KR101056352B1 - Extraction method of donkey using high voltage pulse electric field - Google Patents
Extraction method of donkey using high voltage pulse electric field Download PDFInfo
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- ORHBXUUXSCNDEV-UHFFFAOYSA-N umbelliferone Chemical compound C1=CC(=O)OC2=CC(O)=CC=C21 ORHBXUUXSCNDEV-UHFFFAOYSA-N 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/232—Angelica
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
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Abstract
본 발명은 고전압 펄스 전기장 기술을 이용한 당귀의 효율적인 추출방법으로, 기존 일반 추출 방법으로는 유효성분을 쉽게 얻지 못한다는 점과, 고온 추출 시 활성 물질을 파괴할 수 있다는 점을 고려하여 선택한 당귀의 효율적인 추출방법에 관한 것이다. The present invention is an efficient extraction method of donkey using high voltage pulsed electric field technology, the effective extraction of the donkey using conventional extraction method, and considering the fact that the active material can be destroyed during high temperature extraction, It relates to an extraction method.
본 발명의 고전압 펄스 전기장 추출방법은 65~80℃에서 25㎸/㎝의 전기장을 5~20분간 2~3회 정도 주어 천연물인 당귀로부터 유효 성분을 뽑아내는 기술이다. 전기 에너지의 단기간 투여 시 활성 물질의 파괴 없이 세포벽만 선택적인 파괴가 가능해 목적하는 활성 물질의 생산성 증대 및 기타 유용 활성의 증진이 가능하다. The high voltage pulse electric field extraction method of the present invention is a technique of extracting an active ingredient from a natural donkey, giving an electric field of 25 mW / cm for 2 to 3 times at 5 to 20 minutes at 65 to 80 ° C. In the short-term administration of electrical energy, only the cell wall can be selectively destroyed without destroying the active substance, thereby increasing productivity of the active substance of interest and enhancing other useful activities.
본 발명은 종래 초고압 추출에 비하여 고가의 장비를 사용하지 않아도 된다는 점과, 시간을 절약할 수 있다는 점에서 특징을 가진다. 또한 수율과 생리활성 물질을 많이 내는 것으로 알려진 40~60kHz의 저에너지 초음파 추출과 비교했을 때 역시 비용과 시간 면에서 경제적이다. 본 발명의 고전압 펄스 전기장 추출방법을 통하여 얻어진 당귀 등의 천연물은 종래 통상적인 추출 기술에 비해 천연물의 생리활성 물질에 대한 높은 수율을 가진다.The present invention is characterized in that it does not need to use expensive equipment compared to the conventional ultra-high pressure extraction, and can save time. It is also economical in terms of cost and time when compared to 40-60 kHz low energy ultrasonic extraction, which is known for producing high yields and bioactive substances. Natural products such as Angelica, obtained through the high voltage pulsed electric field extraction method of the present invention have a higher yield for the bioactive materials of natural products than conventional extraction techniques.
당귀, 고전압 펄스 전기장 추출, 열수추출, 초음파추출, 초고압추출 Angelica, high voltage pulse electric field extraction, hot water extraction, ultrasonic extraction, ultra high pressure extraction
Description
본 발명은 고전압 펄스 전기장 기술을 이용한 천연물 당귀의 효율적인 추출방법에 관한 것이다. 더 상세하게 본 발명은 기존의 추출수율 향상을 위해 500~1000psi 이상에서 이루어지는 초고압 추출방법 및 40~60kHz에서 이루어지는 초음파 추출방법에서 발생했던 고비용과 많은 에너지 소비에 따른 단점을 개선하고자 고안한 것으로, 당귀의 열수추출물에 25-50㎸/㎝의 전기장을 가하여 수율 증진 효과와 면역생리활성 증진 효과를 갖는 추출물을 얻는 추출방법에 관한 것이다. 본 발명은 당귀의 면역 기능 증진과 생육 촉진 인자의 생산을 위해 추출물의 전처리 단계에서 고전압 펄스 전기장을 도입하여, 천연물 자체의 온도 상승을 수반하지 않고 신속히 유용물질의 생성을 유도하고, 전압의 연속처리가 가능하므로 물리적, 화학적 및 영양학적 특성을 펄스 처리 후에도 유지할 수 있는 장점을 가진다.The present invention relates to an efficient extraction method of natural product Angelica using high voltage pulse electric field technology. In more detail, the present invention is designed to improve the disadvantages of the high cost and high energy consumption that occurred in the ultra-high pressure extraction method made in 500 ~ 1000psi or more and the ultrasonic extraction method made in 40 ~ 60kHz to improve the conventional extraction yield, It relates to an extraction method of obtaining an extract having a yield enhancing effect and an immunophysiological activity enhancing effect by adding an electric field of 25-50㎸ / ㎝ to the hot water extract of. The present invention introduces a high-voltage pulse electric field in the pretreatment stage of the extract to enhance the immune function and production of growth factors of the Angelica gigas, induce the generation of useful substances quickly without involving the temperature rise of the natural product itself, and the continuous treatment of voltage It is possible to maintain physical, chemical and nutritional properties even after pulse treatment.
참당귀(Angelica gigas)는 미나리과에 속하는 다년생 초본으로 예부터 진정, 진통, 빈혈 및 월경통, 냉대하증 등의 부인과 질환에 보혈, 청혈약으로 널리 임상에 사용되어 왔다. 대한약전에 나오는 당귀의 약성은 '특이한 냄새가 나고 맛은 약간 쓰면서 달다'고 되어 있으나, 이는 중국당귀와 왜당귀에서만 느낄 수 있다. 참당귀는 단맛은 나지 않고 약간 쓴맛만 난다.Angelica gigas (Angelica gigas) is a perennial herb belonging to the Apiaceae family, and has been widely used as a blood donor and a blood medicinal agent for gynecological diseases such as sedation, analgesic, anemia and dysmenorrhea and hypothermia. The pharmacology of the tangs in the Korean Pharmacopoeia is said to have a unique smell and taste slightly sweet, but this can only be felt in Chinese donkeys and Koreans. Champagne don't taste sweet, only bitter.
성분으로는 coumarin 유도체인 decusin, decursinol과 umbelliferon 등이 보고되어 있으며, 최근 항돌연변이 활성, 암세포 생육억제활성, 간해독 활성 등이 보고되어 있다. 이러한 당귀는 최근 기능성 식품 소재의 원료로서 타블렛, 파우치 등으로 개발되고 있으며, 당귀의 새로운 이용에 대하여 많은 연구가 이루어지고 있다.The coumarin derivatives decusin, decursinol and umbelliferon have been reported. Recently, antimutagenic activity, cancer cell growth inhibitory activity, and liver detoxification activity have been reported. These donkeys are recently developed as tablets, pouches, and the like as raw materials for functional food materials, and many studies have been made on the new use of donkeys.
하지만 기존의 추출방법인 합성, 기타 천연물의 추출방법으로 얻은 당귀의 면역조절물질에 대해서 추출물의 활성에 비해 생산의 효율성이 낮은 것이 문제가 되었다. However, the efficiency of production compared to the activity of the extract for the immunomodulators of the Angelica Angelica obtained by the conventional extraction method synthesis, extraction of other natural products became a problem.
둘째, 기존에 추출 수율 향상을 위해 탄성체를 이루고 있는 질점들이 압축되었다가 팽창하면서 전파되는 파동의 주파수를 높여 극세 진동 작용을 통해 침투력 및 진동효과 증진을 이용하는 기술인 초음파를 이용한 추출 방법 역시 천연물의 높은 생리 활성, 면역 활성에 비해 시간과 비용이 커서 효율적이지 못했다. Second, the extraction method using ultrasonic wave, which is a technology that uses the enhancement of penetration force and vibration effect through the microscopic vibration, increases the frequency of waves propagated by compressing and expanding the existing points of elastic material to improve extraction yield. It was not efficient because of the time and cost compared to the activity and immune activity.
마지막으로 통상 100,000psi 이상의 압력을 가하여 세포벽 및 조직을 깨뜨려 성분의 용출을 가속화하고 단시간에 높은 용출량을 얻음으로써 효율을 높이는 방법인 초고압을 이용한 추출 방법은 높은 압력 공정으로 인하여 위험이 따르고 에너지를 발생시키기 위해 초고압을 도입하려다보니 그에 따른 장비의 고비용이 문제가 되었 다. Finally, the extraction method using ultra high pressure, which is a method of increasing the efficiency by accelerating the dissolution of the components by increasing the pressure of 100,000 psi or more and accelerating the dissolution of components and obtaining a high dissolution amount in a short time, is a risk and energy generation due to the high pressure process. In order to introduce ultra high pressure, the high cost of the equipment has become a problem.
따라서 본 발명은 천연 한약재 당귀의 효율적인 추출 및 저비용 장비로의 개선 효과 등을 고려해 고전압 펄스 전기장 추출을 고안했다. 짧은 시간에 65~80℃에서 25-50㎸/㎝의 전기장을 2회 정도 주어 천연물로부터 유효 성분을 뽑아내는 기술인 전기 펄스 법은 전기 에너지를 단기간 투여 시 활성 물질의 파괴 없이 세포벽만 선택적 파괴가 가능해져 목적하는 활성 물질의 생산성 증대 및 기타 유용 활성의 증진이 가능해 질 뿐 만 아니라 고온이 아니기 때문에, 당귀의 유효성분의 파괴 위험 또한 없으므로 안정적으로 추출물을 얻을 수 있다. 전압 펄스에 의한 당귀의 수율은 현탁액 속에 인가된 펄스의 수와 펄스 길이와 전계 강도에 의하여 결정된다. 또한 전기장의 세기와 처리 시간 이외에 펄스의 형태에 의해서도 영향을 받는다.Therefore, the present invention devised a high-voltage pulse electric field extraction in consideration of the effective extraction of natural herbal medicine Angelica Angelica and the improvement effect to low-cost equipment. The electric pulse method, which extracts active ingredient from natural products by giving 25-50㎸ / ㎝ electric field twice at 65 ~ 80 ℃ in a short time, can selectively destroy only the cell wall without destroying the active substance when short-term administration of electric energy. It is possible not only to increase the productivity of the desired active substance and to enhance other useful activities, but also because it is not high temperature, so there is no risk of destruction of the active ingredient of Angelica vulgaris, so that the extract can be stably obtained. The yield of the donkey by the voltage pulse is determined by the number of pulses applied in the suspension, the pulse length and the electric field intensity. It is also affected by the shape of the pulse in addition to the field strength and processing time.
본 발명의 고전압 펄스 전기장을 이용한 추출방법은 통상적으로 천연물의 전처리 단계에서 행해지고 있는 초고압 추출 공정 및 초음파 추출 공정에 비해 낮은 온도, 낮은 압력에서 추출함으로써 추출성분의 안정성을 높이고, 다양한 용매의 사용이 가능하면서도, 높은 수율을 가지는 것을 특징으로 한다. 따라서 많은 에너지가 소비되는 초고압 조건의 형성을 대신해 고전압 펄스를 줌으로써, 비용이 절감되는 효율적인 추출을 행할 수 있게 된다. 즉, 전기 펄스 추출법을 통해 초고압 기기의 경제적인 어려움을 극복할 수 있고, 지금까지의 단순한 추출방법으로 얻을 수 없는 새로운 생리활성 물질을 생산할 수 있게 한다.The extraction method using the high voltage pulse electric field of the present invention improves the stability of the extraction components by extracting at a low temperature and low pressure, compared to the ultra-high pressure extraction process and the ultrasonic extraction process that is usually performed in the pretreatment step of natural products, it is possible to use a variety of solvents Yet, it is characterized by having a high yield. Therefore, by giving a high voltage pulse in place of the formation of the ultra-high pressure condition that consumes a lot of energy, it is possible to perform an efficient extraction at which the cost is reduced. In other words, the electric pulse extraction method can overcome the economic difficulties of the ultra-high pressure device, it is possible to produce a new bioactive material that can not be obtained by the simple extraction method so far.
본 발명에 의한 고전압 펄스 전기장을 이용한 추출 방법은 종래 초고압 추출 조건에 비해 용이한 조건에서 고효율의 추출이 가능하게 함으로써, 종래 100,000psi 이상에서 행해지는 초고압 추출이 고압 및 고압에 의한 온도상승으로 추출성분이 불안정할 수 있으며 고압조건 형성을 위해 많은 에너지 및 고비용이 소요되는 단점을 가지고 있는 것을 개선한다. 또한 종래 초음파 추출이 장비의 고비용 및 추출 시간을 효율적으로 조절할 수 없는 단점을 가지고 있는 것을 개선한다. Extraction method using a high voltage pulse electric field according to the present invention enables the extraction of high efficiency under easy conditions compared to the conventional ultra-high pressure extraction conditions, the conventional ultra-high pressure extraction is carried out at a pressure rise of the high pressure and high pressure extracted at 100,000 psi or more This can be unstable and improves the disadvantage of having a lot of energy and cost to form high pressure conditions. It also improves that the conventional ultrasonic extraction has the disadvantage of not being able to efficiently control the high cost and extraction time of the equipment.
본 발명은 짧은 시간에 65~80℃에서 25-50㎸/㎝의 전기장을 2~3회 주어 천연물로부터 유효 성분을 뽑아내는 고전압 펄스 전기장 추출 방법을 도입함으로써 매우 짧은 시간에 추출을 완료할 수 있고, 전기 에너지를 단기간 투여 시 활성 물질의 파괴 없이 세포벽만 선택적 파괴가 가능해져 목적하는 활성 물질의 생산성 증대 및 기타 유용 활성의 증진이 가능해 질 뿐 만 아니라 저온에서 추출이 이루어지기 때문에, 뿌리 식물체인 당귀의 유효성분의 파괴 위험 또한 없으므로 안정적으로 추출물을 얻을 수 있다.The present invention is able to complete the extraction in a very short time by introducing a high voltage pulse electric field extraction method for extracting the active ingredient from natural products by giving an electric field of 25-50 ㎸ / ㎝ 2-3 times at 65 ~ 80 ℃ in a short time In addition, the short-term administration of electrical energy enables selective destruction of only the cell wall without destroying the active substance, thereby increasing the productivity of the desired active substance and enhancing other useful activities, as well as extracting at low temperatures. There is also no risk of destruction of the active ingredient, so the extract can be obtained stably.
따라서, 본 발명의 목적은 기존 일반추출법 및 이의 개선을 위해 도입된 초고압 추출방법 및 초음파추출방법의 조건형성이 용이하지 못하고 고비용의 설비 및 고에너지의 소비가 수반되는 것을 고려해서 보다 용이한 방법으로 높은 수율을 가지며 경제적으로 효율적인 추출밥법을 제공하는데 있다.Accordingly, an object of the present invention is to facilitate the formation of conditions of the ultra-high pressure extraction method and the ultra-high pressure extraction method and the ultrasonic extraction method introduced for the improvement of the existing general extraction method and easy to take into account the high cost of equipment and high energy consumption. To provide a high yield and economically efficient extraction rice method.
본 발명은 고전압 펄스 전기장 추출을 통한 당귀의 수율 및 생리활성을 측정하고, 종래의 일반 추출, 초음파 추출 및 초고압 추출 공정과 비교하여 그 차이를 알아보았다.The present invention measured the yield and physiological activity of the donkey through the extraction of high-voltage pulse electric field, and compared the conventional extraction, ultrasonic extraction and ultra-high pressure extraction process to find the difference.
상기 목적을 달성하기 위하여, 본 발명의 고전압 펄스 전기장을 이용한 당귀 추출방법은 당귀의 열수추출물에 25-50㎸/㎝의 전기장을 발생시키는 것을 특징으로 한다. In order to achieve the above object, the method for extracting donkey using the high voltage pulse electric field of the present invention is characterized in that it generates an electric field of 25-50 ㎸ / ㎝ to the hot water extract of the donkey.
또한, 상기 전기장 발생은 65-80℃에서 5-20분간 2-3회 발생시키는 것을 특징으로 한다.In addition, the electric field generation is characterized in that for 2-3 times 5-20 minutes at 65-80 ℃.
본 발명은 열수 추출된 뿌리 식물체 당귀 시료를 고전압 펄스 전기장 발생장치에서 입력 전원 교류전압을 high voltage transformer를 사용하여 전압을 올린 후 정류하였으며 완충효과를 위해 oil 속에 장치하였다. In the present invention, a hydrothermally extracted root plant Angelica sample was rectified after raising the voltage of the input power AC voltage using a high voltage transformer in a high voltage pulse electric field generator and installed in oil for a buffering effect.
본 발명은 특히, 처리 과정에서 열이 많이 발생하지 않기 때문에 천연물의 처리 과정에서 온도 상승이 없으므로 열에 의하여 야기될 수 있는 물질의 특성 변화, 발생물의 추가 등에 대한 문제가 없다. 그리고 고전압 펄스 전기장 기술은 전기 세포 조작이라고 하여 이미 세포 내로의 물질 도입법으로 이용되고 있기 때문에 본 발명은 유용한 추출방법으로 사용될 수 있으며 높은 생리활성 물질을 얻을 수 있다. In the present invention, since there is no heat generation during the treatment, there is no temperature rise during the treatment of the natural product, so there is no problem about the change in the properties of the material that can be caused by the heat and the addition of the generated product. In addition, since the high voltage pulse electric field technology is already used as a substance introduction method into a cell called electric cell manipulation, the present invention can be used as a useful extraction method and a high bioactive substance can be obtained.
본 발명의 고전압 펄스 전기장을 이용한 당귀 추출방법에서는 전기를 사용한 고전압을 이용하므로 종래 일반 추출, 초고압 추출, 초음파 추출법에 비하여 낮은 압력 및 온도에서 행해짐에 따라, 저에너지를 소비함으로써 공정비용이 적게 든다는 이점이 있다. 본 발명에서는 뿌리를 이용하는 천연물인 국내산 참당귀에 고전압펄스전기장을 가함으로써 수율을 증진시킬 수 있고, 면역 생리활성의 증진효과를 가진다.In the method of extracting a donkey using the high voltage pulse electric field of the present invention, since it uses a high voltage using electricity, it is performed at low pressure and temperature as compared to the conventional general extraction, ultra high pressure extraction, and ultrasonic extraction, and thus has the advantage of low process cost by consuming low energy. have. In the present invention, by adding a high-voltage pulse electric field to the domestic Angelica Angelica, a natural product using the root, it is possible to improve the yield, and has an effect of enhancing the immune physiological activity.
이하, 본 발명의 고전압 펄스 전기장을 이용한 뿌리 식물 당귀의 추출 방법에 대해 상세히 설명한다.Hereinafter, the extraction method of the root plant donkey using the high voltage pulse electric field of the present invention will be described in detail.
[제 1 공정 : 열수 추출 공정][1st process: hot water extraction process]
추출하고자 하는 생 또는 건조 국내산 참당귀(Angelica gigas)를 적당한 크기로 분쇄하고, 일정비(통상 10배)의 용매에서 60-85℃로 10-12시간 2회 연속 열수 추출한다. The raw or dried domestic Angelica gigas to be extracted is ground to an appropriate size, and extracted with hot water twice at a constant ratio (usually 10 times) for 10-12 hours at 60-85 ° C.
[제 2 공정 : 전기 펄스 공정]Second Process: Electric Pulse Process
제1공정에서 열수 추출된 시료를 고전압 펄스 전기장 발생기의 회분식으로 제작된 처리용기에 담는다. 본 발명에서는 고전압 펄스 전기장이 걸리는 부위의 전극 간격, 처리 시료의 부피, 펄스의 넓이, 펄스수(n)을 달리하여 뿌리 식물 당귀의 생리활성 물질 증가도를 향상시켰다. 고전압 펄스 전기장 발생기의 온도는 65~80℃로 25-50㎸/㎝의 전기장을 5~20분간 2~3회 주어 당귀의 세포벽이 깨어져 생리활성 물질이 나오도록 유도했다. The hydrothermally extracted sample in the first step is placed in a processing vessel manufactured batchwise of a high voltage pulsed electric field generator. In the present invention, the increase in the amount of physiologically active substances of the root plant Angelica was improved by varying the electrode spacing, the volume of the treated sample, the width of the pulse, and the number of pulses (n) of the site where the high voltage pulse electric field was applied. The temperature of the high-voltage pulse electric field generator was 65-80 ° C., and an electric field of 25-50 mA / cm was given two or three times for 5-20 minutes to induce the bio-active substance to break out of the cell wall of donkey.
[제 3 공정 : 추출물 수득 공정][Third step: extract obtaining step]
추출이 완료되면 고전압 펄스 전기장 추출물을 분리하고, 본 발명의 생리활성 실험을 위해 여과 및 감압농축 등으로 추출물을 분말 상태로 수득한다. 밸런스를 사용하여 분말상의 전기펄스 추출물의 수율을 얻는다. When the extraction is completed, the high voltage pulsed electric field extract is separated, and the extract is obtained in a powder state by filtration and concentration under reduced pressure for the physiological activity experiment of the present invention. The balance is used to obtain the yield of powdered electric pulse extract.
이하, 실시 예 및 실험 예를 통하여 본 발명을 더욱 상세히 설명한다.Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples.
실시 예 1. Example 1.
본 실험의 실시 예를 위하여 일반적으로 추출하여 식용 약용으로 널리 이용되는 국내산 참당귀를 가지고 시행하였다.For the example of this experiment was carried out with domestic Angelica gigas, which are generally extracted and used widely for edible medicinal purposes.
본 발명에서는 분쇄시켜 얻은 생 또는 건조 참당귀(건조중량 50g)에 10배(w/v)의 1차 증류수를 가하여 60℃에서 12시간 씩 2회 연속 열수 추출하였다. 열수 추출물을 고전압 펄스 전기장 발생기에서 회분식으로 제작된 처리용기에 담는다. 고전압 펄스 전기장 발생기에서 천연물의 생리활성 물질을 유도하기 위한 최적온도인 65℃의 온도가 유지되게 설정한다. 전기장의 설정은 25㎸/㎝, 전극간격은 4㎜, 저항은 400Ω, 펄스의 수는 30회, 펄스의 넓이는 100us, 펄스의 형태는 지수감쇄(expotential decay)파형으로 설정하여 5분동안 동일한 조건으로 2회 연속 반복한다. In the present invention, 10 times (w / v) of primary distilled water was added to the raw or dried Angelica gigas (dry weight 50g) obtained by pulverization, followed by continuous hot water extraction twice at 12O < 0 > C for 60 hours. The hydrothermal extract is placed in a batch prepared in a high voltage pulsed electric field generator. In the high voltage pulse electric field generator, the temperature of 65 ° C., which is an optimal temperature for inducing the bioactive substance of natural products, is maintained. The electric field is set to 25 mW / cm, electrode spacing is 4 mm, resistance is 400 ohms, the number of pulses is 30 times, the width of pulse is 100us, and the shape of pulse is exponential decay waveform. Repeat two times as a condition.
이후 전기 펄스 전처리를 마친 시료를 농축하였고, 동결 건조하여 분말화하였으며 수율을 확인하였다. After completion of the electric pulse pretreatment, the sample was concentrated, lyophilized and powdered, and the yield was confirmed.
실시 예 2. Example 2.
본 실험의 방법을 통한 추출 수율의 효율을 확인하기 위하여 국내산 참당귀를 대상으로 에탄올을 용매로 한 60℃ 열수추출, 100,000psi 초고압 공정 추출, 60kHz의 초음파 공정 추출의 세 가지 방법으로 추가 실험을 실행하였고 추출 효율을 비교하였다. 고전압 펄스 전기장 전처리를 실행한 추출물은 상기 실시 예1의 방법으로 수득한 추출물을 이용하였으며, 수율은 각 추출물을 감압 농축, 동결 건조하여 처음 시료에 대한 건조 중량(%)을 구하여 비교하였다.In order to verify the efficiency of extraction yield through the method of this experiment, three additional experiments were carried out for domestic Korean Angelica gigas: 60 ℃ hot water extraction using ethanol as a solvent, 100,000psi ultra-high pressure extraction, and 60kHz ultrasonic extraction. And extraction efficiency was compared. The extracts subjected to the high voltage pulse electric field pretreatment were used as extracts obtained by the method of Example 1, and the yields were compared by calculating the dry weight (%) for the first sample by concentrating the extract under reduced pressure and freeze drying.
표 1. 추출공정에 따른 추출수율 비교Table 1. Extraction yield comparison by extraction process
Ⅰ : 60℃ 열수추출, Ⅱ : 100,000psi 초고압추출, Ⅲ : 60kHz의 초음파추출, Ⅳ : 고전압펄스전기장추출Ⅰ: 60 ℃ hot water extraction Ⅱ: 100,000psi ultra high pressure extraction Ⅲ: 60kHz ultrasonic extraction Ⅳ: high voltage pulse electric field extraction
상기 표 1에서 보는 바와 같이 각 추출공정에 따른 수율은 참당귀가 60℃ 열수추출일 때 17.94%, 초고압추출일 때 24.26%, 초음파추출일 때 19.32%를 보이며, 본 발명에 의한 고전압펄스전기장 추출수율은 열수추출에 비해 2% 이상의 수율증진을 보였으며, 초고압 및 초음파 추출에 비해서는 수율은비슷하지만 수율대비 시 간과 비용 면에서는 주목할 수 있을 정도로 효율적이었다. As shown in Table 1, the yield according to each extraction process is 17.94% when the true donkey is 60 ℃ hot water extraction, 24.26% when ultra-high pressure extraction, 19.32% when ultrasonic extraction, high voltage pulse electric field extraction according to the present invention Yield was more than 2% higher than that of hot water extraction, and the yield was similar to ultra high pressure and ultrasonic extraction, but it was remarkably efficient in terms of time and cost.
실험 예 1.Experimental Example 1.
본 발명자는 본 발명으로 인한 추출물의 수율 증진을 통한 생리활성 증진 효과를 알아보기 위하여 실시 예 1 및 실시 예 2를 통해 얻어진 참당귀 추출물(증류수를 용매로 한 60℃ 열수추출물, 100,000psi 초고압추출물, 60kHz 초음파 추출물, 25㎸/㎝ 고전압 펄스 전기장 처리하여 수득한 추출물)을 이용하여 참당귀의 생리활성으로 알려진 면역 활성 실험을 실시하였다.The inventors of the present invention in order to determine the effect of improving the physiological activity through the increase in the yield of the extract according to the present invention (60 ℃ hot water extract, distilled water solvent, 100,000psi ultra-high pressure extract, Immune activity experiment known as physiological activity of True Angelica was performed using 60 kHz ultrasonic extract, 25 kHz / cm high voltage pulse electric field treatment).
본 실험에서는 0.5㎎/㎖의 시료의 첨가를 통한 인간 면역세포인 B세포와 T세포의 생육도를 측정하고 무처리한 대조군과 비교함으로써 시료의 면역 활성을 측정하였다.In this experiment, the immune activity of the samples was measured by measuring the growth of B cells and T cells, which are human immune cells, by adding 0.5 mg / ml of the sample and comparing it with the untreated control group.
표 2. 시료 첨가(0.5㎎/㎖)를 통한 참당귀 추출물의 생육조건 활성 측정Table 2. Determination of Growth Conditions Activity of Extracts of Angelica gigas Extracts by Sample Addition (0.5mg / mL)
(참당귀 추출물)sample
(Angelica Angelica Extract)
면역세포인 B세포, T세포 생육도의 경우 시료 투여 후 최고 수치를 기록한 6일 째의 생육정도를 표2에 나타내었다. 그 중 6일째 가장 높은 활성을 나타낸 추출물인 고전압펄스전기장의 경우 B세포와 T세포가 각각 14.61×104 cells/㎖, 15.24× 104 cells/㎖로 가장 높은 생육활성도를 나타났다. 상기 표 2의 실험결과를 통하여 60℃ 열수 추출물과 60kHz의 초음파 추출물 및 100,000psi의 초고압 추출물의 면역 활성은 큰 차이를 보이지 않는데 반해, 60℃ 열수 추출물과 비교했을 때 고전압펄스전기장 추출공정을 도입해 얻은 당귀 추출물의 경우 면역 활성이 탁월한 수치를 보임에 따라 생리활성에 있어서 고전압펄스전기장 추출공정이 면역 활성 증진 효과가 있음을 확인할 수 있었다. In the case of immune cell B-cell and T-cell growth, the growth of the highest day after the sample administration was shown in Table 2. In the high-voltage pulse electric field, which showed the highest activity on day 6, B cells and T cells showed the highest growth activity of 14.61 × 10 4 cells / ml and 15.24 × 10 4 cells / ml, respectively. While the immune activity of the 60 ℃ hot water extract, 60 kHz ultrasonic extract and 100,000 psi ultra-high pressure extract did not show a big difference, the high voltage pulse electric field extraction process was introduced in comparison with the 60 ℃ hot water extract. In the case of the obtained Angelica extract showed an excellent immune activity, it was confirmed that the high voltage pulse electric field extraction process in the physiological activity has an effect of enhancing the immune activity.
본 발명을 위한 실시 예 및 실험 예를 통해 수율과 면역세포 생육도 사이에 추가 비교를 할 수 있다. 100,000psi 초고압 추출물과 고전압펄스전기장 추출물의 수율이 각각 24.26±0.34(중량%), 20.20±0.25(중량%)인데 6일째 면역세포의 생육도 측정에서 고전압펄스전기장 추출물이 B세포와 T세포 각각 14.61×104 cells/㎖, 15.24×104 cells/㎖로 초고압 추출물의 B세포와 T세포 각각 14.56×104 cells/㎖, 14.02×104 cells/㎖보다 높은 활성을 나타내기 때문에 면역세포에 효과적으로 작용하는 추출물이 나왔음을 확인할 수 있었다. Examples and experimental examples for the present invention allow further comparison between yield and immune cell growth. The yield of 100,000 psi ultra high voltage extract and high voltage pulse electric field extract was 24.26 ± 0.34 (wt%) and 20.20 ± 0.25 (wt%), respectively. × 10 4 cells / mL and 15.24 × 10 4 cells / mL, which are more effective than immune cells because they show higher activity than B cells and T cells of 14.56 × 10 4 cells / mL and 14.02 × 10 4 cells / mL, respectively. It was confirmed that the extract was working.
실험 예 2.Experimental Example 2.
실시 예2에서의 60℃ 열수추출법, 60kHz 초음파 추출법 및 실시예1에서의 25㎸/㎝ 고전압펄스전기장 추출법을 통해 추출한 당귀를 HPLC를 이용해, decursin 함량을 측정하였고, decursin 함량정도를 나타낸 피크를 도1,2,3에 나타내었다. Decursin은 체내에서 혈액순환과 대사를 활성화시켜 항산화작용을 하는 생리활성 물질이다. 동일한 8min 시간대에서 60℃ 열수 추출물이 0.003~0.0035AU의 decursin 함량을 보인데 반해, 60kHz 초음파 추출물은 약 0.004AU의 함량을, 400Ω 고전압펄스전기장 추출물은 30% 이상 향상된 약 0.007AU의 decursin 함량을 보였다. The decursin content was measured by HPLC using the 60 ° C. hot water extraction method in Example 2, 60 kHz ultrasonic extraction method, and 25 kHz / cm high voltage pulse electric field extraction method in Example 1, and the decursin content was measured. 1,2,3 is shown. Decursin is a bioactive substance that acts as an antioxidant by activating blood circulation and metabolism in the body. At the same 8min time, 60 ℃ hydrothermal extract showed decursin content of 0.003 ~ 0.0035AU, while 60kHz ultrasonic extract showed about 0.004AU content and 400Ω high voltage pulsed electric field extract showed decursin content of about 0.007AU improved by more than 30%. .
도1은 열수추출법에 따른 Decursin 함량을 나타낸 그래프이다.1 is a graph showing the Decursin content according to the hot water extraction method.
도2는 초음파추출법에 따른 Decursin 함량을 나타낸 그래프이다.Figure 2 is a graph showing the Decursin content according to the ultrasonic extraction method.
도3은 고전압 펄스 전기장 추출에 따른 Decursin 함량을 나타낸 그래프이다.Figure 3 is a graph showing the Decursin content according to the extraction of high voltage pulse electric field.
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