WO2012008747A2

WO2012008747A2 - Skin diagnosis method and kit

Info

Publication number: WO2012008747A2
Application number: PCT/KR2011/005147
Authority: WO
Inventors: 김신형; 오택진; 김형준; 손의동; 강병영
Original assignee: (주)아모레퍼시픽
Priority date: 2010-07-13
Filing date: 2011-07-13
Publication date: 2012-01-19
Also published as: WO2012008747A3; KR101409610B1; KR20120006945A

Abstract

The present invention relates to a method for diagnosing skin conditions, comprising: a step of collecting skin or a skin secretion; and a step of measuring a biomarker of the skin or skin secretion collected in the previous step. The present invention also relates to a kit for diagnosing skin conditions, comprising: a unit for collecting skin or skin secretions; and a measuring unit which measures a biomarker of the skin or skin secretion. Skin conditions can be easily, conveniently, and quickly measured using the above-described method for diagnosing skin conditions and the kit for diagnosing skin conditions.

Description

【Specification】

[Name of invention]

Skin Diagnostic Methods and Kits

Technical Field

The present invention relates to a skin diagnostic method and a skin diagnostic kit.

Background Art

<2> Bio marker refers to a general substance used as an indicator of a biological state. It is mainly used as an objective indicator in the medical field to know whether a person is in a healthy or diseased state, and the degree of reaction to drug treatment. The biomarker can be used not only in the medical field but also in various fields. In detail, the skin care field may be used as an indicator of skin condition.

[Detailed Description of the Invention]

[Technical problem]

<3> The present invention comprises the steps of collecting the skin or skin secretions to diagnose the skin condition easily, simply and quickly and accurately; And to provide a skin condition diagnostic method comprising the step of measuring the biomarker (bio marker) of the skin or skin secretion collected above. In addition, the present invention skin or skin secretion collection unit; And to provide a skin condition diagnostic kit comprising a bio marker (bio marker) measuring unit of the skin or skin secretions.

Technical Solution

<4> one aspect of the present invention comprises the steps of collecting the skin or skin secretions; And it provides a skin condition diagnostic method comprising the step of measuring the biomarker of the skin or skin secretion collected above.

Another aspect of the present invention is the skin or skin secretion collection unit; And it provides a skin condition diagnostic kit comprising a biomarker measuring unit of the skin or skin secretions.

Advantageous Effects

Anyone can easily, easily and quickly diagnose a skin condition at any place by using a skin condition diagnosis method and a skin diagnosis kit according to an aspect of the present invention. Furthermore, based on the diagnosis results, each individual can receive the skin care that is appropriate for them.

[Brief Description of Drawings]

<7> Figure 1 is a reaction containing the solution containing lactate hydrogenase and sebum collected It is a photograph showing the figure.

2 shows polyvinyl alcohol (PVA) and polyoxyethylene sorbitan monooleate, respectively.

(PSM) and ethane are photographs showing the spreadability and absorption of a solution containing (EtOH).

FIG. 3 is a photograph showing the reactivity of a solution containing lactate hydrogenase and polyoxyethylene sorbitan monooleate (PSM) by concentration.

Figure 4 is a photograph showing the reaction properties of the solution containing lactate hydrogenase by concentration.

5 is a photograph showing the reactivity of a solution containing lactate hydrogenase and distilled water or polyoxyethylene sorbitan monooleate.

[Form for implementation of invention]

As used herein, the term "skin" refers to a tissue covering the body surface of an animal, and is a broad concept including not only tissues covering the body surface such as the face or body, but also the scalp and hair.

<13>

<14> Hereinafter, this invention is demonstrated in detail.

Since a skin type and a skin condition are different for each person, diagnosis of the skin condition of the person must be preceded in order to achieve proper skin care for each individual. Currently used skin diagnosis methods include skin diagnosis through a questionnaire and skin diagnosis using a bulky skin diagnosis device. Among them, a skin diagnosis through a questionnaire is relatively simple and can be made without restriction of place, but it has a disadvantage of being unscientific. Although skin diagnosis using a bulky skin diagnosis device can obtain relatively accurate results, the diagnosis can be made only where the device is located, which places limitations and in most cases it takes a long time to obtain a result. have. Therefore, if skin diagnosis can be made more easily, quickly and accurately, each individual will not only be able to diagnose the skin conveniently, but also be able to receive appropriate skin care based on the diagnosis result.

<! 6>

Uniaxial surface of the present invention comprises the steps of collecting the skin or skin secretions; And it provides a skin condition diagnostic method comprising the step of measuring the biomarker of the skin or skin secretion collected above.

In the method of diagnosing the skin condition according to an aspect of the present invention, the skin or skin secretion is collected at a stage such as a patch or oil paper. One or more of paper, film, and stripping tape can be used. Specifically, the skin or skin secretions may be removed by attaching and detaching the patch, paper or film on the skin for a predetermined time, that is, for 5 minutes to 10 minutes, for example, after the skin or skin secretion is collected. Can be harvested. Alternatively, the skin or skin secretion may be collected by attaching and stripping the stripping tape to the skin for a predetermined time, that is, for 1 hour, for example, from 10 seconds to 7 minutes, for a period of time to extract the skin or skin secretions.

One side of the present invention is a skin or skin secretion collecting unit; And it provides a skin condition diagnostic kit comprising a biomarker measuring unit of the skin or skin secretion. In the kit for diagnosing skin conditions according to an aspect of the present invention, the collecting unit may include one or more of a patch, paper, film, and a stripe tape.

The kit for diagnosing skin condition according to an aspect of the present invention may include a skin or skin secretion collecting unit and a biomarker measuring unit of skin or skin secretion in an integrated form or in an isolated form. When the skin condition diagnosis kit according to an aspect of the present invention includes a skin or skin secretion collecting unit and a biomarker measuring unit in an integrated form, the skin or skin secretion collecting unit and the biomarker measuring unit of the skin condition diagnosis kit are patched. Or in the form of a stripping tape. When the kit for diagnosing a skin condition according to an aspect of the present invention includes a skin or skin secretion collecting unit and a biomarker measuring unit in a separate form, the skin or skin secretion collecting unit includes a stripping tape and the biomarker measuring unit is a chip. To sensor form.

<21>

In one aspect of the present invention, the skin condition includes at least one of skin aging degree, skin moisturizing degree, skin oxidation degree, skin stress level, skin oil level, skin keratin degree, skin sensitivity, skin color, and skin inflammation level. do. In another aspect of the invention, the skin condition includes one or more of the degree of skin aging, the degree of skin moisturizing and the degree of skin oxidation. Meanwhile, the degree of skin oxidation may mean city stress.

The skin condition diagnosis kit according to an aspect of the present invention includes one or more of the biomarkers for diagnosing the skin condition. In another aspect of the present invention, the biomarker for diagnosing the skin condition may include all of the biomarkers generally used in the art for diagnosing each skin condition. In one aspect of the present invention, a biomarker for diagnosing skin hydration is cathepsin L, invohicrin, skin pH, SC stratum corneum neutral lipids, SCCstratum corneum Serine proteases (e.g., SC tryptase-like enzymes, plasmin, urokinase, trypsin-like kallikreins (KL 5) or chymoses) Trypsin-like kallikreins (KLK7), transglutaminase (TGase), desmogrine Kdesmoglein 1), loricrin, fi laggrin, lamellar body), bleomycin hydrolase, calpain, caspase-14, desmoglein 3, PLSCR, SPRR IB or 2B, lactate ), Coagulase and stratum corneum carbo Chemistry and proteins containing one or more selected from the group consisting of (stratum corneum car bony 1 ated protein, SCCP).

In one aspect of the invention, the biomarker for diagnosing the degree of skin aging is IL-βΐ, IL-6, 匪 P-1, 顧顧 -8, 雇 P-13, TNF-α, collagen I, Collagen ΙΠ, hyaluronic acid, fibronectin, keratin 1, keratin 10, gellatinase-mmp 2,9 and stratum corneum carbonylated protein (SCCP) It includes one or more selected from the group consisting of. In one aspect of the present invention, the biomarker for diagnosing skin color may include MC1R, endothelin receptor, tyrosinase, TRP 1, TRP 2, MITFCmi crophthalmi a-assoc i ated transcription factor. , g LOO, Marti, inter leukin-6, prostaglandin, leukotr iene, SCF, Vascular endothelial growth factor, VEGF, c-kit, EGF-receptor (epidermal growth) one or more selected from the group consisting of factor-receptor, melanin and stratum corneum carbonylated protein (SCCP).

In one aspect of the invention, the biomarker for diagnosing the degree of skin oxidation is

TRX (thioredoxin) 1, TRX 2, Prx (peroxiredoxin) 1, 2, 3, 4, 5 or 6, Grx (glutaredoxin) 1, Grx 2, TrxRKTrx reductase), TrxR2, NQOl, Nrf2, catalase At least one selected from the group consisting of glutathione and cortisol.

In one aspect of the invention, the biomarker for diagnosing skin sensitivity includes coagulase. More specifically, in one aspect of the invention, the biomarker for diagnosing the degree of skin aging is at least one of gelatinase and stratum corneum car bony 1 at ed protein (SCCP) It includes. In another aspect of the invention, biomarkers for diagnosing skin hydration include stratum corneum carbonylated protein (SCCP), lactate, H, stratum corneum tryptase-like enzyme (SC). tryptase-like enzyme, plasmin, trypsin-like kallikreins (KLK5), chymotrypsin-like kallikreins (KLK7) and transglutaminase, TGase). In another aspect of the present invention, the biomarker for diagnosing the degree of skin oxidation includes at least one selected from the group consisting of catalase, glutathione, and cathepsin LCcathepsin L).

More specifically, in one aspect of the present invention, the biomarker for diagnosing a skin condition includes lactate. Lactate is a natural moisturizing factor (NMF), which accounts for the largest amount (about 12%) after amino acids and ions. Lactate promotes ceramide biosynthesis, strengthens skin barrier function and affects skin moisture, skin pH and skin stiffness (J Invest Dermatol. 2004 Mar; 122 (3): 755-763). Therefore, when the lactate degree in the skin is high, a skin condition that can be exemplified as the degree of skin moisturizing can be diagnosed as preferable.

<31>

In one aspect of the invention, the skin or skin secretion comprises skin keratin. In general, the biomarkers, which may include lactate, are known to exist in the stratum corneum. Therefore, the skin condition can be diagnosed by collecting the skin keratin and measuring the amount of biomarkers such as lactate contained therein.

In another aspect of the invention, the skin or skin secretion comprises sebum. To get keratin from facials, face-washed women should wash their face. This is a very cumbersome process, and since it is known that the amount of lactate in keratin decreases after cleansing, face washing may be an obstacle to accurate diagnosis. Skin secretions, such as sebum, can be collected easily and painlessly without washing. Since the amount of lactate in the dead skin cells and skin secretions is similar to each other, the skin condition is measured by measuring the amount of biomarkers such as lactate contained in the skin secretions. Can be diagnosed. In another aspect of the invention, the skin or skin secretion is the skin of the face, specifically the face T zone or Contains skin secretions.

<34>

In general, when diagnosing a skin condition, skin keratin is collected, the collected skin keratin is extracted with water or SDS buffer, and then absorbance is measured and compared with a standard curve to quantify the amount of biomarker and based on the results. Diagnose skin conditions. This conventional method is cumbersome, time consuming and expensive. On the other hand, the method for diagnosing skin condition according to an aspect of the present invention reacts a composition comprising at least one selected from the group consisting of enzymes, substrates and surfactants, that is, reaction solutions and biomarkers in skin or skin secretions without extraction and quantification processes. By measuring the degree of reaction of the biomarker to the composition, the concentration or amount of the biomarker can be measured. As described above, the method for diagnosing skin condition according to an aspect of the present invention can easily and efficiently diagnose the skin condition without undergoing an extraction and quantification process. The biomarker measuring unit of the skin condition diagnosis kit according to an aspect of the present invention includes at least one selected from the group consisting of enzymes, substrates and surfactants, and reacts biomarkers with at least one of enzymes, substrates and surfactants. By measuring the degree of response, the skin condition can be diagnosed immediately and easily.

In one aspect of the invention, the enzyme or substrate may be a pair of enzymes or mating substrates that specifically react with the biomarker to be measured. In another aspect of the invention, the enzyme or substrate can be determined according to the biomarker to be measured. In another aspect of the invention, when the biomarker to be measured is lactate, the corresponding enzyme reacts specifically with lactate to produce lactate dehydrogenase (Lactate) to produce pyruvate. Dehydrogenase).

In one aspect of the invention, the composition comprises an enzyme or substrate having a concentration of greater than 0.05 mg / ml, specifically 0.2 mg / ml or more, 10 mg / ml or less, specifically 5 mg / ml or less. can do. Including the concentration in the above range is not only suitable for showing the intended effect of the present invention, but also satisfies both the stability and safety of the composition, it may be appropriate to include the concentration in the above range in terms of cost-effectiveness. . Specifically, when using the enzyme of 0.05 mg / ml takes a reaction time 30 minutes or more, in one aspect of the present invention can be completed within 20 minutes by using an enzyme of a concentration higher than 0.05 mg / ml The skin condition can be diagnosed quickly. In addition, when the enzyme is used in a concentration higher than 0.05 mg / ml increases the reaction properties can be easily determined by the naked eye. In one aspect of the present invention, the composition that reacts with the biomarker includes a surfactant, thereby increasing the spreading area to increase the contact area with the biomarker, and improve the absorption properties of the biomarker by improving the spreadability. You can. In another aspect of the invention, the surfactant is a polyoxyethylene sorbitan monooleate

(Polyoxy ethylene Sorbitan Mwiool eat e) (t ween 80 tween 60 ^ tween 40 ^⑧ or tween 20 ^

In one aspect of the invention, the composition is a concentration of 0.1% or more, specifically 0.5% or more, more specifically 5% or more, less than 100%, specifically 20% or less, more specifically 10% or less. The branch may comprise a surfactant. Including the concentration in the above range is not only suitable for showing the intended effect of the present invention, but also satisfies both the stability and safety of the composition, it may be appropriate to include the concentration in the above range in terms of cost-effectiveness.

In one aspect of the invention, the biomarkers measurable by enzymes or substrates are gelatinase, stratum corneum tryptase-like enzymes, trypsin-like kallikrein (KLK5), chymotrypsin-like kallikrein (KLK7), plasma One or more of Min and Cathepsin L.

In one aspect of the invention, biomarkers measurable by enzymes or substrates and surfactants include one or more of lactate, catalase, glutathione and transglutaminase (TGase).

<42>

In the method for diagnosing skin condition according to an aspect of the present invention, the step of measuring the biomarker of the skin or skin secretion is to measure the response of the biomarker to the reagent by reacting the biomarker and the reagent. The process may further include. The reagent may include a chemical material that displays the reaction composition and the degree of reaction of the biomarker in fluorescence or color, which may be determined according to the biomarker to be measured, and thus an enzyme or substrate. In another aspect of the present invention, the reagent may be present in a separate reaction composition or present separately. The kit for diagnosing skin condition according to an aspect of the present invention may further include a reagent for measuring the biomarker of the skin or skin secretion collected.

<44>

In the step of measuring the biomarker of the method for diagnosing skin condition according to an aspect of the present invention and the biomarker measuring unit of the kit for diagnosing skin condition, the biomarker measurement may be performed by fluorescence assay or colormetric assay. Can be All. Specifically, the degree of response of the biomarker to the reaction composition may be measured by fluorescence or color appearing after the collected biomarker is reacted with the reaction composition for a predetermined time.

In one aspect of the invention, biomarkers measurable using fluorescence quenching methods include gelatinase, stratum corneum carbonylated protein (SCCP), cathepsin L, stratum corneum tryptase-like enzymes, plasmin, trypsin-like. One or more of kallikrein (KLK5) and chymotrypsin-like kallikrein (KLK7). In another aspect of the invention, biomarkers measurable using colorimetric methods include one or more of lactate, catalase, glutathione and transglutaminase (TGase).

<47>

In the method for diagnosing skin condition according to an aspect of the present invention, the biomarker measurement result may be displayed in fluorescence or color. The indicated fluorescence or color can be confirmed by fluorescence analyzer or naked eye.

In the kit for diagnosing skin conditions according to an aspect of the present invention, the biomarker measuring unit may include a fluorescence or color display unit, and the fluorescence or color display unit may display whether or not the measured biomarker is in fluorescence or color. have. Specifically, the biomarker and the substrate may react for a predetermined time, and the reaction may be displayed in fluorescence or color on the fluorescence or color display unit.

In one aspect of the present invention, the degree of fluorescence or color display may vary according to the degree of the measured biomarker. In another aspect of the present invention, the measured biomarker may have a high degree of fluorescence or color intensity, which may appear strongly or weakly. In another aspect of the present invention, a plurality of fluorescent or color indicators are present in the biomarker measuring unit, each biomarker in the plurality of biomarker measuring unit has a different concentration, the biomarker according to the skin condition The number of fluorescence or color indicators displayed on the measurement unit may vary, and through this, the skin condition may be determined. For example, two or more fluorescence or color display units exist, and as the concentration of the measured biomarker increases, the number of fluorescence or color display units for displaying fluorescence or color may increase.

In one aspect of the present invention, the fluorescence or color display unit may display fluorescence or color according to the degree of the measured biomarker. In another aspect of the present invention, when the measured biomarker is in a normal range, fluorescence or color may not be displayed. In another aspect of the invention, the degree of biomarker measured is not within the normal range. In the case of fluorescence, fluorescence or color may be indicated. The term "degree" of the biomarker may mean the concentration or amount of the biomarker.

In the uniaxial aspect of the present invention, the skin condition can be diagnosed according to whether or not fluorescence or color display. In another aspect of the present invention, it is possible to diagnose that the skin condition is preferable or undesirable as the fluorescence or color is not displayed or the display intensity is weak. The higher the intensity, the more likely the diagnosis is that skin conditions are desirable or undesirable. In another aspect of the present invention, after comparing the measured intensity with the normal display intensity classified by sex or age, based on the results it can be diagnosed that the skin condition of the subject is preferable or undesirable.

In another aspect of the present invention, when the biomarker to be measured is gelatinase or stratum corneum carbonylated protein (SCCP), it is diagnosed that aging of the skin is likely to be fluorescence or color, and its intensity may be strong. can do. In another aspect of the invention, the biomarkers measured are stratum corneum carbonylated protein (SCCP), pH, stratum corneum tryptase-like enzymes, plasmin, trypsin-like kallikrein (KLK5) and chymotrypsin-like kallikrein In the case of one or more of (KLK7), it can be diagnosed that the fluorescence or color is displayed and the intensity of the skin is lower as the intensity is stronger. In another aspect of the present invention, when the biomarker to be measured is lactate or transglutaminase (TGase), it may be diagnosed that the fluorescence or color is displayed and the intensity of the skin is higher as the intensity thereof is stronger. In another aspect of the present invention, when the biomarker to be measured is cathepsin L, it may be diagnosed that the degree of skin oxidation is higher as fluorescence or color is displayed and the intensity thereof is stronger. In another aspect of the present invention, when the biomarker to be measured is glutathion or catalase, it may be diagnosed that fluorescence or color is displayed and the intensity of skin oxidation is low. That is, the relationship between whether fluorescence or color is displayed, and its intensity and skin condition, may vary depending on the biomarker to be measured.

<54>

The kit for diagnosing skin condition according to an aspect of the present invention may further include a reader capable of observing fluorescence or color change. In another aspect of the invention, the reader may be a portable fluorescence image application (Fluorescence image application) capable of measuring the fluorescence.

According to another aspect of the present invention, a kit for diagnosing a skin condition includes instructions for using the kit, an indication including at least one selected from a skin condition indicator and a skin care method. It may further include.

The skin condition diagnosis method according to one aspect of the present invention may be performed using a skin condition diagnosis kit according to another aspect of the present invention.

<58>

When using the method for diagnosing skin condition and kit for diagnosing skin condition according to an aspect of the present invention, scientific skin condition diagnosis using an objective indicator such as a biomarker can be quickly obtained within about 20 minutes. Non-experts can also easily and easily diagnose skin conditions. Furthermore, based on the result of diagnosis of the skin condition, each individual may select a cosmetic and skin care method suitable for him or her.

<60>

Experimental Example 1 Comparison of Biomarker Levels of Skin Keratin and Sebum

(1) Comparison of the amount of biomarkers of keratin according to washing

In general, biomarkers for diagnosing skin conditions are known to exist in the stratum corneum. In order to harvest keratin, it is usually necessary to clean the skin first. Hereinafter, we examined whether washing could be an obstacle to the measurement of the amount of biomarkers in the skin keratin of lactate among the biomarkers.

<64>

<65> Applying stripping tape (D-Squam, obtained from CuDerm) for 10 seconds to each part of a man and woman 20 years or older without washing one forehead and the other forehead without washing Keratin was collected by. Each striping tape was placed in a 5% SDS-buffer solution for reaction for 30 minutes and then centrifuged. Sample 50 / ^ obtained from each stripping tape and reaction sample 50 ^ containing lactate substrate and enzyme were placed in a reaction vessel, and the reaction was carried out for 30 minutes, followed by dark reaction, and the O.D. was measured at 450 nm. Measured O.D. The average of the values is shown in the table below (significant 95% confidence level).

<66> [Table 1]

As can be seen from the above results, the amount of lactate without washing was higher than that with washing. Thus, it can be seen that skin cleansing reduces the amount of biomarkers in the keratin. For women who are makeup, cleaning is required to collect dead skin cells Therefore, there is a need for a method for accurately measuring the amount of biomarker without cleaning.

<69>

(2) Comparison of the amount of biomarkers of skin keratin and sebum

Unlike the exfoliation of skin, sebum collection is convenient because it does not need to be cleaned in advance even when makeup is applied, and pain is less when collected. Therefore, if the amount of biomarkers of sebum and the amount of biomarkers of keratin are within the same range, skin condition can be diagnosed by collecting sebum and measuring the amount of biomarkers thereof. Hereinafter, the amount of biomarkers of skin keratin and sebum was compared with lactate among biomarkers.

<72>

<73> Keratin from 20 years old and older is collected with D-Squain stripping tape for skin exfoliation on one side of the forehead and obtained from oil blotting film, Sebutape, Cuderm on the other side. Sebum was collected). Each sample was placed in a 5% SDS-buffer solution, reacted for 30 minutes, and then centrifuged. After the reaction ∞ μί and the lactate group ¾ effect in the reaction vessel were put into each reaction, the reaction was performed for 30 minutes, and the O.D. was measured at 450 nm. Measured O.D. The results of quantifying the lactate amount compared to the standard curve are shown in the table below (significant 95% confidence level).

<74> [Table 2]

<75>

As can be seen from the above results, sebum lactate content was higher than keratin, and the P value of both was not different. That is, when sebum * is collected and the biomarker is measured, the results reflecting the current skin condition as well as or more than the case of measuring the biomarker in the skin keratin can be obtained. In addition, sebum collection as described above is easy, there is an advantage that less pain in the collection.

<77> Experimental Example 2 Evaluation According to Sebum Collection Method

<79> Which method of using sebum collecting film or oil-blotting paper when sebum is collected to diagnose skin condition can obtain a diagnosis result that better reflects current skin condition? To experiment was performed as follows.

<80> sebum film for forehead sebum for adults over 20 years old (Sebutape,

Sebum was collected by attaching and removing oil paper (obtained from CuDerm) and oil paper (obtained from AMOREPACIFIC) for 5 minutes and detaching them. Each was poured into 5% SDS-buffer solution and reacted for 30 minutes before centrifugation. In a reaction vessel, a sample 50 obtained from a sebum collecting film and an oil paper and a reaction solution 50 ^ containing a lactate substrate and lactate dehydrosenase were put in a dark reaction for 30 minutes, and the O.D. was measured at 450 nm. Measured O.D. The result of quantifying the lactate amount by comparing the value with the standard curve is shown in the table below.

<81> [Table 3]

<82>

As can be seen from the above results, it was confirmed that the sebum collected with the sebum collecting film and the oil paper contained similar amounts of lactate. That is, when sebum is collected using a sebum collecting film or oil paper and the skin condition is diagnosed, it can be seen that both of the results reflecting the current skin condition well.

<84>

Experimental Example 3 Evaluation According to Reaction Solution

(1) Method using reaction solution

In general, when diagnosing a skin condition, the collected skin is extracted with an SDS buffer, and then absorbance is measured and compared with a standard curve to quantify the amount of the biomarker and diagnose the skin condition based on the result. This conventional method is cumbersome, time consuming and expensive. Therefore, the present inventors have tested whether the skin condition can be diagnosed by a simple method of reacting the reaction composition with the skin biomarker without an extraction process. <88>

<& 9> A sebum-collecting film (obtained from Sebutape, CuDerm) was attached to the sebum-bearing Τ zone of three men and women aged 20 and over (experimental group λ, Β, (：)) for 5 minutes, and then detached. The film was placed in a reaction vessel containing a reaction solution containing a substrate and lactate dehydrogenase, and then stirred for 30 minutes, after which a photograph of the reaction vessel is shown in FIG.

As shown in FIG. 1, the color development due to lactate reaction was confirmed in all of the experimental groups A, B, and C, which collected sebum, which was not observed in the control group, which was not sebum collected. This is also the same result that can be obtained after extracting the extracted skin with SDS-buffer. In other words, it can be seen that the skin can be diagnosed easily and quickly without the process of extracting the seed and SDS-buffer by reacting the collected skin and the reaction solution.

<91>

(2) Evaluation according to the reaction solution components

In one aspect of the present invention, the skin biomarker and the reaction mixture are reacted to measure the degree of reaction of the biomarker with respect to the composition to diagnose the skin condition. At this time, if the spreadability of the reaction composition is high, the contact area with the biomarker becomes wider, and the reaction properties with the biomarker can be improved, and even when the reaction properties with the biomarker are high, This results in more accurate diagnostic results. In the following, the biomarker is a lactate as a representative example. Through experiments to change the components of the reaction composition, it was examined whether reactivity with the biomarker can be improved when the reaction component contains any component.

<94>

10% polyvinyl alcohol (PVA), ethanol (EtOH), and polyoxyethylene sorbitan monooleate (PSM, Tween 2 (?) Were prepared to prepare a film for sebum extraction (3Μ Oil blotting film, obtained from 3M), and the spreadability and water absorption were observed after 15 and 30 minutes, and the results are shown in FIG.

As shown in FIG. 2, the solution containing polyoxyethylene sorbitan monoacrylate of 1 showed the best spreadability and water absorption. That is, it can be seen that polyoxyerylene sorbitan monoacrylate is most excellent in terms of spreadability and water absorption.

<97>

(3) Evaluation of influence by polyoxyethylene sorbitan monoacrylate When the biomarker is lactate, a reaction solution containing lactate dehydrogenase, which may be used as a corresponding enzyme, and 0.1%, 0.5%, and 1¾ polyoxyethylene sorbitan monooleate, respectively, is prepared and sebum Droplets were placed on a sample film (3M Oil blotting film, obtained from 3M), and after 30 minutes, the reactivity, spreadability and absorbency were observed. The results are shown in FIG.

<ιοο> As can be seen in Figure 3, polyoxyethylene sorbitan mono acrylate shows a significant color development at all concentrations do not affect the reactivity of lactate, and also shows a significant spread and hop number at all have.

<101>

From the above results, it can be seen that when the reaction solution contains a polyoxyethylene sorbitan monooleate as a surfactant, it has excellent reactivity, spreadability, and absorption, and thus, improved skin condition diagnosis can be obtained.

<103>

Experimental Example 4 Evaluation According to Enzyme Concentration

In one aspect of the invention, the skin condition is diagnosed by measuring the degree of reaction of the biomarker to the composition by reacting the reaction composition comprising the skin biomarker and the enzyme. Therefore, it was evaluated how the degree of reaction of the biomarker for the composition varies according to the enzyme concentration.

<) 06>

(1) Evaluation by Enzyme Concentration

After preparing a solution containing 0.01 mg / ml, 0.05 mg / ml, 0.1 mg / ml, and 0.2 mg / ml lactate dehydrogenase, respectively, the sebaceous film (3M Oil blotting film, 3M Getting a room on top of each other and observed the reaction after 30 minutes. The results are shown in FIG. As can be seen in Figure 4, it can be seen that the solution containing 0.2 mg / ml lactate dehydrogenase shows the most intense colorolol has the most excellent reaction. In addition, the use of 0.2 mg / ml lactate dehydrogenase can be obtained quickly reaction results, it is easy to observe the results can be observed with the naked eye.

That is, it can be seen that the improved skin condition diagnosis can be obtained quickly and easily when using 0.2 mg / ml of enzyme rather than 0.05 mg / ml of enzyme.

<110> <πι> (2) Application to reaction solution

<Π2> Banung solution containing 0.05 mg / ml and 0.2 mg / ml lactate dehydrogenase and distilled water, and 0.2 mg / ml lactate dehydrogenase and 0. A reaction solution containing non-elastic monooleate (PSM, Queen 2 (Λ) was prepared.

On the sebaceous film (Sebutape, obtained from CuDerm) of four semen and men sebum of 20 years old or older, the prepared reaction solution was placed one by one, and the reactivity was observed after 30 minutes. The results are shown in FIG. 5 (number at the top of the figure: subject identification number, A: lactate dehydrogenase and distilled water of 0.05 mg / ml, B: lactate dehydrogenase and distilled water of 0.2 mg / ml, C: 0.2 mg / ml lactate dehydrogenase and 0.1% polyoxyethylene sorbitan monoacrylate). As can be seen in FIG. 5, 0.2 mg / ml lactate dehydrogenase and 0.1% polyoxyethylene sorbitan monooleate (PSM,

It was found that the reaction solution containing tween 20 ^ showed the strongest color development and the highest reactivity.

<113>

From the above results, it can be seen that the improved skin condition diagnosis results can be obtained quickly and easily when using a reaction solution containing a surfactant and a higher concentration of enzyme than the conventional one.

Claims

[Range of request]

[Claim 1]

Collecting skin or skin secretions; And

A skin condition diagnostic method comprising the step of measuring a bio marker (skin biomarker) of the skin or skin secretion collected above.

[Claim 2]

The method of claim 1,

A skin condition is a method of diagnosing a skin condition that is one or more of skin aging, skin hydration, and skin oxidation.

[Claim 3]

The method of claim 2,

Biomarkers measured to diagnose the degree of skin aging include at least one of gelatinase and stratum corneum car bony 1 at ed protein (SCCP);

Biomarkers measured to diagnose skin hydration include stratum corneum car bony 1 at ed protein (SCCP), lactate, pH, and stratum corneum tryptase-like enzymes (SC tryptase-like). enzyme, plasmin, trypsin-like kallikreins (KLK5), chymotrypsin-like kallikreins (KLK7) and transglutaminase (TGase) One or more selected from the group;

The biomarker measured to diagnose the degree of skin oxidation comprises at least one selected from the group consisting of catalase, glutathione and cathepsin cathepsin L.

[Claim 4]

The method of claim 1,

A skin or skin secretion is a method of diagnosing a skin condition that involves one or more

[Claim 5]

The method of claim 1,

In the step of collecting the skin or skin secretion, the collection is a skin condition diagnosis method using one or more of a patch (patch), paper, film (striping tape).

[Claim 6]

The method of claim 1,

In the step of measuring the biomarker, after reacting the biomarker with a reaction composition comprising at least one selected from the group consisting of enzymes, substrates and surfactants to measure the degree of resilience of the biomarker to the composition,

The enzyme or substrate is a method for diagnosing skin conditions, which is an enzyme or substrate specifically reacting with the biomarker to be measured.

[Claim 7]

The method of claim 1,

In the step of measuring the biomarker, biomarker measurement is a skin condition diagnosis method using fluorescence or colorimetric method.

[Claim 8]

Skin or skin discharge portion; And

Skin condition diagnosis kit comprising a bio marker measurement unit of the skin or skin secretion (bio marker).

[Claim 9]

The method of claim 8,

A skin condition diagnosis kit wherein the skin condition is at least one of skin aging, skin hydration, and skin oxidation.

[Claim 10]

The method of claim 9,

Biomarkers for diagnosing degree of skin aging include one or more of gelatinase and stratum corneum carbonylated protein (SCCP);

Biomarkers for diagnosing skin hydration include stratum corneum carbonylated protein (SCCP), lactate, H, SC tryptase-like enzyme and plasmin ( one or more selected from the group consisting of plasmin, trypsin-like kallikreins (KLK5), chymotrypsin-like kallikreins (KLK7), and transglutaminase (TGase) It includes;

The biomarker for diagnosing skin oxidation is one selected from the group consisting of catalase, glutathione and cathepsin LCcathepsin L Skin condition diagnosis kit including the above.

[Claim 111]

The method of claim 8,

The skin or skin secretion is a kit for diagnosing skin conditions comprising at least one of keratin and sebum of the skin.

[Claim 12]

The method of claim 8,

A skin or skin secretion collecting kit comprising at least one of a patch, paper, film, and stripping tape.

[Claim 13]

The method of claim 8,

The biomarker measuring unit includes at least one selected from the group consisting of an enzyme, a substrate, and a surfactant,

The enzyme or substrate is a skin condition diagnosis kit which is an enzyme or substrate that specifically reacts with the biomarker to be measured.

[Claim 14]

The method of claim 8,

The biomarker measuring unit includes a kit for diagnosing skin conditions including a fluorescent or color display unit.

[Claim 15]

The method of claim 14,

The fluorescence or color indicator is a skin condition diagnosis kit that displays fluorescence or color depending on the degree of the measured biomarker.

[Claim 16]

The method of claim 14,

The fluorescence or color indicator is a kit for diagnosing skin conditions in which the degree of fluorescence or color display varies depending on the degree of the measured biomarker.

[Claim 17]

The method of claim 8,

The kit further comprises a reader for observing a fluorescence or color change.

[Claim 18] The method according to claim 8 or 17,

The kit includes a skin condition diagnosis key that further includes instructions including instructions for use, skin condition indicators, and one skin care method.