared according to the above composition in a conventional manner (unit weight%).
제형예 2. 영양 화장수(밀크 로션)Formulation Example 2. Nourishing lotion (milk lotion)
실시예의 인삼 추출물................2.0Ginseng Extract of the Example ... 2.0
글리세린............................3.0Glycerin ............................ 3.0
부틸렌글리콜.........................3.0Butylene glycol .......................... 3.0
프로필렌글리콜........................3.0Propylene glycol ........................ 3.0
카르복시비닐폴리머....................0.1Carboxyvinyl polymer 0.1
밀납..................................4.0Wax ................................. 4.0
폴리솔베이트 60........................1.5Polysorbate 60 ....................... 1.5
카프릴릭/카프릭 리글리세라이드..........5.0Caprylic / Capriculiglyceride .......... 5.0
스쿠알란................................5.0Squalane ................................ 5.0
솔비타세스퀴올레이트.....................1.5Sorbitac esquioleate ....................... 1.5
세테아릴 알코올..........................1.0Cetearyl Alcohol .......................... 1.0
트리에탄올아민...........................0.2Triethanolamine ........................... 0.2
방부제, 향료...........................적량Preservative, fragrance ............................
정제수.................................잔량Purified water ................................. Remaining amount
계......................................100Total ...................................... 100
상기의 조성에 따라 통상적인 방법으로 영양 화장수를 제조하였다(단위 중량%).Nutrition lotion was prepared according to the above composition in a conventional manner (unit weight%).
제형예 3. 영양 크림Formulation Example 3. Nourishing cream
실시예의 인삼 추출물.......................2.0Example Ginseng Extract 2.0
글리세린....................................3.5Glycerin .................................... 3.5
부틸렌글리콜................................3.0Butylene glycol ................................ 3.0
유동파라핀...................................7.0Liquid paraffin .......................................
베타글루칸..................................7.0Beta Glucan .................................. 7.0
카보머......................................0.1Carbomer ...................................... 0.1
카프릴릭/카프릭 리글리세라이드...........3.0Caprylic / capric wriglyceride ...................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................
스쿠알란.....................................5.0Squalane ..................................... 5.0
세테아릴 글루코사이드........................1.5Cetearyl Glucoside ....................... 1.5
소르비탄 스테아레이트.........................0.4Sorbitan stearate ......................... 0.4
폴리솔베이트 60...............................1.2Polysorbate 60 ............................... 1.2
트리에탄올아민.................................0.1Triethanolamine ................................. 0.1
방부제, 향료..................................적량Preservative, fragrance ..................................
정제수.........................................잔량Purified water ......................................... Remaining amount
계..............................................100Total .............................................. 100
상기의 조성에 따라 통상적인 방법으로 영양 크림을 제조하였다(단위 중량%).Nutritive creams were prepared according to the above composition in a conventional manner (unit weight%).
제형예 4. 팩Formulation Example 4. Pack
실시예의 인삼 추출물.............2.0Example Ginseng Extract 2.0
글리세린.........................4.0Glycerin ......................... 4.0
폴리비닐알콜.....................15.0Polyvinyl alcohol 15.0
히알루론산 추출물................5.0Hyaluronic acid extract ............... 5.0
베타글루칸.......................7.0Beta Glucan ....................... 7.0
알란토인..........................0.1Allantoin .......................... 0.1
노닐 페닐에테르...................0.4Nonyl phenyl ether
폴리솔베이트 60...................1.2Polysorbate 60 ................... 1.2
방부제, 향료......................적량Preservative, fragrance ......................
정제수.............................잔량Purified water .............................
계..................................100Total .................................. 100
상기의 조성에 따라 통상적인 방법으로 팩을 제조하였다(단위 중량%).A pack was prepared according to the above composition in a conventional manner (unit weight%).
제형예 5. 국소 투여용 약제(패취제)Formulation Example 5. Pharmaceutical preparation for local administration (patch)
실시예의 인삼 추출물.......................2.0Example Ginseng Extract 2.0
베타-1,3-글루칸............................3.0Beta-1,3-glucan ............................ 3.0
디에틸아민.................................0.7Diethylamine ................................. 0.7
아황산나트륨................................0.1Sodium Sulfite ................................ 0.1
폴리옥시에틸렌라우릴에테르(E.O=9)...........1.0Polyoxyethylene lauryl ether (EO = 9) ............ 1.0
폴리히드록시에틸렌세틸스테아릴에테르(Cetomacrogol 1000)......1.0Polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000) ... 1.0
점성의 파라핀 오일.........................2.5Viscous paraffin oil ....................... 2.5
카프릴산에스테르/카프르산에스테르(Cetiol LC).................2.5Caprylic acid ester / capric acid ester (Cetiol LC) 2.5
폴리에틸렌글리콜 400.......................3.0
폴리아크릴산(Carbopol 934P)................1.0Polyacrylic acid (Carbopol 934P) 1.0
정제수.....................................잔량Purified water .....................................
계..........................................100Total .......................................... 100
상기의 조성에 따라 통상적인 방법으로 국소 투여용 약제를 제조하였다(단위 중량%).According to the above composition, a medicament for topical administration was prepared by a conventional method (unit% by weight).
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.
실시예.Examples.
본 발명의 실시예에서는 표 1에 나열된 상업적으로 이용 가능한 효소와 4년근, 6년근 금산 인삼 및 4년근 풍기 인삼을 이용하였다.In the examples of the present invention, commercially available enzymes listed in Table 1 and 4-year-old roots, 6-year roots ginseng roots and 4-year roots roasted ginseng were used.
200 g의 인삼 뿌리를 잘게 썰어 인삼 슬러리를 제조하고, 인삼 슬러리를 혼합 그라인더를 이용하여 1 L의 멸균 증류수와 균질화하여 현탁액을 제조한 후, 표 2에 나열된 바와 같이 효소를 조합하여 당 분해효소로 12 내지 24 시간 동안 처리하였다. 당 분해효소 처리 후 효소 불활성 온도를 80 ℃, 100 ℃에서 30 분간, 121 ℃에서 15 분간 처리하여 비교하고, 250 ㎛의 체(sieve)로 압착여과한 후 48 내지 72 시간 동안 동결건조하여 농축시켰다.200 g of ginseng roots were finely chopped to prepare a ginseng slurry, and the ginseng slurry was homogenized with 1 L of sterilized distilled water using a mixed grinder to prepare a suspension. The enzymes were combined as shown in Table 2, For 12 to 24 hours. After the enzymatic digestion, the enzymatic inactivation temperature was treated at 80 캜, 100 캜 for 30 minutes, and 121 캜 for 15 minutes, followed by compression filtration with a 250 탆 sieve, lyophilization for 48 to 72 hours and concentration .
상기 표 2의 효소 단위에서, 효소의 최적 조건 하에서 30 ℃, 1 분당 1 μmol의 기질을 생성물로 변환시키기 위해 필요한 효소의 양을 1 단위(unit)라 하며, 이는 1 ml 중의 효소를 단위수로 효소의 농도를 표시한 것이다.In the enzyme unit of Table 2, the amount of enzyme required to convert 1 μmol of substrate to product at 30 ° C. per minute under optimal conditions of the enzyme is referred to as 1 unit, The concentration of the enzyme is indicated.
그 결과, 도 1 및 도 2에서 보는 바와 같이 아밀라아제 및 펙티나아제 처리(2500U + 1250U / 100 g 인삼 슬러리)시 Rg1과 Rb1 함량이 증대되었고, 도 3 및 도 4에서 보는 바와 같이 셀룰라아제를 고농도(2500U / 100 g 인삼 슬러리)로 24 시간 동안 처리할 경우 Rd 함량이 증대되었다.As a result, as shown in FIGS. 1 and 2, Rg1 and Rb1 contents were increased in amylase and pectinase treatment (2500 U + 1250 U / 100 g ginseng slurry), and as shown in FIG. 3 and FIG. 4, 2500 U / 100 g ginseng slurry) for 24 hours, the Rd content was increased.
또한, 비스코자임, 셀룰라아제 및 셀로비아제를 18, 24 시간 동안 처리(3.5 ml + 1000U + 1000U / 100 g 인삼 슬러리)할 경우 Rh1 함량이 증대(도 5 및 도 6 참조)되었으며, 48 시간 등 처리 시간이 길어질 경우 더욱 높은 Rh1 함량을 나타낼 것으로 보여진다. 셀룰라아제, 아밀라아제 및 셀로비아제를 36 시간 동안 처리(1000U + 1000U + 1000U / 100 g 인삼 슬러리)할 경우 화합물 K 함량이 증대되었다.In addition, when the viscose, cellulase and cellobiase were treated for 18, 24 hours (3.5 ml + 1000 U + 1000 U / 100 g ginseng slurry), the Rh content was increased (see FIGS. 5 and 6) The longer the time, the higher the Rh1 content. Compound K content was increased when cellulase, amylase and cellobiase were treated for 36 hours (1000 U + 1000 U + 1000 U / 100 g ginseng slurry).
실험예 1. 전체 사포닌(진세노사이드) 함량 측정Experimental Example 1. Measurement of total saponin (ginsenoside) content
추출된 전체 진세노사이드의 함량은 Wei, S.E., 2011(Isolation and determination of anti-nutritional compounds from root and shells of Peanut (Arachis hypogaea), Department of Chemical Science, Faculty of Science. Universiti Tunku Abdul Rahman, Selangor, Malaysia, p. 104.)에 개시된 방법에 따라 측정하였다.The content of total ginsenosides extracted was determined by the method of Wei, SE, 2011 (Isolation and determination of anti-nutritional compounds from root and shells of Peanut ( Arachis hypogaea ), Department of Chemical Science, Universiti Tunku Abdul Rahman, Selangor, Malaysia, p. 104.).
각각의 시료 100 ㎕에 농도가 80 %인 메탄올 수용액을 첨가하여 0.25 ml가 되도록 조절한 후, 농도가 8 %인 바닐린 시약 0.25 ml와 농도가 72 %인 황산 2.5 ml를 첨가하여 혼합하였다. 이후, 60 ℃의 수욕조에서 10 분간 처리하고 상온에서 냉각시켜 544 nm에서 흡광도를 측정하였다. 표준 곡선은 표준 사포닌으로서 다이오스게닌(diosgenin)의 다양한 농도를 이용하여 얻었고, 표준 사포닌 용액은 80 %의 메탄올 수용액을, 시약바탕시료(reagent blank)는 추출물이 함유된 시료 대신 80 %의 메탄올 수용액 0.25 ml를 이용하였다.A methanol aqueous solution having a concentration of 80% was added to each sample (100 μl) to adjust the volume to 0.25 ml. Then, 0.25 ml of vanillin reagent having a concentration of 8% and 2.5 ml of sulfuric acid having a concentration of 72% were added and mixed. Thereafter, the sample was treated in a water bath at 60 ° C for 10 minutes, cooled at room temperature, and absorbance was measured at 544 nm. The standard curve was obtained using various concentrations of diosgenin as a standard saponin, 80% methanol aqueous solution for standard saponin solution, reagent blank for 80% aqueous methanol solution instead of sample containing extract 0.25 ml was used.
실험예 2. 인삼 사포닌(진세노사이드)의 HPLC 분석Experimental Example 2. HPLC analysis of ginseng saponin (ginsenoside)
당 분해효소로 추출된 인삼 추출물을 같은 부피를 갖는 n-부탄올과 상온에서 20 분간 흔들어 혼합한 다음, 상 분리를 위해 5 분간 13,000 rpm으로 원심분리하여 부탄올 층을 모아 진공 증발기에서 건조시킨 후, HPLC 분석을 위해 메탄올에 용해시켰다.The resulting ginseng extract was mixed with n-butanol having the same volume for 20 minutes at room temperature, and the mixture was centrifuged at 13,000 rpm for 5 minutes for phase separation. The butanol layer was collected and dried in a vacuum evaporator, And dissolved in methanol for analysis.
HPLC 분석은 4.5 mm × 25 cm의 Sunfire C18 column을 이용하여 Waters HPLC system으로 수행되었고, HPLC용 아세토나이트릴과 물을 이동상으로 이용하고 1 ml/min의 이동상 흐름 속도로 수행되었다. 또한, 각각의 시료 20 μl를 C18 column에 주입하기 전에 0.25 μm PTFE 필터로 여과하였다. 다양한 진세노사이드의 용출이 203 nm에서 관찰되었고, 진세노사이드의 용출 순서와 양은 표준 진세노사이드 혼합물(Cerillant co., USA)의 크로마토그램과의 비교에 의해 산출되었다. 각각의 샘플에 대한 HPLC 분석은 정확성을 위해 세 번 반복하였다.HPLC analysis was performed on a 4.5 mm x 25 cm Sunfire C18 column using a Waters HPLC system and using acetonitrile for HPLC and water as mobile phase and at a flow rate of 1 ml / min. In addition, 20 μl of each sample was filtered through a 0.25 μm PTFE filter before being injected into the C18 column. The elution of the various ginsenosides was observed at 203 nm and the order and amount of elution of ginsenosides was calculated by comparison with the chromatogram of standard ginsenoside mixture (Cerillant co., USA). HPLC analysis for each sample was repeated three times for accuracy.
이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시태양일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다.
Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
Claims (14)
(2) 상기 현탁액에 아밀라아제(amylase) 및 펙티나아제(pectinase)를 첨가하여 20 내지 60 ℃에서 5 내지 80시간 동안 반응시키는 단계; 및
(3) 상기 반응 종료 후 70 내지 100℃에서 10 내지 50분 동안 효소를 불활성화시키는 단계;를 포함하여, 진세노사이드 Rg1 및 Rb1이 다량 함유된 인삼 추출물을 제조하는 것을 특징으로 하는 인삼 추출물의 제조방법.
(1) preparing a suspension by mixing at least one raw material selected from the group consisting of roots, stems, leaves and fruits of a plant of Panax species with a solvent;
(2) adding amylase and pectinase to the suspension and reacting at 20 to 60 ° C for 5 to 80 hours; And
(3) deactivating the enzyme at 70 to 100 ° C for 10 to 50 minutes after completion of the reaction, thereby producing a ginseng extract containing a large amount of ginsenoside Rg1 and Rb1 Way.
상기 (1)단계에서 용매는 물 및 탄소수 1 내지 4의 저급 알코올을 포함하는 군에서 선택된 1 이상인 것을 특징으로 하는 인삼 추출물의 제조방법.
The method according to claim 1,
Wherein the solvent in step (1) is at least one selected from the group consisting of water and a lower alcohol having 1 to 4 carbon atoms.
상기 (1)단계에서 현탁액은 용매 100 중량부에 대하여 인삼속 식물의 뿌리, 줄기, 잎 및 열매를 포함하는 군에서 선택되는 1 이상의 원료를 10 내지 25 중량부로 혼합한 것을 특징으로 하는 인삼 추출물의 제조방법.
The method according to claim 1,
In the step (1), the suspension is prepared by mixing 10 to 25 parts by weight of at least one raw material selected from the group consisting of root, stem, leaf and fruit of a plant of the genus Ginseng with respect to 100 parts by weight of the solvent. Gt;
상기 제조된 진세노사이드 Rg1 및 Rb1이 다량 함유된 인삼 추출물은 기능성 음료 조성물로 이용하는 것을 특징으로 하는 인삼 추출물의 제조방법.
The method according to claim 1,
Wherein the ginseng extract containing a large amount of the ginsenosides Rg1 and Rb1 is used as a functional beverage composition.
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