KR102247944B1 - Antioxidant and whitening composition - Google Patents

Antioxidant and whitening composition Download PDF

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KR102247944B1
KR102247944B1 KR1020190056913A KR20190056913A KR102247944B1 KR 102247944 B1 KR102247944 B1 KR 102247944B1 KR 1020190056913 A KR1020190056913 A KR 1020190056913A KR 20190056913 A KR20190056913 A KR 20190056913A KR 102247944 B1 KR102247944 B1 KR 102247944B1
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glutathione
antioxidant
plum
composition
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KR20200133047A (en
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김혁주
차재윤
봉하균
김선일
이성태
장경
박성진
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순천대학교 산학협력단
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    • AHUMAN NECESSITIES
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Abstract

기존의 피부 미백 조성물은 피부에 바르는 물질이 대부분이다. 그러나 피부에 바르는 미백 조성물은 사용 시기가 한정되어 있고, 효과도 불분명하였다. 이러한 문제를 해결하기 위하여, 본 발명의 항산화 및 미백 조성물은 매실 5 중량%, 자일리톨 50 중량%, 결정과당 29.5 중량%, 비타민 C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 0.5 중량% 인 것을 특징으로 하는 항산화 및 미백 조성물을 제공한다.
상기와 같은 구성에 의하여 매실의 항산화 기능과 글루타치온 효모의 활성산소 제거 기능을 이용한 식품조성물을 제공함으로서 건강한 생활을 할 수 있는 조성물을 제공한다.
Most of the existing skin whitening compositions are substances applied to the skin. However, the time of use of the whitening composition applied to the skin was limited, and the effect was unclear. In order to solve this problem, the antioxidant and whitening composition of the present invention is glutathione containing 5% by weight of plum, 50% by weight of xylitol, 29.5% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 50% by weight of glutathione. It provides an antioxidant and whitening composition, characterized in that 0.5% by weight of yeast.
By providing a food composition using the antioxidant function of plum and the active oxygen removal function of glutathione yeast by the above configuration, a composition capable of living a healthy life is provided.

Description

항산화 및 미백 조성물{.}Antioxidant and whitening composition{.}

본 발명은 항산화 및 미백 기능이 있는 식품조성물에 관한 것으로 더욱 자세하게는 Glutathione Yeast를 함유하는 매실추출물에 관한 것이다.The present invention relates to a food composition having antioxidant and whitening functions, and more particularly, to a plum extract containing Glutathione Yeast.

본 발명 이전의 선행기술로는 홍화황 추출물을 포함하는 미백용 화장료 조성물, 미백용 식품 조성물, 상기 조성물을 포함하는 색소침착 질환의 예방 또는 완화용 건강기능성 식품, 및 색소 침착 질환의 예방 또는 치료용 약제학적 조성물에 관한 기술이 개시되어 있다.Prior art prior to the present invention includes a cosmetic composition for whitening, a food composition for whitening, a health functional food for the prevention or alleviation of pigmentation diseases containing the composition, and for the prevention or treatment of pigmentation diseases Description of the pharmaceutical composition is disclosed.

또 다른 선행기술로는 노근, 함초 및 매실 추출물을 함유하는 항산화 및 미백 화장료 조성물에 관한 것으로서, 노근의 에탄올 추출 동결 건조물에 대한 에틸아세테이트 추출 분획물과, 함초의 에탄올 추출 동결 건조물에 대한 에틸아세테이트 또는 부탄올 추출 분획물과, 매실의 에탄올 추출 동결 건조물의 혼합물을 포함하는 구성이 개시되어 있다.Another prior art relates to an antioxidant and whitening cosmetic composition containing extracts of nogeun, green tea and plum, and an ethyl acetate-extracted fraction for the ethanol-extracted freeze-dried product of Nogeun, and ethyl acetate or butanol for the freeze-dried product of ethanol-extracted green tea A composition comprising a mixture of an extracted fraction and a freeze-dried product of ethanol extraction of plum is disclosed.

등록특허공보 10-1768626Registered Patent Publication 10-1768626 등록특허공보 10-1704874Registered Patent Publication 10-1704874

기존의 피부 미백 조성물은 피부에 바르는 물질이 대부분이다. 그러나 피부에 바르는 미백 조성물은 사용시기가 한정되어 있고, 효과도 불분명하였다. 이러한 문제를 해결하기 위하여, 본 발명은 먹는 항산화 및 미백 조성물을 제공하고자한다. 또한, 본 발명은 매실의 항산화 기능과 글루타치온 효모의 활성산소 제거기능을 이용한 식품조성물을 제공한다. Most of the existing skin whitening compositions are substances applied to the skin. However, the time of use of the whitening composition applied to the skin was limited, and the effect was unclear. In order to solve this problem, the present invention is to provide an antioxidant and whitening composition for eating. In addition, the present invention provides a food composition using the antioxidant function of plum and the active oxygen removal function of glutathione yeast.

본 발명은 기존의 문제를 해결하기 위하여 하기의 과제 해결 수단을 제공한다.The present invention provides a means for solving the following problems in order to solve the existing problems.

본 발명의 항산화 및 미백 조성물은 매실 5 중량%, 자일리톨 50 중량%, 결정과당 29.5 중량%, 비타민 C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 0.5 중량% 인 것을 특징으로 하는 항산화 및 미백 조성물을 제공한다.The antioxidant and whitening composition of the present invention is 0.5% by weight of glutathione yeast containing 5% by weight of plum, 50% by weight of xylitol, 29.5% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 0.5% by weight of glutathione. It provides an antioxidant and whitening composition.

또 다른 실시예로, 본 발명의 항산화 및 미백 조성물은 매실 5 중량%, 자일리톨 50 중량%, 결정과당 29 중량%, 비타민 C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 1 중량% 인 것을 특징으로 하는 항산화 및 미백 조성물을 제공한다.In another embodiment, the antioxidant and whitening composition of the present invention is a glutathione yeast containing 5% by weight of plum, 50% by weight of xylitol, 29% by weight of fructose, 7% of vitamin C, 3% by weight of minerals, and 50% by weight of glutathione. It provides an antioxidant and whitening composition, characterized in that 1% by weight.

또 다른 실시예로, 본 발명의 항산화 및 미백 조성물은 매실 5 중량%, 자일리톨 50 중량%, 결정과당 28.5 중량%, 비타민 C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 1.5 중량% 인 것을 특징으로 하는 항산화 및 미백 조성물을 제공한다.In another embodiment, the antioxidant and whitening composition of the present invention is a glutathione yeast containing 5% by weight of plum, 50% by weight of xylitol, 28.5% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 50% by weight of glutathione. It provides an antioxidant and whitening composition, characterized in that 1.5% by weight.

본 발명은 상기와 같은 구성에 의하여 항산화 실험 결과 Fe

Figure 112019049653517-pat00001
chelating 능력을 측정한 결과 Glutathione 효모를 함유한 실험군에서 유의한 저해 효과를 나타내었다. 또한 본 발명의 멜라닌 생합성의 경우에도 X50배를 희석한 Glutathione 효모를 함유한 실험군에서 통계적으로 유의하게 더욱 생합성을 저해하는 효과를 나타내었다. 또한 글루타치온 효모를 이취 없이 섭취하기 위하여 HPMC를 첨가하여 과립화하는 과정에서 항산화 활성과 미백효과는 영향을 받지 않았다.The present invention is a result of the antioxidant experiment by the configuration as described above Fe
Figure 112019049653517-pat00001
As a result of measuring the chelating ability, a significant inhibitory effect was shown in the experimental group containing Glutathione yeast. In addition, in the case of the melanin biosynthesis of the present invention, the experimental group containing the X50-fold diluted Glutathione yeast showed a statistically significant effect of further inhibiting biosynthesis. In addition, the antioxidant activity and whitening effect were not affected in the process of granulating by adding HPMC in order to consume glutathione yeast without off-flavor.

또한, 본 발명은 매실의 항산화 기능과 글루타치온 효모의 활성산소 제거 기능을 이용한 식품조성물을 제공함으로서 건강한 생활을 할 수 있는 조성물을 제공한다. In addition, the present invention provides a composition capable of living a healthy life by providing a food composition using the antioxidant function of plum and the function of removing active oxygen from glutathione yeast.

도1은 Fe chelating ability
도 2는 Tyrosinase inhibition rate
도3은 Cytotoxicity of extract on B16F0 cells
도4는 세포 내 tyrosinase 활성 저해 효과
도5는 멜라닌 생합성 저해 효과
도6은 항산화 활성 측정
도7은 The ABTS+ cation radical scavenging activity of containing HPMC products and except HPMC product
도8은 Frap The Fe3+-TPTZ-Fe2+-TPTZ reducing ability of containing HPMC products and except HPMC product
도9는 The Fe2+ chelating of different of containing HPMC products and except HPMC product and except HPMC product
도10은 Tyrosinase(from mushroom) inhibition rate of containing HPMC products and except HPMC product
Figure 1 is Fe chelating ability
Figure 2 is the Tyrosinase inhibition rate
Figure 3 is a Cytotoxicity of extract on B16F0 cells
Figure 4 shows the effect of inhibiting tyrosinase activity in cells
5 is a melanin biosynthesis inhibitory effect
Figure 6 is an antioxidant activity measurement
7 shows The ABTS+ cation radical scavenging activity of containing HPMC products and except HPMC product.
Figure 8 shows Frap The Fe3+-TPTZ-Fe2+-TPTZ reducing ability of containing HPMC products and except HPMC product.
9 shows The Fe2+ chelating of different of containing HPMC products and except HPMC product and except HPMC product.
Figure 10 is Tyrosinase (from mushroom) inhibition rate of containing HPMC products and except HPMC product

최근 생활과 소득 수준이 향상되면서 삶의 질을 향상시키기 위해 많은 관심이 높아지고 있다. 특히 노화와 각종 염증질환, 암의 대처 방안으로 항산화 물질을 비롯한 체내 질병과 대사를 위한 생리활성물질 개발에 대해 관심이 증대 되고 있다. 생체 내에 필요한 에너지 공급을 위해 생화학적 산화 반응은 끊임없이 일어나며 이 과정 중에 발생하는 유해산소라 불리는 활성산소 (reactive oxygen species, ROS) 는 가장 안정한 형태의 산소인 삼중항산소가 산화, 환원과정에서 생성되는 일중항산소인 Superoxide anion, hydroxyl radical, hydrogen peroxide과 같은 불안정한 상태의 프리라디칼(free radical) 및 과산화수소 (H2O2)로 불안정하고 산화력이 높아 생체물질과 쉽게 반응하기 때문에 인체 내에서 제거되지 못하면 산화적 스트레스 (oxidative stress)를 유발하게 되며, 이러한 산화적 스트레스 중 피부 광 손상의 중요한 인자인 O2 및 OH는 피부에 존재하는 항산화 물질의 파괴, 지질과산화반응의 개시, 단백질 및 DNA 산화, 결합조직 성분인 콜라겐(collagen)과 엘라스틴(elastin) 및 히알루 론산(hyaluronic acid) 등의 결합사슬 절단 및 비정상적인 교차결합에 의한 주름생성, 멜라닌 생성촉진 등 피부의 노화를 가속화 한다.Recently, as the level of living and income has improved, a lot of interest is increasing to improve the quality of life. In particular, interest in the development of physiologically active substances for metabolism and diseases in the body, including antioxidants, is increasing as a countermeasure against aging, various inflammatory diseases, and cancer. Biochemical oxidation reactions constantly occur in order to supply energy necessary for the living body, and reactive oxygen species (ROS), which is called harmful oxygen, generated during this process is the most stable form of oxygen, triplet oxygen, which is produced during the oxidation and reduction process. It is unstable with unstable free radicals and hydrogen peroxide (H2O2) such as singlet oxygen superoxide anion, hydroxyl radical, and hydrogen peroxide. (oxidative stress), and among these oxidative stresses, O 2 and OH, which are important factors of skin light damage, destroy antioxidants present in the skin, initiate lipid peroxidation, oxidation of proteins and DNA, and connective tissue components. It accelerates skin aging such as cleavage of binding chains such as collagen, elastin, and hyaluronic acid, wrinkle formation due to abnormal cross-linking, and promotion of melanin production.

피부는 나이가 들어감에 따라 자연스럽게 생기는 내인성노화 (Intrinisc againg)와 누적된 햇빛의 노출에 의해 유발되는 광노화 (Photoaging)가 있다. 이 중 광노화는 피부노화의 주된 원인으로 최근 급격하게 변화하는 산업화와 환경오염으로 인해 오존층이 파괴되어 자외선 조사량이 심각하게 방출됨으로써 증가하고 있는 추세이다. 자외선 (Ultravoiler. UV)은 태양에서 발생되는 400nm 이하의 파장을 가지는 광선으로 생체에 많은 변화를 가져온다. 이는 진피의 유두층, 그물층까지 영향을 미치고 탄력소와 아교질의 붕괴로 탄력감소, 조기노화, 모세혈관의 확장 및 손상으로 피부의 기저층을 와해시키며, 피부암 발생 가능성도 높아진다. 모발 역시 태양광선에 노출되면 건조해지고, 강도가 감소되며, 표면이 거칠어지고 쉽게 부서지게 된다. 이렇게 태양광선에 의한 광화학적 분해는 모발이나 피부 단백질과 멜라닌에 자극을 주어 내부세포, 세포막 혼합체, 멜라닌의 파괴가 일어나게 된다. 멜라닌은 검은 색소와 단백질의 복합체이며 사람의 머리카락과 피부색을 이루는 색소로서 양이 많으면 피부색이 황갈색에서 흑갈색을 띄고 양이 적을수록 색이 엷어지게 된다. 멜라닌의 경우 외부환경, 자외선으로부터 피부세포를 보호하는 역할을 하지만 과도한 자외선 노출이나 피부노화로 인해 과잉생성 될 경우 피부에 색소가 침착되어 기미, 주근깨를 형성하며 멜라닌 전구물질의 독성으로 피부암의 원인이 되기도 한다. 따라서 이러한 색소 침착현상을 방지하기 위해서는 멜라닌 생성과정의 일부분을 저해하여 멜라닌의 생성을 감소시켜야 한다. 멜라닌의 생합성에서 가장 중요한 단계는 타이로시네이즈 (tyrosinase) 의 촉매작용을 통하여 일어나는 초기 반응으로 표피의 기저층에 존재하는 melanocyte (멜라노사이트) 내의 melanosome (멜라노좀) 이라는 소포체에서 먼저 tyrosinase, tyrosinase related protein-1(TRP-1)과 dopachrome tautomerase (DCT)등의 효소에 의해 생성된다.The skin has intrinsc againg, which occurs naturally with age, and photoaging, which is caused by accumulated sunlight exposure. Among them, photoaging is a major cause of skin aging, and it is increasing as the ozone layer is destroyed due to industrialization and environmental pollution, which are rapidly changing in recent years, and the amount of ultraviolet irradiation is seriously released. Ultraviolet (Ultravoiler. UV) is a ray of light with a wavelength of 400 nm or less generated from the sun and causes many changes in the living body. This affects the papillary layer and the reticulum layer of the dermis, and the base layer of the skin is damaged by loss of elasticity, premature aging, expansion and damage of capillaries due to the collapse of elasticity and collagen, and the likelihood of skin cancer is increased. Hair, too, becomes dry when exposed to sunlight, its strength decreases, and its surface becomes rough and brittle. In this way, photochemical decomposition by sunlight stimulates hair or skin proteins and melanin, resulting in destruction of internal cells, cell membrane mixtures, and melanin. Melanin is a complex of black pigment and protein, and it is a pigment that makes up the color of human hair and skin. When the amount is high, the skin color becomes yellowish brown to blackish brown, and the smaller the amount, the lighter color. In the case of melanin, it protects skin cells from the external environment and ultraviolet rays. However, when over-produced due to excessive exposure to ultraviolet rays or skin aging, pigmentation is deposited on the skin to form spots and freckles, and the toxicity of melanin precursors causes skin cancer. It is also possible. Therefore, in order to prevent such pigmentation phenomenon, it is necessary to reduce the production of melanin by inhibiting a part of the melanin production process. The most important step in the biosynthesis of melanin is the initial reaction that occurs through the catalytic action of tyrosinase.In the endoplasmic reticulum called melanosome (melanosome) in the base layer of the epidermis, tyrosinase, tyrosinase related protein It is produced by enzymes such as -1 (TRP-1) and dopachrome tautomerase (DCT).

앞서 언급된 타이로시네이즈(tyrosinase)는 멜라닌 생성 첫 단계를 일으키는 중요한 효소로, tyrosin (타이로신)이 3, 4-dihydroxy phenylalanine (DOPA, 도파)를 거쳐 DOPAquinone (도파퀴논)으로 전환되고, DOPAquinone 으로부터 자동 산화 반응과 효소 반응으로 DOPAchrome (도파크롬)을 거쳐 흑갈색의 공중합체인 멜라닌을 생성하게 된다. The aforementioned tyrosinase is an important enzyme that causes the first stage of melanogenesis, and tyrosin (tyrosine) is converted to DOPAquinone (dopaquinone) through 3,4-dihydroxy phenylalanine (DOPA, dopa), and from DOPAquinone. It produces melanin, a dark brown copolymer, through DOPAchrome (Dopachrome) through an automatic oxidation reaction and an enzymatic reaction.

현재 과도하게 생성된 멜라닌의 부정적인 기능 때문에 미백제의 개발에 있어서, 멜라닌 생합성 대사에 대한 연구가 진행 중이며 그 중에서도 멜라닌 생성의 주요 효소인 타이로시네이즈 (tyrosinase) 활성을 저해함으로써 멜라닌 생성을 억제시켜 미백효과를 유도할 수 있다.Currently, due to the negative function of excessively produced melanin, studies on the metabolism of melanin biosynthesis are underway in the development of whitening agents. Among them, whitening by inhibiting melanin production by inhibiting the activity of tyrosinase, a major enzyme in melanogenesis. It can induce an effect.

기존 화장품 분야에서는 미백성분으로서, tyrosinase 효소활성을 억제하는 물질로 알려진 코지산 (kojic acid), 알부틴(arbutin) 을 비롯한 하이드로퀴논 (hydroquinone), 비타민C 및 이들의 유도체와 각종 식물 추출물이 사용되어 왔다. 그러나, 이런 물질들은 불안정하여 분해나 착색, 이취, 효과의 불분명, 피부와 제형 안전성 문제뿐만 아니라 돌연변이 유발 및 세포독성문제 등으로 그 사용이 제한되고 있는 실정이다. 따라서 기존 미백제가 갖고 있는 여러 단점을 극복하고자 최근에는 피부에 안전한 천연물을 이용한 미백제 개발에 많은 연구가 진행되고 있다.In the existing cosmetics field, as whitening ingredients, hydroquinones, including kojic acid and arbutin, which are known to inhibit tyrosinase enzyme activity, vitamin C, derivatives thereof, and various plant extracts have been used. . However, these substances are unstable, and their use is limited due to decomposition, coloration, off-flavor, unclear effect, safety problems of skin and formulation, as well as mutagenesis and cytotoxicity problems. Therefore, in order to overcome the various disadvantages of existing whitening agents, many studies are being conducted on the development of whitening agents using natural products that are safe for the skin.

매실나무(Prunus mume)는 장미과에 속하며 원산지는 중국의 동남부지방이며 한국, 중국 및 일본의 온난한 지역에 분포하는 동양 고유종의 과수 이다. 또한 나관중의 ‘삼국지연의’에 나올 정도로 옛날부터 식용 또는 약용으로 사용되어 왔고 각종 한의서에 매실은 만성기침, 위염, 만성사, 혈뇨, 구토, 치질등 치료한 기록이 전해진다. 매실은 malic acid, citric acid, oxalic acid, succinic acid, fumaric acid 와 같은 유기산과 무기질을 많이 포함하고 있어 식욕촉진 및 위액분비를 증가하여 소화 활동과 피로회복을 도와주는 효과가 있다고 알려져 있다. 그 중에서도 매실의 중요한 성분 중에 하나인 루틴(rutin)은 혈관계 질환의 치료와 모세혈관 강화, 항염증 효과에 도움을 주며, 항산화 효능을 발휘한다고 보고되고 있고 항산화 효능을 이용한 미백효능에 대한 연구도 진행되었다.Plum tree (Prunus mume) belongs to the Rosaceae family, and its origin is in the southeastern part of China, and it is an endemic fruit tree of the East distributed in the warm regions of Korea, China and Japan. In addition, it has been used for edible or medicinal purposes since ancient times to the extent that it appeared in Nagwanjung’s “Three Kingdoms”. In various oriental medicine books, there are records of treatment of plums such as chronic cough, gastritis, chronic death, hematuria, vomiting, and hemorrhoids. Plums contain a lot of organic acids and minerals such as malic acid, citric acid, oxalic acid, succinic acid, and fumaric acid, and are known to have the effect of promoting appetite and increasing gastric juice secretion to help digestive activity and fatigue recovery. Among them, rutin, one of the important components of plum, helps treat vascular disease, strengthens capillaries, and has anti-inflammatory effects, and is reported to exhibit antioxidant effects, and studies on whitening efficacy using antioxidant effects are also underway. Became.

본 발명에서는 매실과 글루타티온(Glutathione)을 함유하는 효모 혼합물이 항산화 및 미백 효과를 가지는 것을 확인하였고 이를 활용하고자 한다.In the present invention, it was confirmed that a yeast mixture containing plum and glutathione has antioxidant and whitening effects, and it is intended to be utilized.

이를 위하여 항산화 실험과 멜라닌 생합성 실험 및 이취 제거 및 과립화를 위하여 첨가한 HPMC에 의한 상기 항산화 활성과 미백효과의 영향 정도를 측정하는 실험을 하였다.To this end, an experiment was performed to measure the degree of influence of the antioxidant activity and whitening effect by HPMC added for the antioxidant experiment, melanin biosynthesis experiment, off-flavor removal and granulation.

실험방법Experiment method

1) 재료 및 시약1) Materials and reagents

DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diamonium salt), Iron (Ⅲ) chloride, Potassium ferricyanide, Trichloroacetic acid, Iron(Ⅱ) chloride, 2,2’-Bipyridyl, Copper (Ⅱ) chloride, Ammonium acetate, Pyrogallol, Quercetin, Folin & Ciocalteu’s phenol reagent, Gallic acid, Vanillin, Catechin, TPTZ (2,4,6-tripyridyl-S-triazine), Ferric Chloride는 Sigma-Aldrich사 (St. Louis, Mo, USA)에서 각각 구입하여 사용하였다. Glacial acetic acid 는 Alfa Aesar(Thermo Fisher Scientific)에서 구입하여 사용하였으며 Sodium acetate trihydrate는 JUNSEI 에서 구입하여 사용하였다.DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diamonium salt), Iron (Ⅲ) chloride, Potassium ferricyanide, Trichloroacetic acid, Iron( Ⅱ) chloride, 2,2'-Bipyridyl, Copper (Ⅱ) chloride, Ammonium acetate, Pyrogallol, Quercetin, Folin & Ciocalteu's phenol reagent, Gallic acid, Vanillin, Catechin, TPTZ (2,4,6-tripyridyl-S-triazine ), Ferric Chloride was purchased and used from Sigma-Aldrich (St. Louis, Mo, USA), respectively. Glacial acetic acid was purchased and used from Alfa Aesar (Thermo Fisher Scientific), and sodium acetate trihydrate was purchased and used from JUNSEI.

세포 배양에 사용된 Dulbecco’s Modified Eagle’s Medium (DMEM), penicillin-streptomycin, phosphate buffered saline (PBS) 및 fetal bovine serum (FBS) 는 Hyclone 제품을 구입하였고 DMSO, L-Dopa 는 Sigma-Aldrich사 (St. Louis,Mo, USA) 에서 각각 구입하여 사용하였다. CCK-8 (Cell Counting Kit-8) 은 Dojindo(Molecular Technologies, Inc.) 에서 구입하여 사용하였다. 그 외 연구에 사용된 용매 및 시약은 일급 및 특급 시약을 구입하여 사용하였다.Dulbecco's Modified Eagle's Medium (DMEM), penicillin-streptomycin, phosphate buffered saline (PBS) and fetal bovine serum (FBS) used for cell culture were purchased from Hyclone, and DMSO and L-Dopa were purchased from Sigma-Aldrich (St. Louis). ,Mo, USA) respectively purchased and used. CCK-8 (Cell Counting Kit-8) was purchased and used from Dojindo (Molecular Technologies, Inc.). For other solvents and reagents used in the study, first-class and special-grade reagents were purchased and used.

2) Fe

Figure 112019049653517-pat00002
chelating ability2) Fe
Figure 112019049653517-pat00002
chelating ability

Fe chelating 측정은 Dinis et all.(1994) 의 방법을 변형하여 측정한다. 시료 120μl에 FeCl solution (30μl, 0.6mM), Bipyridyl solution (30μl, 5mM) 을 차례로 가하여 10분간 반응 후 562nm에서 흡광도를 측정하였다. Fe chelating measurement is measured by modifying the method of Dinis et all. (1994). FeCl solution (30μl, 0.6mM) and Bipyridyl solution (30μl, 5mM) were sequentially added to 120μl of the sample, reacted for 10 minutes, and absorbance was measured at 562nm.

3) Tyrosinase activity inhibition test3) Tyrosinase activity inhibition test

3차 증류수에 희석한 시료 40μl에 67mM Sodium phosphate buffer (pH6.8) 80μl를 가하여 완충시킨다. 만들어 놓은 buffer을 사용하여 10mM L-DOPA (3,4-dihydroxy-L-phenylalanine) 를 용해 시켜 40μl를 넣어준다. 마지막으로 125unit의 tyrosinase (from mushroom) 40μl를 가하여 37℃에서 10분간 반응 후 492nm에서 측정하였다. To 40 μl of the sample diluted in tertiary distilled water, 80 μl of 67 mM sodium phosphate buffer (pH6.8) was added to buffer. Using the prepared buffer, dissolve 10mM L-DOPA (3,4-dihydroxy-L-phenylalanine) and add 40μl. Finally, 40 μl of 125 units of tyrosinase (from mushroom) was added and reacted at 37° C. for 10 minutes, and then measured at 492 nm.

4) 세포배양4) Cell culture

실험에 사용한 세포는 Mouse melanoma melanocyte B16F0 세포를 ATCC에서 분양받아 사용하였다. B16F0 세포는 10% heat-inactivated fetal bovine serum (FBS), penicillin 및 2-mercaptoethanol이 함유된 Dulbecco’s Modified Eagle’s Medium(DMEM) 배지에서 5% CO2, 37℃조건에서 배양하여 실험에 사용하였다. As for the cells used in the experiment, Mouse melanoma melanocyte B16F0 cells were distributed from ATCC and used. B16F0 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) medium containing 10% heat-inactivated fetal bovine serum (FBS), penicillin and 2-mercaptoethanol at 5% CO 2 and 37°C, and used in the experiment.

5) 세포 생존율 측정5) Cell viability measurement

세포 생존율은 CCK-8 (Cell Counting Kit-8) 시약을 이용하여 측정하였다. 96-well plate에 B16F0 세포를 1 × 105 cells/well 농도로 접종한 후 각 well에 시료를 농도별로 투여하여 CO2 배양기에서 72시간 배양하였다. CCK-8 용액을 첨가하고 3시간 후 micro plate reader ((주)Bio-tech) 를 이용하여 450 nm에서 흡광도를 측정하여 대조그룹과 비교하였다. Cell viability was measured using CCK-8 (Cell Counting Kit-8) reagent. B16F0 cells were inoculated into a 96-well plate at a concentration of 1 × 10 5 cells/well, and then samples were administered to each well by concentration, followed by incubation in a CO 2 incubator for 72 hours. After 3 hours after adding the CCK-8 solution, the absorbance was measured at 450 nm using a micro plate reader (Bio-tech Co., Ltd.) and compared with the control group.

6) 세포내 tyrosinase 활성 저해 효과 측정6) Measurement of the inhibitory effect of intracellular tyrosinase activity

외부자극에 따른 B16F0세포 내 tyrosinase 의 활성에 실험시료가 영향을 미치는지를 알아보기 위해 외부자극제로 α-MSH (200 nM)을 사용하였으며 세포내 tyrosinase 활성 저해는 Matinez-Esparza의 방법을 변형시켜 측정하였다. 6-well plate에 1 × 106 cells/well로 세포를 분주하고 positive control로 α-MSH를 처리하여 자극하였다. 1시간 후 시료를 처리하였고, 72시간 배양한 후, 세포를 수집하여 1% (W/V) Triton X-100을 함유한 0.1 M sodium phosphate buffer (pH 6.5)를 1ml 가하여 용해하였다. 그 후 원심분리하여 얻은 상등액을 tyrosinase 활성과 단백질 정량에 이용하고, cell pellet은 멜라닌 정량에 사용하였다. 96-well plate에 시료 처리 후 얻은 상등액 100 μL를 분주하고 0.2 μg/ml 3,4-hydroxyphenylalamine (DOPA)가 첨가된 0.1 M sodium phosphate buffer (pH 6.5)를 100 μL씩 넣고 37℃에서 2시간 배양한 다음 490 nm에서 흡광도를 측정하였다.Α-MSH (200 nM) was used as an external stimulator to determine whether the test sample had an effect on the tyrosinase activity in B16F0 cells due to external stimulation, and the inhibition of intracellular tyrosinase activity was measured by modifying the method of Matinez-Esparza. . Cells were aliquoted into a 6-well plate at 1 × 10 6 cells/well and stimulated by treating α-MSH as a positive control. Samples were treated after 1 hour, and after 72 hours incubation, cells were collected and dissolved by adding 1 ml of 0.1 M sodium phosphate buffer (pH 6.5) containing 1% (W/V) Triton X-100. Then, the supernatant obtained by centrifugation was used for tyrosinase activity and protein quantification, and the cell pellet was used for melanin quantification. Dispense 100 μL of the supernatant obtained after sample treatment into a 96-well plate, add 100 μL of 0.1 M sodium phosphate buffer (pH 6.5) to which 0.2 μg/ml 3,4-hydroxyphenylalamine (DOPA) is added, and incubate at 37°C for 2 hours. Then, the absorbance was measured at 490 nm.

7) 멜라닌 정량7) Melanin quantification

Tyrosinase 활성을 측정하는 과정에서 얻은 pellet에 10% DMSO를 함유한 1N NaOH 용액 200 μL를 가하고 90℃에서 water bath에서 30분 방치하여 멜라닌을 완전히 용해시킨 후 475 nm에서 흡광도를 측정하였다. 멜라닌 양은 합성 멜라닌을 사용하여 작성된 표준 검량선에서 구하고, 실험그룹의 멜라닌 양은 대조그룹의 멜라닌 양에 대한 백분율로 계산하였다. 멜라닌 양은 각 well에서 측정한 단백질 농도를 기준으로 μg/mg protein으로 표기하였다.200 μL of a 1N NaOH solution containing 10% DMSO was added to the pellet obtained in the process of measuring the tyrosinase activity, and allowed to stand in a water bath at 90° C. for 30 minutes to completely dissolve melanin, and the absorbance was measured at 475 nm. The amount of melanin was obtained from a standard calibration curve prepared using synthetic melanin, and the amount of melanin in the experimental group was calculated as a percentage of the amount of melanin in the control group. The amount of melanin was expressed as μg/mg protein based on the protein concentration measured in each well.

8) 통계분석8) Statistical analysis

모든 결과는 3반복으로 실험하여 측정치를 평균값±표준편차로 나타내었으며 실험 결과의 통계적 유의성은 Student’s t-test로 하였으며 p 값이 0.05 미만일 때 통계적으로 유의성이 있다고 판단하였다. All the results were tested in three repetitions, and the measured values were expressed as the mean value ± standard deviation. The statistical significance of the experimental results was set to Student's t- test, and when the p value was less than 0.05, it was judged that there was statistical significance.

(p values *<0.05, **<0.01, ***<0.001 vs. control group, #<0.05, ##<0.01, ###<0.001 vs. α-MSH group $<0.05, $$<0.01, $$$<0.001 vs. (-)Glutathione group,)( p values * <0.05, ** <0.01, *** <0.001 vs. control group, # <0.05, ## <0.01, ### <0.001 vs. α-MSH group $ <0.05, $$ <0.01 , $$$ <0.001 vs. (-)Glutathione group,)

- 실험결과- Experiment result

1) Fe

Figure 112019049653517-pat00003
chelating ability1) Fe
Figure 112019049653517-pat00003
chelating ability

2가 철의 chelating 능력은 금속이온의 산화 반응을 유도하고 대조군과 실험군의 시료가 산화를 얼마나 저해 하는지를 확인하기 위한 실험이며, 그 결과 100배 희석농도에서 Glutathione 효모를 포함하지 않은 대조군(1.5%) 보다 Glutathione 효모를 포함한 실험군(1.5%)에서만 유의적인 저해효과를 나타내었다 (Fig. 4). The chelating ability of divalent iron is an experiment to induce an oxidation reaction of metal ions and to determine how much the samples of the control group and the experimental group inhibit oxidation. As a result, the control group that did not contain Glutathione yeast at a 100-fold dilution concentration (1.5%) More than that, only the experimental group (1.5%) containing Glutathione yeast showed a significant inhibitory effect (Fig. 4).

2) Tyrosinase inhibition rate2) Tyrosinase inhibition rate

세포내의 미백 활성을 확인하기 전 효소적 반응을 통한 이화학적 미백 활성 실험을 진행하였다. 체내의 미백관련 멜라닌 색소의 침착을 막기 위해서는 Tyrosinase의 활성을 낮춰야한다. 이러한 목적으로 Tyrosinase(from mushroom)을 이용하여 효소 활성 저해율을 측정하였다. 그 결과로 100배 희석액의 경우 Glutathione효모를 포함한 실험군(1.0%, 1.5%)에서 Glutathione 효모를 포함하지 않은 대조군에 비해 각각 1.0%에서는 16%, 1.5%에서는 12% 더 높은 저해율을 나타내었다 (Fig. 5).Before confirming the intracellular whitening activity, a physicochemical whitening activity experiment was conducted through an enzymatic reaction. In order to prevent the deposition of whitening-related melanin pigment in the body, it is necessary to lower the activity of Tyrosinase. For this purpose, the enzyme activity inhibition rate was measured using Tyrosinase (from mushroom). As a result, in the case of the 100-fold dilution, the experimental group (1.0%, 1.5%) containing Glutathione yeast showed a 16% higher inhibition rate than the control group not containing Glutathione yeast, respectively, 16% at 1.0% and 12% at 1.5% (Fig. 5).

3) B16F0세포의 생존율3) Survival rate of B16F0 cells

실험시료가 마우스 유래 악성 흑색종 세포인 B16F0 세포 생존율에 미치는 영향을 알아보기 위하여 X50배와 X100배의 희석배율을 선정하여 실험에 사용 하였으며 Figure 6의 결과, 세포 독성이 없음을 확인하였다 (Fig. 6). In order to investigate the effect of the experimental sample on the survival rate of mouse-derived malignant melanoma cells, B16F0 cells, X50 times and X100 times dilution ratios were selected and used in the experiment. As a result of Figure 6, it was confirmed that there was no cytotoxicity (Fig. 6).

4) 세포 내 tyrosinase 활성 저해 효과4) Inhibitory effect of intracellular tyrosinase activity

타이로시네이즈 (Tyrosinase) 는 멜라닌 합성 과정에서 속도결정단계인 초기 반응에 관여하는 가장 중요한 효소이다. 또한 멜라닌 합성의 주요한 단계를 나타내는 효소로 타이로신 (tyrosin) 을 DOPA로 전환하는 tyrosine hydroxylase 활성과 DOPA를 DOPAquinone으로 산화하는 DOPA oxidase 활성을 가지고 있다. 또한 매실추출물에서 tyrosinase 의 활성을 억제한다는 연구 결과가 보고 되어 있고 이에 대조군에 비해 글루타치온 효모를 함유한 실험시료의 경우 tyrosinase 활성을 얼마나 더 억제하는지 확인하고자 하였다. 또한 외부자극에 따른 B16FO세포 내tyrosinase의 활성 및 과생성멜라닌에 대해 실험시료의 영향을 확인하기 위해 외부자극제로 α-MSH (200 nM) 을 사용하였다. 그 결과 두 시료 모두 B16f0세포의 tyrosinase 의 활성에 크게 영향을 미치지 않는 것을 확인하였다 (Fig. 7). Tyrosinase is the most important enzyme involved in the initial reaction, which is the rate-determining step in the melanin synthesis process. In addition, it has tyrosine hydroxylase activity that converts tyrosine to DOPA as an enzyme representing a major step in melanin synthesis, and DOPA oxidase activity that oxidizes DOPA to DOPAquinone. In addition, studies have reported that plum extract inhibits the activity of tyrosinase, and this was to confirm how much more inhibited tyrosinase activity in the experimental sample containing glutathione yeast compared to the control group. In addition, α-MSH (200 nM) was used as an external stimulator to confirm the effect of experimental samples on tyrosinase activity and overproduced melanin in B16FO cells according to external stimulation. As a result, it was confirmed that both samples did not significantly affect the tyrosinase activity of B16f0 cells (Fig. 7).

5) 멜라닌 생합성 저해 효과5) Melanin biosynthesis inhibitory effect

B16F0 악성 흑색종 멜라노마 세포를 이용한 멜라닌 생합성 저해 효과는 Matinez-Esparza 의 방법을 변형시켜 측정하였다. B16F0 악성 흑색종 멜라노마 세포에 실험시료를 희석배율 별로 처리하고 72시간 배양한 다음 멜라닌 생성량을 측정하였다. 그 결과 대조군에 비해 글루타치온 효모를 함유한 실험시료의 경우 X50배로 희석한 그룹에서는 통계적으로 유의하게 멜라닌 생합성을 저해하는 효과를 나타내었지만 positive control그룹에 비교해서 통계적으로 유의하게 멜라닌 생합성을 저해하지 않았으며, X100 배로 희석하여 처리한 그룹에서는 크게 차이를 나타내지 않는 것을 확인하였다 (Fig. 8).The inhibitory effect of melanin biosynthesis using B16F0 malignant melanoma melanoma cells was measured by modifying the method of Matinez-Esparza. B16F0 malignant melanoma melanoma cells were treated with different dilution ratios and cultured for 72 hours, and then the amount of melanin production was measured. As a result, compared to the control group, the experimental sample containing glutathione yeast showed a statistically significant inhibitory effect on melanin biosynthesis in the X50-fold dilution group, but did not significantly inhibit melanin biosynthesis compared to the positive control group. , It was confirmed that there was no significant difference in the group treated by dilution by X100 times (Fig. 8).

6) 항산화 활성 측정6) Measurement of antioxidant activity

Glutathione 효모를 1.5% 함유한 대조군과 과립화를 위해 HPMC를 첨가한 실험군, 그리고 과립화한 후 H2O로 용해한 실험군의 항산화 활성을 비교하였다. 그 결과 대조군에 비해 HPMC를 첨가한 실험군, 과립화한 후 H2O로 용해한 실험군에서 유의한 항산화 활성의 차이는 없었다. 따라서 과립화로 인한 항산화 활성은 감소하지 않는 것을 확인하였다 (Fig. 9-12).The antioxidant activity of the control group containing 1.5% glutathione yeast, the experimental group to which HPMC was added for granulation, and the experimental group dissolved in H 2 O after granulation were compared. As a result, there was no significant difference in antioxidant activity in the experimental group to which HPMC was added and the experimental group to which H 2 O was dissolved after granulation compared to the control group. Therefore, it was confirmed that the antioxidant activity did not decrease due to granulation (Fig. 9-12).

7) Tyrosinase inhibition rate7) Tyrosinase inhibition rate

Glutathione 효모를 1.5% 함유한 대조군과 과립화를 위해 HPMC를 첨가한 실험군, 그리고 과립화한 후 H2O로 용해한 실험군의 Tyrosinase (from mushroom) 을 이용하여 효소 활성 저해율을 측정하였다. 그 결과 100배 희석한 그룹에서는 대조군에 비해 HPMC를 첨가한 실험군, 과립화 한 후 H2O로 용해한 실험군에서 유의한 항산화 활성의 차이는 없었다. 따라서 과립화로 인한 항산화 활성은 감소하지 않는 것을 확인하였다.The inhibition rate of enzyme activity was measured using Tyrosinase (from mushroom) of the control group containing 1.5% glutathione yeast, the experimental group to which HPMC was added for granulation, and the experimental group dissolved in H 2 O after granulation. As a result, in the 100-fold diluted group, there was no significant difference in antioxidant activity in the experimental group to which HPMC was added and the experimental group to which H 2 O was dissolved after granulation compared to the control group. Therefore, it was confirmed that the antioxidant activity did not decrease due to granulation.

본 발명의 항산화 및 미백 조성물Antioxidant and whitening composition of the present invention

본 발명은 매실과 글루타치온(Glutathione) 효모의 혼합물을 과립화하며 HPMC를 첨가한다.In the present invention, a mixture of plum and glutathione yeast is granulated and HPMC is added.

상기 HPMC는 propylene glycol ether of methylcellulose 의 약자로, 화학적 구조는 셀룰로오스 주사슬로 구성되어 있고, 여기서 치환체 R은 H, -CH3이거나 -CH2CH(CH3)OH 이다.The HPMC stands for propylene glycol ether of methylcellulose, and its chemical structure is composed of a cellulose main chain, where the substituent R is H, -CH3 or -CH2CH(CH3)OH.

Figure 112019049653517-pat00004
Figure 112019049653517-pat00004

셀룰로오스는 하이드록시 그룹의 강한 수소결합으로 인해 결정을 형성하여 물에 잘 녹지 않지만 HPMC는 비이온성을 나타내어 물에 대한 용해도가 높고 낮은 온도에서도 잘 용해되어, 점성이 있는 콜로이드 용액을 형성한다. 클로로포름, 에탄올, 에테르 등과 같은 유기 용매에는 거의 용해되지 않으나 에탄올과 디클로로메탄, 메탄올과 디클로로메탄, 물과 알콜의 혼합액에는 용해된다. 그리고 일반적으로 무미, 무취의 백색 분말로 자극성, 독성이 없으며 인체에 무해하다.Cellulose forms crystals due to strong hydrogen bonds of hydroxy groups and is not well soluble in water, but HPMC has high solubility in water and is well soluble at low temperatures, forming a viscous colloidal solution. It is hardly soluble in organic solvents such as chloroform, ethanol, and ether, but it is soluble in a mixture of ethanol and dichloromethane, methanol and dichloromethane, and water and alcohol. In general, it is a tasteless, odorless white powder that is not irritating or toxic, and is harmless to the human body.

Figure 112019049653517-pat00005
Figure 112019049653517-pat00005

HPMC의 물리화학적 특성은 i) -OCH3 그룹의 비, ii) -OCH2CH(CH3)OH 그룹의 비, iii) 분자량과 관계가 있다. 미국약전(USP)에서는 HPMC의 종류를 표 1과 같이 -OCH3, -OCH2CH(CH3)OH의 비율에 따라서 HPMC1828, HPMC2208(K), HPMC2906(F), HPMC2910(E)으로 정의하고 있다. 앞의 두 숫자는 -OCH3 그룹의 비율이고, 뒤의 두 숫자는 -OCH2CH(CH3)OH 그룹의 비율이다. 모든 등급의 HPMC가 다양한 제제에서 널리 이용되는데, 특히 약물방출제어에 사용되는 것은 주로 상대적으로 점도가 높은 2208과 2910등급이다.The physicochemical properties of HPMC are related to i) the ratio of -OCH3 groups, ii) the ratio of -OCH2CH(CH3)OH groups, and iii) the molecular weight. The United States Pharmacopoeia (USP) defines the types of HPMC as HPMC1828, HPMC2208(K), HPMC2906(F), and HPMC2910(E) according to the ratio of -OCH3, -OCH2CH(CH3)OH as shown in Table 1. The first two numbers are the ratio of the -OCH3 group, and the second two numbers are the ratio of the -OCH2CH(CH3)OH group. All grades of HPMC are widely used in various formulations, especially those used for drug release control are mainly grades 2208 and 2910, which have relatively high viscosity.

미국약전(USP)에서는 HPMC의 점도측정법을 20 ℃에서 2%용액으로 만들어 측정한다고 제시하고 있다. 이때 HPMC의 점도는 측정온도에 따라 크게 변하게 되는데 그 이유는 HPMC가 특정 온도에서 젤로 변화하는 성질을 가지고 있기 때문이다. 온도에 따른 졸-젤-졸로의 상전이 현상은 분자내의 소수성 작용기들 간의 상호작용에 의해 중합체가 형성되기 때문이다. 또 다른 분류 방법으로는 HPMC의 등급에 따라 P: premium, LV: low viscosity, CR: controlled release, G: granular, S: surface treated, FG: food grade 등으로 나눠지기도 한다.The US Pharmacopoeia (USP) suggests that HPMC's viscosity measurement method is made into a 2% solution at 20°C. At this time, the viscosity of HPMC varies greatly depending on the measurement temperature, because HPMC has a property of changing to a gel at a specific temperature. The sol-gel-zolo phase transition phenomenon according to temperature is because a polymer is formed by the interaction between hydrophobic functional groups in the molecule. Another classification method is classified into P: premium, LV: low viscosity, CR: controlled release, G: granular, S: surface treated, and FG: food grade, depending on the grade of HPMC.

본 발명에서 HPMC는 매실과 글루타치온 효모를 코팅하며 결합하여 과립화하는 결합제로 사용하였다. 이러한 구성에 의하여 글루타치온 효모에서 발생하는 이취를 섭취 시 복용자가 느낄 수 없도록 하였다. In the present invention, HPMC was used as a binder to coat, combine, and granulate plum and glutathione yeast. This configuration prevented the user from feeling the off-flavor generated from glutathione yeast when ingested.

본 발명은 사용자의 섭취 편의와 기능을 향상하기 위하여 매실농축액, 결정과당, 자일리톨, 미네랄, 비타민, 글루타치온 효모(글루타치온 50% 함유), 포도당을 하기 샘플1 내지 3의 비율로 혼합하여 사용하였다.In the present invention, plum concentrate, crystal fructose, xylitol, minerals, vitamins, glutathione yeast (containing 50% glutathione), and glucose were mixed and used in the following samples 1 to 3 in order to improve the convenience and function of the user's intake.

본 발명의 항산화 및 미백 조성물은 매실 5 중량%, 자일리톨 50 중량%, 결정과당 29.5 중량%, 비타민C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 0.5 중량% 인 것을 특징으로 하는 항산화 및 미백 조성물을 제공한다.The antioxidant and whitening composition of the present invention is 0.5% by weight of glutathione yeast containing 5% by weight of plum, 50% by weight of xylitol, 29.5% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 0.5% by weight of glutathione. It provides an antioxidant and whitening composition.

또 다른 실시예로, 본 발명의 항산화 및 미백 조성물은 매실 5 중량%, 자일리톨 50 중량%, 결정과당 29 중량%, 비타민C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 1 중량% 인 것을 특징으로 하는 항산화 및 미백 조성물을 제공한다.In another embodiment, the antioxidant and whitening composition of the present invention is a glutathione yeast containing 5% by weight of plum, 50% by weight of xylitol, 29% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 50% by weight of glutathione. It provides an antioxidant and whitening composition, characterized in that 1% by weight.

또 다른 실시예로, 본 발명의 항산화 및 미백 조성물은 매실 5 중량%, 자일리톨 50 중량%, 결정과당 28.5 중량%, 비타민C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 1.5 중량% 인 것을 특징으로 하는 항산화 및 미백 조성물을 제공한다.In another embodiment, the antioxidant and whitening composition of the present invention is a glutathione yeast containing 5% by weight of plum, 50% by weight of xylitol, 28.5% by weight of fructose, 7% of vitamin C, 3% by weight of minerals, and 50% by weight of glutathione. It provides an antioxidant and whitening composition, characterized in that 1.5% by weight.

Figure 112021013209169-pat00017
Figure 112021013209169-pat00017

상기 배합비로 배합된 조성물을 하기의 방법으로 과립화하였다.The composition blended in the above blending ratio was granulated by the following method.

매실농축액 (10 Bix)를 기반으로 자일리톨, 결정과당, 비타민C, 미네랄 글루타치온 효모 분말(글루타치온 50% 함유)를 배합하여 액상 재료를 준비하는 혼합조성물 제조단계(S1); 및 A mixed composition preparation step (S1) of preparing a liquid material by mixing xylitol, crystal fructose, vitamin C, and mineral glutathione yeast powder (containing 50% glutathione) based on plum concentrate (10 Bix); And

유동층 과립기의 공정온도를 50℃로 설정하여 슈가 시드(sugger seed)을 과립화를 위한 핵으로 사용하여 과립화하는 과립화공정(S2); A granulation step (S2) of granulating using a sugar seed as a nucleus for granulation by setting the process temperature of the fluid bed granulator to 50°C;

상기 과립화공정에서 과립화된 조성물 과립을 HMPC로 코팅하여 상기 과립이 수분을 흡수하여 뭉치는 것을 방지하는 HMPC코팅공정(S3);을 포함하는 것을 특징으로 하는 매실과 글루타치온 효모를 구비한 항산화 및 미백 조성물을 제공한다. Antioxidant comprising plum and glutathione yeast, comprising: an HMPC coating process (S3) in which the granules of the composition granulated in the granulation process are coated with HMPC to absorb moisture and prevent agglomeration of the granules. Provides a whitening composition.

상기 슈가 시드의 직경은 0.3mm의 것을 사용하는 것을 특징으로 하는 매실과 글루타치온 효모를 구비한 항산화 및 미백 조성물을 제공한다.It provides an antioxidant and whitening composition comprising plum and glutathione yeast, characterized in that the sugar seed has a diameter of 0.3mm.

상기 공정온도는 60℃ 이하인 것을 특징으로 한다. 60℃가 되면 상기 혼합조성물이 모두 용해되어 과립화가 불가능하기 때문이다.The process temperature is characterized in that less than 60 ℃. This is because when the temperature reaches 60° C., all of the mixed composition is dissolved and granulation is impossible.

삭제delete

Claims (3)

매실 5 중량%, 자일리톨 50 중량%, 결정과당 29.5 중량%, 비타민C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 0.5 중량%를 배합하여 액상 재료를 준비하는 혼합조성물 제조단계(S1); 및
유동층 과립기의 공정온도를 50℃로 설정하여 슈가 시드(sugger seed)을 과립화를 위한 핵으로 사용하여 과립화하는 과립화공정(S2);
상기 과립화공정에서 과립화된 조성물 과립을 HMPC로 코팅하여 상기 과립이 수분을 흡수하여 뭉치는 것을 방지하는 HMPC코팅공정(S3);을 포함하는 것을 특징으로 하는 매실과 글루타치온 효모를 구비한 항산화 및 미백 조성물.
A mixed composition prepared by mixing 5% by weight of plum, 50% by weight of xylitol, 29.5% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 0.5% by weight of glutathione yeast containing 50% by weight of glutathione Step (S1); And
A granulation step (S2) of granulating using a sugar seed as a nucleus for granulation by setting the process temperature of the fluid bed granulator to 50°C;
Antioxidant comprising plum and glutathione yeast, comprising: an HMPC coating process (S3) in which the granules of the composition granulated in the granulation process are coated with HMPC to absorb moisture and prevent agglomeration of the granules. Whitening composition.
매실 5 중량%, 자일리톨 50 중량%, 결정과당 29 중량%, 비타민C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 1 중량% 를 배합하여 액상 재료를 준비하는 혼합조성물 제조단계(S1); 및
유동층 과립기의 공정온도를 50℃로 설정하여 슈가 시드(sugger seed)을 과립화를 위한 핵으로 사용하여 과립화하는 과립화공정(S2);
상기 과립화공정에서 과립화된 조성물 과립을 HMPC로 코팅하여 상기 과립이 수분을 흡수하여 뭉치는 것을 방지하는 HMPC코팅공정(S3);을 포함하는 것을 특징으로 하는 매실과 글루타치온 효모를 구비한 항산화 및 미백 조성물.
A mixed composition prepared by mixing 5% by weight of plum, 50% by weight of xylitol, 29% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 1% by weight of glutathione yeast containing 50% by weight of glutathione Step (S1); And
A granulation step (S2) of granulating using a sugar seed as a nucleus for granulation by setting the process temperature of the fluid bed granulator to 50°C;
Antioxidant comprising plum and glutathione yeast, comprising: an HMPC coating process (S3) in which the granules of the composition granulated in the granulation process are coated with HMPC to absorb moisture and prevent agglomeration of the granules. Whitening composition.
매실 5 중량%, 자일리톨 50 중량%, 결정과당 28.5 중량%, 비타민C 7%, 미네랄 3 중량%, 50 중량%의 글루타치온을 함유하는 글루타치온 효모 1.5 중량% 를 배합하여 액상 재료를 준비하는 혼합조성물 제조단계(S1); 및
유동층 과립기의 공정온도를 50℃로 설정하여 슈가 시드(sugger seed)을 과립화를 위한 핵으로 사용하여 과립화하는 과립화공정(S2);
상기 과립화공정에서 과립화된 조성물 과립을 HMPC로 코팅하여 상기 과립이 수분을 흡수하여 뭉치는 것을 방지하는 HMPC코팅공정(S3);을 포함하는 것을 특징으로 하는 매실과 글루타치온 효모를 구비한 항산화 및 미백 조성물.
A mixed composition prepared by mixing 5% by weight of plum, 50% by weight of xylitol, 28.5% by weight of crystalline fructose, 7% of vitamin C, 3% by weight of minerals, and 1.5% by weight of glutathione yeast containing 50% by weight of glutathione Step (S1); And
A granulation step (S2) of granulating using a sugar seed as a nucleus for granulation by setting the process temperature of the fluid bed granulator to 50°C;
Antioxidant comprising plum and glutathione yeast, comprising: an HMPC coating process (S3) in which the granules of the composition granulated in the granulation process are coated with HMPC to absorb moisture and prevent agglomeration of the granules. Whitening composition.
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