KR20150072567A - Method for estimating skin whitening effect of cosmetic composition - Google Patents

Method for estimating skin whitening effect of cosmetic composition Download PDF

Info

Publication number
KR20150072567A
KR20150072567A KR1020130159835A KR20130159835A KR20150072567A KR 20150072567 A KR20150072567 A KR 20150072567A KR 1020130159835 A KR1020130159835 A KR 1020130159835A KR 20130159835 A KR20130159835 A KR 20130159835A KR 20150072567 A KR20150072567 A KR 20150072567A
Authority
KR
South Korea
Prior art keywords
cosmetic composition
melanin
skin
amount
stratum corneum
Prior art date
Application number
KR1020130159835A
Other languages
Korean (ko)
Other versions
KR102134930B1 (en
Inventor
강혜리
박소현
김성우
조진훈
Original Assignee
코웨이 주식회사
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 코웨이 주식회사 filed Critical 코웨이 주식회사
Priority to KR1020130159835A priority Critical patent/KR102134930B1/en
Publication of KR20150072567A publication Critical patent/KR20150072567A/en
Application granted granted Critical
Publication of KR102134930B1 publication Critical patent/KR102134930B1/en

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pathology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Food Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Dermatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Toxicology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Cosmetics (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The present invention relates to a method for estimating a skin whitening effect of cosmetic compositions by measuring an amount of melanine on keratin layers after coating cosmetic composition on the kerain layers wherein the method can objectively evaluate skin whitening effect of cosmetic compositions while reducing time of measuring the amount and long-process of stimulating skin of customers.

Description

화장료 조성물의 피부 미백 효과 평가 방법{Method for estimating skin whitening effect of cosmetic composition}[0001] The present invention relates to a method for evaluating skin whitening effect of a cosmetic composition,

본 발명은 피부에 화장료 조성물을 도포한 후 각질층에 존재하는 멜라닌 양을 측정함으로써 화장료 조성물의 피부 미백 효과를 평가하는 방법이다. The present invention is a method for evaluating the skin whitening effect of a cosmetic composition by measuring the amount of melanin present in the stratum corneum after applying the cosmetic composition to the skin.

멜라닌은 피부, 털, 눈 등에 존재하는 흑갈색 알갱이 색소로서 멜라닌의 양에 따라 피부색이 결정된다. 또한 멜라닌은 일정량 이상의 자외선을 흡수하여 유해한 자외선이 인체 내로 침투하는 것을 차단하여 인체를 보호하는 역할을 한다. Melanin is a blackish brown granular pigment present in skin, hair, eyes, etc., and its skin color is determined by the amount of melanin. In addition, melanin absorbs more than a certain amount of ultraviolet rays, and protects the human body from harmful ultraviolet rays penetrating into the human body.

피부의 진피와 표피의 경계선 역할을 하는 기저막 위에 위치하는 멜라닌 형성 세포는 진피층에 위치한 섬유아세포의 영향을 받아 멜라닌을 생성한다. 멜라닌 형성 세포에서 생성된 멜라닌은 세포 돌기를 통해 주변의 각질 형성세포로 이동하여 피부에 넓게 분산됨으로써 자외선과 같은 유해 인자로부터 피부를 보호하고 피부색을 나타낸다. 정상 피부는 자외선과 같은 자극이 왔을 때 멜라닌 형성 세포가 평소보다 많은 멜라닌을 생성하고 분산하여 자외선이 피부 깊숙이 침투하는 것을 방지한다. 자극이 소멸하는 경우 멜라닌의 생성과 분산은 다시 정상 수준으로 돌아온다.
Melanocytes located on the basement membrane, which act as a boundary between the dermis and epidermis of the skin, are affected by the fibroblasts located in the dermis and produce melanin. Melanin generated from melanin-forming cells migrates to surrounding keratinocytes through cell protrusions and is widely dispersed in the skin, thereby protecting skin from harmful factors such as ultraviolet rays and exhibiting skin color. Normal skin, when stimuli such as ultraviolet light, melanin-forming cells produce and disperse more melanin than usual to prevent penetration of ultraviolet rays deep into the skin. When the stimulus disappears, melanin production and dispersion returns to normal levels.

한국인과 일본인과 같은 동양인들은 맑고 깨끗한 피부에 대한 관심이 높다. 이런 분위기를 반영하듯이 국내화장품업계뿐만 아니라 다국적 기업도 미백 원료 물질이나 효과에 연구가 증가하고 있다. Oriental people like Koreans and Japanese are interested in clear and clean skin. As reflected in this atmosphere, not only the domestic cosmetics industry but also the multinational companies are increasingly studying the whitening raw materials and effects.

미백 원료 물질로는 멜라닌 색소 생성에 중요한 작용을 하는 효소인 티로시나제를 억제하거나 항산화 작용을 가지는 원료들이 주로 사용되고 있다. Raw materials that inhibit tyrosinase, an enzyme that plays an important role in the production of melanin pigment, or antioxidant substances are mainly used as a whitening raw material.

이에 화장료 조성물의 미백 효과를 평가하기 위해 티로시나제 활성억제를 평가하는 방법이 많이 사용되고 있다. 하지만 단지 시험관 상에서 티로시나제 활성 억제 능력을 가진 물질들이 멜라닌 세포를 대상으로 한 실험에서 더 나아가 임상에서 같은 정도의 효과를 나타낼지는 장담하기 어렵다. 또한 최근 티로시나제를 직접 억제하지 않더라도 우수한 미백효과를 나타내는 물질들이 많이 연구되고 있다.Therefore, a method of evaluating inhibition of tyrosinase activity in order to evaluate the whitening effect of a cosmetic composition is widely used. However, it is difficult to say whether substances that have the ability to inhibit tyrosinase activity in vitro are further tested in melanocyte cells and show the same effect in clinical practice. Recently, many substances that exhibit excellent whitening effect even without directly inhibiting tyrosinase have been studied.

따라서 티로시나제 억제효과만을 미백 기능성 화장품의 유효성 평가방법으로 사용하는 데에는 무리가 있으며, 새로운 평가기준의 개발이 시급한 실정이다. Therefore, it is difficult to use only the inhibitory effect of tyrosinase as a method for evaluating the effectiveness of whitening functional cosmetics, and it is urgent to develop a new evaluation standard.

미백 화장품의 미백 효능을 평가하는 임상 평가 방법으로 다음과 같은 방법이 알려져 있다. The following methods are known as clinical evaluation methods for evaluating whitening efficacy of whitening cosmetics.

피험자의 피부에 인공적으로 자외선을 조사하여 색소 침착을 야기한 후에 시료를 도포하여 그 효능 여부를 결정하는 평가방법이 있다. 하지만, 자외선 조사에 의하여 피험자의 피부 표면에 수포가 발생할 정도의 과도한 홍반 반응이 발생할 수 있으며, 광알레르기 또는 광감작의 나타날 가능성도 배제할 수 없기 때문에, 시험에 참여하는 피험자에게는 부담이 되는 것이 사실이다. There is an evaluation method of irradiating ultraviolet rays to the skin of the subject artificially to cause pigmentation and then applying the sample to determine the efficacy. However, since the ultraviolet irradiation may cause an excessive erythema reaction that causes blisters on the skin surface of the subject, and it is impossible to exclude the possibility of optical allergy or light sensitization, it is a burden to the subjects participating in the test to be.

화장품의 미백 효능 평가는 기미, 점, 흑자, 검버섯과 같은 과색소침착증상을 가진 사람을 대상으로 이루어질 수도 있다. 하지만, 이러한 피부 과색소침착증은 그 발생 요인이나 기전이 자외선에 의한 멜라닌 생성 증가와 다르며, 비록 자외선에 의해 그 증상이 악화되는 경향은 있으나 자외선과 같은 자극이 없는 경우에도 멜라닌 생성과 분산이 계속적으로 증가한다. The evaluation of whitening efficacy of cosmetics may be made for people with hyperpigmentation symptoms such as spots, points, surplus, and black spots. However, this skin hypercholesterolemia is different from the increase of the production of melanin by ultraviolet rays, and the cause and mechanism of the hypercholesterolemia are different. Although the symptoms tend to be worsened by ultraviolet rays, melanin generation and dispersion continuously occur even when there is no stimulation such as ultraviolet light .

또한 두 가지의 평가 방법은 피험자의 비교적 긴 기간 동안의 자율적인 화장품 도포를 전제로 하고 있으므로, 화장품의 도포가 평가자의 기준에 따라 적합하게 이루어지고 있는지 확인하기 힘들다. In addition, since the two methods of evaluation are based on autonomous application of cosmetics over a relatively long period of time, it is difficult to confirm whether the application of cosmetics is appropriately performed according to the evaluator's standard.

이에, 시험의 부작용에 대한 피험자의 부담이 적으며, 화장품의 도포가 평가자에 의해 일관되게 이루어지고, 평가 기간이 짧으면서도 객관적인 결과를 도출해낼 수 있는 피부 미백 화장료 조성물에 대한 임상 평가 방법의 요구가 존재한다. Therefore, there is a demand for a clinical evaluation method for a skin whitening cosmetic composition which is less burdensome for the adverse reaction of the test, the application of the cosmetic is consistently performed by the evaluator, and an objective result can be obtained with a short evaluation period exist.

본 발명의 주된 과제는 피험자에게 피부에 자극을 주는 과정과 긴 시험 기간의 부담을 덜 수 있을 뿐만 아니라 명확하고 객관적으로 화장료 조성물의 피부 미백 효과를 평가할 수 있는 평가 방법을 제공하는 것이다. The main object of the present invention is to provide a method for evaluating a skin whitening effect of a cosmetic composition clearly and objectively as well as a process of stimulating skin to a subject and a burden of a long test period.

이를 위해 본 발명은To this end,

화장료 조성물의 피부 미백 효과를 평가하는 방법으로서,A method for evaluating the skin whitening effect of a cosmetic composition,

화장료 조성물 도포 전과 도포 후 피부 각질층의 멜라닌 양을 측정함으로써 화장료 조성물의 피부 미백 효과를 평가하는 방법을 제공한다.The present invention provides a method for evaluating the skin whitening effect of a cosmetic composition by measuring the amount of melanin in the stratum corneum before and after application of the cosmetic composition.

본 발명의 평가 방법에 의하면, 화장료 조성물이 누적 첩포 방법으로 도포되므로, 기존에 이루어지던 임상 평가에 비해 평가 기간이 단축되고, 평가자에 의한 일관된 도포가 이루어지기 때문에 피험자의 화장품 사용 습관이나 빈도 등에 따른 결과의 차이가 감소된다. 또한, 테이프 스트리핑 방법으로 평가 시료를 채취하기 때문에, 피험자의 부담이 적고, 누구든지 간단하게 평가하는 것이 가능해진다. 또한 각질층의 멜라닌 양을 측정하기 때문에 평가의 신뢰성을 증가시킬 수 있다. According to the evaluation method of the present invention, since the cosmetic composition is applied by the cumulative deposition method, the evaluation period is shortened compared with the conventional clinical evaluation, and the uniform application by the evaluator is performed. The difference in results is reduced. Further, since the evaluation sample is sampled by the tape stripping method, the burden on the subject is small, and anyone can easily evaluate it. Also, since the amount of melanin in the stratum corneum is measured, the reliability of the evaluation can be increased.

도 1은 평가시험 전 비교제형 및 실험제형 도포 부위에서 채취한 각질층에 존재하는 멜라닌 양을 나타낸 도면이다.
도 2는 누적 첩포에 의한 비교제형 및 실험제형 도포 후 도포 부위에서 채취한 각질층에 존재하는 멜라닌 양을 나타낸 도면이다.
도 3은 비교제형 및 실험제형의 멜라닌 감소율을 비교하여 나타낸 도면이다. FIG. 1 is a graph showing the amount of melanin present in the stratum corneum collected from the comparative formulation before the evaluation test and the test formulation application site.
FIG. 2 is a graph showing the amount of melanin present in the stratum corneum collected at the application site after application of the comparative formulation and experimental formulation by the cumulative pellet.
FIG. 3 is a graph comparing melanin reduction rates of the comparative formulation and the experimental formulation.

이하 본 발명을 더욱 상세히 설명한다. Hereinafter, the present invention will be described in more detail.

본 발명의 평가 방법은, 화장료 조성물의 피부 미백 효과를 평가하기 위해 피부 각질층의 멜라닌 양의 많고 적음을 지표로 하는 것을 특징으로 하고 있다. The evaluation method of the present invention is characterized in that the amount of melanin in the skin stratum corneum is increased or decreased in order to evaluate the skin whitening effect of the cosmetic composition.

멜라닌 생성이 억제된 피부는 그렇지 않은 피부에 비하여 멜라닌의 양이 유의하게 낮기 때문에, 각질층에 존재하는 멜라닌 양의 많고 적음을 지표로 하여, 간편하게 화장품의 미백 효능을 평가 할 수 있다.Since the amount of melanin in the skin inhibited by melanin production is significantly lower than that of the skin in which the melanin production is inhibited, the whitening effect of the cosmetic product can be easily evaluated using the amount of melanin present in the stratum corneum as an index.

이를 위해 화장료 조성물 도포 전과 도포 후 피부 각질층의 멜라닌 양을 측정한다. For this purpose, the amount of melanin in the stratum corneum of the skin before and after application of the cosmetic composition is measured.

각질층은 피부의 가장 위에 있는 조직으로, 체외로부터의 이물질이나 자극으로부터 피부를 지키는 작용을 가지고 있다. The stratum corneum is the uppermost tissue in the skin and has the function of protecting the skin from foreign substances or stimuli.

본 발명에 있어서의 평가 대상 부위는, 각질층을 입수할 수 있는 부분이라면 어떠한 부위도 포함될 수 있지만, 주된 부위로서는 전완 또는 등 부위를 들 수 있다.
The evaluation target site in the present invention may include any site as long as it can acquire the stratum corneum, but the main site may be the forearm or back region.

본 발명에서 사용하는 화장료 조성물 도포 방법은, 누적 첩포 방법(Repeated Insult Patch Test)을 이용한다. 피험자의 전완 또는 등 부위에 48시간 첩포를 주 2회, 4주간 총 8회 적용하며, 화장료 도포 전과 도포 후 각질층을 채취한다. The method for applying the cosmetic composition used in the present invention uses a Repeated Insult Patch Test. Apply 48 hours of papillon twice a week or 8 times for 4 weeks to the subject's forearm or back, and collect the stratum corneum before and after application of cosmetics.

각질층 채취는 테이프로 각질층의 표층 부분만을 채취하는 테이프 스트리핑 방법으로 간단하게 수행 할 수 있다. 이 때문에 사용자에게 부담을 주지 않고, 사용자의 피부에 대한 화장품의 미백 효능 여부를 평가할 수 있다. The stratum corneum collection can be simply performed by a tape stripping method in which only the surface layer portion of the stratum corneum is extracted with a tape. Therefore, it is possible to evaluate whether or not the whitening effect of the cosmetic product on the user's skin can be evaluated without imposing a burden on the user.

즉, 화장료 조성물이 도포된 부위에 스트리핑 테이프를 일정 시간, 즉 피부 각질층 채취에 충분한 시간 동안, 예를 들어 10초 내지 7분 동안 부착했다가 떼어냄으로써 피부 각질층을 채취할 수 있다. That is, the stratum corneum can be collected by attaching the stripping tape to the site to which the cosmetic composition is applied for a predetermined time, that is, for a sufficient time for taking a horny layer of the skin, for example, for 10 seconds to 7 minutes and then removing it.

이렇게 채취된 피부 각질층으로부터 멜라닌을 추출한다. 생화학적 방법, 예를 들면 동결 융해법, 초음파 파쇄법, 호모제네이트법(homogenate method)등을 매개로 하여 각 시료로부터 멜라닌을 추출한다. The melanin is extracted from the skin stratum corneum thus taken. Melanin is extracted from each sample through biochemical methods such as freeze-thaw method, ultrasonic disruption method, and homogenate method.

멜라닌의 양은, 종래부터 알려져 있는 방법으로 측정할 수 있다. 예를 들면, 멜라닌에 대한 항체와의 반응에 기초하는 효소 결합 면역 분석법(ELISA, Enzyme-linked immunosorbent assay), 방사면역분석법(radioimmunoassay), 웨스턴 블로팅(western blotting) 등의 방법을 이용 할 수 있다.
The amount of melanin can be measured by a conventionally known method. For example, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, Western blotting and the like based on the reaction with an antibody against melanin can be used .

본 발명의 평가 방법은 구체적으로,The evaluation method of the present invention is, specifically,

(S1) 피험자 각질층을 채취하여 각질층의 멜라닌 양을 측정하는 단계;(S1) measuring the amount of melanin in the stratum corneum by taking a stratum corneum of the subject;

(S2) 피부에 시험 대상 화장료 조성물을 도포한 후 피부 각질층을 채취하여 각질층의 멜라닌 양을 측정하는 단계; 및(S2) measuring the amount of melanin in the stratum corneum after applying the test cosmetic composition to the skin and collecting the stratum corneum of the skin; And

(S3) 상기 1) 단계 및 2) 단계에서 측정한 멜라닌양의 함량변화로부터 도포된 화장료 조성물의 피부 미백 효과를 평가하는 단계를 거친다.
(S3) A step of evaluating the skin whitening effect of the applied cosmetic composition from the change of the content of melanin amount measured in steps 1) and 2) is performed.

이때 시험 대상 화장료 조성물은 비교제형 화장료 조성물과 평가하고자 하는 실험제형 화장료 조성물을 사용할 수 있다. In this case, the cosmetic composition to be tested may be a comparative cosmetic composition and an experimental cosmetic composition to be evaluated.

각 화장료 조성물 도포 전후 측정된 멜라닌 감소율을 비교함으로써 실험제형 화장료 조성물의 피부 미백 효과를 객관적으로 평가할 수 있다.The skin whitening effect of the experimental cosmetic composition can be objectively evaluated by comparing the measured decrease rate of melanin before and after application of each cosmetic composition.

즉, 실험제형 화장료 조성물을 도포한 부위에서 측정된 멜라닌 감소율이 비교제형 화장료 조성물을 도포한 부위에서 측정된 멜라닌 감소율 보다 유의하게 높은 경우, 피부 미백 효능이 나타났다고 평가 할 수 있다.
That is, when the melanin reduction rate measured at the site where the experimental cosmetic composition is applied is significantly higher than the melanin reduction rate measured at the site where the comparative cosmetic composition is applied, skin whitening efficacy can be evaluated.

본 발명의 평가 방법에 의하면, 화장료 조성물이 누적 첩포 방법으로 도포되므로, 기존에 이루어지던 임상 평가에 비해 평가 기간이 단축되고, 평가자에 의한 일관된 도포가 이루어지기 때문에 피험자의 화장품 사용 습관이나 빈도 등에 따른 결과의 차이가 감소된다. 또한, 테이프 스트리핑 방법으로 평가 시료를 채취하기 때문에, 피험자의 부담이 적고, 누구든지 간단하게 평가하는 것이 가능해진다. 또한 각질층의 멜라닌 양을 측정하기 때문에 평가의 신뢰성을 증가시킬 수 있다.
According to the evaluation method of the present invention, since the cosmetic composition is applied by the cumulative deposition method, the evaluation period is shortened compared with the clinical evaluation that has been performed previously, and the uniform application by the evaluator is performed. The difference in results is reduced. Further, since the evaluation sample is sampled by the tape stripping method, the burden on the subject is small, and anyone can easily evaluate it. Also, since the amount of melanin in the stratum corneum is measured, the reliability of the evaluation can be increased.

바람직한 구현예로서, 본 발명에서 제시한 방법에 따라 화장품에 대한 피부 미백 효능을 분석하였다.
As a preferred embodiment, skin whitening efficacy on cosmetics was analyzed according to the method presented in the present invention.

[비교제형 및 실험제형 제조] [Preparation of Comparative Formulations and Experimental Formulations]

본 실험을 위하여 준비된 화장료 제형의 제법은 다음과 같다.The preparation of cosmetic formulation prepared for this experiment is as follows.

아래 표 1의 성분에 따라 수상, 유상을 75 ℃까지 가온하여 완전 용해한 후 아지 믹서(Agi Mixer)를 이용하여 수상을 스터링하면서 유상을 투입하였다. 유상 투입 후 5분간 3000rpm으로 아지 믹서로 혼합하였다. 혼합 후 중화 상을 투입하여 5분간 3000rpm으로 아지 믹서로 혼합하였다. 혼합 후 30 ℃까지 냉각하였다.According to the ingredients shown in Table 1 below, the water phase and the oil phase were completely dissolved by heating to 75 DEG C, and the aqueous phase was added while stirring the aqueous phase using an Agi Mixer. After the oil phase injection, they were mixed with an agitator at 3000 rpm for 5 minutes. After mixing, the neutralization phase was added and mixed for 5 minutes at 3000 rpm with an agitator. The mixture was cooled to 30 캜.

성분(중량%)Component (% by weight) 비교제형Comparative formulation 실험제형Experimental formulation 알부틴Arbutin -- 2.02.0 친유형 모노스테아린산글리세린Pro-type glycerin monostearate 2.02.0 2.02.0 세테아릴알콜Cetearyl alcohol 2.02.0 2.02.0 스테아린산Stearic acid 1.51.5 1.51.5 폴리솔베이트 60Polysorbate 60 1.51.5 1.51.5 솔비탄스테아레이트Sorbitan stearate 0.60.6 0.60.6 하이드로제네이티드 폴리이소부텐Hydrogenated polyisobutene 1.01.0 1.01.0 스쿠알란Squalane 3.03.0 3.03.0 광물유Mineral oil 5.05.0 5.05.0 사이클로메치콘Cyclomethicone 5.05.0 5.05.0 디메치콘Dimethicone 1.01.0 1.01.0 초산토코페롤Tocopheryl acetate 0.50.5 0.50.5 글리세린glycerin 5.05.0 5.05.0 베타인Betaine 3.03.0 3.03.0 트리에탄올아민Triethanolamine 1.01.0 1.01.0 산탄검Xanthan gum 0.050.05 0.050.05 방부제antiseptic 적량Suitable amount 적량Suitable amount 정제수Purified water 잔량Balance 잔량Balance 합계Sum 100100 100100

[피험자 선정][Selection of subjects]

18세~60세의 성인 남녀 중에서 평가 기관의 피험자 선정 기준에 만족하며, 선정 제외 기준에 해당되는 사항이 없는 사람을 피험자로 선정하였다. Among the adult males and females between 18 and 60 years old, the subjects were selected as the subjects who satisfied the evaluation criteria of the evaluation institution and did not meet the selection exclusion criteria.

평가자는 ‘평가자가 피험자에게 알려주는 사항’을 피험자에게 설명하였고, 피험자는 자의에 따라 ‘임상시험 참가 동의서’를 작성하고 실험에 참여하였다. The evaluator explained to the subject "the evaluator informed the subject", and the subject completed the "clinical study participation agreement" and participated in the experiment.

[화장품 도포][Cosmetic application]

누적 첩포 방법을 이용하여 비교제형과 실험제형의 화장품이 피부에 도포되도록 하였다. 첩포 테이프는 8mm 직경의 Finn chamber를 사용하여 피험자의 전완 또는 등 부위에 48시간 부착하였다. 이러한 첩포 테이프를 4주간 주 2회, 총 8회 적용하도록 하였다.
The cumulative spraying method was used to apply the comparative formulation and the cosmetic formulations to the skin. The patch tape was attached to the subject's forearm or back area using a Finn chamber of 8 mm diameter for 48 hours. This papermaking tape was applied 8 times a week for 4 weeks.

[각질층 채취 방법][Method for collecting stratum corneum]

피부로부터 첩포 테이프를 떼어낸 후, 각질층을 테이프 스트리핑 방법으로 박리 채취하였다. 채취 부위는 첩포 테이프에 의해 비교제형 및 실험제형이 도포된 부위로 하였다.
After removing the patch tape from the skin, the stratum corneum was peeled off by a tape stripping method. The sampling site was the area where the comparative formulation and the experimental formulation were applied by a patch tape.

[피부 각질층 중 단백질 추출 방법] [Protein Extraction Method in Skin Horny Layer]

각 각질층 테이프 1매에 대하여 500 ㎕의 T-PER Tissue Protein Extraction Reagent(Thermo Scientific 제조)를 넣고, 약 0.8 g의 유리 비드를 첨가하여 25분간 볼텍싱(vortexing)하여, 각질층을 파쇄하고, 각질층으로부터 단백질을 추출하였다.
PER Tissue Protein Extraction Reagent (manufactured by Thermo Scientific) was added to each of the stratum corneum tapes. 500 μl of T-PER Tissue Protein Extraction Reagent (manufactured by Thermo Scientific) was added and about 0.8 g of glass beads was added and vortexed for 25 minutes to crush the stratum corneum Proteins were extracted.

[피부 각질층 단백질의 정량][Determination of skin stratum corneum protein]

회수한 단백질 용해액의 단백질량을 Pierce BCA Protein Assay Kit(Thermo Scientific 제조)를 사용하여 정량하였다.
The protein amount of the recovered protein solution was quantitated using Pierce BCA Protein Assay Kit (manufactured by Thermo Scientific).

[멜라닌의 정량][Quantity of melanin]

상기 파쇄액으로부터, 멜라닌 양을 human melanin ELISA kit(Mybiosource 제조)를 사용하여 통상의 방법에 따라 측정하였다.
The amount of melanin was determined from the above-mentioned disruption solution by a conventional method using a human melanin ELISA kit (manufactured by Mybiosource).

[멜라닌 양 측정][Melanin amount measurement]

가. 비교제형 도포 부위의 멜라닌 양 측정end. Measurement of melanin level in comparative formulation application area

대조군 도포 부위에서 채취한 각질층에 존재하는 멜라닌 양을 도면 1에 나타내었다. The amount of melanin present in the stratum corneum collected from the control application region is shown in FIG.

나. 실험제형 도포 부위의 멜라닌 양 측정I. Measurement of melanin level at the application site

미백 화장품 도포 부위에서 채취한 각질층에 존재하는 멜라닌 양을 도면 2에 나타내었다. The amount of melanin present in the stratum corneum collected from the whitening cosmetic application region is shown in FIG.

다. 미백 화장품에 의한 피부 각질층의 멜라닌 억제 효능All. Effect of whitening cosmetics on melanin inhibition of skin stratum corneum

비교제형 도포 부위에서 측정된 멜라닌 양과 실험제형 도포 부위에서 측정된 멜라닌 양을 비교하여 화장품으로 인한 멜라닌 생성 억제 정도를 도 3에 나타내었다.The degree of inhibition of melanin production by cosmetics is shown in Fig. 3 by comparing the amount of melanin measured at the comparative dosage form application site with the amount of melanin measured at the application form application site.

이상의 결과로부터 실험제형으로 인하여 피부 각질층의 멜라닌 양이 감소한 것을 알 수 있었다. From the above results, it was found that the amount of melanin in the skin stratum corneum decreased due to the experimental formulation.

Claims (7)

화장료 조성물의 피부 미백 효과를 평가하는 방법으로서,
화장료 조성물 도포 전과 도포 후 피부 각질층의 멜라닌 양을 측정함으로써 화장료 조성물의 피부 미백 효과를 평가하는 방법. A method for evaluating the skin whitening effect of a cosmetic composition,
A method for evaluating the skin whitening effect of a cosmetic composition by measuring the amount of melanin in the horny layer of the skin before and after application of the cosmetic composition. 제1항에 있어서, 상기 화장료 조성물 도포는
누적 첩포 방법으로 수행하는 것을 특징으로 하는 평가 방법.The cosmetic composition according to claim 1, wherein the cosmetic composition is applied
And a cumulative deposition method. 제1항에 있어서, 상기 멜라닌 양은
효소 결합 면역 분석법, 방사면역분석법 또는 웨스턴 블로팅법을 통해 측정하는 것을 특징으로 하는 평가 방법.2. The method of claim 1, wherein the amount of melanin is
An enzyme-linked immunosorbent assay, a radioimmunoassay, or a Western blotting method. 제1항에 있어서, 상기 피부 각질층은 테이프 스트리핑에 의해 채취되는 것을 특징으로 하는 평가 방법.The evaluation method according to claim 1, wherein the skin stratum corneum is collected by tape stripping. 제1항에 있어서,
(S1) 피험자 각질층을 채취하여 각질층의 멜라닌 양을 측정하는 단계;
(S2) 피험자 피부에 시험 대상 화장료 조성물을 도포한 후 피부 각질층을 채취하여 각질층의 멜라닌 양을 측정하는 단계; 및
(S3) 상기 1) 단계 및 2) 단계에서 측정한 멜라닌양의 함량변화로부터 도포된 화장료 조성물의 피부 미백 효과를 평가하는 단계
를 포함하는 것을 특징으로 하는 평가방법. The method according to claim 1,
(S1) measuring the amount of melanin in the stratum corneum by taking a stratum corneum of the subject;
(S2) applying a test cosmetic composition to the testee's skin and collecting the skin stratum corneum to measure the amount of melanin in the stratum corneum; And
(S3) evaluating the skin whitening effect of the applied cosmetic composition from the change in the content of the melanin amount measured in steps 1) and 2)
And evaluating the evaluation result. 제 5항에 있어서, 상기 시험 대상 화장료 조성물은
비교제형 화장료 조성물과 평가하고자 하는 실험제형 화장료 조성물을 사용하는 것을 특징으로 하는 평가방법. 6. The composition according to claim 5, wherein the test cosmetic composition
Wherein the comparative formulation cosmetic composition and the experimental cosmetic composition to be evaluated are used. 제5항에 있어서, 상기 피부 미백 효과를 평가하는 단계는
비교제형 화장료 조성물과 실험제형 화장료 조성물 도포 전후 멜라닌 감소율 비교를 통해 수행하는 것을 특징으로 하는 평가방법. 6. The method of claim 5, wherein evaluating the skin whitening effect
Characterized in that the evaluation is carried out by comparing the reduction rate of melanin before and after application of the comparative cosmetic composition and the cosmetic composition for experimental formulation.
KR1020130159835A 2013-12-20 2013-12-20 Method for estimating skin whitening effect of cosmetic composition KR102134930B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
KR1020130159835A KR102134930B1 (en) 2013-12-20 2013-12-20 Method for estimating skin whitening effect of cosmetic composition

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1020130159835A KR102134930B1 (en) 2013-12-20 2013-12-20 Method for estimating skin whitening effect of cosmetic composition

Publications (2)

Publication Number Publication Date
KR20150072567A true KR20150072567A (en) 2015-06-30
KR102134930B1 KR102134930B1 (en) 2020-07-16

Family

ID=53518480

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1020130159835A KR102134930B1 (en) 2013-12-20 2013-12-20 Method for estimating skin whitening effect of cosmetic composition

Country Status (1)

Country Link
KR (1) KR102134930B1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20170014337A (en) 2015-07-29 2017-02-08 주식회사 바이오솔루션 Method for estimating skin whitening effect of material using human skin melanization model
KR20210087672A (en) 2020-01-03 2021-07-13 (주)아모레퍼시픽 Method for evaluating anti-aging performance of cosmetics and system performing the same
CN113718008A (en) * 2021-11-01 2021-11-30 深圳市萱嘉生物科技有限公司 Whitening raw material efficacy evaluation system and evaluation method

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102544440B1 (en) 2022-06-09 2023-06-20 주식회사 제이투케이바이오 A novel anti-wrinkle evaluation method that treats Enterobacter aerogenes J2K-739 strain derived from skin flora and Enterobacter genus derived from skin flora as an irritant
KR102544441B1 (en) 2022-06-09 2023-06-20 주식회사 제이투케이바이오 A novel anti-inflammatory evaluation method that treats Enterobacter aerogenes J2K-739 strain derived from skin flora and Enterobacter genus derived from skin flora as an irritant

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060020774A (en) * 2004-09-01 2006-03-07 한불화장품주식회사 A cosmetic composition for anti-aging containing an extract of melothria heterophylla
KR20070098073A (en) * 2006-03-31 2007-10-05 (주)아모레퍼시픽 Quantitative analyzing method of melanin in corneocyte to visually express the distribution of melanin in the skin surface
KR20090066824A (en) * 2007-12-20 2009-06-24 고려대학교 산학협력단 Compositions for skin whitening comprising puerarin or derivates thereof
KR20110134612A (en) * 2010-06-09 2011-12-15 한불화장품주식회사 A skin-care agent containing ophioglossum vulgatum extracts using microbial fermentation
KR20130082063A (en) * 2012-01-10 2013-07-18 바이오스펙트럼 주식회사 Agents for skin whitening containing artemisinic acid, its derivatives and pharmaceutically acceptable salts thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060020774A (en) * 2004-09-01 2006-03-07 한불화장품주식회사 A cosmetic composition for anti-aging containing an extract of melothria heterophylla
KR20070098073A (en) * 2006-03-31 2007-10-05 (주)아모레퍼시픽 Quantitative analyzing method of melanin in corneocyte to visually express the distribution of melanin in the skin surface
KR20090066824A (en) * 2007-12-20 2009-06-24 고려대학교 산학협력단 Compositions for skin whitening comprising puerarin or derivates thereof
KR20110134612A (en) * 2010-06-09 2011-12-15 한불화장품주식회사 A skin-care agent containing ophioglossum vulgatum extracts using microbial fermentation
KR20130082063A (en) * 2012-01-10 2013-07-18 바이오스펙트럼 주식회사 Agents for skin whitening containing artemisinic acid, its derivatives and pharmaceutically acceptable salts thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20170014337A (en) 2015-07-29 2017-02-08 주식회사 바이오솔루션 Method for estimating skin whitening effect of material using human skin melanization model
KR20210087672A (en) 2020-01-03 2021-07-13 (주)아모레퍼시픽 Method for evaluating anti-aging performance of cosmetics and system performing the same
CN113718008A (en) * 2021-11-01 2021-11-30 深圳市萱嘉生物科技有限公司 Whitening raw material efficacy evaluation system and evaluation method

Also Published As

Publication number Publication date
KR102134930B1 (en) 2020-07-16

Similar Documents

Publication Publication Date Title
Del Bino et al. Clinical and biological characterization of skin pigmentation diversity and its consequences on UV impact
Buhé et al. Pathophysiological study of sensitive skin
Nam et al. The seasonal variation in skin hydration, sebum, scaliness, brightness and elasticity in Korean females
KR101199672B1 (en) Methods of treating skin with aromatic skin-active ingredients
KR20150072567A (en) Method for estimating skin whitening effect of cosmetic composition
Luebberding et al. Age-related changes in male skin: quantitative evaluation of one hundred and fifty male subjects
US20090068219A1 (en) Anti-wrinkle hormone-type cosmetic composition
Naik et al. Influence of ethnicities and skin color variations in different populations: a review
JP5582630B2 (en) Cosmetic composition containing an extract from lotus and method of cosmetic care using said composition
Draelos et al. A re-evaluation of the comedogenicity concept
Mangelsdorf et al. Ethnic variation in vellus hair follicle size and distribution
JP5275898B2 (en) Evaluation method of skin condition
Jeong et al. Comparison of the efficacy of Atopalm® multi-lamellar emulsion cream and Physiogel® intensive cream in improving epidermal permeability barrier in sensitive skin
Addor Topical effects of SCA®(Cryptomphalus aspersa secretion) associated with regenerative and antioxidant ingredients on aged skin: evaluation by confocal and clinical microscopy
CN103402494A (en) Methods for improving the appearance of aging skin using a composition comprising banyan tree, lotus, and clover serum fractions
US10175230B2 (en) Use of biomarkers for evaluating the effectiveness of active ingredients
JP2024026167A (en) Diagnostic kit of minimally invasive skin condition including microneedle patch
Williams et al. Gender difference of in vivo skin surface pH in the axilla and the effect of a standardized washing procedure with tap water
JP2023062016A (en) Agents for improving skin barrier function worsened due to stress
Kaur et al. Skin care assessment on the basis of skin hydration, melanin, erythema and sebum at various body sites
JP2014163715A (en) Method for evaluating resistance to ultraviolet damage
Khan et al. Hippophae rhamnoides oil-in-water (O/W) emulsion improves barrier function in healthy human subjects
KR102201385B1 (en) Whitening cosmetic compositions and whitening cosmetics using the same
WO2020117550A1 (en) White pine bark extract for decreasing endothelin-1 secretion, stem cell factor synthesis and protein carbonylation
Lane et al. The presence of essential and non-essential stratum Corneum proteases: the vital need for protease inhibitors

Legal Events

Date Code Title Description
A201 Request for examination
E902 Notification of reason for refusal
E701 Decision to grant or registration of patent right