KR102475055B1 - Hypochlorous acid drinking composition - Google Patents
Hypochlorous acid drinking composition Download PDFInfo
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- KR102475055B1 KR102475055B1 KR1020200094137A KR20200094137A KR102475055B1 KR 102475055 B1 KR102475055 B1 KR 102475055B1 KR 1020200094137 A KR1020200094137 A KR 1020200094137A KR 20200094137 A KR20200094137 A KR 20200094137A KR 102475055 B1 KR102475055 B1 KR 102475055B1
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- drinking
- water
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- hypochlorite
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Abstract
본 발명은 종래 차아염소산수는 안전성이 부족하여 섭취하기 어렵다는 점을 해결하고자, 유기산 혼합용액 및 차아염소산염 혼합용액을 일정비로 혼합한 음용가능한 음용 조성물에 관한 것이다. 본 발명은 음용 안전성이 있으며, 혈압 강하, 간기능 개선, 총 콜레스테롤 저하 등의 건강 증진 효과를 나타낸다.The present invention relates to a drinkable drinking composition in which an organic acid mixture solution and a hypochlorite mixture solution are mixed at a certain ratio in order to solve the problem that conventional hypochlorous acid water is difficult to ingest due to lack of safety. The present invention is safe for drinking, and exhibits health-promoting effects such as lowering blood pressure, improving liver function, and lowering total cholesterol.
Description
본 발명은 차아염소산 음용 조성물에 관한 것으로, 상세하게는 유기산 혼합용액 및 차아염소산염 혼합용액을 일정비로 혼합하여, 음용 안전성이 있으며, 건강증진효과가 우수한 음용 조성물에 관한 것이다.The present invention relates to a drinking composition for hypochlorous acid, and more particularly, to a drinking composition that is safe for drinking and has excellent health promoting effects by mixing a mixed solution of an organic acid and a mixed solution of hypochlorite at a certain ratio.
일반적으로, 살균력을 나타내는 물질은 살균과 동시에 환경, 인체 등에 악영향을 끼칠 수 있는 가능성이 있다. 살균력을 나타내는 물질 중 염소성분을 포함하는 물질은 세정제, 소독제, 살균제 등으로 널리 사용되고 있고, 이 중 차아염소산 나트륨과 차아염소산 칼슘은 1957년에 살충제로 사용하도록 처음 등록되어 사람과 동물에게 질병을 일으킬 수 있는 박테리아, 곰팡이, 조류 등을 억제하는데 사용되었다. 염소소독제는 염소산 염의 형태일 때 실온에서 안정적이고 유효기간이 긴 장점이 있지만 살균효과가 떨어지고, 염소가스의 휘발로 인한 안전문제가 제기된다(Fisher, 2009). 이산화 염소(chlorin dioxide)는 주로 실내 환경에서 무생물 및 표면의 박테리아, 바이러스, 곰팡이 등 유해한 미생물을 방제하여 식품 소독제, 식품가공장비의 소독, 의료폐기물 처리 등에 사용되는 비교적 인체에 안전한 물질이지만 용액 상태에서 매우 불안정하고 유효기간이 매우 짧아 이용 범위가 한정적이다.In general, a substance exhibiting sterilizing power has the possibility of adversely affecting the environment, human body, etc. at the same time as sterilizing. Among substances showing sterilizing power, substances containing chlorine are widely used as detergents, disinfectants, and sterilizers. It has been used to control bacteria, fungi, and algae that can Chlorine disinfectant has the advantage of being stable at room temperature and having a long shelf life when it is in the form of chlorate salt, but has a low sterilization effect and raises safety issues due to volatilization of chlorine gas (Fisher, 2009). Chlorine dioxide is a relatively safe substance for the human body, mainly used in food disinfectants, disinfection of food processing equipment, medical waste treatment, etc. by controlling harmful microorganisms such as bacteria, viruses, molds, etc. It is very unstable and has a very short shelf life, so the range of use is limited.
안전성과 안정성을 모두 만족하기 위한 대안으로 차아염소산수가 주목받고 있다. 차아염소산수의 주 성분은 차아염소산으로서, 주요 활성 염소종은 차아염소산(Hypochlorous acid, HOCl)과 차아염소산 이온(Hypochlorite ions, OCl-), 유리 염소 및 차아염소산염(Hypochlorite)이며, 그 중에서 차아염소산이 가장 우세한 비중을 차지한다. 차아염소산의 안전성과 안정성의 특성에 의해 미국식품의약국은 1999년부터 강산성 차아염소산수를 농수산물 세정에 사용하도록 허용하였고, 일본후생노동성에서는 2013년 생식용 어류, 생굴, 냉동식품 가공에 첨가물로 허용하고 있으며, 한국의 경우 식품의약품안전청(Korea Food & Drug Administration)의 식품첨가물 기준법에 따라 식품첨가물로 인정하고 있다.Hypochlorous acid water is attracting attention as an alternative to satisfy both safety and stability. The main component of hypochlorous acid water is hypochlorous acid, and the main active chlorine species are hypochlorous acid (HOCl), hypochlorite ions (OCl - ), free chlorine and hypochlorite, among which hypochlorous acid It occupies the most predominant proportion. Due to the safety and stability characteristics of hypochlorous acid, the U.S. Food and Drug Administration has allowed strong acidic hypochlorous acid water to be used for cleaning agricultural and marine products since 1999, and the Ministry of Health, Labor and Welfare of Japan has allowed it as an additive in raw fish, raw oysters, and frozen food processing in 2013. In Korea, it is recognized as a food additive according to the Food Additives Standards Act of the Korea Food & Drug Administration.
다만, 차아염소산수를 식품첨가물로 인정하고 있으나, 안전성을 고려하여 허용 기준치를 설정하고 있다. 미국의 경우, 식품 살충 및 살균제, 야채 및 과일의 세정 시에 200 ppm 미만, 씨앗 살균시 60-80 ppm 이하, 사람 음용 시 4 ppm 이하로 규정하고 있고, 한국의 경우, 차아염소산수는 과일류, 채소류 등의 살균 목적에 한하여 사용하여야하며, 최종식품의 완성 전에 제거하여야 한다고 규정하고 있다.However, although hypochlorous acid water is recognized as a food additive, acceptable standards are set in consideration of safety. In the case of the United States, less than 200 ppm when cleaning food pesticides and fungicides, vegetables and fruits, less than 60-80 ppm when sterilizing seeds, and less than 4 ppm when human drinking, and in Korea, hypochlorous acid water is prescribed for fruits However, it stipulates that it should be used only for the purpose of sterilization of vegetables, etc., and should be removed before final food is completed.
차아염소산수의 살균력, 안정성, 저장성, 친환경성 등의 특성에 의해 활용 분야가 매우 광범위하다. 차아염소산수는 식품가공, 농업, 의료, 수산, 축산 등에서 활용되고 있으며, 의료 분야에서는 손을 포함한 신체 소독, 내시경소독, 화상부위치료, 상처소독 등의 목적으로 이용되고 있다. 이와 관련하여, 대한민국공개특허 제10-2019-0105014호에서는 차아염소산 및 아세트산을 포함하여 피부 외상과 관련된 질환을 치료하기 위한 조성물이 개시되어 있고, 대한민국등록특허 제10-1781229호에서는 차아염소산 및 물을 포함하여, 화상, 절단, 마찰상 등의 조직을 치료하기 위한 약제가 개시되어 있으며, 일본공개특허 제2019-206544호에서는 수술 부위, 창상, 외상 부위에서 염증성 질환을 치료하기 위한 차아염소산염 용액이 개시되어 있다.Hypochlorous acid water has a very wide range of applications due to its sterilizing power, stability, storability, and eco-friendliness. Hypochlorous acid water is used in food processing, agriculture, medicine, fisheries, livestock, etc. In the medical field, it is used for body disinfection including hands, endoscope disinfection, burn treatment, and wound disinfection. In this regard, Korean Patent Publication No. 10-2019-0105014 discloses a composition for treating skin trauma-related diseases including hypochlorous acid and acetic acid, and Korean Patent Registration No. 10-1781229 discloses hypochlorous acid and water Including, drugs for treating tissues such as burns, cuts, and abrasions are disclosed, and in Japanese Patent Publication No. 2019-206544, a hypochlorite solution for treating inflammatory diseases at surgical sites, wounds, and traumatic sites is disclosed. has been initiated.
다만, 차아염소산수의 박테리아, 바이러스, 곰팡이 등에 대한 살균력이 알려져 있더라도, 이는 인체에 들어가는 것을 전제로 하지 않은 것이고, 이를 음용하는 경우 살균력을 나타내는지에 대하여 구체적으로 개시된 바 없으며, 이의 음용시 안정성에 대해서도 구체적으로 개시된 바 없다.However, even if the bactericidal power of hypochlorous acid water against bacteria, viruses, fungi, etc. is known, this is not based on the premise that it enters the human body, and no specific disclosure has been made as to whether it exhibits sterilizing power when drinking it, and also about its stability during drinking. Not specifically disclosed.
이에, 본 발명자들은 희석식으로 제조된 차아염소산 음용 조성물을 제조하기 위해 노력한 결과, 음용하더라도 간기능 지표, 체중 등에 차이가 없음을 확인하여 안정성이 있다는 점 뿐만 아니라 혈압 강하, 총 콜레스테롤 감소 등의 우수한 효과가 있음을 밝힘으로써, 본 발명을 완성하였다.Accordingly, the present inventors have tried to prepare a hypochlorous acid drinking composition prepared by dilution, and as a result, it was confirmed that there is no difference in liver function index, body weight, etc. even after drinking it, and it is stable, as well as excellent effects such as lowering blood pressure and reducing total cholesterol. By revealing that there is, the present invention was completed.
본 발명에서는 종래 차아염소산은 염소농도 등에 의해 안전성이 부족하다는 점을 해결하고자, 음용가능한 구연산, 아세트산 및 개미산을 이용하며, 이를 차아염소산 나트륨 및 규소염과 혼합 및 희석하여 음용가능한 음용 조성물을 제공하기 위한 것이다. 또한, 음용을 통해 혈압 강하 등의 건강 증진이 가능한 음용 조성물을 제공하기 위한 것이다.In the present invention, in order to solve the lack of safety of conventional hypochlorous acid due to chlorine concentration, drinkable citric acid, acetic acid, and formic acid are used, and they are mixed and diluted with sodium hypochlorite and silicon salt to provide a drinkable drinking composition. it is for In addition, it is to provide a drinking composition capable of improving health such as lowering blood pressure through drinking.
상기 목적을 달성하기 위하여, 본 발명은 유기산 혼합용액 및 차아염소산염 혼합용액을 물과 혼합하여 제조되는 음용 조성물을 제공한다.In order to achieve the above object, the present invention provides a drinking composition prepared by mixing an organic acid mixture solution and a hypochlorite mixture solution with water.
본 발명의 일 양태에서, 유기산 혼합용액은 아세트산, 구연산 및 개미산으로 구성된 그룹으로부터 선택된 1종 이상을 포함한다. 구체적인 본 발명의 일 양태에서, 상기 유기산 혼합용액은 유기산 혼합용액 전체 중량대비 물 50 내지 99 중량부 및 유기산 혼합물 1 내지 50 중량부이고, 상기 유기산 혼합물은 유기산 혼합물 전체 중량대비 아세트산 40 내지 89 중량부, 구연산 10 내지 60 중량부 및 개미산 1 내지 20 중량부이다.In one aspect of the present invention, the organic acid mixture contains at least one selected from the group consisting of acetic acid, citric acid and formic acid. In a specific aspect of the present invention, the organic acid mixture is 50 to 99 parts by weight of water and 1 to 50 parts by weight of the organic acid mixture based on the total weight of the organic acid mixture, and the organic acid mixture is 40 to 89 parts by weight of acetic acid based on the total weight of the organic acid mixture , 10 to 60 parts by weight of citric acid and 1 to 20 parts by weight of formic acid.
본 발명의 일 양태에서, 차아염소산염 혼합용액은 차아염소산나트륨 또는 규소염을 포함한다. 구체적인 본 발명의 일 양태에서, 상기 차아염소산염 용액은 차아염소산염 용액 전체 중량대비 물 80 내지 99 중량부 및 차아염소산염 혼합물 1 내지 20 중량부이고, 상기 차아염소산염 혼합물은 차아염소산염 혼합물 전체 중량대비 차아염소산나트륨 70 내지 99 중량부 및 규소염 1 내지 30 중량부이다.In one aspect of the present invention, the hypochlorite mixed solution includes sodium hypochlorite or silicon salt. In a specific aspect of the present invention, the hypochlorite solution is 80 to 99 parts by weight of water and 1 to 20 parts by weight of the hypochlorite mixture based on the total weight of the hypochlorite solution, and the hypochlorite mixture is sodium hypochlorite based on the total weight of the hypochlorite mixture 70 to 99 parts by weight and 1 to 30 parts by weight of a silicon salt.
또한, 본 발명의 일 양태에서, 음용 조성물은 음용 조성물 전체중량대비 유기산 혼합용액 0.01 내지 10 중량부, 차아염소산염 혼합용액 0.01 내지 10 중량부 및 물 80 내지 99.98 중량부이다.Further, in one aspect of the present invention, the drinking composition comprises 0.01 to 10 parts by weight of the organic acid mixture solution, 0.01 to 10 parts by weight of the hypochlorite mixture solution, and 80 to 99.98 parts by weight of water, based on the total weight of the drinking composition.
본 발명의 일 양태에서, 음용 조성물은 염소 부산물을 포함하지 않는다.In one aspect of the invention, the drinkable composition is free of chlorine byproducts.
본 발명의 일 양태에서, 음용 조성물의 유효염소농도가 1 내지 200 ppm이다. 구체적인 본 발명의 일 양태에서, 음용 조성물의 유효염소농도가 4 ppm 초과 150 ppm 미만이다. 보다 구체적인 본 발명의 일 양태에서, 음용 조성물의 유효염소농도가 40 내지 60 ppm이다.In one aspect of the present invention, the effective chlorine concentration of the drinking composition is 1 to 200 ppm. In a specific aspect of the present invention, the effective chlorine concentration of the drinking composition is greater than 4 ppm and less than 150 ppm. In one more specific aspect of the present invention, the effective chlorine concentration of the drinking composition is 40 to 60 ppm.
또한, 본 발명은 간 질환 및 대사성 질환으로 구성된 그룹으로부터 선택된 1종 이상의 질환 예방 또는 개선 효과를 제공한다.In addition, the present invention provides an effect of preventing or improving one or more diseases selected from the group consisting of liver diseases and metabolic diseases.
본 발명의 일 양태에서, 대사성 질환은 비만, 당뇨, 고지혈증, 지방간, 심혈관질환, 갑상선 항진증, 갑상선 저하증, 고혈압, 저혈압, 고혈당, 저혈당 및 통풍으로 이루어진 그룹으로부터 선택되는 것이고, 간 질환은 간염, 간경변, 비알콜성 지방간, 알코올성 지방간, 및 간경변으로 이루어진 군에서 선택되는 것이다.In one aspect of the present invention, the metabolic disease is selected from the group consisting of obesity, diabetes, hyperlipidemia, fatty liver, cardiovascular disease, hyperthyroidism, hypothyroidism, hypertension, hypotension, hyperglycemia, hypoglycemia and gout, and the liver disease is hepatitis, cirrhosis , It is selected from the group consisting of non-alcoholic fatty liver, alcoholic fatty liver, and liver cirrhosis.
본 발명의 일 양태에서, 음용 조성물은 혈압강하, 혈당감소, 총 콜레스테롤 감소, 저밀도 콜레스테롤 감소, AST 활성감소, ALT 활성감소 및 감마지티피(γ-GTP) 활성감소로 구성된 그룹으로부터 하나 이상의 효과를 나타낸다.In one aspect of the present invention, the drinking composition has at least one effect from the group consisting of lowering blood pressure, lowering blood sugar, lowering total cholesterol, lowering low-density cholesterol, reducing AST activity, reducing ALT activity, and reducing gamma-GTP activity. indicate
본 발명은 유기산 혼합용액 및 차아염소산염 혼합용액을 일정비로 혼합한 음용 조성물에 관한 것으로, 음용 안전성이 있으며, 음용시 혈압 강하, 간기능 개선, 총 콜레스테롤 저하 등의 건강 증진 효과가 우수하다는 이점이 있다.The present invention relates to a drinking composition in which an organic acid mixture solution and a hypochlorite mixture solution are mixed at a certain ratio, and has the advantage of being safe for drinking and having excellent health promotion effects such as lowering blood pressure, improving liver function, and lowering total cholesterol when drinking. .
도 1a은 마우스의 체중 무게 변화를 나타낸 도이다.
도 1b는 마우스의 간 무게 변화를 나타낸 도이다.
도 2a는 혈청과 간 균질액에서 총 활성산소의 양을 나타낸 도이다.
도 2b는 혈청과 간 균질액에서 산화질소의 양을 나타낸 도이다.
도 3a는 혈청과 간 균질액에서 항산화효소 GPx의 농도를 나타낸 도이다.
도 3b는 혈청과 간 균질액에서 항산화효소 카탈라제의 농도를 나타낸 도이다.
도 4a는 혈청과 간 균질액에서 사이토카인 IL-1β의 양을 나타낸 도이다.
도 4b는 혈청과 간 균질액에서 사이토카인 IL-12을 나타낸 도이다.
도 4c는 혈청과 간 균질액에서 사이토카인 IFN-γ을 나타낸 도이다.
도 4d는 혈청과 간 균질액에서 사이토카인 TNF-α을 나타낸 도이다.
도 4e는 혈청과 간 균질액에서 사이토카인 IL-6을 나타낸 도이다.
도 4f는 혈청과 간 균질액에서 사이토카인 IL-10을 나타낸 도이다.
도 5는 간에서 총 ALT 농도 및 AST 농도를 나타낸 도이다.
도 6은 간의 조직학적 변화를 나타낸 도이다.Figure 1a is a diagram showing the change in body weight of the mouse.
Figure 1b is a diagram showing the change in liver weight of mice.
Figure 2a is a diagram showing the amount of total active oxygen in serum and liver homogenate.
Figure 2b is a diagram showing the amount of nitric oxide in serum and liver homogenate.
Figure 3a is a diagram showing the concentration of antioxidant enzyme GPx in serum and liver homogenate.
Figure 3b is a diagram showing the concentration of the antioxidant enzyme catalase in serum and liver homogenate.
Figure 4a is a diagram showing the amount of the cytokine IL-1β in serum and liver homogenate.
Figure 4b is a diagram showing the cytokine IL-12 in serum and liver homogenate.
Figure 4c is a diagram showing the cytokine IFN-γ in serum and liver homogenate.
Figure 4d is a diagram showing the cytokine TNF-α in serum and liver homogenate.
Figure 4e is a diagram showing the cytokine IL-6 in serum and liver homogenate.
Figure 4f is a diagram showing the cytokine IL-10 in serum and liver homogenate.
5 is a diagram showing total ALT and AST concentrations in the liver.
6 is a diagram showing histological changes in the liver.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 명세서 전체에서, 어떤 부분이 어떤 구성 요소를 “포함”한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성 요소를 제외하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다.Throughout the specification of the present invention, when a certain component is said to "include", it means that it may further include other components without excluding other components unless otherwise stated.
본 명세서에서 "약학적으로 허용가능한" 및 "식품학적으로 허용가능한"이란 포함된 성분이 생물체를 상당히 자극하지 않고, 생물학적 활성 및 특성을 저해하지 않는 것을 의미한다.As used herein, "pharmaceutically acceptable" and "food pharmaceutically acceptable" mean that the contained components do not significantly stimulate living organisms and do not interfere with biological activities and properties.
본 명세서에서 기재되는 화합물은 특별히 정의를 하지 않는 한, 통상적으로 인식된 화합물을 의미한다.A compound described herein means a commonly recognized compound unless otherwise defined.
본 발명에서 사용되는 용어 "예방"은 본 발명의 조성물의 체내 투입으로 특정 질환(예를 들어, 고혈압)의 증상을 억제시키거나 진행을 지연시키는 모든 행위를 의미한다.The term "prevention" used in the present invention refers to any action that suppresses symptoms or delays the progression of a specific disease (eg, high blood pressure) by injecting the composition of the present invention into the body.
본 발명에서 사용되는 용어 "개선"은 본 발명의 조성물의 체내 투입으로 특정 질환(예를 들어, 고혈압)의 증상과 관련된 파라미터, 예를 들면 혈압 강하 등 증상의 정도를 감소시키는 모든 행위를 의미한다.The term "improvement" used in the present invention refers to any activity that reduces the degree of a parameter related to symptoms of a specific disease (eg, high blood pressure), such as lowering blood pressure, by injecting the composition of the present invention into the body. .
본 발명의 “혈압 강하” 는 혈압이 정상 혈압범위(80mmHg 내지 120mmHg)를 초과하는 대상에 대하여 혈압을 정상 혈압 범위로 낮춘다는 의미로 사용된다."Blood pressure lowering" of the present invention is used in the sense of lowering the blood pressure to the normal blood pressure range for subjects whose blood pressure exceeds the normal blood pressure range (80 mmHg to 120 mmHg).
본 발명은 유기산 혼합용액 및 차아염소산염 혼합용액을 물과 혼합하여 제조되는 음용 조성물에 관한 것이다.The present invention relates to a drinking composition prepared by mixing an organic acid mixture solution and a hypochlorite mixture solution with water.
또한, 본 발명은 상기 음용 조성물을 통한 간 질환 및 대사성 질환으로 구성된 그룹으로부터 선택된 1종 이상의 질환 예방 또는 개선 효과에 관한 것이다. 또한, 본 발명 음용 조성물은 구강 치아 및 위장관 조직에 영향을 주지않아, 음용 시 구강 살균도 가능하다.In addition, the present invention relates to an effect of preventing or improving one or more diseases selected from the group consisting of liver disease and metabolic disease through the drinking composition. In addition, the drinking composition of the present invention does not affect oral teeth and gastrointestinal tissues, and oral sterilization is also possible during drinking.
본 발명의 일 양태에서, 유기산 혼합용액은 아세트산(acetic acid), 구연산(citric acid) 및 개미산(formic acid)으로 구성된 그룹으로부터 선택된 1종 이상을 포함하는 것이다. 구체적인 본 발명의 일 양태에서, 상기 유기산 혼합용액은 유기산 혼합용액 전체 중량대비 물 50 내지 99 중량부 및 유기산 혼합물 1 내지 50 중량부이고, 상기 유기산 혼합물은 유기산 혼합물 전체 중량대비 아세트산 40 내지 89 중량부, 구연산 10 내지 60 중량부 및 개미산 1 내지 20 중량부이다. 보다 더 구체적인 본 발명의 일 양태에서, 상기 유기산 혼합용액은 유기산 혼합용액은 유기산 혼합용액 전체 중량대비 물 90 내지 99 중량부 및 유기산 혼합물 1 내지 10 중량부이고, 상기 유기산 혼합물 전체 중량대비 아세트산 40 내지 89 중량부, 구연산 10 내지 60 중량부 및 개미산 1 내지 20 중량부이다.In one aspect of the present invention, the organic acid mixed solution is one containing at least one selected from the group consisting of acetic acid, citric acid, and formic acid. In a specific aspect of the present invention, the organic acid mixture is 50 to 99 parts by weight of water and 1 to 50 parts by weight of the organic acid mixture based on the total weight of the organic acid mixture, and the organic acid mixture is 40 to 89 parts by weight of acetic acid based on the total weight of the organic acid mixture , 10 to 60 parts by weight of citric acid and 1 to 20 parts by weight of formic acid. In a more specific aspect of the present invention, the organic acid mixture solution is 90 to 99 parts by weight of water and 1 to 10 parts by weight of the organic acid mixture based on the total weight of the organic acid mixture, and 40 to 40 parts by weight of acetic acid based on the total weight of the organic acid mixture 89 parts by weight, 10 to 60 parts by weight of citric acid and 1 to 20 parts by weight of formic acid.
본 발명에서 유기산 혼합용액은 아세트산, 구연산 및 개미산 이외에도 유기산으로 알려진 화합물을 더 포함할 수 있고, 예를 들면, 인산(phosphoric acid), 푸마르산(fumaric acid), 젖산(lactic acid), 사과산(malic acid) 등을 더 포함할 수 있다.In the present invention, the organic acid mixed solution may further contain a compound known as an organic acid in addition to acetic acid, citric acid and formic acid, for example, phosphoric acid, fumaric acid, lactic acid, malic acid ) and the like may be further included.
본 발명의 일 양태에서, 차아염소산염 혼합용액은 차아염소산 나트륨 또는 규소염을 포함하는 것이다. 구체적인 본 발명의 일 양태에서, 상기 차아염소산염 용액은 차아염소산염 용액 전체 중량대비 물 80 내지 99 중량부 및 차아염소산염 혼합물 1 내지 20 중량부이고, 상기 차아염소산염 혼합물은 차아염소산염 혼합물 전체 중량대비 차아염소산나트륨 70 내지 99 중량부 및 규소염 1 내지 30 중량부이다.In one aspect of the present invention, the hypochlorite mixed solution contains sodium hypochlorite or silicon salt. In a specific aspect of the present invention, the hypochlorite solution is 80 to 99 parts by weight of water and 1 to 20 parts by weight of the hypochlorite mixture based on the total weight of the hypochlorite solution, and the hypochlorite mixture is sodium hypochlorite based on the total weight of the hypochlorite mixture 70 to 99 parts by weight and 1 to 30 parts by weight of a silicon salt.
본 발명의 일 양태에서, 음용 조성물은 음용 조성물 전체중량대비 유기산 혼합용액 0.01 내지 10 중량부, 차아염소산염 혼합용액 0.01 내지 10 중량부이고, 나머지는 물에 해당한다. 구체적인 본 발명의 일 양태에서, 음용 조성물은 전체중량대비 유기산 혼합용액 0.01 내지 5 중량부, 차아염소산염 혼합용액 0.01 내지 5 중량부이고, 나머지는 물에 해당한다.In one aspect of the present invention, the drinking composition comprises 0.01 to 10 parts by weight of the organic acid mixture solution and 0.01 to 10 parts by weight of the hypochlorite mixture solution, with the remainder corresponding to water, based on the total weight of the drinking composition. In a specific aspect of the present invention, the composition for drinking is 0.01 to 5 parts by weight of the organic acid mixture solution and 0.01 to 5 parts by weight of the hypochlorite mixture solution based on the total weight, and the remainder corresponds to water.
본 발명에서 구연산, 아세트산, 개미산, 차아염소산 나트륨 및 규소염은 상업적으로 판매되는 것을 사용하거나, 당업계에 공지된 방법으로 합성하거나, 자연에서 채취한 후 처리하여 수득된 것을 사용할 수 있으며, 이에 한정되는 것은 아니다. 또한, 규소염은 규소가 염의 형태를 나타내는 것으로, 이에 한정되는 것은 아니나 예를 들면, [SiO3]2-, [HSiO3]- 등을 포함한다.Citric acid, acetic acid, formic acid, sodium hypochlorite, and silicon salt in the present invention may be used commercially available, synthesized by a method known in the art, or obtained by processing after being collected from nature, limited thereto it is not going to be In addition, the silicon salt represents a salt form of silicon, but is not limited thereto, and includes, for example, [SiO 3 ] 2- , [HSiO 3 ] -, and the like.
본 발명에서 차아염소산 나트륨을 포함한 차아염소산 음용 조성물의 제조는 구연산, 아세트산, 개미산, 차아염소산 나트륨 및 규소염을 포함하는 혼합물은 물과 혼합하여 희석하는 방법에 의해 제조될 수 있다. 보다 상세하게는 구연산, 아세트산 및 개미산을 혼합한 혼합용액을 제조한 후, 차아염소산 나트륨 및 규소염을 혼합하여 혼합물을 제조하고, 물과 혼합하여 희석시켜 음용 조성물을 제조할 수 있다. 혼합되는 순서는 구연산-아세트산-개미산-차아염소산 나트륨-규소염-물의 순서로 혼합될 수 있으나, 물에 구연산, 아세트산, 개미산, 차아염소산 나트륨 및 규소염을 투입하는 방식, 차아염소산 나트륨을 물과 혼합한 뒤 구연산, 아세트산 및 개미산의 혼합용액을 혼합하는 방식 등 순서에 무관하게 혼합하여 제조할 수 있다.In the present invention, the preparation of the hypochlorite drinking composition containing sodium hypochlorite can be prepared by mixing and diluting a mixture containing citric acid, acetic acid, formic acid, sodium hypochlorite and silicon salt with water. More specifically, after preparing a mixed solution in which citric acid, acetic acid and formic acid are mixed, a mixture is prepared by mixing sodium hypochlorite and silicon salt, and mixed with water to dilute to prepare a drinking composition. The order of mixing can be mixed in the order of citric acid-acetic acid-formic acid-sodium hypochlorite-siliconate-water, but a method of adding citric acid, acetic acid, formic acid, sodium hypochlorite and silicon salt to water, sodium hypochlorite with water After mixing, it can be prepared by mixing in any order, such as mixing a mixed solution of citric acid, acetic acid and formic acid.
또한, 본 발명에서 차아염소산 음용 조성물은 희석하는 방식 이외에 전기분해에 의한 방식에 의해서도 제조될 수 있으나, 희석에 의해 제조하는 것이 바람직하다. 희석에 의해 제조되는 경우 음용활용성이 높고, 고농도로 대량 생산이 가능하고, 전기분해 또는 염산을 이용하지 않음으로써 안전하고, 유해성 부산물이 발생하지 않을 수 있다.In addition, in the present invention, the hypochlorous acid drinking composition may be prepared by electrolysis in addition to dilution, but it is preferable to prepare by dilution. When produced by dilution, it is highly drinkable, can be mass-produced at high concentration, is safe by not using electrolysis or hydrochloric acid, and may not generate harmful by-products.
본 발명에서 음용 조성물은 구연산, 아세트산, 개미산, 차아염소산 나트륨 및 규소염을 포함하는 혼합물 이외에 약학적으로 허용가능한 성분 또는 식품학적으로 허용가능한 성분, 예를 들면, 비타민 A, 미네랄, 무기질 등을 더 포함할 수 있다.In the present invention, the drinking composition further contains pharmaceutically acceptable ingredients or food-acceptable ingredients such as vitamin A, minerals, minerals, etc. can include
상기 구연산, 아세트산, 개미산, 차아염소산 나트륨은 유효염소농도를 변경시키기 위해 조절될 수 있다. 또한, 구연산, 아세트산 및 개미산을 이용함으로써 음용시 생체 내 생물학적 활성 또는 특성을 변경시키지 않고, 유의미한 예방, 개선 효과를 나타낼 수 있다.The citric acid, acetic acid, formic acid, and sodium hypochlorite may be adjusted to change the effective chlorine concentration. In addition, by using citric acid, acetic acid, and formic acid, it is possible to exhibit significant preventive and improving effects without altering in vivo biological activity or characteristics during drinking.
본 발명의 일 양태에서, 음용 조성물은 염소 부산물을 포함하지 않는다. 종래 전기분해방식을 이용하지 않음으로써, 염소를 포함하는 발암물질인 염소부산물이 발생하지 않는다.In one aspect of the invention, the drinkable composition is free of chlorine byproducts. By not using the conventional electrolysis method, chlorine by-products, which are carcinogens including chlorine, are not generated.
본 발명의 일 양태에서, 음용 조성물은 유효염소농도가 1 내지 200 ppm이다. In one aspect of the present invention, the drinking composition has an effective chlorine concentration of 1 to 200 ppm.
구체적인 본 발명의 일 양태에서, 음용 조성물은 유효염소농도가 4 ppm 초과 150 ppm 미만이다. 보다 더 구체적인 본 발명의 일 양태에서, 유효염소농도가 10 내지 125 ppm, 25 내지 110 ppm, 40 내지 60 ppm이다. 본 발명에서 음용 조성물은 유효염소농도가 적정수준으로 조절되어야 할 필요가 있으며, 유기산 혼합용액 및 차아염소산염 혼합용액을 통해 유효염소농도가 상기 범위를 초과하는 경우, 예컨대 200 ppm을 초과하는 음용 조성물을 음용하는 경우, 생물학적 활성 또는 특성에 부정적인 영향을 끼칠 수 있다. 한편, 본 발명에서 음용 조성물은 4 ppm의 농도를 초과하더라도 구연산, 아세트산을 포함하고, 희석하는 방식에 의해 제조됨으로써 음용 안전성을 가진다.In one specific aspect of the present invention, the drinking composition has an effective chlorine concentration of greater than 4 ppm and less than 150 ppm. In one more specific aspect of the present invention, the effective chlorine concentration is 10 to 125 ppm, 25 to 110 ppm, 40 to 60 ppm. In the present invention, the drinking composition needs to have the effective chlorine concentration adjusted to an appropriate level, and when the effective chlorine concentration exceeds the above range through the organic acid mixed solution and the hypochlorite mixed solution, for example, a drinking composition exceeding 200 ppm Ingestion may adversely affect biological activity or properties. On the other hand, in the present invention, the drinking composition contains citric acid and acetic acid, even if the concentration exceeds 4 ppm, and is prepared by dilution, so that it has drinking safety.
또한, 본 발명은 상기 음용 조성물을 통해 간 질환 및 대사성 질환으로 구성된 그룹으로부터 선택된 1종 이상의 질환 예방 또는 개선 효과를 제공한다.In addition, the present invention provides an effect of preventing or improving one or more diseases selected from the group consisting of liver disease and metabolic disease through the drinking composition.
상기 대사성 질환은 비만, 당뇨, 고지혈증, 지방간, 심혈관질환, 갑상선 항진증, 갑상선 저하증, 고혈압, 저혈압, 고혈당, 저혈당 및 통풍으로 이루어진 그룹에서 선택되는 어느 하나 이상의 대사성 질환일 수 있다.The metabolic disease may be one or more metabolic diseases selected from the group consisting of obesity, diabetes, hyperlipidemia, fatty liver, cardiovascular disease, hyperthyroidism, hypothyroidism, hypertension, hypotension, hyperglycemia, hypoglycemia, and gout.
또한, 상기 간 질환은 간염, 간경변, 비알콜성 지방간, 알코올성 지방간, 및 간경변으로 이루어진 군에서 선택되는 어느 하나 이상의 간질환일 수 있다. In addition, the liver disease may be any one or more liver diseases selected from the group consisting of hepatitis, cirrhosis, non-alcoholic fatty liver, alcoholic fatty liver, and cirrhosis.
또한, 본 발명의 일 양태에서, 음용 조성물은 혈압강하, 혈당감소, 총 콜레스테롤 감소, 저밀도 콜레스테롤 감소, AST 활성감소, ALT 활성감소 및 감마지티피(γ-GTP) 활성감소로 구성된 그룹으로부터 하나 이상의 효과를 나타낸다.In addition, in one aspect of the present invention, the drinking composition is at least one from the group consisting of lowering blood pressure, reducing blood sugar, reducing total cholesterol, reducing low-density cholesterol, reducing AST activity, reducing ALT activity, and reducing gamma-GTP activity. show effect.
구체적으로, 본 발명의 조성물을 음용하여 대사성 질환의 일종인 고혈압, 고지혈증, 심혈관질환 등과 관련된 주요 지표인, 혈압, 총 콜레스테롤, 고밀도 콜레스테롤, 저밀도 콜레스테롤의 수치를 정상 수치까지 개선시킬 수 있다.Specifically, by drinking the composition of the present invention, the levels of blood pressure, total cholesterol, high-density cholesterol, and low-density cholesterol, which are major indicators related to hypertension, hyperlipidemia, and cardiovascular disease, which are types of metabolic diseases, can be improved to normal levels.
또한, 본 발명의 조성물을 음용하여 간염, 간경변, 지방간 및 간경화과 관계된 간손상의 주요 지표인, 간 효소 ALT(Alanine transaminase) 및 AST(Aspartate transaminase), 감마지티피(γ-GTP)의 농도를 정상 수치까지 감소시킬 수 있다.In addition, by drinking the composition of the present invention, the concentration of liver enzymes ALT (Alanine transaminase) and AST (Aspartate transaminase), and gamma-GTP (γ-GTP), which are major indicators of liver damage related to hepatitis, cirrhosis, fatty liver and cirrhosis, are normalized. number can be reduced.
또한, 본 발명의 조성물을 음용하더라도 세포 독성과 관련된 활성산소(reactive oxygen species, ROS), 산화질소(nitric oxide, NO), 항산화효소 GPx(Glutathione Peroxidase), 항산화효소 카탈라제(catalase), 혈청 사이토카인(cytokine)의 수치에 유의한 차이를 나타내지 않거나 오히려 개선된 효과를 나타내므로, 음용 조성물로서 적합하다. In addition, even if the composition of the present invention is drunk, reactive oxygen species (ROS), nitric oxide (NO), antioxidant enzyme GPx (glutathione peroxidase), antioxidant enzyme catalase, and serum cytokines related to cytotoxicity Since it does not show a significant difference in the level of cytokine or rather shows an improved effect, it is suitable as a drinking composition.
이하, 본 발명을 실시예 및 실험예에 의해 상세히 설명한다.Hereinafter, the present invention will be described in detail by examples and experimental examples.
단, 하기 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예 및 실험예에 한정되는 것은 아니다.However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following Examples and Experimental Examples.
<실시예> 차아염소산 음용수 제조<Example> Production of hypochlorous acid drinking water
<1-1> 유기산 혼합용액 제조<1-1> Preparation of organic acid mixture solution
아세트산, 구연산 및 개미산을 혼합하고, 물을 추가로 혼합하여 유기산 혼합용액을 제조하였다. 유기산 혼합용액은 아세트산 15 mg, 구연산 6.25 mg 및 개미산 3.75 mg을 혼합하고, 물을 480 ml 추가로 혼합하여 제조되었다.Acetic acid, citric acid, and formic acid were mixed, and water was further mixed to prepare an organic acid mixture solution. A mixed organic acid solution was prepared by mixing 15 mg of acetic acid, 6.25 mg of citric acid and 3.75 mg of formic acid, and further mixing with 480 ml of water.
<1-2> 차아염소산염 혼합용액 제조<1-2> Preparation of hypochlorite mixed solution
차아염소산나트륨 및 규소염을 혼합하고, 물을 추가로 혼합하여 차아염소산염 혼합용액을 제조하였다. 차아염소산염 혼합용액은 차아염소산나트륨 40 mg 및 규소염 10 mg을 물을 450 ml와 혼합하여 제조되었다.A mixed solution of hypochlorite was prepared by mixing sodium hypochlorite and silicon salt and further mixing with water. A mixed solution of hypochlorite was prepared by mixing 40 mg of sodium hypochlorite and 10 mg of silicon salt with 450 ml of water.
<1-3> 차아염소산 음용수 제조<1-3> Production of hypochlorous acid drinking water
상기 실시예 <1-1>에서 제조된 유기산 혼합용액 및 <1-2>에서 제조된 차아염소산염 혼합용액을 혼합하여 차아염소산 조성물을 제조하였다. 차아염소산 조성물은 유기산 혼합용액 30 ml 및 차아염소산염 혼합용액 100 ml을 혼합하여 제조되었고, 차아염소산 조성물을 물 1~50 L에 희석하여 저농도 차아염소산 음용수(MS-HOCLL), 중농도 차아염소산 음용수(MS-HOCLM), 고농도 차아염소산 음용수(MS-HOCLH)를 제조하였으며, 그 특성은 표 1에 나타난 바와 같다.A hypochlorous acid composition was prepared by mixing the organic acid mixture solution prepared in Example <1-1> and the hypochlorite mixture solution prepared in <1-2>. The hypochlorous acid composition was prepared by mixing 30 ml of the organic acid mixture solution and 100 ml of the hypochlorite mixture solution, and the hypochlorous acid composition was diluted in 1 to 50 L of water to obtain low-concentration hypochlorous acid drinking water (MS-HOCL L ) and medium-concentration hypochlorous acid drinking water. (MS-HOCL M ) and high-concentration hypochlorous acid drinking water (MS-HOCL H ) were prepared, the characteristics of which are shown in Table 1.
<비교예> 미산성전해수 제조<Comparative Example> Production of non-acidic electrolytic water
전기분해장치(HOCLER Cosmic Round Korea Co.,Ltd.)를 사용하여 무격막시스템의 전해조에서 4.5 % 염산이 혼합된 수돗물을 전기분해하여 미산성전해수(CW)를 제조하였다. 미산성전해수의 조성은 표 1에 나타난 바와 같다.By using an electrolysis device (HOCLER Cosmic Round Korea Co., Ltd.), tap water mixed with 4.5% hydrochloric acid was electrolyzed in a non-diaphragm system electrolytic cell to produce slightly acidic electrolyzed water (CW). The composition of non-acidic electrolytic water is shown in Table 1.
(* TW : 수돗물 / CW : 미산성전해수 / ORP : 산화환원 전위차 / ACC : 유효염소농도)(* TW: Tap water / CW: Slightly acidic electrolytic water / ORP: Oxidation-reduction potential difference / ACC: Effective chlorine concentration)
<실험예 1> 실험조건 및 측정방법<Experimental Example 1> Experimental conditions and measurement method
<1-1> 실험용 마우스의 실험조건<1-1> Experimental Conditions for Experimental Mice
6주령(19-20g)의 C57BL6 암컷 마우스 60마리(주식회사 오리엔트)를 구입하여 실험에 사용하였다. 마우스는 온도 23±2°C, 습도 55±5%, 12 h light/dark cycle 환경에서 관리되었다. 일주일 동안 순화과정을 거친 후, 임의로 10마리씩 하기 5개 군으로 분류하였다:Sixty 6-week-old (19-20 g) C57BL6 female mice (Orient Co., Ltd.) were purchased and used in the experiment. Mice were managed in an environment with a temperature of 23±2°C, humidity of 55±5%, and a 12 h light/dark cycle. After a week of acclimatization, they were randomly divided into 5 groups of 10 animals each:
i) 수돗물 음용 대조군(TW), ii) 미산성 전기분해수 음용군(CW), iii) 저농도 MS-차아염소산수(MS-HOClL), iv) 중농도 저농도 MS-차아염소산수(MS-HOClM), v) 고농도 저농도 MS-차아염소산수(MS-HOClH). i) tap water drinking control group (TW), ii) non-acidic electrolyzed water drinking group (CW), iii) low-concentration MS-hypochlorous acid water (MS-HOCl L ), iv) medium-concentration low-concentration MS-hypochlorous acid water (MS-HOCl L ) HOCl M ), v) high concentration low concentration MS-hypochlorous acid water (MS-HOCl H ).
실험수는 마우스용 물병을 통해 자유롭게 음용하게 하였으며 24시간 마다 실험수를 교체하여 주었고 음용기간은 3개월이었다. 실험 동물의 사용 및 관리는 연세대학교 원주의과대학 동물윤리위원회의 승인을 얻은 후 U.S. National Institutes of Health (NIH)의 가이드라인을 따라 수행되었다(YWC-190410-1).The experimental water was allowed to drink freely through a water bottle for mice, and the experimental water was replaced every 24 hours, and the drinking period was 3 months. The use and management of laboratory animals was approved by the Animal Ethics Committee of Wonju College of Medicine, Yonsei University, and the U.S. It was performed according to the guidelines of the National Institutes of Health (NIH) (YWC-190410-1).
<1-2> 체중과 간의 무게 측정<1-2> Weight measurement of body weight and liver
마우스의 체중은 전자저울을 이용하여 주 일 회 12주 동안 측정하였고 체중 차이에 따른 퍼센트로 계산되었다. 간의 무게 또한 도살 직후 분리되어 전자저울을 이용하여 측정되었다. The body weight of the mice was measured once a week for 12 weeks using an electronic balance, and the weight difference was calculated as a percentage. Livers were also weighed immediately after slaughter and measured using an electronic balance.
<1-3> 혈청과 조직샘플의 준비<1-3> Preparation of serum and tissue samples
마우스 혈청과 조직샘플의 준비는 이소플루란(isoflurane, 하나제약)으로 마취시킨 후 표준 프로토콜에 의해 진행되었다. Preparation of mouse serum and tissue samples was performed according to standard protocols after anesthesia with isoflurane (Hana Pharmaceutical).
혈액은 안와정맥총에서 채혈되어 실온에서 응고시킨 후 14,000rpm으로 5분 동안 원심분리하여 혈청을 얻었다. Blood was collected from the orbital venous plexus, coagulated at room temperature, and then centrifuged at 14,000 rpm for 5 minutes to obtain serum.
간 조직은 도살 직후 같은 부위의 조직을 3x3x3mm 크기로 절제한 후 프로테아제 억제제(protease inhibitor)가 혼합된 10 % 용해 완충액(10% cold RIPA lysis buffer, Thermo Scientific)에 넣고 비드 밀링 방법(bead milling method, QIAGEN)을 이용하여 균질화시켰다. 균질액을 4 ℃, 10,000 x g로 15분 동안 원심분리하여 상층액을 분리한 후, 분석 전까지 -80 ℃ 냉동실에 보관하였다. For liver tissue, immediately after slaughter, tissue from the same area was resected to a size of 3x3x3mm, put in 10% lysis buffer (10% cold RIPA lysis buffer, Thermo Scientific) mixed with protease inhibitors, and bead milling method (bead milling method, QIAGEN) was used to homogenize. The homogenate was centrifuged at 4 °C and 10,000 x g for 15 minutes to separate the supernatant, and stored in a -80 °C freezer until analysis.
조직학적 관찰을 위해 약 5x5mm 크기로 간의 같은 부위를 각각 절제한 후 10% 포르말린 용액에 고정하였고, 수세를 통한 고정액 제거, 알코올 탈수, 자일렌(xylene) 치환, 파라핀 침투 과정을 거쳐 파라핀 블록을 만든 후 4 μm 두께로 절편을 만들었다. 절편은 통상적인 방법에 따라 탈파라핀, 함수, 헤마톡실린-에오신(hematoxylin-esoin) 염색, 탈수, 투명, 봉입 과정을 거쳐 광학현미경(BX51, Olympus Co.) 하에서 관찰되었다.For histological observation, the same part of the liver was resected to a size of about 5x5 mm, fixed in 10% formalin solution, and paraffin blocks were made through the process of removing the fixative through washing, alcohol dehydration, xylene substitution, and paraffin infiltration. Sections were then made at 4 μm thickness. Sections were observed under an optical microscope (BX51, Olympus Co.) after deparaffinization, hydrolysis, hematoxylin-esoin staining, dehydration, transparency, and encapsulation according to conventional methods.
<1-4> 산화 스트레스 마커<1-4> Oxidative stress markers
① 혈청과 간조직 내 활성산소(reactive oxygen species, ROS) 농도 측정① Measurement of reactive oxygen species (ROS) concentration in serum and liver tissue
혈청과 간조직에서의 활성산소 농도는 2’,7’-디클로로플루오레신디아세테이트 분석 키트(2’,7’-dichlorofluorescindiacetate(DCFH-DA) assay kit, Abcam)를 이용하여 정해진 프로토콜에 따라 측정되었다. 각 실험군의 샘플들은 10μmol/L의 DCFH-DA와 37 ℃의 암실의 조건에서 30분 동안 반응한 후, 마이크로 플레이트 리더(DTX-880 multimode microplate reader, Beckman Counter Inc.)를 이용하여 488/525 nm에서 흡광도를 측정하였다.The concentration of active oxygen in serum and liver tissue was measured according to a defined protocol using a 2',7'-dichlorofluorescindiacetate (DCFH-DA) assay kit (Abcam). . Samples from each experimental group were reacted with 10 μmol/L of DCFH-DA in a dark room at 37 °C for 30 minutes, and then 488/525 nm using a microplate reader (DTX-880 multimode microplate reader, Beckman Counter Inc.) The absorbance was measured at.
② 혈청과 간조직에서 산화질소(nitric oxide, NO) 농도 측정 ② Measurement of nitric oxide (NO) concentration in serum and liver tissue
산화질소 농도는 NO 탐지 키트(NO detection kit, Intron Biotechnology Company)를 이용하여 정해진 프로토콜에 따라 측정되었다. 50 μL의 혈청과 간 균질액을 각각 96-well plate에 넣고 A시약과 10분, B시약과 10분 동안 실온에서 반응시켰다. 흡광도는 마이크로 플레이트 리더(DTX-880 multimode microplate reader, Beckman Counter Inc.)을 이용하여 540nm에서 측정되었다.The nitric oxide concentration was measured according to a defined protocol using a NO detection kit (Intron Biotechnology Company). 50 μL of serum and liver homogenate were each put into a 96-well plate and reacted with reagent A for 10 minutes and reagent B for 10 minutes at room temperature. Absorbance was measured at 540 nm using a microplate reader (DTX-880 multimode microplate reader, Beckman Counter Inc.).
③ 항산화효소 Glutathione Peroxidase (GPx) 활성 측정③ Antioxidant enzyme Glutathione Peroxidase (GPx) activity measurement
혈청과 간조직 내에서의 GPx 활성은 GPx 분석 키트(GPx assay kit, Biovision)를 이용하여 과산화수소를 제거하는 능력을 통해 평가되었다. GPx에 의해 과산화수소가 환원되면 산화 글루타티온(oxidized glutathione)이 생성되고, 이 물질은 글루타티온 환원 효소(glutathione reductase)와 환원된 니코틴아미드 아데닌 디뉴클레오티드 포스페이트(reduced nicotinamide adenine dinucleotide phosphate(NADPH))에 의해 환원되는 싸이클을 반복한다. NADPH가 NADP+로 산화되는 정도가 될 때 되는 것은 흡광도의 감소로 나타나고, 마이크로 플레이트 리더(DTX-880 multimode microplate reader, Beckman Counter Inc.)를 이용하여 340nm에서 흡광도가 측정되었다.GPx activity in serum and liver tissue was evaluated by the ability to remove hydrogen peroxide using a GPx assay kit (Biovision). When hydrogen peroxide is reduced by GPx, oxidized glutathione is produced, which is reduced by glutathione reductase and reduced nicotinamide adenine dinucleotide phosphate (NADPH). repeat the cycle What happens when NADPH is oxidized to NADP + is indicated by a decrease in absorbance, and absorbance is measured at 340 nm using a microplate reader (DTX-880 multimode microplate reader, Beckman Counter Inc.).
④ 항산화효소 카탈라제(Catalase) 활성 측정④ Measurement of antioxidant enzyme catalase activity
혈청과 간조직 내에서 카탈라제 활성은 카탈라제 색도계 분석 키트(catalase colorimetric assay kit, BioVision)를 이용하여 측정되었다. 과산화수소의 해리율은 스펙트로메터(Beckman Counter Inc.) 570nm에서 측정되었다. 카탈라제 1 unit은 1분 안에 과산화수소 1μM을 분해하는데 필요한 효소의 양으로 정의된다. 카탈라제 활성은 측정법에서 사용된 단백질의 mg으로 노말라이즈(normalize)하였고, mU/mg으로 표현되었다.Catalase activity in serum and liver tissue was measured using a catalase colorimetric assay kit (BioVision). The dissociation rate of hydrogen peroxide was measured at 570 nm with a spectrometer (Beckman Counter Inc.). One unit of catalase is defined as the amount of enzyme required to degrade 1 μM of hydrogen peroxide in 1 minute. Catalase activity was normalized to mg of protein used in the assay and expressed as mU/mg.
⑤ 사이토카인 측정⑤ Cytokine measurement
인터루킨(Interleukin(IL))-1β, IL-6, IL-10, IL-12, TNF-α(tumor necrosis alpha), 인터페론 감마(IFN-γ)와 같은 사이토카인은 비드 서스펜션 분석 키트(bead suspension array kit, Bio-plex 200 system, Bio-Rad)를 이용하여 측정하였다. Cytokines such as interleukin (IL)-1β, IL-6, IL-10, IL-12, TNF-α (tumor necrosis alpha), and interferon gamma (IFN-γ) can be analyzed using a bead suspension assay kit. array kit, Bio-plex 200 system, Bio-Rad).
⑥ 간기능 검사 ⑥ Liver function test
⑥-1. 알라닌 아미노트랜스퍼라제 분석(Alanine aminotransferase (ALT) assay)⑥-1. Alanine aminotransferase (ALT) assay
간에서 총 ALT 농도는 ALT 색도계 분석 키트(ALT colorimetric assay kit, Biovision)에 의해 측정되었고, units(U/L)로 표현되었다. 원리에 의하면, 아미노 그룹은 알라닌에서 알파 케토글루타레이트(alpha ketoglutarate)로 전환된다. 가역적인 전환 반응을 통해 피루베이트(pyruvate)와 글루타메이트(glutamate)가 생성된다. 글루타메이트(Glutamate)는 535/587nm에서 흡광도가 측정되었다. Total ALT concentration in the liver was measured by ALT colorimetric assay kit (Biovision) and expressed in units (U/L). According to the principle, the amino group is converted from alanine to alpha ketoglutarate. Through a reversible conversion reaction, pyruvate and glutamate are produced. The absorbance of glutamate was measured at 535/587 nm.
⑥-2. 아스파테이트 아미노트랜스퍼라제 분석(Aspartate aminotransferase (AST or SGOT) assay)⑥-2. Aspartate aminotransferase (AST or SGOT) assay
간에서 총 AST 농도(concentration)는 AST 색도계 분석 키트(AST colorimetric assay kit, Biovision)에 의해 측정되었고 units(U/L)로 표현되었다. 원리에 의하면, 아미노 그룹(amino group)은 아스파테이트(aspartate)에서 알파 케토글루타레이트(alpha ketoglutarate)로 전환된다. 가역적인 전환 반응을 통해 옥살로아세테이트(oxaloacetate)와 글루타메이트(gluatamate)가 생성된다. 생성된 글루타메이트(glutamate)는 450nm에서 흡광도로 측정되었다. Total AST concentration in liver was measured by AST colorimetric assay kit (Biovision) and expressed in units (U/L). According to the principle, the amino group is converted from aspartate to alpha ketoglutarate. Through a reversible conversion reaction, oxaloacetate and glutamate are produced. Produced glutamate was measured by absorbance at 450 nm.
<실험예 2> 차아염소산 음용수의 안정성 분석결과<Experimental Example 2> Results of stability analysis of hypochlorous acid drinking water
<2-1> 차아염소산 음용수의 성분 분석<2-1> Component Analysis of Hypochlorous Acid Drinking Water
<실시예>에 따라 제조된 차아염소산 음용수를 공인기관(Eurofins Dr. Specht Laboratorien GmbH)에 의뢰하여 살충제 또는 인체 유해성분 550여 종을 검사하였으며, 그 결과 안전의 기준을 벗어나는 유해한 성분은 검출되지 않았다.Hypochlorous acid drinking water prepared according to <Example> was commissioned to an authorized institution (Eurofins Dr. Specht Laboratorien GmbH) to inspect 550 pesticides or harmful ingredients to the human body, and as a result, no harmful ingredients outside the safety standards were detected. .
<2-2> 체중과 간의 무게 측정 결과<2-2> Results of body weight and liver weight measurement
실험예 <1-2>에 따라 12주 동안 체중과 간의 무게 변화를 측정한 결과를 도 1에 나타내었다. Figure 1 shows the results of measuring changes in body weight and liver weight for 12 weeks according to Experimental Example <1-2>.
측정결과, 체중은 각 실험군 마다 일관성 있는 증가추세를 보여주었고, 실험군들 사이에 유의한 차이는 나타나지 않았다(도 1a). 간의 무게 또한 수돗물 음용군에 비해 차아염소산 음용군이 낮은 경향을 나타내었으나 유의한 차이는 보이지 않았다(도 1b).As a result of the measurement, the body weight showed a consistent increasing trend in each experimental group, and no significant difference was found between the experimental groups (Fig. 1a). The weight of the liver also tended to be lower in the hypochlorous acid drinking group than in the tap water drinking group, but no significant difference was observed (Fig. 1b).
<2-3> 활성산소 및 산화질소 측정 결과<2-3> Measurement results of active oxygen and nitric oxide
실험예 <1-4>의 ① 및 ②에 따라 혈청과 간 균질액에서 총 활성산소 및 산화질소의 양을 측정하였으며, 그 결과를 도 2에 나타내었다. According to ① and ② of Experimental Example <1-4>, the amounts of total active oxygen and nitric oxide were measured in serum and liver homogenate, and the results are shown in FIG. 2 .
측정결과, 활성산소의 경우, 혈청에서는 모든 그룹 사이에 활성산소 농도의 유의한 차이는 나타나지 않았으나, 간에서는 MS-차아염소산수 음용군이 미산성전기분해수 군에 비해 전체적으로 높은 경향을 보였고 고농도 MS-차아염소산수 음용군(p < 0.05)이 미산성전해수군에 비해 유의하게 높은 것으로 나타났다(도 2a). As a result of the measurement, in the case of active oxygen, there was no significant difference in active oxygen concentration between all groups in serum, but in the liver, the MS-hypochlorous acid water drinking group showed a tendency to be higher overall than the non-acidic electrolyzed water group, and the high-concentration MS- Hypochlorous acid water drinking group ( p < 0.05) was found to be significantly higher than the non-acidic electrolytic water group (Fig. 2a).
산화질소의 경우, 혈청 및 간 조직 내에서 그룹 간 농도의 유의한 차이를 나타내지 않았다(도 2b).In the case of nitric oxide, there was no significant difference in concentration between groups in serum and liver tissue (FIG. 2b).
<2-4> 항산화효소 Glutathione Peroxidase (GPx) 활성 및 항산화효소 카탈라제(Catalase) 활성 측정 결과<2-4> Measurement results of antioxidant enzyme Glutathione Peroxidase (GPx) activity and antioxidant enzyme catalase activity
실험예 <1-4>의 ③ 및 ④에 따라 혈청과 간 균질액에서 항산화효소 GPx 및 항산화효소 카탈라제의 농도를 측정하였으며, 그 결과를 도 3에 나타내었다. Concentrations of antioxidant enzyme GPx and antioxidant enzyme catalase were measured in serum and liver homogenate according to ③ and ④ of Experimental Example <1-4>, and the results are shown in FIG. 3 .
측정결과, 항산화효소 GPx의 경우, 혈청 GPx 농도는 실험군 사이에 유의한 차이를 나타내지 않았으나 간에서의 GPx 농도는 고농도 MS-차아염소산수 군이 가장 높게 나타났고 미산성전해수(p < 0.05), 저농도 MS-차아염소산수(p < 0.01), 중농도 MS-차아염소산수(p < 0.001)와 비교하여 유의한 차이를 나타내었다(도 3a). As a result of the measurement, in the case of the antioxidant enzyme GPx, the concentration of GPx in serum did not show a significant difference between the experimental groups, but the concentration of GPx in the liver was highest in the high-concentration MS-hypochlorous acid water group, and slightly acidic electrolyzed water ( p < 0.05), low-concentration GPx concentration Significant differences were shown compared to MS-hypochlorous acid water ( p < 0.01) and medium-concentration MS-hypochlorous acid water ( p < 0.001) (FIG. 3a).
카탈라제 활성의 경우, 간조직에서 모든 대조군과 실험군들 사이에 차이를 나타내지 않았으나, 혈청의 경우 미산성전해수(p < 0.01), 저농도 MS-차아염소산수(p < 0.001), 중농도 MS-차아염소산수(p < 0.001), 고농도 MS-차아염소산수(p < 0.001) 실험군들이 수돗물을 음용한 대조군에 비해 유의하게 낮게 나타났다(도 3b). In the case of catalase activity, there was no difference between all control and experimental groups in liver tissue, but in the case of serum, slightly acidic electrolytic water ( p < 0.01), low-concentration MS-hypochlorous acid water ( p < 0.001), medium-concentration MS-hypochlorous acid Water ( p < 0.001), high-concentration MS-hypochlorous acid water ( p < 0.001) experimental groups were significantly lower than the control group drinking tap water (Fig. 3b).
<2-5> 항산화효소 Glutathione Peroxidase (GPx) 활성 및 항산화효소 카탈라제(Catalase) 활성 측정 결과<2-5> Measurement results of antioxidant enzyme Glutathione Peroxidase (GPx) activity and antioxidant enzyme catalase activity
실험예 <1-4>의 ⑤에 따라 혈청과 간 균질액에서 염증성 사이토카인에 포함되는 IL-1β, IL-12, IFN-γ, TNF-α와 항염증성 사이토카인에 해당되는 IL-6와 IL-10의 농도를 측정하였으며, 그 결과를 도 4에 나타내었다.IL-1β, IL-12, IFN-γ, TNF-α included in inflammatory cytokines and IL-6 corresponding to anti-inflammatory cytokines in serum and liver homogenate according to ⑤ of Experimental Example <1-4> The concentration of IL-10 was measured, and the results are shown in FIG. 4 .
측정결과, MS-차아염소산수를 음용한 세 개의 실험군은 정수물과 수돗물 및 미산성전해수를 음용한 대조군과 비교하여 유의한 사이토카인 차이를 나타내지 않았다(도 4a 내지 도 4f).As a result of the measurement, the three experimental groups drinking MS-hypochlorous acid water did not show significant cytokine differences compared to the control group drinking purified water, tap water and non-acidic electrolytic water (FIGS. 4a to 4f).
<2-6> 간기능 지표 효소(ALT, AST) 측정 결과<2-6> Measurement results of liver function index enzymes (ALT, AST)
실험예 <1-4>의 ⑥에 따라 간에서 총 ALT 농도 및 총 AST의 농도를 측정하였으며, 그 결과를 도 5에 나타내었다.Total ALT concentration and total AST concentration were measured in the liver according to ⑥ of Experimental Example <1-4>, and the results are shown in FIG. 5 .
측정결과, 두 지표 모두 그룹간 유의한 차이를 나타내지 않았다(도 5).As a result of the measurement, both indicators did not show significant differences between groups (FIG. 5).
또한, 간의 조직학적 변화를 관찰하여 도 6에 나타내었다. 그 결과, 모든 실험군의 간조직은 핵의 모양과 크기, 염증성세포의 침윤, 미세구조 등을 포함하여 형태학적 차이를 나타내지 않았으며(도 6), 간독성과 관련된 병리학적 이상 소견을 보이지 않았다.In addition, the histological changes of the liver were observed and shown in FIG. 6 . As a result, the liver tissues of all experimental groups did not show morphological differences, including the shape and size of nuclei, infiltration of inflammatory cells, microstructure, etc. (FIG. 6), and did not show any pathological abnormalities related to liver toxicity.
<실험예 3> 차아염소산 음용수의 인체 적용 효과<Experimental Example 3> Effects of applying hypochlorous acid drinking water to the human body
2017년부터 유효염소 농도 50 ppm의 희석식 MS-차아염소산수(중농도 MS-차아염소산수, MS-HOCLM)를 제조하여 하루에 약 1.5 ~ 2L 분량을 음용하였으며, 음용 전과 음용 2년 후, 음용 3년 후 총 3회 건강검진을 통해 효과를 확인하였다.Since 2017, diluted MS-hypochlorous acid water (medium-concentration MS-hypochlorous acid water, MS-HOCL M ) with an effective chlorine concentration of 50 ppm has been manufactured and about 1.5 to 2 L of water has been consumed per day. Before drinking and 2 years after drinking, After 3 years of drinking, the effect was confirmed through a total of 3 health examinations.
검진결과는 하기 표 2 및 표 3에 나타난 바와 같다.The examination results are as shown in Tables 2 and 3 below.
기준범위standard range
기준범위standard range
여 50이상male over 40
female over 50
(mL/min/1.73m2)glomerular filtration rate
(mL/min/1.73m 2 )
여 38이하male under 56
female 38 or less
기준범위standard range
(전립선염)20-29
(prostatitis)
기준범위standard range
분석결과, i) 3년 동안 신장, 체중, 허리둘레, 체질량지수의 변화가 거의 없이 비슷한 수준을 유지하였다.As a result of the analysis, i) there was little change in height, weight, waist circumference, and body mass index for 3 years and maintained a similar level.
ii) 혈압의 경우, 음용 전 혈압은 138/80 mmHg로서 고혈압 상태였으나, 음용 2년 후 혈압은 128/78 mmHg, 음용 3년 후 120/68 mmHg로서 정상 혈압수준을 나타내었다.ii) In the case of blood pressure, the blood pressure before drinking was 138/80 mmHg, which was hypertensive, but after 2 years of drinking, the blood pressure was 128/78 mmHg and 120/68 mmHg after 3 years, indicating normal blood pressure levels.
iii) 혈액학적 검사 결과, 적혈구, 백혈구, 혈소판을 포함하여 음용 전과 음용 후의 검진 결과가 모두 정상 기준 범위 안에 포함되어 비슷한 수준을 유지하였다.iii) Hematological examination results, including red blood cells, white blood cells, and platelets, before and after drinking were all included within the normal standard range and maintained at similar levels.
iv) 소변을 이용한 요화학 검사 결과, 음용 전과 음용 후의 수치가 모두 정상범위 안에 포함되어 비슷한 수준을 유지하였다. 다만, 음용 대상자는 2018년 전립선염 감염으로 일시적으로 요침사가 증가하였다.iv) As a result of urine chemistry test using urine, the values before and after drinking were both within the normal range and maintained at similar levels. However, urinary sedimentation temporarily increased in drinking subjects due to prostatitis infection in 2018.
v) 혈청을 이용한 생화학적 검사 결과, 음용 전에는 총 콜레스테롤, 저밀도콜레스테롤, AST, ALT, 감마지티피가 정상범위를 훨씬 벗어났으나 MS-차아염소산수 음용 2년 후에는 정상 수준으로 개선되었다. 또한, 음용 3년 후에는 음용 2년 후와 비교하여 비슷하거나 더 향상된 결과를 나타내었다. 이 외의 다른 검사 항목의 경우에는 음용 전 후 3년 동안 비슷한 수준을 나타내었다.v) As a result of biochemical test using serum, total cholesterol, low-density cholesterol, AST, ALT, and gamma-GTP were far out of the normal range before drinking, but improved to normal levels after 2 years of drinking MS-hypochlorous acid water. In addition, after 3 years of drinking, similar or better results were shown compared to those after 2 years of drinking. Other test items showed similar levels before and after drinking for 3 years.
즉, MS-차아염소산수의 장기간 규칙적인 음용이 건강에 유해한 영향을 끼치지 않으며, 나아가 건강 증진에 도움이 되었음을 나타낸다.In other words, it indicates that regular drinking of MS-hypochlorous acid water for a long period of time did not adversely affect health, and furthermore, was helpful in improving health.
결론적으로, 본 발명에 따른 차아염소산 음용 조성물은 종래 차아염소산수와 달리 음용하더라도 안전성이 있다는 점이 충분이 검증되었으며, 건강 증진에 도움이 된다는 우수한 효과를 가진다.In conclusion, it has been sufficiently verified that the hypochlorous acid drinking composition according to the present invention is safe even when drunk unlike conventional hypochlorous acid water, and has an excellent effect of helping to improve health.
Claims (14)
상기 차아염소산염 혼합용액은 차아염소산염 용액 전체 중량대비 물 80 내지 99 중량부 및 차아염소산염 혼합물 1 내지 20 중량부이고,
상기 차아염소산염 혼합물은 차아염소산염 혼합물 전체 중량대비 차아염소산나트륨 70 내지 99 중량부 및 규소염 1 내지 30 중량부이며,
유효염소농도가 40 내지 60 ppm인, 음용 조성물.
It is prepared by mixing an organic acid mixture solution and a hypochlorite mixture solution with water,
The hypochlorite mixed solution is 80 to 99 parts by weight of water and 1 to 20 parts by weight of the hypochlorite mixture based on the total weight of the hypochlorite solution,
The hypochlorite mixture is 70 to 99 parts by weight of sodium hypochlorite and 1 to 30 parts by weight of silicon salt based on the total weight of the hypochlorite mixture,
A drinking composition having an effective chlorine concentration of 40 to 60 ppm.
상기 유기산 혼합용액은 아세트산, 구연산 및 개미산으로 구성된 그룹으로부터 선택된 1종 이상을 포함하는 것인, 음용 조성물.
According to claim 1,
The organic acid mixture solution is a drinking composition comprising at least one selected from the group consisting of acetic acid, citric acid and formic acid.
상기 유기산 혼합용액은 유기산 혼합용액 전체 중량대비 물 50 내지 99 중량부 및 유기산 혼합물 1 내지 50 중량부이고,
상기 유기산 혼합물은 유기산 혼합물 전체 중량대비 아세트산 40 내지 89 중량부, 구연산 10 내지 60 중량부 및 개미산 1 내지 20 중량부인, 음용 조성물.
According to claim 1,
The organic acid mixture solution is 50 to 99 parts by weight of water and 1 to 50 parts by weight of the organic acid mixture based on the total weight of the organic acid mixture solution,
The organic acid mixture is a drinking composition of 40 to 89 parts by weight of acetic acid, 10 to 60 parts by weight of citric acid and 1 to 20 parts by weight of formic acid based on the total weight of the organic acid mixture.
상기 음용 조성물은 음용 조성물 전체중량대비 유기산 혼합용액 0.01 내지 10 중량부, 차아염소산염 혼합용액 0.01 내지 10 중량부 및 물 80 내지 99.98 중량부인 음용 조성물.
According to claim 1,
The drinking composition comprises 0.01 to 10 parts by weight of an organic acid mixture solution, 0.01 to 10 parts by weight of a hypochlorite mixture solution, and 80 to 99.98 parts by weight of water, based on the total weight of the drinking composition.
상기 음용 조성물은 염소 부산물을 포함하지 않는 것인, 음용 조성물.
According to claim 1,
The drinking composition is a drinking composition that does not contain chlorine by-products.
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