KR20120036565A - K w-100 and stevia extract containing disinfectant manufacturing method - Google Patents
K w-100 and stevia extract containing disinfectant manufacturing method Download PDFInfo
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Abstract
Description
본 발명은 K W-100과 스테비아 추출물이 함유된 소독제의 제조 방법에 관한 것이다.The present invention relates to a method for preparing a disinfectant containing K W-100 and stevia extract.
스테비아 추출물과 K W-100의 각각의 적정 농도를 조합하여 예방용 항바이러스 소독제 제품 개발하여 계사?돈사?우사?어류양식장에서의 예방효과가 있는 농도 와 실제 소독제의 효능이 나타날 수 있는 적정 농도를 조사하여 최적 후보농도 2가지를 이용하여 효능을 만족하는 최적농도를 조합한 후 제품을 생산한다.Developing anti-viral disinfectant products by combining the appropriate concentrations of stevia extract and K W-100, it is possible to determine the concentrations that are effective in the housing, pigs, barns and fish farms and that the effective disinfectant can be shown. The product is produced after combining the optimal concentrations that satisfy the efficacy by using two optimal candidate concentrations.
본 발명은 K W-100과 스테비아 추출물을 활용하여 만든 소독제가 물리화학적인 특성에 따라 각종 가용화 기술을 이용하여 용해도를 증가시키며 빠른 흡수율을 달성하기 위한 방법으로 액상의 소독제를 제조하여 면역력을 향상시킴을 목적으로 한다.The present invention is a disinfectant made by using K W-100 and stevia extract to increase the solubility using various solubilization technology according to the physicochemical properties and to improve the immunity by preparing a liquid disinfectant in a way to achieve a fast absorption rate For the purpose.
본 발명은 스테비아의 추출 정제기술을 활용하여 K W-100 및 후보물질을 이용한 가금류 항 바이러스 소독제 제품을 개발하고 소독제의 제형은 가금 및 양돈에 가장 쉽게 예방 및 치료 효과를 극대화 할 수 있는 제형으로 우선 선정함에 있다.The present invention develops a poultry antiviral disinfectant product using K W-100 and a candidate substance using the extraction and purification technology of stevia, and the formulation of the disinfectant is the first to maximize the prevention and treatment effect on poultry and pigs. In the selection.
본 발명은 스테비아의 추출 정제기술을 활용하여 K W-100 및 후보물질을 이용한 가금류 항바이러스 소독제 제품을 개발함에 있다. 가금 인플루엔자 예방용 소독제 제형은 가금 및 양돈에 가장 쉽게 예방 및 치료 효과를 극대화 할 수 있는 제형으로 우선 선정한다. The present invention is to develop a poultry antiviral disinfectant product using K W-100 and candidate substances using the extraction and purification technology of stevia. Poultry influenza preventive disinfectant formulations are first selected as the formulations that will maximize the most prophylactic and therapeutic effects on poultry and swine.
도 1은 소독제의 제조 공정도.1 is a manufacturing process diagram of the disinfectant.
본 발명은 바이러스를 효과적으로 예방 및 치료할 수 있는 제품의 개발에 관한 것으로 개발의 범주는 동물사료용 첨가제 및 소독 제뿐만 아니라 식품, 의약품, 의약 외 품, 동물용 의약품 등이 해당된다.양계 사, 돈사 및 축사, 어류양식장에 가축질병의 예방을 위한 제품으로는 생균제 및 소독제 등이 있으며 이들 제품은 동물용으로 사용될 수 있다.The present invention relates to the development of a product that can effectively prevent and treat viruses, the scope of development is not only additives and disinfectants for animal feed, but also food, pharmaceuticals, quasi-drugs, animal medicines and the like. Products for the prevention of livestock diseases in barns and fish farms include probiotics and disinfectants. These products can be used for animals.
바이러스의 소독제 효력시험Disinfectant Effect Test of Virus
1.공시 바이러스1.Disclosure Virus
효력시험에 사용하는 공시 바이러스주는 성상 및 출처를 입증할 수 있어야 한다 Disclosure virus lines used for efficacy testing should be able to verify their identity and source.
2.세포주 등2. Cell line, etc.
바이러스 종류의 생물학적 특성에 따라 적합한 세포, 발육란 또는 실험동물 등을 시험에 사용한다. Appropriate cells, embryonated eggs or laboratory animals should be used for testing depending on the biological characteristics of the virus type.
3.배지시약 등3.Medium reagent
선정된 세포증식에 적합한 배지와 관련시약을 사용하며, 증류수희석액, 경수희석액 및 유기물 희석액은 세균의 시험법에 준한다. 단, 유기물 희석액에는 효모추출액 대신에 5% 소태아혈청을 사용한다. Medium and related reagents suitable for the selected cell proliferation shall be used. Distilled water diluent, hard water diluent and organic diluent shall follow the bacterial test method. However, 5% fetal bovine serum is used instead of yeast extract in organic diluent.
소독제의 약품성분 중화배지는 사용세포에 적합한 배지에 불활화된 10% 소태아혈청을 함유한 배지를 사용한다. Drug neutralization medium of the disinfectant is a medium containing 10% fetal bovine serum inactivated in a medium suitable for the cells used.
4.바이러스 소독제 효력시험 4. Virus disinfectant efficacy test
(바이러스 배양) 계대배양 중의 활력있는 바이러스를 사용하되 바이러스의 증식이 최대인 싯점에서 채독하여 사용직전까지 단시간 동안 얼음물에서 보관한다. (Virus cultivation) Use a viable virus in the subculture, but read it at the point of maximum virus growth and store it in ice water for a short time until just before use.
(소독제의 희석) 소독제는 예상 희석배수가 500배 이하일 경우는 50, 100, 150, 200, 250, 300, 400, 500 등으로 희석하되, 그 이상의 예상 희석배수 농도에서는 달리할 수 있다. 단, 정밀한 희석배수가 요구되는 경우는 세균 등의 소독제 효력시험3의항의 방법에 따른다. (Dilution of disinfectant) Disinfectants can be diluted to 50, 100, 150, 200, 250, 300, 400, 500, etc., if the expected dilution is 500 times or less, but may be different at higher expected dilution concentrations. However, if precise dilution drainage is required, follow the method in paragraph 3 of disinfectant efficacy test for bacteria.
(소독제 반응) 위의 항에서 증식된 4의 바이러스액 1.0 ml를 4의 5%의 유기물희석액 19.0 ml에 섞은 후에, 1분 간격으로 혼합액 2.5ml를 꺼내어 4에 있는 동량의 소독제 희석액이 들어 있는 시험관에 넣고 혼합한 다음, 4에서 정확히 30분간 반응을 시키며 도중에 10분마다 혼합하여 준다. (Disinfectant Reaction) After mixing 1.0 ml of the virus solution 4 grown in the above section into 19.0 ml of 4% 5% organic diluent solution, 2.5 ml of the mixture solution was removed at 1 minute intervals, and the test tube containing the same amount of disinfectant diluent in 4 was added. After mixing, react at 4 minutes for exactly 30 minutes and mix every 10 minutes along the way.
단, 발육란을 이용한 시험에서는 요막강액(allantoic fluid) 1.0 ml를 24.0ml의 유기물희석액에 넣은 혼합물과 소독액을 동량 혼합(1:1)한다.However, in the test using embryonated eggs, 1.0 ml of allantoic fluid was added to 24.0 ml of organic diluent and the same amount of disinfectant was mixed (1: 1).
(중화반응) 소독제의 반응이 끝나면 소독제의 효능을 중화하기 위하여 즉시 1.0ml를 꺼내어 37의 동량의 중화배지에 넣고 혼합한 다음, 중화반응액을 희석한 후 적합한 양을 세포에 접종하여 바이러스 함량을 측정한다. (Neutralization reaction) After the reaction of the disinfectant is finished, take out 1.0ml immediately to neutralize the efficacy of the disinfectant, put it in the same amount of neutralization medium of 37, mix, dilute the neutralization reaction solution and inoculate the cells with the appropriate amount of virus content Measure
발육란을 이용한 시험에서는 소독제 중화 반응액 1분량을 5% 비동화 말혈청 199분량과 섞어 소독액을 중화시킨다.In the test using embryonated eggs, disinfectant solution is neutralized by mixing 1 part of disinfectant neutralization reaction solution with 199 parts of 5% unsynthesized horse serum.
(바이러스 증식여부 판정) 반응액 중 바이러스의 증식은 세포를 사용할 경우 단층을 이룬 microplate를 사용하되, 한 소독배수 희석당 4반복 이상으로 접종한 다음, 30분의 흡착을 한 후 배지를 갈아준 후에 37의 CO2 배양기에서 5일간 배양한 후 판독한다. 발육란을 이용한 시험에서는 희석배수당 5개 이상의 발육란을 사용하여 0.2ml를 요막강(allantoic cavity)에 접종하고 적합한 온도에서 배양하되, 매일 관찰한다.(Verification of virus proliferation) Virus propagation in the reaction solution is performed by using monolayered microplate when using cells. After inoculation, after 30 minutes of adsorption, the medium was changed, and then incubated in 37 CO 2 incubator for 5 days, and then read. In the test using embryonated eggs, inoculate 0.2 ml into the allantoic cavity using 5 or more embryonated eggs per dilution and incubate at an appropriate temperature, and observe each day.
(바이러스 함유량 계산) Kaerber method로 한다. (Calculation of Virus Contents) The Kaerber method is used.
(유효배수 평가) 병원체 대조군의 바이러스함량에서 소독제 처리군의 바이러스함량을 대조로 하여 104 PFU(또는 TCID50, EID50, LD50 등)의 불활화가 인정되는 희석농도를 유효농도로 한다.(Effective drainage evaluation) 10 4 PFU (or TCID 50 , EID 50 , LD 50) by comparing the virus content of the disinfectant treatment group with the virus content of the pathogen control group. The dilution concentration at which the inactivation of such a material) is recognized is made an effective concentration.
5. 산성/알칼리제제5. Acid / Alkaline
산성/알칼리제제는 다음과 같은 조건과 방법으로 효력시험과 수소이온농도(pH)를 실시한다. Acid / alkaline preparations are tested for potency and hydrogen ion concentration (pH) under the following conditions and methods.
(소독제 효력시험) 산성/알칼리제제의 효력시험은 본시험 4의 ~항과 동일하게 하되, 유기물희석액은 5% 소태아혈청 대신에 1% 소태아혈청을 사용한다.. (Test of disinfectant efficacy) The efficacy test of acid / alkali preparations should be the same as in ~ 4 of this test, but organic diluent should use 1% fetal bovine serum instead of 5% fetal bovine serum.
(소독제의 수소이온농도 측정) 산성/알칼리제제의 소독제의 수소이온농도(pH)는 이글배지액 (Eagle's medium) 10, 소독제 부유 경수액 40, 소태아혈청 50의 비율로 혼합한 권장희석배수에서 측정한다.
(Measurement of hydrogen ion concentration of disinfectant) The hydrogen ion concentration (pH) of acid / alkali disinfectant is the recommended dilution water mixed with Eagle's medium 10, disinfectant suspended hard water solution 40, and fetal bovine serum 50. Measure
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