KR20230019559A - Natural aroma composition using citrus flower and manufacturing method thereof - Google Patents
Natural aroma composition using citrus flower and manufacturing method thereof Download PDFInfo
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- KR20230019559A KR20230019559A KR1020210101205A KR20210101205A KR20230019559A KR 20230019559 A KR20230019559 A KR 20230019559A KR 1020210101205 A KR1020210101205 A KR 1020210101205A KR 20210101205 A KR20210101205 A KR 20210101205A KR 20230019559 A KR20230019559 A KR 20230019559A
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- citrus
- natural food
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/40—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by drying or kilning; Subsequent reconstitution
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- A—HUMAN NECESSITIES
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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
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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Abstract
Description
본 발명은 꽃을 이용하여 수입에 의존하고 있는 천연 식품향료를 대체할 수 있으며 화학물질을 이용하지 않아 인체에 유해하지 않은 천연 식품향료 및 이의 제조방법에 관한 것이다.The present invention relates to a natural food flavor that can replace natural food flavor that depends on imports using flowers and is not harmful to the human body because it does not use chemicals and a method for manufacturing the same.
향료는 식품, 화장품, 음료, 계면활성제, 사료, 화학, 바이오의약, 제약산업 등에 있어서 매우 중요한 역할을 하고 있다. 이와 같은 향료 생산 방법은 크게 화학합성법과 생물학적인 방법으로 나뉘는데, 화학적 합성방법을 통한 향료성분의 생산은 다양한 형태의 화학구조를 가진 향의 대량 생산이 가능하지만, 원하지 않은 라세믹 혼합물(racemic mixture)로 만들어질 가능성이 있을 뿐만 아니라, 기질특이성(substrate specificity)이 낮다는 단점이 있다. BACKGROUND OF THE INVENTION Spices play a very important role in food, cosmetics, beverages, surfactants, feeds, chemicals, biopharmaceuticals, and pharmaceutical industries. The production method of such a fragrance is largely divided into a chemical synthesis method and a biological method. Production of a fragrance ingredient through a chemical synthesis method is capable of mass production of fragrance with various types of chemical structures, but it is possible to produce an undesirable racemic mixture. Not only is it possible to make it, but it also has the disadvantage of low substrate specificity.
아울러, 최근 소비자들의 소비 성향은 자연 친화적이고, 인체에 무해한 향취성분이 포함된 식품, 의약, 화학제품, 의약품제품을 선호하는 방향으로 변화하고 있고, 이에 따라 화학합성법으로 생산된 향료 혹은 향취성분보다는 생물학적 방법으로 생산한 향료성분에 대해 많은 관심이 높아지고 있다.In addition, consumers' consumption propensity is changing in the direction of preferring foods, medicines, chemical products, and pharmaceutical products that are environmentally friendly and contain fragrance ingredients that are harmless to the human body. There is a growing interest in fragrance ingredients produced by biological methods.
세계 향료시장은 건강에 대한 관심증가로 천연향료 소재 개발을 통해 건강기능성을 가미한 제품을 출시하고 있으나, 국내 향료업체는 합성향료 원료를 배합하는 수준에 머물러 있고, 천연향료 연구는 기능성 부분에만 초점을 맞추어 식품 개발을 위한 적용 및 대량 생산기술 개발이 전무한 실정이다.The global fragrance market is launching products with health functionalities added through the development of natural fragrance materials due to the growing interest in health. Accordingly, there is no application for food development and development of mass production technology.
현재 향료시장의 규모는 세계적으로 약 100억 달러이고, 국내 시장은 약 1억 3천만 달러에 이르지만, 국내의 경우에는 전체 시장의 80% 이상을 외국 업체들이 점유하고 있으며, 천연향료는 거의 100% 정도 수입에 의존하고 있다.Currently, the size of the spice market is about 10 billion dollars worldwide, and the domestic market is about 130 million dollars. It is somewhat dependent on income.
특히, 국내 다소비 향(시트러스 계열)이 주로 수입됨에도 불구하고 국내에서 수행된 기존연구는 국내 농산물 유래 허브, 두류, 약용, 한약재 식물위주로 연구되었을 뿐 시트러스 과일 열매 등의 국내 자생소재를 이용한 천연 향료에 대한 개발이 이루어지고 있지 않고 있다. In particular, despite the fact that domestically consumed fragrances (citrus series) are mainly imported, existing studies conducted in Korea have focused on herbs derived from domestic agricultural products, pulses, medicinal plants, and herbal medicinal plants. Natural fragrances using domestic materials such as citrus fruits is not being developed.
더욱이, 식품향료의 경우에는 일본에 대한 수입 의존률이 70% 정도로 매우 높으며, 일본은 우리나라와 식생활의 유사성과 향료 선호경향이 높아 국내 향료 분야에서 강점을 보이고 있다.Moreover, in the case of food spices, the dependence on imports from Japan is very high at about 70%, and Japan shows strength in the field of domestic spices because of its similarity in dietary life and preference for spices with Korea.
이에 따라, 외국으로부터 대량 수입되는 식품첨가물인 천연 향료를 대체하기 위해 향미소재 특성 규명 통한 소재 발굴, 천연향료 개발을 통한 국산화 노력이 시급하다.Accordingly, in order to replace natural spices, which are food additives that are imported in large quantities from abroad, it is urgent to develop materials through the identification of flavor material characteristics and localization efforts through the development of natural flavors.
본 발명의 목적은 수입에 의존하고 있는 천연 식품향료를 대체할 수 있으며 화학물질을 이용하지 않아 인체에 유해하지 않은 천연 식품향료를 제공하는데 있다.An object of the present invention is to provide natural food flavors that can replace natural food flavors dependent on imports and are not harmful to the human body because they do not use chemicals.
또한, 본 발명의 다른 목적은 상기 천연 식품향료를 제조하는 방법을 제공하는데 있다.In addition, another object of the present invention is to provide a method for producing the natural food flavoring.
또한, 본 발명의 또 다른 목적은 상기 천연 식품향료를 포함하는 식품 조성물을 제공하는데 있다.In addition, another object of the present invention is to provide a food composition containing the natural food flavoring.
또한, 본 발명의 또 다른 목적은 천연 식품향료를 유효성분으로 함유하는 피부 미백용 건강기능식품을 제공하는데 있다.In addition, another object of the present invention is to provide a health functional food for skin whitening containing natural food flavorings as active ingredients.
또한, 본 발명의 또 다른 목적은 천연 식품향료를 유효성분으로 함유하는 피부 미백용 화장료 조성물을 제공하는데 있다.In addition, another object of the present invention is to provide a cosmetic composition for skin whitening containing a natural food flavoring agent as an active ingredient.
상기한 목적을 달성하기 위한 본 발명의 천연 식품향료를 제조하는 방법은 (A) 꽃을 동결건조시키는 단계; (B) 상기 동결건조된 꽃을 분쇄한 후 30 내지 60 ℃에서 용매로 1차 추출하여 에센셜 오일을 수득하는 단계; 및 (C) 상기 1차 추출된 에센셜 오일을 2차로 초임계 추출하여 천연 식품향료를 수득하는 단계;를 포함할 수 있다.Method for producing a natural food flavoring of the present invention for achieving the above object is (A) freeze-drying the flower; (B) obtaining essential oil by first extracting the freeze-dried flowers with a solvent at 30 to 60 ° C.; and (C) secondarily supercritical extraction of the firstly extracted essential oil to obtain a natural food flavor.
상기 (A)단계에서 동결건조는 -80 내지 -70 ℃에서 동결시켜 10 내지 20시간 동안 수행될 수 있다.Freeze-drying in step (A) may be performed for 10 to 20 hours by freezing at -80 to -70 ° C.
상기 (B)단계에서 1차 용매추출은 꽃과 용매를 혼합한 후 30 내지 60 ℃에서 40 내지 80분 동안 수행될 수 있다.In the step (B), the first solvent extraction may be performed at 30 to 60 ° C. for 40 to 80 minutes after mixing the flowers and the solvent.
상기 용매는 물, 탄소수 1 내지 4의 저급알코올, 에틸렌글리콜, 에틸에테르 또는 이들의 혼합용매일 수 있다.The solvent may be water, lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixture thereof.
상기 (C)단계에서 초임계 추출은 300 내지 400 bar의 압력 및 40 내지 70 ℃의 온도 하에서 1 내지 3시간 동안 수행될 수 있다.Supercritical extraction in step (C) may be performed for 1 to 3 hours under a pressure of 300 to 400 bar and a temperature of 40 to 70 °C.
상기 (C)단계에서 초임계 추출 시 용매는 에탄올일 수 있다.In the step (C), the solvent for supercritical extraction may be ethanol.
상기 꽃은 감귤류 꽃일 수 있다.The flower may be a citrus flower.
상기 천연 식품향료는 향기성분으로 D-리모넨(D-Limonene), γ-테르피넨(γ-Terpinene), β-오시멘(β-Ocimene), p-사이멘(p-cymene), β-엘레멘(β-elemene), 카요필렌(Caryophyllene), α-휴물렌(α-Humulene), 제르마크렌 D(Germacrene D), α-파네신(α-Farnesene) 및 벤질 니트릴(Benzyl nitrile)을 포함할 수 있다.The natural food flavors are fragrance components, which include D-Limonene, γ-Terpinene, β-Ocimene, p-cymene, and β-Ele Contains β-elemene, Caryophyllene, α-Humulene, Germacrene D, α-Farnesene and Benzyl nitrile can do.
상기 천연 식품향료는 향기성분으로 100 내지 300 ㎍/ml의 D-리모넨(D-Limonene), 700 내지 900 ㎍/ml의 γ-테르피넨(γ-Terpinene), 100 내지 300 ㎍/ml의 β-오시멘(β-Ocimene), 1000 내지 1500 ㎍/ml의 p-사이멘(p-cymene), 1200 내지 1600 ㎍/ml의 β-엘레멘(β-elemene), 1500 내지 2000 ㎍/ml의 카요필렌(Caryophyllene), 700 내지 900 ㎍/ml의 α-휴물렌(α-Humulene), 200 내지 400 ㎍/ml의 제르마크렌 D(Germacrene D), 600 내지 900 ㎍/ml의 α-파네신(α-Farnesene) 및 200 내지 400 ㎍/ml의 벤질 니트릴(Benzyl nitrile)을 포함할 수 있다.The natural food flavoring is a fragrance component, 100 to 300 μg / ml of D-Limonene (D-Limonene), 700 to 900 μg / ml of γ-terpinene (γ-Terpinene), 100 to 300 μg / ml of β- β-Ocimene, 1000 to 1500 μg/ml p-cymene, 1200 to 1600 μg/ml β-elemene, 1500 to 2000 μg/ml Cayo Caryophyllene, 700 to 900 μg/ml of α-Humulene, 200 to 400 μg/ml of Germacrene D, 600 to 900 μg/ml of α-farnesin ( α-Farnesene) and 200 to 400 μg/ml of benzyl nitrile.
상기 감귤류는 감귤(Citrus unshiu), 레몬(Citrus limon), 오렌지(Citrus sinensis), 유자(Citrus junos), 자몽(Citrus paradisi) 또는 한라봉(Citrus hybrid Shiranuhi)일 수 있다.The citrus may be citrus (Citrus unshiu), lemon (Citrus limon), orange (Citrus sinensis), citron (Citrus junos), grapefruit (Citrus paradisi), or hallabong (Citrus hybrid Shiranuhi).
또한, 상기한 다른 목적을 달성하기 위한 본 발명의 천연 식품향료는 꽃을 용매로 1차 추출하여 수득된 에센셜 오일을 초임계로 2차 추출한 것일 수 있다.In addition, the natural food flavoring of the present invention for achieving the above other object may be obtained by first extracting flowers with a solvent and secondarily extracting essential oil using a supercritical method.
또한, 상기한 또 다른 목적을 달성하기 위한 본 발명의 식품 조성물은 상기 천연 식품향료를 포함할 수 있다.In addition, the food composition of the present invention for achieving another object described above may include the natural food flavoring agent.
또한, 상기한 또 다른 목적을 달성하기 위한 본 발명의 건강기능식품은 상기 천연 식품향료를 유효성분으로 함유하는 피부 미백용 건강기능식품일 수 있다.In addition, the health functional food of the present invention to achieve the above another object may be a health functional food for skin whitening containing the natural food flavoring as an active ingredient.
또한, 상기한 또 다른 목적을 달성하기 위한 본 발명의 화장료 조성물은 상기 천연 식품향료를 유효성분으로 함유하는 피부 미백용 화장료 조성물일 수 있다.In addition, the cosmetic composition of the present invention for achieving the above another object may be a cosmetic composition for skin whitening containing the natural food flavoring agent as an active ingredient.
본 발명의 꽃을 이용한 천연 식품향료는 다량의 향기성분이 장시간 유지되어 소비자의 기호도를 만족시킬 수 있으므로 현재 수입에 의존하고 있는 천연 식품향료를 대체할 수 있으며 화학물질을 이용하지 않아 인체에 유해하지 않다.Natural food flavorings using flowers of the present invention can replace natural food flavorings currently dependent on imports because a large amount of fragrance components are maintained for a long time and can satisfy consumers' preference, and are not harmful to the human body because they do not use chemicals. not.
도 1A는 본 발명의 실시예 1에 따라 제조된 천연 식품향료가 처리된 제브라피쉬를 멜라닌 생성 억제를 나타낸 사진이며, 도 1B는 상기 도 1A의 제브라피쉬에 생성된 멜라닌을 정량화한 그래프이다.Figure 1A is a photograph showing the inhibition of melanin production in zebrafish treated with a natural food flavor prepared according to Example 1 of the present invention, Figure 1B is a graph quantifying melanin produced in the zebrafish of Figure 1A.
본 발명은 꽃을 이용하여 수입에 의존하고 있는 천연 식품향료를 대체할 수 있으며 화학물질을 이용하지 않아 인체에 유해하지 않은 천연 식품향료 및 이의 제조방법에 관한 것이다.The present invention relates to a natural food flavor that can replace natural food flavor that depends on imports using flowers and is not harmful to the human body because it does not use chemicals and a method for manufacturing the same.
이하, 본 발명을 상세하게 설명한다. Hereinafter, the present invention will be described in detail.
본 발명의 천연 식품향료를 제조하는 방법은 (A) 꽃을 동결건조시키는 단계; (B) 상기 동결건조된 꽃을 분쇄한 후 30 내지 60 ℃에서 용매로 1차 추출하여 에센셜 오일을 수득하는 단계; 및 (C) 상기 1차 추출된 에센셜 오일을 2차로 초임계 추출하여 천연 식품향료를 수득하는 단계;를 포함할 수 있다.The method for preparing a natural food flavoring of the present invention includes (A) freeze-drying a flower; (B) obtaining essential oil by first extracting the freeze-dried flowers with a solvent at 30 to 60 ° C.; and (C) secondarily supercritical extraction of the firstly extracted essential oil to obtain a natural food flavor.
먼저, 상기 (A)단계에서는 꽃을 -80 내지 -70 ℃에서 동결시켜 10 내지 20시간, 바람직하게는 12 내지 15시간 동안 동결건조시킨다.First, in the step (A), the flowers are frozen at -80 to -70 ° C and freeze-dried for 10 to 20 hours, preferably 12 to 15 hours.
동결건조는 추출 시 향기성분이 다량 추출될 수 있도록 하는 것으로서, 동결건조를 수행하지 않거나 열풍건조, 자연건조, 냉풍건조 등의 다른 건조방법을 수행하는 하는 경우에는 원하는 향기성분을 얻을 수 없을 뿐만 아니라 향기성분이 다량으로 추출되지 않는다.Freeze-drying allows a large amount of fragrance components to be extracted during extraction. If freeze-drying is not performed or other drying methods such as hot air drying, natural drying, and cold air drying are performed, desired fragrance components cannot be obtained. Fragrance components are not extracted in large quantities.
동결건조를 수행하는 시간이 상기 하한치 미만인 경우에는 원하는 향기성분을 얻을 수 없으며, 상기 상한치 초과인 경우에는 향기성분이 다량으로 추출되지 않는다.If the freeze-drying time is less than the lower limit, the desired fragrance component cannot be obtained, and if it exceeds the upper limit, the fragrance component is not extracted in large quantities.
본 발명에서 사용되는 꽃은 감귤류 꽃으로서, 상기 감귤류 꽃은 감귤(Citrus unshiu), 레몬(Citrus limon), 오렌지(Citrus sinensis), 유자(Citrus junos), 자몽(Citrus paradisi) 또는 한라봉(Citrus hybrid Shiranuhi)의 꽃을 들 수 있다. The flower used in the present invention is a citrus flower, and the citrus flower includes citrus (Citrus unshiu), lemon (Citrus limon), orange (Citrus sinensis), citron (Citrus junos), grapefruit (Citrus paradisi) or hallabong (Citrus hybrid Shiranuhi) ) flowers can be picked up.
다음으로, 상기 (B)단계에서는 상기 동결건조된 꽃을 분쇄한 후 용매로 1차 추출하여 에센셜 오일을 수득한다.Next, in the step (B), the lyophilized flowers are ground and extracted first with a solvent to obtain essential oil.
상기 1차 추출은 꽃 분쇄물을 추출용매로 추출한 추출물에서 오일층과 물층을 분리한 후 상기 오일층만 수득하여 다음 공정에 이용한다. In the primary extraction, the oil layer and the water layer are separated from the extract obtained by extracting the pulverized flower as an extraction solvent, and then only the oil layer is obtained and used in the next process.
구체적으로, 상기 꽃 분쇄물과 추출용매를 1 : 5 내지 25 중량비, 바람직하게는 1 : 8 내지 15의 중량비로 혼합하여 30 내지 60 ℃, 바람직하게는 40 내지 50 ℃에서 40 내지 80분, 바람직하게는 50 내지 60분 동안 추출하여 추출물을 제조한 다음 오일층과 물층을 분리하고 상기 오일층만 수득하여 다음 공정에 이용한다. Specifically, the pulverized flowers and the extraction solvent are mixed in a weight ratio of 1: 5 to 25, preferably 1: 8 to 15, and then heated at 30 to 60 ° C., preferably 40 to 50 ° C. for 40 to 80 minutes, preferably Preferably, an extract is prepared by extracting for 50 to 60 minutes, and then the oil layer and the water layer are separated, and only the oil layer is obtained and used in the next process.
상기 꽃 분쇄물과 추출용매의 중량비가 상기 범위를 벗어나는 경우에는 향기성분이 적은 양으로 추출될 수 있다.When the weight ratio of the pulverized flower and the extraction solvent is out of the above range, the fragrance component may be extracted in a small amount.
추출온도 및 추출시간이 상기 하한치 미만인 경우에는 오일층이 적게 수득되고, 오일층 내의 향기성분도 적은 양으로 수득될 수 있다.When the extraction temperature and extraction time are less than the above lower limit, a small amount of oil layer is obtained, and a small amount of fragrance components in the oil layer can be obtained.
상기 각 추출물을 추출하는 추출용매로는 물, 탄소수 1 내지 4의 저급알코올, 에틸렌글리콜, 에틸에테르 또는 이들의 혼합용매를 들 수 있으며; 상기 저급알코올로는 20 내지 80%의 메탄올, 에탄올, 부탄올 또는 프로판올을 들 수 있다.The extraction solvent for extracting each extract may include water, lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixture thereof; Examples of the lower alcohol include 20 to 80% methanol, ethanol, butanol or propanol.
상기 추출용매로는 특별히 한정하는 것은 아니지만 물로 추출 시 다량의 향기성분을 함유한 오일층을 수득할 수 있다.The extraction solvent is not particularly limited, but when extracted with water, an oil layer containing a large amount of fragrance components can be obtained.
다음으로, 상기 (C)단계에서는 상기 1차 추출된 에센셜 오일을 2차로 초임계 추출하여 천연 식품향료를 수득한다.Next, in the step (C), the firstly extracted essential oil is secondarily supercritically extracted to obtain a natural food flavor.
상기 1차 추출시 수득한 에센셜 오일을 다시 초임계로 추출함으로써, 향기성분이 다량 추출될 뿐만 아니라 추출된 향기성분이 오랜 시간 유지될 수 있는 천연 식품향료를 수득할 수 있다.By supercritical extraction of the essential oil obtained in the first extraction, it is possible to obtain natural food flavors in which a large amount of flavor components can be extracted and the extracted flavor components can be maintained for a long time.
상기 초임계 추출법은 300 내지 400 bar, 바람직하게는 350 내지 400 bar의 압력 및 40 내지 70 ℃, 바람직하게는 50 내지 60 ℃ 온도의 초임계 상태에서 1 내지 3시간, 바람직하게는 1 내지 2시간 동안 수행하여 상기 에센셜 오일을 추출한다. 이때 용매로는 특별히 한정되지 않지만, 높은 추출 효율, 우수한 향기성분의 함량 및 지속성을 위해서 에탄올을 사용하여 임계상태(에탄올의 임계온도: 240.9 ℃, 임계압력: 61.4 bar)에서 추출하는 것이 바람직하다. The supercritical extraction method is carried out in a supercritical state at a pressure of 300 to 400 bar, preferably 350 to 400 bar and a temperature of 40 to 70 ° C., preferably 50 to 60 ° C. for 1 to 3 hours, preferably 1 to 2 hours. while extracting the essential oil. At this time, the solvent is not particularly limited, but it is preferable to extract in a critical state (critical temperature of ethanol: 240.9 ° C., critical pressure: 61.4 bar) using ethanol for high extraction efficiency, excellent content and persistence of fragrance components.
상기 초임계 추출법은 수증기 증류 추출법 등의 다른 추출법으로 추출하는 경우에 비하여 추출 효율이 높고 더욱 우수한 향기성분의 함량 및 지속성을 가질 수 있다. 특히, 수증기 증류 추출법을 사용하는 경우에는 향취가 사라지고 열에 의해 변성된 좋지 못한 향취가 발생된다.The supercritical extraction method may have higher extraction efficiency and better content and durability of fragrance components compared to the case of extraction by other extraction methods such as steam distillation extraction method. In particular, in the case of using the steam distillation extraction method, the odor disappears and a bad odor denatured by heat is generated.
초임계 추출 시 압력이 상기 하한치 미만인 경우에는 향기성분이 다량 추출되지 못할 수 있으며, 상기 상한치 초과인 경우에는 향취가 사라질 수 있다. 또한, 상기 초임계 추출 시 온도 및 시간이 상기 하한치 미만인 경우에는 추출효율이 낮을 수 있으며, 상기 상한치 초과인 경우에는 좋지 못한 향취가 발생할 수 있다.When the pressure during supercritical extraction is less than the lower limit, a large amount of the fragrance component may not be extracted, and when the pressure exceeds the upper limit, the fragrance may disappear. In addition, when the temperature and time of the supercritical extraction are less than the lower limit value, the extraction efficiency may be low, and when the temperature and time are greater than the upper limit value, a bad odor may occur.
상기와 같이 제조된 본 발명의 감귤류 꽃을 이용한 천연 식품향료는 향기성분으로 100 내지 300 ㎍/ml의 D-리모넨(D-Limonene), 700 내지 900 ㎍/ml의 γ-테르피넨(γ-Terpinene), 100 내지 300 ㎍/ml의 β-오시멘(β-Ocimene), 1000 내지 1500 ㎍/ml의 p-사이멘(p-cymene), 1200 내지 1600 ㎍/ml의 β-엘레멘(β-elemene), 1500 내지 2000 ㎍/ml의 카요필렌(Caryophyllene), 700 내지 900 ㎍/ml의 α-휴물렌(α-Humulene), 200 내지 400 ㎍/ml의 제르마크렌 D(Germacrene D), 600 내지 900 ㎍/ml의 α-파네신(α-Farnesene) 및 200 내지 400 ㎍/ml의 벤질 니트릴(Benzyl nitrile)을 포함할 수 있다.The natural food flavoring using the citrus flower of the present invention prepared as described above contains 100 to 300 μg/ml of D-Limonene and 700 to 900 μg/ml of γ-terpinene as fragrance components. ), 100 to 300 μg/ml of β-Ocimene, 1000 to 1500 μg/ml of p-cymene, and 1200 to 1600 μg/ml of β-element (β- elemene), 1500 to 2000 μg/ml of Caryophyllene, 700 to 900 μg/ml of α-Humulene, 200 to 400 μg/ml of Germacrene D, 600 to 900 μg/ml of α-farnesene and 200 to 400 μg/ml of benzyl nitrile.
또한, 본 발명은 상기 천연 식품향료를 함유하는 식품 조성물을 제공할 수 있다.In addition, the present invention can provide a food composition containing the natural food flavor.
상기 식품조성물로는 천연 식품향료를 사용할 수 있는 제품이라면 특별히 한정되지 않지만, 바람직하게는 청량음료, 발효유, 빵, 국수, 과자, 빙과류, 소시지, 치즈, 식용가공식품 또는 수산물가공식품을 들 수 있다.The food composition is not particularly limited as long as it can use natural food flavoring, but preferably includes soft drinks, fermented milk, bread, noodles, confectionery, frozen confectionery, sausages, cheese, edible processed foods, or processed seafood products. .
또한, 본 발명의 천연 식품향료는 멜라민의 생성을 억제시킬 수 있으므로 피부 미백용 건강기능식품 또는 피부 미백용 화장료 조성물로 이용될 수 있다. In addition, since the natural food flavor of the present invention can inhibit the production of melamine, it can be used as a health functional food for skin whitening or a cosmetic composition for skin whitening.
상기 ‘식품 조성물’은 유효성분으로 천연 식품향료 이외에, 식품 제조에 통상적으로 사용되는 식품의 기준 및 규격(‘식품공전’)에 기재된 식품으로 사용가능한 식품 원료, 식품첨가물 공전에 기재된 식품첨가물을 포함한다.The 'food composition' includes, in addition to natural food flavors as active ingredients, food raw materials that can be used as food described in the standards and specifications of food commonly used in food manufacturing ('Food Code'), and food additives described in the Food Additive Code. do.
특별히 한정할 필요는 없으나 예를 들어 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함한다. 상기 탄수화물은 단당류, 예를 들어, 포도당, 과당 등; 이당류, 예를 들어 말토스, 설탕, 유당 등; 올리고당 또는 폴리사카라이드, 예를 들어 덱스트린, 물엿, 사이클로덱스트린 등; 당알코올, 예를 들어 자일리톨, 소르비톨, 에리트리톨 등을 사용할 수 있다. 상기 향미제는 천연 향미제[타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다.It does not need to be particularly limited, but includes, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. The carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sugar, lactose and the like; oligosaccharides or polysaccharides such as dextrin, starch syrup, cyclodextrin and the like; Sugar alcohols such as xylitol, sorbitol, erythritol and the like can be used. As the flavoring agent, natural flavoring agents (thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be used.
상기 천연 식품향료를 유효성분으로 식품 조성물을 제조하는 경우 천연 식품향료는 다량의 향기성분이 장시간 지속될 수 있는 함량이면 특별히 한정할 필요는 없으나, 예를 들어 0.1 내지 99 중량%, 0.5 내지 95 중량%, 1 내지 90 중량%, 1 내지 80 중량%, 1 내지 70 중량%, 1 내지 60 중량%, 1 내지 50 중량%로 포함될 수 있다.When preparing a food composition using the natural food flavor as an active ingredient, the natural food flavor does not need to be particularly limited as long as a large amount of the flavor component can last for a long time, but, for example, 0.1 to 99% by weight, 0.5 to 95% by weight , 1 to 90% by weight, 1 to 80% by weight, 1 to 70% by weight, 1 to 60% by weight, may be included in 1 to 50% by weight.
상기 식품 조성물에서 유효성분인 천연 식품향료는 섭취자의 상태, 체중, 질병의 유무나 정도 및 기간에 따라 다르지만, 통상의 기술자에 의해 적절하게 선택될 수 있다. 예들 들어 1일 투여량을 기준으로 1 내지 5,000 mg, 바람직하게는 5 내지 2,000 mg, 더욱 바람직하게는 10 내지 1,000 mg, 더더욱 바람직하게는 20 내지 800 mg, 가장 바람직하게는 50 내지 500 mg일 수 있고, 투여 횟수는 특별히 한정할 필요는 없으나 1일 3회 내지 1주일에 1회의 범위 내에서 통상의 기술자가 조절할 수 있다. Natural food flavorings, which are active ingredients in the food composition, vary depending on the condition, body weight, presence or absence, degree, and period of disease of the eater, but may be appropriately selected by a person skilled in the art. For example, it may be 1 to 5,000 mg, preferably 5 to 2,000 mg, more preferably 10 to 1,000 mg, still more preferably 20 to 800 mg, and most preferably 50 to 500 mg based on the daily dose. There is, and the number of administrations need not be particularly limited, but can be adjusted by a person skilled in the art within the range of three times a day to once a week.
상기 식품 조성물은 특별히 한정할 필요는 없으나 예를 들어 산제, 과립제, 정제, 캡슐제, 환제, 엑스제, 젤리 제형, 티백 제형 또는 음료 제형일 수 있다.The food composition does not need to be particularly limited, but may be, for example, a powder, granule, tablet, capsule, pill, extract, jelly formulation, tea bag formulation or beverage formulation.
또한 일반 식품에 향기성분을 부여하기 위하여 상기 천연 식품향료를 첨가할 수 있으며, 첨가가 가능한 식품은, 특별히 한정할 필요는 없으나 예를 들어 식품위생법 제7조에 따른 식품의 기준 및 규격(‘식품공전’)에 예시된 과자류, 빵 또는 떡류, 코코아가공품류 또는 초콜릿류, 식육 또는 알가공품, 어육가공품, 두부류 또는 묵류, 면류, 다류, 커피, 음료류, 특수용도식품, 장류, 조미식품, 드레싱류, 김치류, 젓갈류, 절임식품, 조림식품, 주류, 건포류, 기타 식품류 등에 첨가될 수 있다. 또한 축산물위생관리법 제4조에 따른 축산물의 가공기준 및 성분규격(‘축산물공전’)에 예시된 유가공품, 식육가공품 및 포장육, 알가공품에 첨가될 수 있다.In addition, the natural food flavoring agent can be added to general food to impart a fragrance component, and the food that can be added does not need to be particularly limited, but for example, the standards and specifications of food according to Article 7 of the Food Sanitation Act Confectionery, bread or rice cakes, processed cocoa products or chocolates, processed meat or egg products, processed fish meat products, tofu or jelly, noodles, teas, coffee, beverages, special purpose foods, soy sauces, seasoning foods, dressings, It can be added to kimchi, salted fish, pickled foods, stewed foods, alcoholic beverages, raisins, and other foods. In addition, it can be added to dairy products, processed meat products, packaged meat, and egg products exemplified in the processing standards and ingredient specifications of livestock products ('livestock product code') according to Article 4 of the Livestock Products Sanitation Control Act.
한편, 상기 천연 식품향료는 멜라민의 생성을 억제시킬 수 있으므로 단독으로 “피부 미백용 건강기능식품”으로 이용될 수 있다. On the other hand, since the natural food flavor can inhibit the production of melamine, it can be used alone as a “health functional food for skin whitening”.
상기 ‘건강기능식품’은 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 법적 기준에 따라 제조(가공을 포함)한 식품(건강기능식품에 관한 법률 제3조 제1호)을 말한다. 상기 ‘건강기능식품’은 국가마다 용어나 범위에 차이가 있을 수 있으나, 미국의 ‘식이 보충제(Dietary Supplement)’, 유럽의 ‘식품 보충제(Food Supplemnet)’, 일본의 ‘보건기능식품’ 또는 ‘특정보건용식품(Food for Special Health Use, FoSHU)’, 중국의 ‘보건식품’ 등에 해당할 수 있다.The above ‘functional health food’ refers to food manufactured (including processing) in accordance with legal standards using raw materials or ingredients that have functional properties useful for the human body (Article 3, Subparagraph 1 of the Health Functional Food Act). The term 'health functional food' may differ in terminology or scope from country to country, but 'Dietary Supplement' in the US, 'Food Supplement' in Europe, 'Health Functional Food' or 'Health Functional Food' in Japan. It may correspond to 'Food for Special Health Use (FoSHU)' or 'Health Food' in China.
상기 식품 조성물 또는 건강기능식품은 식품첨가물을 추가로 포함할 수 있으며, 식품첨가물로서의 적합여부는 다른 규정이 없는 한 ‘식품첨가물공전’의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 따른다.The food composition or health functional food may additionally contain food additives, and the suitability as a food additive is determined according to the standards and standards for the item in accordance with the general rules and general test methods of the 'Food Additive Code' unless otherwise specified. follow
또한, 상기 천연 식품향료는 멜라민의 생성을 억제시킬 수 있으므로 단독으로 “피부 미백용 화장료 조성물”로 이용될 수 있다. In addition, since the natural food flavor can inhibit the production of melamine, it can be used alone as a “cosmetic composition for skin whitening”.
본 발명의 피부 미백용 화장료 조성물은 일반적인 유화 제형 및 가용화 제형의 형태로 제조할 수 있다. 유화 제형의 화장품으로는 영양화장수, 크림, 에센스 등이 있으며, 가용화 제형의 화장품으로는 유연화장수가 있다.The cosmetic composition for skin whitening of the present invention can be prepared in the form of a general emulsified formulation and a solubilized formulation. Emulsified cosmetics include nutrition lotion, cream, essence, etc., and solubilized cosmetics include softening lotion.
적합한 화장품의 제형으로는 예를 들면 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀), 비이온형의 소낭 분산제의 형태, 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실 스틱의 형태로 제공될 수 있다. 또한, 포말(foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 제조될 수 있다.Suitable cosmetic formulations include, for example, solutions, gels, solid or pasty anhydrous products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionic vesicles. It may be provided in the form of a dispersant, cream, toner, lotion, powder, ointment, spray or conceal stick. In addition, it can be prepared in the form of a foam (foam) or the form of an aerosol composition further containing a compressed propellant.
또한, 상기 화장품은 본 발명의 천연 식품향료에 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다.In addition, the cosmetics include, in addition to the natural food flavoring agent of the present invention, a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, Water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or commonly used in cosmetics It may contain adjuvants commonly used in the cosmetic field, such as any other ingredients that are
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시하나, 하기 실시예는 본 발명을 예시하는 것일 뿐 본 발명의 범주 및 기술사상 범위 내에서 다양한 변경 및 수정이 가능함은 당업자에게 있어서 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속하는 것도 당연한 것이다.Hereinafter, preferred embodiments are presented to aid understanding of the present invention, but the following examples are merely illustrative of the present invention, and various changes and modifications are possible within the scope and spirit of the present invention. It is obvious to those skilled in the art, It goes without saying that these variations and modifications fall within the scope of the appended claims.
실시예 1.Example 1.
감귤의 꽃을 -78 ℃에서 동결시켜 12시간 동안 동결건조시킨 후 상기 동결건조된 감귤류 꽃을 분쇄하여 감귤류 꽃 분쇄물과 물을 1 : 10의 중량비로 혼합한 다음 45 ℃에서 60분 동안 추출하여 추출물을 수득하였다. 상기 추출물에서 오일층과 물층을 분리하여 오일층만 수득한 후 상기 오일층을 초임계 추출기에 넣은 다음 에탄올을 주입하여 50 ℃ 및 350 bar의 초임계 상태 하에서 2시간 동안 추출하여 천연 식품향료를 수득하였다.Citrus flowers were frozen at -78 ° C and freeze-dried for 12 hours, and then the lyophilized citrus flowers were pulverized, mixed with crushed citrus flowers and water in a weight ratio of 1: 10, and then extracted at 45 ° C for 60 minutes. An extract was obtained. After separating the oil layer and the water layer from the extract to obtain only the oil layer, the oil layer was put into a supercritical extractor, and then ethanol was injected to extract for 2 hours under a supercritical condition of 50 ° C. and 350 bar to obtain natural food flavor. did
비교예 1. 초임계 추출 생략Comparative Example 1. Supercritical extraction omitted
감귤의 꽃을 -78 ℃에서 동결시켜 12시간 동안 동결건조시킨 후 상기 동결건조된 감귤류 꽃을 분쇄하여 감귤류 꽃 분쇄물과 물을 1 : 10의 중량비로 혼합한 다음 45 ℃에서 60분 동안 추출하여 추출물을 수득하였다. 상기 추출물에서 오일층과 물층을 분리하여 오일층만 수득하여 천연 식품향료로 이용하였다.Citrus flowers were frozen at -78 ° C and freeze-dried for 12 hours, and then the lyophilized citrus flowers were pulverized, mixed with crushed citrus flowers and water in a weight ratio of 1: 10, and then extracted at 45 ° C for 60 minutes. An extract was obtained. The oil layer and the water layer were separated from the extract, and only the oil layer was obtained and used as a natural food flavoring.
비교예 2. 용매 추출 생략Comparative Example 2. Solvent extraction omitted
감귤의 꽃을 -78 ℃에서 동결시켜 12시간 동안 동결건조시킨 후 상기 동결건조된 감귤류 꽃을 분쇄하여 감귤류 꽃 분쇄물을 초임계 추출기에 넣은 다음 에탄올을 주입하여 50 ℃ 및 350 bar의 초임계 상태 하에서 2시간 동안 추출하여 천연 식품향료를 수득하였다.After freezing the citrus flowers at -78 ° C and freeze-drying them for 12 hours, the freeze-dried citrus flowers were crushed and the crushed citrus flowers were put into a supercritical extractor, and then ethanol was injected into a supercritical state at 50 ° C and 350 bar. Natural food flavor was obtained by extraction for 2 hours under
비교예 3. 감귤 과피Comparative Example 3. Citrus peel
상기 실시예 1과 동일하게 실시하되, 감귤 꽃 대신 감귤 과피를 사용하여 천연 식품향료를 수득하였다.Natural food flavors were obtained in the same manner as in Example 1, except that citrus peels were used instead of citrus flowers.
<시험예><Test Example>
시험예 1. 향기성분 분석Test Example 1. Fragrance component analysis
실시예 및 비교예에 따라 제조된 천연 식품향료에 함유된 향기성분은 polydimethyl siloxane(PDMS) 섬유가 장착된 solid phase microextraction(SPME) 장치에 흡착시킨 다음, 가스 크로마토그래피-질량 분석기(gas chromatography-mass spectrometry, GC-MS)로 분석하였다. 분석의 정확도를 위하여 일본 향료 분석 전문 기관에 의뢰하여 향료 Library의 질량 스펙트럼 데이터(Mass Spectrum Data)를 이용하였다. The fragrance components contained in the natural food flavors prepared according to Examples and Comparative Examples were adsorbed on a solid phase microextraction (SPME) device equipped with polydimethyl siloxane (PDMS) fibers, and then gas chromatography-mass spectrometry (gas chromatography-mass spectrometry) It was analyzed by spectrometry, GC-MS). For the accuracy of the analysis, Mass Spectrum Data of the fragrance library was used by requesting a Japanese fragrance analysis agency.
구체적으로, 시료 0.1 g을 20 mL Headspace vials(Wheaton, Millville, USA)에 담은 뒤, 전용 screw cap으로 밀봉시키고 추출 전에 내부표준물질인 Ethyl acetate-d8 (180 μg/mL, 20 μL), 1-butanol-d10 (10 μg/mL, 10 μL), 3-octanone (10 μg/mL, 10 μL), trimethylacetaldehyde (10 μg/mL, 10 μL) 용액들을 시료가 담긴 바이알에 첨가하였다. 기기분석을 위해 SPME-Arrow가 장착된 Trace 1310 gas chromatograph와 TSQ 9000 TQMS (Thermo Scientific, Waltham, USA)를 사용하였으며, 시료들은 DB-WAX column (60 m X 0.25 mm i.d., 0.25 μm film thickness, Agilent Technologies)에서 분리되었다. Carrier gas는 constant flow 모드로 He 1.0 mL/min으로 흘려주었으며, GC inlet의 온도는 220 ℃이며, split mode (split ratio; 5 : 1)로 설정하였다. GC oven programming은 45 ℃에서 10분 동안 유지한 후 분당 2 ℃씩 100 ℃까지 올려준 후 분당 3 ℃씩 230 ℃까지 상승시켰다. MS의 transfer line, MSD source 온도는 각각 230, 280 ℃로 유지되었으며, detector mode는 scan mode로, scan range는 30-550 m/z로 설정하였다. Specifically, after putting 0.1 g of the sample in 20 mL headspace vials (Wheaton, Millville, USA), sealing it with a dedicated screw cap, and before extraction, Ethyl acetate-d 8 (180 μg/mL, 20 μL), 1 Solutions of -butanol-d 10 (10 μg/mL, 10 μL), 3-octanone (10 μg/mL, 10 μL), and trimethylacetaldehyde (10 μg/mL, 10 μL) were added to the vial containing the sample. For instrumental analysis, a Trace 1310 gas chromatograph equipped with SPME-Arrow and a TSQ 9000 TQMS (Thermo Scientific, Waltham, USA) were used. Technologies). The carrier gas was flowed at He 1.0 mL/min in constant flow mode, the temperature of the GC inlet was 220 °C, and the split mode (split ratio; 5: 1) was set. GC oven programming was maintained at 45 ℃ for 10 minutes, then raised to 100 ℃ at 2 ℃ per minute, and then raised to 230 ℃ at 3 ℃ per minute. The MS transfer line and MSD source temperatures were maintained at 230 and 280 °C, respectively, the detector mode was set to scan mode, and the scan range was set to 30-550 m/z .
mint, sweet
위 표 1에 나타낸 바와 같이, 본 발명의 실시예 1에 따라 제조된 천연 식품향료는 향기성분 중에서 D-리모넨(D-Limonene), γ-테르피넨(γ-Terpinene), β-오시멘(β-Ocimene), p-사이멘(p-cymene), β-엘레멘(β-elemene), 카요필렌(Caryophyllene), α-휴물렌(α-Humulene), 제르마크렌 D(Germacrene D), α-파네신(α-Farnesene) 및 벤질 니트릴(Benzyl nitrile)의 함량이 높은 것을 확인하였다.As shown in Table 1 above, the natural food flavor prepared according to Example 1 of the present invention contains D-Limonene, γ-Terpinene, and β-Ocymene among the flavor components. -Ocimene), p-cymene, β-elemene, Caryophyllene, α-Humulene, Germacrene D, α - It was confirmed that the contents of α-Farnesene and Benzyl nitrile were high.
또한, 본 발명의 실시예 1에 따라 제조된 천연 식품향료는 비교예 1 내지 3의 향료에 비하여 총 향기성분의 함량이 많으며, 함량이 높은 향기성분의 종류가 상이한 것을 확인하였다. In addition, it was confirmed that the natural food flavoring agent prepared according to Example 1 of the present invention had a higher content of total fragrance components than the flavoring agents of Comparative Examples 1 to 3, and that the types of fragrance components with high content were different.
시험예 2. 관능검사Test Example 2. Sensory test
실시예 및 비교예에서 제조된 천연 식품향료를 밀봉하여 25 ℃에서 보관하면서 소비자 30명에게 향기를 맡게 한 후 9점 척도법으로 관능검사를 실시하여 평균값 구하였으며, 이를 하기 표 2에 나타내었다. The natural food flavorings prepared in Examples and Comparative Examples were sealed and stored at 25 ° C., and 30 consumers were allowed to smell the fragrance, and then a sensory test was performed using a 9-point scale method to obtain an average value, which is shown in Table 2 below.
- 향취: 1점= 매우 나쁘다, 9점= 매우 좋다- Odor: 1 point = very bad, 9 points = very good
위 표 2에 나타낸 바와 같이, 본 발명의 실시예 1에 따라 제조된 천연 식품향료는 1일차 향이 비교예 1 내지 3에 비하여 우수할 뿐만 아니라 30일차에서도 비교예 1 내지 3에 비하여 우수한 것을 확인하였다.또한, 실시예 1의 천연 식품향료는 30일이 지나도 관능성이 적게 저하되었으나 비교예 1 내지 3은 30일이 지나면서 향취가 거의 없어 관능성이 급격히 저하되는 것을 확인하였다.As shown in Table 2 above, it was confirmed that the natural food flavor prepared according to Example 1 of the present invention was superior to Comparative Examples 1 to 3 on the 1st day as well as superior to Comparative Examples 1 to 3 on the 30th day. In addition, it was confirmed that the natural food flavorings of Example 1 showed a small decrease in their sensuality even after 30 days, but Comparative Examples 1 to 3 had almost no fragrance after 30 days, and their sensuality rapidly decreased.
이에, 실시예 1의 천연 식품향료는 향기성분이 장기간 유지되는 것을 확인하였다. Accordingly, it was confirmed that the natural food flavoring agent of Example 1 was maintained for a long period of time.
시험예 3. 미백 효과 측정Test Example 3. Measurement of whitening effect
3-1. 세포 생존율 측정(MTT assay)3-1. Cell viability measurement (MTT assay)
상기 실시예 및 비교예에서 얻은 시료의 MTT 어세이법에 의한 세포 생존율에 대한 효과를 알아보기 위해 기존 문헌에 기재된 방법을 이용하여 하기와 같이 실험을 수행하였다[BAK J, Pyeon H et al., 2018. Beneficial effects of nano-sized bee pollen on testosterone-induced benign prostatic hyperplasia in rodents. Journal of life science. 28(4) 465-471.]In order to examine the effect of the samples obtained in the above Examples and Comparative Examples on cell viability by the MTT assay, experiments were performed as follows using the method described in the existing literature [BAK J, Pyeon H et al., 2018. Beneficial effects of nano-sized bee pollen on testosterone-induced benign prostatic hyperplasia in rodents. Journal of life science. 28(4) 465-471.]
실시예 및 비교예에서 얻은 시료를 125, 250, 500 μg/ml의 농도로 B16F10 세포에 처리하여 세포 생존율을 통한 세포독성을 평가하였다.Samples obtained in Examples and Comparative Examples were treated with B16F10 cells at concentrations of 125, 250, and 500 μg/ml to evaluate cytotoxicity through cell viability.
세포주를 1X104 cells/well로 96 well plate에 180 uL씩 분주하여 24시간 배양하였다. 이후 배지를 제거하고 추출물을 농도별로 희석하여, 20 uL씩 plate에 분주하고 37 ℃의 5% CO2 항온배양기에서 72시간 배양하였다. 배지를 제거하고 MTT 시약을 0.25 mg/mL로 PBS에 용해시켜 배지와 혼합하여 plate에 분주하여 4시간 배양한 후 formazon 결정을 위해 시약을 모두 제거하고 DMSO를 100 uL 분주하여 microplate reader를 이용하여 540 nm에서 흡광도를 측정하였다.The cell line was dispensed at 1X10 4 cells/well by 180 uL into a 96 well plate and cultured for 24 hours. Thereafter, the medium was removed, and the extract was diluted by concentration, dispensed on a plate by 20 uL, and cultured for 72 hours in a 5% CO 2 incubator at 37 °C. Remove the medium, dissolve the MTT reagent at 0.25 mg/mL in PBS, mix it with the medium, distribute it to the plate, and incubate for 4 hours. Remove all reagents for formazon determination, dispense 100 uL of DMSO, and use a microplate reader at 540 °C. Absorbance was measured in nm.
위 표 3에 나타낸 바와 같이, 본 발명의 실시예 1 및 비교예 1 내지 3에 따라 제조된 천연 항료는 세포독성이 없는 것으로 확인되었다.As shown in Table 3 above, it was confirmed that the natural flavors prepared according to Example 1 and Comparative Examples 1 to 3 of the present invention were not cytotoxic.
3-2. B16F10세포에 대한 멜라닌 합성 저해 효과 측정3-2. Measurement of melanin synthesis inhibitory effect on B16F10 cells
실시예 및 비교예에서 제조된 천연 항료가 멜라닌 합성에 미치는 영향을 알아보기 위하여 B16F10 세포에 천연 항료를 농도별로 처리하고 세포 내 멜라닌 함량을 측정하였다. 보다 자세하게는 B16F10세포를 12 well plate에 well당 1 X 105 세포로 분주한 후 24시간 뒤 천연 항료를 농도별로 1시간 동안 전처리하고, α-MSH 1 μM로 72시간 동안 처리한 다음 72시간 뒤에 PBS로 2번 세척하였다. 세척된 세포는 1N NaOH로 세포를 파쇄한 후 EP 튜브에 담아서 30분간 60 ℃에서 가열하였다. 세포 용해물을 5000 xg에서 5분간 원심분리한 후 상층액의 흡광도를 405 nm에서 측정하였다. In order to examine the effect of the natural fragrance prepared in Examples and Comparative Examples on melanin synthesis, B16F10 cells were treated with the natural fragrance at each concentration and the intracellular melanin content was measured. More specifically, B16F10 cells were divided into 1 X 10 5 cells per well in a 12-well plate, 24 hours later, pre-treatment with natural dyes for 1 hour at each concentration, treatment with α-MSH 1 μM for 72 hours, and 72 hours later Washed twice with PBS. The washed cells were lysed with 1N NaOH, placed in an EP tube, and heated at 60 °C for 30 minutes. After centrifuging the cell lysate at 5000 xg for 5 minutes, the absorbance of the supernatant was measured at 405 nm.
위 표 4에 나타낸 바와 같이, 본 발명의 실시예 1에 따라 제조된 천연 식품향료는 비교예 1 내지 3에 비하여 B16F10세포에서 멜라닌 합성을 억제하는 것을 확인하였다. As shown in Table 4 above, it was confirmed that the natural food flavor prepared according to Example 1 of the present invention inhibits melanin synthesis in B16F10 cells compared to Comparative Examples 1 to 3.
3-3. B16F10세포에 대한 티로시나아제(Tyrosinase) 활성 저해 효과 측정3-3. Measurement of tyrosinase activity inhibitory effect on B16F10 cells
티로시나아제 활성 저해능은 Flurkey의 방법을 변형하여 실험하였다. 증류수 100 mL에 각각 sodium phosphate monobasic anhydrous 0.8039 g, sodium phosphate dibasic anhydrous 0.9511 g을 녹여 pH를 6.8로 조정하여 buffer를 제조하였다. 제조된 buffer를 용매로 기질 10 mM 3,4-dihydroxy phenylanin(I-dopa) (20 mg/mL)와 효소 1250 unit tyrosinase를 제조하였으며 증류수를 혼합하여 1250 unit tyrosinase를 10배 희석하여 사용하였다. 기질 0.2 g, 시료 0.2 g, 효소 0.2 g을 순서대로 첨가하여 교반한 뒤 25 ℃에서 30분간 반응시켰다. 기질 0.2 g, 시료 0.2 g, 효소 0.2 g을 마이크로 튜브에 순서대로 첨가하여 교반한 뒤 25 ℃에서 30분간 반응시켰다. 반응 후 475 nm의 파장에서 흡광도를 측정하였다. The ability to inhibit tyrosinase activity was tested by modifying Flurkey's method. A buffer was prepared by dissolving 0.8039 g of sodium phosphate monobasic anhydrous and 0.9511 g of sodium phosphate dibasic anhydrous in 100 mL of distilled water and adjusting the pH to 6.8. Substrate 10 mM 3,4-dihydroxy phenylanin (I-dopa) (20 mg/mL) and enzyme 1250 unit tyrosinase were prepared using the prepared buffer as a solvent, and 1250 unit tyrosinase was diluted 10 times with distilled water. 0.2 g of substrate, 0.2 g of sample, and 0.2 g of enzyme were sequentially added and stirred, followed by reaction at 25 °C for 30 minutes. 0.2 g of substrate, 0.2 g of sample, and 0.2 g of enzyme were sequentially added to the microtube, stirred, and reacted at 25 °C for 30 minutes. After the reaction, absorbance was measured at a wavelength of 475 nm.
위 표 5에 나타낸 바와 같이, 본 발명의 실시예 1에 따라 제조된 천연 식품향료는 비교예 1 내지 3에 비하여 B16F10세포에서 티로시아나제 생성을 억제하는 것을 확인하였다. As shown in Table 5 above, it was confirmed that the natural food flavor prepared according to Example 1 of the present invention inhibits tyrosyanase production in B16F10 cells compared to Comparative Examples 1 to 3.
3-4. 제브라피쉬를 이용한 멜라닌 합성3-4. Melanin synthesis using zebrafish 저해 효과 측정Inhibition effect measurement
제브라피쉬(zebrafish)에서 멜라닌 합성 저해능을 평가하였다. 제브라 피쉬 10 마리의 물고기를 28.5 ℃ 3 L 아크릴 탱크에 14:10 시간의 명암 주기로 사육하였으며, 제브라 피쉬는 주 6일 동안 하루에 두 번 Tetramin 플레이크 식품을 보충했습니다. 배아를 채집하여 배아 발달 단계에서 멜라닌 생성에 천연 식품향료가 미치는 영향을 조사하기 위하여 α-MSH 자극제를 사용하였다. 제브라 피쉬 배아 (7-9 hpf) 30개를 6 well 플레이트에서 배아 배지 1.9 mL 분주하였다. 천연 식품향료를 PBS와 함께 1 % DMSO에 용해하여 혼합하여 처리하였다. 멜라닌 함량을 측정하기 위하여 3일 후 잔류 시약 또는 입자를 제거하기 위하여 제브라 피쉬를 PBS로 2회 세척 후에 현미경으로 사진을 측정하고 e-tube에 넣어 원심 분리 후, 펠렛을 90 ℃에서 1 mL의 1N NaOH에 60 분 동안 교반하며 용해시켰다. 그리고 96 well 플레이트에 200 uL 분주하여 490 nm에 흡광도를 측정하여 계산하였다. The ability to inhibit melanin synthesis was evaluated in zebrafish. Zebrafish Ten fish were reared in a 3 L acrylic tank at 28.5 °C with a 14:10 hour light/dark cycle, and the zebrafish were supplemented with Tetramin flake food twice a day for 6 days a week. Embryos were collected and α-MSH stimulants were used to investigate the effect of natural food flavors on melanin production during embryonic development. 30 zebrafish embryos (7-9 hpf) were dispensed in 1.9 mL of embryo medium in a 6-well plate. Natural food flavors were dissolved in 1% DMSO with PBS and mixed to treat. After 3 days to measure the melanin content, the zebrafish were washed twice with PBS to remove residual reagents or particles, and then the pictures were taken under a microscope, placed in an e-tube, centrifuged, and pelleted at 90 ° C in 1 mL of 1N. Dissolved in NaOH with stirring for 60 min. Then, 200 uL was dispensed into a 96 well plate, and the absorbance was measured at 490 nm and calculated.
도 1A는 본 발명의 실시예 1에 따라 제조된 천연 식품향료가 처리된 제브라피쉬를 멜라닌 생성 억제를 나타낸 사진이며, 도 1B는 상기 도 1A의 제브라피쉬에 생성된 멜라닌을 정량화한 그래프이다.Figure 1A is a photograph showing the inhibition of melanin production in zebrafish treated with a natural food flavor prepared according to Example 1 of the present invention, Figure 1B is a graph quantifying melanin produced in the zebrafish of Figure 1A.
도 1A 및 도 1B에 도시된 바와 같이, 시료를 처리하지 않은 대조군(αMSH)에 비해 100 내지 400 ㎍/ml의 실시예 1 천연 식품향료를 처리한 제브라피쉬 배아의 요크(yolk) 부분에서 멜라닌 색소의 침착이 저해될 뿐만 아니라 전체적으로 멜라닌의 합성이 저해됨을 확인하였다.As shown in FIGS. 1A and 1B, melanin pigment in the yolk portion of zebrafish embryos treated with 100 to 400 μg/ml of Example 1 natural food flavor compared to the control group (αMSH) not treated with the sample. It was confirmed that not only the deposition of but also the inhibition of melanin synthesis as a whole.
하기에 본 발명의 추출물을 함유하는 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, formulation examples of the composition containing the extract of the present invention are described, but the present invention is not intended to limit them, but is intended to be specifically described.
제제예 1. 과립제의 제조Formulation Example 1. Preparation of granules
실시예 1에서 얻은 천연 식품향료 1,000 mg1,000 mg of natural food flavor obtained in Example 1
비타민 혼합물 적량Appropriate amount of vitamin mixture
비타민 A 아세테이트 70 ㎍Vitamin A Acetate 70 μg
비타민 E 1.0 mgVitamin E 1.0 mg
비타민 B1 0.13 mgVitamin B1 0.13 mg
비타민 B2 0.15 mgVitamin B2 0.15 mg
비타민 B6 0.5 mgVitamin B6 0.5 mg
비타민 B12 0.2 ㎍Vitamin B12 0.2 μg
비타민 C 10 mgVitamin C 10 mg
비오틴 10 ㎍10 μg of biotin
니코틴산아미드 1.7 mgNicotinamide 1.7 mg
엽산 50 ㎍Folic acid 50 μg
판토텐산 칼슘 0.5 mgCalcium Pantothenate 0.5 mg
무기질 혼합물 적량Appropriate amount of mineral mixture
황산제1철 1.75 mgFerrous sulfate 1.75 mg
산화아연 0.82 mgZinc Oxide 0.82 mg
탄산마그네슘 25.3 mgMagnesium Carbonate 25.3 mg
제1인산칼륨 15 mgPotassium Phosphate Monobasic 15 mg
제2인산칼슘 55 mgDibasic Calcium Phosphate 55 mg
구연산칼륨 90 mg
탄산칼슘 100 mgCalcium Carbonate 100 mg
염화마그네슘 24.8 mgMagnesium Chloride 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 과립제에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 과립제 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강기능식품 조성물 제조에 사용할 수 있다.Although the composition ratio of the above vitamin and mineral mixture was prepared by mixing ingredients suitable for granules in a preferred embodiment, the mixing ratio may be arbitrarily modified, and after mixing the above ingredients according to a conventional granule manufacturing method, It can be prepared and used for preparing a health functional food composition according to a conventional method.
제제예 2. 기능성 음료의 제조Formulation Example 2. Manufacturing of functional beverages
실시예 1에서 얻은 천연 식품향료 1,000 mg1,000 mg of natural food flavor obtained in Example 1
구연산 1,000 mgCitric Acid 1,000 mg
올리고당 100 g100 g of oligosaccharides
매실농축액 2 g2 g plum concentrate
타우린 1 g1 g of taurine
정제수를 가하여 전체 900 mLAdd purified water to total 900 mL
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1 시간 동안 85 ℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 L 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 기능성 음료 조성물 제조에 사용한다. After mixing the above ingredients according to the normal health drink manufacturing method, stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered and collected in a sterilized 2 L container, sealed and sterilized, and then refrigerated. It is used for preparing the functional beverage composition of the present invention.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the composition ratio is a mixture of ingredients suitable for a relatively favorite beverage in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the class of demand, the country of demand, and the purpose of use.
본 발명을 적용하기에 적합한 화장료 조성물의 제조예를 제시하기로 한다.Preparation examples of cosmetic compositions suitable for application of the present invention will be presented.
제제예 3: 화장수Formulation Example 3: Lotion
실시예 1에서 얻은 천연 식품향료를 포함하는 화장료 중 화장수의 제조예는 하기 표 6과 같다.Table 6 shows examples of preparation of lotion among cosmetics containing the natural food flavor obtained in Example 1.
제제예 4: 로션Formulation Example 4: Lotion
실시예 1에서 얻은 천연 식품향료를 포함하는 화장료 중 로션의 제조예는 하기 표 7과 같다.Examples of preparation of the lotion among cosmetics containing the natural food flavor obtained in Example 1 are shown in Table 7 below.
제제예 5: 영양 크림Formulation Example 5: Nutrition Cream
실시예 1에서 얻은 천연 식품향료를 포함하는 화장료 중 영양 크림의 제조예는 하기 표 8과 같다.Table 8 shows examples of the preparation of nutritious cream among cosmetics containing natural food flavorings obtained in Example 1.
제제예 6: 에센스Formulation Example 6: Essence
실시예 1에서 얻은 천연 식품향료를 포함하는 화장료 중 에센스의 제조예는 하기 표 9와 같다.Table 9 shows examples of the preparation of essences among cosmetics containing natural food flavorings obtained in Example 1.
제제예 7: 마스크 팩용 유액Formulation Example 7: Emulsion for mask pack
실시예 1에서 얻은 천연 식품향료를 포함하는 화장료 중 마스크 팩용 유액의 제조예는 하기 표 10과 같다.Table 10 shows examples of preparation of emulsions for mask packs among cosmetics containing natural food flavors obtained in Example 1.
Claims (14)
(B) 상기 동결건조된 꽃을 분쇄한 후 30 내지 60 ℃에서 용매로 1차 추출하여 에센셜 오일을 수득하는 단계; 및
(C) 상기 1차 추출된 에센셜 오일을 2차로 초임계 추출하여 천연 식품향료를 수득하는 단계;를 포함하는 것을 특징으로 하는 천연 식품향료의 제조방법.(A) freeze-drying the flowers;
(B) obtaining essential oil by first extracting the freeze-dried flowers with a solvent at 30 to 60 ° C.; and
(C) obtaining a natural food flavor by performing second supercritical extraction of the first extracted essential oil.
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