KR20200048049A - Antioxidant composition comprising extract of Apis mellifera male pupa - Google Patents
Antioxidant composition comprising extract of Apis mellifera male pupa Download PDFInfo
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- KR20200048049A KR20200048049A KR1020180129682A KR20180129682A KR20200048049A KR 20200048049 A KR20200048049 A KR 20200048049A KR 1020180129682 A KR1020180129682 A KR 1020180129682A KR 20180129682 A KR20180129682 A KR 20180129682A KR 20200048049 A KR20200048049 A KR 20200048049A
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- pupae
- extract
- western
- antioxidant
- alcohol
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
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- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
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Abstract
Description
본 발명은 서양종꿀벌(Apis mellifera)의 수번데기를 함유하는 항산화용 조성물에 관한 것이다. The present invention relates to an antioxidant composition containing several pupae of the Western honey bee ( Apis mellifera ).
오늘날 우리는 식량부족의 시대에 살고 있다. 이러한 세계의 식량 문제를 해결하기 위해 건강을 해치는 유전자 변형 농작물이 식탁을 점령하고, 많은 수확량을 위해 환경을 해치는 화학농약과 화학비료의 사용으로 환경오염을 유발하며 건강에도 많은 부작용을 발생시키고 있다. 또한, 에너지 문제, 가뭄, 홍수, 폭우 등의 기후 변화의 문제, 기후 변화와 관련된 온실가스의 문제 등으로 인해 세계의 생산곡물이 감소하고 있으며, 감소한 세계 생산곡물은 그 1/3이 가축의 먹이로 소비되고 있다. 아울러, 많은 가축은 메탄가스와 이산화탄소를 다량 발생시켜 지구온난화의 주범인 온실가스를 증가시키고 있어 심각한 환경 및 자원 위기에 직면하고 있다. 따라서, 축산물 소비를 최소화하고, 환경을 살리며, 양질의 대체식량자원을 개발할 필요성이 요구되었는바, 곤충은 육류에 비해 단백질 함유량이 평균 2배 이상 높고, 무기질, 필수 및 비필수 아미노산, 비타민 등이 풍부하며, 지방의 대부분이 수용성 지방으로 이루어져 있어 영양성분이 인체에 쉽게 흡수될 수 있어 축산물에서 얻는 단백질의 대체식품으로 각광받고 있다.Today we live in an era of food shortages. In order to solve the food problems in the world, genetically modified crops that harm health occupy the table, and use chemical pesticides and chemical fertilizers that harm the environment for large yields to cause environmental pollution and cause many side effects to health. In addition, the world's production crops are decreasing due to energy problems, droughts, floods, heavy rains, climate change problems, and climate change-related greenhouse gas problems. Is being consumed. In addition, many livestock face a serious environmental and resource crisis as they generate a large amount of methane gas and carbon dioxide to increase greenhouse gas, a major cause of global warming. Therefore, there is a need to minimize the consumption of livestock products, save the environment, and develop a high-quality alternative food resource. Insects have an average protein content of 2 times higher than meat, and mineral, essential and non-essential amino acids, vitamins, etc. It is abundant, and most of the fat is made of water-soluble fat, so nutrients can be easily absorbed by the human body.
우리나라는 2010년 곤충산업육성법 시행 후 곤충산업의 영역확대가 필요하나 현재까지 화분매개, 학습, 정서, 천적 및 환경 정화 관련 연구는 많이 진행되어왔으나 실제로 산업을 크게 확대할 수 있는 분야인 식품, 사료, 의약, 생체모방 및 생명공학 등의 연구는 아직 미흡한 실정이다.Although Korea needs to expand the scope of the insect industry after the Enforcement of the Insect Industry Promotion Act in 2010, there have been many studies related to pollen meditation, learning, emotion, natural enemies, and environment purification, but food and feed, which are fields that can greatly expand the industry , Pharmaceuticals, biomimetics, and biotechnology are still insufficient.
이러한 곤충 중에서, 지렁이, 지네, 백강잠, 굼벵이, 말벌집 등이 예로부터 약용으로 주로 많이 사용되었고, 식품으로서 벼메뚜기나 누에, 번데기 등이 식품으로 오래전부터 이용되어 왔다. 근래에 한시적 식품원료로서 갈색거저리와 흰점박이꽃무지의 유충과 귀뚜라미가 등록되었고, 이 중 갈색거저리와 귀뚜라미는 식품원료로 정식으로 인정받았다. 이 외의 다른 곤충들이 추가적인 식품원료로 이용되기 위해 각종 연구가 현재 활발히 진행 중이다. Among these insects, earthworms, centipedes, white ginseng, slugs, wasps, and the like have been mainly used for medicinal purposes, and rice locusts, silkworms, and pupae have been used as food for a long time. Recently, caterpillars and crickets of brown and white spotted flowers have been registered as temporary food ingredients, among which brown meal and crickets have been officially recognized as food ingredients. Various other researches are actively underway to use other insects as additional food ingredients.
이에 본 발명자들은 식품을 비롯하여 각종 약리학적 성분으로 이용가능한 곤충에 대해 연구하던 중, 양봉농가에서 불필요하여 버려지던 서양종꿀벌의 수번데기가 각종 영양성분이 풍부하면서도 항산화성이 있어 식품원료 또는 화장료 원료로 이용가능함을 확인하여 본 발명을 완성하게 되었다. Accordingly, the present inventors were studying insects that can be used as food and various pharmacological ingredients, and many of the honeybees that were thrown away by the bee farm are discarded because they are rich in various nutrients and have antioxidant properties. The present invention was completed by confirming the availability.
본 발명의 목적은 서양종꿀벌(Apis mellifera)의 수번데기를 함유하는 항산화용 조성물을 제공하는 데에 있다. An object of the present invention is to provide a composition for antioxidants containing several pupae of Western honey bee ( Apis mellifera ).
본 발명은 본 발명은 서양종꿀벌(Apis mellifera)의 수번데기 또는 이의 추출물을 함유하는 것을 특징으로 하는 항산화용 조성물에 관한 것이다. The present invention relates to a composition for antioxidant, characterized in that it contains several pupae or extracts of the Western honey bee ( Apis mellifera ).
상기 추출물은 서양종꿀벌 수번데기를 물, C1~C4 알코올 또는 이들의 혼합용액을 용매로 하여 추출하여 제조한 것일 수 있다. The extract may be prepared by extracting Western honeybee pupae using water, C1 to C4 alcohol, or a mixed solution thereof.
상기 C1~C4 알코올은 메탄올, 에탄올, 프로판올, 이소프로판올, 부탄올 및 이소부탄올로 이루어진 군에서 선택될 수 있다. The C1 ~ C4 alcohol may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.
상기 C1~C4 알코올은 특히 에탄올일 수 있다. The C1 ~ C4 alcohol may be particularly ethanol.
상기 추출에서 용매는 수번데기 사용 중량 기준 1~40배 부피를 사용할 수 있다. In the extraction, the solvent may use a volume of 1 to 40 times based on the weight of several pupae.
이에 본 발명에서는 서양종꿀벌(Apis mellifera)의 수번데기 또는 이의 추출물을 함유하는 것을 특징으로 하는 항산화용 건강기능식품 또는 화장료 조성물을 제공한다. Accordingly, the present invention provides a health functional food or cosmetic composition for antioxidant, characterized in that it contains several pupae or extracts of the Western honey bee ( Apis mellifera ).
또한 본 발명은 서양종꿀벌(Apis mellifera)의 수번데기를 세척 및 탈수하는 단계; 상기 수번데기를 건조한 후 분말화하는 단계; 및, 분말화된 수번데기에 물, C1~C4 알코올 또는 이들의 혼합용액을 가하여 추출한 후 건더기를 제거하여 얻은 여액을 농축 및 건조하는 단계;를 포함하는 것을 특징으로 하는 서양종꿀벌 수번데기 추출물의 제조방법을 제공한다. In addition, the present invention is a step of washing and dehydrating the pupae of the Western honey bee ( Apis mellifera ); Drying the pupae and pulverizing it; And extracting by adding water, C1 to C4 alcohol or a mixed solution thereof to the powdered pupae, and then concentrating and drying the filtrate obtained by removing the dried crust; production of Western honeybee pupae pupa extract comprising Provide a method.
이하, 본 발명을 상세하게 설명한다.Hereinafter, the present invention will be described in detail.
상기 서양종꿀벌 수번데기의 추출물은 수번데기를 물, C1~C4 알코올 또는 이들의 혼합용액을 용매로 하여 추출할 수 있으며, 상기 C1~C4 알코올은 메탄올, 에탄올, 프로판올, 이소프로판올, 부탄올 및 이소부탄올로 이루어진 군에서 선택될 수 있다. The extract of the Western honey bee pupae can extract the pupae using water, C1 ~ C4 alcohol or a mixed solution thereof as a solvent, and the C1 ~ C4 alcohol is methanol, ethanol, propanol, isopropanol, butanol and isobutanol It can be selected from the group consisting.
상기 서양종꿀벌 수번데기의 추출물의 제조시 사용되는 물, C1~C4 알코올 또는 이들의 혼합용액은 서양종꿀벌 수번데기 사용 중량 기준 1~40배 부피(1kg 기준 1~40ℓ)를 사용할 수 있으며, 바람직하게는 5~40배 부피를 사용할 수 있다. 상기 서양종꿀벌 수번데기 추출물의 추출조건은 20~100℃에서 1분~48시간일 수 있다. 상기 과정은 1~4번까지 반복할 수 있다. Water, C1 ~ C4 alcohol or a mixed solution used in the preparation of the extract of the Western honey bee pupae may use 1 to 40 times the volume (1 to 40 l based on 1 kg) by weight using the Western bee pupae, preferably Can use 5 to 40 times the volume. The extraction conditions of the Western honey bee pupa extract may be from 1 minute to 48 hours at 20 to 100 ° C. The above process can be repeated 1 to 4 times.
또한, 당분야의 통상적인 방법으로서 상기 서양종꿀벌 수번데기의 물, C1~C4 알코올 또는 이들의 혼합용액 추출물을 물에 녹인 후에 n-헥산, 클로로포름, 에틸아세테이트 및 n-부탄올로 이루어진 군 중에서 선택되는 1종 이상의 용매를 사용하여 추가적으로 분획하여 분획물로 제조할 수 있다. In addition, as a conventional method in the art, after dissolving water, C1 ~ C4 alcohol or a mixed solution extract of the Western honeybee pupae in water, it is selected from the group consisting of n-hexane, chloroform, ethyl acetate and n-butanol. It can be prepared as a fraction by further fractionation using one or more solvents.
또다른 방법으로는, 서양종꿀벌 수번데기를 물, C1~C4 알코올 또는 이들의 혼합용매로 추출 농축하여 얻은 서양종꿀벌 수번데기 추출물에, 물을 가하여 현탁한 후, 바람직하게는 서양종꿀벌 수번데기 추출물의 중량의 1~1000배, 더 바람직하게는 1~500배, 가장 바람직하게는 1~50배의 물을 가하여 현탁한 후, 상기 현탁물에 n-헥산, 클로로포름, 에틸아세테이트 및 n-부탄올로 이루어진 군에서 선택되는 용매를 가하여 얻은 서양종꿀벌 수번데기 분획물로 제조할 수 있다. 상기 서양종꿀벌 수번데기 분획물은 바람직하게는, 서양종꿀벌 수번데기를 물, C1~C4 알코올 또는 이들의 혼합용매로 추출 농축하여 얻은 서양종꿀벌 수번데기 추출물을 물에 현탁한 후 헥산과 혼합하여 얻은 헥산층의 농축물, 상기 헥산층을 제거하고 남은 잔사(물층)에 클로로포름을 혼합하여 얻은 클로로포름층의 농축물, 또는 상기 클로로포름층을 제거하고 남은 잔사(물층)에 에틸아세테이트를 혼합하여 얻은 에틸아세테이트층의 농축물, 상기 에틸아세테이트층을 제거하고 남은 잔사(물층)에 부탄올을 혼합하여 얻은 부탄올층의 농축물, 또는 상기 부탄올층을 제거하고 남은 잔사(물층)의 농축물일 수 있다. 한편, 이 외의 분획조건은 제한되지는 않으나, 상기 서양종꿀벌 수번데기 추출물에 에리트로플레움 포르디 추출물의 중량의 1~50배의 물을 가하여 현탁물을 제조한 후, 상기 물과 동량의 헥산, 클로로포름, 에틸아세테이트, 및, 부탄올로 이루어진 군에서 선택되는 용매를 가하여 분획할 수 있다. 또한, 헥산층을 제거한 후 남은 잔사에 클로로포름을 가하고, 클로로포름층을 제거하고 남은 잔사에 에틸아세테이트를 가하고, 에틸아세테이트를 제거하고 남은 잔사에 부탄올을 가할 때에도, 단계적으로 이루어 질 때도 역시 잔사와 동량의 각 용매(클로로포름, 에틸아세테이트 또는 부탄올)를 순차적으로 가하여 분획할 수 있다. In another method, after suspending Western honeybee pupae with water, C1 ~ C4 alcohol or a mixed solvent thereof, and then suspending the Western honeybee pupae extract obtained by adding water, preferably, the Western honeybee pupae extract After 1 to 1000 times the weight, more preferably 1 to 500 times, most preferably 1 to 50 times water, and suspended, the suspension consists of n-hexane, chloroform, ethyl acetate and n-butanol. It can be prepared from fractions of Western honeybee pupa obtained by adding a solvent selected from the group. The Western honey bee pupa fraction is preferably a hexane layer obtained by suspending the Western honey bee pupa pupa extract obtained by extracting and concentrating the Western honey bee pupae with water, C1 to C4 alcohol or a mixed solvent thereof, and then mixing it with hexane. The concentrate of the chloroform layer obtained by removing the hexane layer and mixing chloroform with the remaining residue (water layer), or the ethyl acetate layer obtained by removing the chloroform layer and mixing ethyl acetate with the remaining residue (water layer) The concentrate may be a concentrate of a butanol layer obtained by mixing butanol with the residue (water layer) after removing the ethyl acetate layer, or a concentrate of the residue (water layer) remaining after removing the butanol layer. On the other hand, the other fractional conditions are not limited, but after preparing a suspension by adding 1 to 50 times the weight of the weight of the erythropolium pordi extract to the Western honeybee pupa extract, hexane in the same amount as the water, It can be fractionated by adding a solvent selected from the group consisting of chloroform, ethyl acetate, and butanol. Also, after removing the hexane layer, chloroform is added to the remaining residue, chloroform layer is removed, ethyl acetate is added to the remaining residue, and ethyl acetate is removed and butanol is added to the remaining residue. Each solvent (chloroform, ethyl acetate or butanol) can be added sequentially and fractionated.
상기 추출물 또는 이의 분획물의 제조온도는 20 내지 100℃일 수 있으나, 이에 제한되는 것은 아니다. 추출 또는 분획 시간은 특별히 제한되는 것은 아니나, 10분 내지 2일 이내에 추출하는 것이 바람직하며, 추출용 기기로는 통상의 추출기기, 초음파분쇄추출기 또는 분획기를 이용할 수 있다. 이렇게 제조된 서양종꿀벌 수번데기 추출물 또는 분획물은 열풍건조, 감압건조 또는 동결건조하여 용매를 제거할 수 있다. 또한, 상기 서양종꿀벌 수번데기 추출물 또는 분획물은 칼럼크로마토그래피를 이용하여 정제하여 사용할 수 있다. The preparation temperature of the extract or its fraction may be 20 to 100 ° C, but is not limited thereto. The extraction or fractionation time is not particularly limited, but it is preferable to extract within 10 minutes to 2 days, and an ordinary extraction device, an ultrasonic pulverization extractor, or a fractionator may be used as the extraction device. The thus-prepared Western honey bee pupa extract or fraction can be removed by hot air drying, vacuum drying or freeze drying. In addition, the Western honey bee pupa extract or fraction can be purified by column chromatography and used.
상기 서양종꿀벌 수번데기 추출물은 상법에 따라, 유기용매(알코올, 에테르, 아세톤 등)에 의한 추출, 헥산과 물의 분배, 칼럼크로마토그래피에 의한 방법 등, 식물체 성분의 분리 추출에 이용되는 공지의 방법을 단독 또는 적합하게 조합한 방법을 이용하여 분획 또는 정제하여 사용할 수 있다. The Western honey bee pupae extract is a known method used for separation and extraction of plant components, such as extraction with an organic solvent (alcohol, ether, acetone, etc.), distribution of hexane and water, and method by column chromatography, according to the commercial method. It can be used by fractionation or purification using a single or appropriately combined method.
상기 크로마토그래피는 실리카겔 컬럼 크로마토그래피(silica gel column chromatography), 엘에이취-20 컬럼 크로마토그래피(LH-20 column chromatography), 이온교환수지 크로마토그래피(ion exchange resin chromatography), 중압 액체 크로마토그래피(medium pressure liquid chromatography), 박층 크로마토그래피(TLC; thin layer chromatography), 실리카겔 진공 액체 크로마토그래피(silica gel vacuum liquid chromatography) 및 고성능 액체 크로마토그래피(high performance liquid chromatography) 중에서 선택될 수 있다. The chromatography is silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.
또한, 본 발명은 서양종꿀벌 수번데기 또는 이의 추출물을 함유하는 항산화용 약학조성물을 제공한다. 상기 서양종꿀벌 수번데기 또는 이의 추출물은 본 발명의 약학 조성물에 0.001~100 중량%로 하여 첨가될 수 있다.In addition, the present invention provides a pharmaceutical composition for antioxidant that contains Western honeybee pupae or an extract thereof. The Western honey bee pupae or its extract may be added to the pharmaceutical composition of the present invention as 0.001 to 100% by weight.
상기 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 상기 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 본 발명의 추출물에 적어도 하나 이상의 부형제, 예를 들면, 전분, 탄산칼슘, 수크로스 또는 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. The pharmaceutical composition may be formulated and used in the form of oral dosage forms, external preparations, suppositories, and sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., according to a conventional method. Carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the extract of the present invention, for example, starch, calcium carbonate, sucrose or lactose, It is prepared by mixing gelatin. In addition, lubricants such as magnesium stearate and talc are used in addition to simple excipients. Liquid preparations for oral use include suspensions, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.
본 발명의 약학 조성물의 투여량은 치료받을 대상의 연령, 성별, 체중과, 치료할 특정 질환 또는 병리 상태, 질환 또는 병리 상태의 심각도, 투여경로 및 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있으며, 일반적으로 투여량은 0.01㎎/㎏/일 내지 대략 2000㎎/㎏/일의 범위이다. 더 바람직한 투여량은 1㎎/㎏/일 내지 500㎎/㎏/일이다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The dosage of the pharmaceutical composition of the present invention will vary depending on the age, gender, weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration, and the judgment of the prescriber. Dosage determination based on these factors is within the level of those skilled in the art, and dosages generally range from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be administered once a day, or may be divided into several times. The above dosage does not limit the scope of the present invention in any way.
본 발명의 약학 조성물은 쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내 주사에 의해 투여될 수 있다. 본 발명의 추출물은 독성 및 부작용이 거의 없으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있는 약제이다. The pharmaceutical composition of the present invention can be administered to various mammals, such as rats, livestock, and humans. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracranial injection. Since the extract of the present invention has little toxicity and side effects, it is a drug that can be used safely even for a long period of time for prevention purposes.
또한, 본 발명은 서양종꿀벌 수번데기 또는 이의 추출물 및 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 항산화용 건강기능식품을 제공한다. 상기 서양종꿀벌 수번데기 또는 이의 추출물은 본 발명의 건강기능식품에 0.001~100 중량%로 하여 첨가될 수 있다. 본 발명의 건강기능식품은 정제, 캡슐제, 환제 또는 액제 등의 형태를 포함하며, 본 발명의 서양종꿀벌 수번데기 또는 이의 추출물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제 등이 있다. In addition, the present invention provides a health functional food for antioxidant, including Western honey bee pupae or an extract thereof and a food-acceptable food supplement additive. The Western honey bee pupae or its extract can be added to the health functional food of the present invention at 0.001 to 100% by weight. The health functional food of the present invention includes tablets, capsules, pills or liquids, etc., and foods to which the Western honeybee pupae or extracts of the present invention can be added include, for example, various drinks, meat, Sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, beverages, alcoholic beverages, chewing gum, tea and vitamin complexes.
또한, 본 발명은 서양종꿀벌 수번데기 또는 이의 추출물을 포함하는 항산화용 화장료 조성물을 제공한다. 상기 화장료 조성물의 제형은 바람직하게는 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐로션, 영양로션, 맛사지크림, 영양크림, 모이스쳐크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클렌저로 이루어진 그룹에서 선택될 수 있다. 상기 화장료 제형에는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고 지질 및 해초 엑기스로 이루어진 군에서 선택된 성분이 추가로 포함될 수 있으며, 통상의 화장료에 포함되는 각종 부형제가 더 첨가될 수 있다.In addition, the present invention provides a cosmetic composition for antioxidant, including Western honeybee pupae or extracts thereof. The formulation of the cosmetic composition is preferably a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, It can be selected from the group consisting of pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser. The cosmetic formulation may further include components selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingoli lipids, and seaweed extracts, and various excipients included in conventional cosmetics may be further added.
본 발명은 서양종꿀벌(Apis mellifera)의 수번데기를 함유하는 것을 특징으로 하는 항산화용 조성물에 관한 것으로서, 상기 서양종꿀벌 수번데기 추출물이 포함된 조성물은 DPPH의 억제효과가 우수하여 체내 생리활성을 증진시키는 각종 약학 조성물과 건강기능식품으로서 용이하게 이용될 수 있다.The present invention relates to an antioxidant composition characterized in that it contains several pupae of the Western honey bee ( Apis mellifera ), and the composition containing the Western honey bee pupae extract has excellent inhibitory effect of DPPH to enhance the physiological activity in the body. It can be easily used as various pharmaceutical compositions and health functional foods.
도 1은 서양종꿀벌(Apis mellifera)의 수번데기(수벌 번데기), 수벌유충, 일벌유충, 일번번데기 각각의 에탄올 추출물이 갖는 DPPH 억제 활성을 확인한 결과 그래프이다. 1 is a graph showing the results of confirming the DPPH inhibitory activity of ethanol extracts of a number of pupae (a pupa pupae), a pupa larvae, a bee larvae and a pupae of the Western honey bee ( Apis mellifera ).
이하 본 발명의 바람직한 실시예를 상세히 설명하기로 한다. 그러나, 본 발명은 여기서 설명되는 실시예에 한정되지 않고 다른 형태로 구체화될 수도 있다. 오히려, 여기서 소개되는 내용이 철저하고 완전해지도록, 당업자에게 본 발명의 사상을 충분히 전달하기 위해 제공하는 것이다. Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided to sufficiently convey the spirit of the present invention to those skilled in the art so that the contents introduced herein are thorough and complete.
<실시예 1. 서양종꿀벌 수번데기의 수집><Example 1. Collection of Western honeybee pupae>
서양종꿀벌(Apis mellifera) 사양농가의 벌통에서 생산한 21~24일령의 수번데기를 채취하여 성분분석에 사용하였다. 수번데기는 채취 즉시 -20℃ 냉동고에 보관하였다.Western honey bee ( Apis mellifera ) The 21-24-day-old pupa produced from the beehive of a speculative farm was collected and used for ingredient analysis. Several pupae were immediately stored in a freezer at -20 ℃.
<실시예 2. 서양종꿀벌 수번데기의 성분분석><Example 2. Component analysis of Western honeybee pupae>
실시예 2-1. 일반성분 분석Example 2-1. General component analysis
수번데기의 일반성분 분석은 식품의약품안전처의 식품공전의 식품성분시험법에 따라 실시하였다. 즉, 수분함량은 105℃ 건조기를 이용하여 상압가열건조법에 의하여 측정하였고 회분함량은 검체가 백색~회백색의 회분이 얻어질 때까지 550~600℃에서 가열하는 직접회분법을 이용하였다. 조단백질 함량은 Kjeldahl법을 이용하여 검체에 단백질 분해촉진제와 황산을 넣어 분해한 다음 FOSS kjeltec 8400(Fisher Scientific, Hampton, NH, USA) 단백질 자동분석기로 함량을 측정하였다. 조지방 함량분석은 석유에테르를 추출용매로 Soxhlet법을 이용하였다. 탄수화물 함량은 검체 100g 중에 조단백질, 조지방, 수분 및 회분함량 뺀 나머지 값으로 나타내었다.The analysis of the general composition of several pupae was conducted in accordance with the Food Ingredient Test Method of the Food Code of the Ministry of Food and Drug Safety. That is, the moisture content was measured by a normal pressure heating and drying method using a 105 ° C dryer, and the ash content was a direct batch method in which samples were heated at 550 to 600 ° C until white to grayish white ash was obtained. The crude protein content was decomposed by adding a proteolytic accelerator and sulfuric acid to the sample using the Kjeldahl method, and then the content was measured by an FOSS kjeltec 8400 (Fisher Scientific, Hampton, NH, USA) protein automatic analyzer. In the analysis of crude fat content, Soxhlet method was used as an extraction solvent for petroleum ether. The carbohydrate content was expressed as the remaining value minus crude protein, crude fat, moisture and ash content in 100 g of the sample.
한편, 실시예 1에서 냉동된 시료를 분석에 사용하되 수번데기의 구성성분 변화를 고려하여 건조를 하지 않고 냉동시료를 녹인 후 균질화하여 분석에 사용하였다. 수번데기에 존재하는 탄수화물, 조지방, 조단백질의 3대 영양소를 비롯하여 회분 및 수분에 대한 분석결과를 표 1에 나타내었다. 수번데기의 회분은 0.85%, 수분함량은 74.23%로 가장 많은 함유량을 보였으며, 생물체의 영양에 가장 중요한 탄수화물, 지방, 단백질 각각 5.6%, 8.19%, 11.05%로, 단백질 함량이 가장 높게 나타났다. 결과적으로 수번데기는 식품으로서 요구되는 3대 영양소를 고르게 분포하고 있으며 특히 단백질 함량이 높아 환자들의 체력유지 및 회복에 도움이 되는 건강기능식품으로의 이용이 가능함을 확인할 수 있었다. On the other hand, the frozen sample in Example 1 was used for the analysis, but it was homogenized after melting and freezing the frozen sample without drying considering the change in the composition of several pupae. Table 1 shows the analysis results for ash and moisture, including three major nutrients of carbohydrate, crude fat, and crude protein present in several pupae. The ash content of male pupae was 0.85% and the moisture content was 74.23%, which was the most important. Carbohydrates, fats, and proteins, which are the most important for the nutrition of organisms, were 5.6%, 8.19%, and 11.05%, respectively, with the highest protein content. As a result, it was confirmed that several pupae are distributed evenly among the three major nutrients required as food, and especially, the protein content is high, so it can be used as a health functional food that helps patients maintain and recover their fitness.
실시예 2-2. 아미노산 조성 및 함량분석Example 2-2. Amino acid composition and content analysis
아미노산은 체내 단백질의 기본 구성단위이자 각종 신진대사에 촉매제로 작용하며 인체 조직의 재생과 회복에 관여한다(Sakami and Harrington, 1963). 필수아미노산은 체내에서 합성이 되지 않는 아미노산으로 식품단백질의 영양적 가치를 평가하는데 가장 중요한 척도로 알려져 있다(Sakami and Harrington, 1963). Amino acids are the basic building blocks of proteins in the body and act as catalysts for various metabolism and are involved in the regeneration and recovery of human tissues (Sakami and Harrington, 1963). Essential amino acids are amino acids that are not synthesized in the body and are known to be the most important measure of the nutritional value of food proteins (Sakami and Harrington, 1963).
건강기능식품공전의 건강기능식품 시험법에 따라 알라닌 등 16종의 아미노산은 2-머캅토에탄올(C2H6OS)을 함유한 6 N 염산을 사용하여 검체의 단백질을 분해한 다음, 염산을 제거하고 구연산나트륨 완충액을 첨가하여 여과한 후 ion exchange column이 장착된 아미노산 자동분석기(Hitachi L-8900, Tokyo, Japan)를 이용하여 16종의 아미노산을 동시분석하였다. According to the health functional food test method before the health functional food, 16 kinds of amino acids such as alanine were decomposed protein in the sample using 6 N hydrochloric acid containing 2- mercaptoethanol (C 2 H 6 OS), and then hydrochloric acid was used. After removal and filtering by adding sodium citrate buffer, 16 amino acids were simultaneously analyzed using an amino acid analyzer equipped with an ion exchange column (Hitachi L-8900, Tokyo, Japan).
시스테인 분석은 검체에 개미산을 첨가하고 하루정도 방치한 후 물을 추가하여 동결건조한 다음 2-머캅토에탄올을 함유한 6 N 염산을 사용하여 가수분해한 후 유도체화하여 octadecyl silica column이 장착된 HPLC를 이용하여 분석하였다. 이동상은 (A)40 mM NaH2PO4와 (B)MeCN:MeOH:H2O=45:45:10 용액을 (A)95~44%;0-31분, (A)44%;31~33분의 기울기 조건으로 설정하였고, 검출파장 Ex 262nm, Em 338nm, 컬럼온도 40℃, 흐름속도 1.5mL/min로 분석조건을 설정하였다. For cysteine analysis, formic acid was added to the sample, and after standing for about a day, water was added and lyophilized, followed by hydrolysis using 6N hydrochloric acid containing 2-mercaptoethanol and derivatization to obtain HPLC with octadecyl silica column. Analysis. Mobile phase (A) 40 mM NaH 2 PO 4 and (B) MeCN: MeOH: H 2 O = 45: 45: 10 solution (A) 95-44%; 0-31 min, (A) 44%; 31 It was set to the gradient condition of ∼33 minutes, and the analysis conditions were set to detection wavelength Ex 262nm, Em 338nm,
트립토판은 검체에 가용성전분과 수산화나트륨 용액을 가하고 동결시킨 후 탈기하고 밀봉하여 135℃에서 22시간동안 가수분해하였다. 가수분해물은 염산으로 중화시킨 후 원심분리하여 상층액을 분석시료로 사용하였으며 아미노산 자동분석기를 이용하여 함량을 측정하였다.Tryptophan was added to a sample of soluble starch and sodium hydroxide solution, frozen, degassed, sealed and hydrolyzed at 135 ° C for 22 hours. The hydrolyzate was neutralized with hydrochloric acid and centrifuged to use the supernatant as an analysis sample and the content was measured using an automatic amino acid analyzer.
표 2를 참고하면, 서양종꿀벌 수번데기의 아미노산 조성 분석결과, 18종의 아미노산이 검출되었으며 뇌의 기능을 활성화시키는 글루탐산(Gollnitz and Wiechert, 1967)의 경우 1631.9mg/100g으로 가장 높은 함량을 보였다. Referring to Table 2, as a result of the amino acid composition analysis of the Western honeybee pupae, 18 amino acids were detected and the highest content was 1631.9mg / 100g for glutamic acid (Gollnitz and Wiechert, 1967), which activates brain function.
그 다음으로 체내 암모니아 배출 및 간보호 기능을 가지는 동시에 질소대사와 에너지대사에 관여하여 피로저항력을 높여주는 아스파르트산(Sakami and Harrington, 1963)이 989.4mg/100g으로 확인되었으며 나머지 17종의 아미노산은 17.3~731.5mg/100g으로 측정되었다. Next, 989.4 mg / 100g of aspartic acid (Sakami and Harrington, 1963), which has a function of ammonia discharge and liver protection in the body and increases fatigue resistance by being involved in nitrogen metabolism and energy metabolism, was found to be 17.3. It was measured as ~ 731.5mg / 100g.
수번데기에는 발린, 루신, 이소루신, 페닐알라닌, 트립토판, 메티오닌, 트레오닌, 라이신, 히스티딘 등 9종의 필수아미노산이 포함되어 있으며 근육의 에너지원으로 이용되는 루신(Dohm et al., 1985)이 731.5mg/100g으로 가장 많은 함유량을 나타내었다. 따라서 수번데기에 19종의 아미노산이 함유되어 있어 양질의 단백질을 갖는 식품으로의 이용가능성이 충분함을 확인할 수 있다. The pupae contains nine essential amino acids such as valine, leucine, isoleucine, phenylalanine, tryptophan, methionine, threonine, lysine, and histidine, and 731.5 mg of leucine (Dohm et al ., 1985) used as a muscle energy source. / 100g showed the most content. Therefore, it can be confirmed that the availability of foods having high-quality protein is sufficient because 19 kinds of amino acids are contained in several pupae.
실시예 2-3. 무기물 조성 및 함량분석Example 2-3. Mineral composition and content analysis
무기물은 필수아미노산 같이 체내에서 합성되지 않은 물질로 생체기능의 유지를 위해서 필요한 미량 영양소로 분류되며 신체의 골격 형성, 삼투압의 조절, 근육의 기능 유지, 에너지 대사 등 다양한 기능이 있는 것으로 알려져 있다(Blumfield et al., 2013). Minerals are substances that are not synthesized in the body, such as essential amino acids, and are classified as micronutrients necessary for the maintenance of biological functions, and are known to have various functions such as the formation of the skeleton of the body, regulation of osmotic pressure, maintenance of muscle functions, and energy metabolism (Blumfield et al., 2013).
무기질 중 나트륨(Na), 마그네슘(Mg), 칼륨(K), 칼슘(Ca), 망간(Mn), 철(Fe), 구리(Cu), 아연(Zn), 인(P)은 식품공전의 일반시험법과 건강기능식품공전의 건강기능식품 시험법에 제시된 건식분해법에 따라 검체를 건조하여 탄화시키고 450~550℃에서 완전히 회화시킨 후 염산을 추가하여 유리여과기로 여과한 다음 유도 결합 플라즈마 발광분석기(ICAP 7400 Duo, Thermo Fisher Scientific, Waltham, MA, USA)를 이용하여 함량을 측정하였다. 몰리브덴(Mo) 및 셀레늄(Se) 역시 건식분해법에 의하여 시료를 전처리하고 함량분석은 Agilent ICP-MS 7700(Santa Clara, CA, USA)을 사용하였다. 염소(Cl) 분석은 식염 1g을 함유하는 양의 검체를 회화시킨 후 물에 녹이고 여과하여 크롬산칼륨시액을 첨가한 다음 질산은 용액으로 적정하여 염소의 양을 산출하였다.Among the minerals, sodium (Na), magnesium (Mg), potassium (K), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), and phosphorus (P) After drying and carbonizing the sample according to the dry decomposition method presented in the General Test Method and the Health Functional Food Test Method before the Health Functional Food Test, and completely painting at 450 ~ 550 ℃, add hydrochloric acid, filter with a glass filter, and then conduct an inductively coupled plasma luminescence analyzer ( ICAP 7400 Duo, Thermo Fisher Scientific, Waltham, MA, USA). Molybdenum (Mo) and selenium (Se) were also pre-treated by dry decomposition, and the content analysis was performed using Agilent ICP-MS 7700 (Santa Clara, CA, USA). In the analysis of chlorine (Cl), a sample containing an amount of 1 g of salt was painted, dissolved in water, filtered and potassium chromate solution was added, followed by titration with silver nitrate solution to calculate the amount of chlorine.
수번데기에 존재하는 무기물은 표 3의 결과와 같이, 12종의 무기물이 존재하는 것으로 나타났으며, 에너지 대사에 관여하는 칼륨과 인은 각각 235.78mg/100g, 177.73mg/100g으로 10종의 다른 무기물보다 4배 이상 높은 것으로 확인되었다. As shown in Table 3, the minerals present in several pupae were found to have 12 kinds of minerals. Potassium and phosphorus involved in energy metabolism were 235.78mg / 100g and 177.73mg / 100g, respectively. It was found to be four times higher than the mineral.
서양종 꿀벌의 먹이인 꿀과 화분은 일벌이 밀원식물에서 채취하여 벌집에 저장한 물질로 아미노산과 각종 무기물을 함유하는 것으로 보고되어 있는데(김 등, 2017; 홍 등, 2017), 무기물이 체내에서 합성되지 않는 점으로 볼 때, 수벌유충이 성충이 되기 전까지 섭취하는 꿀과 화분으로부터 무기물이 체내에 축적되어 수번데기에 존재하는 것으로 파악된다.Honey and pollen, which are food for Western-style bees, have been reported to contain amino acids and various minerals as a material collected by honeybees from wheat plants and stored in honeycombs (Kim et al., 2017; Hong et al., 2017). It is understood that minerals are accumulated in the body from honey and pollen consumed until the bee larvae become adults, and are present in several pupae.
실시예 2-4. 비타민 조성 및 함량분석Example 2-4. Vitamin composition and content analysis
비타민은 체내에서 생리 기능의 조절 및 물질대사에 관여하는 물질로 적응 양으로도 생리활성을 나타내는 필수적인 영양소이며 지용성과 수용성 비타민으로 분류한다(Khadim, 1981). Vitamins are substances that are involved in the regulation of physiological functions and metabolism in the body, and are essential nutrients that exhibit physiological activity even in adaptive amounts and are classified as fat-soluble and water-soluble vitamins (Khadim, 1981).
건강식품공전의 비타민 시험법에 따라 시료의 전처리 및 함량분석을 실시하였다. 비타민 A는 검체에 에탄올, 피로갈롤에탄올 및 수산화칼륨 용액을 넣고 비누화 시킨 다음 물과 석유에테르를 가하여 분획한 후 석유에테르층을 취하여 감압농축하고 이소프로판올을 넣어 분석시료로 사용하였다. 비타민 B1은 검체에 삼염화초산용액을 넣고 균질화한 다음 원심분리하여 상층액을 취한 후 초산나트륨용액과 카디아스타제 용액을 첨가한 다음 37℃에서 10시간 방치하여 시험용액으로 사용하였다. 비타민 B12는 검체에 인산완충용액을 가하고 초음파로 추출한 후 21,000rpm에서 원심분리를 실시하여 침전물을 제외한 하층에 클로로포름을 첨가하여 지방층을 제거하고 남은 물층을 멤브레인 필터로 여과하여 분석에 사용하였다. Pre-treatment and content analysis of samples were conducted according to the vitamin test method of the Health Food Code. Vitamin A was saponified by adding a solution of ethanol, pyrogallol ethanol, and potassium hydroxide to the sample, fractionated by adding water and petroleum ether, and then the petroleum ether layer was concentrated under reduced pressure and isopropanol was used as an analysis sample. Vitamin B 1 was used as a test solution by adding triacetic acid solution to the sample, homogenizing it, centrifuging it, taking the supernatant, adding sodium acetate solution and cardiatase solution, and then leaving it at 37 ° C for 10 hours. Vitamin B 12 was added to the sample with a phosphate buffer solution, extracted with ultrasound, and centrifuged at 21,000 rpm to remove the fat layer by adding chloroform to the lower layer excluding the precipitate, and filter the remaining water layer with a membrane filter for analysis.
비타민 C는 검체에 10% 메탄인산용액을 첨가하여 균질화한 후 3000rpm에서 원심분리한 다음 상층액을 시험용액으로 사용하였다. 비타민 D2와 D3는 검체에 피로갈롤에탄올을 가하여 진탕 혼합하고 90% 수산화칼륨을 넣어 비누화 한 다음 헥산을 넣어 분획을 실시하였다. 헥산층을 취하고 감압농축한 다음 메탄올을 가하여 멤브레인 필터로 여과하여 시험용액으로 사용하였다. Vitamin C was homogenized by adding 10% methane phosphate solution to the sample, centrifuged at 3000 rpm, and the supernatant was used as a test solution. Vitamin D 2 and D 3 were added to pyrogallol ethanol to the sample, shake-mixed, saponified with 90% potassium hydroxide, and fractionated with hexane. The hexane layer was taken, concentrated under reduced pressure, and then methanol was added and filtered through a membrane filter to be used as a test solution.
비타민 E는 검체에 헥산을 넣고 추출한 다음 여과하여 분석에 사용하였으며 α-tocopherol의 표준 검량선으로부터 비타민 E의 함량을 계산하였다. 비타민7종에 대한 정량분석은 HPLC(Shiseido Nanospace SI-2, Tokyo, Japan)를 사용하였으며 컬럼, 컬럼오븐온도, 흐름속도, 주입량, 이동상 등의 분석조건은 건강기능식품공전 일반시험법 비타민 분석법과 동일하였다. Vitamin E was extracted with hexane in the sample, filtered, and used for analysis. The content of vitamin E was calculated from the standard calibration curve of α -tocopherol. Quantitative analysis of 7 kinds of vitamins was carried out using HPLC (Shiseido Nanospace SI-2, Tokyo, Japan) .The analysis conditions such as column, column oven temperature, flow rate, injection amount, mobile phase, etc. It was the same.
표 4를 참고하면, 수번데기에 존재하는 비타민은 수용성 비타민 B1과 C가 각각 0.48mg/100g, 0.22mg/100g으로 나타났으며 지용성 비타민 E는 0.10mg/100g으로 확인되었으나 비타민 A, B12, D2, D3 및 K1은 검출되지 않았다. Referring to Table 4, the vitamins present in several pupae were found to be 0.48mg / 100g and 0.22mg / 100g, respectively, of water-soluble vitamins B 1 and C, and fat-soluble vitamin E was found to be 0.10mg / 100g, but vitamins A and B 12 , D 2 , D 3 and K 1 were not detected.
실시예 2-5. 지방산 함량분석Example 2-5. Fatty acid content analysis
지방산은 지방의 가수분해물로 탄화수소 사슬에 카르복실기를 가지는 화합물을 일컫는데, 탄화수소 사슬에 탄소-탄소 이중결합의 유무에 따라 포화지방산과 불포화지방산으로 나뉜다. 포화지방산은 체내에서 생합성 되며 동물성지방에 주로 함유되어 있는 물질로 알려져 있으나 식물성 팜유나 코코넛유에도 다량으로 존재한다(Verallo-Rowell et al., 2016). 포화지방산은 과잉 섭취시 심혈관계 질환을 유발하는 물질로도 알려져 있다(Mozaffarian et al., 2010).Fatty acid is a hydrolyzate of fat, and refers to a compound having a carboxyl group in the hydrocarbon chain. It is divided into saturated fatty acids and unsaturated fatty acids depending on the presence or absence of a carbon-carbon double bond in the hydrocarbon chain. Saturated fatty acids are biosynthesized in the body and are known to be mainly contained in animal fats, but are also present in large quantities in vegetable palm oil or coconut oil (Verallo-Rowell et al ., 2016). Saturated fatty acids are also known to cause cardiovascular disease when consumed excessively (Mozaffarian et al ., 2010).
서양종꿀벌의 수번데기에 존재하는 포화지방산과 불포화지방산의 함량은 식품공전의 지방산 시험법에 따라 실시하였다. 즉, 검체를 균질화하고 클로로포름과 디에틸에테르로 추출한 지방을 녹인 다음 질소로 농축하였다. 농축물에 트리플루오로보란메탄올과 톨루엔을 첨가하고 100℃ 오븐에서 가열 후 실온에서 냉각하였다. 이후 물과 헥산을 첨가하고 분리된 상층액을 취하여 시험용액으로 사용하였으며 불꽃이온화 검출기가 장착된 GC(Agilent 7890A)에 capillary column(Sigma-Aldrich SP-2560, St.Louis, MO, USA)과 검출온도 285℃, 흐름속도 0.75mL/min, 주입량 1μL, 오븐온도 100℃(4min), 208℃(3℃/min), 244℃(15min)의 분석조건으로 정량분석을 실시하였다. The content of saturated fatty acids and unsaturated fatty acids present in several pupae of Western honeybees was carried out according to the fatty acid test method of food industry. That is, the sample was homogenized, the fat extracted with chloroform and diethyl ether was dissolved, and then concentrated with nitrogen. Trifluoroboranmethanol and toluene were added to the concentrate, heated in an oven at 100 ° C., and then cooled at room temperature. Subsequently, water and hexane were added and the separated supernatant was used as a test solution and detected with a capillary column (Sigma-Aldrich SP-2560, St.Louis, MO, USA) in a GC (Agilent 7890A) equipped with a flame ionization detector. Quantitative analysis was performed under the analysis conditions of temperature 285 ° C, flow rate 0.75 mL / min,
표 5의 결과를 참고하면, 수번데기에 존재하는 포화지방산은 3.14g/100g으로, 주요 단백질원으로 알려진 돼지고기(삼겹살 기준, 15.47%) 및 소고기(4.67%)의 포화지방산 함량보다 적을 뿐만 아니라 한국인의 밥상에 자주 오르는 생성인 고등어(3.96%)의 함유량보다 낮은 수준이며(농촌진흥청, 2011), 불포화지방산이 2.31g/100g으로 2%정도 함유되어 있어 식품원료로 충분히 이용할 수 있을 것으로 파악된다.Referring to the results in Table 5, the saturated fatty acid present in several pupae is 3.14g / 100g, which is not only less than the saturated fatty acid content of pork (based on pork belly, 15.47%) and beef (4.67%), which are known as major protein sources. It is lower than the content of mackerel (3.96%), a product that frequently rises on Korean meals (Rural Development Administration, 2011), and contains 2% of unsaturated fatty acid at 2.31g / 100g, so it can be used as a food ingredient. .
<실시예 3. 식용가능한 서양종꿀벌의 수번데기 분말 제조 및 관능평가> <Example 3. Preparation of edible western honeybee powder and sensory evaluation>
서양종꿀벌의 수번데기를 세척한 뒤 물기를 제거(탈수)한 후, 121℃에서 30분간 멸균하여 익힌 것을 수분함량이 8~15%가 되게 동결건조하고 분말로 제조하였다. 이 분말에 대해 맛과 향, 기호도 등을 누에의 번데기 분말과 비교하여 50명에게 평가하여 표 6에 5점 타점법(5점:매우좋음 - 1점:매우나쁨)으로 나타내었다. 그 결과 누에 번데기 분말과 유사한 기호도와 맛, 향을 갖는 것으로 확인되었다.After washing several pupae of the Western honey bee and removing (dehydrating) it, it was sterilized at 121 ° C for 30 minutes and lyophilized to a water content of 8-15%, and then prepared as a powder. About this powder, the taste, aroma, preference, etc. were compared to silkworm pupae powder and evaluated to 50 persons, and the result was shown in Table 6 by a 5-point scoring method (5 points: very good-1 point: very bad). As a result, it was confirmed that the silkworm pupa powder had similar taste, taste, and aroma.
<실시예 4. 서양종꿀벌 수번데기의 항산화 효능 확인> <Example 4. Confirmation of anti-oxidative efficacy of Western honeybee pupae>
항산화 활성은 DPPH법을 사용하여 비타민 C를 양성대조군으로 사용하여 측정하였다. 서양종꿀벌의 수번데기(수벌 번데기), 수벌 유충, 일벌 유충, 일벌 번데기를 세척한 뒤 물기를 제거(탈수)한 후, 수분함량이 8~15%가 되게 동결건조하고 분말로 제조하였다. 분말화된 수번데기(수벌 번데기), 수벌 유충, 일벌 유충, 일벌 번데기를 각각 1kg 기준 10ℓ의 에탄올을 10시간 동안 가한 뒤, 8000rpm에서 10분간 원심분리하여 추출액만을 취하고, 이 추출액을 감압농축 및 건조하여 서양종꿀벌 수번데기(수벌 번데기), 수벌 유충, 일벌 유충, 일벌 번데기의 에탄올 추출물을 얻었다. Antioxidant activity was measured using vitamin C as a positive control using the DPPH method. After washing the pupae (puppet pupae), the pupae pupae, the bee pupae, and the pupae pupae of the Western honey bee, after removing the water (dehydration), it was freeze-dried to a water content of 8-15% and prepared as a powder. Powdered pupae (pupa pupae), pupa larvae, pupa larvae, pupa pupae were added 10 l of ethanol for 10 hours each for 10 hours, centrifuged at 8000 rpm for 10 minutes to take only the extract, concentrated and dried under reduced pressure. To obtain ethanol extracts of Western honey bee pupae (puppet pupae), bee larvae, bee larvae, and bee pupae.
DPPH(1,1-Diphenyl-2-picrylhydrazyl) 라디칼 소거능은 Cho 등(2011)의 방법을 이용하여 측정하였다. 각 에탄올 추출물을 농도별로 에탄올로 용해시킨 시료 0.1 ㎖와 0.4 mM DPPH 용액 0.1 ㎖를 혼합한 뒤, 30분 동안 암소에서 반응시켰다. Microplate reader(Molecular Devices, Sunnyvale, CA, USA)의 515 nm에서 흡광도 값을 측정하였으며, DPPH 라디칼 소거능은 다음의 식을 이용하여 계산하였다.DPPH (1,1-Diphenyl-2-picrylhydrazyl) radical scavenging ability was measured using the method of Cho et al. (2011). Each ethanol extract was mixed with 0.1 ml of a sample dissolved in ethanol for each concentration and 0.1 ml of a 0.4 mM DPPH solution, and then reacted in the dark for 30 minutes. The absorbance value was measured at 515 nm of a microplate reader (Molecular Devices, Sunnyvale, CA, USA), and the DPPH radical scavenging ability was calculated using the following equation.
DPPH 라디칼 소거능(%) = [(Ac-As) / Ac] × 100DPPH radical scavenging capacity (%) = [(Ac-As) / Ac] × 100
Ac: 공시료 흡광도, As: 시료군 흡광도Ac: sample absorbance, As: sample group absorbance
그 결과 도 1의 결과와 같이 서양종꿀벌의 수번데기(수벌번데기) 추출물이 수벌의 유충이나 일벌 유충, 일벌 번데기 각각의 추출물에 비해 DPPH를 억제하는 항산화 활성이 현저하게 우수함을 확인할 수 있다. As a result, it can be confirmed that the antioxidant activity of inhibiting DPPH is significantly higher than that of the extracts of the pupae pupae (pupa pupae) of the honey bee, bee larvae, bee larvae, and bee pupae, as shown in FIG. 1.
<실시예 5. 세포 독성 확인><Example 5. Confirmation of cytotoxicity>
실시예 4에서 제조한 서양종꿀벌의 수번데기 추출물을 이용하여 세포 독성을 확인하였다. 이를 위해, 렛(Rat) 적혈구 세포에 대한 세포 독성을 확인하였는데, 적혈구 세포(Rat erythrocytes)를 인산-염화나트륨 완충액(PBS, pH 7.4)으로 세 번 세척한 후, 인산-염화나트륨 완충액(PBS, pH 7.4)으로 희석하여, 5.0%의 적혈구 세포 용액을 제조하였다. 그런 후, 5.0% 적혈구 세포 용액을 96-웰 플레이트에 100㎕씩 분주한 후에 각 추출물을 단계적으로 희석한 용액을 섞어주었다. 이어서 모든 튜브에 총액의 양이 200㎕가 되도록 인산-염화나트륨 완충액(PBS, pH 7.4)을 넣어주고, 37℃ 배양기에서 30분 동안 배양하였다. 상기 배양이 끝난 후, 원심 분리기에서 1000rpm의 회전 속도로 10분간 원심 분리하여 상등액을 96웰 플레이트로 옮겼다. 적혈구 세포에 대한 파괴능은 ELISA 판독기로 550nm의 파장 하에서의 흡광도를 측정하여 파괴능을 분석하였다. 그리고 대조군으로 0.1%(w/v) 트리톤 X-100으로 처리하였을 경우의 값을 100% 파괴능으로 계산하였으며, 적혈구 세포만을 넣은 경우를 0 % 파괴능으로 계산하였다. 실험결과 본 발명의 추출물은 1~1000㎍/㎖의 농도에서 적혈구 세포의 파괴능이 없음을 확인하였다(그래프 미기재). Cytotoxicity was confirmed by using the pupa extract of the Western honey bee prepared in Example 4. To this end, cytotoxicity to rat red blood cells was confirmed. After washing red blood cells (Rat erythrocytes) three times with phosphate-sodium chloride buffer (PBS, pH 7.4), phosphate-sodium chloride buffer (PBS, pH 7.4) ) To prepare a 5.0% red blood cell solution. Then, 100% of the 5.0% red blood cell solution was dispensed into a 96-well plate, followed by mixing the diluted solution of each extract step by step. Subsequently, phosphate-sodium chloride buffer (PBS, pH 7.4) was added so that the total amount of the total solution was 200 μl in all tubes, and the cells were incubated for 30 minutes in a 37 ° C. incubator. After the incubation, the supernatant was transferred to a 96-well plate by centrifugation for 10 minutes at a rotation speed of 1000 rpm in a centrifuge. The destruction ability for red blood cells was analyzed by measuring the absorbance under a wavelength of 550 nm with an ELISA reader. In addition, the value when treated with 0.1% (w / v) Triton X-100 as a control was calculated as 100% destructive ability, and the case where only red blood cells were added was calculated as 0% destructive ability. As a result of the experiment, it was confirmed that the extract of the present invention had no destruction ability of red blood cells at a concentration of 1 to 1000 µg / ml (not shown in the graph).
<제제예 1. 약학적 제제 - 정제의 제조><Formulation Example 1. Pharmaceutical preparation-Preparation of tablet>
본 발명의 서양종꿀벌 수번데기의 에탄올 추출물 200g을 락토오스 175.9g, 감자전분 180g 및 콜로이드성 규산 32g과 혼합하였다. 이 혼합물에 10% 젤라틴 용액을 첨가시킨 후, 분쇄해서 14 메쉬체를 통과시켰다. 이것을 건조시키고 여기에 감자전분 160g, 활석 50g 및 스테아린산 마그네슘 5g을 첨가해서 얻은 혼합물을 정제로 만들었다. 200 g of ethanol extract of Western honeybee pupae of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. After adding 10% gelatin solution to this mixture, it was ground and passed through a 14 mesh sieve. This mixture was dried, and a mixture obtained by adding potato starch 160g, talc 50g, and magnesium stearate 5g was purified.
<제제예 2. 식품 제조><Formulation Example 2. Food manufacturing>
제제예 2-1. 조리용 양념의 제조Formulation Example 2-1. Cooking seasoning
본 발명의 멸균된 서양종꿀벌 수번데기의 동결건조 분말을 조리용 양념에 1 중량%로 첨가하여 건강 증진용 조리용 양념을 제조하였다.The freeze-dried powder of sterilized Western honey bee pupae of the present invention was added to the cooking seasoning in 1% by weight to prepare a cooking seasoning for promoting health.
제제예 2-2. 밀가루 식품의 제조Formulation Example 2-2. Production of flour food
본 발명의 멸균된 서양종꿀벌 수번데기의 동결건조 분말을 밀가루에 0.1 중량%로 첨가하고, 이 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하여 건강 증진용 식품을 제조하였다.The lyophilized powder of sterilized Western honey bee pupae of the present invention was added to wheat flour at 0.1% by weight, and bread, cake, cookies, crackers and noodles were prepared using this mixture to prepare health foods.
제제예 2-3. 스프 및 육즙(gravies)의 제조Formulation Example 2-3. Preparation of soup and gravy
본 발명의 멸균된 서양종꿀벌 수번데기의 동결건조 분말을 스프 및 육즙에 0.1 중량%로 첨가하여 건강 증진용 수프 및 육즙을 제조하였다.The lyophilized powder of sterilized Western honey bee pupae of the present invention was added to soups and gravy at 0.1% by weight to prepare health soup and gravy.
제제예 2-4. 유제품(dairy products)의 제조Formulation Example 2-4. Manufacturing dairy products
본 발명의 서양종꿀벌 수번데기의 에탄올 추출물을 우유에 0.1 중량%로 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.Ethanol extract of Western honeybee pupae of the present invention was added to milk at 0.1% by weight, and various dairy products such as butter and ice cream were prepared using the milk.
제제예 2-5. 야채주스 제조Formulation Example 2-5. Vegetable juice production
본 발명의 서양종꿀벌 수번데기의 에탄올 추출물 0.5g을 토마토주스 또는 당근주스 1,000㎖에 가하여 건강 증진용 야채주스를 제조하였다.0.5 g of ethanol extract of Western honeybee pupae of the present invention was added to tomato juice or carrot juice to prepare a vegetable juice for health promotion.
제제예 2-6. 과일주스 제조Formulation Example 2-6. Fruit juice manufacturing
본 발명의 서양종꿀벌 수번데기의 에탄올 추출물 0.1g을 사과주스 또는 포도주스 1,000㎖에 가하여 건강 증진용 과일주스를 제조하였다.0.1 g of ethanol extract of Western honeybee pupae of the present invention was added to 1,000 ml of apple juice or grape juice to prepare a fruit juice for health promotion.
<제제예 3. 화장료 제형 제조><Formulation Example 3. Preparation of cosmetic formulation>
표 7의 성분을 혼합하여 에멀전 조성물을 제조하였고, 피부자극 실험을 통해 무자극을 유지하는 조성물임을 확인하였다. The composition of Table 7 was mixed to prepare an emulsion composition, and it was confirmed that the composition maintains irritation through a skin irritation experiment.
Claims (10)
상기 추출물은 서양종꿀벌 수번데기를 물, C1~C4 알코올 또는 이들의 혼합용액을 용매로 하여 추출하는 것을 특징으로 하는 항산화용 화장료 조성물.According to claim 1,
The extract is a cosmetic composition for antioxidant, characterized in that the Western honeybee pupae is extracted using water, C1 to C4 alcohol, or a mixed solution thereof.
상기 C1~C4 알코올은 메탄올, 에탄올, 프로판올, 이소프로판올, 부탄올 및 이소부탄올로 이루어진 군에서 선택되는 것을 특징으로 하는 항산화용 화장료 조성물.According to claim 2,
The C1 ~ C4 alcohol is a cosmetic composition for antioxidant, characterized in that is selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.
상기 추출물은 서양종꿀벌 수번데기를 물, C1~C4 알코올 또는 이들의 혼합용액을 용매로 하여 추출하는 것을 특징으로 하는 항산화용 조성물.The method of claim 4,
The extract is a composition for antioxidant, characterized in that the Western honeybee pupae is extracted using water, C1 to C4 alcohol, or a mixed solution thereof.
상기 C1~C4 알코올은 메탄올, 에탄올, 프로판올, 이소프로판올, 부탄올 및 이소부탄올로 이루어진 군에서 선택되는 것을 특징으로 하는 항산화용 조성물.The method of claim 5,
The C1 ~ C4 alcohol is an antioxidant composition, characterized in that selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.
상기 수번데기를 물, C1~C4 알코올 또는 이들의 혼합용액을 용매로 하여 추출하는 것을 특징으로 하는 항산화용 건강기능식품.The method of claim 7,
Antioxidant health functional food, characterized in that the pupae is extracted with water, C1 to C4 alcohol or a mixed solution thereof.
상기 C1~C4 알코올은 메탄올, 에탄올, 프로판올, 이소프로판올, 부탄올 및 이소부탄올로 이루어진 군에서 선택되는 것을 특징으로 하는 항산화용 건강기능식품.The method of claim 8,
The C1 ~ C4 alcohol is a health functional food for antioxidant, characterized in that selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.
상기 수번데기를 건조한 후 분말화하는 단계; 및,
분말화된 수번데기에 물, C1~C4 알코올 또는 이들의 혼합용액을 가하여 추출한 후 건더기를 제거하여 얻은 여액을 농축 및 건조하는 단계;
를 포함하는 것을 특징으로 하는 서양종꿀벌 수번데기 추출물을 함유하는 항산화용 조성물의 제조방법. Washing and dehydrating several pupae of Western honey bee ( Apis mellifera );
Drying the pupae and pulverizing it; And,
Condensing and drying the filtrate obtained by extracting by adding water, C1 ~ C4 alcohol or a mixed solution thereof to the powdered pupae;
Method for producing an antioxidant composition containing a Western honey bee pupa extract characterized in that it comprises a.
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