KR100758814B1 - A simplified purificating method of bee venom - Google Patents
A simplified purificating method of bee venom Download PDFInfo
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- KR100758814B1 KR100758814B1 KR1020060108852A KR20060108852A KR100758814B1 KR 100758814 B1 KR100758814 B1 KR 100758814B1 KR 1020060108852 A KR1020060108852 A KR 1020060108852A KR 20060108852 A KR20060108852 A KR 20060108852A KR 100758814 B1 KR100758814 B1 KR 100758814B1
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- A23L3/40—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by drying or kilning; Subsequent reconstitution
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Abstract
Description
도1은 본 발명의 실시예에 따른 정제 방법의 흐름도이다.1 is a flowchart of a purification method according to an embodiment of the present invention.
도2는 시판되는 봉독과 본 발명의 실시예에 따른 봉독의 액체크로마토그래피 그래프이다.2 is a liquid chromatography graph of bee venom and bee venom according to an embodiment of the present invention.
본 발명은 봉독의 간이 정제 방법에 관한 것으로, 보다 상세하게는 양봉장에서 봉독 채집 장치를 사용하여 채취한 봉독으로부터 순수한 봉독만을 성분의 변화 없이 간편하게 정제하기 위한 방법에 관한 것이다.The present invention relates to a simple purification method of bee venom, and more particularly to a method for easily purifying only pure bee venom from bee venom collected using a bee venom collection device in apiary farms without changing the ingredients.
봉독이란 벌이 가진 독을 말한다. 이와 같은 봉독은 치료의 목적을 위해 널리 사용되었는데, 기원전 4세기경 히포크라테스가 벌침을 사용했다는 기록이 있으며, 이슬람 경전인 코란에도 벌침을 치료 목적으로 사용한 기록이 있고, 대한민국에서도 예로부터 민간요법의 일종으로 벌침을 치료에 응용하였다.Bee venom is the poison that bees have. Such bee venom was widely used for therapeutic purposes, including the record of Hippocrates' use of acupuncture around the 4th century BC, and the Koran, an Islamic scripture, for the purpose of treating bee venom. Bee was applied to the treatment.
그러나 과거에 사용된 봉독의 활용은 벌을 잡아 벌침을 환부에 주입하거나, 벌침만을 뽑아 낸 뒤 이를 환부에 주입하였다. 그러나 이와 같은 방법은 봉독의 일정한 양이나 농도를 조절하기가 쉽지 않아 치료의 객관화 및 일반화가 어려워 쇼크 등의 부작용이 발생할 수 있다.However, the use of bee venom used in the past has caught a bee and injected the bee sting into the affected area, or extracted only the bee sting and injected it into the affected area. However, this method is difficult to control the constant amount or concentration of bee venom, so it is difficult to objectify and generalize the treatment may cause side effects such as shock.
한편, 봉독액을 이용한 치료는 환자의 상태와 체질에 따라 주입하는 약의 농도 및 용량이 객관적으로 조절 가능하여 거의 부작용이 없는 안전한 치료방법이 될 수 있다. 이는 미국이나 서구 유럽에서 이미 사용하고 있는 방법으로 대한민국은 봉독 채집이 가능하지 않아 90년대 초 미국 식품의약국의 승인을 얻어 미국으로부터 수입된 봉독을 사용하여 아피톡신이라는 제품이 생산되어 각종 염증 및 통증 치료에 이용하고 있었으나, 최근에는 살아있는 꿀벌로부터 다량의 봉독을 채취할 수 있는 봉독 채집 장치의 보급으로 양봉 농가에서도 자체적으로 봉독을 채취하는데 성공하였다. 그러나 봉독 채집 장치로부터 채취한 봉독은 야외 양봉장에서 채취하기 때문에 먼지나 흙과 같은 불순물이 다양 혼입되어 있으며, 꿀벌의 몸에 묻어 있던 벌꿀, 화분, 프로폴리스와 같은 이물질이 함께 채취된다. 이와 같은 이물질은 봉독의 성분 변화를 야기하여 봉독의 생리 활성능을 저하시킬 수 있다.On the other hand, treatment with bee venom solution can be objectively controlled by the concentration and dose of the drug to be injected according to the condition and constitution of the patient can be a safe treatment method with almost no side effects. This method is already used in the United States and Western Europe. Korea cannot collect bee venom, so it was produced by the US Food and Drug Administration in the early 90's, and a product called apitoxin was produced from bee venom. Although it has been used for treatment, recently, bee farms have successfully collected bee venom by supplying a bee venom collecting device that can collect a large amount of bee venom from live bees. However, the bee venom collected from the bee venom collection device is collected in an outdoor apiary and contains various impurities such as dust and soil, and foreign substances such as honey, pollen, and propolis that are on the body of the bee are collected together. Such foreign matter may cause a change in the components of bee venom, thereby lowering the physiological activity of bee venom.
본 발명은 전술된 종래 기술에 따른 문제점을 해결하기 위하여 도출된 것으로, 봉독 채집 장치를 사용하여 채취한 봉독을 손쉽게 정제할 수 있을 뿐만 아니라, 봉독의 성분 변화를 최대한 억제할 수 있는 정제 방법의 제공을 목적으로 한다.The present invention is derived to solve the problems according to the prior art described above, not only can be easily purified bee venom collected using a bee venom collecting device, but also provides a purification method that can suppress the change in the components of bee venom to the maximum For the purpose.
전술된 목적을 달성하기 위하여, 본 발명의 실시형태에 따른 봉독의 간이 정제 방법은, 봉독 채집 장치의 유리 채집판에서 수거한 봉독을 정제하는 방법에 있 어서, 수거된 봉독을 멸균 증류수에서 5 내지 15분간 용해시키는 제1단계; 상기 제1단계에 의한 용액을 5 내지 15분간 원심 분리하는 제2단계; 상기 제2단계에서 원심 분리된 용액 중 상청액을 여과지에서 여과하는 제3단계; 및 상기 제3단계에서 여과된 여과액을 동결 건조하는 제4단계;를 포함하는 것을 특징으로 한다.In order to achieve the above object, the simple purification method of bee venom according to the embodiment of the present invention, in the method of purifying the bee venom collected from the glass collecting plate of the bee venom collecting device, the collected bee venom in the sterile distilled water 5 to A first step of dissolving for 15 minutes; A second step of centrifuging the solution of the first step for 5 to 15 minutes; A third step of filtering the supernatant in the solution centrifuged in the second step on a filter paper; And a fourth step of freeze-drying the filtrate filtered in the third step.
본 발명에서 상기 제1단계는: 수거된 봉독 무게의 45 내지 55배의 3차 멸균 증류수가 사용될 수 있다.The first step in the present invention: 45 to 55 times the weight of the collected bee venom can be used as sterile distilled water.
본 발명에서 상기 제2단계는: 4℃, 3000g에서 원심 분리가 수행될 수 있다.The second step in the present invention: centrifugation may be performed at 4 ℃, 3000g.
본 발명에서 상기 제3단계는: 8㎛의 기공 크기를 갖는 여과지에서 여과될 수 있다.In the present invention, the third step may be filtered on a filter paper having a pore size of 8 μm.
본 발명에서 상기 제3단계는: 진공 펌프를 사용하여 강제 여과할 수 있다.In the present invention, the third step may be: forced filtration using a vacuum pump.
본 발명에서 상기 제4단계는: -65 내지 -75℃의 온도에서 급속 동결된 여과액을 건조시킬 수 있다.In the present invention, the fourth step may be: drying the rapidly frozen filtrate at a temperature of -65 to -75 ° C.
이하에서, 첨부된 도면들을 참조하여 본 발명의 실시예가 기술된다.In the following, embodiments of the present invention are described with reference to the accompanying drawings.
하기에서 본 발명을 설명함에 있어서, 관련된 공지 기능 또는 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우에는 그 상세한 설명은 생략될 것이다. 또한 후술되는 용어들은 본 발명에서의 기능을 고려하여 설정된 용어들로서 이 용어들은 제품을 생산하는 생산자의 의도 또는 관례에 따라 달라질 수 있으며, 용어들의 정의는 본 명세서 전반에 걸친 내용을 토대로 내려져야 할 것이다.In the following description of the present invention, if it is determined that detailed descriptions of related known functions or configurations may unnecessarily obscure the subject matter of the present invention, the detailed description will be omitted. In addition, the terms to be described later are terms set in consideration of functions in the present invention, and these terms may vary according to the intention or custom of the producer producing the product, and the definition of the terms should be made based on the contents throughout the present specification.
(실시예)(Example)
이하에서 첨부된 도면 도1과 도2를 참조로 본 발명의 실시예에 따른 봉독의 간이 정제 방법을 설명한다.Hereinafter, a simple purification method of bee venom according to an embodiment of the present invention will be described with reference to FIGS. 1 and 2.
도1은 본 발명의 실시예에 따른 정제 방법의 흐름도이다.1 is a flowchart of a purification method according to an embodiment of the present invention.
도2는 시판되는 봉독과 본 발명의 실시예에 따른 봉독의 액체크로마토그래피 그래프이다.2 is a liquid chromatography graph of bee venom and bee venom according to an embodiment of the present invention.
도1에 도시된 바와 같이, 본 발명의 실시예에 따른 봉독의 간이 정제 방법(이하에서 설명의 편의를 위하여, ‘봉독의 간이 정제 방법’은 간략히 ‘정제 방법’이라 한다)은 용해(S10), 원심 분리(S20), 여과(S30) 및 동결건조(S40)를 포함하여 이루어진다.As shown in Figure 1, a simple purification method of bee venom according to an embodiment of the present invention (for convenience of description below, 'simple purification method of bee venom' is briefly referred to as 'purification method') is dissolved (S10) , Centrifugation (S20), filtration (S30) and lyophilization (S40).
제1단계는 용해 단계로, 봉독 채집 장치의 유리 채집판에서 수거한 봉독을 멸균 증류수에서 5 내지 15분간, 바람직하게는 10분간 용해시킨다(S10). 이때에 사용되는 멸균 증류수는 3차 멸균 증류수로 수거된 봉독 무게의 45 내지 55배, 바람직하게는 50배의 양이 사용된다.The first step is a dissolution step, the bee venom collected from the glass collection plate of the bee venom collecting device is dissolved for 5 to 15 minutes, preferably 10 minutes in sterile distilled water (S10). The sterile distilled water used at this time is used in an amount of 45 to 55 times, preferably 50 times the weight of bee venom collected by the third sterile distilled water.
제2단계는 원심 분리 단계로, 제1단계에 의한 용액을 5 내지 15분간, 바람직하게는 10분간 원심 분리한다(S20). 이때의 원심 분리 조건은 4℃, 3000g에서 수행되는 것이 바람직하다. 이와 같은 원심 분리에 의해 가장 위층엔 왁스와 프로폴리 스 등의 지질층이 얇게 막으로 형성되고, 바닥에는 모래, 흙 및 먼지 등이 쌓이게 된다.The second step is a centrifugation step, the solution of the first step is centrifuged for 5 to 15 minutes, preferably 10 minutes (S20). The centrifugal conditions at this time is preferably carried out at 4 ℃, 3000g. By such centrifugation, a lipid layer such as wax and propolis is formed in a thin film on the top layer, and sand, soil and dust are accumulated on the bottom.
제3단계는 여과 단계로, 제2단계에서 원심 분리된 용액 중 상청액을 여과지에서 여과한다(S30). 이때의 여과 단계는, 먼저 일반 여과지로 펠릿(pellet)이 떨어지지 않도록 여과시킨 뒤, 미세 물질의 여과를 위해 8㎛의 기공 크기를 갖는 여과지에서 여과 하는 것이 바람직하다. 또한 봉독 양이 많을 경우에는 진공 펌프를 사용하여 강제 여과할 수도 있다. 이상의 여과 단계를 통해, 순수한 봉독 성분을 갖는 여과액을 얻을 수 있게 된다.The third step is a filtration step, and the supernatant in the solution centrifuged in the second step is filtered on a filter paper (S30). In this case, the filtration step, first filter the pellets (pellets) so as not to fall into the general filter paper, it is preferable to filter on a filter paper having a pore size of 8㎛ for filtration of the fine material. In addition, if the amount of bee venom is high, a vacuum pump may be used for forced filtration. Through the above filtration step, it is possible to obtain a filtrate having a pure bee venom component.
제4단계는 동결 건조 단계로, 제3단계에서 여과된 여과액을 동결 건조한다(S40). 이때의 동결 건조 조건은 -65 내지 -75℃의 온도에서 급속 동결된 여과액을 동결건조기를 사용하여 건조시키는 것이 바람직하다.The fourth step is a freeze drying step, the freeze-dried filtrate filtered in the third step (S40). The freeze-drying condition at this time is preferably dried by using a lyophilizer rapidly freezing the filtrate at a temperature of -65 to -75 ℃.
이상의 단계를 통해, 정제된 봉독이 준비된다. 이와 같은 단계를 통해 정제된 봉독을 액체크로마토그래피를 사용하여 분석하면, 도2에 도시된 바와 같은 그래프를 얻을 수 있다.Through the above steps, purified bee venom is prepared. If the purified bee venom is analyzed using liquid chromatography through this step, a graph as shown in FIG. 2 may be obtained.
도2에 도시된 바와 같이, 본 발명의 실시예에 따라 제조된 봉독(B)은 시판중인 봉독(A)과 비교할 때, 봉독의 주요 성분인 멜리틴(M: Melittin), 아파민(A: Apamin) 및 포스포리파아제 A2(PA2: Phospholipase A2)의 성분이 거의 동일함을 알 수 있다.As shown in Figure 2, the bee venom (B) prepared according to an embodiment of the present invention, when compared to commercial bee venom (A), the main components of bee venom (M: Melittin), apamin (A: It can be seen that the components of Apamin) and phospholipase A2 (PA2: Phospholipase A2) are almost identical.
이상으로 본 발명의 실시예를 첨부된 도면을 참조로 기술하였다. 그러나 본 발명은 전술된 실시예에만 특별히 한정되는 것은 아니며, 필요에 따라, 당업자에 의해, 첨부된 청구범위의 정신과 사상 내에서 다양한 수정 및 변경이 가능함에 유의해야 한다.The embodiments of the present invention have been described above with reference to the accompanying drawings. However, it is to be noted that the present invention is not particularly limited only to the above-described embodiments, and that various modifications and changes can be made by those skilled in the art within the spirit and spirit of the appended claims as necessary.
전술된 바와 같이, 본 발명에 따르면, 양봉농가에서 봉독을 채취한 뒤 봉독의 성분 변화 없이 봉독 이외의 불순물 등을 제거할 수 있다.As described above, according to the present invention, after collecting bee venom in a bee farm, impurities other than bee venom may be removed without changing the bee venom component.
이에 따라, 봉독을 장기간 보관하더라도 성분 변화를 최소화할 수 있어, 봉독의 지속적인 공급이 가능하고, 봉독을 원료로 하는 식의약품 개발에 유용하게 사용될 수 있다.Accordingly, even if the bee venom is stored for a long time, it is possible to minimize the change in ingredients, it is possible to continuously supply bee venom, it can be usefully used in the development of food and pharmaceuticals using bee venom as a raw material.
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Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
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KR101150293B1 (en) | 2008-09-03 | 2012-06-08 | 서영철 | Manufacturing process of Extract of Bee Venom for food additive |
CN102526116A (en) * | 2011-11-23 | 2012-07-04 | 程文显 | Method for refining bee venom |
WO2013103178A1 (en) * | 2012-01-04 | 2013-07-11 | Republic Of Korea(Management : Rural Development Administration) | Method for purifying bee venom on mass scale |
WO2014003222A1 (en) * | 2012-06-29 | 2014-01-03 | 주식회사 청진바이오텍 | Method for isolating the active component of bee venom |
KR101359528B1 (en) * | 2011-09-22 | 2014-02-27 | 대한민국 | Radiation protective composition comprising effective bee-venom and royal jelly |
KR101368341B1 (en) | 2012-12-28 | 2014-03-03 | 전지헌 | Method for collecting bee venom |
KR20160043167A (en) | 2014-10-10 | 2016-04-21 | 대한민국(농촌진흥청장) | Method for seperating bee venom containing active amines and food composition thereof |
KR102028960B1 (en) * | 2019-06-28 | 2019-11-08 | 유바이오주식회사 | Method for preparing purified bee venom |
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Cited By (15)
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KR101150293B1 (en) | 2008-09-03 | 2012-06-08 | 서영철 | Manufacturing process of Extract of Bee Venom for food additive |
KR101359528B1 (en) * | 2011-09-22 | 2014-02-27 | 대한민국 | Radiation protective composition comprising effective bee-venom and royal jelly |
CN102526116A (en) * | 2011-11-23 | 2012-07-04 | 程文显 | Method for refining bee venom |
CN102526116B (en) * | 2011-11-23 | 2013-08-07 | 程文显 | Method for refining bee venom |
US20140314871A1 (en) * | 2012-01-04 | 2014-10-23 | Republic Of Korea (Management:Rural Development Administration) | Method for purifying bee venom on mass scale |
KR101382404B1 (en) * | 2012-01-04 | 2014-04-14 | 대한민국 | Massive purification method of bee venom |
CN104023556A (en) * | 2012-01-04 | 2014-09-03 | 韩国农村振兴厅 | Method For Purifying Bee Venom On Mass Scale |
WO2013103178A1 (en) * | 2012-01-04 | 2013-07-11 | Republic Of Korea(Management : Rural Development Administration) | Method for purifying bee venom on mass scale |
EP2800477A4 (en) * | 2012-01-04 | 2015-07-01 | Republic Korea Man Rural Dev | Method for purifying bee venom on mass scale |
US9233129B2 (en) * | 2012-01-04 | 2016-01-12 | Republic Of Korea (Management: Rural Development Administration) | Method for purifying bee venom on mass scale |
WO2014003222A1 (en) * | 2012-06-29 | 2014-01-03 | 주식회사 청진바이오텍 | Method for isolating the active component of bee venom |
KR101368341B1 (en) | 2012-12-28 | 2014-03-03 | 전지헌 | Method for collecting bee venom |
KR20160043167A (en) | 2014-10-10 | 2016-04-21 | 대한민국(농촌진흥청장) | Method for seperating bee venom containing active amines and food composition thereof |
KR101704123B1 (en) | 2014-10-10 | 2017-02-20 | 대한민국(관리청: 특허청장, 승계청: 농촌진흥청장) | Method for seperating bee venom containing active amines and food composition thereof |
KR102028960B1 (en) * | 2019-06-28 | 2019-11-08 | 유바이오주식회사 | Method for preparing purified bee venom |
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