KR101080842B1 - A method for extracting functional polysaccharides from seaweeds with high yield and the functional polysaccharides extracted by irradiation - Google Patents
A method for extracting functional polysaccharides from seaweeds with high yield and the functional polysaccharides extracted by irradiation Download PDFInfo
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- KR101080842B1 KR101080842B1 KR1020090001965A KR20090001965A KR101080842B1 KR 101080842 B1 KR101080842 B1 KR 101080842B1 KR 1020090001965 A KR1020090001965 A KR 1020090001965A KR 20090001965 A KR20090001965 A KR 20090001965A KR 101080842 B1 KR101080842 B1 KR 101080842B1
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- South Korea
- Prior art keywords
- seaweed
- polysaccharides
- seaweeds
- extracted
- irradiation
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Abstract
본 발명은 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법, 동 방법에 의해 추출된 다당류 및 동 방법에 의해 추출된 다당류의 사용방법에 관한 것으로 보다 상세하게는 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하는 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법, 동 방법에 의해 추출된 다당류 및 동 방법에 의해 추출된 다당류를 식품학적 조성물, 의약품 조성물 또는 화장품 조성물로 사용하는 방법에 관한 것이다.The present invention relates to a method for extracting polysaccharides from seaweeds using radiation, to a method for using polysaccharides extracted by the same method and polysaccharides extracted by the method, and more specifically, to extracting polysaccharides from seaweeds, A method of extracting polysaccharides from seaweeds using radiation irradiation during the process of extracting polysaccharides, using the polysaccharides extracted by the same method and the polysaccharides extracted by the same method as food composition, pharmaceutical composition or cosmetic composition It is about how to.
방사선 조사, 해조류, 다당류, 추출 Irradiation, Seaweed, Polysaccharides, Extraction
Description
본 발명은 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법, 동 방법에 의해 추출된 다당류 및 동 방법에 의해 추출된 다당류의 사용방법에 관한 것으로 보다 상세하게는 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하는 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법, 동 방법에 의해 추출된 다당류 및 동 방법에 의해 추출된 다당류를 식품학적 조성물, 의약품 조성물 또는 화장품 조성물로 사용하는 방법에 관한 것이다.The present invention relates to a method for extracting polysaccharides from seaweeds using radiation, to a method for using polysaccharides extracted by the same method and polysaccharides extracted by the method, and more specifically, to extracting polysaccharides from seaweeds, A method of extracting polysaccharides from seaweeds using radiation irradiation during the process of extracting polysaccharides, using the polysaccharides extracted by the same method and the polysaccharides extracted by the same method as food composition, pharmaceutical composition or cosmetic composition It is about how to.
해조류(sea algae)는 바다에서 나는 조류를 통틀어 이르는 말로서 조류가 자라는 바다의 깊이와 색깔에 따라 녹조류(green algae), 갈조류(brown algae), 홍조 류(red algae)로 나뉜다.Sea algae is a word that refers to algae from the sea and is divided into green algae, brown algae, and red algae according to the depth and color of the sea where the algae grows.
해조류는 각종 영양성분과 알긴산(alginic acid), 후코이단(Fucoidan), 카라기난(Carrageenan), 라미나린(laminarin) 등의 다당류와 클로로필(Chlorophyll) 등의 수용성 식이섬유의 장내기능, 칼슘과 칼륨 등의 무기질과 비타민과 같은 체내 신진 대사를 촉진시키는 물질들을 다량 함유하고 있기 때문에 현대인의 영양불균형이나 생리활성 성분을 보충할 수 있는 기능성 식품으로 기대된다. 이중에 특히 갈조류에 들어있는 주요성분인 후코이단은 유황을 함유하고 있어 항혈액응고 효과, 항암작용을 하며, 알긴산은 콜레스테롤을 감소시켜 동맥경화 예방, 고혈압 예방, 중금속 중독 예방을 하며, 라미나린은 혈압강하 작용으로 고혈압을 예방한다. 그리고 클로로필은 입냄새 제거로 청량감을 부여하고 중성 지방의 흡수를 저해, 대장암 예방, 변비 예방, 당뇨병 예방에 좋은 것으로 알려져 있다.Seaweed has various nutrients, intestinal function of polysaccharides such as alginic acid, fucoidan, carrageenan, laminarin, and water-soluble dietary fiber such as chlorophyll, minerals such as calcium and potassium. It is expected to be a functional food that can supplement the nutritional imbalance and physiologically active ingredients of modern people because it contains a large amount of substances that promote metabolism, such as vitamins and vitamins. Fucoidan, the main ingredient in brown algae, contains sulfur, which has anticoagulant effect and anticancer effect.Alginate reduces cholesterol and prevents arteriosclerosis, hypertension and heavy metal poisoning. Hypotension prevents hypertension. In addition, chlorophyll is known to be effective in removing odor and impairing the absorption of triglycerides, preventing colorectal cancer, preventing constipation, and preventing diabetes.
이러한 기능성 다당류를 해조류로부터 분리하기 위하여 여러 가지 추출 방법들이 개발되었다(Rioux LE, Turgeon SK, Beaulieu M (2007) Carbohydrate polymer 67:530-537; Hong Y, Wang K, Zhou C, Liu J, Zeng X (2008) Food Chemistry 111:428?432). 하지만, 해조류는 두터운 세포벽으로 둘러싸여있기 때문에 기능성 다당류의 추출 수율은 매우 낮다.Several extraction methods have been developed to separate these functional polysaccharides from seaweeds (Rioux LE, Turgeon SK, Beaulieu M (2007) Carbohydrate polymer 67: 530-537; Hong Y, Wang K, Zhou C, Liu J, Zeng X (2008) Food Chemistry 111: 428-432. However, because the algae are surrounded by a thick cell wall, the extraction yield of functional polysaccharides is very low.
이에 본 발명자는 해조류로부터 다당류의 추출방법에 대해 연구하던 중, 해조류에 감마선 및/또는 전자선과 같은 방사선을 조사하면 해조류의 세포벽이 파괴되어 알긴산, 후코이단, 카라기난, 라미나린 등의 다당류의 추출 수율이 증가한다는 것을 확인하고 본 발명을 완성하였다. 또한 해조류에 방사선을 조사하여 추출 수율이 증대한 다당류는 항산화, 항고혈압능 및 암세포에 대한 세포독성과 같은 기능성이 증가한다는 것을 확인하고 본 발명을 완성하였다.Therefore, while the present inventors are studying a method for extracting polysaccharides from seaweeds, irradiation of seaweeds with radiation such as gamma rays and / or electron beams destroys the cell walls of seaweeds, resulting in the extraction yield of polysaccharides such as alginic acid, fucoidan, carrageenan, and laminarin. It was confirmed that the increase was completed the present invention. In addition, polysaccharides of which the extraction yield was increased by irradiation with seaweeds were confirmed to increase functionalities such as antioxidant, antihypertensive activity and cytotoxicity against cancer cells, and completed the present invention.
본 발명의 목적은 방사선 조사를 이용하여 해조류로부터 고수율의 다당류를 추출하는 방법을 제공하고자 한다.An object of the present invention is to provide a method for extracting a high yield of polysaccharides from seaweed using irradiation.
본 발명의 다른 목적은 방사선 조사를 이용하여 해조류로부터 고수율의 다당류를 추출하는 방법에 의해 고수율로 추출된 다당류를 제공하고자 한다.Another object of the present invention is to provide a polysaccharide extracted in high yield by a method of extracting a high yield of polysaccharides from seaweed using radiation.
본 발명의 또 다른 목적은 방사선 조사를 이용하여 해조류로부터 고수율의 다당류를 추출하고, 이러한 다당류는 항산화, 항고혈압능 및 암세포에 대한 세포독성과 같은 기능성이 증대되어 식품학적 조성물, 의약품 조성물 또는 화장품 조성물로 사용하는 방법을 제공하고자 한다.Another object of the present invention is to extract a high yield of polysaccharides from seaweed using radiation, and these polysaccharides have increased functionality such as antioxidant, antihypertensive activity and cytotoxicity against cancer cells, so that the food composition, pharmaceutical composition or cosmetics It is intended to provide a method of use in the composition.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 고수율의 다당류를 추출하는 방법을 제공하고자 한다.The present invention, in the extraction of polysaccharides from seaweeds, to provide a method for extracting a high yield of polysaccharides from seaweeds by irradiation with radiation during the process of extracting polysaccharides from seaweeds.
본 발명에 따른 방사선 조사 기술을 이용하여 해조류로부터 높은 수율로 다당류를 추출하는 방법과 본 발명의 방사선 조사 기술을 이용하여 고수율로 추출된 다당류는 항산화활성, 항고혈압 및 암세포에 대한 세포독성과 같은 기능성이 증가 하는 등의 우수한 생물학적 특성을 보여 식품, 의약품 및 화장품 소재로 유용하게 사용될 수 있다.The method of extracting polysaccharides from seaweeds in high yield using the radiation technology according to the present invention and the polysaccharides extracted in high yield using the radiation technology of the present invention are characterized by antioxidant activity, antihypertension and cytotoxicity against cancer cells. Excellent biological properties such as increased functionality can be usefully used as food, pharmaceutical and cosmetic materials.
본 발명은 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법을 나타낸다.The present invention shows a method of extracting polysaccharides from seaweed using irradiation.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법을 나타낸다.In the present invention, in the extraction of polysaccharides from seaweeds, a method of extracting polysaccharides from seaweeds by irradiating with radiation during the step of extracting polysaccharides from seaweeds.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 다당류를 고수율로 추출하는 방법을 나타낸다.In the present invention, in the extraction of polysaccharides from seaweeds, a method of extracting polysaccharides from seaweeds in high yield by performing irradiation during the step of extracting polysaccharides from seaweeds.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 항산화활성, 항고혈압 및 암세포에 대한 세포독성과 같은 기능성이 증가된 다당류를 고수율로 추출하는 방법을 나타낸다.In the present invention, in the extraction of polysaccharides from seaweed, radiation is extracted during the process of extracting polysaccharides from seaweed to extract polysaccharides with increased functionalities such as antioxidant activity, antihypertension and cytotoxicity against cancer cells from seaweeds in high yield. The method is shown.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 항산화활성, 항고혈압 및 암세포에 대한 세포독성과 같은 기능성이 증가된 알긴산(alginic acid), 후코이단(Fucoidan), 카라기난(Carrageenan), 라미나린(laminarin)의 군으로부터 선택된 어느 하나 이상의 다당류를 고수율로 추출하는 방법을 나타낸다.In the present invention, in the extraction of polysaccharides from seaweeds, alginic acid, fucoidan, which has increased functionalities such as antioxidative activity, antihypertension, and cytotoxicity against cancer cells by irradiating with radiation during the extraction of polysaccharides from seaweeds (Fucoidan), carrageenan (Carrageenan), laminarin (laminarin) any one or more polysaccharides selected from the group of the high yield extraction method.
상기에서 해조류는 녹조류, 갈조류, 홍조류의 군으로부터 선택된 어느 하나 이상을 사용하여, 해조류로부터 다당류를 추출할 수 있다.The algae in the above can be used to extract polysaccharides from the algae using any one or more selected from the group of green algae, brown algae, red algae.
상기 해조류중 녹조류는 파래, 청각, 청태의 군으로부터 선택된 어느 하나 이상을 사용할 수 있다.The algae in the algae may be any one or more selected from the group of blue, auditory, jungtae.
상기 갈조류는 톳, 미역, 다시마, 대황, 모자반의 군으로부터 선택된 어느 하나 이상을 사용할 수 있다. The brown alga can be any one or more selected from the group of 톳, wakame seaweed, kelp, rhubarb, mabanban.
상기 홍조류는 우뭇가사리, 김, 카라니긴의 군으로부터 선택된 어느 하나 이상을 사용할 수 있다.The red alga may be any one or more selected from the group of wormwood, seaweed, and carrageenan.
상기에서 다당류는 알긴산, 후코이단, 카라기난, 포피란, 라미나린의 군으로부터 선택된 어느 하나 이상을 방사선 조사를 이용하여 해조류로부터 추출할 수 있다.The polysaccharide may be extracted from seaweeds using any one or more selected from the group of alginic acid, fucoidan, carrageenan, poryran, laminarin.
상기에서 해조류를 추출하는 공정 중에 조사하는 방사선은 감마선 또는 전자선을 사용할 수 있다.The radiation irradiated during the process of extracting seaweed may use gamma rays or electron beams.
상기에서 방사선 조사는 해조류를 추출하는 공정 중 해조류에 흡수되는 흡수선량이 1∼500kGy이 되도록 실시할 수 있다.In the above-mentioned irradiation, irradiation may be performed so that the absorbed dose absorbed by the seaweed is 1 to 500 kGy during the process of extracting the seaweed.
상기에서 방사선 조사는 해조류를 추출하는 공정 중 해조류에 흡수되는 흡수선량이 1∼300kGy이 되도록 실시할 수 있다.In the above-mentioned irradiation, irradiation may be performed so that the absorbed dose absorbed by the seaweeds is 1 to 300 kGy during the process of extracting the seaweeds.
상기에서 방사선 조사는 해조류를 추출하는 공정 중 해조류에 흡수되는 흡수선량이 1∼100kGy이 되도록 실시할 수 있다.In the above-mentioned irradiation, irradiation may be performed so that the absorbed dose absorbed by the seaweeds is 1 to 100 kGy during the process of extracting the seaweeds.
상기에서 방사선 조사는 해조류를 추출하는 공정 중 해조류에 흡수되는 흡수선량이 5∼50kGy이 되도록 실시할 수 있다.Irradiation may be carried out so that the absorbed dose absorbed by the algae during the process of extracting the algae is 5 to 50 kGy.
상기에서 해조류를 추출하는 공정 중 방사선 조사에 의해 해조류로부터 추출하는 다당류는 항산화의 기능성이 있다.The polysaccharide extracted from seaweeds by irradiation in the process of extracting seaweeds has the function of antioxidant.
상기에서 해조류를 추출하는 공정 중 방사선 조사에 의해 해조류로부터 추출하는 다당류는 항고혈압능의 기능성이 있다.The polysaccharide extracted from seaweeds by irradiation in the process of extracting seaweeds has the function of antihypertensive ability.
상기에서 해조류를 추출하는 공정 중 방사선 조사에 의해 해조류로부터 추출하는 다당류는 암세포에 대한 세포독성의 기능성이 있다.The polysaccharide extracted from the algae by irradiation in the process of extracting the algae has the functionality of cytotoxicity against cancer cells.
상기에서 해조류를 추출하는 공정 중 방사선 조사에 의해 해조류로부터 추출하는 다당류는 항산화, 항고혈압능 및 암세포에 대한 세포독성의 기능성이 있다.The polysaccharide extracted from seaweeds by irradiation in the process of extracting seaweeds has the function of antioxidant, antihypertensive capacity and cytotoxicity against cancer cells.
상기에서 해조류를 추출하는 공정 중 방사선을 조사하여 해조류로부터 다당류의 추출에 있어서 해조류를 추출하는 공정은 적어도 하나 이상의 유기용매를 이용한 유기용매 추출법에 의해 실시할 수 있다.The step of extracting the seaweed in the extraction of polysaccharides from the seaweed by irradiation with radiation during the step of extracting the seaweed can be carried out by an organic solvent extraction method using at least one organic solvent.
상기 유기용매 추출법에서의 유기용매는 탄소수가 1 내지 10개인 알코올 용매를 사용할 수 있다. 이러한 알코올 용매의 일예로 에탄올(ethanol)을 사용할 수 있다.The organic solvent in the organic solvent extraction method may be an alcohol solvent having 1 to 10 carbon atoms. Ethanol may be used as an example of such an alcohol solvent.
상기 유기용매 추출법에서의 유기용매는 염화물 용매를 사용할 수 있다. 이러한 염화물 용매의 일예로 칼시움 클로라이드(calcium chloride)를 사용할 수 있다. In the organic solvent extraction method, a chloride solvent may be used. An example of such a chloride solvent may be used calcium chloride.
상기에서 유기용매 추출법에서의 유기용매는 산(acid) 용매를 사용할 수 있 다. 이러한 산 용매의 일예로 염산(HCl), 황산, 아세트산의 군으로부터 선택된 어느 하나 이상의 산을 사용할 수 있다.The organic solvent in the organic solvent extraction method may be an acid solvent. As an example of such an acid solvent, any one or more acids selected from the group of hydrochloric acid (HCl), sulfuric acid, and acetic acid may be used.
상기 유기용매 추출법에서의 유기용매는 탄소수가 1 내지 10개인 알코올 용매, 염화물 용매, 산(acid) 용매의 군으로부터 선택된 어느 하나 이상을 사용할 수 있다.The organic solvent in the organic solvent extraction method may be used any one or more selected from the group of alcohol solvents, chloride solvents, acid solvents having 1 to 10 carbon atoms.
상기에서 방사선 조사를 이용하여 해조류로부터 다당류를 추출하는 일예로서 유기용매 추출법을 이용한 해조류로부터 다당류의 추출은 해조류에 해조류 중량 대비 5∼15배의 에탄올을 가한 다음 65∼75℃에서 3∼7시간 동안 환류추출을 1∼3회 실시한 후 여과한 다음 남은 잔사에 흡수선량이 1∼500kGy이 되도록 방사선 조사를 한다. 방사선 조사를 실시한 상기 잔사에 해조류 중량 대비 5∼15배의 칼슘 클로라이드(calcium chloride) 용액을 가한 후 65∼75℃에서 1∼5시간 동안 1∼5회 반복 추출하여 조 라미나린 층을 얻는다. 조 라미나린 추출 후 남은 잔사에 농도가 0.005∼0.02M이고 해조류 중량 대비 5∼15배인 염산 용액을 가하여 65∼75℃에서 1∼5시간 동안 1∼5회 반복 추출하여 조 후코이단 층을 얻는다. 상기 해조류로부터 추출한 조 라미나린 층, 조 후코이단 층은 각각 GPC(gel permeation chromatography)를 이용하여 분획하고 회수하여 해조류로부터 다당류인 라미나린, 후코이단 등을 추출할 수 있다. 이때 상기의 방사선 조사는 상기에서 언급한 방사선 조사 공정 대신에 해조류를 에탄올로 추출하기 전의 공정 또는 조 라미나린 층을 얻은 후 남은 잔사에 염산 용액을 가하기 전의 공정에서 실시할 수 있다. 또한 상기의 방사선 조사는 해조류를 에탄올에 추출하기 전, 해조류를 에탄올에 추출한 후, 조 라미나린 층을 얻은 후의 공정에서 동시에 실시할 수 있다.As an example of extracting polysaccharides from seaweeds using radiation irradiation, polysaccharides are extracted from seaweeds using an organic solvent extraction method by adding ethanol 5-15 times to seaweed weight to seaweeds for 3-7 hours at 65-75 ° C. After reflux extraction is carried out 1 ~ 3 times, it is filtered and irradiated with the remaining residue so that absorbed dose is 1 ~ 500kGy. To the residue subjected to irradiation was added 5-15 times the calcium chloride (calcium chloride) solution to the weight of the seaweed and then repeatedly extracted 1 to 5 times at 65 to 75 ℃ for 1 to 5 times to obtain a crude laminarin layer. After the crude laminarin extraction, the remaining residue was added with a hydrochloric acid solution having a concentration of 0.005 to 0.02 M and 5 to 15 times the weight of the seaweed, and extracted 1 to 5 times at 65 to 75 ° C. for 1 to 5 times to obtain a crude fucoidan layer. The crude laminarin layer and crude fucoidan layer extracted from the algae may be fractionated and recovered using GPC (gel permeation chromatography), respectively, to extract laminarin, fucoidan, and the like, from the algae. In this case, the irradiation may be performed in a step before the extraction of seaweeds with ethanol or in the step before adding the hydrochloric acid solution to the remaining residue after obtaining the crude laminarin layer instead of the above-mentioned irradiation step. In addition, the above irradiation can be carried out simultaneously in the step after the extraction of seaweeds in ethanol, after the extraction of seaweeds in ethanol, after obtaining a crude laminarin layer.
상기에서 해조류를 추출하는 공정 중 방사선 조사를 실시한 해조류로부터 다당류의 추출은 열수를 이용한 열수 추출법에 의해 실시할 수 있다.The extraction of polysaccharides from the seaweeds irradiated with radiation during the step of extracting seaweed can be carried out by the hot water extraction method using hot water.
상기에서 방사선 조사를 이용하여 해조류로부터 다당류를 추출하는 일예로서 열수 추출법을 이용한 해조류로부터 다당류의 추출은 해조류에 흡수선량이 1∼500kGy이 되도록 방사선 조사를 한 후 해조류 중량 대비 5∼15배의 정제수를 가한 다음 90∼120℃에서 1∼7시간 동안 열수 추출을 실시하여 열수 추출물을 얻은 후 이를 원심분리한 다음 에탄올, 메탄올과 같은 알코올이나 에틸아세트와 같은 유기 용매를 첨가하여이로부터 알긴산, 후코이단, 카라기난, 포피란, 라미나린의 군으로부터 선택된 어느 하나 이상의 다당류를 침전시켜 회수하여 얻을 수 있다.As an example of extracting polysaccharides from seaweeds using radiation irradiation, extraction of polysaccharides from seaweeds using hot water extraction is performed by irradiating seaweeds with an absorbed dose of 1 to 500 kGy and then using purified water 5 to 15 times the weight of seaweeds. After adding hot water extract at 90-120 ℃ for 1-7 hours to obtain hot water extract, centrifuged it, and then adding alcohol such as ethanol and methanol or organic solvent such as ethyl acetate to obtain alginic acid, fucoidan, carrageenan, Poppy is obtained by precipitating and recovering any one or more polysaccharides selected from the group of laminarins.
상기에서 방사선 조사를 이용하여 해조류로부터 다당류를 추출하는 일예로서 열수 추출법을 이용한 해조류로부터 다당류의 추출은 해조류에 흡수선량이 1∼500kGy이 되도록 방사선 조사를 한 후 해조류 중량 대비 5∼15배의 정제수를 가한 다음 최초 정제수의 부피 대비 10∼50％가 되도록 열수 추출을 실시하여 열수 추출물을 얻은 후 이를 원심분리한 다음 에탄올, 메탄올과 같은 알코올이나 에틸아세트와 같은 유기 용매를 첨가하여 이로부터 알긴산, 후코이단, 카라기난, 포피란, 라미나린의 군으로부터 선택된 어느 하나 이상의 다당류를 침전시켜 회수하여 얻을 수 있다.As an example of extracting polysaccharides from seaweeds using radiation irradiation, extraction of polysaccharides from seaweeds using hot water extraction is performed by irradiating seaweeds with an absorbed dose of 1 to 500 kGy and then using purified water 5 to 15 times the weight of seaweeds. After the addition of hot water extract to obtain 10 to 50% of the volume of the first purified water to obtain a hot water extract, centrifuged it, and then added an alcohol such as ethanol, methanol or an organic solvent such as ethyl acetate from the alginic acid, fucoidan, It can be obtained by precipitating and recovering any one or more polysaccharides selected from the group of carrageenan, porphyran and laminarin.
본 발명은 상기에서 언급한 방법인 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 포함한다.The present invention includes a polysaccharide obtained by a method of extracting polysaccharides from seaweeds by performing radiation in the process of extracting polysaccharides from seaweeds in the extraction of polysaccharides from seaweeds, which is the above-mentioned method.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 해조류에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 식품학적 조성물로 사용하는 방법을 나타낸다.In the present invention, in the extraction of polysaccharides from seaweeds, a method of using the polysaccharides obtained by the method of extracting polysaccharides from seaweeds by irradiating the seaweeds during the step of extracting the polysaccharides from the seaweeds as a food composition.
본 발명에 의한 상기 해조류에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 식품학적 조성물로 사용하는 예로서는 음료류, 면류, 냉동식품, 유가공 제품, 육가공 제품, 조미식품, 생식류 등과 같은 식품 조성물을 예로 들 수 있으나, 이에 한정되는 것은 아니다. Examples of using the polysaccharide obtained by the method of extracting the polysaccharide from the seaweed by irradiating the seaweed according to the present invention as a food composition include beverages, noodles, frozen food, dairy products, processed meat, seasoned food, raw food, etc. The same food composition may be exemplified, but is not limited thereto.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 해조류에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 의약품 조성물로 사용하는 방법을 나타낸다.In the present invention, in the extraction of polysaccharides from seaweeds, a method of using the polysaccharides obtained by the method of extracting polysaccharides from seaweeds by irradiating the seaweeds during the step of extracting the polysaccharides from the seaweeds as a pharmaceutical composition.
본 발명에 의한 상기 해조류에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 의약품 조성물로 사용하는 예로서는 정제, 과랍제, 환제, 액제, 주사제, 크림제, 연고제 등의 제형인 의약품 조성물이 바람직하나, 이에 한정되는 것은 아니다. Examples of using the polysaccharide obtained by the method of extracting the polysaccharide from the seaweed by irradiating the seaweed according to the present invention as a pharmaceutical composition include pharmaceutical products in the form of tablets, honeysuckle, pills, liquids, injections, creams, ointments, etc. Compositions are preferred, but are not limited to these.
본 발명은 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 해조류에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 화장품 조성물로 사용하는 방법을 나타낸다.This invention shows the method of using the polysaccharide obtained by the method of extracting polysaccharides from seaweeds by irradiating seaweeds in the process of extracting polysaccharides from seaweeds, and extracting polysaccharides from seaweeds as a cosmetic composition.
본 발명에 의한 해조류에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 화장품 조성물로 사용하는 예로서는 로션, 크림, 젤 등의 제형인 화장품 조성물이 바람직하나, 이에 한정되는 것은 아니다. As an example of using the polysaccharide obtained by the method of extracting polysaccharides from the seaweed by irradiating the seaweed according to the present invention as a cosmetic composition, a cosmetic composition, such as a lotion, cream, gel, or the like is preferable, but is not limited thereto.
상기에서 본 발명인 해조류로부터 다당류의 추출에 있어서, 해조류로부터 다당류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류는 항산화, 항고혈압능 및 암세포에 대한 세포독성 등의 기능성이 증가되는 특성을 나타내어, 이러한 다당류를 식품학적 조성물, 의약품 조성물 및/또는 화장품 조성물에 사용함으로써 상기의 기능성이 증가된 식품학적 조성물, 의약품 조성물 및/또는 화장품 조성물을 얻을 수 있다.In the extraction of polysaccharides from the seaweeds of the present invention, the polysaccharides obtained by the method of extracting polysaccharides from seaweeds by irradiation during the process of extracting the polysaccharides from the seaweeds have antioxidant, antihypertensive activity and cytotoxicity against cancer cells. By exhibiting the property of increasing functionality, such polysaccharides can be used in food, pharmaceutical and / or cosmetic compositions to obtain food, pharmaceutical and / or cosmetic compositions with increased functionality.
본 발명의 해조류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 식품학적 조성물, 화장품 조성물 및/또는 의약품 조성물로 사용하는 방법은 식품공전, 식품첨가물공전, 화장품 원료지정과 기준 및 시험방법 등에 관한 규정 등에서 허용되는 범위 내에서 필요에 따라 다양하게 변형하여 사용될 수 있다.The method of using the polysaccharide obtained by the method of extracting the polysaccharide from the seaweed by irradiation with radiation during the step of extracting the seaweed of the present invention as a food composition, a cosmetic composition and / or a pharmaceutical composition is a food formulation, food additives, cosmetics Various modifications may be made as necessary within the limits allowed by the regulations on the specification of raw materials, standards, test methods, etc.
상기에서 해조류를 추출하는 공정 중에 방사선 조사를 실시하여 해조류로부터 다당류를 추출하는 방법에 의해 얻은 다당류를 포함하는 식품학적 조성물, 화장품 조성물 또는 의약품 조성물의 제조방법은 특별한 제한없이, 통상의 방법으로 제조할 수 있다. The method of preparing a food composition, cosmetic composition or pharmaceutical composition comprising the polysaccharide obtained by the method of extracting the polysaccharide from the seaweed by irradiation with radiation during the step of extracting the seaweed can be prepared by conventional methods without particular limitation. Can be.
본 발명의 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법, 동 방법에 의해 추출된 다당류 및 동 방법에 의해 추출된 다당류의 사용방법에 대해 다양한 조건에 의해 조사한바, 본 발명의 목적을 달성하기 위해서는 상기에서 언급한 조건에 의해 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법, 동 방법에 의해 추출된 다당류 및 동 방법에 의해 추출된 다당류의 사용방법을 제공하는 것이 바람직하다.In order to achieve the object of the present invention, a method of extracting polysaccharides from seaweed using radiation of the present invention, polysaccharides extracted by the same method, and methods of using the polysaccharides extracted by the same method have been investigated under various conditions. It is desirable to provide a method of extracting polysaccharides from seaweeds using radiation under the above-mentioned conditions, a polysaccharide extracted by the same method, and a method of using the polysaccharide extracted by the same method.
이하 본 발명의 내용을 실시예 및 시험예를 통하여 구체적으로 설명한다. 그러나, 이들은 본 발명을 보다 상세하게 설명하기 위한 것으로 본 발명의 권리범위가 이들에 의해 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail with reference to Examples and Test Examples. However, these are intended to explain the present invention in more detail, and the scope of the present invention is not limited thereto.
<실험예 1> 해조류의 방사선 조사 방법Experimental Example 1 Irradiation Method of Seaweeds
본 발명에서 사용된 미역은 전라남도 완도 지역에서 회수하였다. 미역은 여분의 물기를 제거하여 다당류를 추출하거나 방사선 조사 후 다당류를 추출하였다.Seaweed used in the present invention was recovered in the Wando area of Jeollanam-do. Seaweed extracted polysaccharides by removing excess water or polysaccharides after irradiation.
또한 본 발명에서 사용된 다시마는 여분의 물기를 제거하여 다당류를 추출하거나 방사선 조사 후 다당류를 추출하였다.In addition, the kelp used in the present invention was extracted by extracting polysaccharides by removing excess water or polysaccharides after irradiation.
또한 본 발명에서 사용된 톳는 여분의 물기를 제거하여 다당류를 추출하거나 방사선 조사 후 다당류를 추출하였다.In addition, 톳 used in the present invention to remove the excess water to extract a polysaccharide or to extract a polysaccharide after irradiation.
또한 본 발명에서 사용된 모자반는 여분의 물기를 제거하여 다당류를 추출하거나 방사선 조사 후 다당류를 추출하였다.In addition, the mother board used in the present invention is to remove the excess water to extract a polysaccharide or to extract a polysaccharide after irradiation.
미역, 다시마, 톳 또는 모자반 시료는 한국원자력연구원 정읍방사선과학연구 소의 cobalt-60 irradiator에서 조사되었다. 선원의 크기는 약 11.1 BPq이었으며 선량율은 시간당 10kGy이었다. 흡수선량 확인은 5mm diameter alanine dosimeters(Bruker Instruments, Rheinstetten, Germany)로 하였으며, Dosimetry 시스템은 국제원자력기구(IAEA)의 규격에 준용하여 표준화한 후 사용하였으며, 총 흡수선량의 오차는 2％ 이내였다. 적용된 조사선량은 실온에서 0kGy, 5kGy, 10kGy, 30kGy, 50kGy로 조사되었다.Wakame, seaweed, seaweed, or mother-of-pearl samples were examined in a cobalt-60 irradiator at Korea Atomic Energy Research Institute, Jeongeup Radiation Science Institute. The source size was about 11.1 BPq and the dose rate was 10 kGy per hour. The absorbed dose was confirmed by 5mm diameter alanine dosimeters (Bruker Instruments, Rheinstetten, Germany). The Dosimetry system was used after standardizing in accordance with the International Atomic Energy Agency (IAEA) standard, and the total absorbed dose error was within 2%. The applied dose was irradiated at 0kGy, 5kGy, 10kGy, 30kGy, 50kGy at room temperature.
전자선 조사는 ELV4-electron accelerator(Energy 10 MeV, beam power 570 kW)를 이용하여 감마선 조사와 동일한 총 흡수선량을 갖도록 하였다. 이때 에너지는 10 MeV이고 전류(current)는 1mA 였다. 그리고 낮은 투과력 때문에 전자선 조사되는 해조류 시료의 두께는 3mm로 하여 조사하였다. 감마선 및 전자선 조사된 해조류는 4℃에 저장하면서 향 후 실험에 사용하였다.Electron beam irradiation was performed using ELV4-electron accelerator (Energy 10 MeV, beam power 570 kW) to have the same total absorbed dose as gamma radiation. At this time, the energy was 10 MeV and the current was 1 mA. In addition, the thickness of the seaweed samples irradiated with electron beams was 3 mm because of low permeability. Gamma and electron beam irradiated seaweed was used for future experiments while stored at 4 ℃.
<실험예 2> 해조류의 다당류 추출 방법Experimental Example 2 Polysaccharide Extraction Method of Seaweeds
미역 내 함유된 기능성 다당류인 알긴산, 카라긴산, 후코이단 및 라미나린은 Rioux (Rioux LE, Turgeon SL, Beaulieu M (2007) Carbohydrate Polymers 69(3): 530-537) 방법을 변형하여 이용하였다. The functional polysaccharides alginate, carrageic acid, fucoidan and laminarin contained in seaweed were used by modifying the Rioux (Rioux LE, Turgeon SL, Beaulieu M (2007) Carbohydrate Polymers 69 (3): 530-537) method.
시료인 미역 60g에 85％ 에탄올 600ml을 가한 다음 70℃에서 5시간 동안 환류추출(2회 반복)을 한 후, 여과지(Whatman filter paper No. 4)를 이용하여 여과하였다. 남은 잔사에 2％ calcium chloride 용액 600ml을 가한 후 70℃에서 3시간 동안 3회반복 추출하여 조 라미나린 층을 얻었다. 조 라미나린 추출 후 남은 잔사 에 0.01M 염산 용액 600ml을 가하여 70℃에서 3시간 동안 3회반복 추출하여 조 후코이단 층을 얻었다. 추출한 시료는 GPC(gel permeation chromatography)를 이용하여 분획하여 회수한 다음 48시간 동안 투석(cut off 1,000 Da)하였다. 투석 시료는 농축하여 동결건조한 후 냉장보관하여 실험에 사용하였다.600 ml of 85% ethanol was added to 60 g of seaweed, which was then refluxed (repeated twice) at 70 ° C. for 5 hours, and then filtered using a filter paper (Whatman filter paper No. 4). 600 ml of 2% calcium chloride solution was added to the remaining residue, and the mixture was extracted three times at 70 ° C. for 3 hours to obtain a crude laminarin layer. 600 ml of 0.01 M hydrochloric acid solution was added to the remaining residue after crude laminarin extraction, and the mixture was extracted three times at 70 ° C. for 3 hours to obtain a crude fucoidan layer. The extracted samples were collected by fractionation using gel permeation chromatography (GPC) and dialyzed (cut off 1,000 Da) for 48 hours. Dialysis samples were concentrated, lyophilized and refrigerated to be used in the experiment.
또한 후코이단 및 라미나린의 추출을 위하여 열수 추출 방법을 사용하였다. 95 ℃ 항온수조에서 60분간 교반 추출을 수행한 후 추출된 반응액을 여과나 원심분리하여 얻어진 추출액에 에탄올, 메탄올과 같은 알코올이나 에틸아세트와 같은 유기 용매를 첨가하여 후코이단 및 라미나린을 침전시켜 회수하고 수세 후 건조하였다.Also hot water extraction method was used for the extraction of fucoidan and laminarin. After 60 minutes of stirring and extraction in a constant temperature bath at 95 ℃, the extracted reaction solution was filtered or centrifuged to extract and recovered by extracting fucoidan and laminarin by adding an alcohol such as ethanol and methanol or an organic solvent such as ethyl acetate. And dried after washing with water.
<실시예 1-1>≪ Example 1-1 >
수세, 세척 후 표면의 물기를 제거한 해조류인 미역에 방사선으로 흡수선량이 10kGy이 되도록 감마선 조사를 실시하였다. Gamma-irradiation was performed so that the absorbed dose was 10 kGy by radiation to seaweed, which was seaweed removed from the surface after washing with water and washing.
상기 방사선 조사를 실시한 미역 시료 60g에 85％ 에탄올 600ml을 가한 다음 70℃에서 5시간 동안 환류추출을 2회 반복한 후, 여과지(Whatman filter paper No. 4)를 이용하여 여과하였다. 여과 후 남은 잔사에 2％ 칼슘 클로라이드(calcium chloride) 용액 600ml을 가한 후 70℃에서 3시간 동안 3회 반복 추출하여 조 라미나린 층을 얻었다. 600 ml of 85% ethanol was added to 60 g of the seaweed sample irradiated with radiation, followed by repeated reflux extraction at 70 ° C. for 5 hours, followed by filtering using a filter paper (Whatman filter paper No. 4). 600 ml of a 2% calcium chloride solution was added to the remaining residue after filtration, and the mixture was extracted three times at 70 ° C. for 3 hours to obtain a crude laminarin layer.
조 라미나린 추출 후 남은 잔사에 0.01M 염산 용액 600ml을 가하여 70℃에서 3시간 동안 3회 반복 추출하여 조 후코이단 층을 얻었다. 600 ml of 0.01 M hydrochloric acid solution was added to the remaining residue after the crude laminarin extraction, and extracted three times at 70 ° C. for 3 hours to obtain a crude fucoidan layer.
상기의 공정에서 얻은 조 라미나린 층 및 조 후코이단 층은 GPC(gel permeation chromatography)를 이용하여 분획하여 라미나린 및 후코이단을 회수한 다음 48시간 동안 투석(cut off 1,000 Da)하였다. 투석 시료는 완전 건조되도록 동결건조한 후 4℃ 온도의 냉장보관하였다.The crude laminarin layer and the crude fucoidan layer obtained in the above process were fractionated by gel permeation chromatography (GPC) to recover laminarin and fucoidan, and then dialyzed (cut off 1,000 Da) for 48 hours. The dialysis sample was lyophilized to dry completely and then refrigerated at 4 ° C.
<실시예 1-2>≪ Example 1-2 >
해조류로서 미역 대신 톳을 사용하는 것을 제외하고는 상기 실시예 1-1과 동일한 방법으로 톳으로부터 후코이단, 라미나린의 다당류를 추출하였다.Except for using seaweed instead of seaweed as seaweed, polysaccharides of fucoidan and laminarin were extracted from the seaweed in the same manner as in Example 1-1.
<실시예 1-3>≪ Example 1-3 >
해조류로서 미역 대신 모자반을 사용하는 것을 제외하고는 상기 실시예 1-1과 동일한 방법으로 모자반으로부터 후코이단, 라미나린의 다당류를 추출하였다.Polysaccharides of fucoidan and laminarin were extracted from the algae in the same manner as in Example 1-1, except that the algae was used instead of seaweed as seaweed.
<실시예 2-1><Example 2-1>
수세, 세척 후 표면의 물기를 제거한 해조류인 미역 시료 60g에 85％ 에탄올 600ml을 가한 다음 70℃에서 5시간 동안 환류추출을 2회 반복한 후, 여과지(Whatman filter paper No. 4)를 이용하여 여과하였다. 여과 후 남은 잔사에 방사선으로 흡수선량이 10kGy이 되도록 감마선 조사를 하고 이후 2％ 칼슘 클로라이드 용액 600ml을 가한 후 70℃에서 3시간 동안 3회 반복 추출하여 조 라미나린 층을 얻었다. After washing and washing, 600 ml of 85% ethanol was added to 60 g of seaweed seaweed sample, which was removed from the surface water, and then refluxed twice at 70 ° C. for 5 hours, followed by filtration using a filter paper (Whatman filter paper No. 4). It was. The remaining residue after filtration was irradiated with gamma radiation so that the absorbed dose was 10 kGy. Then, 600 ml of a 2% calcium chloride solution was added thereto, and the mixture was extracted three times at 70 ° C. for 3 hours to obtain a crude laminarin layer.
조 라미나린 추출 후 남은 잔사에 0.01M 염산 용액 600ml을 가하여 70℃에서 3시간 동안 3회 반복 추출하여 조 후코이단 층을 얻었다. 600 ml of 0.01 M hydrochloric acid solution was added to the remaining residue after the crude laminarin extraction, and extracted three times at 70 ° C. for 3 hours to obtain a crude fucoidan layer.
상기의 공정에서 얻은 조 라미나린 층 및 조 후코이단 층은 GPC(gel permeation chromatography)를 이용하여 분획하여 라미나린 및 후코이단을 회수한 다음 48시간 동안 투석(cut off 1,000 Da)하였다. 투석 시료는 완전 건조되도록 동결건조한 후 4℃ 온도의 냉장보관하였다.The crude laminarin layer and the crude fucoidan layer obtained in the above process were fractionated by gel permeation chromatography (GPC) to recover laminarin and fucoidan, and then dialyzed (cut off 1,000 Da) for 48 hours. The dialysis sample was lyophilized to dry completely and then refrigerated at 4 ° C.
<실시예 2-2><Example 2-2>
해조류로서 미역 대신 톳을 사용하는 것을 제외하고는 상기 실시예 2-1과 동일한 방법으로 톳으로부터 후코이단, 라미나린의 다당류를 추출하였다.Except for using seaweed instead of seaweed as seaweed, polysaccharides of fucoidan and laminarin were extracted from seaweed in the same manner as in Example 2-1.
<실시예 2-3><Example 2-3>
해조류로서 미역 대신 모자반을 사용하는 것을 제외하고는 상기 실시예 2-1과 동일한 방법으로 모자반으로부터 후코이단, 라미나린의 다당류를 추출하였다.The polysaccharides of fucoidan and laminarin were extracted from the alveolar in the same manner as in Example 2-1, except that the algae was used instead of seaweed as seaweed.
<실시예 3-1><Example 3-1>
수세, 세척 후 표면의 물기를 제거한 해조류인 미역 시료 60g에 85％ 에탄올 600ml을 가한 다음 70℃에서 5시간 동안 환류추출을 2회 반복을 한 후, 여과지(Whatman filter paper No. 4)를 이용하여 여과하였다. After washing and washing, 600 ml of 85% ethanol was added to 60 g of seaweed seaweed sample, which was removed from the surface of the seaweed, followed by repeated reflux extraction at 70 ° C. for 5 hours, using filter paper (Whatman filter paper No. 4). Filtered.
여과 후 남은 잔사에 2％ 칼슘 클로라이드 용액 600ml을 가한 후 70℃에서 3 시간 동안 3회 반복 추출하여 조 라미나린 층을 얻었다. 600 ml of a 2% calcium chloride solution was added to the remaining residue after filtration, and the mixture was extracted three times at 70 ° C. for 3 hours to obtain a crude laminarin layer.
조 라미나린 추출 후 남은 잔사에 방사선으로 흡수선량이 10kGy이 되도록 감마선 조사를 한 후에 0.01M 염산 용액 600ml을 가하여 70℃에서 3시간 동안 3회 반복 추출하여 조 후코이단 층을 얻었다. After irradiating gamma rays to the remaining residue after crude laminarin extraction by radiation to 10 kGy, 600 ml of 0.01 M hydrochloric acid solution was added and extracted three times at 70 ° C. for 3 hours to obtain crude fucoidan layer.
상기의 공정에서 얻은 조 라미나린 층 및 조 후코이단 층은 GPC(gel permeation chromatography)를 이용하여 분획하여 라미나린 및 후코이단을 회수한 다음 48시간 동안 투석(cut off 1,000 Da)하였다. 투석 시료는 완전 건조되도록 동결건조한 후 4℃ 온도의 냉장보관하였다.The crude laminarin layer and the crude fucoidan layer obtained in the above process were fractionated by gel permeation chromatography (GPC) to recover laminarin and fucoidan, and then dialyzed (cut off 1,000 Da) for 48 hours. The dialysis sample was lyophilized to dry completely and then refrigerated at 4 ° C.
<실시예 3-2><Example 3-2>
해조류로서 미역 대신 톳을 사용하는 것을 제외하고는 상기 실시예 3-1과 동일한 방법으로 톳으로부터 후코이단, 라미나린의 다당류를 추출하였다.Except for using seaweed instead of seaweed as seaweed, polysaccharides of fucoidan and laminarin were extracted from seaweed in the same manner as in Example 3-1.
<실시예 3-3><Example 3-3>
해조류로서 미역 대신 모자반을 사용하는 것을 제외하고는 상기 실시예 3-1과 동일한 방법으로 모자반으로부터 후코이단, 라미나린의 다당류를 추출하였다.Polysaccharides of fucoidan and laminarin were extracted from the algae in the same manner as in Example 3-1, except that the algae was used instead of seaweed as seaweed.
<실시예 4-1><Example 4-1>
방사선으로 흡수선량이 10kGy이 되도록 전자선 조사를 실험예 1을 참조하여 실시하는 것을 제외하고는 상기 실시예 1-1과 동일한 방법으로 해조류로부터 다당 류를 추출하였다.The polysaccharide was extracted from the seaweed in the same manner as in Example 1-1 except that the electron beam irradiation was carried out with reference to Experimental Example 1 so that the absorbed dose was 10 kGy.
<실시예 4-2><Example 4-2>
해조류로서 미역 대신 톳을 사용하는 것을 제외하고는 상기 실시예 4-1과 동일한 방법으로 톳으로부터 후코이단, 라미나린의 다당류를 추출하였다.Except for using seaweed instead of seaweed as seaweed, polysaccharides of fucoidan and laminarin were extracted from seaweed in the same manner as in Example 4-1.
<실시예 4-3><Example 4-3>
해조류로서 미역 대신 모자반을 사용하는 것을 제외하고는 상기 실시예 4-1과 동일한 방법으로 모자반으로부터 후코이단, 라미나린의 다당류를 추출하였다.Fucoidan and laminarin polysaccharides were extracted from the algae in the same manner as in Example 4-1, except that the algae was used instead of seaweed as seaweed.
<실시예 5-1><Example 5-1>
방사선으로 흡수선량이 10kGy이 되도록 전자선 조사를 실험예 1을 참조하여 실시하는 것을 제외하고는 상기 실시예 2-1과 동일한 방법으로 해조류로부터 다당류를 추출하였다.The polysaccharide was extracted from the algae in the same manner as in Example 2-1 except that the electron beam irradiation was carried out with reference to Experimental Example 1 so that the absorbed dose was 10 kGy.
<실시예 5-2>≪ Example 5-2 >
해조류로서 미역 대신 톳을 사용하는 것을 제외하고는 상기 실시예 5-1과 동일한 방법으로 톳으로부터 후코이단, 라미나린의 다당류를 추출하였다.Except for using seaweed instead of seaweed as seaweed, polysaccharides of fucoidan and laminarin were extracted from the seaweed in the same manner as in Example 5-1.
<실시예 5-3>≪ Example 5-3 >
해조류로서 미역 대신 모자반을 사용하는 것을 제외하고는 상기 실시예 5-1과 동일한 방법으로 모자반으로부터 후코이단, 라미나린의 다당류를 추출하였다.Polysaccharides of fucoidan and laminarin were extracted from the algae in the same manner as in Example 5-1, except that the algae was used instead of seaweed as seaweed.
<실시예 6-1><Example 6-1>
방사선으로 흡수선량이 10kGy이 되도록 전자선 조사를 실험예 1을 참조하여 실시하는 것을 제외하고는 상기 실시예 3-1과 동일한 방법으로 해조류로부터 다당류를 추출하였다.Polysaccharide was extracted from the seaweed in the same manner as in Example 3-1, except that the electron beam irradiation was carried out with reference to Experimental Example 1 so that the absorbed dose was 10 kGy.
<실시예 6-2><Example 6-2>
해조류로서 미역 대신 톳을 사용하는 것을 제외하고는 상기 실시예 6-1과 동일한 방법으로 톳으로부터 후코이단, 라미나린의 다당류를 추출하였다.Except for using seaweed instead of seaweed as seaweed, polysaccharides of fucoidan and laminarin were extracted from the seaweed in the same manner as in Example 6-1.
<실시예 6-3><Example 6-3>
해조류로서 미역 대신 모자반을 사용하는 것을 제외하고는 상기 실시예 6-1과 동일한 방법으로 모자반으로부터 후코이단, 라미나린의 다당류를 추출하였다.Polysaccharides of fucoidan and laminarin were extracted from the algae in the same manner as in Example 6-1, except that the algae was used instead of seaweed as seaweed.
<실시예 7> 방사선 조사를 이용하여 해조류로부터 추출된 다당류를 포함하는 식품학적 조성물Example 7 A Food Composition comprising Polysaccharides Extracted from Seaweed Using Irradiation
밀가루를 주재료로 하는 면 제조에 있어서, 상기 실시예 2-1에서 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단을 면 중량 대비 2％ 첨가하 는 단계를 포함하도록 하여 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단이 함유된 면을 제조하였다.In the manufacturing of cotton as a main ingredient, the method comprises adding 2% of fucoidan, which is a polysaccharide extracted from seaweed using radiation irradiation, to 2% by weight of cotton, from seaweed using radiation. Noodles containing fucoidan, an extracted polysaccharide, were prepared.
<실시예 8> 방사선 조사를 이용하여 해조류로부터 추출된 다당류를 포함하는 의약품 조성물Example 8 A pharmaceutical composition comprising a polysaccharide extracted from seaweed using irradiation
찰과상 치료용 외용제 연고 제조에 있어서, 상기 실시예 2-1에서 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단을 면 중량 대비 1％ 첨가하는 단계를 포함하도록 하여 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단이 함유된 찰과상 치료용 외용제 연고를 제조하였다.In preparing the external preparation ointment for abrasion treatment, adding 1% of fucoidan, which is a polysaccharide extracted from seaweed using radiation irradiation, to 1% by weight of cotton, was extracted from seaweed using radiation. A topical ointment for the treatment of abrasions containing the polysaccharide fucoidan was prepared.
<실시예 9> 방사선 조사를 이용하여 해조류로부터 추출된 다당류를 포함하는 화장품적 조성물Example 9 Cosmetic Composition Comprising Polysaccharides Extracted from Seaweed Using Irradiation
여성용 바디로션의 제조에 있어서, 상기 실시예 2-1에서 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단을 바디로션 총 중량 대비 2％가 되도록 첨가하는 단계를 이용하여 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단이 함유된 여성용 바디로션을 제조하였다.In the manufacture of women's body lotion, in Example 2-1, the polysaccharide extracted from seaweed using radiation is added to the fucoidan so that 2% of the total weight of the body lotion by using radiation. A female body lotion containing fucoidan, an extracted polysaccharide, was prepared.
<시험예 1><Test Example 1>
상기 실시예 1 내지 실시예 3으로부터 얻은 다당류와 대조구로서 방사선 조사를 하지 않고 실시예 1의 방법에 의해 얻은 다당류에 대한 추출 수율을 건조 해 조류의 질량에 대한 회수된 다당류의 질량 백분율로 측정하고 그 결과를 아래의 표 1에 나타내었다.The extraction yield of the polysaccharides obtained by the method of Example 1 without irradiation with the polysaccharides obtained from Examples 1 to 3 as a control was measured by the mass percentage of the recovered polysaccharides to the mass of algae The results are shown in Table 1 below.
표 1. 해조류로부터 방사선 조사 방법에 따른 기능성 다당류의 추출 수율Table 1. Extraction yield of functional polysaccharides from seaweed according to the irradiation method
*대조구는 방사선 흡수선량이 0kGy, 즉 방사선 조사를 하지 않는 것을 제외하고는 실시예 1과 동일한 방법에 의해 미역으로부터 후코이단 및 라미나린을 추출한 것이다.* The control tool extracts fucoidan and laminarin from seaweed by the same method as in Example 1 except that the radiation absorbed dose is 0 kGy, that is, no irradiation.
상기 표 1의 결과에서처럼 방사선 조사를 이용하여 해조류인 미역을 추출한 실시예 1 내지 실시예 3은 방사선 조사를 하지 않은 대조구에 비해 다당류인 후코이단, 라미나린의 추출 수율이 보다 우수함을 알 수 있었다.As shown in the results of Table 1, Examples 1 to 3 extracted seaweed seaweed using radiation irradiation was found to have a better extraction yield of polysaccharides fucoidan and laminarin than the control group not irradiated.
한편 표 1에서처럼 다당류의 하나인 후코이단 추출 수율은 후코이단 층을 얻기 전에 방사선 조사를 실시한 실시예 3이 제일 높았으며, 다당류의 다른 하나인 라미나린 추출 수율은 라미나린 층을 얻기 전에 방사선 조사를 실시한 실시예 2가 제일 높았음을 알 수 있었다. 이러한 결과는 해조류로부터 다당류를 추출함에 있어서, 원하는 종류의 다당류를 추출하기 전에 방사선 조사를 실시하는 것이 원하는 종류의 다당류 추출 수율에 보다 유리함을 알 수 있었다.Meanwhile, as shown in Table 1, the yield of fucoidan extraction, which is one of the polysaccharides, was highest in Example 3, which was irradiated before obtaining the fucoidan layer, and the yield of laminarin, which was the other polysaccharide, was irradiated before obtaining the laminarin layer. Example 2 was the highest. These results indicate that in the extraction of polysaccharides from seaweeds, it is more advantageous for the desired yield of polysaccharide extraction to perform irradiation before extracting the desired polysaccharides.
<시험예 2><Test Example 2>
상기 실시예 2에서 방사선 조사시 흡수선량이 0kGy, 5kGy, 10kGy, 30kGy, 50kGy이 되도록 하여 방사선 조사시 흡수선량에 따른 다당류의 추출 수율을 건조 해조류의 질량에 대한 회수된 다당류의 질량 백분율로 측정하고 그 결과를 아래의 표 2에 나타내었다.In Example 2, the absorbed dose was 0kGy, 5kGy, 10kGy, 30kGy, 50kGy when irradiated, and the extraction yield of polysaccharides according to the absorbed dose during irradiation was measured as the mass percentage of the recovered polysaccharides to the mass of the dried algae. The results are shown in Table 2 below.
표 2. 해조류로부터 방사선 조사 선량에 따른 기능성 다당류의 추출 수율Table 2. Extraction Yield of Functional Polysaccharides from Seaweeds with Irradiation Dose
상기 표 2에서 보이는 것처럼 방사선 조사 선량이 증가함에 따라 최종 얻어지는 후코이단이나 라미나린과 같은 기능성 다당류의 추출 수율이 증가함을 알 수 있었다.As shown in Table 2, it was found that the extraction yield of functional polysaccharides such as fucoidan and laminarin increased as the radiation dose increased.
한편, 도 1에 해조류인 미역에 방사선 조사를 한 세포의 Transmission Electron Microscope(TEM) 사진과 방사선 조사를 하지 않은 세포의 TEM 사진을 나타내었다. Meanwhile, FIG. 1 shows a Transmission Electron Microscope (TEM) photograph of a cell irradiated with seaweed, seaweed, and a TEM photograph of a non-irradiated cell.
도 1(a)는 방사선 조사를 하지 않은 해조류인 미역 세포의 TEM 사진이고, 도 1(b)는 실시예 2에서 10kGy 선량의 감마선을 조사한 해조류의 TEM 사진이고, 도 1(c)는 실시예 5에서 10kGy 선량의 전자선을 조사한 해조류의 TEM 사진이다. 한편 도 1에서 화살표는 세포벽을 나타낸다. Figure 1 (a) is a TEM picture of seaweed seaweed cells that are not irradiated seaweed, Figure 1 (b) is a TEM picture of algae irradiated with gamma rays of 10kGy dose in Example 2, Figure 1 (c) is an example TEM image of algae irradiated with 5 to 10 kGy electron beam. Meanwhile, the arrows in FIG. 1 indicate cell walls.
도 1(b), 도 1(c)는 방사선 조사에 의해 해조류인 미역 세포의 세포벽이 도 1(a)에 비해 얇아지거나 파괴된 것을 보이고 있다.1 (b) and 1 (c) show that the cell walls of seaweed seaweed cells become thinner or destroyed than those of FIG. 1 (a) by irradiation.
<시험예 3> 해조류 다당류의 항산화 측정Test Example 3 Antioxidant Measurement of Seaweed Polysaccharides
상기 실시예 2에서 방사선 조사시 흡수선량이 0kGy, 5kGy, 10kGy, 30kGy, 50kGy이 되도록 하여 방사선 조사시 흡수선량에 따라 추출한 후코이단과 라미나린의 DPPH 라디칼 제어능을 측정하여 방사선 조사를 한 해조류로부터 추출한 후코이단과 라미나린의 항산화 특성을 조사하고 그 결과를 아래의 표 3에 나타내었다. In Example 2, the absorbed dose was 0kGy, 5kGy, 10kGy, 30kGy, 50kGy when irradiated, and the DPPH radical control ability of fucoidan and laminarin extracted according to the absorbed dose during radiation was measured and extracted from the irradiated algae. The antioxidant properties of fucoidan and laminarin were investigated and the results are shown in Table 3 below.
상기에서 방사선 조사를 한 해조류로부터 추출된 다당류인 후코이단과 라미나린의 전자공여능 측정은 Blois의 방법(Amarowicz R, Pegg R B, Rahimi MP, Barl B, Weil JA (2004). Food Chemistry 84: 551-562.)에 준하여 다당류의 DPPH(2,2-diphenyl-1-picryl-hydrazil)에 대한 수소공여 효과로 측정하였다. 즉, 시료인 각각의 다당류 2ml에 2×10-4M DPPH용액(dissolved in 99％ Ethanol)을 1ml 가하고, 혼합(vortex mixing)하여 37℃에서 30분간 반응시켰다. 이 반응액을 517nm에서 분광광도계(UV 1600 PC, Shimadzu, Tokyo, Japan)를 이용하여 흡광도를 측정하였으며 전자공여 효과는 시료첨가 전,후의 흡광도 차이를 백분율(％)로 나타내었다.The electron donating ability of fucoidan and laminarine, polysaccharides extracted from irradiated seaweeds, was measured by Blois' method (Amarowicz R, Pegg RB, Rahimi MP, Barl B, Weil JA (2004) .Food Chemistry 84: 551-562 .) Was determined by the hydrogen donating effect on DPPH (2,2-diphenyl-1-picryl-hydrazil) of polysaccharides. That is, 1 ml of 2 × 10 −4 M DPPH solution (dissolved in 99% Ethanol) was added to 2 ml of each polysaccharide as a sample, and the mixture was reacted at 37 ° C. for 30 minutes by vortex mixing. The absorbance of the reaction solution was measured at 517 nm using a spectrophotometer (UV 1600 PC, Shimadzu, Tokyo, Japan). The electron donating effect was expressed as a percentage (%) of the absorbance difference before and after sample addition.
전자공여능(％)=[1 - (시료첨가구의 흡광도/시료무첨가구의 흡광도)]×100Electron donating ability (%) = [1-(absorbance of sample addition / absorbance of sample no addition)] × 100
상기 실시예 2에서 방사선 조사시 흡수선량이 0kGy, 5kGy, 10kGy, 30kGy, 50kGy이 되도록 하여 방사선 조사시 흡수선량에 따른 다당류의 추출Extraction of polysaccharides according to the absorbed dose at the time of irradiation by absorbing the radiation dose in the Example 2 to 0kGy, 5kGy, 10kGy, 30kGy, 50kGy
표 3. 해조류로부터 방사선 조사를 이용한 추출 공정으로 얻어진 다당류의 DPPH 라디칼 제어능Table 3. DPPH radical control ability of polysaccharides obtained by extraction process using radiation from seaweed
상기 표 3에서 방사선 조사 선량이 증가함에 따라 다당류의 DPPH 라디칼 제어능이 증가된 것을 확인하였다. In Table 3, as the radiation dose was increased, it was confirmed that the DPPH radical control ability of the polysaccharide was increased.
또한, 해조류로부터 감마선 조사 기술을 이용한 추출 방법으로 추출된 후코이단의 항산화력은 비조사 추출방법으로 추출된 후코이단에 비하여 50kGy 선량으로 감마선 조사한 추출 공정에서 얻어진 경우 약 12％ 증가하였으며, 감마선 조사 기술을 이용한 추출 방법으로 추출된 라미나린의 항산화력은 비조사 추출방법으로 추출된 라미나린에 비하여 50 kGy 선량으로 감마선 조사한 추출 공정에서 얻어진 경우 약 107％ 증가하였다.In addition, the antioxidant power of fucoidan extracted from seaweed by gamma-irradiation technique was increased by about 12% compared to fucoidan extracted by non-irradiation method, when it was obtained by gamma-irradiation at 50kGy dose. The antioxidant power of laminarin extracted by the extraction method was increased by about 107% when obtained by the gamma-irradiation extraction process with 50 kGy dose compared to laminarin extracted by the non-irradiation extraction method.
<시험예 4> 해조류 다당류의 항고혈압 활성 측정Test Example 4 Antihypertensive Activity of Seaweed Polysaccharides
방사선 조사를 이용하여 해조류로부터 추출된 다당류로서 상기 실시예 2에서 방사선 조사시 흡수선량이 0kGy, 5kGy, 10kGy, 30kGy, 50kGy이 되도록 하여 방사선 조사시 흡수선량에 따라 추출한 후코이단과 라미나린의 항고혈압 활성은 ACE(Angiotensin Converting Enzyme) 저해활성을 통해 측정하고 그 결과를 아래의 표 4에 나타내었다.Anti-hypertensive activity of fucoidan and laminarin extracted according to the absorbed dose at the time of irradiation with the polysaccharide extracted from the seaweed using the irradiation so that the absorbed dose is 0kGy, 5kGy, 10kGy, 30kGy, 50kGy when irradiated in Example 2 Was measured through ACE (Angiotensin Converting Enzyme) inhibitory activity and the results are shown in Table 4 below.
Hippury-L-Histidyl-L-Leusine(HHL) 25mg을 0.1M 소디움 보레이트 버 퍼(Sodium borate buffer, pH 8.3)에 용해하여 제조한 기질 30㎕에 시료로서 상기 실시예 2에서 방사선 조사시 흡수선량이 0kGy, 5kGy, 10kGy, 30kGy, 50kGy이 되도록 하여 방사선 조사시 흡수선량에 따라 추출한 후코이단과 라미나린을 각각 10㎕를 가하여 37℃에서 10분간 반응하였다. 그 다음 시료 대조군을 1N HCl 50㎕첨가하여 반응을 종료시킨 후 시료에 효소 ACE(0.5 unit/mL) 10㎕를 첨가하여 37℃에서 30분간 반응한 다음 1N HCl 50㎕첨가하여 반응을 종료하였다. 그후 모든 처리군에 에틸아세테이트(ethylacetate) 300㎕를 첨가한 다음 원심분리기를 이용하여 원심분리를 한 후 상층액 250㎕를 취해서 70℃에서 1시간 동안 방치하였다. 그 다음 증류수 300㎕를 첨가하여 교반 후 분광광도계(UV 1600 PC, Shimadzu, Tokyo, Japan)를 이용하여 228nm에서 흡광도를 측정하였다. 양성대조군으로는 Enalapril maleate salt(0.3 mg/ml)을 사용하였다. ACE 저해활성 환산식은 다음과 같다. Absorbed dose when irradiated in Example 2 as a sample to 30 μl of a substrate prepared by dissolving 25 mg of Hippury-L-Histidyl-L-Leusine (HHL) in 0.1 M sodium borate buffer (pH 8.3) 10 μl of fucoidan and laminarin were added to 0kGy, 5kGy, 10kGy, 30kGy, and 50kGy, and the amount of fucoidan and laminarin extracted according to the absorbed dose during irradiation was reacted at 37 ° C for 10 minutes. Then, the reaction was terminated by adding 50 μl of 1N HCl to the sample control group, and 10 μl of enzyme ACE (0.5 unit / mL) was added to the sample, followed by reaction at 37 ° C. for 30 minutes, and then 50 μl of 1N HCl was added to terminate the reaction. Thereafter, 300 μl of ethylacetate was added to all the treatment groups, followed by centrifugation using a centrifuge, and 250 μl of the supernatant was left at 70 ° C. for 1 hour. Then, 300 μl of distilled water was added thereto, and then the absorbance was measured at 228 nm using a spectrophotometer (UV 1600 PC, Shimadzu, Tokyo, Japan). Enalapril maleate salt (0.3 mg / ml) was used as a positive control. ACE inhibitory activity conversion formula is as follows.
ACE 저해활성 = 1 - {(S-S.C)/(B-B.C)×100} ACE inhibitory activity = 1-{(S-S.C) / (B-B.C) × 100}
S : 시료의 흡광도S: absorbance of the sample
S.C. : 시료 대조군의 흡광도S.C. : Absorbance of sample control
B : 공시험의 흡광도B: absorbance of blank test
B.C : 공시험 대조군의 흡광도B.C: absorbance of blank test control group
표 4. 해조류로부터 방사선 조사를 이용한 추출 공정으로 얻어진 기능성 다당류의 항고혈압능Table 4. Antihypertensive Capacity of Functional Polysaccharides Obtained by Extraction Process Using Irradiation from Seaweeds
표 4에서 해조류로부터 방사선 조사 기술을 이용한 추출공정으로 얻어진 다당류인 후코이단과 라미나린의 항고혈압능은 방사선 조사 선량이 증가함에 따라 다당류의 항고혈압능이 증가된 것을 확인하였다.In Table 4, the antihypertensive capacity of the polysaccharides fucoidan and laminarin obtained by the extraction process using the irradiation technique from the seaweeds was confirmed that the antihypertensive capacity of the polysaccharide increased as the irradiation dose increased.
<시험예 5> 방사선 조사를 이용하여 해조류로부터 추출된 다당류의 암세포에 대한 세포독성 효과 측정Test Example 5 Measurement of Cytotoxic Effects on Cancer Cells of Polysaccharides Extracted from Seaweed Using Irradiation
방사선 조사를 이용하여 해조류로부터 추출된 다당류로서 상기 실시예 2에서 방사선 조사시 흡수선량이 0kGy, 5kGy, 10kGy, 30kGy, 50kGy이 되도록 하여 방사선 조사시 흡수선량에 따라 추출한 후코이단의 암세포에 대한 세포독성 효과를 측정하였다.Cytotoxic effect on fucoidan cancer cells extracted according to the absorbed dose at the time of irradiation by the polysaccharide extracted from seaweed using radiation irradiation so that the absorbed dose is 0kGy, 5kGy, 10kGy, 30kGy, 50kGy when irradiated in Example 2 Was measured.
본 시험에 사용된 세포들은 B16BL6(피부암), AGS(위암), HT-29(결장암 ; colon cancer) 그리고 RAW264.7(마크로파지 세포)이며, 한국세포주은행(Korean Cell Line Bank)으로부터 분양받아 배양하면서 실험에 사용하였다. The cells used in this study were B16BL6 (Skin Cancer), AGS (Stomach Cancer), HT-29 (Colon Cancer; Colon Cancer) and RAW264.7 (Macrophage Cells), which were cultured by being distributed from the Korean Cell Line Bank. It was used for the experiment.
상기 B16BL6와 RAW264.7 세포주는 100 U/ml 페니실린과 100 U/ml 스트렙토마이신 및 10％ 소혈청(fetal bovine serum)이 함유된 EMEM 및 DMEM 배지를 사용하였고, AGS와 HT-29는 100U/ml 페니실린과 100U/ml 스트렙토마이신 및 10％ 소혈청이 함유된 RPMI 1640 배지를 사용하여 37℃, 5％ CO2 인큐베이터에서 배양하였다.The B16BL6 and RAW264.7 cell lines used EMEM and DMEM medium containing 100 U / ml penicillin, 100 U / ml streptomycin and 10% fetal bovine serum, and 100 U / ml of AGS and HT-29. Cultured in 37 ° C., 5% CO 2 incubator using RPMI 1640 medium containing penicillin, 100 U / ml streptomycin and 10% bovine serum.
방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단에 의한 암세포와 정상세포의 세포독성 평가는 MTT[3-(4,5-dimethylthiazolyl)-2,5-diphenyl-tetrazolium bromide, sigma] assay에 의해서 실험하였다. Evaluation of cytotoxicity of cancer cells and normal cells by fucoidan, a polysaccharide extracted from seaweed using irradiation, was investigated by MTT [3- (4,5-dimethylthiazolyl) -2,5-diphenyl-tetrazolium bromide, sigma] assay It was.
각각의 암세포와 정상세포는 96웰플레이트(well plate)에 웰당 3× 104cells/well의 농도로 분주하였다. 각각의 자숙액 시료는 농도 5mg/ml로 96 웰플레이트에 가하였다. 상기 플레이트를 37℃에서 24시간 배양한 후, 5mg/ml의 농도의 MTT 시약을 각각의 웰에 30㎕씩 가하였고, 37℃에서 2시간 동안 배양한 다음, 2000rpm에서 3분간 원심분리 하고, 상등액을 제거한 후, 100㎕의 DMSO(dimehylsulfoxide)를 첨가하고, 37℃에서 5분간 반응 후, 540nm에서 흡광도를 측정하였다.Each cancer cell and normal cells were divided into 96 well plates at a concentration of 3 × 10 4 cells / well per well. Each cooked liquid sample was added to a 96 well plate at a concentration of 5 mg / ml. After incubating the plate for 24 hours at 37 ° C, 30 μl of MTT reagent of 5 mg / ml was added to each well, incubated for 2 hours at 37 ° C, and then centrifuged at 2000 rpm for 3 minutes, and the supernatant. After the removal, 100 μl of DMSO (dimehylsulfoxide) was added, and after 5 minutes of reaction at 37 ° C., the absorbance was measured at 540 nm.
상기 암세포에 대한 항암효과를 측정하기 위하여, MTT 환원 검정법(MTT reduction assay)을 이용하여 방사선 조사하지 않은 대조구와 비교하여 암세포 성장억제 효과를 확인하였고 그 결과를 아래 표 5에 나타내었다. In order to measure the anticancer effect on the cancer cells, MTT reduction assay (MTT reduction assay) was used to confirm the cancer cell growth inhibitory effect compared to the control group not irradiated and the results are shown in Table 5 below.
방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단의 암세포 성장억제 효과를 측정한 결과 방사선 조사 선량이 증가할수록, HT-29의 세포독성은 34±1％ 까지 증가하였으며, B16BL6의 세포독성은 43±1％ 까지 증가하였다. 마지막으로 AGS의 세포독성은 82±1％ 까지 증가하였다. As a result of measuring the cancer cell growth inhibitory effect of fucoidan, a polysaccharide extracted from algae by irradiation, the cytotoxicity of HT-29 increased by 34 ± 1% and the cytotoxicity of B16BL6 by 43 ± Increased by 1%. Finally, the cytotoxicity of AGS increased to 82 ± 1%.
방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단의 암세포 성장억제 효과와 방사선이 조사되지 않은 해조류로부터 추출된 다당류인 후코이단 의 세포독성을 비교해 볼 때, 방사선 흡수선량이 5kGy, 10kGy, 30kGy, 50kGy이 되도록 방사선이 조사되어 해조류로부터 추출된 다당류인 후코이단은 방사선 흡수선량이 0kGy이 되도록 하여 방사선이 조사되지 않고 해조류로부터 추출된 다당류인 후코이단 보다 세포독성이 증가되었음을 알 수 있었다. 이러한 결과로부터 본 발명의 방사선 방사선 조사를 이용하여 해조류로부터 추출된 다당류가 암세포에 대한 저항성을 증가시켜준다는 것을 확인할 수 있었다.The radiation-absorbing doses of 5kGy, 10kGy, 30kGy, 50kGy were compared with the cancer cell growth inhibition effect of fucoidan, a polysaccharide extracted from seaweed, and the cytotoxicity of fucoidan, a polysaccharide extracted from seaweed without radiation. Fucoidan, a polysaccharide extracted from seaweed, was irradiated with radiation so that the radiation absorbed dose was 0 kGy, indicating that cytotoxicity was increased than fucoidan, a polysaccharide extracted from seaweed, without radiation. From these results, it was confirmed that the polysaccharide extracted from seaweed using the radiation irradiation of the present invention increases the resistance to cancer cells.
상기에서 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단의 암세포에 대한 세포독성이 방사선 조사선량이 증가함에 따라 증가함을 보였으며, 이러한 관점에서 정상세포에 대한 세포독성을 확인해 볼 필요가 있어, 상기 RAW264.7 세포주를 정상세포로 하여 정상세포 독성실험을 실시하였다. The cytotoxicity of cancer cells of fucoidan, a polysaccharide extracted from seaweed using radiation, was shown to increase with increasing radiation dose. From this point of view, it is necessary to confirm the cytotoxicity against normal cells. Normal cytotoxicity experiments were performed using the RAW264.7 cell line as normal cells.
방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단의 정상세포에 대한 세포독성 효과의 감소를 확인한 결과, 방사선 조사를 이용하여 해조류로부터 추출된 다당류는 방사선 조사선량이 증가함에 따라 5kGy에서는 세포독성이 24±1％까지 감소하였고, 10kGy에서는 세포독성이 17±1％까지 감소했으며, 50kGy에서의 세포독성은 3±1％까지 감소하였다. Irradiation confirmed the decrease of cytotoxic effect on the normal cells of fucoidan, a polysaccharide extracted from seaweeds. As a result, the polysaccharides extracted from seaweeds by irradiation increased the cytotoxicity at 5kGy as the radiation dose increased. Cytotoxicity was reduced by 17 ± 1% at 10 kGy and cytotoxicity was reduced by 3 ± 1% at 50 kGy.
표 5. 해조류로부터 방사선 조사를 이용한 추출 공정으로 얻어진 기능성 다당류의 암세포에 대한 세포 독성Table 5. Cytotoxicity of Cancer Cells of Functional Polysaccharides Obtained by Radiation Extraction from Seaweeds
본 실험을 통해 방사선 조사를 이용하여 해조류로부터 추출된 다당류인 후코이단은 방사선 조사를 하지 않고 해조류로부터 추출된 다당류에 대조해 볼 때, 정상세포에 대한 독성이 매우 적음을 알 수 있으며, 또한 방사선 조사를 이용하여 해조류로부터 추출된 다당류에 대한 암세포의 독성은 증가하지만, 정상세포에 대한 독성은 감소함을 알 수 있었다.This experiment shows that fucoidan, a polysaccharide extracted from seaweed using radiation, has little toxicity to normal cells when compared to polysaccharides extracted from seaweed without radiation. Thus, while the toxicity of cancer cells to polysaccharides extracted from seaweeds was increased, the toxicity to normal cells was reduced.
상술한 바와 같이, 본 발명의 바람직한 실시예를 참조하여 설명하였지만 해당 기술 분야의 숙련된 당업자라면 하기의 특허청구범위에 기재된 본 발명의 사상 및 영역으로부터 벗어나지 않는 범위 내에서 본 발명을 다양하게 수정 및 변경시킬 수 있음을 이해할 수 있을 것이다. As described above, although described with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and modified within the scope of the present invention without departing from the spirit and scope of the invention described in the claims below. It will be appreciated that it can be changed.
본 발명의 방사선 조사를 이용한 해조류로부터 다당류를 추출하는 방법은 방사선 조사량을 조절하여 해조류로부터 다당류를 고수율로 얻을 수 있어 종래 해조류로부터 다당류를 추출하는 방법에 의해 생산성을 증가시켜 제조비용을 감소시킬 수 있다.In the method of extracting polysaccharides from seaweeds using the radiation of the present invention, polysaccharides can be obtained in high yield by adjusting the radiation dose, and thus the production cost can be reduced by increasing the productivity by extracting polysaccharides from seaweeds. have.
또한 본 발명의 방사선 조사를 이용히여 해조류로부터 추출한 다당류는 항산화, 항고혈압능 및 암세포에 대한 세포독성의 기능성이 있어 이러한 다당류를 식품학적 조성물, 의약품 조성물 및/또는 화장품 조성물 등의 다양한 용도에 적용하여 소비자의 건강증진에 기여할 수 있다.In addition, the polysaccharide extracted from seaweed using the radiation of the present invention has the function of antioxidant, antihypertensive activity and cytotoxicity against cancer cells, so that the polysaccharide is applied to various uses such as food composition, pharmaceutical composition and / or cosmetic composition It can contribute to health promotion of consumers.
도 1은 해조류 세포의 Transmission Electron Microscope(TEM) 사진으로서 화살표는 세포벽을 나타내며, 도 1(a)는 방사선 조사를 하지 않은 해조류 세포의 TEM 사진이고, 도 1(b)는 10kGy 선량의 감마선을 조사한 해조류의 TEM 사진이고, 도 1(c)는 10kGy 선량의 전자선을 조사한 해조류의 TEM 사진이다.1 is a Transmission Electron Microscope (TEM) image of algae cells, an arrow indicates a cell wall, FIG. 1 (a) is a TEM image of an unirradiated algae cell, and FIG. 1 (b) shows gamma rays of 10 kGy dose. TEM picture of the seaweed, Figure 1 (c) is a TEM picture of the seaweed irradiated with an electron beam of 10kGy dose.
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|---|---|---|---|---|
| KR100623001B1 (en) * | 2004-10-11 | 2006-09-19 | 한국식품연구원 | A Method Process for Separation of Food Grade Laminaran and Fucoidan Having Properties of Anti-tumor and Anti-hyperlipidemic Polysaccharides from Brown algae |
| KR100679365B1 (en) | 2006-01-13 | 2007-02-06 | 박대환 | A manufacturing method of crude alginate |
| KR100760815B1 (en) | 2006-11-01 | 2007-10-04 | 오경덕 | A fermentation process for small molecular fucoidan from brown seaweed |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100623001B1 (en) * | 2004-10-11 | 2006-09-19 | 한국식품연구원 | A Method Process for Separation of Food Grade Laminaran and Fucoidan Having Properties of Anti-tumor and Anti-hyperlipidemic Polysaccharides from Brown algae |
| KR100679365B1 (en) | 2006-01-13 | 2007-02-06 | 박대환 | A manufacturing method of crude alginate |
| KR100760815B1 (en) | 2006-11-01 | 2007-10-04 | 오경덕 | A fermentation process for small molecular fucoidan from brown seaweed |
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