WO2018193565A1 - Method for evaluating impact of test substance or product on skin - Google Patents
Method for evaluating impact of test substance or product on skin Download PDFInfo
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- WO2018193565A1 WO2018193565A1 PCT/JP2017/015796 JP2017015796W WO2018193565A1 WO 2018193565 A1 WO2018193565 A1 WO 2018193565A1 JP 2017015796 W JP2017015796 W JP 2017015796W WO 2018193565 A1 WO2018193565 A1 WO 2018193565A1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
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- 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
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- 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
- G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
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- 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
Definitions
- the present invention relates to a method for evaluating the influence of a test substance or article on skin.
- Skin has the function of preventing the loss of moisture and biological components from the inside of the body and the entry of foreign substances from the outside. Such a function is generally called a “skin barrier function”.
- the barrier function of the skin is mainly borne by the stratum corneum present in the outermost layer of the skin.
- the water content of the stratum corneum is about 20% to 30% on the skin surface.
- the water content gradually increases as it goes in the depth direction, and the water content reaches about 70% in the epidermis.
- stratum corneum is a part that is touched by human eyes, when a foreign substance such as a chemical substance enters the skin, skin troubles such as inflammation are recognized at a glance.
- skin troubles may occur when the skin is covered or brought into contact with absorbent articles such as diapers and sanitary products, wound protection materials, masks, rubber gloves and the like.
- absorbent articles such as diapers and sanitary products, wound protection materials, masks, rubber gloves and the like.
- many women suffer from skin problems such as itching and rash during use of sanitary products. Baby diaper rash is also frequently observed.
- the permeability of a substance to the stratum corneum is one of the very important factors in the medical field and the beauty field.
- Non-Patent Document 1 a method of applying methyl nicotinate to the skin and evaluating it as a method for evaluating the barrier function of the skin is widely used (for example, see Non-Patent Documents 1 to 3). In this method, the barrier function of the skin is evaluated based on erythema induced by the vasodilatory action of methyl nicotinate.
- erythema is also affected by factors other than skin function, such as blood flow, outside air temperature, increased body temperature, decreased body temperature, body movement, sweating, and subject skin color. Therefore, in the above-described method, the evaluation result of the barrier function may be influenced by various factors (excluding the controlled environment and situation). Moreover, since methyl nicotinate has a vasodilatory effect, it may be a burden itself to apply methyl nicotinate to the skin of a subject.
- a test substance or article is brought into contact with human skin, and the amount of change in the moisture content or distribution of the skin before and after contact is compared with a reference value.
- a method for evaluating a test substance or article for evaluating an influence on a function The water content or distribution of the skin, measured by the water load test on the skin with different water loads, is related to the skin barrier function, and the skin water content or water distribution and the skin barrier function Therefore, the present invention relates to a method for evaluating a test substance or article, in which the moisture content or moisture distribution of the skin when a change in the barrier function of the skin is confirmed is set as the reference value.
- the present invention also provides a test substance by measuring the amount of water for each depth of human skin after contacting the test substance or article with human skin, and obtaining the moisture distribution after contact with the test substance or article.
- a test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin by comparing the moisture distribution after contact of the article with a reference moisture distribution, Before applying water or aqueous solution to human skin, measure the amount of water at each depth of human skin to obtain the initial moisture distribution, and immediately after loading water or aqueous solution to human skin, Obtain water distribution after loading water or aqueous solution by measuring the amount of water at each skin depth, apply the indicator substance to the skin, measure the permeability of the indicator substance, and determine the water or aqueous solution from the initial moisture distribution.
- the present invention relates to a test substance or article evaluation method in which the moisture distribution after loading with water or an aqueous solution at each depth is used as the reference moisture distribution.
- the present invention is a method for evaluating the effect of a test substance or article on the barrier function of human skin, Obtain the moisture distribution by measuring the amount of moisture at each skin depth before and after contact of the test substance or article with human skin, and the initial angle before contact in the moisture distribution before and after contact.
- the present invention relates to a test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin using as an index the difference in moisture content deeper than the layer thickness.
- FIG. 2 (a) is a graph showing the moisture distribution when the moisture load on the skin is 0 mL / cm 2
- FIG. 2 (b) shows that the moisture load on the skin is 0.02 mL / cm 2
- 2 (c) is a graph showing the moisture distribution when the moisture load on the skin is 0.04 mL / cm 2
- FIG. 2 (d) shows the moisture distribution on the skin. It is a graph which shows a moisture distribution in case water load amount of is 0.20 mL / cm ⁇ 2 >. It is a graph which shows the moisture distribution in the skin of the depth direction measured using the confocal Raman spectroscopic microscope before and behind sticking of two types of absorbent articles.
- the present invention provides a test substance or article evaluation method that evaluates the influence of a test substance or article on the skin barrier function in a non-invasive manner, with less burden on the subject, safe, simple, and in a short time.
- the swelling state (hydration state) of the stratum corneum located on the outermost layer of the skin affects the barrier function of the skin, and the permeability of the substance to the skin changes accordingly. Therefore, if it is possible to grasp the hydration state of the stratum corneum caused by the test substance or article coming into contact with the skin, the test substance or article has less burden on the subject as to how the test substance or article affects the barrier function of the skin, It is expected to be evaluated without being affected by various factors. However, it has not been elucidated in detail about the relationship between the change in substance permeability to the skin and the increase / decrease of the moisture content of the skin and the moisture distribution in the depth direction of the skin.
- the present inventors examined the relationship between the permeability of a substance in living human skin and the moisture content of the stratum corneum located on the outermost surface of the skin and the moisture distribution in the depth direction. As a result, we found for the first time in living humans that there is a high correlation between the moisture content of the stratum corneum and the moisture distribution in the depth direction and the permeability of substances to the skin. Then, the test substance or article becomes a skin barrier from the change in the skin moisture content or the moisture distribution in the depth direction in a living human measured non-invasively before and after the test substance or article is brought into contact with the human skin. It was found that the effect on the function can be evaluated. The present invention has been completed based on these findings.
- the skin barrier function is evaluated from the moisture content and moisture distribution of the skin before and after contact with the test substance or article based on the relationship between the moisture content and moisture distribution of the skin and the barrier function of the skin. Therefore, according to the present invention, the influence of the test substance or article on the skin can be known safely, simply and in a short time.
- the test substance or article is brought into contact with human skin (S1), the amount of change in the moisture content or moisture distribution of the skin measured before and after contact with the test substance or article, Compare the moisture distribution of the skin with the reference value or reference moisture distribution (S2), and evaluate the effect of the test substance or article on the skin barrier function from the comparison result with the reference value or reference moisture distribution (S3) .
- analysis and evaluation of the influence on the skin barrier function are performed based on the relationship between the amount of moisture in the skin or the distribution of moisture distribution and the barrier function of the skin.
- carrier function refers to the permeability of a substance from the outside to the inside of the skin.
- the “test substance” means a substance (for example, a compound, a chemical substance, a pigment, a secretion or excrement derived from a living body, a moisturizing agent, or the like that penetrates into the skin by contact with the skin or application to the skin. External preparations, cosmetics).
- the “test substance” and the “indicator substance” described later include water-soluble components and hydrophobic components.
- substances include water or water vapor, physiological saline, phosphate buffered saline, methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4 (tartrazine), red No. 215 (rhodamine B stearate), cafe In, ubinal, carotenoids, retinol, urine, sweat, menstrual blood, feces, simulated urine, simulated menstrual blood, simulated feces, and active ingredients contained in skin moisturizers, topical skin preparations, skin cosmetics or steam heating devices Medicinal ingredients are listed.
- the skin moisture content or moisture distribution at the site where the barrier function is evaluated is measured.
- the measurement of the skin moisture content or moisture distribution is preferably performed noninvasively. In this case, the burden on the subject is small, and the skin barrier function can be safely evaluated.
- the method for noninvasively measuring the moisture content or moisture distribution in the stratum corneum can be appropriately selected from conventional methods. Examples thereof include a method using Raman spectroscopy, skin capacitance measurement means, high-frequency current measurement means, infrared spectroscopy, or near infrared spectroscopy.
- an apparatus capable of measuring skin moisture content or moisture distribution in accordance with Raman spectroscopy, skin capacitance measuring means, etc., when the test substance or article is loaded on the skin. Use to measure. Then, the influence of the substance or article on the skin barrier function is evaluated from the amount of change. As shown in Examples described later, when excessive moisture is applied to human skin, the substance permeability is enhanced and the skin barrier function is lowered. In response to such findings, the present inventors evaluated the skin barrier function based on the skin moisture content or the change in the moisture distribution when the test substance or article is loaded on the skin or the reference moisture distribution. I found what I could do for the first time.
- the substance permeability does not increase until the skin moisture content reaches a certain value. However, when the moisture content of the skin increases above a certain value, the substance permeability increases.
- the permeability of methyl nicotinate which is a water-soluble substance, was evaluated as a representative example of the indicator substance. A water-soluble substance is likely to be distributed into the skin as the amount of moisture in the skin increases, and may penetrate into the skin.
- the reference value and the reference moisture distribution for evaluating the influence of the test substance or article on the skin barrier function can be set as appropriate.
- the setting method of the reference value and the reference moisture distribution in the present invention will be described based on specific examples.
- the present invention is not limited to this.
- the water load test is performed on the skin by changing the water load, and the moisture content or moisture distribution of the skin at each water load is measured by a conventional method.
- the measured moisture content or moisture distribution of the skin is associated with the barrier function of the skin.
- the association between the moisture content or moisture distribution of the skin and the barrier function of the skin is defined as “the reference value of the moisture content or moisture distribution of the skin when a change is confirmed in the skin barrier function. ”Is set.
- the method for setting the “reference value” include a method for setting from the permeability of the indicator substance from the outside to the inside of the skin. Following the water load test on the skin described above, the load test of the index substance to the skin was conducted to measure the permeability of the index substance by a conventional method, and the moisture of the skin when the change in the permeability of the index substance was observed The amount or moisture distribution can be a “reference value”. In this embodiment, when a change is confirmed in relation between the amount of moisture loaded and the permeability of the indicator substance, or when the permeability of the indicator substance reaches a certain state, can do.
- the association between the moisture content or moisture distribution of the skin and the barrier function of the skin is the human when the association between the permeability of the indicator substance and the load of water or aqueous solution is changed.
- the water distribution after loading with water or an aqueous solution at each skin depth is set as the “reference water distribution”.
- the amount of water at each depth of the human skin is measured to obtain an initial moisture distribution.
- water or an aqueous solution is loaded onto the human skin for a certain period of time, and immediately after the end of the loading, the moisture content at each depth of the human skin is measured to obtain the moisture distribution after loading with the water or aqueous solution.
- the indicator substance is applied to the skin and the penetration degree of the indicator substance is measured. Then, the measurement is repeated while increasing or decreasing the load of water or aqueous solution until the relationship between the permeability of the indicator substance and the load of water or aqueous solution changes. Based on the measurement results, the water distribution after loading of water or aqueous solution at each depth of human skin when the relationship between the permeability of the indicator substance and the load of water or aqueous solution changes is determined as the reference water content. It can be a distribution.
- the indicator substance for evaluating the permeability to the skin can be appropriately selected.
- the moisture content deeper than the initial stratum corneum thickness is associated with the skin barrier function.
- the moisture distribution is obtained by measuring the amount of moisture at each skin depth before and after the test substance or article is brought into contact with human skin. Then, the moisture distribution before and after contact is compared, and the test substance or article is evaluated using as an index the difference in moisture content deeper than the initial stratum corneum thickness before contact.
- the amount of moisture or the moisture distribution in any region or part from the stratum corneum surface layer to the granular layer may be measured.
- the characteristics of the moisture content or moisture distribution measuring device used in the present invention for example, the intensity of Raman scattering due to the excitation wavelength applied to the inside of the skin, the moisture content or moisture in the region from the skin surface to a predetermined depth You may measure distribution or the moisture content of the site
- the “initial stratum corneum thickness” refers to the thickness of the stratum corneum before moisture, a test substance or an article is loaded.
- the initial stratum corneum thickness from the Raman spectrum can be measured according to the method described in IFSCC Magazine, 2009, vol. 12 (1), p. 9-15. Calculate the moisture content at each depth from the Raman spectrum measured according to the usual method, and create a graph plotting the numerical value of the moisture content approximately calculated for each fixed distance (eg 1 ⁇ m) in the depth direction. Get distribution profile. Then, the obtained moisture profile is approximated by two straight lines, and the depth to the intersection can be defined as the stratum corneum thickness from the stratum corneum surface layer.
- the signal intensity derived from the CH stretching vibration of the protein shows a maximum value on the skin side slightly from the boundary between the window material of the objective lens and the skin, and then goes deeper. Signal strength decreases. This is because, in the skin, the signal is attenuated by the diffusion of light as it goes deeper. Since there is no signal derived from protein CH stretching vibration in the window material, the signal intensity derived from CH stretching vibration is 0 when the focal point of the laser is within the window material. On the other hand, when the signal intensity derived from the CH stretching vibration when the laser is focused in the skin is 100, the signal intensity derived from the CH stretching vibration is 50 when the focus comes to the interface between the window material and the skin.
- the point where the signal intensity derived from the CH stretching vibration shows the maximum half value is defined as the boundary between the skin and the window material.
- the window material may be detected even though the signal intensity derived from the CH stretching vibration is weak. Therefore, when the moisture content is calculated and analyzed according to the method described in JP2012-50739A, the moisture content will be high, and the interface between the skin and the window material, that is, the moisture content on the outermost surface of the skin is not appropriate. It is theoretically clear that there is no data. Further, the spatial resolution in the depth direction of the confocal Raman spectroscopic device used in this condition is 2 ⁇ m, and the water amount for each 1 ⁇ m is interpolated and calculated in the analysis.
- the data analysis when determining the moisture content on the skin surface, the data from the actual skin surface to at least 5 ⁇ m is omitted, and the skin surface is not less than 5 ⁇ m.
- the part of the skin is the “skin surface” in the data analysis, the data of at least 2 ⁇ m from the actual skin surface is omitted, and the part of 2 ⁇ m or more from the skin surface is the “skin surface” in the data analysis More preferably, it is more preferable to omit the data from the actual skin surface to 2 ⁇ m, and to set the portion of 2 ⁇ m from the skin surface as the “skin surface” in the data analysis.
- a method for approximately calculating the amount of water from a Raman spectrum obtained by measurement using a confocal Raman spectroscopic microscope can be performed with reference to the method described in JP 2012-50739 A.
- the initial stratum corneum thickness measured by Raman spectroscopy is usually about 10 ⁇ m to 20 ⁇ m and an average of about 13 ⁇ m. Therefore, in the present invention, the moisture content of the skin is from 2 to 40 ⁇ m deep from the actual skin surface, preferably from 2 to 35 ⁇ m deep from the skin surface, more preferably from the skin surface. It is preferable to measure and analyze up to a depth of 2 to 20 ⁇ m, more preferably a depth of 9 to 15 ⁇ m from the skin surface, more preferably a depth of about 13 ⁇ m from the skin surface.
- the skin moisture amount or moisture distribution measured by Raman spectroscopy, skin capacitance measuring means, or the like has a strong relationship with the skin barrier function. Specifically, when the amount of change in the amount of moisture in the skin increases by a certain value or more due to contact with the test substance or article, the permeability of substances such as methyl nicotinate to the skin is enhanced.
- the test substance or article is obtained from the amount of change in the moisture content or moisture distribution measured noninvasively. Evaluate the effects on skin barrier function.
- the barrier function is reduced (for example, the permeability of the substance from the outside to the inside of the skin) Can be evaluated).
- the amount of moisture in the skin after contact with the test substance or article changes with an amount of change smaller than the reference value, it can be evaluated that the barrier function has not changed.
- the skin capacitance measuring device (calculating the relative value (AU) of moisture calculated from the capacitance) for the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes
- the reference value for the amount of change in the moisture content of the skin before and after contact with the test substance or article is set to 8 (AU), preferably 12 (AU), more preferably 16 (AU). To do.
- the amount of moisture in the skin after contact with the test substance or article actually measured increases more than the reference value, the permeability of the substance from the outside to the inside of the skin is enhanced, and contact with the test substance or article It can be evaluated that the barrier function is lowered.
- the amount of change in the amount of water is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
- the moisture distribution of the skin before and after contact with the test substance or article is compared. Based on the initial stratum corneum thickness, the amount of skin moisture deeper than the initial stratum corneum due to the test substance or article contact When it increases, it becomes easy to enhance the permeability of the substance from the outside to the inside of the skin, and it can be evaluated that the barrier function is lowered.
- the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is the target, from the actual skin surface to a predetermined depth (preferably a depth of 40 ⁇ m from the skin surface).
- the moisture distribution of the skin before and after contact with the test substance or article is measured by Raman spectroscopy. To do. Then, the moisture distribution in the skin before and after contact with the test substance or article is compared, and when the skin moisture content in the deep part is deeper than the initial stratum corneum thickness due to contact with the test substance or article, It can be evaluated that the permeability of the substance is enhanced and the barrier function is lowered by contact with the test substance or article.
- the moisture content on the skin surface before and after contact with the test substance or article can be measured, and the influence on the barrier function can be evaluated from the amount of change in the moisture content on the skin surface due to contact with the test substance or article.
- the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is targeted, and the moisture content of the skin before and after contact with the test substance or article on the skin surface is determined by Raman spectroscopy. Measure and compare the moisture content of the skin before and after contact with the test substance or article.
- the reference value of the amount of change in the moisture content on the skin surface obtained by Raman spectroscopy before and after contact with the test substance or article is 2.0 times, more preferably 2.5 times, and even more preferably 3. Set to 0 times.
- the surface moisture content of the skin after contact with the actually measured test substance or article increases above the reference value, the permeability of the substance from the outside of the skin to the inside increases, and the test substance or article It can be evaluated that the barrier function is lowered by the contact.
- the amount of change in the surface moisture content of the skin is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
- the moisture content at the depth corresponding to the initial stratum corneum thickness before and after contact with the test substance or article is measured, and the change in the moisture content at the depth corresponding to the initial stratum corneum due to the contact with the test substance or article.
- the effect on the barrier function can also be evaluated from the amount. Specifically, the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is targeted, and the skin before and after contact with the test substance or article at the site corresponding to the initial stratum corneum thickness The moisture content is measured by Raman spectroscopy, and the moisture content of the skin before and after contact with the test substance or article is compared.
- the reference value of the amount of change before and after the contact of the test substance or article with the water content at the site corresponding to the initial stratum corneum thickness obtained by Raman spectroscopy is 8%, preferably 11%, more preferably Is set to 14%. Then, when the amount of water at the site corresponding to the initial stratum corneum thickness after contact with the actually measured test substance or article increases more than the reference value, the permeability of the substance from outside the skin to the inside is enhanced. It can be evaluated that the barrier function has decreased due to contact with the test substance or article. On the other hand, when the amount of change in the amount of water at the site corresponding to the initial stratum corneum thickness is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
- the skin inside the forearm of a healthy person after washing and acclimatization for at least 20 minutes is used as a target, and is measured by Raman spectroscopy to a predetermined depth from the skin surface (preferably a depth from the skin surface).
- the reference value of the amount of change in the area under the curve before and after contact of the test substance or article obtained by Raman spectroscopy is 1.4 times, preferably 1.6 times, more preferably 1.8. Set to double. Then, when the area under the curve from the skin surface to the predetermined depth after contact with the actually measured test substance or article increases beyond the reference value, the permeability of the substance from outside the skin to the inside increases. It can be evaluated that the barrier function has decreased due to contact with the test substance or article. On the other hand, when the amount of change in the area under the curve is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
- the “lower area curve” indicates the moisture content in the stratum corneum, and is obtained by integrating the moisture content for each depth in the depth direction.
- Measurement of the moisture content of the skin by Raman spectroscopy can be performed with reference to the method described in JP2012-50739A.
- This method approximates a skin sample with a ternary system of protein, water, and lipid, creates a corrected Raman spectrum by subtracting the contribution from the lipid-derived Raman spectrum from the measured Raman spectrum of the skin, and generates a protein-derived CH
- This is a method of measuring the amount of skin moisture based on the ratio of the signal intensity derived from the OH stretching vibration of water to the stretching signal intensity.
- a method for measuring the moisture content of the skin by Raman spectroscopy will be described in detail. However, the present invention is not limited to this.
- the baseline fluorescence is corrected. Since the Raman spectrum of the measured skin to eliminate the distortion of baseline by fluorescence, from the data of the wavenumber region of 2000 ⁇ 2800 cm -1 and 3800 ⁇ 4200cm -1, a baseline based on cubic function approximation using the least square method Predict and subtract this.
- the corrected skin spectrum excluding the influence of fluorescence is superimposed on four Raman spectra of degreased and dried stratum corneum, model sebum, model intercellular lipid and water using equation (4) described in JP2012-50739A. Approximate together. That is, the coefficients C 1 to C 4 in the equation (4) are determined by the least square method.
- C 2 I SEB ( ⁇ ) is the contribution of sebum
- C 3 I CER ( ⁇ ) is the contribution of the intercellular lipid.
- the contribution amount of the model lipid in the Raman spectrum of the model lipid shown below is estimated by the least square method.
- a corrected Raman spectrum represented by the formula (5) described in JP 2012-50739 A is obtained by subtracting the contribution of sebum and the contribution of intercellular lipids.
- CH stretch signal (wavenumber region: 2800 ⁇ 3030cm -1) in the correction Raman spectra of the formula (5) OH stretching vibration peak area (wavenumber region: 3100 ⁇ 3750cm -1) to the peak area ratio of Is calculated.
- the moisture content (mass%) of the skin is calculated based on a line (for example, see FIG. 5 of JP2012-50739A).
- a horny layer piece is excised with a knife from the heel of a degreased and dried horny layer healthy male (40's), and then immersed in chloroform-methanol (1: 1) overnight. Thereafter, a sample of the stratum corneum is taken out and naturally dried to prepare a degreased and dried stratum corneum from which oil-soluble components such as lipids and water-soluble components such as amino acids have been removed.
- Model sebum The following components are mixed to prepare model sebum.
- Examples of articles used in the present invention include absorbent articles (absorbent cotton, sanitary napkins, paper diapers, cloth diapers, training pants, adult diapers, sweat pads, urine pads, puerperium pads, panty liners, absorbent pads, etc.) , Steam heaters, wound protection materials (such as bandages, gauze, bandages, etc.), masks, and rubber gloves.
- Examples of the test substance include water or water vapor, cosmetics, humectants or external preparations, active ingredients and medicinal ingredients contained in steam heating devices, physiological saline, phosphate buffered saline, and biologically derived secretions. And urine such as urine, sweat, menstrual blood, and feces.
- simulated urine, simulated sweat, simulated menstrual blood, simulated stool, etc. are included in living body-derived secretions and excretion.
- the contact time of the test substance or article with the skin can also be set as appropriate.
- the time for contacting the test substance or article with human skin is 1 hour or more and 6 hours or less, preferably 2 hours or more and 4 hours or less, more preferably 2.5 hours or more and 3.5 hours or less, and further preferably About 3 hours.
- the evaluation method of the present invention measures the skin moisture content or moisture distribution noninvasively, and relates the relationship between the amount of change in skin moisture content or moisture distribution due to contact with an article or test substance and the skin barrier function. Based on the sex, the barrier function of the skin, in particular the permeability of the substance, is evaluated from the measured amount of change in the skin water content.
- the amount of change in skin moisture or moisture distribution measured non-invasively is also described.
- the state of the stratum corneum can be evaluated. For example, the amount of moisture in the skin before and after loading of the test substance or article, measured using a skin capacitance measuring device, for the skin inside the forearm of a healthy subject after washing the skin and acclimatizing for at least 20 minutes
- the reference value of the change amount is set to 8 (AU), preferably 12 (AU), more preferably 16 (AU).
- the barrier function of the stratum corneum is lowered when the moisture content of the skin after loading the actually measured test substance or article increases by more than the reference value.
- the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is the target, and from the skin surface to a predetermined depth (preferably to a depth of 40 ⁇ m from the skin surface, more preferably The water distribution of the skin before and after contact with the test substance or article at a depth of 35 ⁇ m from the skin surface, more preferably to a depth of 20 ⁇ m from the skin surface) is measured by Raman spectroscopy.
- the moisture distribution in the skin before and after contact with the test substance or article is compared, and when the skin moisture content in the deeper part than the initial stratum corneum thickness is increased by contact with the test substance or article, the barrier function of the stratum corneum is It can be evaluated that it has decreased.
- the skin moisture before and after contact with the test substance or article at a depth of 2 ⁇ m from the skin surface was examined for the skin inside the forearm of a healthy person after the skin was washed and acclimatized for at least 20 minutes. Measure by spectroscopy. Then, the surface moisture content of the skin before and after contact with the test substance or article is compared.
- the reference value of the amount of change of the obtained skin surface moisture before and after contact with the test substance or article is set to 2.0 times, more preferably 2.5 times, and even more preferably 3.0 times. And it can be evaluated that the barrier function of the stratum corneum is reduced when the surface moisture content of the skin after contact with the test substance or article actually measured increases above the reference value.
- the amount of moisture in the skin before and after contact with the test substance or article at the site corresponding to the initial stratum corneum thickness is measured for the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes. Measured by Raman spectroscopy. Then, the moisture content of the skin before and after contact with the test substance or article is compared.
- the reference value of the amount of change before and after the contact of the test substance or article with the moisture content at the site corresponding to the initial stratum corneum thickness obtained by Raman spectroscopy is 8%, preferably 11%, more preferably 14%. Set. And when the moisture content of the site
- the skin inside the forearm of a healthy person after washing and acclimatization for at least 20 minutes is used as a target, and is measured by Raman spectroscopy to a predetermined depth from the skin surface (preferably a depth from the skin surface).
- the area under the curve of water profile is determined at a depth of 15 ⁇ m, more preferably a depth of about 13 ⁇ m from the skin surface.
- the reference value of the amount of change in the area under the curve of the moisture content of the skin after contact with the test substance or article for a certain period of time relative to the area under the curve of the moisture content of the skin before contact with the test substance or article is preferably 1.4 times, preferably It is set to 1.6 times, more preferably 1.8 times.
- the barrier function of the stratum corneum is reduced. Can be evaluated.
- Absorbent articles absorbent cotton, sanitary napkins, disposable diapers, cloth diapers, training pants, adult diapers, sweat pads, urine pads, puerperium pads, panty liners, absorbent pads, etc.
- products such as skin cosmetics or external preparations, wound protection materials (such as bandages, gauze, bandages, etc.), masks or rubber gloves, and steam heaters.
- the performance of these products e.g., by evaluating the barrier function of the skin before and after wearing using the evaluation method of the present invention
- the ability to absorb moisture (wet), the ability of water vapor (steaming) to spread to the outside, and the likelihood of skin problems) can be evaluated safely and simply in a short time.
- an absorbent article containing physiological saline, simulated urine, or the like may be used for the evaluation. If the moisture content or moisture distribution of the skin after the wearing or application of the article is evaluated according to the present invention, the permeability of the substance is evaluated without the need to apply a substance such as methyl nicotinate used in this example to human skin.
- the substance permeability can be estimated from the moisture content or moisture distribution of the skin before and after wearing when comparing the current product with the improved product. it can. If it does in this way, development of an absorptive article etc. can be performed efficiently.
- active ingredients contained in cosmetics, external preparations and the like can be permeated into the skin using the hydration state of the skin (for example, J. Invest. Dermatol., 1963, vol. 41, p. 307-311).
- the active ingredient contained in these products can be evaluated by using the evaluation method of the present invention to evaluate the barrier function of the skin before and after application, application, or wearing on the skin of cosmetics, external preparations, steam heaters, etc.
- the permeability to the skin such as can be evaluated safely and simply in a short time.
- the system for evaluating the influence of the test substance or article on the skin barrier function relates to the relationship between the amount of change in the moisture content or distribution of the skin due to the contact of the test substance or article and the skin barrier function. Information on sex is stored, and the influence is evaluated from the amount of change in the moisture content or moisture distribution of the skin due to contact with the test substance or article based on the information on the relevance.
- the system of the present invention is a so-called computer and includes, for example, a CPU (Central Processing Unit), a memory, an input / output interface, and the like that are connected to each other via a bus.
- a CPU Central Processing Unit
- the memory is a RAM (Random Access Memory), a ROM (Read Only Memory), a hard disk, a portable storage medium, or the like.
- the input / output interface is connected to a user interface device such as a display device or an input device.
- the input / output interface may be connected to a communication device or the like that communicates with another computer via a network.
- the evaluation method and the evaluation system of the present invention target human skin, preferably human (healthy person) skin having a normal barrier function in a state in which no test substance or article is loaded.
- the present invention further discloses the following evaluation method and evaluation system.
- a test substance or article is brought into contact with human skin, Compare the amount of moisture or moisture distribution in the skin before and after contact with the reference value, From the comparison results, the effect of the test substance or article on the skin barrier function is evaluated.
- a method for evaluating a test substance or article The water content or distribution of the skin, measured by a water load test on the skin with different water loads, is associated with the barrier function of the skin, From the relationship between the moisture content or moisture distribution of the skin and the barrier function of the skin, the moisture content or moisture distribution of the skin when the change is confirmed in the skin barrier function is set as the reference value. Evaluation method of test substance or article.
- ⁇ 2> In order to confirm the change in the barrier function of the skin, the permeability of the indicator substance from the outside to the inside of the skin is further measured, and the reference value is set from the result of the permeability of the indicator substance.
- ⁇ 3> When the amount of skin moisture after contact with the test substance or article increases with a change amount greater than or equal to a reference value, the barrier function is reduced, preferably the permeability of the substance from the outside of the skin to the inside is increased, The method according to ⁇ 1> or ⁇ 2>, wherein ⁇ 4> When the amount of change in skin moisture after contact with the test substance or article is smaller than the reference value, the barrier function is not changed, preferably the permeability of the substance from the outside to the inside is enhanced. The method according to any one of ⁇ 1> to ⁇ 3>, wherein the method is evaluated as having no.
- ⁇ 5> Measure the moisture content for each depth of human skin after contacting the test substance or article with human skin, and obtain the moisture distribution after contact of the test substance or article, A test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin by comparing the moisture distribution after contact of the test substance or article with a reference moisture distribution.
- the indicator Measure the permeability of the substance, From the initial moisture distribution to the moisture distribution immediately after the end of the load of water or aqueous solution, and the permeability of the indicator substance, determine the relationship between the permeability of the indicator substance and the load of water or aqueous solution, The water distribution after loading with water or an aqueous solution at every depth of the human skin when the relevance is changed as the reference water distribution, Evaluation method of test substance or article.
- ⁇ 6> Measure the moisture content at a predetermined depth position of the human skin before and after contact with the test substance or article, and obtain the difference between the moisture content before contact and the moisture content after contact; The evaluation method according to ⁇ 5>, wherein the influence on the skin barrier function is evaluated based on the obtained difference in water amount and the reference water distribution.
- the indicator substance is methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4, red No. 215, caffeine, ubinal, carotenoid, retinol, and skin moisturizer, skin external preparation, skin cosmetic, or steam
- ⁇ 8> The method according to any one of ⁇ 1> to ⁇ 7>, wherein the moisture content of the skin is measured and analyzed by a skin capacitance measuring unit.
- ⁇ 9> Skin of healthy subject is evaluated, skin moisture after contact with test substance or article is measured by skin capacitance measuring means, and skin moisture after contact with test substance or article is tested.
- the barrier function decreases.
- ⁇ 11> Up to a depth of 2 to 40 ⁇ m from the skin surface, preferably up to a depth of 2 to 35 ⁇ m from the skin surface, more preferably up to a depth of 2 to 20 ⁇ m from the skin surface, more preferably The water content or water distribution up to a depth of 9 to 15 ⁇ m from the skin surface, more preferably a depth of about 13 ⁇ m from the skin surface is measured and analyzed by Raman spectroscopy, ⁇ 1> to ⁇ 7. The method according to any one of 7>.
- the skin of a healthy person is the subject of evaluation, up to a predetermined depth from the skin surface, preferably up to a depth of 40 ⁇ m from the skin surface, more preferably up to a depth of 35 ⁇ m from the skin surface, more preferably
- the skin moisture distribution before and after contact with the test substance or article up to a depth of 20 ⁇ m from the skin surface is measured by Raman spectroscopy, and the skin moisture distribution before and after contact with the test substance or article is compared.
- the barrier function is lowered, preferably the permeability of the substance from the outside to the inside of the skin is increased.
- Skin surface moisture after contact with a test substance or article obtained by Raman spectroscopy is 2 with respect to the skin surface moisture before contact with the test substance or article.
- the barrier function is lowered, preferably the permeability of the substance from the outside of the skin to the inside is increased,
- the skin surface moisture after contact of the test substance or article obtained by Raman spectroscopy is 3 with respect to the skin surface moisture before contact of the test substance or article.
- the barrier function is not changed, preferably the permeability of the substance from outside the skin to the inside is The method according to ⁇ 11>, wherein the method is evaluated as not enhanced.
- the skin of a healthy person is the subject of evaluation, and the water content of the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the value before contact with the test substance or article.
- the barrier function is lowered, preferably from the outside of the skin to the inside.
- the method according to ⁇ 11> wherein the permeability of the substance is evaluated to be enhanced.
- the moisture content of the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the value before contact with the test substance or article. If the amount of water in the region corresponding to the initial stratum corneum thickness is less than 14% increase, preferably less than 11% increase, more preferably less than 8% increase, the barrier function is not changed, Preferably, the method according to ⁇ 11> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is not enhanced.
- the skin of a healthy person is evaluated, and is obtained by Raman spectroscopy up to a predetermined depth from the skin surface, preferably a depth of 2 to 40 ⁇ m from the skin surface, more preferably from the skin surface.
- a depth of 2 to 35 ⁇ m more to a depth of 2 to 20 ⁇ m from the skin surface, more preferably to a depth of 9 to 15 ⁇ m from the skin surface, more preferably to a depth of 13 ⁇ m from the skin surface.
- the area under the curve of the moisture content of the skin after contact with the test substance or article up to a certain depth is obtained, and the area under the curve of the skin moisture amount after contact with the test substance or article is the contact of the test substance or article
- the barrier function is lowered, preferably skin From outside to inside
- the skin of a healthy person is evaluated, and obtained by Raman spectroscopy, to a predetermined depth from the skin surface, preferably to a depth of 2 to 40 ⁇ m from the skin surface, more preferably from the skin surface.
- a depth of 2 to 35 ⁇ m more to a depth of 2 to 20 ⁇ m from the skin surface, more preferably to a depth of 9 to 15 ⁇ m from the skin surface, more preferably to a depth of 13 ⁇ m from the skin surface.
- the area under the curve of the moisture content of the skin after contact with the test substance or article up to a certain depth is obtained, and the area under the curve of the skin moisture amount after contact with the test substance or article is the contact of the test substance or article If the area under the curve of water content of the previous skin is less than 1.8 times, preferably less than 1.6 times, more preferably less than 1.4 times, the barrier function is not changed, Like Alternatively, the method according to ⁇ 11> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is not enhanced.
- a method for evaluating the influence of a test substance or article on the barrier function of human skin Obtain water distribution by measuring the amount of water at each skin depth before and after contact of the test substance or article with human skin, Evaluate the effect of the test substance or article on the barrier function of human skin, using the difference in water content deeper than the initial stratum corneum thickness before contact in the moisture distribution before and after contact, Evaluation method of test substance or article.
- ⁇ 20> to a depth of 2 to 40 ⁇ m from the skin surface, preferably to a depth of 2 to 35 ⁇ m from the skin surface, more preferably to a depth of 2 to 20 ⁇ m from the skin surface, more preferably The method according to ⁇ 19>, wherein the moisture content is measured and analyzed by Raman spectroscopy up to a depth of 9 to 15 ⁇ m from the skin surface, more preferably a depth of about 13 ⁇ m from the skin surface.
- ⁇ 21> Skin of a healthy subject is evaluated by measuring the moisture distribution of the skin before and after contact with the test substance or article from the skin surface to a predetermined depth by Raman spectroscopy, and before and after contact with the test substance or article
- the barrier function decreases, preferably the permeability of the substance from the outside to the inside of the skin
- ⁇ 22> The method according to any one of ⁇ 1> to ⁇ 21>, wherein the skin moisture content or moisture distribution is noninvasively measured.
- ⁇ 23> The method according to any one of ⁇ 1> to ⁇ 22>, wherein the evaluation is performed on the skin of a human (healthy person) having a normal barrier function in a state in which no test substance or article is loaded.
- the moisture content of the skin is measured according to at least one selected from the group consisting of Raman spectroscopy, skin capacitance measurement means, high-frequency current measurement means, infrared spectroscopy, and near-infrared spectroscopy, The method according to any one of 1> to ⁇ 7>, ⁇ 19>, ⁇ 22> and ⁇ 23>.
- the test substance is water or water vapor, skin cosmetics, skin moisturizer or external preparation for skin, active ingredient or medicinal ingredient contained in steam heating device, physiological saline, phosphate buffered saline, and biological origin
- the method according to any one of ⁇ 1> to ⁇ 24>, wherein the method is at least one selected from the group consisting of secretions and excrements of ⁇ 26>
- the article is an absorbent article (preferably absorbent cotton, a sanitary napkin, a paper diaper, a cloth diaper, a training pant, an adult diaper, a sweat pad, a urine pad, a postpartum pad, a panty liner, or an absorbent pad),
- the method according to any one of ⁇ 1> to ⁇ 24>, wherein the method is a steam heating device, a wound protection material (preferably a bandage, a gauze, or a bandage), a mask, or a rubber glove.
- Time for contacting the test substance or article with human skin is 1 hour or more and 6 hours or less, preferably 2 hours or more and 4 hours or less, more preferably 2.5 hours or more and 3.5 hours or less, and further preferably The method according to any one of ⁇ 1> to ⁇ 27>, wherein is about 3 hours.
- ⁇ 29> Stores information on the relationship between the amount of change in the moisture content or distribution of skin due to contact of the test substance or article with human skin, and the barrier function of the skin.
- Test Example 1 Measurement of change in permeability of methyl nicotinate and moisture content of skin before and after water load
- Measurement of erythema score Healthy males and 10 females (20s to 40s) were subjects. A different amount of water was loaded on the inner side of the forearm, and the permeability of methyl nicotinate after the water load was evaluated. The measurement was carried out after acclimatization for 20 minutes in an environment variable room at 24 to 25 ° C. and a relative humidity of 40 to 50%.
- Moisture load per cm 2 of absorbent cotton is 0 mL (0 mL / cm 2 ), 0.02 mL (0.02 mL / cm 2 ), 0 on 2.5 cm ⁇ 2.5 cm member cut cotton (manufactured by White Cross)
- Add distilled water to make 0.04 mL (0.04 mL / cm 2 ), 0.20 mL (0.20 mL / cm 2 ), place this on the inner side of the forearm, and use a moisture-permeable adhesive bandage (trade name: Skinnagate) to surround it. Fixed and affixed for 3 hours.
- the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm TM Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
- a dressing material trade name: Tegaderm TM Transparent Dressing
- a filter paper (trade name: Advantec No. 2) hollowed out to a diameter of 5 mm containing 5.0 ⁇ L of 0.007 mass% methyl nicotinate aqueous solution was loaded on the test site for 15 seconds. After 30 minutes from the load of methyl nicotinate, the resulting erythema was visually determined according to the following evaluation criteria. From the erythema score obtained by visual determination, the permeability of methyl nicotinate at each water load was evaluated. IBM SPSS version 23 software was used for statistical analysis. The results are shown in Table 1. (Evaluation criteria) Score 0: No response. Score 0.5: Slightly red.
- Score 1 More than half of the area of the filter paper is red.
- Score 2 The area on which the filter paper is applied is clearly red.
- Score 3 larger than the area of the filter paper.
- Score 4 Red area is larger than the area of the filter paper and swollen.
- the load amount of water as compared with the case of 0.00mL / cm 2 or 0.02 mL / cm 2, to load a moisture 0.04 mL / cm 2 or 0.20 mL / cm 2
- the erythema score due to methyl nicotinate was significantly increased. That is, the permeation of methyl nicotinate into the skin was significantly enhanced by applying a certain amount or more of moisture to the skin.
- the skin barrier function is lowered when the water load is at least 0.04 mL / cm 2 . Therefore, the state of the skin when the moisture load is 0.04 mL / cm 2 can be set as a reference state for evaluating the barrier function of the skin.
- Erythema is observed on the skin when the loaded methyl nicotinate penetrates or passes through the skin and acts on the peripheral blood vessels to exert a vasodilating action. As shown in Table 1, erythema is hardly observed on the skin before water load (water load: 0 mL / cm 2 ) even when methyl nicotinate is loaded. This is probably because methyl nicotinate is difficult to penetrate into the skin and cannot reach the peripheral blood vessels. That is, it can be inferred that the barrier function is maintained in the skin before moisture loading. On the other hand, erythema is observed in the skin after a certain amount of water load due to the load of methyl nicotinate. From this result, it can be determined that loading with moisture caused methyl nicotinate to penetrate or pass through the skin, that is, the barrier function of the skin was reduced.
- a moisture load per cm 2 of absorbent cotton is 0 mL (0 mL / 0) on a 2.5 cm ⁇ 2.5 cm member cut cotton (manufactured by White Cross).
- cm 2 0.01mL (0.01mL / cm 2), 0.02mL (0.02mL / cm 2), 0.03mL (0.03mL / cm 2), 0.04mL (0.04mL / cm 2 ), 0.06 mL (0.06 mL / cm 2 ), 0.10 mL (0.10 mL / cm 2 ), and 0.20 mL (0.20 mL / cm 2 ) in distilled water,
- the circumference was fixed with a moisture-permeable adhesive plaster (trade name: Skinnagate) on the inner side of the subject's forearm and applied for 3 hours.
- the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm TM Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
- a dressing material trade name: Tegaderm TM Transparent Dressing
- Skin capacitance measuring device (trade name: Corneometer CM-825, Courage +) before and after removing the member cut cotton, and measuring the moisture content of the skin where the member cut cotton is applied. Measured by Khazaka Germany). Then, a value obtained by subtracting the value before application of the member cut cotton from the value immediately after the removal of the member cut cotton was calculated, and the amount of change in the stratum corneum moisture content was measured. IBM SPSS version 23 software was used for statistical analysis. The results are shown in Table 2.
- the load amount of water of at least 0.04 mL / cm 2 when the load amount of water of at least 0.04 mL / cm 2 can be determined that the skin barrier function is impaired, the state of the skin when the water loading amount of 0.04 mL / cm 2 A reference state for evaluating the skin barrier function was used. Therefore, the amount of change in the amount of skin moisture in this reference state can be used as the reference value in the evaluation method of the present invention.
- the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm TM Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
- a dressing material trade name: Tegaderm TM Transparent Dressing
- a method of approximately calculating the amount of water from the Raman spectrum obtained by measurement using a confocal Raman spectroscopic microscope was performed by referring to the method described in JP2012-50739A. Furthermore, according to the method described in IFSCC Magazine, 2009, vol. 12 (1), p. 9-15, the moisture profile obtained by Raman spectroscopy is approximated by two linear lines, The depth was defined as the initial stratum corneum thickness (the stratum corneum thickness prior to the application of the member cut cotton).
- FIGS. 2 (a) to (d) The results are shown in FIGS. 2 (a) to (d).
- the X-axis indicates the skin depth ( ⁇ m)
- the Y-axis indicates the water content (%).
- the average thickness of the stratum corneum before the water load determined from the graph shown in FIG. 2A was 13 ⁇ m.
- FIG. 2 (b) when the moisture load is 0.02 mL / cm 2 , the skin has an area up to 13 ⁇ m corresponding to the stratum corneum thickness before moisture load compared to before moisture load.
- the amount of change ( ⁇ surface water content) of the surface water content was calculated.
- a signal derived from CH stretching vibration of protein contained in the skin was used as a guide. Details will be described below.
- the signal intensity derived from the CH stretching vibration of the protein of the Raman spectrum shows a maximum value on the skin side slightly from the boundary between the window material of the objective lens and the skin, and then the signal intensity decreases as it goes deeper.
- the signal intensity derived from CH stretching vibration is 0 when the focal point of the laser is within the window material.
- the signal intensity derived from the CH stretching vibration when the laser is focused in the skin is 100
- the signal intensity derived from the CH stretching vibration is 50 when the focus comes to the interface between the window material and the skin.
- the window material may be detected even though the signal intensity derived from the CH stretching vibration is weak. Therefore, in consideration of the spatial resolution of the nano finder, which is the confocal Raman spectroscopic apparatus used for the measurement in the present invention, the water content at a portion 2 ⁇ m deep from the surface, which is the actual measurement value, is defined as “surface water content”. .
- the results are shown in Table 3.
- the surface moisture content before and after the water load changed. Specifically, when the water load was 0.02 mL / cm 2 , the surface water content (%) after the water load increased 1.39 times compared to that before the water load. Similarly, in the case where the moisture load amount is 0.04 mL / cm 2 in the above-described reference state, the surface moisture amount (%) after the moisture load increases 2.40 times compared to before the moisture load, When the water load was 0.20 mL / cm 2 , it increased 2.99 times. Therefore, the moisture content on the skin surface can also be used as the reference value in the evaluation method of the present invention.
- the amount of water at a depth of 13 ⁇ m corresponding to the thickness of the stratum corneum before the water load changed according to the amount of water to be loaded.
- the water load increases after the water load of 0.04 mL / cm 2 where the substance permeability is increased, that is, the water at the depth corresponding to the initial stratum corneum thickness. It was found that the amount increased. Therefore, as the reference value in the evaluation method of the present invention, it is also possible to use the amount of change in the water content at the depth corresponding to the initial stratum corneum thickness.
- the area under the curve of the water content at a depth of 1 ⁇ m to 13 ⁇ m from the skin surface before and after the water load changed. Specifically, when the water load was 0.02 mL / cm 2 , the area under the curve of the water amount after the water load increased 1.20 times compared with that before the water load. Similarly, in the case where the moisture load amount is 0.04 mL / cm 2 in the above-described reference state, the area under the curve of the moisture amount after the moisture load increases 1.59 times compared to before the moisture load, When the water load was 0.20 mL / cm 2 , it increased 1.83 times. Therefore, the area under the curve of the moisture content of the skin up to a predetermined depth from the skin surface can also be used as the reference value in the evaluation method of the present invention.
- the permeability of the substance increases when the water load increases beyond a certain value. That is, the permeability of the substance, that is, the barrier function of the skin, is compared with the skin moisture content or moisture distribution in the reference state from the skin moisture content or moisture distribution measured non-invasively by a corneometer or Raman spectroscopy. Can be evaluated.
- Test Example 2 Performance Comparison Test of Absorbent Article (1) Moisture load on skin One healthy woman (30's) was the subject, and the effect of the absorbent article on the skin on the inner side of the forearm was evaluated. The measurement was performed after acclimatization for 20 to 30 minutes in an environment variable room at 24 to 25 ° C. and a relative humidity of 40 to 50%.
- the amount of skin moisture was increased when the absorbent article B was affixed.
- the absorbent article B is affixed, the amount of water deeper than the initial stratum corneum thickness before the moisture load is increased as compared with the case where the absorbent article A is affixed. Therefore, the barrier function does not change even if the absorbent article A is pasted from the difference in the moisture content deeper than the initial stratum corneum thickness using the moisture distribution, whereas the barrier function when the absorbent article B is pasted Can be evaluated as decreasing.
- the surface moisture content (%) before and after the water load increased 1.19 times after application of the absorbent article A compared to before application. On the other hand, it increased 2.86 times after sticking of the absorbent article B compared with before sticking. Further, the surface moisture content when the absorbent article A was applied was the reference value in Test Example 1, and the increase in the surface moisture content when the moisture load was 0.04 mL / cm 2 (2.40 times). Was smaller than. Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the increase in the amount of surface moisture when the absorbent article B is affixed is greater than the reference value, it can be evaluated that the barrier function decreases when the absorptive article B is affixed.
- the absorbent article B when the absorbent article B was affixed, the water content at a depth of 13 ⁇ m was greatly increased as compared to the case where the absorbent article A was affixed.
- the amount of change in the water content at the depth corresponding to the initial stratum corneum thickness is the reference value in Test Example 1, and the water load is 0.04 mL / cm 2 It was less than the amount of change in water content at the depth corresponding to the initial stratum corneum thickness (9.21 ⁇ 9.2 (%)). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed.
- the barrier function decreases when the absorptive article B is affixed. it can.
- the area under the curve of the water content at a depth of 1 ⁇ m to 13 ⁇ m from the skin surface before and after water load is 1.08 after the application of the absorbent article A and before the application. Doubled.
- the increase was 1.89 times as compared with before application.
- the amount of change in the area under the curve when the absorbent article A was applied was the reference value in Test Example 1, and the increase in the area under the curve when the moisture load was 0.04 mL / cm 2 (1. 59 times). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed.
- the increase in the area under the curve when the absorbent article B is pasted is larger than the reference value, it can be evaluated that the barrier function is lowered when the absorbent article B is pasted.
- the absorbent article A When the absorbent article A was applied and the permeability of the water-soluble substance was actually judged, the permeability of the water-soluble substance from the outside to the inside of the skin was low. That is, from the above results, it can be said that the absorbent article A is an absorbent article that does not increase the permeability of the water-soluble substance to the skin and hardly causes skin troubles.
- the absorbent article B When the absorbent article B was affixed and the permeability of the water-soluble substance was actually judged, it was surprisingly found that the permeability of the water-soluble substance from the outside to the inside of the skin was increased. That is, it can be said that the absorbent article B is an absorbent article in which the permeability of the water-soluble substance to the skin is enhanced and skin trouble may occur.
- test substance or article is brought into contact with human skin according to the method of the present invention, and the effect of the test substance or article on the skin barrier function is determined from the result of measuring the moisture content or moisture distribution of the skin before and after contact. It was confirmed that it is effective as a method for evaluating
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Abstract
A test substance or product evaluation method which involves contacting human skin with a test substance or product, comparing the amount of change in the skin water content or water distribution before and after contact with a reference value, and on the basis of the comparison results, evaluating the impact which the test substance or product has on the skin barrier function, wherein the skin barrier function is associated with the skin water content or water distribution measured by subjecting the skin to a water loading test by changing the water loading amount, and the skin water content or water distribution when a change in the skin barrier function is confirmed is set as the reference value on the basis of the association between the skin barrier function and the skin water content or water distribution.
Description
本発明は、被験物質又は物品が皮膚に与える影響を評価する方法に関する。
The present invention relates to a method for evaluating the influence of a test substance or article on skin.
皮膚は、体内からの水分や生体成分の損失、並びに外部からの異物の進入を防ぐ機能を有している。このような機能は一般に、「皮膚のバリア機能」と呼ばれている。皮膚のバリア機能は主に、皮膚の最外層に存在する角層が担っている。
Skin has the function of preventing the loss of moisture and biological components from the inside of the body and the entry of foreign substances from the outside. Such a function is generally called a “skin barrier function”. The barrier function of the skin is mainly borne by the stratum corneum present in the outermost layer of the skin.
健常な皮膚の一般的性質として、角層の水分量は皮膚表面で約20%~30%であることが知られている。深さ方向へ進むにつれ徐々に水分量は増加し、表皮では水分量は約70%となる。そして、角層は人の目に触れる部分であるため、化学物質などの異物が皮膚に侵入した場合、炎症などの肌トラブルが一目瞭然で認識される。
また、おむつ、生理用品などの吸収性物品や、創傷保護材、マスク、ゴム手袋などで皮膚を被覆又は接触状態にすると、皮膚トラブルが生じる場合がある。実際に多くの女性は、生理用品の使用期間中に、かゆみやかぶれ等の皮膚トラブルに悩んでいる。また、赤ちゃんのおむつかぶれも頻度高く認められている。
これとは逆に、医薬品、医薬部外品、化粧料を皮膚に塗布してケアする場合、これらに含まれる有効成分を角層に十分に浸透させることが重要である。
このように、角層に対する物質の浸透性は、医療分野や美容分野において非常に重要な要素の1つである。 As a general property of healthy skin, it is known that the water content of the stratum corneum is about 20% to 30% on the skin surface. The water content gradually increases as it goes in the depth direction, and the water content reaches about 70% in the epidermis. Since the stratum corneum is a part that is touched by human eyes, when a foreign substance such as a chemical substance enters the skin, skin troubles such as inflammation are recognized at a glance.
In addition, skin troubles may occur when the skin is covered or brought into contact with absorbent articles such as diapers and sanitary products, wound protection materials, masks, rubber gloves and the like. In fact, many women suffer from skin problems such as itching and rash during use of sanitary products. Baby diaper rash is also frequently observed.
On the other hand, when a pharmaceutical, quasi-drug, or cosmetic is applied to the skin for care, it is important that the active ingredient contained therein is sufficiently penetrated into the stratum corneum.
Thus, the permeability of a substance to the stratum corneum is one of the very important factors in the medical field and the beauty field.
また、おむつ、生理用品などの吸収性物品や、創傷保護材、マスク、ゴム手袋などで皮膚を被覆又は接触状態にすると、皮膚トラブルが生じる場合がある。実際に多くの女性は、生理用品の使用期間中に、かゆみやかぶれ等の皮膚トラブルに悩んでいる。また、赤ちゃんのおむつかぶれも頻度高く認められている。
これとは逆に、医薬品、医薬部外品、化粧料を皮膚に塗布してケアする場合、これらに含まれる有効成分を角層に十分に浸透させることが重要である。
このように、角層に対する物質の浸透性は、医療分野や美容分野において非常に重要な要素の1つである。 As a general property of healthy skin, it is known that the water content of the stratum corneum is about 20% to 30% on the skin surface. The water content gradually increases as it goes in the depth direction, and the water content reaches about 70% in the epidermis. Since the stratum corneum is a part that is touched by human eyes, when a foreign substance such as a chemical substance enters the skin, skin troubles such as inflammation are recognized at a glance.
In addition, skin troubles may occur when the skin is covered or brought into contact with absorbent articles such as diapers and sanitary products, wound protection materials, masks, rubber gloves and the like. In fact, many women suffer from skin problems such as itching and rash during use of sanitary products. Baby diaper rash is also frequently observed.
On the other hand, when a pharmaceutical, quasi-drug, or cosmetic is applied to the skin for care, it is important that the active ingredient contained therein is sufficiently penetrated into the stratum corneum.
Thus, the permeability of a substance to the stratum corneum is one of the very important factors in the medical field and the beauty field.
これまでに、ヒトの前腕内側に過剰な水分を含むガーゼやおむつを貼付した場合、皮膚の水分量が増加して角層が膨潤し、その結果ニコチン酸メチルの物質透過性が亢進することが知られている(非特許文献1参照)。このような現象に基づき、皮膚のバリア機能の評価方法として、ニコチン酸メチルを皮膚に塗布し評価する方法が汎用されている(例えば、非特許文献1~3参照)。この方法では、ニコチン酸メチルが有する血管拡張作用により誘導される紅斑に基づき、皮膚のバリア機能を評価する。
しかし紅斑は、血流、外気温度、体温上昇、体温降下、身体の動き、発汗、被験者の皮膚色など、皮膚機能以外の要因によっても影響を受ける。よって、前述の方法ではバリア機能の評価結果が様々な要因(制御された環境や状況を除く)に左右されるおそれがあった。また、ニコチン酸メチルは血管拡張作用を有するため、被験者の皮膚にニコチン酸メチルを塗布すること自体、負担である可能性も考えられる。 So far, when gauze or diapers containing excessive moisture have been applied to the inner side of the human forearm, the moisture content of the skin increases and the stratum corneum swells, resulting in increased substance permeability of methyl nicotinate. It is known (see Non-Patent Document 1). Based on such a phenomenon, a method of applying methyl nicotinate to the skin and evaluating it as a method for evaluating the barrier function of the skin is widely used (for example, see Non-Patent Documents 1 to 3). In this method, the barrier function of the skin is evaluated based on erythema induced by the vasodilatory action of methyl nicotinate.
However, erythema is also affected by factors other than skin function, such as blood flow, outside air temperature, increased body temperature, decreased body temperature, body movement, sweating, and subject skin color. Therefore, in the above-described method, the evaluation result of the barrier function may be influenced by various factors (excluding the controlled environment and situation). Moreover, since methyl nicotinate has a vasodilatory effect, it may be a burden itself to apply methyl nicotinate to the skin of a subject.
しかし紅斑は、血流、外気温度、体温上昇、体温降下、身体の動き、発汗、被験者の皮膚色など、皮膚機能以外の要因によっても影響を受ける。よって、前述の方法ではバリア機能の評価結果が様々な要因(制御された環境や状況を除く)に左右されるおそれがあった。また、ニコチン酸メチルは血管拡張作用を有するため、被験者の皮膚にニコチン酸メチルを塗布すること自体、負担である可能性も考えられる。 So far, when gauze or diapers containing excessive moisture have been applied to the inner side of the human forearm, the moisture content of the skin increases and the stratum corneum swells, resulting in increased substance permeability of methyl nicotinate. It is known (see Non-Patent Document 1). Based on such a phenomenon, a method of applying methyl nicotinate to the skin and evaluating it as a method for evaluating the barrier function of the skin is widely used (for example, see Non-Patent Documents 1 to 3). In this method, the barrier function of the skin is evaluated based on erythema induced by the vasodilatory action of methyl nicotinate.
However, erythema is also affected by factors other than skin function, such as blood flow, outside air temperature, increased body temperature, decreased body temperature, body movement, sweating, and subject skin color. Therefore, in the above-described method, the evaluation result of the barrier function may be influenced by various factors (excluding the controlled environment and situation). Moreover, since methyl nicotinate has a vasodilatory effect, it may be a burden itself to apply methyl nicotinate to the skin of a subject.
本発明は、被験物質又は物品をヒトの皮膚に接触させ、接触前後の皮膚の水分量又は水分分布の変化量を、基準値と比較し、比較結果から、前記被験物質又は物品が、皮膚バリア機能に与える影響を評価する、被験物質又は物品の評価方法であって、
水の負荷量を変えての皮膚への水負荷試験により測定された、皮膚の水分量又は水分分布と、皮膚のバリア機能とが関連付けられ、皮膚の水分量又は水分分布と皮膚のバリア機能との関連性から、皮膚のバリア機能に変化が確認されるときの皮膚の水分量又は水分分布が、前記基準値として設定されたものである、被験物質又は物品の評価方法に関する。 In the present invention, a test substance or article is brought into contact with human skin, and the amount of change in the moisture content or distribution of the skin before and after contact is compared with a reference value. A method for evaluating a test substance or article for evaluating an influence on a function,
The water content or distribution of the skin, measured by the water load test on the skin with different water loads, is related to the skin barrier function, and the skin water content or water distribution and the skin barrier function Therefore, the present invention relates to a method for evaluating a test substance or article, in which the moisture content or moisture distribution of the skin when a change in the barrier function of the skin is confirmed is set as the reference value.
水の負荷量を変えての皮膚への水負荷試験により測定された、皮膚の水分量又は水分分布と、皮膚のバリア機能とが関連付けられ、皮膚の水分量又は水分分布と皮膚のバリア機能との関連性から、皮膚のバリア機能に変化が確認されるときの皮膚の水分量又は水分分布が、前記基準値として設定されたものである、被験物質又は物品の評価方法に関する。 In the present invention, a test substance or article is brought into contact with human skin, and the amount of change in the moisture content or distribution of the skin before and after contact is compared with a reference value. A method for evaluating a test substance or article for evaluating an influence on a function,
The water content or distribution of the skin, measured by the water load test on the skin with different water loads, is related to the skin barrier function, and the skin water content or water distribution and the skin barrier function Therefore, the present invention relates to a method for evaluating a test substance or article, in which the moisture content or moisture distribution of the skin when a change in the barrier function of the skin is confirmed is set as the reference value.
また本発明は、被験物質又は物品をヒトの皮膚に接触させた後のヒトの皮膚の深さごとの水分量を測定して、被験物質又は物品の接触後の水分分布を取得し、被験物質又は物品の接触後の水分分布と基準水分分布とを比較することで、被験物質又は物品が、ヒトの皮膚のバリア機能に与える影響を評価する、被験物質又は物品の評価方法であって、
水又は水溶液のヒトの皮膚への負荷前に、ヒトの皮膚の深さ毎の水分量を測定して初期水分分布を取得し、水又は水溶液のヒトの皮膚への負荷終了直後に、ヒトの皮膚の深さ毎の水分量の測定による水又は水溶液の負荷後の水分分布の取得と、指標物質を皮膚に適用して該指標物質の浸透度の測定を行い、初期水分分布から水又は水溶液の負荷終了直後の水分分布への変化と、指標物質の浸透度から、指標物質の浸透度と水又は水溶液の負荷量との関連性を決定し、前記関連性が変化したときのヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、前記基準水分分布とする、被験物質又は物品の評価方法に関する。 The present invention also provides a test substance by measuring the amount of water for each depth of human skin after contacting the test substance or article with human skin, and obtaining the moisture distribution after contact with the test substance or article. Or a test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin by comparing the moisture distribution after contact of the article with a reference moisture distribution,
Before applying water or aqueous solution to human skin, measure the amount of water at each depth of human skin to obtain the initial moisture distribution, and immediately after loading water or aqueous solution to human skin, Obtain water distribution after loading water or aqueous solution by measuring the amount of water at each skin depth, apply the indicator substance to the skin, measure the permeability of the indicator substance, and determine the water or aqueous solution from the initial moisture distribution. From the change in the moisture distribution immediately after the end of loading and the penetration of the indicator substance, the relationship between the penetration of the indicator substance and the load of water or aqueous solution is determined, and the human skin when the relation changes The present invention relates to a test substance or article evaluation method in which the moisture distribution after loading with water or an aqueous solution at each depth is used as the reference moisture distribution.
水又は水溶液のヒトの皮膚への負荷前に、ヒトの皮膚の深さ毎の水分量を測定して初期水分分布を取得し、水又は水溶液のヒトの皮膚への負荷終了直後に、ヒトの皮膚の深さ毎の水分量の測定による水又は水溶液の負荷後の水分分布の取得と、指標物質を皮膚に適用して該指標物質の浸透度の測定を行い、初期水分分布から水又は水溶液の負荷終了直後の水分分布への変化と、指標物質の浸透度から、指標物質の浸透度と水又は水溶液の負荷量との関連性を決定し、前記関連性が変化したときのヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、前記基準水分分布とする、被験物質又は物品の評価方法に関する。 The present invention also provides a test substance by measuring the amount of water for each depth of human skin after contacting the test substance or article with human skin, and obtaining the moisture distribution after contact with the test substance or article. Or a test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin by comparing the moisture distribution after contact of the article with a reference moisture distribution,
Before applying water or aqueous solution to human skin, measure the amount of water at each depth of human skin to obtain the initial moisture distribution, and immediately after loading water or aqueous solution to human skin, Obtain water distribution after loading water or aqueous solution by measuring the amount of water at each skin depth, apply the indicator substance to the skin, measure the permeability of the indicator substance, and determine the water or aqueous solution from the initial moisture distribution. From the change in the moisture distribution immediately after the end of loading and the penetration of the indicator substance, the relationship between the penetration of the indicator substance and the load of water or aqueous solution is determined, and the human skin when the relation changes The present invention relates to a test substance or article evaluation method in which the moisture distribution after loading with water or an aqueous solution at each depth is used as the reference moisture distribution.
さらに本発明は、被験物質又は物品が、ヒトの皮膚のバリア機能に与える影響を評価する方法であって、
被験物質又は物品のヒトの皮膚への接触前と接触後の皮膚の深さ毎の水分量を測定して水分分布を取得し、接触前と接触後の水分分布における、接触前の初期の角層厚よりも深部の水分量の差を指標として、被験物質又は物品がヒトの皮膚のバリア機能に与える影響を評価する、被験物質又は物品の評価方法に関する。 Furthermore, the present invention is a method for evaluating the effect of a test substance or article on the barrier function of human skin,
Obtain the moisture distribution by measuring the amount of moisture at each skin depth before and after contact of the test substance or article with human skin, and the initial angle before contact in the moisture distribution before and after contact. The present invention relates to a test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin using as an index the difference in moisture content deeper than the layer thickness.
被験物質又は物品のヒトの皮膚への接触前と接触後の皮膚の深さ毎の水分量を測定して水分分布を取得し、接触前と接触後の水分分布における、接触前の初期の角層厚よりも深部の水分量の差を指標として、被験物質又は物品がヒトの皮膚のバリア機能に与える影響を評価する、被験物質又は物品の評価方法に関する。 Furthermore, the present invention is a method for evaluating the effect of a test substance or article on the barrier function of human skin,
Obtain the moisture distribution by measuring the amount of moisture at each skin depth before and after contact of the test substance or article with human skin, and the initial angle before contact in the moisture distribution before and after contact. The present invention relates to a test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin using as an index the difference in moisture content deeper than the layer thickness.
本発明の上記及び他の特徴及び利点は、適宜添付の図面を参照して、下記の記載からより明らかになるであろう。
The above and other features and advantages of the present invention will become more apparent from the following description with reference to the accompanying drawings as appropriate.
本発明は、非侵襲的で、被験者に対して負担が少なく安全、簡便かつ短時間に、被験物質又は物品が皮膚バリア機能に与える影響を評価する、被験物質又は物品の評価方法を提供する。
The present invention provides a test substance or article evaluation method that evaluates the influence of a test substance or article on the skin barrier function in a non-invasive manner, with less burden on the subject, safe, simple, and in a short time.
従来より、皮膚の最表層に位置する角層の水分による膨潤状態(水和状態)が皮膚のバリア機能に影響し、それに伴い皮膚への物質の透過性も変化することが考えられていた。よって、被験物質又は物品が皮膚と接触することによって生じる角層の水和状態を把握できれば、被験物質又は物品が、皮膚のバリア機能にどのような影響を与えるかを、被験者に負担が少なく、様々な要因による影響を受けずに評価できるものと期待される。
しかし、皮膚への物質透過性変化と、その際の皮膚の水分量の増減や皮膚の深さ方向における水分分布との関係性について、詳細には解明されていなかった。 Conventionally, it has been considered that the swelling state (hydration state) of the stratum corneum located on the outermost layer of the skin affects the barrier function of the skin, and the permeability of the substance to the skin changes accordingly. Therefore, if it is possible to grasp the hydration state of the stratum corneum caused by the test substance or article coming into contact with the skin, the test substance or article has less burden on the subject as to how the test substance or article affects the barrier function of the skin, It is expected to be evaluated without being affected by various factors.
However, it has not been elucidated in detail about the relationship between the change in substance permeability to the skin and the increase / decrease of the moisture content of the skin and the moisture distribution in the depth direction of the skin.
しかし、皮膚への物質透過性変化と、その際の皮膚の水分量の増減や皮膚の深さ方向における水分分布との関係性について、詳細には解明されていなかった。 Conventionally, it has been considered that the swelling state (hydration state) of the stratum corneum located on the outermost layer of the skin affects the barrier function of the skin, and the permeability of the substance to the skin changes accordingly. Therefore, if it is possible to grasp the hydration state of the stratum corneum caused by the test substance or article coming into contact with the skin, the test substance or article has less burden on the subject as to how the test substance or article affects the barrier function of the skin, It is expected to be evaluated without being affected by various factors.
However, it has not been elucidated in detail about the relationship between the change in substance permeability to the skin and the increase / decrease of the moisture content of the skin and the moisture distribution in the depth direction of the skin.
そこで本発明者らは、生きたヒト皮膚における物質の透過性と、皮膚の最表面に位置する角層の水分量や深さ方向における水分分布との間の関連性について検討を行った。その結果、角層の水分量や深さ方向における水分分布と、皮膚への物質の透過性との間で高い関連性があることを生きたヒトにおいて初めて見出した。そして、被験物質又は物品をヒトの皮膚に接触させた前後での、非侵襲的に測定した生きたヒトにおける皮膚水分量や深さ方向における水分分布の変化から、被験物質又は物品が皮膚のバリア機能に与える影響を評価できることを見い出した。
本発明はこれらの知見に基づき完成されるに至ったものである。 Therefore, the present inventors examined the relationship between the permeability of a substance in living human skin and the moisture content of the stratum corneum located on the outermost surface of the skin and the moisture distribution in the depth direction. As a result, we found for the first time in living humans that there is a high correlation between the moisture content of the stratum corneum and the moisture distribution in the depth direction and the permeability of substances to the skin. Then, the test substance or article becomes a skin barrier from the change in the skin moisture content or the moisture distribution in the depth direction in a living human measured non-invasively before and after the test substance or article is brought into contact with the human skin. It was found that the effect on the function can be evaluated.
The present invention has been completed based on these findings.
本発明はこれらの知見に基づき完成されるに至ったものである。 Therefore, the present inventors examined the relationship between the permeability of a substance in living human skin and the moisture content of the stratum corneum located on the outermost surface of the skin and the moisture distribution in the depth direction. As a result, we found for the first time in living humans that there is a high correlation between the moisture content of the stratum corneum and the moisture distribution in the depth direction and the permeability of substances to the skin. Then, the test substance or article becomes a skin barrier from the change in the skin moisture content or the moisture distribution in the depth direction in a living human measured non-invasively before and after the test substance or article is brought into contact with the human skin. It was found that the effect on the function can be evaluated.
The present invention has been completed based on these findings.
本発明では、皮膚の水分量や水分分布と皮膚のバリア機能との関連性に基づき、被験物質又は物品の接触前後の皮膚の水分量や水分分布から、皮膚のバリア機能を評価する。よって本発明によれば、被験物質又は物品が皮膚に与える影響を安全、簡便かつ短時間に知ることができる。
In the present invention, the skin barrier function is evaluated from the moisture content and moisture distribution of the skin before and after contact with the test substance or article based on the relationship between the moisture content and moisture distribution of the skin and the barrier function of the skin. Therefore, according to the present invention, the influence of the test substance or article on the skin can be known safely, simply and in a short time.
本発明では、図1に示すように、被験物質又は物品をヒトの皮膚に接触させ(S1)、被験物質又は物品の接触前後に測定した皮膚の水分分量又は水分分布の変化量や、接触後の皮膚の水分分布を、基準値又は基準水分分布と比較し(S2)、基準値又は基準水分分布との比較結果から、被験物質又は物品が皮膚のバリア機能に与える影響を評価する(S3)。ここで、皮膚のバリア機能に与える影響の解析と評価は、皮膚の水分量又は水分分布の変化量と、皮膚のバリア機能との関連性に基づいて行われる。
以下、本発明について詳細に説明する。 In the present invention, as shown in FIG. 1, the test substance or article is brought into contact with human skin (S1), the amount of change in the moisture content or moisture distribution of the skin measured before and after contact with the test substance or article, Compare the moisture distribution of the skin with the reference value or reference moisture distribution (S2), and evaluate the effect of the test substance or article on the skin barrier function from the comparison result with the reference value or reference moisture distribution (S3) . Here, analysis and evaluation of the influence on the skin barrier function are performed based on the relationship between the amount of moisture in the skin or the distribution of moisture distribution and the barrier function of the skin.
Hereinafter, the present invention will be described in detail.
以下、本発明について詳細に説明する。 In the present invention, as shown in FIG. 1, the test substance or article is brought into contact with human skin (S1), the amount of change in the moisture content or moisture distribution of the skin measured before and after contact with the test substance or article, Compare the moisture distribution of the skin with the reference value or reference moisture distribution (S2), and evaluate the effect of the test substance or article on the skin barrier function from the comparison result with the reference value or reference moisture distribution (S3) . Here, analysis and evaluation of the influence on the skin barrier function are performed based on the relationship between the amount of moisture in the skin or the distribution of moisture distribution and the barrier function of the skin.
Hereinafter, the present invention will be described in detail.
本明細書において「バリア機能」とは、皮膚外部から内部への物質の浸透性を指す。
本明細書において「被験物質」とは、皮膚との接触や皮膚への適用により、皮膚内へ浸透する物質(例えば、化合物、化学物質、色素、生体由来の分泌物や***物、保湿剤、外用剤、化粧料)をいう。なお「被験物質」及び後述する「指標物質」には、水溶性成分や疎水性成分も包含される。物質の具体例としては、水若しくは水蒸気、生理食塩水、リン酸緩衝生理食塩水、ニコチン酸メチル、ニコチン酸ヘキシル、乳酸、黄色4号(タートラジン)、赤色215号(ローダミンBステアレート)、カフェイン、ユビナール、カロテノイド、レチノール、尿、汗、経血、便、擬似尿、擬似経血、擬似便、並びに、皮膚保湿剤、皮膚外用剤、皮膚化粧料若しくは蒸気温熱具に含まれる有効成分や薬効成分が挙げられる。 As used herein, “barrier function” refers to the permeability of a substance from the outside to the inside of the skin.
In the present specification, the “test substance” means a substance (for example, a compound, a chemical substance, a pigment, a secretion or excrement derived from a living body, a moisturizing agent, or the like that penetrates into the skin by contact with the skin or application to the skin. External preparations, cosmetics). The “test substance” and the “indicator substance” described later include water-soluble components and hydrophobic components. Specific examples of substances include water or water vapor, physiological saline, phosphate buffered saline, methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4 (tartrazine), red No. 215 (rhodamine B stearate), cafe In, ubinal, carotenoids, retinol, urine, sweat, menstrual blood, feces, simulated urine, simulated menstrual blood, simulated feces, and active ingredients contained in skin moisturizers, topical skin preparations, skin cosmetics or steam heating devices Medicinal ingredients are listed.
本明細書において「被験物質」とは、皮膚との接触や皮膚への適用により、皮膚内へ浸透する物質(例えば、化合物、化学物質、色素、生体由来の分泌物や***物、保湿剤、外用剤、化粧料)をいう。なお「被験物質」及び後述する「指標物質」には、水溶性成分や疎水性成分も包含される。物質の具体例としては、水若しくは水蒸気、生理食塩水、リン酸緩衝生理食塩水、ニコチン酸メチル、ニコチン酸ヘキシル、乳酸、黄色4号(タートラジン)、赤色215号(ローダミンBステアレート)、カフェイン、ユビナール、カロテノイド、レチノール、尿、汗、経血、便、擬似尿、擬似経血、擬似便、並びに、皮膚保湿剤、皮膚外用剤、皮膚化粧料若しくは蒸気温熱具に含まれる有効成分や薬効成分が挙げられる。 As used herein, “barrier function” refers to the permeability of a substance from the outside to the inside of the skin.
In the present specification, the “test substance” means a substance (for example, a compound, a chemical substance, a pigment, a secretion or excrement derived from a living body, a moisturizing agent, or the like that penetrates into the skin by contact with the skin or application to the skin. External preparations, cosmetics). The “test substance” and the “indicator substance” described later include water-soluble components and hydrophobic components. Specific examples of substances include water or water vapor, physiological saline, phosphate buffered saline, methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4 (tartrazine), red No. 215 (rhodamine B stearate), cafe In, ubinal, carotenoids, retinol, urine, sweat, menstrual blood, feces, simulated urine, simulated menstrual blood, simulated feces, and active ingredients contained in skin moisturizers, topical skin preparations, skin cosmetics or steam heating devices Medicinal ingredients are listed.
まず、被験物質又は物品の皮膚への接触前後で、バリア機能の評価を行う部位の皮膚水分量又は水分分布を測定する。皮膚水分量又は水分分布の測定は、非侵襲的に測定することが好ましい。この場合、被験者に対して負担が少なく、安全に皮膚のバリア機能を評価できる。
角層内の水分量又は水分分布を非侵襲的に測定する方法は、常法から適宜選択することができる。例えば、ラマン分光法、皮膚静電容量測定手段、高周波電流測定手段、赤外分光法、又は近赤外分光法などを用いる方法が挙げられる。 First, before and after contact of the test substance or article with the skin, the skin moisture content or moisture distribution at the site where the barrier function is evaluated is measured. The measurement of the skin moisture content or moisture distribution is preferably performed noninvasively. In this case, the burden on the subject is small, and the skin barrier function can be safely evaluated.
The method for noninvasively measuring the moisture content or moisture distribution in the stratum corneum can be appropriately selected from conventional methods. Examples thereof include a method using Raman spectroscopy, skin capacitance measurement means, high-frequency current measurement means, infrared spectroscopy, or near infrared spectroscopy.
角層内の水分量又は水分分布を非侵襲的に測定する方法は、常法から適宜選択することができる。例えば、ラマン分光法、皮膚静電容量測定手段、高周波電流測定手段、赤外分光法、又は近赤外分光法などを用いる方法が挙げられる。 First, before and after contact of the test substance or article with the skin, the skin moisture content or moisture distribution at the site where the barrier function is evaluated is measured. The measurement of the skin moisture content or moisture distribution is preferably performed noninvasively. In this case, the burden on the subject is small, and the skin barrier function can be safely evaluated.
The method for noninvasively measuring the moisture content or moisture distribution in the stratum corneum can be appropriately selected from conventional methods. Examples thereof include a method using Raman spectroscopy, skin capacitance measurement means, high-frequency current measurement means, infrared spectroscopy, or near infrared spectroscopy.
本発明では、皮膚に対して被験物質又は物品を負荷した場合の皮膚水分量又は水分分布の変化を、ラマン分光法、皮膚静電容量測定手段などに従い皮膚水分量又は水分分布を測定できる装置を用いて測定する。そして、その変化量から、該物質又は物品が皮膚のバリア機能に及ぼす影響を評価する。
後述の実施例で示したように、ヒトの皮膚に過剰な水分を負荷すると物質透過性が亢進し、皮膚のバリア機能は低下する。
このような知見に対して、本発明者らは、皮膚に対して被験物質又は物品を負荷した場合の皮膚水分量又は水分分布の変化値や基準水分分布を基準に、皮膚のバリア機能を評価できることをはじめて見出した。後述の実施例でも示すように、皮膚の水分量がある一定値までは物質浸透性は亢進しない。しかし、皮膚の水分量が一定値以上増加すると、物質浸透性が亢進する。なお、後述の実施例では、指標物質の代表例として、水溶性物質であるニコチン酸メチルの浸透性の評価を行った。水溶性物質は皮膚の水分量が増えると皮膚内へ物質が分配されやすくなり、皮膚により浸透する可能性が考えられる。 In the present invention, there is provided an apparatus capable of measuring skin moisture content or moisture distribution in accordance with Raman spectroscopy, skin capacitance measuring means, etc., when the test substance or article is loaded on the skin. Use to measure. Then, the influence of the substance or article on the skin barrier function is evaluated from the amount of change.
As shown in Examples described later, when excessive moisture is applied to human skin, the substance permeability is enhanced and the skin barrier function is lowered.
In response to such findings, the present inventors evaluated the skin barrier function based on the skin moisture content or the change in the moisture distribution when the test substance or article is loaded on the skin or the reference moisture distribution. I found what I could do for the first time. As shown in the examples described later, the substance permeability does not increase until the skin moisture content reaches a certain value. However, when the moisture content of the skin increases above a certain value, the substance permeability increases. In the examples described later, the permeability of methyl nicotinate, which is a water-soluble substance, was evaluated as a representative example of the indicator substance. A water-soluble substance is likely to be distributed into the skin as the amount of moisture in the skin increases, and may penetrate into the skin.
後述の実施例で示したように、ヒトの皮膚に過剰な水分を負荷すると物質透過性が亢進し、皮膚のバリア機能は低下する。
このような知見に対して、本発明者らは、皮膚に対して被験物質又は物品を負荷した場合の皮膚水分量又は水分分布の変化値や基準水分分布を基準に、皮膚のバリア機能を評価できることをはじめて見出した。後述の実施例でも示すように、皮膚の水分量がある一定値までは物質浸透性は亢進しない。しかし、皮膚の水分量が一定値以上増加すると、物質浸透性が亢進する。なお、後述の実施例では、指標物質の代表例として、水溶性物質であるニコチン酸メチルの浸透性の評価を行った。水溶性物質は皮膚の水分量が増えると皮膚内へ物質が分配されやすくなり、皮膚により浸透する可能性が考えられる。 In the present invention, there is provided an apparatus capable of measuring skin moisture content or moisture distribution in accordance with Raman spectroscopy, skin capacitance measuring means, etc., when the test substance or article is loaded on the skin. Use to measure. Then, the influence of the substance or article on the skin barrier function is evaluated from the amount of change.
As shown in Examples described later, when excessive moisture is applied to human skin, the substance permeability is enhanced and the skin barrier function is lowered.
In response to such findings, the present inventors evaluated the skin barrier function based on the skin moisture content or the change in the moisture distribution when the test substance or article is loaded on the skin or the reference moisture distribution. I found what I could do for the first time. As shown in the examples described later, the substance permeability does not increase until the skin moisture content reaches a certain value. However, when the moisture content of the skin increases above a certain value, the substance permeability increases. In the examples described later, the permeability of methyl nicotinate, which is a water-soluble substance, was evaluated as a representative example of the indicator substance. A water-soluble substance is likely to be distributed into the skin as the amount of moisture in the skin increases, and may penetrate into the skin.
本発明において、被験物質又は物品が、皮膚バリア機能に与える影響を評価するための基準値や基準水分分布は適宜設定することができる。以下、本発明における基準値や基準水分分布の設定方法ついて、具体例に基づき説明する。しかし、本発明はこれに制限するものではない。
まず、水の負荷量を変えて皮膚への水負荷試験を行い、各水負荷量での皮膚の水分量又は水分分布を常法により測定する。次に、測定した皮膚の水分量又は水分分布と、皮膚のバリア機能との関連付けを行う。 In the present invention, the reference value and the reference moisture distribution for evaluating the influence of the test substance or article on the skin barrier function can be set as appropriate. Hereinafter, the setting method of the reference value and the reference moisture distribution in the present invention will be described based on specific examples. However, the present invention is not limited to this.
First, the water load test is performed on the skin by changing the water load, and the moisture content or moisture distribution of the skin at each water load is measured by a conventional method. Next, the measured moisture content or moisture distribution of the skin is associated with the barrier function of the skin.
まず、水の負荷量を変えて皮膚への水負荷試験を行い、各水負荷量での皮膚の水分量又は水分分布を常法により測定する。次に、測定した皮膚の水分量又は水分分布と、皮膚のバリア機能との関連付けを行う。 In the present invention, the reference value and the reference moisture distribution for evaluating the influence of the test substance or article on the skin barrier function can be set as appropriate. Hereinafter, the setting method of the reference value and the reference moisture distribution in the present invention will be described based on specific examples. However, the present invention is not limited to this.
First, the water load test is performed on the skin by changing the water load, and the moisture content or moisture distribution of the skin at each water load is measured by a conventional method. Next, the measured moisture content or moisture distribution of the skin is associated with the barrier function of the skin.
本発明の第1の実施態様において、皮膚の水分量又は水分分布と皮膚のバリア機能との関連付けは、皮膚のバリア機能に変化が確認されるときの皮膚の水分分量又は水分分布を「基準値」として設定することで行う。
「基準値」の設定方法としては、皮膚外部から内部への前記指標物質の浸透性から設定する方法が挙げられる。前述の皮膚への水負荷試験に続いて皮膚への指標物質の負荷試験を行って指標物質の浸透性を常法により測定し、指標物質の浸透性に変化がみられたときの皮膚の水分量又は水分分布を「基準値」とすることができる。なお、本実施態様においては、負荷した水分量と、指標物質の浸透性との間で関連に変化が確認されたときや、指標物質の浸透性が一定の状態に達したときを基準値とすることができる。 In the first embodiment of the present invention, the association between the moisture content or moisture distribution of the skin and the barrier function of the skin is defined as “the reference value of the moisture content or moisture distribution of the skin when a change is confirmed in the skin barrier function. ”Is set.
Examples of the method for setting the “reference value” include a method for setting from the permeability of the indicator substance from the outside to the inside of the skin. Following the water load test on the skin described above, the load test of the index substance to the skin was conducted to measure the permeability of the index substance by a conventional method, and the moisture of the skin when the change in the permeability of the index substance was observed The amount or moisture distribution can be a “reference value”. In this embodiment, when a change is confirmed in relation between the amount of moisture loaded and the permeability of the indicator substance, or when the permeability of the indicator substance reaches a certain state, can do.
「基準値」の設定方法としては、皮膚外部から内部への前記指標物質の浸透性から設定する方法が挙げられる。前述の皮膚への水負荷試験に続いて皮膚への指標物質の負荷試験を行って指標物質の浸透性を常法により測定し、指標物質の浸透性に変化がみられたときの皮膚の水分量又は水分分布を「基準値」とすることができる。なお、本実施態様においては、負荷した水分量と、指標物質の浸透性との間で関連に変化が確認されたときや、指標物質の浸透性が一定の状態に達したときを基準値とすることができる。 In the first embodiment of the present invention, the association between the moisture content or moisture distribution of the skin and the barrier function of the skin is defined as “the reference value of the moisture content or moisture distribution of the skin when a change is confirmed in the skin barrier function. ”Is set.
Examples of the method for setting the “reference value” include a method for setting from the permeability of the indicator substance from the outside to the inside of the skin. Following the water load test on the skin described above, the load test of the index substance to the skin was conducted to measure the permeability of the index substance by a conventional method, and the moisture of the skin when the change in the permeability of the index substance was observed The amount or moisture distribution can be a “reference value”. In this embodiment, when a change is confirmed in relation between the amount of moisture loaded and the permeability of the indicator substance, or when the permeability of the indicator substance reaches a certain state, can do.
本発明の第2の実施態様において、皮膚の水分量又は水分分布と皮膚のバリア機能との関連付けは、指標物質の浸透度と水又は水溶液の負荷量との関連性が変化したときの、ヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、「基準水分分布」として設定することで行う。
本実施態様ではまず、水又は水溶液のヒトの皮膚への負荷前に、ヒトの皮膚の深さ毎の水分量を測定して初期水分分布を取得する。
さらに、水又は水溶液をヒトの皮膚に一定時間負荷し、負荷終了直後にヒトの皮膚の深さ毎の水分量を測定して水又は水溶液の負荷後の水分分布を取得する。続けて指標物質を皮膚に適用して該指標物質の浸透度を測定する。
そして、指標物質の浸透度と水又は水溶液の負荷量との関連性が変化するまで、水又は水溶液の負荷量を増加又は減少させながら前記測定を繰り返す。この測定結果に基づいて、指標物質の浸透度と水又は水溶液の負荷量との関連性が変化したときのヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、前記基準水分分布とすることができる。 In the second embodiment of the present invention, the association between the moisture content or moisture distribution of the skin and the barrier function of the skin is the human when the association between the permeability of the indicator substance and the load of water or aqueous solution is changed. The water distribution after loading with water or an aqueous solution at each skin depth is set as the “reference water distribution”.
In this embodiment, first, before loading water or an aqueous solution on human skin, the amount of water at each depth of the human skin is measured to obtain an initial moisture distribution.
Furthermore, water or an aqueous solution is loaded onto the human skin for a certain period of time, and immediately after the end of the loading, the moisture content at each depth of the human skin is measured to obtain the moisture distribution after loading with the water or aqueous solution. Subsequently, the indicator substance is applied to the skin and the penetration degree of the indicator substance is measured.
Then, the measurement is repeated while increasing or decreasing the load of water or aqueous solution until the relationship between the permeability of the indicator substance and the load of water or aqueous solution changes. Based on the measurement results, the water distribution after loading of water or aqueous solution at each depth of human skin when the relationship between the permeability of the indicator substance and the load of water or aqueous solution changes is determined as the reference water content. It can be a distribution.
本実施態様ではまず、水又は水溶液のヒトの皮膚への負荷前に、ヒトの皮膚の深さ毎の水分量を測定して初期水分分布を取得する。
さらに、水又は水溶液をヒトの皮膚に一定時間負荷し、負荷終了直後にヒトの皮膚の深さ毎の水分量を測定して水又は水溶液の負荷後の水分分布を取得する。続けて指標物質を皮膚に適用して該指標物質の浸透度を測定する。
そして、指標物質の浸透度と水又は水溶液の負荷量との関連性が変化するまで、水又は水溶液の負荷量を増加又は減少させながら前記測定を繰り返す。この測定結果に基づいて、指標物質の浸透度と水又は水溶液の負荷量との関連性が変化したときのヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、前記基準水分分布とすることができる。 In the second embodiment of the present invention, the association between the moisture content or moisture distribution of the skin and the barrier function of the skin is the human when the association between the permeability of the indicator substance and the load of water or aqueous solution is changed. The water distribution after loading with water or an aqueous solution at each skin depth is set as the “reference water distribution”.
In this embodiment, first, before loading water or an aqueous solution on human skin, the amount of water at each depth of the human skin is measured to obtain an initial moisture distribution.
Furthermore, water or an aqueous solution is loaded onto the human skin for a certain period of time, and immediately after the end of the loading, the moisture content at each depth of the human skin is measured to obtain the moisture distribution after loading with the water or aqueous solution. Subsequently, the indicator substance is applied to the skin and the penetration degree of the indicator substance is measured.
Then, the measurement is repeated while increasing or decreasing the load of water or aqueous solution until the relationship between the permeability of the indicator substance and the load of water or aqueous solution changes. Based on the measurement results, the water distribution after loading of water or aqueous solution at each depth of human skin when the relationship between the permeability of the indicator substance and the load of water or aqueous solution changes is determined as the reference water content. It can be a distribution.
本発明の第1及び第2の実施態様において、皮膚への浸透性を評価するための指標物質は、適宜選択することができる。例えば、ニコチン酸メチル、ニコチン酸ヘキシル、乳酸、黄色4号、赤色215号、カフェイン、ユビナール、カロテノイド、レチノール、並びに、皮膚保湿剤、皮膚外用剤、皮膚化粧料若しくは蒸気温熱具に含まれる有効成分や薬効成分が挙げられる。
In the first and second embodiments of the present invention, the indicator substance for evaluating the permeability to the skin can be appropriately selected. For example, methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4, red No. 215, caffeine, ubinal, carotenoid, retinol, and effective in skin moisturizers, skin external preparations, skin cosmetics or steam heating devices Ingredients and medicinal ingredients.
本発明の第3の実施態様では、初期の角層厚よりも深部の水分量と、皮膚のバリア機能とを関連付ける。
本実施態様では、被験物質又は物品をヒトの皮膚に接触させる前と接触させた後の皮膚の深さ毎の水分量を測定して水分分布を取得する。そして、接触前後の水分分布を比較し、接触前の初期の角層厚よりも深部の水分量の差を指標として、被験物質又は物品の評価を行う。 In the third embodiment of the present invention, the moisture content deeper than the initial stratum corneum thickness is associated with the skin barrier function.
In this embodiment, the moisture distribution is obtained by measuring the amount of moisture at each skin depth before and after the test substance or article is brought into contact with human skin. Then, the moisture distribution before and after contact is compared, and the test substance or article is evaluated using as an index the difference in moisture content deeper than the initial stratum corneum thickness before contact.
本実施態様では、被験物質又は物品をヒトの皮膚に接触させる前と接触させた後の皮膚の深さ毎の水分量を測定して水分分布を取得する。そして、接触前後の水分分布を比較し、接触前の初期の角層厚よりも深部の水分量の差を指標として、被験物質又は物品の評価を行う。 In the third embodiment of the present invention, the moisture content deeper than the initial stratum corneum thickness is associated with the skin barrier function.
In this embodiment, the moisture distribution is obtained by measuring the amount of moisture at each skin depth before and after the test substance or article is brought into contact with human skin. Then, the moisture distribution before and after contact is compared, and the test substance or article is evaluated using as an index the difference in moisture content deeper than the initial stratum corneum thickness before contact.
本発明では、角層表層から顆粒層に達するまでのいずれかの領域又は部位での水分量又は水分分布を測定してもよい。あるいは、本発明で用いる水分量又は水分分布の測定装置の特性、例えば皮膚の内部に照射した励起波長によるラマン散乱の強度などを考慮し、皮膚表面から所定の深さまでの領域の水分量若しくは水分分布、又は皮膚表面から所定の深さの部位の水分量を測定してもよい。例えば、皮膚のバリア機能を評価するために、皮膚表面から初期の角層厚における水分量又は水分分布を測定することが好ましい。
In the present invention, the amount of moisture or the moisture distribution in any region or part from the stratum corneum surface layer to the granular layer may be measured. Or, considering the characteristics of the moisture content or moisture distribution measuring device used in the present invention, for example, the intensity of Raman scattering due to the excitation wavelength applied to the inside of the skin, the moisture content or moisture in the region from the skin surface to a predetermined depth You may measure distribution or the moisture content of the site | part of a predetermined depth from the skin surface. For example, in order to evaluate the barrier function of the skin, it is preferable to measure the moisture content or moisture distribution at the initial stratum corneum thickness from the skin surface.
なお本明細書において、「初期の角層厚」とは、水分、被験物質又は物品負荷前の角層の厚さを指す。なお、ラマンスペクトルからの初期の角層厚の測定は、IFSCC Magazine, 2009, vol. 12(1), p. 9-15に記載の方法に準じて行うことができる。常法に従い測定したラマンスペクトルから各深さにおける水分量を算出し、深さ方向に対して一定距離(例えば1μm)ごとに近似的に算出した水分量の数値をプロットしたグラフを作成し、水分分布プロファイルを取得する。そして、取得した水分プロファイルに対して2本の直線で近似し、その交点までの深さを、角層表層からの角層厚と定義することができる。
また、ラマンスペクトルで皮膚の水分量を測定する場合、タンパク質のCH伸縮振動由来の信号強度は、対物レンズの窓材と皮膚の境界よりやや皮膚側で最大値を示した後に、深部にいくほど信号強度が低下する。皮膚では深部にいくほど光の拡散などによる信号の減衰が生じるためである。窓材にはタンパク質のCH伸縮振動由来の信号は存在しないため、レーザーの焦点が窓材の内にあるときはCH伸縮振動由来の信号強度は0になる。一方、皮膚内にレーザーの焦点があるときのCH伸縮振動由来の信号強度を100とすると、窓材と皮膚の界面に焦点がきた場合、CH伸縮振動由来の信号強度は50となる。そこで、CH伸縮振動由来の信号強度が最大値の半値を示すところを皮膚と窓材の境界と定義している。ただし、実際には窓材にはCH伸縮振動由来の信号強度が弱いながらも検出されることがある。そのため、特開2012-50739号公報に記載の方法に従って水分量を計算、解析すると、高い水分量を示すことになり、皮膚と窓材の界面、すなわち皮膚の最表面の水分量に関しては適切ではないデータであるということが理論上明らかとなっている。さらに、本条件に用いた共焦点ラマン分光装置の深さ方向の空間分解能は2μmで、解析の際に1μm毎の水分量を補間して算出している。よって、皮膚表面の水分量は、データ解析からは省くことが好ましい。また、共焦点ラマンの装置によってそれぞれ分解能が異なることを考慮し、皮膚表面水分量を求める際には、本発明においては、実際の皮膚表面から最低5μmまでのデータを省き、皮膚表面から5μm以上の部位をデータ解析上の「皮膚表面」とすることが好ましく、実際の皮膚表面から最低2μmまでのデータを省き、皮膚表面から2μm以上の部位をデータ解析上の「皮膚表面」とすることがより好ましく、実際の皮膚表面から2μmまでのデータを省き、皮膚表面から2μmの部位をデータ解析上の「皮膚表面」とすることがさらに好ましい。
共焦点ラマン分光顕微鏡を用いた測定で得られたラマンスペクトルから水分量を近似的に算出する方法は、特開2012-50739号公報に記載の方法を参照して行うことができる。 In the present specification, the “initial stratum corneum thickness” refers to the thickness of the stratum corneum before moisture, a test substance or an article is loaded. The initial stratum corneum thickness from the Raman spectrum can be measured according to the method described in IFSCC Magazine, 2009, vol. 12 (1), p. 9-15. Calculate the moisture content at each depth from the Raman spectrum measured according to the usual method, and create a graph plotting the numerical value of the moisture content approximately calculated for each fixed distance (eg 1 μm) in the depth direction. Get distribution profile. Then, the obtained moisture profile is approximated by two straight lines, and the depth to the intersection can be defined as the stratum corneum thickness from the stratum corneum surface layer.
Moreover, when measuring the moisture content of the skin with a Raman spectrum, the signal intensity derived from the CH stretching vibration of the protein shows a maximum value on the skin side slightly from the boundary between the window material of the objective lens and the skin, and then goes deeper. Signal strength decreases. This is because, in the skin, the signal is attenuated by the diffusion of light as it goes deeper. Since there is no signal derived from protein CH stretching vibration in the window material, the signal intensity derived from CH stretching vibration is 0 when the focal point of the laser is within the window material. On the other hand, when the signal intensity derived from the CH stretching vibration when the laser is focused in the skin is 100, the signal intensity derived from the CH stretching vibration is 50 when the focus comes to the interface between the window material and the skin. Therefore, the point where the signal intensity derived from the CH stretching vibration shows the maximum half value is defined as the boundary between the skin and the window material. In practice, however, the window material may be detected even though the signal intensity derived from the CH stretching vibration is weak. Therefore, when the moisture content is calculated and analyzed according to the method described in JP2012-50739A, the moisture content will be high, and the interface between the skin and the window material, that is, the moisture content on the outermost surface of the skin is not appropriate. It is theoretically clear that there is no data. Further, the spatial resolution in the depth direction of the confocal Raman spectroscopic device used in this condition is 2 μm, and the water amount for each 1 μm is interpolated and calculated in the analysis. Therefore, it is preferable to omit the moisture content on the skin surface from the data analysis. In consideration of the fact that the resolution varies depending on the confocal Raman device, in the present invention, when determining the moisture content on the skin surface, the data from the actual skin surface to at least 5 μm is omitted, and the skin surface is not less than 5 μm. It is preferable that the part of the skin is the “skin surface” in the data analysis, the data of at least 2 μm from the actual skin surface is omitted, and the part of 2 μm or more from the skin surface is the “skin surface” in the data analysis More preferably, it is more preferable to omit the data from the actual skin surface to 2 μm, and to set the portion of 2 μm from the skin surface as the “skin surface” in the data analysis.
A method for approximately calculating the amount of water from a Raman spectrum obtained by measurement using a confocal Raman spectroscopic microscope can be performed with reference to the method described in JP 2012-50739 A.
また、ラマンスペクトルで皮膚の水分量を測定する場合、タンパク質のCH伸縮振動由来の信号強度は、対物レンズの窓材と皮膚の境界よりやや皮膚側で最大値を示した後に、深部にいくほど信号強度が低下する。皮膚では深部にいくほど光の拡散などによる信号の減衰が生じるためである。窓材にはタンパク質のCH伸縮振動由来の信号は存在しないため、レーザーの焦点が窓材の内にあるときはCH伸縮振動由来の信号強度は0になる。一方、皮膚内にレーザーの焦点があるときのCH伸縮振動由来の信号強度を100とすると、窓材と皮膚の界面に焦点がきた場合、CH伸縮振動由来の信号強度は50となる。そこで、CH伸縮振動由来の信号強度が最大値の半値を示すところを皮膚と窓材の境界と定義している。ただし、実際には窓材にはCH伸縮振動由来の信号強度が弱いながらも検出されることがある。そのため、特開2012-50739号公報に記載の方法に従って水分量を計算、解析すると、高い水分量を示すことになり、皮膚と窓材の界面、すなわち皮膚の最表面の水分量に関しては適切ではないデータであるということが理論上明らかとなっている。さらに、本条件に用いた共焦点ラマン分光装置の深さ方向の空間分解能は2μmで、解析の際に1μm毎の水分量を補間して算出している。よって、皮膚表面の水分量は、データ解析からは省くことが好ましい。また、共焦点ラマンの装置によってそれぞれ分解能が異なることを考慮し、皮膚表面水分量を求める際には、本発明においては、実際の皮膚表面から最低5μmまでのデータを省き、皮膚表面から5μm以上の部位をデータ解析上の「皮膚表面」とすることが好ましく、実際の皮膚表面から最低2μmまでのデータを省き、皮膚表面から2μm以上の部位をデータ解析上の「皮膚表面」とすることがより好ましく、実際の皮膚表面から2μmまでのデータを省き、皮膚表面から2μmの部位をデータ解析上の「皮膚表面」とすることがさらに好ましい。
共焦点ラマン分光顕微鏡を用いた測定で得られたラマンスペクトルから水分量を近似的に算出する方法は、特開2012-50739号公報に記載の方法を参照して行うことができる。 In the present specification, the “initial stratum corneum thickness” refers to the thickness of the stratum corneum before moisture, a test substance or an article is loaded. The initial stratum corneum thickness from the Raman spectrum can be measured according to the method described in IFSCC Magazine, 2009, vol. 12 (1), p. 9-15. Calculate the moisture content at each depth from the Raman spectrum measured according to the usual method, and create a graph plotting the numerical value of the moisture content approximately calculated for each fixed distance (eg 1 μm) in the depth direction. Get distribution profile. Then, the obtained moisture profile is approximated by two straight lines, and the depth to the intersection can be defined as the stratum corneum thickness from the stratum corneum surface layer.
Moreover, when measuring the moisture content of the skin with a Raman spectrum, the signal intensity derived from the CH stretching vibration of the protein shows a maximum value on the skin side slightly from the boundary between the window material of the objective lens and the skin, and then goes deeper. Signal strength decreases. This is because, in the skin, the signal is attenuated by the diffusion of light as it goes deeper. Since there is no signal derived from protein CH stretching vibration in the window material, the signal intensity derived from CH stretching vibration is 0 when the focal point of the laser is within the window material. On the other hand, when the signal intensity derived from the CH stretching vibration when the laser is focused in the skin is 100, the signal intensity derived from the CH stretching vibration is 50 when the focus comes to the interface between the window material and the skin. Therefore, the point where the signal intensity derived from the CH stretching vibration shows the maximum half value is defined as the boundary between the skin and the window material. In practice, however, the window material may be detected even though the signal intensity derived from the CH stretching vibration is weak. Therefore, when the moisture content is calculated and analyzed according to the method described in JP2012-50739A, the moisture content will be high, and the interface between the skin and the window material, that is, the moisture content on the outermost surface of the skin is not appropriate. It is theoretically clear that there is no data. Further, the spatial resolution in the depth direction of the confocal Raman spectroscopic device used in this condition is 2 μm, and the water amount for each 1 μm is interpolated and calculated in the analysis. Therefore, it is preferable to omit the moisture content on the skin surface from the data analysis. In consideration of the fact that the resolution varies depending on the confocal Raman device, in the present invention, when determining the moisture content on the skin surface, the data from the actual skin surface to at least 5 μm is omitted, and the skin surface is not less than 5 μm. It is preferable that the part of the skin is the “skin surface” in the data analysis, the data of at least 2 μm from the actual skin surface is omitted, and the part of 2 μm or more from the skin surface is the “skin surface” in the data analysis More preferably, it is more preferable to omit the data from the actual skin surface to 2 μm, and to set the portion of 2 μm from the skin surface as the “skin surface” in the data analysis.
A method for approximately calculating the amount of water from a Raman spectrum obtained by measurement using a confocal Raman spectroscopic microscope can be performed with reference to the method described in JP 2012-50739 A.
ラマン分光法にて測定した初期の角層厚は通常、およそ10μm~20μm、平均で約13μmである。そこで、本発明において、皮膚の水分量は、実際の皮膚表面からの深さ2~40μmの深さまで、好ましくは皮膚表面からの深さ2~35μmの深さまで、より好ましくは皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、を測定し、解析するのが好ましい。
The initial stratum corneum thickness measured by Raman spectroscopy is usually about 10 μm to 20 μm and an average of about 13 μm. Therefore, in the present invention, the moisture content of the skin is from 2 to 40 μm deep from the actual skin surface, preferably from 2 to 35 μm deep from the skin surface, more preferably from the skin surface. It is preferable to measure and analyze up to a depth of 2 to 20 μm, more preferably a depth of 9 to 15 μm from the skin surface, more preferably a depth of about 13 μm from the skin surface.
後述の実施例でも示すように、ラマン分光法や皮膚静電容量測定手段などで測定した皮膚の水分量又は水分分布と、皮膚のバリア機能とは強い関連性を有する。具体的には、被験物質又は物品の接触により皮膚の水分量の変化量が一定値以上増加すると、ニコチン酸メチルなどの物質の皮膚への浸透性が亢進する。
本発明では、上述のような皮膚の水分量又は水分分布と、バリア機能との関連性に基づいて、非侵襲的に測定した皮膚の水分量又は水分分布の変化量から、被験物質又は物品が、皮膚のバリア機能に及ぼす影響を評価する。具体的には、被験物質又は物品の接触後の皮膚の水分量が、所定の基準値以上の変化量で増加した場合、バリア機能を低下させる(例えば、皮膚外部から内部への物質の浸透性を亢進させる)と評価することができる。一方、被験物質又は物品の接触後の皮膚の水分量が、基準値より小さい変化量で変化した場合、バリア機能は変化してないと評価することができる。 As shown in Examples described later, the skin moisture amount or moisture distribution measured by Raman spectroscopy, skin capacitance measuring means, or the like has a strong relationship with the skin barrier function. Specifically, when the amount of change in the amount of moisture in the skin increases by a certain value or more due to contact with the test substance or article, the permeability of substances such as methyl nicotinate to the skin is enhanced.
In the present invention, based on the relationship between the moisture content or moisture distribution of the skin as described above and the barrier function, the test substance or article is obtained from the amount of change in the moisture content or moisture distribution measured noninvasively. Evaluate the effects on skin barrier function. Specifically, when the amount of moisture in the skin after contact with the test substance or article increases with a change amount greater than or equal to a predetermined reference value, the barrier function is reduced (for example, the permeability of the substance from the outside to the inside of the skin) Can be evaluated). On the other hand, when the amount of moisture in the skin after contact with the test substance or article changes with an amount of change smaller than the reference value, it can be evaluated that the barrier function has not changed.
本発明では、上述のような皮膚の水分量又は水分分布と、バリア機能との関連性に基づいて、非侵襲的に測定した皮膚の水分量又は水分分布の変化量から、被験物質又は物品が、皮膚のバリア機能に及ぼす影響を評価する。具体的には、被験物質又は物品の接触後の皮膚の水分量が、所定の基準値以上の変化量で増加した場合、バリア機能を低下させる(例えば、皮膚外部から内部への物質の浸透性を亢進させる)と評価することができる。一方、被験物質又は物品の接触後の皮膚の水分量が、基準値より小さい変化量で変化した場合、バリア機能は変化してないと評価することができる。 As shown in Examples described later, the skin moisture amount or moisture distribution measured by Raman spectroscopy, skin capacitance measuring means, or the like has a strong relationship with the skin barrier function. Specifically, when the amount of change in the amount of moisture in the skin increases by a certain value or more due to contact with the test substance or article, the permeability of substances such as methyl nicotinate to the skin is enhanced.
In the present invention, based on the relationship between the moisture content or moisture distribution of the skin as described above and the barrier function, the test substance or article is obtained from the amount of change in the moisture content or moisture distribution measured noninvasively. Evaluate the effects on skin barrier function. Specifically, when the amount of moisture in the skin after contact with the test substance or article increases with a change amount greater than or equal to a predetermined reference value, the barrier function is reduced (for example, the permeability of the substance from the outside to the inside of the skin) Can be evaluated). On the other hand, when the amount of moisture in the skin after contact with the test substance or article changes with an amount of change smaller than the reference value, it can be evaluated that the barrier function has not changed.
例えば、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚静電容量測定装置(静電容量から計算された水分量の相対値(A.U.)を算出する装置)を用いて測定した、被験物質又は物品の接触前後の皮膚の水分量の変化量の基準値を、8(A.U.)、好ましくは12(A.U.)、より好ましくは16(A.U.)に設定する。そして、実際に測定した被験物質又は物品の接触後の皮膚の水分量が、前記基準値以上増加した場合に、皮膚外部から内部への物質の浸透性が亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。これに対して、水分量の変化量が前記基準値より小さい場合、被験物質又は物品の接触によってもバリア機能は変化していないと評価できる。
For example, the skin capacitance measuring device (calculating the relative value (AU) of moisture calculated from the capacitance) for the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes The reference value for the amount of change in the moisture content of the skin before and after contact with the test substance or article is set to 8 (AU), preferably 12 (AU), more preferably 16 (AU). To do. Then, when the amount of moisture in the skin after contact with the test substance or article actually measured increases more than the reference value, the permeability of the substance from the outside to the inside of the skin is enhanced, and contact with the test substance or article It can be evaluated that the barrier function is lowered. On the other hand, when the amount of change in the amount of water is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
加えて、被験物質又は物品の接触前後の皮膚の水分分布を比較し、初期の角層厚を基準として、被験物質又は物品の接触により初期の角層厚よりも深いところの皮膚の水分量が増加した場合、皮膚外部から内部への物質の浸透性を亢進しやすくなり、バリア機能は低下したと評価できる。
具体的には、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、実際の皮膚表面から所定の深さまで(好ましくは皮膚表面からの深さ40μmの深さまで、より好ましくは皮膚表面からの深さ35μmの深さまで、より好ましくは皮膚表面からの深さ20μmの深さまで)における、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合に、皮膚外部から内部への物質の浸透性が亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。 In addition, the moisture distribution of the skin before and after contact with the test substance or article is compared. Based on the initial stratum corneum thickness, the amount of skin moisture deeper than the initial stratum corneum due to the test substance or article contact When it increases, it becomes easy to enhance the permeability of the substance from the outside to the inside of the skin, and it can be evaluated that the barrier function is lowered.
Specifically, the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is the target, from the actual skin surface to a predetermined depth (preferably a depth of 40 μm from the skin surface). And more preferably to a depth of 35 μm from the skin surface, more preferably to a depth of 20 μm from the skin surface), the moisture distribution of the skin before and after contact with the test substance or article is measured by Raman spectroscopy. To do. Then, the moisture distribution in the skin before and after contact with the test substance or article is compared, and when the skin moisture content in the deep part is deeper than the initial stratum corneum thickness due to contact with the test substance or article, It can be evaluated that the permeability of the substance is enhanced and the barrier function is lowered by contact with the test substance or article.
具体的には、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、実際の皮膚表面から所定の深さまで(好ましくは皮膚表面からの深さ40μmの深さまで、より好ましくは皮膚表面からの深さ35μmの深さまで、より好ましくは皮膚表面からの深さ20μmの深さまで)における、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合に、皮膚外部から内部への物質の浸透性が亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。 In addition, the moisture distribution of the skin before and after contact with the test substance or article is compared. Based on the initial stratum corneum thickness, the amount of skin moisture deeper than the initial stratum corneum due to the test substance or article contact When it increases, it becomes easy to enhance the permeability of the substance from the outside to the inside of the skin, and it can be evaluated that the barrier function is lowered.
Specifically, the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is the target, from the actual skin surface to a predetermined depth (preferably a depth of 40 μm from the skin surface). And more preferably to a depth of 35 μm from the skin surface, more preferably to a depth of 20 μm from the skin surface), the moisture distribution of the skin before and after contact with the test substance or article is measured by Raman spectroscopy. To do. Then, the moisture distribution in the skin before and after contact with the test substance or article is compared, and when the skin moisture content in the deep part is deeper than the initial stratum corneum thickness due to contact with the test substance or article, It can be evaluated that the permeability of the substance is enhanced and the barrier function is lowered by contact with the test substance or article.
また、被験物質又は物品の接触前後の皮膚表面の水分量を測定し、被験物質又は物品の接触による、皮膚表面の水分量の変化量からも、バリア機能に及ぼす影響を評価できる。
具体的には、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚表面における、被験物質又は物品の接触前後の皮膚の水分量をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分量を比較する。本発明においては例えば、ラマン分光法より得られた皮膚表面水分量の被験物質又は物品の接触前後の変化量の基準値を2.0倍、より好ましくは2.5倍、さらに好ましくは3.0倍に設定する。そして、実際に測定した被験物質又は物品の接触後の、皮膚の表面水分量が前記基準値以上に増加した場合に、皮膚外部から内部への物質の浸透性が亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。これに対して、皮膚の表面水分量の変化量が前記基準値より小さい場合、被験物質又は物品の接触によってもバリア機能は変化していないと評価できる。 In addition, the moisture content on the skin surface before and after contact with the test substance or article can be measured, and the influence on the barrier function can be evaluated from the amount of change in the moisture content on the skin surface due to contact with the test substance or article.
Specifically, the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is targeted, and the moisture content of the skin before and after contact with the test substance or article on the skin surface is determined by Raman spectroscopy. Measure and compare the moisture content of the skin before and after contact with the test substance or article. In the present invention, for example, the reference value of the amount of change in the moisture content on the skin surface obtained by Raman spectroscopy before and after contact with the test substance or article is 2.0 times, more preferably 2.5 times, and even more preferably 3. Set to 0 times. When the surface moisture content of the skin after contact with the actually measured test substance or article increases above the reference value, the permeability of the substance from the outside of the skin to the inside increases, and the test substance or article It can be evaluated that the barrier function is lowered by the contact. On the other hand, when the amount of change in the surface moisture content of the skin is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
具体的には、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚表面における、被験物質又は物品の接触前後の皮膚の水分量をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分量を比較する。本発明においては例えば、ラマン分光法より得られた皮膚表面水分量の被験物質又は物品の接触前後の変化量の基準値を2.0倍、より好ましくは2.5倍、さらに好ましくは3.0倍に設定する。そして、実際に測定した被験物質又は物品の接触後の、皮膚の表面水分量が前記基準値以上に増加した場合に、皮膚外部から内部への物質の浸透性が亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。これに対して、皮膚の表面水分量の変化量が前記基準値より小さい場合、被験物質又は物品の接触によってもバリア機能は変化していないと評価できる。 In addition, the moisture content on the skin surface before and after contact with the test substance or article can be measured, and the influence on the barrier function can be evaluated from the amount of change in the moisture content on the skin surface due to contact with the test substance or article.
Specifically, the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is targeted, and the moisture content of the skin before and after contact with the test substance or article on the skin surface is determined by Raman spectroscopy. Measure and compare the moisture content of the skin before and after contact with the test substance or article. In the present invention, for example, the reference value of the amount of change in the moisture content on the skin surface obtained by Raman spectroscopy before and after contact with the test substance or article is 2.0 times, more preferably 2.5 times, and even more preferably 3. Set to 0 times. When the surface moisture content of the skin after contact with the actually measured test substance or article increases above the reference value, the permeability of the substance from the outside of the skin to the inside increases, and the test substance or article It can be evaluated that the barrier function is lowered by the contact. On the other hand, when the amount of change in the surface moisture content of the skin is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
また、被験物質又は物品の接触前後の初期の角層厚に相当する深さの水分量を測定し、被験物質又は物品の接触による、初期の角層厚に相当する深さの水分量の変化量からも、バリア機能に及ぼす影響を評価できる。
具体的には、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、初期の角層厚に相当する部位における、被験物質又は物品の接触前後の皮膚の水分量をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分量を比較する。本発明においては例えば、ラマン分光法より得られた初期の角層厚に相当する部位の水分量の被験物質又は物品の接触前後の変化量の基準値を8%、好ましくは11%、さらに好ましくは14%に設定する。そして、実際に測定した被験物質又は物品の接触後の、初期の角層厚に相当する部位の水分量が前記基準値以上増加した場合に、皮膚外部から内部への物質の浸透性を亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。これに対して、初期の角層厚に相当する部位の水分量の変化量が前記基準値より小さい場合、被験物質又は物品の接触によってもバリア機能は変化していないと評価できる。 Also, the moisture content at the depth corresponding to the initial stratum corneum thickness before and after contact with the test substance or article is measured, and the change in the moisture content at the depth corresponding to the initial stratum corneum due to the contact with the test substance or article. The effect on the barrier function can also be evaluated from the amount.
Specifically, the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is targeted, and the skin before and after contact with the test substance or article at the site corresponding to the initial stratum corneum thickness The moisture content is measured by Raman spectroscopy, and the moisture content of the skin before and after contact with the test substance or article is compared. In the present invention, for example, the reference value of the amount of change before and after the contact of the test substance or article with the water content at the site corresponding to the initial stratum corneum thickness obtained by Raman spectroscopy is 8%, preferably 11%, more preferably Is set to 14%. Then, when the amount of water at the site corresponding to the initial stratum corneum thickness after contact with the actually measured test substance or article increases more than the reference value, the permeability of the substance from outside the skin to the inside is enhanced. It can be evaluated that the barrier function has decreased due to contact with the test substance or article. On the other hand, when the amount of change in the amount of water at the site corresponding to the initial stratum corneum thickness is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
具体的には、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、初期の角層厚に相当する部位における、被験物質又は物品の接触前後の皮膚の水分量をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分量を比較する。本発明においては例えば、ラマン分光法より得られた初期の角層厚に相当する部位の水分量の被験物質又は物品の接触前後の変化量の基準値を8%、好ましくは11%、さらに好ましくは14%に設定する。そして、実際に測定した被験物質又は物品の接触後の、初期の角層厚に相当する部位の水分量が前記基準値以上増加した場合に、皮膚外部から内部への物質の浸透性を亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。これに対して、初期の角層厚に相当する部位の水分量の変化量が前記基準値より小さい場合、被験物質又は物品の接触によってもバリア機能は変化していないと評価できる。 Also, the moisture content at the depth corresponding to the initial stratum corneum thickness before and after contact with the test substance or article is measured, and the change in the moisture content at the depth corresponding to the initial stratum corneum due to the contact with the test substance or article. The effect on the barrier function can also be evaluated from the amount.
Specifically, the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is targeted, and the skin before and after contact with the test substance or article at the site corresponding to the initial stratum corneum thickness The moisture content is measured by Raman spectroscopy, and the moisture content of the skin before and after contact with the test substance or article is compared. In the present invention, for example, the reference value of the amount of change before and after the contact of the test substance or article with the water content at the site corresponding to the initial stratum corneum thickness obtained by Raman spectroscopy is 8%, preferably 11%, more preferably Is set to 14%. Then, when the amount of water at the site corresponding to the initial stratum corneum thickness after contact with the actually measured test substance or article increases more than the reference value, the permeability of the substance from outside the skin to the inside is enhanced. It can be evaluated that the barrier function has decreased due to contact with the test substance or article. On the other hand, when the amount of change in the amount of water at the site corresponding to the initial stratum corneum thickness is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article.
あるいは、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、ラマン分光法により測定した、皮膚表面からの所定の深さまで(好ましくは皮膚表面からの深さ2~40μmの深さまで、より好ましくは皮膚表面からの深さ2~35μmの深さまで、より好ましくは皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで)における、被験物質又は物品の接触前後の皮膚の水分量の曲線下面積(Area under the curve of water profile)を求め、被験物質又は物品の接触前後の曲線下面積を比較する。本発明においては例えば、ラマン分光法より得られた被験物質又は物品の接触前後の曲線下面積の変化量の基準値を、1.4倍、好ましくは1.6倍、さらに好ましくは1.8倍に設定する。そして、実際に測定した被験物質又は物品の接触後の、皮膚表面から所定の深さまでにおける曲線下面積が前記基準値以上に増加した場合に、皮膚外部から内部への物質の浸透性が亢進し、被験物質又は物品の接触によりバリア機能は低下したと評価することができる。これに対して、曲線下面積の変化量が前記基準値より小さい場合、被験物質又は物品の接触によってもバリア機能は変化していないと評価できる。なお本明細書において「面積下曲線」は、角層における水分量を示すものであって、深さごとの水分量を深さ方向に積分することによって求められる。
Alternatively, the skin inside the forearm of a healthy person after washing and acclimatization for at least 20 minutes is used as a target, and is measured by Raman spectroscopy to a predetermined depth from the skin surface (preferably a depth from the skin surface). To a depth of 2 to 40 μm, more preferably to a depth of 2 to 35 μm from the skin surface, more preferably to a depth of 2 to 20 μm from the skin surface, more preferably to a depth of 9 to The area under the curve of the moisture content of the skin before and after contact with the test substance or article at a depth of 15 μm, more preferably up to a depth of about 13 μm from the skin surface, and (AreaAunder the curve of water profile) Compare the area under the curve before and after contact of the test substance or article. In the present invention, for example, the reference value of the amount of change in the area under the curve before and after contact of the test substance or article obtained by Raman spectroscopy is 1.4 times, preferably 1.6 times, more preferably 1.8. Set to double. Then, when the area under the curve from the skin surface to the predetermined depth after contact with the actually measured test substance or article increases beyond the reference value, the permeability of the substance from outside the skin to the inside increases. It can be evaluated that the barrier function has decreased due to contact with the test substance or article. On the other hand, when the amount of change in the area under the curve is smaller than the reference value, it can be evaluated that the barrier function is not changed by contact with the test substance or article. In the present specification, the “lower area curve” indicates the moisture content in the stratum corneum, and is obtained by integrating the moisture content for each depth in the depth direction.
ラマン分光法による皮膚の水分量の測定は、特開2012-50739号公報に記載の方法を参照して行うことができる。本方法は、皮膚試料をタンパク質、水、および脂質の三成分系で近似し、脂質由来のラマンスペクトルの寄与分を、皮膚の実測ラマンスペクトルから差し引いた補正ラマンスペクトルを作成し、タンパク質由来のCH伸縮信号強度に対する水のOH伸縮振動由来の信号強度の比率に基づき皮膚水分量を測定する方法である。以下、ラマン分光法による皮膚の水分量の測定方法について詳細に説明する。しかし、本発明はこれに制限するものではない。
Measurement of the moisture content of the skin by Raman spectroscopy can be performed with reference to the method described in JP2012-50739A. This method approximates a skin sample with a ternary system of protein, water, and lipid, creates a corrected Raman spectrum by subtracting the contribution from the lipid-derived Raman spectrum from the measured Raman spectrum of the skin, and generates a protein-derived CH This is a method of measuring the amount of skin moisture based on the ratio of the signal intensity derived from the OH stretching vibration of water to the stretching signal intensity. Hereinafter, a method for measuring the moisture content of the skin by Raman spectroscopy will be described in detail. However, the present invention is not limited to this.
まず、ベースラインの蛍光の補正をおこなう。測定した皮膚のラマンスペクトルから蛍光によるベースラインのゆがみを排除するため、2000~2800cm-1および3800~4200cm-1の波数領域のデータより、最小二乗法をもちいた三次関数近似に基づきベースラインを予測し、これを差し引く。
蛍光の影響を排除した補正後の皮膚のスペクトルを、特開2012-50739号公報記載の式(4)を用いて脱脂乾燥角層、モデル皮脂、モデル細胞間脂質及び水のラマンスペクトル4つの重ねあわせで近似する。すなわち、最小二乗法により前記式(4)における係数C1~C4を決定する。このときC2ISEB(ω)が皮脂の寄与分、C3ICER(ω)が細胞間脂質の寄与分となる。以下に示すモデル脂質のラマンスペクトルにおけるモデル脂質の寄与量を最小二乗法で見積もる。
そして、皮脂の寄与分及び細胞間脂質の寄与分を差し引いた特開2012-50739号公報記載の式(5)で表される補正ラマンスペクトルを得る。次に、式(5)で表される補正ラマンスペクトルにおけるCH伸縮信号(波数領域:2800~3030cm-1)のピーク面積に対するOH伸縮振動のピーク面積(波数領域:3100~3750cm-1)の比率を算出する。そして、ラマンスペクトルにおけるタンパク質のCH伸縮振動由来の信号強度に対する水のOH伸縮振動由来の信号強度の比率と、該脱脂皮膚試料に含まれる水分量とタンパク質量の比とをプロットして得られる検量線(例えば、特開2012-50739号公報の図5参照)に基づいて皮膚の水分量(質量%)を算出する。 First, the baseline fluorescence is corrected. Since the Raman spectrum of the measured skin to eliminate the distortion of baseline by fluorescence, from the data of the wavenumber region of 2000 ~ 2800 cm -1 and 3800 ~ 4200cm -1, a baseline based on cubic function approximation using the least square method Predict and subtract this.
The corrected skin spectrum excluding the influence of fluorescence is superimposed on four Raman spectra of degreased and dried stratum corneum, model sebum, model intercellular lipid and water using equation (4) described in JP2012-50739A. Approximate together. That is, the coefficients C 1 to C 4 in the equation (4) are determined by the least square method. At this time, C 2 I SEB (ω) is the contribution of sebum, and C 3 I CER (ω) is the contribution of the intercellular lipid. The contribution amount of the model lipid in the Raman spectrum of the model lipid shown below is estimated by the least square method.
Then, a corrected Raman spectrum represented by the formula (5) described in JP 2012-50739 A is obtained by subtracting the contribution of sebum and the contribution of intercellular lipids. Then, CH stretch signal (wavenumber region: 2800 ~ 3030cm -1) in the correction Raman spectra of the formula (5) OH stretching vibration peak area (wavenumber region: 3100 ~ 3750cm -1) to the peak area ratio of Is calculated. A calibration obtained by plotting the ratio of the signal intensity derived from the OH stretching vibration of water to the signal intensity derived from the CH stretching vibration of protein in the Raman spectrum and the ratio of the amount of water and the amount of protein contained in the defatted skin sample. The moisture content (mass%) of the skin is calculated based on a line (for example, see FIG. 5 of JP2012-50739A).
蛍光の影響を排除した補正後の皮膚のスペクトルを、特開2012-50739号公報記載の式(4)を用いて脱脂乾燥角層、モデル皮脂、モデル細胞間脂質及び水のラマンスペクトル4つの重ねあわせで近似する。すなわち、最小二乗法により前記式(4)における係数C1~C4を決定する。このときC2ISEB(ω)が皮脂の寄与分、C3ICER(ω)が細胞間脂質の寄与分となる。以下に示すモデル脂質のラマンスペクトルにおけるモデル脂質の寄与量を最小二乗法で見積もる。
そして、皮脂の寄与分及び細胞間脂質の寄与分を差し引いた特開2012-50739号公報記載の式(5)で表される補正ラマンスペクトルを得る。次に、式(5)で表される補正ラマンスペクトルにおけるCH伸縮信号(波数領域:2800~3030cm-1)のピーク面積に対するOH伸縮振動のピーク面積(波数領域:3100~3750cm-1)の比率を算出する。そして、ラマンスペクトルにおけるタンパク質のCH伸縮振動由来の信号強度に対する水のOH伸縮振動由来の信号強度の比率と、該脱脂皮膚試料に含まれる水分量とタンパク質量の比とをプロットして得られる検量線(例えば、特開2012-50739号公報の図5参照)に基づいて皮膚の水分量(質量%)を算出する。 First, the baseline fluorescence is corrected. Since the Raman spectrum of the measured skin to eliminate the distortion of baseline by fluorescence, from the data of the wavenumber region of 2000 ~ 2800 cm -1 and 3800 ~ 4200cm -1, a baseline based on cubic function approximation using the least square method Predict and subtract this.
The corrected skin spectrum excluding the influence of fluorescence is superimposed on four Raman spectra of degreased and dried stratum corneum, model sebum, model intercellular lipid and water using equation (4) described in JP2012-50739A. Approximate together. That is, the coefficients C 1 to C 4 in the equation (4) are determined by the least square method. At this time, C 2 I SEB (ω) is the contribution of sebum, and C 3 I CER (ω) is the contribution of the intercellular lipid. The contribution amount of the model lipid in the Raman spectrum of the model lipid shown below is estimated by the least square method.
Then, a corrected Raman spectrum represented by the formula (5) described in JP 2012-50739 A is obtained by subtracting the contribution of sebum and the contribution of intercellular lipids. Then, CH stretch signal (wavenumber region: 2800 ~ 3030cm -1) in the correction Raman spectra of the formula (5) OH stretching vibration peak area (wavenumber region: 3100 ~ 3750cm -1) to the peak area ratio of Is calculated. A calibration obtained by plotting the ratio of the signal intensity derived from the OH stretching vibration of water to the signal intensity derived from the CH stretching vibration of protein in the Raman spectrum and the ratio of the amount of water and the amount of protein contained in the defatted skin sample. The moisture content (mass%) of the skin is calculated based on a line (for example, see FIG. 5 of JP2012-50739A).
前述の脱脂乾燥角層、モデル皮脂、モデル細胞間脂質の調製方法の詳細を示す。しかし、本発明はこれに制限するものではない。
脱脂乾燥角層
健常男性(40代)のかかとより角層片をナイフで切除後、クロロホルムーメタノール(1:1)に一昼夜浸漬する。その後角層片試料を取り出し、自然乾燥させることで、脂質等の油溶性成分と、アミノ酸などの水溶性成分を除去した脱脂乾燥角層を調製する。
モデル皮脂
下記成分を混合し、モデル皮脂を調製する。
オレイン酸(関東化学製):17.4質量%
トリオレイン酸(関東化学製):43.4質量%
オレイン酸デシル(和光純薬製):26.5質量%
スクワレン(関東化学製):12.7質量%
モデル細胞間脂質
シグマ社製non-hydroxy fatty cermaideをモデル細胞間脂質とする。 The detail of the preparation method of the above-mentioned degreasing dry stratum corneum, model sebum, and model intercellular lipid is shown. However, the present invention is not limited to this.
A horny layer piece is excised with a knife from the heel of a degreased and dried horny layer healthy male (40's), and then immersed in chloroform-methanol (1: 1) overnight. Thereafter, a sample of the stratum corneum is taken out and naturally dried to prepare a degreased and dried stratum corneum from which oil-soluble components such as lipids and water-soluble components such as amino acids have been removed.
Model sebum The following components are mixed to prepare model sebum.
Oleic acid (manufactured by Kanto Chemical): 17.4% by mass
Trioleic acid (manufactured by Kanto Chemical): 43.4% by mass
Decyl oleate (Wako Pure Chemical Industries): 26.5% by mass
Squalene (manufactured by Kanto Chemical): 12.7% by mass
Model intercellular lipid Sigma's non-hydroxy fatty cermaide is used as model intercellular lipid.
脱脂乾燥角層
健常男性(40代)のかかとより角層片をナイフで切除後、クロロホルムーメタノール(1:1)に一昼夜浸漬する。その後角層片試料を取り出し、自然乾燥させることで、脂質等の油溶性成分と、アミノ酸などの水溶性成分を除去した脱脂乾燥角層を調製する。
モデル皮脂
下記成分を混合し、モデル皮脂を調製する。
オレイン酸(関東化学製):17.4質量%
トリオレイン酸(関東化学製):43.4質量%
オレイン酸デシル(和光純薬製):26.5質量%
スクワレン(関東化学製):12.7質量%
モデル細胞間脂質
シグマ社製non-hydroxy fatty cermaideをモデル細胞間脂質とする。 The detail of the preparation method of the above-mentioned degreasing dry stratum corneum, model sebum, and model intercellular lipid is shown. However, the present invention is not limited to this.
A horny layer piece is excised with a knife from the heel of a degreased and dried horny layer healthy male (40's), and then immersed in chloroform-methanol (1: 1) overnight. Thereafter, a sample of the stratum corneum is taken out and naturally dried to prepare a degreased and dried stratum corneum from which oil-soluble components such as lipids and water-soluble components such as amino acids have been removed.
Model sebum The following components are mixed to prepare model sebum.
Oleic acid (manufactured by Kanto Chemical): 17.4% by mass
Trioleic acid (manufactured by Kanto Chemical): 43.4% by mass
Decyl oleate (Wako Pure Chemical Industries): 26.5% by mass
Squalene (manufactured by Kanto Chemical): 12.7% by mass
Model intercellular lipid Sigma's non-hydroxy fatty cermaide is used as model intercellular lipid.
本発明で用いる物品としては、例えば、吸収性物品(脱脂綿、生理用ナプキン、紙おむつ、布おむつ、トレーニングパンツ、成人用おむつ、汗止めパット、尿取りパット、産褥パット、パンティーライナー、吸収パットなど)、蒸気温熱具、創傷保護材(絆創膏、ガーゼ、包帯など)、マスク、ゴム手袋などが挙げられる。
また、前記被験物質としては、水若しくは水蒸気、化粧料、保湿剤若しくは外用剤、蒸気温熱具に含まれる有効成分や薬効成分、生理食塩水、リン酸緩衝生理食塩水、及び生体由来の分泌物や***物である尿、汗、経血、便などが挙げられる。なお、本発明において、擬似尿、疑似汗、擬似経血、擬似便などは生体由来の分泌物や***物に含まれる。
さらに、皮膚への被験物質又は物品の接触時間も適宜設定することができる。例えば、前記被験物質又は物品をヒトの皮膚に接触させる時間は、1時間以上6時間以下、好ましくは2時間以上4時間以下、より好ましくは2.5時間以上3.5時間以下、さらに好ましくは3時間程度である。 Examples of articles used in the present invention include absorbent articles (absorbent cotton, sanitary napkins, paper diapers, cloth diapers, training pants, adult diapers, sweat pads, urine pads, puerperium pads, panty liners, absorbent pads, etc.) , Steam heaters, wound protection materials (such as bandages, gauze, bandages, etc.), masks, and rubber gloves.
Examples of the test substance include water or water vapor, cosmetics, humectants or external preparations, active ingredients and medicinal ingredients contained in steam heating devices, physiological saline, phosphate buffered saline, and biologically derived secretions. And urine such as urine, sweat, menstrual blood, and feces. In the present invention, simulated urine, simulated sweat, simulated menstrual blood, simulated stool, etc. are included in living body-derived secretions and excretion.
Furthermore, the contact time of the test substance or article with the skin can also be set as appropriate. For example, the time for contacting the test substance or article with human skin is 1 hour or more and 6 hours or less, preferably 2 hours or more and 4 hours or less, more preferably 2.5 hours or more and 3.5 hours or less, and further preferably About 3 hours.
また、前記被験物質としては、水若しくは水蒸気、化粧料、保湿剤若しくは外用剤、蒸気温熱具に含まれる有効成分や薬効成分、生理食塩水、リン酸緩衝生理食塩水、及び生体由来の分泌物や***物である尿、汗、経血、便などが挙げられる。なお、本発明において、擬似尿、疑似汗、擬似経血、擬似便などは生体由来の分泌物や***物に含まれる。
さらに、皮膚への被験物質又は物品の接触時間も適宜設定することができる。例えば、前記被験物質又は物品をヒトの皮膚に接触させる時間は、1時間以上6時間以下、好ましくは2時間以上4時間以下、より好ましくは2.5時間以上3.5時間以下、さらに好ましくは3時間程度である。 Examples of articles used in the present invention include absorbent articles (absorbent cotton, sanitary napkins, paper diapers, cloth diapers, training pants, adult diapers, sweat pads, urine pads, puerperium pads, panty liners, absorbent pads, etc.) , Steam heaters, wound protection materials (such as bandages, gauze, bandages, etc.), masks, and rubber gloves.
Examples of the test substance include water or water vapor, cosmetics, humectants or external preparations, active ingredients and medicinal ingredients contained in steam heating devices, physiological saline, phosphate buffered saline, and biologically derived secretions. And urine such as urine, sweat, menstrual blood, and feces. In the present invention, simulated urine, simulated sweat, simulated menstrual blood, simulated stool, etc. are included in living body-derived secretions and excretion.
Furthermore, the contact time of the test substance or article with the skin can also be set as appropriate. For example, the time for contacting the test substance or article with human skin is 1 hour or more and 6 hours or less, preferably 2 hours or more and 4 hours or less, more preferably 2.5 hours or more and 3.5 hours or less, and further preferably About 3 hours.
本発明の評価方法は前述のように、皮膚水分量又は水分分布を非侵襲的に測定し、物品又は被験物質の接触による皮膚の水分量又は水分分布の変化量と皮膚のバリア機能との関連性に基づいて、測定した皮膚の水分量の変化量から皮膚のバリア機能、特に物質の浸透性を評価する。
As described above, the evaluation method of the present invention measures the skin moisture content or moisture distribution noninvasively, and relates the relationship between the amount of change in skin moisture content or moisture distribution due to contact with an article or test substance and the skin barrier function. Based on the sex, the barrier function of the skin, in particular the permeability of the substance, is evaluated from the measured amount of change in the skin water content.
また、前述の、被験物質又は物品の接触による皮膚の水分量又は水分分布の変化量とバリア機能との関連性に基づいて、非侵襲的に測定した皮膚の水分量又は水分分布の変化量から、角層の状態を評価することができる。
例えば、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚静電容量測定装置を用いて測定した、被験物質又は物品負荷前後の皮膚の水分量の変化量の基準値を、8(A.U.)、好ましくは12(A.U.)、より好ましくは16(A.U.)に設定する。そして、実際に測定した被験物質又は物品負荷後の皮膚の水分量が、前記基準値以上増加した場合に、角層のバリア機能が低下したと評価することができる。
加えて、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚表面から所定の深さまで(好ましくは皮膚表面からの深さ40μmの深さまで、より好ましくは皮膚表面からの深さ35μmの深さまで、より好ましくは皮膚表面からの深さ20μmの深さまで)における、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合に、角層のバリア機能は低下した、と評価できる。
また、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚表面からの深さ2μmにおける、被験物質又は物品の接触前後の皮膚の表面水分量をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の表面水分量を比較する。そして、得られた皮膚表面水分量の被験物質又は物品の接触前後の変化量の基準値を2.0倍、より好ましくは2.5倍、さらに好ましくは3.0倍に設定する。そして、実際に測定した被験物質又は物品の接触後の皮膚の表面水分量が、前記基準値以上に増加した場合に、角層のバリア機能は低下した、と評価できる。
また、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、初期の角層厚に相当する部位における、被験物質又は物品の接触前後の皮膚の水分量をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の水分量を比較する。そして、ラマン分光法より得られた初期の角層厚に相当する部位の水分量の被験物質又は物品の接触前後の変化量の基準値を8%、好ましくは11%、さらに好ましくは14%に設定する。そして、実際に測定した被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、前記基準値以上増加した場合に、バリア機能は低下したと評価できる。
あるいは、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、ラマン分光法により測定した、皮膚表面からの所定の深さまで(好ましくは皮膚表面からの深さ2~40μmの深さまで、より好ましくは皮膚表面からの深さ2~35μmの深さまで、より好ましくは皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで)における、皮膚の水分量の曲線下面積(Area under the curve of water profile)を求める。被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対する被験物質又は物品の一定時間接触後の皮膚の水分量の曲線下面積の変化量の基準値を、1.4倍、好ましくは1.6倍、さらに好ましくは1.8倍に設定する。そして、実際に測定した被験物質又は物品負荷後の、皮膚表面から所定の深さまでにおける皮膚の水分量の曲線下面積が前記基準値以上に増加した場合に、角層のバリア機能は低下したと評価できる。 In addition, based on the relationship between the amount of change in skin moisture or moisture distribution due to contact with the test substance or article and the barrier function, the amount of change in skin moisture or moisture distribution measured non-invasively is also described. The state of the stratum corneum can be evaluated.
For example, the amount of moisture in the skin before and after loading of the test substance or article, measured using a skin capacitance measuring device, for the skin inside the forearm of a healthy subject after washing the skin and acclimatizing for at least 20 minutes The reference value of the change amount is set to 8 (AU), preferably 12 (AU), more preferably 16 (AU). And it can be evaluated that the barrier function of the stratum corneum is lowered when the moisture content of the skin after loading the actually measured test substance or article increases by more than the reference value.
In addition, the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is the target, and from the skin surface to a predetermined depth (preferably to a depth of 40 μm from the skin surface, more preferably The water distribution of the skin before and after contact with the test substance or article at a depth of 35 μm from the skin surface, more preferably to a depth of 20 μm from the skin surface) is measured by Raman spectroscopy. Then, the moisture distribution in the skin before and after contact with the test substance or article is compared, and when the skin moisture content in the deeper part than the initial stratum corneum thickness is increased by contact with the test substance or article, the barrier function of the stratum corneum is It can be evaluated that it has decreased.
In addition, the skin moisture before and after contact with the test substance or article at a depth of 2 μm from the skin surface was examined for the skin inside the forearm of a healthy person after the skin was washed and acclimatized for at least 20 minutes. Measure by spectroscopy. Then, the surface moisture content of the skin before and after contact with the test substance or article is compared. Then, the reference value of the amount of change of the obtained skin surface moisture before and after contact with the test substance or article is set to 2.0 times, more preferably 2.5 times, and even more preferably 3.0 times. And it can be evaluated that the barrier function of the stratum corneum is reduced when the surface moisture content of the skin after contact with the test substance or article actually measured increases above the reference value.
In addition, the amount of moisture in the skin before and after contact with the test substance or article at the site corresponding to the initial stratum corneum thickness is measured for the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes. Measured by Raman spectroscopy. Then, the moisture content of the skin before and after contact with the test substance or article is compared. The reference value of the amount of change before and after the contact of the test substance or article with the moisture content at the site corresponding to the initial stratum corneum thickness obtained by Raman spectroscopy is 8%, preferably 11%, more preferably 14%. Set. And when the moisture content of the site | part corresponded to the initial stratum corneum thickness after contact with the test substance or goods actually measured increased more than the said reference value, it can be evaluated that the barrier function fell.
Alternatively, the skin inside the forearm of a healthy person after washing and acclimatization for at least 20 minutes is used as a target, and is measured by Raman spectroscopy to a predetermined depth from the skin surface (preferably a depth from the skin surface). To a depth of 2 to 40 μm, more preferably to a depth of 2 to 35 μm from the skin surface, more preferably to a depth of 2 to 20 μm from the skin surface, more preferably to a depth of 9 to The area under the curve of water profile is determined at a depth of 15 μm, more preferably a depth of about 13 μm from the skin surface. The reference value of the amount of change in the area under the curve of the moisture content of the skin after contact with the test substance or article for a certain period of time relative to the area under the curve of the moisture content of the skin before contact with the test substance or article is preferably 1.4 times, preferably It is set to 1.6 times, more preferably 1.8 times. And, when the area under the curve of the moisture content of the skin from the skin surface to the predetermined depth after the test substance or article load actually measured increases above the reference value, the barrier function of the stratum corneum is reduced. Can be evaluated.
例えば、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚静電容量測定装置を用いて測定した、被験物質又は物品負荷前後の皮膚の水分量の変化量の基準値を、8(A.U.)、好ましくは12(A.U.)、より好ましくは16(A.U.)に設定する。そして、実際に測定した被験物質又は物品負荷後の皮膚の水分量が、前記基準値以上増加した場合に、角層のバリア機能が低下したと評価することができる。
加えて、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚表面から所定の深さまで(好ましくは皮膚表面からの深さ40μmの深さまで、より好ましくは皮膚表面からの深さ35μmの深さまで、より好ましくは皮膚表面からの深さ20μmの深さまで)における、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合に、角層のバリア機能は低下した、と評価できる。
また、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、皮膚表面からの深さ2μmにおける、被験物質又は物品の接触前後の皮膚の表面水分量をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の表面水分量を比較する。そして、得られた皮膚表面水分量の被験物質又は物品の接触前後の変化量の基準値を2.0倍、より好ましくは2.5倍、さらに好ましくは3.0倍に設定する。そして、実際に測定した被験物質又は物品の接触後の皮膚の表面水分量が、前記基準値以上に増加した場合に、角層のバリア機能は低下した、と評価できる。
また、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、初期の角層厚に相当する部位における、被験物質又は物品の接触前後の皮膚の水分量をラマン分光法により測定する。そして、被験物質又は物品の接触前後の皮膚の水分量を比較する。そして、ラマン分光法より得られた初期の角層厚に相当する部位の水分量の被験物質又は物品の接触前後の変化量の基準値を8%、好ましくは11%、さらに好ましくは14%に設定する。そして、実際に測定した被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、前記基準値以上増加した場合に、バリア機能は低下したと評価できる。
あるいは、皮膚を洗浄し、最低20分間順化した後の健常者の前腕内側の皮膚を対象とし、ラマン分光法により測定した、皮膚表面からの所定の深さまで(好ましくは皮膚表面からの深さ2~40μmの深さまで、より好ましくは皮膚表面からの深さ2~35μmの深さまで、より好ましくは皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで)における、皮膚の水分量の曲線下面積(Area under the curve of water profile)を求める。被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対する被験物質又は物品の一定時間接触後の皮膚の水分量の曲線下面積の変化量の基準値を、1.4倍、好ましくは1.6倍、さらに好ましくは1.8倍に設定する。そして、実際に測定した被験物質又は物品負荷後の、皮膚表面から所定の深さまでにおける皮膚の水分量の曲線下面積が前記基準値以上に増加した場合に、角層のバリア機能は低下したと評価できる。 In addition, based on the relationship between the amount of change in skin moisture or moisture distribution due to contact with the test substance or article and the barrier function, the amount of change in skin moisture or moisture distribution measured non-invasively is also described. The state of the stratum corneum can be evaluated.
For example, the amount of moisture in the skin before and after loading of the test substance or article, measured using a skin capacitance measuring device, for the skin inside the forearm of a healthy subject after washing the skin and acclimatizing for at least 20 minutes The reference value of the change amount is set to 8 (AU), preferably 12 (AU), more preferably 16 (AU). And it can be evaluated that the barrier function of the stratum corneum is lowered when the moisture content of the skin after loading the actually measured test substance or article increases by more than the reference value.
In addition, the skin on the inner side of the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes is the target, and from the skin surface to a predetermined depth (preferably to a depth of 40 μm from the skin surface, more preferably The water distribution of the skin before and after contact with the test substance or article at a depth of 35 μm from the skin surface, more preferably to a depth of 20 μm from the skin surface) is measured by Raman spectroscopy. Then, the moisture distribution in the skin before and after contact with the test substance or article is compared, and when the skin moisture content in the deeper part than the initial stratum corneum thickness is increased by contact with the test substance or article, the barrier function of the stratum corneum is It can be evaluated that it has decreased.
In addition, the skin moisture before and after contact with the test substance or article at a depth of 2 μm from the skin surface was examined for the skin inside the forearm of a healthy person after the skin was washed and acclimatized for at least 20 minutes. Measure by spectroscopy. Then, the surface moisture content of the skin before and after contact with the test substance or article is compared. Then, the reference value of the amount of change of the obtained skin surface moisture before and after contact with the test substance or article is set to 2.0 times, more preferably 2.5 times, and even more preferably 3.0 times. And it can be evaluated that the barrier function of the stratum corneum is reduced when the surface moisture content of the skin after contact with the test substance or article actually measured increases above the reference value.
In addition, the amount of moisture in the skin before and after contact with the test substance or article at the site corresponding to the initial stratum corneum thickness is measured for the skin inside the forearm of a healthy person after washing the skin and acclimatizing for at least 20 minutes. Measured by Raman spectroscopy. Then, the moisture content of the skin before and after contact with the test substance or article is compared. The reference value of the amount of change before and after the contact of the test substance or article with the moisture content at the site corresponding to the initial stratum corneum thickness obtained by Raman spectroscopy is 8%, preferably 11%, more preferably 14%. Set. And when the moisture content of the site | part corresponded to the initial stratum corneum thickness after contact with the test substance or goods actually measured increased more than the said reference value, it can be evaluated that the barrier function fell.
Alternatively, the skin inside the forearm of a healthy person after washing and acclimatization for at least 20 minutes is used as a target, and is measured by Raman spectroscopy to a predetermined depth from the skin surface (preferably a depth from the skin surface). To a depth of 2 to 40 μm, more preferably to a depth of 2 to 35 μm from the skin surface, more preferably to a depth of 2 to 20 μm from the skin surface, more preferably to a depth of 9 to The area under the curve of water profile is determined at a depth of 15 μm, more preferably a depth of about 13 μm from the skin surface. The reference value of the amount of change in the area under the curve of the moisture content of the skin after contact with the test substance or article for a certain period of time relative to the area under the curve of the moisture content of the skin before contact with the test substance or article is preferably 1.4 times, preferably It is set to 1.6 times, more preferably 1.8 times. And, when the area under the curve of the moisture content of the skin from the skin surface to the predetermined depth after the test substance or article load actually measured increases above the reference value, the barrier function of the stratum corneum is reduced. Can be evaluated.
本発明の評価方法を利用して、吸収性物品(脱脂綿、生理用ナプキン、紙おむつ、布おむつ、トレーニングパンツ、成人用おむつ、汗止めパット、尿取りパット、産褥パット、パンティーライナー、吸収パットなど)、皮膚化粧料若しくは外用剤、創傷保護材(絆創膏、ガーゼ、包帯など)、マスク若しくはゴム手袋、蒸気温熱具などの製品の性能を評価することができる。
例えば、吸収性物品、創傷保護材、並びにマスク若しくはゴム手袋については、これらの着用前後の皮膚のバリア機能を本発明の評価方法を利用して評価することで、これらの製品の性能(例えば、水分(濡れ)の吸収性、水蒸気(蒸れ)の外部への発散性、皮膚トラブルの起こしやすさ)を、安全で簡便に短時間で評価できる。なお、吸収性物品の評価の際には、生理食塩水や擬似尿などを含ませた吸収性物品を評価に用いてもよい。
本発明により物品の着用や適用後の皮膚の水分量又は水分分布を評価すれば、ヒトの皮膚に本実施例で使用したニコチン酸メチルなどの物質を塗布する必要なく、物質の透過性を評価することができる。具体的には、吸収性物品、創傷保護材、並びにマスク若しくはゴム手袋について、現行品と改良品との比較の際に、装着前後の皮膚の水分量又は水分分布から物質透過性を推し量ることができる。このようにすれば、吸収性物品などの開発を効率よく行うことができる。
また、皮膚の水和状態を利用して、化粧料、外用剤などに含まれる有効成分を皮膚に浸透させることができることが報告されている(例えば、J. Invest. Dermatol., 1963, vol. 41, p. 307-311参照)。よって、化粧料、外用剤、蒸気温熱具などの皮膚への塗布、貼付又は着用前後の皮膚のバリア機能を本発明の評価方法を利用して評価することで、これらの製品に含まれる有効成分などの皮膚への透過性を、安全で簡便に短時間で評価できる。 Absorbent articles (absorbent cotton, sanitary napkins, disposable diapers, cloth diapers, training pants, adult diapers, sweat pads, urine pads, puerperium pads, panty liners, absorbent pads, etc.) using the evaluation method of the present invention It is possible to evaluate the performance of products such as skin cosmetics or external preparations, wound protection materials (such as bandages, gauze, bandages, etc.), masks or rubber gloves, and steam heaters.
For example, for absorbent articles, wound protection materials, and masks or rubber gloves, the performance of these products (e.g., by evaluating the barrier function of the skin before and after wearing using the evaluation method of the present invention) The ability to absorb moisture (wet), the ability of water vapor (steaming) to spread to the outside, and the likelihood of skin problems) can be evaluated safely and simply in a short time. In evaluating the absorbent article, an absorbent article containing physiological saline, simulated urine, or the like may be used for the evaluation.
If the moisture content or moisture distribution of the skin after the wearing or application of the article is evaluated according to the present invention, the permeability of the substance is evaluated without the need to apply a substance such as methyl nicotinate used in this example to human skin. can do. Specifically, for absorbent articles, wound protection materials, and masks or rubber gloves, the substance permeability can be estimated from the moisture content or moisture distribution of the skin before and after wearing when comparing the current product with the improved product. it can. If it does in this way, development of an absorptive article etc. can be performed efficiently.
In addition, it has been reported that active ingredients contained in cosmetics, external preparations and the like can be permeated into the skin using the hydration state of the skin (for example, J. Invest. Dermatol., 1963, vol. 41, p. 307-311). Therefore, the active ingredient contained in these products can be evaluated by using the evaluation method of the present invention to evaluate the barrier function of the skin before and after application, application, or wearing on the skin of cosmetics, external preparations, steam heaters, etc. The permeability to the skin such as can be evaluated safely and simply in a short time.
例えば、吸収性物品、創傷保護材、並びにマスク若しくはゴム手袋については、これらの着用前後の皮膚のバリア機能を本発明の評価方法を利用して評価することで、これらの製品の性能(例えば、水分(濡れ)の吸収性、水蒸気(蒸れ)の外部への発散性、皮膚トラブルの起こしやすさ)を、安全で簡便に短時間で評価できる。なお、吸収性物品の評価の際には、生理食塩水や擬似尿などを含ませた吸収性物品を評価に用いてもよい。
本発明により物品の着用や適用後の皮膚の水分量又は水分分布を評価すれば、ヒトの皮膚に本実施例で使用したニコチン酸メチルなどの物質を塗布する必要なく、物質の透過性を評価することができる。具体的には、吸収性物品、創傷保護材、並びにマスク若しくはゴム手袋について、現行品と改良品との比較の際に、装着前後の皮膚の水分量又は水分分布から物質透過性を推し量ることができる。このようにすれば、吸収性物品などの開発を効率よく行うことができる。
また、皮膚の水和状態を利用して、化粧料、外用剤などに含まれる有効成分を皮膚に浸透させることができることが報告されている(例えば、J. Invest. Dermatol., 1963, vol. 41, p. 307-311参照)。よって、化粧料、外用剤、蒸気温熱具などの皮膚への塗布、貼付又は着用前後の皮膚のバリア機能を本発明の評価方法を利用して評価することで、これらの製品に含まれる有効成分などの皮膚への透過性を、安全で簡便に短時間で評価できる。 Absorbent articles (absorbent cotton, sanitary napkins, disposable diapers, cloth diapers, training pants, adult diapers, sweat pads, urine pads, puerperium pads, panty liners, absorbent pads, etc.) using the evaluation method of the present invention It is possible to evaluate the performance of products such as skin cosmetics or external preparations, wound protection materials (such as bandages, gauze, bandages, etc.), masks or rubber gloves, and steam heaters.
For example, for absorbent articles, wound protection materials, and masks or rubber gloves, the performance of these products (e.g., by evaluating the barrier function of the skin before and after wearing using the evaluation method of the present invention) The ability to absorb moisture (wet), the ability of water vapor (steaming) to spread to the outside, and the likelihood of skin problems) can be evaluated safely and simply in a short time. In evaluating the absorbent article, an absorbent article containing physiological saline, simulated urine, or the like may be used for the evaluation.
If the moisture content or moisture distribution of the skin after the wearing or application of the article is evaluated according to the present invention, the permeability of the substance is evaluated without the need to apply a substance such as methyl nicotinate used in this example to human skin. can do. Specifically, for absorbent articles, wound protection materials, and masks or rubber gloves, the substance permeability can be estimated from the moisture content or moisture distribution of the skin before and after wearing when comparing the current product with the improved product. it can. If it does in this way, development of an absorptive article etc. can be performed efficiently.
In addition, it has been reported that active ingredients contained in cosmetics, external preparations and the like can be permeated into the skin using the hydration state of the skin (for example, J. Invest. Dermatol., 1963, vol. 41, p. 307-311). Therefore, the active ingredient contained in these products can be evaluated by using the evaluation method of the present invention to evaluate the barrier function of the skin before and after application, application, or wearing on the skin of cosmetics, external preparations, steam heaters, etc. The permeability to the skin such as can be evaluated safely and simply in a short time.
本発明の、被験物質又は物品が、皮膚のバリア機能に与える影響を評価するためのシステムは、被験物質又は物品の接触による皮膚の水分量又は水分分布の変化量と皮膚のバリア機能との関連性に関する情報を格納し、関連性に関する情報に基づき、被験物質又は物品の接触による皮膚の水分量又は水分分布の変化量から、前記影響を評価する。
本発明のシステムはいわゆるコンピュータであり、例えば、バスで相互に接続される、CPU(Central Processing Unit)、メモリ、入出力インタフェース等を有する。メモリは、RAM(Random Access Memory)、ROM(Read Only Memory)、ハードディスク、可搬型記憶媒体等である。入出力インタフェースは、表示装置や入力装置等のようなユーザインタフェース装置と接続される。入出力インタフェースは、ネットワークを介して他のコンピュータと通信を行う通信装置等と接続されてもよい。 The system for evaluating the influence of the test substance or article on the skin barrier function according to the present invention relates to the relationship between the amount of change in the moisture content or distribution of the skin due to the contact of the test substance or article and the skin barrier function. Information on sex is stored, and the influence is evaluated from the amount of change in the moisture content or moisture distribution of the skin due to contact with the test substance or article based on the information on the relevance.
The system of the present invention is a so-called computer and includes, for example, a CPU (Central Processing Unit), a memory, an input / output interface, and the like that are connected to each other via a bus. The memory is a RAM (Random Access Memory), a ROM (Read Only Memory), a hard disk, a portable storage medium, or the like. The input / output interface is connected to a user interface device such as a display device or an input device. The input / output interface may be connected to a communication device or the like that communicates with another computer via a network.
本発明のシステムはいわゆるコンピュータであり、例えば、バスで相互に接続される、CPU(Central Processing Unit)、メモリ、入出力インタフェース等を有する。メモリは、RAM(Random Access Memory)、ROM(Read Only Memory)、ハードディスク、可搬型記憶媒体等である。入出力インタフェースは、表示装置や入力装置等のようなユーザインタフェース装置と接続される。入出力インタフェースは、ネットワークを介して他のコンピュータと通信を行う通信装置等と接続されてもよい。 The system for evaluating the influence of the test substance or article on the skin barrier function according to the present invention relates to the relationship between the amount of change in the moisture content or distribution of the skin due to the contact of the test substance or article and the skin barrier function. Information on sex is stored, and the influence is evaluated from the amount of change in the moisture content or moisture distribution of the skin due to contact with the test substance or article based on the information on the relevance.
The system of the present invention is a so-called computer and includes, for example, a CPU (Central Processing Unit), a memory, an input / output interface, and the like that are connected to each other via a bus. The memory is a RAM (Random Access Memory), a ROM (Read Only Memory), a hard disk, a portable storage medium, or the like. The input / output interface is connected to a user interface device such as a display device or an input device. The input / output interface may be connected to a communication device or the like that communicates with another computer via a network.
本発明の評価方法及び評価システムは、ヒトの皮膚、好ましくは、被験物質又は物品を負荷していない状態で正常なバリア機能を有するヒト(健常者)の皮膚、を対象とすることが好ましい。
It is preferable that the evaluation method and the evaluation system of the present invention target human skin, preferably human (healthy person) skin having a normal barrier function in a state in which no test substance or article is loaded.
上述した実施形態に関し、本発明はさらに以下の評価方法、及び評価システムを開示する。
Regarding the embodiment described above, the present invention further discloses the following evaluation method and evaluation system.
<1>被験物質又は物品をヒトの皮膚に接触させ、
接触前後の皮膚の水分量又は水分分布の変化量を、基準値と比較し、
比較結果から、前記被験物質又は物品が、皮膚バリア機能に与える影響を評価する、
被験物質又は物品の評価方法であって、
水の負荷量を変えての皮膚への水負荷試験により測定された、皮膚の水分量又は水分分布と、皮膚のバリア機能とが関連付けられ、
皮膚の水分量又は水分分布と皮膚のバリア機能との関連性から、皮膚のバリア機能に変化が確認されるときの皮膚の水分量又は水分分布が、前記基準値として設定されたものである、
被験物質又は物品の評価方法。 <1> A test substance or article is brought into contact with human skin,
Compare the amount of moisture or moisture distribution in the skin before and after contact with the reference value,
From the comparison results, the effect of the test substance or article on the skin barrier function is evaluated.
A method for evaluating a test substance or article,
The water content or distribution of the skin, measured by a water load test on the skin with different water loads, is associated with the barrier function of the skin,
From the relationship between the moisture content or moisture distribution of the skin and the barrier function of the skin, the moisture content or moisture distribution of the skin when the change is confirmed in the skin barrier function is set as the reference value.
Evaluation method of test substance or article.
接触前後の皮膚の水分量又は水分分布の変化量を、基準値と比較し、
比較結果から、前記被験物質又は物品が、皮膚バリア機能に与える影響を評価する、
被験物質又は物品の評価方法であって、
水の負荷量を変えての皮膚への水負荷試験により測定された、皮膚の水分量又は水分分布と、皮膚のバリア機能とが関連付けられ、
皮膚の水分量又は水分分布と皮膚のバリア機能との関連性から、皮膚のバリア機能に変化が確認されるときの皮膚の水分量又は水分分布が、前記基準値として設定されたものである、
被験物質又は物品の評価方法。 <1> A test substance or article is brought into contact with human skin,
Compare the amount of moisture or moisture distribution in the skin before and after contact with the reference value,
From the comparison results, the effect of the test substance or article on the skin barrier function is evaluated.
A method for evaluating a test substance or article,
The water content or distribution of the skin, measured by a water load test on the skin with different water loads, is associated with the barrier function of the skin,
From the relationship between the moisture content or moisture distribution of the skin and the barrier function of the skin, the moisture content or moisture distribution of the skin when the change is confirmed in the skin barrier function is set as the reference value.
Evaluation method of test substance or article.
<2>皮膚のバリア機能の変化を確認するために、皮膚外部から内部への指標物質の浸透性がさらに測定され、指標物質の浸透性の結果から前記基準値が設定されたものである、前記<1>項に記載の方法。
<3>前記被験物質又は物品の接触後の皮膚水分量が、基準値以上の変化量で増加した場合、バリア機能が低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<1>又は<2>項に記載の方法。
<4>前記被験物質又は物品の接触後の皮膚水分量の変化量が、基準値より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<1>~<3>のいずれか1項に記載の方法。 <2> In order to confirm the change in the barrier function of the skin, the permeability of the indicator substance from the outside to the inside of the skin is further measured, and the reference value is set from the result of the permeability of the indicator substance. The method according to <1>.
<3> When the amount of skin moisture after contact with the test substance or article increases with a change amount greater than or equal to a reference value, the barrier function is reduced, preferably the permeability of the substance from the outside of the skin to the inside is increased, The method according to <1> or <2>, wherein
<4> When the amount of change in skin moisture after contact with the test substance or article is smaller than the reference value, the barrier function is not changed, preferably the permeability of the substance from the outside to the inside is enhanced. The method according to any one of <1> to <3>, wherein the method is evaluated as having no.
<3>前記被験物質又は物品の接触後の皮膚水分量が、基準値以上の変化量で増加した場合、バリア機能が低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<1>又は<2>項に記載の方法。
<4>前記被験物質又は物品の接触後の皮膚水分量の変化量が、基準値より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<1>~<3>のいずれか1項に記載の方法。 <2> In order to confirm the change in the barrier function of the skin, the permeability of the indicator substance from the outside to the inside of the skin is further measured, and the reference value is set from the result of the permeability of the indicator substance. The method according to <1>.
<3> When the amount of skin moisture after contact with the test substance or article increases with a change amount greater than or equal to a reference value, the barrier function is reduced, preferably the permeability of the substance from the outside of the skin to the inside is increased, The method according to <1> or <2>, wherein
<4> When the amount of change in skin moisture after contact with the test substance or article is smaller than the reference value, the barrier function is not changed, preferably the permeability of the substance from the outside to the inside is enhanced. The method according to any one of <1> to <3>, wherein the method is evaluated as having no.
<5>被験物質又は物品をヒトの皮膚に接触させた後のヒトの皮膚の深さごとの水分量を測定して、被験物質又は物品の接触後の水分分布を取得し、
被験物質又は物品の接触後の水分分布と基準水分分布とを比較することで、被験物質又は物品が、ヒトの皮膚のバリア機能に与える影響を評価する、被験物質又は物品の評価方法であって、
水又は水溶液のヒトの皮膚への負荷前に、ヒトの皮膚の深さ毎の水分量を測定して初期水分分布を取得し、
水又は水溶液のヒトの皮膚への負荷終了直後に、ヒトの皮膚の深さ毎の水分量の測定による水又は水溶液の負荷後の水分分布の取得と、指標物質を皮膚に適用して該指標物質の浸透度の測定を行い、
初期水分分布から水又は水溶液の負荷終了直後の水分分布への変化と、指標物質の浸透度から、指標物質の浸透度と水又は水溶液の負荷量との関連性を決定し、
前記関連性が変化したときのヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、前記基準水分分布とする、
被験物質又は物品の評価方法。 <5> Measure the moisture content for each depth of human skin after contacting the test substance or article with human skin, and obtain the moisture distribution after contact of the test substance or article,
A test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin by comparing the moisture distribution after contact of the test substance or article with a reference moisture distribution. ,
Before loading water or aqueous solution on human skin, measure the amount of water for each depth of human skin to obtain the initial moisture distribution,
Immediately after the end of loading of water or aqueous solution on human skin, acquisition of water distribution after loading water or aqueous solution by measuring the amount of water for each depth of human skin, and applying the indicator substance to the skin, the indicator Measure the permeability of the substance,
From the initial moisture distribution to the moisture distribution immediately after the end of the load of water or aqueous solution, and the permeability of the indicator substance, determine the relationship between the permeability of the indicator substance and the load of water or aqueous solution,
The water distribution after loading with water or an aqueous solution at every depth of the human skin when the relevance is changed as the reference water distribution,
Evaluation method of test substance or article.
被験物質又は物品の接触後の水分分布と基準水分分布とを比較することで、被験物質又は物品が、ヒトの皮膚のバリア機能に与える影響を評価する、被験物質又は物品の評価方法であって、
水又は水溶液のヒトの皮膚への負荷前に、ヒトの皮膚の深さ毎の水分量を測定して初期水分分布を取得し、
水又は水溶液のヒトの皮膚への負荷終了直後に、ヒトの皮膚の深さ毎の水分量の測定による水又は水溶液の負荷後の水分分布の取得と、指標物質を皮膚に適用して該指標物質の浸透度の測定を行い、
初期水分分布から水又は水溶液の負荷終了直後の水分分布への変化と、指標物質の浸透度から、指標物質の浸透度と水又は水溶液の負荷量との関連性を決定し、
前記関連性が変化したときのヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、前記基準水分分布とする、
被験物質又は物品の評価方法。 <5> Measure the moisture content for each depth of human skin after contacting the test substance or article with human skin, and obtain the moisture distribution after contact of the test substance or article,
A test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin by comparing the moisture distribution after contact of the test substance or article with a reference moisture distribution. ,
Before loading water or aqueous solution on human skin, measure the amount of water for each depth of human skin to obtain the initial moisture distribution,
Immediately after the end of loading of water or aqueous solution on human skin, acquisition of water distribution after loading water or aqueous solution by measuring the amount of water for each depth of human skin, and applying the indicator substance to the skin, the indicator Measure the permeability of the substance,
From the initial moisture distribution to the moisture distribution immediately after the end of the load of water or aqueous solution, and the permeability of the indicator substance, determine the relationship between the permeability of the indicator substance and the load of water or aqueous solution,
The water distribution after loading with water or an aqueous solution at every depth of the human skin when the relevance is changed as the reference water distribution,
Evaluation method of test substance or article.
<6>被験物質又は物品の接触前と接触後のヒトの皮膚の所定の深さ位置の水分量を測定し、接触前の水分量と接触後の水分量との差を取得し、
取得した水分量の差と、前記基準水分分布とに基づき、皮膚バリア機能に与える影響を評価する、前記<5>項に記載の評価方法。 <6> Measure the moisture content at a predetermined depth position of the human skin before and after contact with the test substance or article, and obtain the difference between the moisture content before contact and the moisture content after contact;
The evaluation method according to <5>, wherein the influence on the skin barrier function is evaluated based on the obtained difference in water amount and the reference water distribution.
取得した水分量の差と、前記基準水分分布とに基づき、皮膚バリア機能に与える影響を評価する、前記<5>項に記載の評価方法。 <6> Measure the moisture content at a predetermined depth position of the human skin before and after contact with the test substance or article, and obtain the difference between the moisture content before contact and the moisture content after contact;
The evaluation method according to <5>, wherein the influence on the skin barrier function is evaluated based on the obtained difference in water amount and the reference water distribution.
<7>前記指標物質が、ニコチン酸メチル、ニコチン酸ヘキシル、乳酸、黄色4号、赤色215号、カフェイン、ユビナール、カロテノイド、レチノール、並びに、皮膚保湿剤、皮膚外用剤、皮膚化粧料若しくは蒸気温熱具に含まれる有効成分若しくは薬効成分からなる群より選ばれる少なくとも1種である、前記<2>~<6>のいずれか1項に記載の方法。
<8>皮膚の水分量を皮膚静電容量測定手段により測定し解析する、前記<1>~<7>のいずれか1項に記載の方法。
<9>健常者の皮膚を評価対象とし、被験物質又は物品の接触後の皮膚の水分量を皮膚静電容量測定手段により測定し、被験物質又は物品の接触後の皮膚の水分量が、被験物質又は物品の接触前の皮膚の水分量に対して、8(A.U.)以上、好ましくは12(A.U.)以上、より好ましくは16(A.U.)以上、の変化量で増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<8>項に記載の方法。
<10>健常者の皮膚を評価対象とし、被験物質又は物品の接触後の皮膚の水分量を皮膚静電容量測定手段により測定し、被験物質又は物品の接触後の皮膚の水分量が、被験物質又は物品の接触前の皮膚の水分量に対して、16(A.U.)増加より小さい変化量、好ましくは12(A.U.)増加より小さい変化量、より好ましくは8(A.U.)増加より小さい変化量、の場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<8>項に記載の方法。
<11>皮膚表面からの深さ2~40μmの深さまで、好ましくは皮膚表面からの深さ2~35μmの深さまで、より好ましくは皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、の水分量又は水分分布をラマン分光法により測定し解析する、前記<1>~<7>のいずれか1項に記載の方法。
<12>健常者の皮膚を評価対象とし、皮膚表面から所定の深さまで、好ましくは皮膚表面からの深さ40μmの深さまで、より好ましくは皮膚表面からの深さ35μmの深さまで、より好ましくは皮膚表面からの深さ20μmの深さまで、における、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合、バリア機能が低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<13>健常者の皮膚を評価対象とし、ラマン分光法より得られた被験物質又は物品の接触後の皮膚表面水分量が、被験物質又は物品の接触前の皮膚表面水分量に対して、2.0倍以上、好ましくは2.5倍以上、より好ましくは3.0倍以上、に増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<14>健常者の皮膚を評価対象とし、ラマン分光法より得られた被験物質又は物品の接触後の皮膚表面水分量が、被験物質又は物品の接触前の皮膚表面水分量に対して、3.0倍増加より小さい場合、好ましくは2.5倍より小さい場合、より好ましくは2.0倍より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<11>項に記載の方法。
<15>健常者の皮膚を評価対象とし、ラマン分光法より得られた、被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、被験物質又は物品の接触前の初期の角層厚に相当する部位の水分量に対して、8%以上、好ましくは11%以上、より好ましくは14%以上、増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<16>健常者の皮膚を評価対象とし、ラマン分光法より得られた、被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、被験物質又は物品の接触前の初期の角層厚に相当する部位の水分量に対して、14%増加より小さい場合、好ましくは11%増加より小さい場合、さらに好ましくは8%増加より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<11>項に記載の方法。
<17>健常者の皮膚を評価対象とし、ラマン分光法より得られた、皮膚表面からの所定の深さまで、好ましくは皮膚表面からの深さ2~40μmの深さまで、より好ましくは皮膚表面からの深さ2~35μmの深さまで、より皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、における、被験物質又は物品の接触後の皮膚の水分量の曲線下面積を求め、被験物質又は物品の接触後の皮膚の水分量の曲線下面積が、被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対して1.4倍以上、好ましくは1.6倍以上、さらに好ましくは1.8倍以上、に増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<18>健常者の皮膚を評価対象とし、ラマン分光法より得られた、皮膚表面からの所定の深さまで、好ましくは皮膚表面からの深さ2~40μmの深さまで、より好ましくは皮膚表面からの深さ2~35μmの深さまで、より皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、における、被験物質又は物品の接触後の皮膚の水分量の曲線下面積を求め、被験物質又は物品の接触後の皮膚の水分量の曲線下面積が、被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対して1.8倍より小さい場合、好ましくは1.6倍より小さい場合、さらに好ましくは1.4倍より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<11>項に記載の方法。 <7> The indicator substance is methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4, red No. 215, caffeine, ubinal, carotenoid, retinol, and skin moisturizer, skin external preparation, skin cosmetic, or steam The method according to any one of <2> to <6>, wherein the method is at least one selected from the group consisting of an active ingredient or a medicinal ingredient contained in a heating tool.
<8> The method according to any one of <1> to <7>, wherein the moisture content of the skin is measured and analyzed by a skin capacitance measuring unit.
<9> Skin of healthy subject is evaluated, skin moisture after contact with test substance or article is measured by skin capacitance measuring means, and skin moisture after contact with test substance or article is tested. When the amount of change in the moisture content of the skin before contact with a substance or article increases by 8 (AU) or more, preferably 12 (AU) or more, more preferably 16 (AU) or more, the barrier function decreases. Preferably, the method according to <8> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is enhanced.
<10> Skin of healthy subject is evaluated, skin moisture after contact with test substance or article is measured by skin capacitance measuring means, and skin moisture after contact with test substance or article is tested A change of less than 16 (AU) increase, preferably a change of less than 12 (AU) increase, more preferably a change of less than 8 (AU) increase, relative to the amount of moisture in the skin before contact with the substance or article, In the case of <5>, the method according to <8>, wherein the barrier function is not changed, preferably the permeability of the substance from the outside to the inside of the skin is not enhanced.
<11> Up to a depth of 2 to 40 μm from the skin surface, preferably up to a depth of 2 to 35 μm from the skin surface, more preferably up to a depth of 2 to 20 μm from the skin surface, more preferably The water content or water distribution up to a depth of 9 to 15 μm from the skin surface, more preferably a depth of about 13 μm from the skin surface is measured and analyzed by Raman spectroscopy, <1> to < 7. The method according to any one of 7>.
<12> The skin of a healthy person is the subject of evaluation, up to a predetermined depth from the skin surface, preferably up to a depth of 40 μm from the skin surface, more preferably up to a depth of 35 μm from the skin surface, more preferably The skin moisture distribution before and after contact with the test substance or article up to a depth of 20 μm from the skin surface is measured by Raman spectroscopy, and the skin moisture distribution before and after contact with the test substance or article is compared. When the amount of moisture in the skin deeper than the initial stratum corneum thickness is increased by contact with the substance or article, it is evaluated that the barrier function is lowered, preferably the permeability of the substance from the outside to the inside of the skin is increased. The method according to <11> above.
<13> Skin surface moisture after contact with a test substance or article obtained by Raman spectroscopy is 2 with respect to the skin surface moisture before contact with the test substance or article. When it increases to 0.0 times or more, preferably 2.5 times or more, more preferably 3.0 times or more, the barrier function is lowered, preferably the permeability of the substance from the outside of the skin to the inside is increased, The method according to <11>, wherein the method is evaluated.
<14> The skin surface moisture after contact of the test substance or article obtained by Raman spectroscopy is 3 with respect to the skin surface moisture before contact of the test substance or article. If it is less than 0.0 times increase, preferably less than 2.5 times, more preferably less than 2.0 times, the barrier function is not changed, preferably the permeability of the substance from outside the skin to the inside is The method according to <11>, wherein the method is evaluated as not enhanced.
<15> The skin of a healthy person is the subject of evaluation, and the water content of the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the value before contact with the test substance or article. When the water content in the region corresponding to the initial stratum corneum thickness is increased by 8% or more, preferably 11% or more, more preferably 14% or more, the barrier function is lowered, preferably from the outside of the skin to the inside. The method according to <11>, wherein the permeability of the substance is evaluated to be enhanced.
<16> Using healthy skin as an evaluation target, the moisture content of the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the value before contact with the test substance or article. If the amount of water in the region corresponding to the initial stratum corneum thickness is less than 14% increase, preferably less than 11% increase, more preferably less than 8% increase, the barrier function is not changed, Preferably, the method according to <11> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is not enhanced.
<17> The skin of a healthy person is evaluated, and is obtained by Raman spectroscopy up to a predetermined depth from the skin surface, preferably a depth of 2 to 40 μm from the skin surface, more preferably from the skin surface. To a depth of 2 to 35 μm, more to a depth of 2 to 20 μm from the skin surface, more preferably to a depth of 9 to 15 μm from the skin surface, more preferably to a depth of 13 μm from the skin surface. The area under the curve of the moisture content of the skin after contact with the test substance or article up to a certain depth is obtained, and the area under the curve of the skin moisture amount after contact with the test substance or article is the contact of the test substance or article When increased to 1.4 times or more, preferably 1.6 times or more, more preferably 1.8 times or more with respect to the area under the curve of moisture content of the previous skin, the barrier function is lowered, preferably skin From outside to inside The method according to <11> above, wherein it is evaluated that the permeability of the substance to the substance is enhanced.
<18> The skin of a healthy person is evaluated, and obtained by Raman spectroscopy, to a predetermined depth from the skin surface, preferably to a depth of 2 to 40 μm from the skin surface, more preferably from the skin surface. To a depth of 2 to 35 μm, more to a depth of 2 to 20 μm from the skin surface, more preferably to a depth of 9 to 15 μm from the skin surface, more preferably to a depth of 13 μm from the skin surface. The area under the curve of the moisture content of the skin after contact with the test substance or article up to a certain depth is obtained, and the area under the curve of the skin moisture amount after contact with the test substance or article is the contact of the test substance or article If the area under the curve of water content of the previous skin is less than 1.8 times, preferably less than 1.6 times, more preferably less than 1.4 times, the barrier function is not changed, Like Alternatively, the method according to <11> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is not enhanced.
<8>皮膚の水分量を皮膚静電容量測定手段により測定し解析する、前記<1>~<7>のいずれか1項に記載の方法。
<9>健常者の皮膚を評価対象とし、被験物質又は物品の接触後の皮膚の水分量を皮膚静電容量測定手段により測定し、被験物質又は物品の接触後の皮膚の水分量が、被験物質又は物品の接触前の皮膚の水分量に対して、8(A.U.)以上、好ましくは12(A.U.)以上、より好ましくは16(A.U.)以上、の変化量で増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<8>項に記載の方法。
<10>健常者の皮膚を評価対象とし、被験物質又は物品の接触後の皮膚の水分量を皮膚静電容量測定手段により測定し、被験物質又は物品の接触後の皮膚の水分量が、被験物質又は物品の接触前の皮膚の水分量に対して、16(A.U.)増加より小さい変化量、好ましくは12(A.U.)増加より小さい変化量、より好ましくは8(A.U.)増加より小さい変化量、の場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<8>項に記載の方法。
<11>皮膚表面からの深さ2~40μmの深さまで、好ましくは皮膚表面からの深さ2~35μmの深さまで、より好ましくは皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、の水分量又は水分分布をラマン分光法により測定し解析する、前記<1>~<7>のいずれか1項に記載の方法。
<12>健常者の皮膚を評価対象とし、皮膚表面から所定の深さまで、好ましくは皮膚表面からの深さ40μmの深さまで、より好ましくは皮膚表面からの深さ35μmの深さまで、より好ましくは皮膚表面からの深さ20μmの深さまで、における、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合、バリア機能が低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<13>健常者の皮膚を評価対象とし、ラマン分光法より得られた被験物質又は物品の接触後の皮膚表面水分量が、被験物質又は物品の接触前の皮膚表面水分量に対して、2.0倍以上、好ましくは2.5倍以上、より好ましくは3.0倍以上、に増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<14>健常者の皮膚を評価対象とし、ラマン分光法より得られた被験物質又は物品の接触後の皮膚表面水分量が、被験物質又は物品の接触前の皮膚表面水分量に対して、3.0倍増加より小さい場合、好ましくは2.5倍より小さい場合、より好ましくは2.0倍より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<11>項に記載の方法。
<15>健常者の皮膚を評価対象とし、ラマン分光法より得られた、被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、被験物質又は物品の接触前の初期の角層厚に相当する部位の水分量に対して、8%以上、好ましくは11%以上、より好ましくは14%以上、増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<16>健常者の皮膚を評価対象とし、ラマン分光法より得られた、被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、被験物質又は物品の接触前の初期の角層厚に相当する部位の水分量に対して、14%増加より小さい場合、好ましくは11%増加より小さい場合、さらに好ましくは8%増加より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<11>項に記載の方法。
<17>健常者の皮膚を評価対象とし、ラマン分光法より得られた、皮膚表面からの所定の深さまで、好ましくは皮膚表面からの深さ2~40μmの深さまで、より好ましくは皮膚表面からの深さ2~35μmの深さまで、より皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、における、被験物質又は物品の接触後の皮膚の水分量の曲線下面積を求め、被験物質又は物品の接触後の皮膚の水分量の曲線下面積が、被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対して1.4倍以上、好ましくは1.6倍以上、さらに好ましくは1.8倍以上、に増加した場合、バリア機能は低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<11>項に記載の方法。
<18>健常者の皮膚を評価対象とし、ラマン分光法より得られた、皮膚表面からの所定の深さまで、好ましくは皮膚表面からの深さ2~40μmの深さまで、より好ましくは皮膚表面からの深さ2~35μmの深さまで、より皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、における、被験物質又は物品の接触後の皮膚の水分量の曲線下面積を求め、被験物質又は物品の接触後の皮膚の水分量の曲線下面積が、被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対して1.8倍より小さい場合、好ましくは1.6倍より小さい場合、さらに好ましくは1.4倍より小さい場合、バリア機能は変化していない、好ましくは皮膚外部から内部への物質の浸透性は亢進していない、と評価する、前記<11>項に記載の方法。 <7> The indicator substance is methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4, red No. 215, caffeine, ubinal, carotenoid, retinol, and skin moisturizer, skin external preparation, skin cosmetic, or steam The method according to any one of <2> to <6>, wherein the method is at least one selected from the group consisting of an active ingredient or a medicinal ingredient contained in a heating tool.
<8> The method according to any one of <1> to <7>, wherein the moisture content of the skin is measured and analyzed by a skin capacitance measuring unit.
<9> Skin of healthy subject is evaluated, skin moisture after contact with test substance or article is measured by skin capacitance measuring means, and skin moisture after contact with test substance or article is tested. When the amount of change in the moisture content of the skin before contact with a substance or article increases by 8 (AU) or more, preferably 12 (AU) or more, more preferably 16 (AU) or more, the barrier function decreases. Preferably, the method according to <8> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is enhanced.
<10> Skin of healthy subject is evaluated, skin moisture after contact with test substance or article is measured by skin capacitance measuring means, and skin moisture after contact with test substance or article is tested A change of less than 16 (AU) increase, preferably a change of less than 12 (AU) increase, more preferably a change of less than 8 (AU) increase, relative to the amount of moisture in the skin before contact with the substance or article, In the case of <5>, the method according to <8>, wherein the barrier function is not changed, preferably the permeability of the substance from the outside to the inside of the skin is not enhanced.
<11> Up to a depth of 2 to 40 μm from the skin surface, preferably up to a depth of 2 to 35 μm from the skin surface, more preferably up to a depth of 2 to 20 μm from the skin surface, more preferably The water content or water distribution up to a depth of 9 to 15 μm from the skin surface, more preferably a depth of about 13 μm from the skin surface is measured and analyzed by Raman spectroscopy, <1> to < 7. The method according to any one of 7>.
<12> The skin of a healthy person is the subject of evaluation, up to a predetermined depth from the skin surface, preferably up to a depth of 40 μm from the skin surface, more preferably up to a depth of 35 μm from the skin surface, more preferably The skin moisture distribution before and after contact with the test substance or article up to a depth of 20 μm from the skin surface is measured by Raman spectroscopy, and the skin moisture distribution before and after contact with the test substance or article is compared. When the amount of moisture in the skin deeper than the initial stratum corneum thickness is increased by contact with the substance or article, it is evaluated that the barrier function is lowered, preferably the permeability of the substance from the outside to the inside of the skin is increased. The method according to <11> above.
<13> Skin surface moisture after contact with a test substance or article obtained by Raman spectroscopy is 2 with respect to the skin surface moisture before contact with the test substance or article. When it increases to 0.0 times or more, preferably 2.5 times or more, more preferably 3.0 times or more, the barrier function is lowered, preferably the permeability of the substance from the outside of the skin to the inside is increased, The method according to <11>, wherein the method is evaluated.
<14> The skin surface moisture after contact of the test substance or article obtained by Raman spectroscopy is 3 with respect to the skin surface moisture before contact of the test substance or article. If it is less than 0.0 times increase, preferably less than 2.5 times, more preferably less than 2.0 times, the barrier function is not changed, preferably the permeability of the substance from outside the skin to the inside is The method according to <11>, wherein the method is evaluated as not enhanced.
<15> The skin of a healthy person is the subject of evaluation, and the water content of the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the value before contact with the test substance or article. When the water content in the region corresponding to the initial stratum corneum thickness is increased by 8% or more, preferably 11% or more, more preferably 14% or more, the barrier function is lowered, preferably from the outside of the skin to the inside. The method according to <11>, wherein the permeability of the substance is evaluated to be enhanced.
<16> Using healthy skin as an evaluation target, the moisture content of the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the value before contact with the test substance or article. If the amount of water in the region corresponding to the initial stratum corneum thickness is less than 14% increase, preferably less than 11% increase, more preferably less than 8% increase, the barrier function is not changed, Preferably, the method according to <11> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is not enhanced.
<17> The skin of a healthy person is evaluated, and is obtained by Raman spectroscopy up to a predetermined depth from the skin surface, preferably a depth of 2 to 40 μm from the skin surface, more preferably from the skin surface. To a depth of 2 to 35 μm, more to a depth of 2 to 20 μm from the skin surface, more preferably to a depth of 9 to 15 μm from the skin surface, more preferably to a depth of 13 μm from the skin surface. The area under the curve of the moisture content of the skin after contact with the test substance or article up to a certain depth is obtained, and the area under the curve of the skin moisture amount after contact with the test substance or article is the contact of the test substance or article When increased to 1.4 times or more, preferably 1.6 times or more, more preferably 1.8 times or more with respect to the area under the curve of moisture content of the previous skin, the barrier function is lowered, preferably skin From outside to inside The method according to <11> above, wherein it is evaluated that the permeability of the substance to the substance is enhanced.
<18> The skin of a healthy person is evaluated, and obtained by Raman spectroscopy, to a predetermined depth from the skin surface, preferably to a depth of 2 to 40 μm from the skin surface, more preferably from the skin surface. To a depth of 2 to 35 μm, more to a depth of 2 to 20 μm from the skin surface, more preferably to a depth of 9 to 15 μm from the skin surface, more preferably to a depth of 13 μm from the skin surface. The area under the curve of the moisture content of the skin after contact with the test substance or article up to a certain depth is obtained, and the area under the curve of the skin moisture amount after contact with the test substance or article is the contact of the test substance or article If the area under the curve of water content of the previous skin is less than 1.8 times, preferably less than 1.6 times, more preferably less than 1.4 times, the barrier function is not changed, Like Alternatively, the method according to <11> above, wherein it is evaluated that the permeability of the substance from the outside to the inside of the skin is not enhanced.
<19>被験物質又は物品が、ヒトの皮膚のバリア機能に与える影響を評価する方法であって、
被験物質又は物品のヒトの皮膚への接触前と接触後の皮膚の深さ毎の水分量を測定して水分分布を取得し、
接触前と接触後の水分分布における、接触前の初期の角層厚よりも深部の水分量の差を指標として、被験物質又は物品がヒトの皮膚のバリア機能に与える影響を評価する、
被験物質又は物品の評価方法。 <19> A method for evaluating the influence of a test substance or article on the barrier function of human skin,
Obtain water distribution by measuring the amount of water at each skin depth before and after contact of the test substance or article with human skin,
Evaluate the effect of the test substance or article on the barrier function of human skin, using the difference in water content deeper than the initial stratum corneum thickness before contact in the moisture distribution before and after contact,
Evaluation method of test substance or article.
被験物質又は物品のヒトの皮膚への接触前と接触後の皮膚の深さ毎の水分量を測定して水分分布を取得し、
接触前と接触後の水分分布における、接触前の初期の角層厚よりも深部の水分量の差を指標として、被験物質又は物品がヒトの皮膚のバリア機能に与える影響を評価する、
被験物質又は物品の評価方法。 <19> A method for evaluating the influence of a test substance or article on the barrier function of human skin,
Obtain water distribution by measuring the amount of water at each skin depth before and after contact of the test substance or article with human skin,
Evaluate the effect of the test substance or article on the barrier function of human skin, using the difference in water content deeper than the initial stratum corneum thickness before contact in the moisture distribution before and after contact,
Evaluation method of test substance or article.
<20>皮膚表面からの深さ2~40μmの深さまで、好ましくは皮膚表面からの深さ2~35μmの深さまで、より好ましくは皮膚表面からの深さ2~20μmの深さまで、より好ましくは皮膚表面からの深さ9~15μmの深さまで、より好ましくは皮膚表面からの深さ13μm程度の深さまで、の水分量をラマン分光法により測定し解析する、前記<19>項に記載の方法。
<21>健常者の皮膚を評価対象とし、皮膚表面から所定の深さまでにおける、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合、バリア機能が低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<19>又は<20>項に記載の方法。 <20> to a depth of 2 to 40 μm from the skin surface, preferably to a depth of 2 to 35 μm from the skin surface, more preferably to a depth of 2 to 20 μm from the skin surface, more preferably The method according to <19>, wherein the moisture content is measured and analyzed by Raman spectroscopy up to a depth of 9 to 15 μm from the skin surface, more preferably a depth of about 13 μm from the skin surface. .
<21> Skin of a healthy subject is evaluated by measuring the moisture distribution of the skin before and after contact with the test substance or article from the skin surface to a predetermined depth by Raman spectroscopy, and before and after contact with the test substance or article When the moisture content of the skin in the deeper part of the stratum corneum increases than the initial stratum corneum thickness due to contact with the test substance or article, the barrier function decreases, preferably the permeability of the substance from the outside to the inside of the skin The method according to <19> or <20>, wherein the evaluation is as follows.
<21>健常者の皮膚を評価対象とし、皮膚表面から所定の深さまでにおける、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合、バリア機能が低下した、好ましくは皮膚外部から内部への物質の浸透性が亢進した、と評価する、前記<19>又は<20>項に記載の方法。 <20> to a depth of 2 to 40 μm from the skin surface, preferably to a depth of 2 to 35 μm from the skin surface, more preferably to a depth of 2 to 20 μm from the skin surface, more preferably The method according to <19>, wherein the moisture content is measured and analyzed by Raman spectroscopy up to a depth of 9 to 15 μm from the skin surface, more preferably a depth of about 13 μm from the skin surface. .
<21> Skin of a healthy subject is evaluated by measuring the moisture distribution of the skin before and after contact with the test substance or article from the skin surface to a predetermined depth by Raman spectroscopy, and before and after contact with the test substance or article When the moisture content of the skin in the deeper part of the stratum corneum increases than the initial stratum corneum thickness due to contact with the test substance or article, the barrier function decreases, preferably the permeability of the substance from the outside to the inside of the skin The method according to <19> or <20>, wherein the evaluation is as follows.
<22>皮膚水分量又は水分分布を非侵襲的に測定する、前記<1>~<21>のいずれか1項に記載の方法。
<23>被験物質又は物品を負荷していない状態で正常なバリア機能を有するヒト(健常者)の皮膚で評価する、前記<1>~<22>のいずれか1項に記載の方法。
<24>ラマン分光法、皮膚静電容量測定手段、高周波電流測定手段、赤外分光法、及び近赤外分光法からなる群より選ばれる少なくとも1つに従い皮膚の水分量を測定する、前記<1>~<7>、<19>、<22>及び<23>のいずれか1項に記載の方法。
<25>前記被験物質が、水若しくは水蒸気、皮膚化粧料、皮膚保湿剤若しくは皮膚外用剤、蒸気温熱具に含まれる有効成分若しくは薬効成分、生理食塩水、リン酸緩衝生理食塩水、並びに生体由来の分泌物若しくは***物からなる群より選ばれる少なくとも1種である、前記<1>~<24>のいずれか1項に記載の方法。
<26>前記物品が、吸収性物品(好ましくは脱脂綿、生理用ナプキン、紙おむつ、布おむつ、トレーニングパンツ、成人用おむつ、汗止めパット、尿取りパット、産褥パット、パンティーライナー、若しくは吸収パット)、蒸気温熱具、創傷保護材(好ましくは絆創膏、ガーゼ、若しくは包帯)、マスク、又はゴム手袋である、前記<1>~<24>のいずれか1項に記載の方法。
<27>前記吸収性物品が生理食塩水又は擬似尿などを含有する、前記<26>項に記載の方法。
<28>前記被験物質又は物品をヒトの皮膚に接触させる時間が、1時間以上6時間以下、好ましくは2時間以上4時間以下、より好ましくは2.5時間以上3.5時間以下、さらに好ましくは3時間程度、である、前記<1>~<27>のいずれか1項に記載の方法。 <22> The method according to any one of <1> to <21>, wherein the skin moisture content or moisture distribution is noninvasively measured.
<23> The method according to any one of <1> to <22>, wherein the evaluation is performed on the skin of a human (healthy person) having a normal barrier function in a state in which no test substance or article is loaded.
<24> The moisture content of the skin is measured according to at least one selected from the group consisting of Raman spectroscopy, skin capacitance measurement means, high-frequency current measurement means, infrared spectroscopy, and near-infrared spectroscopy, The method according to any one of 1> to <7>, <19>, <22> and <23>.
<25> The test substance is water or water vapor, skin cosmetics, skin moisturizer or external preparation for skin, active ingredient or medicinal ingredient contained in steam heating device, physiological saline, phosphate buffered saline, and biological origin The method according to any one of <1> to <24>, wherein the method is at least one selected from the group consisting of secretions and excrements of
<26> The article is an absorbent article (preferably absorbent cotton, a sanitary napkin, a paper diaper, a cloth diaper, a training pant, an adult diaper, a sweat pad, a urine pad, a postpartum pad, a panty liner, or an absorbent pad), The method according to any one of <1> to <24>, wherein the method is a steam heating device, a wound protection material (preferably a bandage, a gauze, or a bandage), a mask, or a rubber glove.
<27> The method according to <26>, wherein the absorbent article contains physiological saline or simulated urine.
<28> Time for contacting the test substance or article with human skin is 1 hour or more and 6 hours or less, preferably 2 hours or more and 4 hours or less, more preferably 2.5 hours or more and 3.5 hours or less, and further preferably The method according to any one of <1> to <27>, wherein is about 3 hours.
<23>被験物質又は物品を負荷していない状態で正常なバリア機能を有するヒト(健常者)の皮膚で評価する、前記<1>~<22>のいずれか1項に記載の方法。
<24>ラマン分光法、皮膚静電容量測定手段、高周波電流測定手段、赤外分光法、及び近赤外分光法からなる群より選ばれる少なくとも1つに従い皮膚の水分量を測定する、前記<1>~<7>、<19>、<22>及び<23>のいずれか1項に記載の方法。
<25>前記被験物質が、水若しくは水蒸気、皮膚化粧料、皮膚保湿剤若しくは皮膚外用剤、蒸気温熱具に含まれる有効成分若しくは薬効成分、生理食塩水、リン酸緩衝生理食塩水、並びに生体由来の分泌物若しくは***物からなる群より選ばれる少なくとも1種である、前記<1>~<24>のいずれか1項に記載の方法。
<26>前記物品が、吸収性物品(好ましくは脱脂綿、生理用ナプキン、紙おむつ、布おむつ、トレーニングパンツ、成人用おむつ、汗止めパット、尿取りパット、産褥パット、パンティーライナー、若しくは吸収パット)、蒸気温熱具、創傷保護材(好ましくは絆創膏、ガーゼ、若しくは包帯)、マスク、又はゴム手袋である、前記<1>~<24>のいずれか1項に記載の方法。
<27>前記吸収性物品が生理食塩水又は擬似尿などを含有する、前記<26>項に記載の方法。
<28>前記被験物質又は物品をヒトの皮膚に接触させる時間が、1時間以上6時間以下、好ましくは2時間以上4時間以下、より好ましくは2.5時間以上3.5時間以下、さらに好ましくは3時間程度、である、前記<1>~<27>のいずれか1項に記載の方法。 <22> The method according to any one of <1> to <21>, wherein the skin moisture content or moisture distribution is noninvasively measured.
<23> The method according to any one of <1> to <22>, wherein the evaluation is performed on the skin of a human (healthy person) having a normal barrier function in a state in which no test substance or article is loaded.
<24> The moisture content of the skin is measured according to at least one selected from the group consisting of Raman spectroscopy, skin capacitance measurement means, high-frequency current measurement means, infrared spectroscopy, and near-infrared spectroscopy, The method according to any one of 1> to <7>, <19>, <22> and <23>.
<25> The test substance is water or water vapor, skin cosmetics, skin moisturizer or external preparation for skin, active ingredient or medicinal ingredient contained in steam heating device, physiological saline, phosphate buffered saline, and biological origin The method according to any one of <1> to <24>, wherein the method is at least one selected from the group consisting of secretions and excrements of
<26> The article is an absorbent article (preferably absorbent cotton, a sanitary napkin, a paper diaper, a cloth diaper, a training pant, an adult diaper, a sweat pad, a urine pad, a postpartum pad, a panty liner, or an absorbent pad), The method according to any one of <1> to <24>, wherein the method is a steam heating device, a wound protection material (preferably a bandage, a gauze, or a bandage), a mask, or a rubber glove.
<27> The method according to <26>, wherein the absorbent article contains physiological saline or simulated urine.
<28> Time for contacting the test substance or article with human skin is 1 hour or more and 6 hours or less, preferably 2 hours or more and 4 hours or less, more preferably 2.5 hours or more and 3.5 hours or less, and further preferably The method according to any one of <1> to <27>, wherein is about 3 hours.
<29>被験物質又は物品のヒトの皮膚への接触による皮膚の水分量又は水分分布の変化量と皮膚のバリア機能との関連性に関する情報を格納し、関連性に関する情報に基づき、被験物質又は物品の接触後の皮膚の水分量又は水分分布から、被験物質又は物品が、皮膚のバリア機能に与える影響を評価するためのシステム。
<29> Stores information on the relationship between the amount of change in the moisture content or distribution of skin due to contact of the test substance or article with human skin, and the barrier function of the skin. A system for evaluating the influence of a test substance or an article on the barrier function of the skin from the moisture content or moisture distribution of the skin after contact with the article.
以下、本発明を実施例に基づきさらに詳細に説明するが、本発明はこれに限定されるものではない。
Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited thereto.
試験例1 水分負荷前後のニコチン酸メチルの透過性と皮膚の水分量の変化の測定
(1)紅斑スコアの測定
健常な男性、女性10名(20代~40代)を被験者とした。前腕内側に異なる量の水分を負荷し、水分負荷後のニコチン酸メチルの透過性を評価した。測定は、24~25℃、相対湿度40~50%の環境可変室で、20分順化した後に行った。 Test Example 1 Measurement of change in permeability of methyl nicotinate and moisture content of skin before and after water load (1) Measurement of erythema score Healthy males and 10 females (20s to 40s) were subjects. A different amount of water was loaded on the inner side of the forearm, and the permeability of methyl nicotinate after the water load was evaluated. The measurement was carried out after acclimatization for 20 minutes in an environment variable room at 24 to 25 ° C. and a relative humidity of 40 to 50%.
(1)紅斑スコアの測定
健常な男性、女性10名(20代~40代)を被験者とした。前腕内側に異なる量の水分を負荷し、水分負荷後のニコチン酸メチルの透過性を評価した。測定は、24~25℃、相対湿度40~50%の環境可変室で、20分順化した後に行った。 Test Example 1 Measurement of change in permeability of methyl nicotinate and moisture content of skin before and after water load (1) Measurement of erythema score Healthy males and 10 females (20s to 40s) were subjects. A different amount of water was loaded on the inner side of the forearm, and the permeability of methyl nicotinate after the water load was evaluated. The measurement was carried out after acclimatization for 20 minutes in an environment variable room at 24 to 25 ° C. and a relative humidity of 40 to 50%.
2.5cm×2.5cmのメンバンカット綿(白十字社製)に、脱脂綿1cm2当たりの水分負荷量が0mL(0mL/cm2)、0.02mL(0.02mL/cm2)、0.04mL(0.04mL/cm2)、0.20mL(0.20mL/cm2)となるように蒸留水を含ませ、これを前腕内側にのせ透湿性絆創膏(商品名:スキナゲート)で周囲を固定し3時間貼付した。メンバンカット綿から水分が漏れるのを防ぐため、メンバンカット綿の皮膚と接しない面はドレッシング材(商品名:テガダームTM トランスペアレント ドレッシング)で覆い、貼付中に皮膚が蒸れるのを防ぐためドレッシング材は注射針29G(TERUMO, Japan)で1cm2当たり8個、穴をあけた。メンバンカット綿を貼付している間、試験参加者には運動や屋外での長時間の作業は禁止した。
Moisture load per cm 2 of absorbent cotton is 0 mL (0 mL / cm 2 ), 0.02 mL (0.02 mL / cm 2 ), 0 on 2.5 cm × 2.5 cm member cut cotton (manufactured by White Cross) Add distilled water to make 0.04 mL (0.04 mL / cm 2 ), 0.20 mL (0.20 mL / cm 2 ), place this on the inner side of the forearm, and use a moisture-permeable adhesive bandage (trade name: Skinnagate) to surround it. Fixed and affixed for 3 hours. In order to prevent moisture from leaking from the member cut cotton, the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm ™ Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
メンバンカット綿を除去した直後に、0.007質量%ニコチン酸メチル水溶液5.0μLを含ませた、直径5mmにくり抜いたろ紙(商品名:Advantec No2)を試験部位に15秒間負荷した。ニコチン酸メチルの負荷から30分後に、生じた紅斑を下記の評価基準に従い目視判定を行った。
目視判定により得られた紅斑スコアから、各水分負荷量でのニコチン酸メチルの透過性を評価した。統計解析にはIBM SPSSバージョン23のソフトウェアを使用した。
その結果を表1に示す。
(評価基準)
スコア0 :反応なし。
スコア0.5:若干紅い。
スコア1 :ろ紙の面積の半分以上紅い。
スコア2 :ろ紙を当てた面積がはっきり紅い。
スコア3 :ろ紙の面積より大きく紅い。
スコア4 :紅みがろ紙の面積より大きく、腫れている。 Immediately after removing the member cut cotton, a filter paper (trade name: Advantec No. 2) hollowed out to a diameter of 5 mm containing 5.0 μL of 0.007 mass% methyl nicotinate aqueous solution was loaded on the test site for 15 seconds. After 30 minutes from the load of methyl nicotinate, the resulting erythema was visually determined according to the following evaluation criteria.
From the erythema score obtained by visual determination, the permeability of methyl nicotinate at each water load was evaluated. IBM SPSS version 23 software was used for statistical analysis.
The results are shown in Table 1.
(Evaluation criteria)
Score 0: No response.
Score 0.5: Slightly red.
Score 1: More than half of the area of the filter paper is red.
Score 2: The area on which the filter paper is applied is clearly red.
Score 3: larger than the area of the filter paper.
Score 4: Red area is larger than the area of the filter paper and swollen.
目視判定により得られた紅斑スコアから、各水分負荷量でのニコチン酸メチルの透過性を評価した。統計解析にはIBM SPSSバージョン23のソフトウェアを使用した。
その結果を表1に示す。
(評価基準)
スコア0 :反応なし。
スコア0.5:若干紅い。
スコア1 :ろ紙の面積の半分以上紅い。
スコア2 :ろ紙を当てた面積がはっきり紅い。
スコア3 :ろ紙の面積より大きく紅い。
スコア4 :紅みがろ紙の面積より大きく、腫れている。 Immediately after removing the member cut cotton, a filter paper (trade name: Advantec No. 2) hollowed out to a diameter of 5 mm containing 5.0 μL of 0.007 mass% methyl nicotinate aqueous solution was loaded on the test site for 15 seconds. After 30 minutes from the load of methyl nicotinate, the resulting erythema was visually determined according to the following evaluation criteria.
From the erythema score obtained by visual determination, the permeability of methyl nicotinate at each water load was evaluated. IBM SPSS version 23 software was used for statistical analysis.
The results are shown in Table 1.
(Evaluation criteria)
Score 0: No response.
Score 0.5: Slightly red.
Score 1: More than half of the area of the filter paper is red.
Score 2: The area on which the filter paper is applied is clearly red.
Score 3: larger than the area of the filter paper.
Score 4: Red area is larger than the area of the filter paper and swollen.
表1に示すように、水分の負荷量が0.00mL/cm2又は0.02mL/cm2の場合と比較して、0.04mL/cm2又は0.20mL/cm2の水分を負荷することで、ニコチン酸メチルによる紅斑スコアが有意に上昇した。すなわち、一定量以上の水分を皮膚に負荷することで、ニコチン酸メチルの皮膚への透過性が有意に亢進した。
本試験例の健常なヒトの前腕において、水分の負荷量が少なくとも0.04mL/cm2の場合に皮膚バリア機能が低下していると判断することができる。よって、水分負荷量が0.04mL/cm2の場合の皮膚の状態を、皮膚のバリア機能を評価するための基準状態とすることができる。 As shown in Table 1, the load amount of water as compared with the case of 0.00mL / cm 2 or 0.02 mL / cm 2, to load a moisture 0.04 mL / cm 2 or 0.20 mL / cm 2 As a result, the erythema score due to methyl nicotinate was significantly increased. That is, the permeation of methyl nicotinate into the skin was significantly enhanced by applying a certain amount or more of moisture to the skin.
In the healthy human forearm of this test example, it can be determined that the skin barrier function is lowered when the water load is at least 0.04 mL / cm 2 . Therefore, the state of the skin when the moisture load is 0.04 mL / cm 2 can be set as a reference state for evaluating the barrier function of the skin.
本試験例の健常なヒトの前腕において、水分の負荷量が少なくとも0.04mL/cm2の場合に皮膚バリア機能が低下していると判断することができる。よって、水分負荷量が0.04mL/cm2の場合の皮膚の状態を、皮膚のバリア機能を評価するための基準状態とすることができる。 As shown in Table 1, the load amount of water as compared with the case of 0.00mL / cm 2 or 0.02 mL / cm 2, to load a moisture 0.04 mL / cm 2 or 0.20 mL / cm 2 As a result, the erythema score due to methyl nicotinate was significantly increased. That is, the permeation of methyl nicotinate into the skin was significantly enhanced by applying a certain amount or more of moisture to the skin.
In the healthy human forearm of this test example, it can be determined that the skin barrier function is lowered when the water load is at least 0.04 mL / cm 2 . Therefore, the state of the skin when the moisture load is 0.04 mL / cm 2 can be set as a reference state for evaluating the barrier function of the skin.
負荷されたニコチン酸メチルが皮膚へ浸透又は通過し、末梢血管に働きかけ血管拡張作用を示すと、皮膚上で紅斑が観察される。表1に示すように、水分負荷前の皮膚(水分負荷量:0mL/cm2)では、ニコチン酸メチルを負荷しても紅斑は観察されにくい。これは、ニコチン酸メチルが皮膚内へ浸透されにくく、末梢血管まで到達できないためと考えられる。すなわち、水分負荷前の皮膚では、バリア機能が維持されていると推察できる。一方、一定量以上の水分負荷後の皮膚では、ニコチン酸メチルの負荷によって紅斑が観察される。この結果から、水分を負荷することでニコチン酸メチルが皮膚へ浸透又は通過した、すなわち皮膚のバリア機能が低下したと判断できる。
Erythema is observed on the skin when the loaded methyl nicotinate penetrates or passes through the skin and acts on the peripheral blood vessels to exert a vasodilating action. As shown in Table 1, erythema is hardly observed on the skin before water load (water load: 0 mL / cm 2 ) even when methyl nicotinate is loaded. This is probably because methyl nicotinate is difficult to penetrate into the skin and cannot reach the peripheral blood vessels. That is, it can be inferred that the barrier function is maintained in the skin before moisture loading. On the other hand, erythema is observed in the skin after a certain amount of water load due to the load of methyl nicotinate. From this result, it can be determined that loading with moisture caused methyl nicotinate to penetrate or pass through the skin, that is, the barrier function of the skin was reduced.
(2)皮膚静電容量測定装置を用いた皮膚水分量の測定
2.5cm×2.5cmのメンバンカット綿(白十字社製)に、脱脂綿1cm2当たりの水分負荷量が0mL(0mL/cm2)、0.01mL(0.01mL/cm2)、0.02mL(0.02mL/cm2)、0.03mL(0.03mL/cm2)、0.04mL(0.04mL/cm2)、0.06mL(0.06mL/cm2)、0.10mL(0.10mL/cm2)、0.20mL(0.20mL/cm2)となるように蒸留水を含ませ、これを前記被験者の前腕内側にのせ透湿性絆創膏(商品名:スキナゲート)で周囲を固定し3時間貼付した。メンバンカット綿から水分が漏れるのを防ぐため、メンバンカット綿の皮膚と接しない面はドレッシング材(商品名:テガダームTM トランスペアレント ドレッシング)で覆い、貼付中に皮膚が蒸れるのを防ぐためドレッシング材は注射針29G(TERUMO, Japan)で1cm2当たり8個、穴をあけた。メンバンカット綿を貼付している間、試験参加者には運動や屋外での長時間の作業は禁止した。 (2) Measurement of skin moisture using a skin capacitance measuring device A moisture load per cm 2 of absorbent cotton is 0 mL (0 mL / 0) on a 2.5 cm × 2.5 cm member cut cotton (manufactured by White Cross). cm 2), 0.01mL (0.01mL / cm 2), 0.02mL (0.02mL / cm 2), 0.03mL (0.03mL / cm 2), 0.04mL (0.04mL / cm 2 ), 0.06 mL (0.06 mL / cm 2 ), 0.10 mL (0.10 mL / cm 2 ), and 0.20 mL (0.20 mL / cm 2 ) in distilled water, The circumference was fixed with a moisture-permeable adhesive plaster (trade name: Skinnagate) on the inner side of the subject's forearm and applied for 3 hours. In order to prevent moisture from leaking from the member cut cotton, the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm ™ Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
2.5cm×2.5cmのメンバンカット綿(白十字社製)に、脱脂綿1cm2当たりの水分負荷量が0mL(0mL/cm2)、0.01mL(0.01mL/cm2)、0.02mL(0.02mL/cm2)、0.03mL(0.03mL/cm2)、0.04mL(0.04mL/cm2)、0.06mL(0.06mL/cm2)、0.10mL(0.10mL/cm2)、0.20mL(0.20mL/cm2)となるように蒸留水を含ませ、これを前記被験者の前腕内側にのせ透湿性絆創膏(商品名:スキナゲート)で周囲を固定し3時間貼付した。メンバンカット綿から水分が漏れるのを防ぐため、メンバンカット綿の皮膚と接しない面はドレッシング材(商品名:テガダームTM トランスペアレント ドレッシング)で覆い、貼付中に皮膚が蒸れるのを防ぐためドレッシング材は注射針29G(TERUMO, Japan)で1cm2当たり8個、穴をあけた。メンバンカット綿を貼付している間、試験参加者には運動や屋外での長時間の作業は禁止した。 (2) Measurement of skin moisture using a skin capacitance measuring device A moisture load per cm 2 of absorbent cotton is 0 mL (0 mL / 0) on a 2.5 cm × 2.5 cm member cut cotton (manufactured by White Cross). cm 2), 0.01mL (0.01mL / cm 2), 0.02mL (0.02mL / cm 2), 0.03mL (0.03mL / cm 2), 0.04mL (0.04mL / cm 2 ), 0.06 mL (0.06 mL / cm 2 ), 0.10 mL (0.10 mL / cm 2 ), and 0.20 mL (0.20 mL / cm 2 ) in distilled water, The circumference was fixed with a moisture-permeable adhesive plaster (trade name: Skinnagate) on the inner side of the subject's forearm and applied for 3 hours. In order to prevent moisture from leaking from the member cut cotton, the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm ™ Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
メンバンカット綿の貼付前と、メンバンカット綿の除去直後に、メンバンカット綿を貼付した部位の皮膚の水分量を皮膚静電容量測定装置(商品名:コルネオメーター CM-825、Courage+Khazaka Germany社製)で測定した。そして、メンバンカット綿の除去直後の値から、メンバンカット綿の貼付前の値を差し引いた値を算出し、角層水分量の変化量を測定した。統計解析にはIBM SPSSバージョン23のソフトウェアを使用した。
その結果を表2に示す。 Skin capacitance measuring device (trade name: Corneometer CM-825, Courage +) before and after removing the member cut cotton, and measuring the moisture content of the skin where the member cut cotton is applied. Measured by Khazaka Germany). Then, a value obtained by subtracting the value before application of the member cut cotton from the value immediately after the removal of the member cut cotton was calculated, and the amount of change in the stratum corneum moisture content was measured. IBM SPSS version 23 software was used for statistical analysis.
The results are shown in Table 2.
その結果を表2に示す。 Skin capacitance measuring device (trade name: Corneometer CM-825, Courage +) before and after removing the member cut cotton, and measuring the moisture content of the skin where the member cut cotton is applied. Measured by Khazaka Germany). Then, a value obtained by subtracting the value before application of the member cut cotton from the value immediately after the removal of the member cut cotton was calculated, and the amount of change in the stratum corneum moisture content was measured. IBM SPSS version 23 software was used for statistical analysis.
The results are shown in Table 2.
表2に示すように、水分の負荷量が0mL/cm2から0.06mL/cm2までの間は、皮膚の水分量は水分の負荷量に依存的に増加した。特に、水分の負荷量が0.03mL/cm2以上の場合、皮膚の水分量は有意に増加した。一方、水分の負荷量が0.06mL/cm2以上の場合は、皮膚の水分量はほぼ一定の数値となった。
以上の結果から、水分の負荷量が一定量に達するまでは、皮膚の水分量は水分の負荷量に依存して増加する。特に、水分の負荷量が一定値以上増えると皮膚の水分量は有意に増加する。
さらに前述のように、水分の負荷量が少なくとも0.04mL/cm2の場合に皮膚バリア機能が低下していると判断でき、水分負荷量が0.04mL/cm2の場合の皮膚の状態を皮膚のバリア機能を評価するための基準状態とした。よって、この基準状態での皮膚水分量の変化量を、本発明の評価方法における基準値とすることができる。 As shown in Table 2, when the water load was from 0 mL / cm 2 to 0.06 mL / cm 2 , the skin water content increased depending on the water load. In particular, when the moisture load was 0.03 mL / cm 2 or more, the moisture content of the skin was significantly increased. On the other hand, when the moisture load was 0.06 mL / cm 2 or more, the moisture content of the skin was almost constant.
From the above results, the moisture content of the skin increases depending on the moisture loading amount until the moisture loading amount reaches a certain amount. In particular, when the moisture load increases by a certain value or more, the moisture content of the skin increases significantly.
Further, as described above, when the load amount of water of at least 0.04 mL / cm 2 can be determined that the skin barrier function is impaired, the state of the skin when the water loading amount of 0.04 mL / cm 2 A reference state for evaluating the skin barrier function was used. Therefore, the amount of change in the amount of skin moisture in this reference state can be used as the reference value in the evaluation method of the present invention.
以上の結果から、水分の負荷量が一定量に達するまでは、皮膚の水分量は水分の負荷量に依存して増加する。特に、水分の負荷量が一定値以上増えると皮膚の水分量は有意に増加する。
さらに前述のように、水分の負荷量が少なくとも0.04mL/cm2の場合に皮膚バリア機能が低下していると判断でき、水分負荷量が0.04mL/cm2の場合の皮膚の状態を皮膚のバリア機能を評価するための基準状態とした。よって、この基準状態での皮膚水分量の変化量を、本発明の評価方法における基準値とすることができる。 As shown in Table 2, when the water load was from 0 mL / cm 2 to 0.06 mL / cm 2 , the skin water content increased depending on the water load. In particular, when the moisture load was 0.03 mL / cm 2 or more, the moisture content of the skin was significantly increased. On the other hand, when the moisture load was 0.06 mL / cm 2 or more, the moisture content of the skin was almost constant.
From the above results, the moisture content of the skin increases depending on the moisture loading amount until the moisture loading amount reaches a certain amount. In particular, when the moisture load increases by a certain value or more, the moisture content of the skin increases significantly.
Further, as described above, when the load amount of water of at least 0.04 mL / cm 2 can be determined that the skin barrier function is impaired, the state of the skin when the water loading amount of 0.04 mL / cm 2 A reference state for evaluating the skin barrier function was used. Therefore, the amount of change in the amount of skin moisture in this reference state can be used as the reference value in the evaluation method of the present invention.
(3)共焦点ラマン分光顕微鏡を用いた皮膚水分量の測定
2.5cm×2.5cmのメンバンカット綿(白十字社製)に、脱脂綿1cm2当たりの水分負荷量が0mL(0mL/cm2)、0.02mL(0.02mL/cm2)、0.04mL(0.04mL/cm2)、0.20mL(0.20mL/cm2)となるように蒸留水を含ませ、これを前記被験者の前腕内側にのせ透湿性絆創膏(商品名:スキナゲート)で周囲を固定し3時間貼付した。メンバンカット綿から水分が漏れるのを防ぐため、メンバンカット綿の皮膚と接しない面はドレッシング材(商品名:テガダームTM トランスペアレント ドレッシング)で覆い、貼付中に皮膚が蒸れるのを防ぐためドレッシング材は注射針29G(TERUMO, Japan)で1cm2当たり8個、穴をあけた。メンバンカット綿を貼付している間、試験参加者には運動や屋外での長時間の作業は禁止した。 (3) Measurement of skin water content using confocal Raman spectroscopic microscope A water load per 1 cm 2 of absorbent cotton is 0 mL (0 mL / cm) on 2.5 cm × 2.5 cm member cut cotton (manufactured by White Cross). 2 ) Distilled water was added to make 0.02 mL (0.02 mL / cm 2 ), 0.04 mL (0.04 mL / cm 2 ), and 0.20 mL (0.20 mL / cm 2 ). The periphery was fixed on the inner side of the subject's forearm with a moisture-permeable adhesive bandage (trade name: Skinnagate) and applied for 3 hours. In order to prevent moisture from leaking from the member cut cotton, the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm ™ Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
2.5cm×2.5cmのメンバンカット綿(白十字社製)に、脱脂綿1cm2当たりの水分負荷量が0mL(0mL/cm2)、0.02mL(0.02mL/cm2)、0.04mL(0.04mL/cm2)、0.20mL(0.20mL/cm2)となるように蒸留水を含ませ、これを前記被験者の前腕内側にのせ透湿性絆創膏(商品名:スキナゲート)で周囲を固定し3時間貼付した。メンバンカット綿から水分が漏れるのを防ぐため、メンバンカット綿の皮膚と接しない面はドレッシング材(商品名:テガダームTM トランスペアレント ドレッシング)で覆い、貼付中に皮膚が蒸れるのを防ぐためドレッシング材は注射針29G(TERUMO, Japan)で1cm2当たり8個、穴をあけた。メンバンカット綿を貼付している間、試験参加者には運動や屋外での長時間の作業は禁止した。 (3) Measurement of skin water content using confocal Raman spectroscopic microscope A water load per 1 cm 2 of absorbent cotton is 0 mL (0 mL / cm) on 2.5 cm × 2.5 cm member cut cotton (manufactured by White Cross). 2 ) Distilled water was added to make 0.02 mL (0.02 mL / cm 2 ), 0.04 mL (0.04 mL / cm 2 ), and 0.20 mL (0.20 mL / cm 2 ). The periphery was fixed on the inner side of the subject's forearm with a moisture-permeable adhesive bandage (trade name: Skinnagate) and applied for 3 hours. In order to prevent moisture from leaking from the member cut cotton, the surface of the member cut cotton that does not come into contact with the skin is covered with a dressing material (trade name: Tegaderm ™ Transparent Dressing), and the dressing material is used to prevent the skin from stuffy during application. Made 8 holes per 1 cm 2 with a needle 29G (TERUMO, Japan). While applying the member cut cotton, the participants were prohibited from exercising or working outdoors for a long time.
メンバンカット綿の貼付前と、メンバンカット綿の除去直後に、メンバンカット綿を貼付した部位の皮膚の水分量を共焦点ラマン分光顕微鏡(商品名:ナノファインダー30、東京インスツルメンツ社製)で測定した。なお測定条件は以下の通りである。
(測定条件)
励起波長:633nm
積算時間:3秒/1スペクトル
回析格子:150本/mm
対物レンズ:100倍、油浸、NA=1.3(ニコン)
深さ方向スキャン条件:2μm間隔で計21点
1部位あたり5回測定 Confocal Raman spectroscopic microscope (trade name:Nanofinder 30, manufactured by Tokyo Instruments Co., Ltd.) was used to examine the moisture content of the skin where the member cut cotton was applied before and after removing the member cut cotton. Measured with Measurement conditions are as follows.
(Measurement condition)
Excitation wavelength: 633 nm
Integration time: 3 seconds / 1 spectral diffraction grating: 150 lines / mm
Objective lens: 100 times, oil immersion, NA = 1.3 (Nikon)
Depth scanning condition: 5 measurements per 2 sites at 2μm intervals
(測定条件)
励起波長:633nm
積算時間:3秒/1スペクトル
回析格子:150本/mm
対物レンズ:100倍、油浸、NA=1.3(ニコン)
深さ方向スキャン条件:2μm間隔で計21点
1部位あたり5回測定 Confocal Raman spectroscopic microscope (trade name:
(Measurement condition)
Excitation wavelength: 633 nm
Integration time: 3 seconds / 1 spectral diffraction grating: 150 lines / mm
Objective lens: 100 times, oil immersion, NA = 1.3 (Nikon)
Depth scanning condition: 5 measurements per 2 sites at 2μm intervals
共焦点ラマン分光顕微鏡を用いた測定で得られたラマンスペクトルから水分量を近似的に算出する方法は、特開2012-50739号公報に記載の方法を参照して行った。
さらに、IFSCC Magazine, 2009, vol. 12(1), p. 9-15に記載の方法に準じて、ラマン分光法により取得した水分プロファイルを2本の1次直線で近似し、その交点までの深さを初期の角層厚(メンバンカット綿の貼付前の角層厚)として定義した。 A method of approximately calculating the amount of water from the Raman spectrum obtained by measurement using a confocal Raman spectroscopic microscope was performed by referring to the method described in JP2012-50739A.
Furthermore, according to the method described in IFSCC Magazine, 2009, vol. 12 (1), p. 9-15, the moisture profile obtained by Raman spectroscopy is approximated by two linear lines, The depth was defined as the initial stratum corneum thickness (the stratum corneum thickness prior to the application of the member cut cotton).
さらに、IFSCC Magazine, 2009, vol. 12(1), p. 9-15に記載の方法に準じて、ラマン分光法により取得した水分プロファイルを2本の1次直線で近似し、その交点までの深さを初期の角層厚(メンバンカット綿の貼付前の角層厚)として定義した。 A method of approximately calculating the amount of water from the Raman spectrum obtained by measurement using a confocal Raman spectroscopic microscope was performed by referring to the method described in JP2012-50739A.
Furthermore, according to the method described in IFSCC Magazine, 2009, vol. 12 (1), p. 9-15, the moisture profile obtained by Raman spectroscopy is approximated by two linear lines, The depth was defined as the initial stratum corneum thickness (the stratum corneum thickness prior to the application of the member cut cotton).
次に、深さ方向の皮膚内の水分量分布をグラフにプロットした。その結果を図2(a)~(d)に示す。図2(a)~(d)に示すグラフにおいて、X軸は皮膚の深さ(μm)、Y軸は水分量(%)を示す。
図2(a)に示すグラフより求められた水分負荷前の平均の角層厚は13μmであった。そして、図2(b)に示すように、水分負荷量が0.02mL/cm2の場合、水分負荷前と比較して、水分負荷前の角層厚に相当する13μmまでの領域で、皮膚の水分量の増加を確認したが、水負荷前の角層厚よりも深いところの水分量の増加は確認されなかった。
一方で、紅斑スコアに関する前述の実験結果において皮膚のバリア機能が低下し、物質の浸透性の亢進が観察された、水負荷量が0.04mL/cm2以上の場合、水分負荷前の初期の角層厚よりも深いところの皮膚の水分量が増加した(図2(c)及び(d)参照)。水分負荷前の角層厚よりも深いところの水分量が増加することは、角層が膨潤したためと推察でき、より物質が浸透しやすくなったと評価できる。
よって、皮膚のバリア機能に与える影響を評価するための指標として、水分分布を用いて初期の角層厚よりも深部の水分量の差を用いることができる。 Next, the moisture content distribution in the skin in the depth direction was plotted on a graph. The results are shown in FIGS. 2 (a) to (d). In the graphs shown in FIGS. 2A to 2D, the X-axis indicates the skin depth (μm), and the Y-axis indicates the water content (%).
The average thickness of the stratum corneum before the water load determined from the graph shown in FIG. 2A was 13 μm. As shown in FIG. 2 (b), when the moisture load is 0.02 mL / cm 2 , the skin has an area up to 13 μm corresponding to the stratum corneum thickness before moisture load compared to before moisture load. An increase in the amount of water was confirmed, but an increase in the amount of water deeper than the stratum corneum thickness before water loading was not confirmed.
On the other hand, when the skin barrier function is reduced in the above-described experimental results regarding the erythema score and an increase in the permeability of the substance is observed, and the water load is 0.04 mL / cm 2 or more, the initial state before the water load The amount of water in the skin deeper than the stratum corneum thickness increased (see FIGS. 2C and 2D). It can be inferred that the amount of water deeper than the stratum corneum thickness before moisture loading is due to swelling of the stratum corneum, and it can be evaluated that the substance is more easily penetrated.
Therefore, as an index for evaluating the influence on the barrier function of the skin, the difference in the moisture content in the deeper part than the initial stratum corneum thickness can be used using the moisture distribution.
図2(a)に示すグラフより求められた水分負荷前の平均の角層厚は13μmであった。そして、図2(b)に示すように、水分負荷量が0.02mL/cm2の場合、水分負荷前と比較して、水分負荷前の角層厚に相当する13μmまでの領域で、皮膚の水分量の増加を確認したが、水負荷前の角層厚よりも深いところの水分量の増加は確認されなかった。
一方で、紅斑スコアに関する前述の実験結果において皮膚のバリア機能が低下し、物質の浸透性の亢進が観察された、水負荷量が0.04mL/cm2以上の場合、水分負荷前の初期の角層厚よりも深いところの皮膚の水分量が増加した(図2(c)及び(d)参照)。水分負荷前の角層厚よりも深いところの水分量が増加することは、角層が膨潤したためと推察でき、より物質が浸透しやすくなったと評価できる。
よって、皮膚のバリア機能に与える影響を評価するための指標として、水分分布を用いて初期の角層厚よりも深部の水分量の差を用いることができる。 Next, the moisture content distribution in the skin in the depth direction was plotted on a graph. The results are shown in FIGS. 2 (a) to (d). In the graphs shown in FIGS. 2A to 2D, the X-axis indicates the skin depth (μm), and the Y-axis indicates the water content (%).
The average thickness of the stratum corneum before the water load determined from the graph shown in FIG. 2A was 13 μm. As shown in FIG. 2 (b), when the moisture load is 0.02 mL / cm 2 , the skin has an area up to 13 μm corresponding to the stratum corneum thickness before moisture load compared to before moisture load. An increase in the amount of water was confirmed, but an increase in the amount of water deeper than the stratum corneum thickness before water loading was not confirmed.
On the other hand, when the skin barrier function is reduced in the above-described experimental results regarding the erythema score and an increase in the permeability of the substance is observed, and the water load is 0.04 mL / cm 2 or more, the initial state before the water load The amount of water in the skin deeper than the stratum corneum thickness increased (see FIGS. 2C and 2D). It can be inferred that the amount of water deeper than the stratum corneum thickness before moisture loading is due to swelling of the stratum corneum, and it can be evaluated that the substance is more easily penetrated.
Therefore, as an index for evaluating the influence on the barrier function of the skin, the difference in the moisture content in the deeper part than the initial stratum corneum thickness can be used using the moisture distribution.
表面水分量(深さ2μmにおける水分量)の変化量(Δ表面水分量)を算出した。
なお、対物レンズの窓材と皮膚の境界(皮膚のはじまり)を区別して皮膚の表面位置を決定するため、皮膚に含まれるタンパク質のCH伸縮振動由来の信号を目安にした。以下、その詳細を説明する。
ラマンスペクトルのタンパク質のCH伸縮振動由来の信号強度は、前述のように、対物レンズの窓材と皮膚の境界よりやや皮膚側で最大値を示した後に、深部にいくほど信号強度が低下する。窓材にはタンパク質のCH伸縮振動由来の信号は存在しないため、レーザーの焦点が窓材の内にあるときはCH伸縮振動由来の信号強度は0になる。一方、皮膚内にレーザーの焦点があるときのCH伸縮振動由来の信号強度を100とすると、窓材と皮膚の界面に焦点がきた場合、CH伸縮振動由来の信号強度は50となる。ただし、実際には窓材にはCH伸縮振動由来の信号強度が弱いながらも検出されることがある。よって、本発明で測定に用いた共焦点ラマン分光装置であるナノファインダーの空間分解能を考慮し、実際に測定した値である表面から深さ2μmの部位の水分量を「表面水分量」とした。
その結果を表3に示す。 The amount of change (Δ surface water content) of the surface water content (water content at a depth of 2 μm) was calculated.
In order to determine the surface position of the skin by distinguishing the boundary between the window material of the objective lens and the skin (the beginning of the skin), a signal derived from CH stretching vibration of protein contained in the skin was used as a guide. Details will be described below.
As described above, the signal intensity derived from the CH stretching vibration of the protein of the Raman spectrum shows a maximum value on the skin side slightly from the boundary between the window material of the objective lens and the skin, and then the signal intensity decreases as it goes deeper. Since there is no signal derived from protein CH stretching vibration in the window material, the signal intensity derived from CH stretching vibration is 0 when the focal point of the laser is within the window material. On the other hand, when the signal intensity derived from the CH stretching vibration when the laser is focused in the skin is 100, the signal intensity derived from the CH stretching vibration is 50 when the focus comes to the interface between the window material and the skin. In practice, however, the window material may be detected even though the signal intensity derived from the CH stretching vibration is weak. Therefore, in consideration of the spatial resolution of the nano finder, which is the confocal Raman spectroscopic apparatus used for the measurement in the present invention, the water content at a portion 2 μm deep from the surface, which is the actual measurement value, is defined as “surface water content”. .
The results are shown in Table 3.
なお、対物レンズの窓材と皮膚の境界(皮膚のはじまり)を区別して皮膚の表面位置を決定するため、皮膚に含まれるタンパク質のCH伸縮振動由来の信号を目安にした。以下、その詳細を説明する。
ラマンスペクトルのタンパク質のCH伸縮振動由来の信号強度は、前述のように、対物レンズの窓材と皮膚の境界よりやや皮膚側で最大値を示した後に、深部にいくほど信号強度が低下する。窓材にはタンパク質のCH伸縮振動由来の信号は存在しないため、レーザーの焦点が窓材の内にあるときはCH伸縮振動由来の信号強度は0になる。一方、皮膚内にレーザーの焦点があるときのCH伸縮振動由来の信号強度を100とすると、窓材と皮膚の界面に焦点がきた場合、CH伸縮振動由来の信号強度は50となる。ただし、実際には窓材にはCH伸縮振動由来の信号強度が弱いながらも検出されることがある。よって、本発明で測定に用いた共焦点ラマン分光装置であるナノファインダーの空間分解能を考慮し、実際に測定した値である表面から深さ2μmの部位の水分量を「表面水分量」とした。
その結果を表3に示す。 The amount of change (Δ surface water content) of the surface water content (water content at a depth of 2 μm) was calculated.
In order to determine the surface position of the skin by distinguishing the boundary between the window material of the objective lens and the skin (the beginning of the skin), a signal derived from CH stretching vibration of protein contained in the skin was used as a guide. Details will be described below.
As described above, the signal intensity derived from the CH stretching vibration of the protein of the Raman spectrum shows a maximum value on the skin side slightly from the boundary between the window material of the objective lens and the skin, and then the signal intensity decreases as it goes deeper. Since there is no signal derived from protein CH stretching vibration in the window material, the signal intensity derived from CH stretching vibration is 0 when the focal point of the laser is within the window material. On the other hand, when the signal intensity derived from the CH stretching vibration when the laser is focused in the skin is 100, the signal intensity derived from the CH stretching vibration is 50 when the focus comes to the interface between the window material and the skin. In practice, however, the window material may be detected even though the signal intensity derived from the CH stretching vibration is weak. Therefore, in consideration of the spatial resolution of the nano finder, which is the confocal Raman spectroscopic apparatus used for the measurement in the present invention, the water content at a portion 2 μm deep from the surface, which is the actual measurement value, is defined as “surface water content”. .
The results are shown in Table 3.
表3に示すように、水負荷前後の表面水分量が変化した。具体的には、水負荷量が0.02mL/cm2では、水分負荷後の表面水分量(%)は水分負荷前と比較して1.39倍に増加した。同様に、前述の基準状態とした、水分負荷量が0.04mL/cm2の場合では、水分負荷後の表面水分量(%)は水分負荷前と比較して2.40倍に増加し、水分負荷量が0.20mL/cm2では2.99倍に増加した。
よって、本発明の評価方法における基準値として、皮膚表面の水分量も用いることができる。 As shown in Table 3, the surface moisture content before and after the water load changed. Specifically, when the water load was 0.02 mL / cm 2 , the surface water content (%) after the water load increased 1.39 times compared to that before the water load. Similarly, in the case where the moisture load amount is 0.04 mL / cm 2 in the above-described reference state, the surface moisture amount (%) after the moisture load increases 2.40 times compared to before the moisture load, When the water load was 0.20 mL / cm 2 , it increased 2.99 times.
Therefore, the moisture content on the skin surface can also be used as the reference value in the evaluation method of the present invention.
よって、本発明の評価方法における基準値として、皮膚表面の水分量も用いることができる。 As shown in Table 3, the surface moisture content before and after the water load changed. Specifically, when the water load was 0.02 mL / cm 2 , the surface water content (%) after the water load increased 1.39 times compared to that before the water load. Similarly, in the case where the moisture load amount is 0.04 mL / cm 2 in the above-described reference state, the surface moisture amount (%) after the moisture load increases 2.40 times compared to before the moisture load, When the water load was 0.20 mL / cm 2 , it increased 2.99 times.
Therefore, the moisture content on the skin surface can also be used as the reference value in the evaluation method of the present invention.
さらに、水分負荷前後の、皮膚表面から深さ13μmの初期の角層厚に相当する深さの水分量の変化量(Δ深さ13μmの水分量)についても算出した。その結果を表4に示す。
Furthermore, the amount of change in water content at a depth corresponding to the initial stratum corneum thickness of 13 μm deep before and after the moisture load (Δ 13 μm water content) was also calculated. The results are shown in Table 4.
表4に示すように、負荷する水分量に応じて、水負荷前の角層の厚さに相当する深さ13μmにおける水分量が変化した。水負荷前後の深さ13μmの水分量を比較した場合も物質浸透性が亢進した水負荷量0.04mL/cm2以降で増加し、すなわち、初期の角層厚に相当する部位の深さの水分量が増加したことがわかった。
よって、本発明の評価方法における基準値として、初期の角層厚に相当する深さの部位の水分量の変化量も用いることができる。 As shown in Table 4, the amount of water at a depth of 13 μm corresponding to the thickness of the stratum corneum before the water load changed according to the amount of water to be loaded. When the water content at a depth of 13 μm before and after the water load is compared, the water load increases after the water load of 0.04 mL / cm 2 where the substance permeability is increased, that is, the water at the depth corresponding to the initial stratum corneum thickness. It was found that the amount increased.
Therefore, as the reference value in the evaluation method of the present invention, it is also possible to use the amount of change in the water content at the depth corresponding to the initial stratum corneum thickness.
よって、本発明の評価方法における基準値として、初期の角層厚に相当する深さの部位の水分量の変化量も用いることができる。 As shown in Table 4, the amount of water at a depth of 13 μm corresponding to the thickness of the stratum corneum before the water load changed according to the amount of water to be loaded. When the water content at a depth of 13 μm before and after the water load is compared, the water load increases after the water load of 0.04 mL / cm 2 where the substance permeability is increased, that is, the water at the depth corresponding to the initial stratum corneum thickness. It was found that the amount increased.
Therefore, as the reference value in the evaluation method of the present invention, it is also possible to use the amount of change in the water content at the depth corresponding to the initial stratum corneum thickness.
次に、皮膚表面からの深さ1μmから水負荷前の初期の角層厚に相当する13μmの深さまでの水分量の曲線下面積(Area Under the Curve (AUC) of water profile 1-13μm)の結果を表5に示す。統計解析にはIBM SPSSバージョン23のソフトウェアを使用した。
Next, the area under the curve (Area Under the Curve (AUC) of water profile 1-13µm) from the skin surface to a depth of 13µm corresponding to the initial stratum corneum thickness before water loading The results are shown in Table 5. IBM SPSS version 23 software was used for statistical analysis.
表5に示すように、水負荷前後の皮膚表面からの深さ1μmから13μmまでの深さの水分量の曲線下面積が変化した。具体的には、水負荷量が0.02mL/cm2では、水分負荷後の水分量の曲線下面積は水分負荷前と比較して1.20倍に増加した。同様に、前述の基準状態とした、水分負荷量が0.04mL/cm2の場合では、水分負荷後の水分量の曲線下面積は水分負荷前と比較して1.59倍に増加し、水分負荷量が0.20mL/cm2では1.83倍に増加した。
よって、本発明の評価方法における基準値として、皮膚表面からの所定の深さまでにおける皮膚の水分量の曲線下面積も用いることができる。 As shown in Table 5, the area under the curve of the water content at a depth of 1 μm to 13 μm from the skin surface before and after the water load changed. Specifically, when the water load was 0.02 mL / cm 2 , the area under the curve of the water amount after the water load increased 1.20 times compared with that before the water load. Similarly, in the case where the moisture load amount is 0.04 mL / cm 2 in the above-described reference state, the area under the curve of the moisture amount after the moisture load increases 1.59 times compared to before the moisture load, When the water load was 0.20 mL / cm 2 , it increased 1.83 times.
Therefore, the area under the curve of the moisture content of the skin up to a predetermined depth from the skin surface can also be used as the reference value in the evaluation method of the present invention.
よって、本発明の評価方法における基準値として、皮膚表面からの所定の深さまでにおける皮膚の水分量の曲線下面積も用いることができる。 As shown in Table 5, the area under the curve of the water content at a depth of 1 μm to 13 μm from the skin surface before and after the water load changed. Specifically, when the water load was 0.02 mL / cm 2 , the area under the curve of the water amount after the water load increased 1.20 times compared with that before the water load. Similarly, in the case where the moisture load amount is 0.04 mL / cm 2 in the above-described reference state, the area under the curve of the moisture amount after the moisture load increases 1.59 times compared to before the moisture load, When the water load was 0.20 mL / cm 2 , it increased 1.83 times.
Therefore, the area under the curve of the moisture content of the skin up to a predetermined depth from the skin surface can also be used as the reference value in the evaluation method of the present invention.
表3~5に示す結果より、物質の浸透性が亢進していない水負荷量0.02mL/cm2の場合の表面水分量、深さ13μmの水分量又は水分量の曲線下面積の変化量と比べて、皮膚のバリア機能が低下し、物質浸透性が亢進した水負荷量0.04mL/cm2以降では表面水分量、深さ13μmの水分量又は水分量の曲線下面積の変化量が大きく増加したことで、水溶性物質が分配されやすくなり、より浸透しやすくなった可能性が推察できた。
From the results shown in Tables 3 to 5, the amount of change in the area under the curve of the surface moisture content, the moisture content at a depth of 13 μm, or the moisture content when the water load is 0.02 mL / cm 2 where the permeability of the substance is not enhanced. Compared to the above, the water load of 0.04 mL / cm 2 after the skin barrier function is reduced and the substance permeability is increased, the surface water content, the water content at a depth of 13 μm, or the amount of change in the area under the curve of the water content is It can be inferred that the large increase makes it easier for water-soluble substances to be distributed and easier to penetrate.
以上のように、コルネオメーターやラマン分光法などで非侵襲的に測定した皮膚の水分量や水分分布から、水負荷量がある一定値以上増加すると物質の透過性が亢進することがわかった。すなわち、コルネオメーターやラマン分光法などで非侵襲的に測定した皮膚の水分量や水分分布から、基準状態における皮膚水分量又は水分分布との比較により、物質の透過性、すなわち皮膚のバリア機能を評価することができる。
As described above, it was found from the moisture content and moisture distribution of the skin measured non-invasively with a corneometer, Raman spectroscopy, etc. that the permeability of the substance increases when the water load increases beyond a certain value. That is, the permeability of the substance, that is, the barrier function of the skin, is compared with the skin moisture content or moisture distribution in the reference state from the skin moisture content or moisture distribution measured non-invasively by a corneometer or Raman spectroscopy. Can be evaluated.
試験例2 吸収性物品の性能比較試験
(1)皮膚への水分の負荷
健常な女性1名(30代)を被験者とし、吸収性物品が前腕内側の皮膚へ与える影響を評価した。測定は24~25℃、相対湿度40~50%の環境可変室で、20~30分順化した後に行った。 Test Example 2 Performance Comparison Test of Absorbent Article (1) Moisture load on skin One healthy woman (30's) was the subject, and the effect of the absorbent article on the skin on the inner side of the forearm was evaluated. The measurement was performed after acclimatization for 20 to 30 minutes in an environment variable room at 24 to 25 ° C. and a relative humidity of 40 to 50%.
(1)皮膚への水分の負荷
健常な女性1名(30代)を被験者とし、吸収性物品が前腕内側の皮膚へ与える影響を評価した。測定は24~25℃、相対湿度40~50%の環境可変室で、20~30分順化した後に行った。 Test Example 2 Performance Comparison Test of Absorbent Article (1) Moisture load on skin One healthy woman (30's) was the subject, and the effect of the absorbent article on the skin on the inner side of the forearm was evaluated. The measurement was performed after acclimatization for 20 to 30 minutes in an environment variable room at 24 to 25 ° C. and a relative humidity of 40 to 50%.
(2)コルネオメーターを用いた皮膚水分量の測定
2種類の吸収性物品(吸収性物品A及び吸収性物品B)を5.0cm×5.0cmに切断して試験片を調製し、それぞれに生理食塩水3.0mLを注入し(0.12mL/cm2試験片面積)、それらを前腕内側に3時間貼付した。なお、吸収性物品Aは市販の使い捨ておむつであり、吸収性物品Bは市販の綿製のおむつである。
試験片貼付前後の皮膚水分量を、試験例1と同様コルネオメーターで測定し、試験片貼付前後の皮膚水分量の変化量を算出した。その結果を表6に示す。 (2) Measurement of skin moisture using a corneometer Two types of absorbent articles (absorbent article A and absorbent article B) were cut into 5.0 cm × 5.0 cm to prepare test pieces, respectively. Saline (3.0 mL) was injected (0.12 mL / cm 2 test piece area), and they were affixed to the inner side of the forearm for 3 hours. The absorbent article A is a commercially available disposable diaper, and the absorbent article B is a commercially available cotton diaper.
The skin moisture content before and after applying the test piece was measured with a corneometer in the same manner as in Test Example 1, and the amount of change in the skin moisture content before and after applying the test piece was calculated. The results are shown in Table 6.
2種類の吸収性物品(吸収性物品A及び吸収性物品B)を5.0cm×5.0cmに切断して試験片を調製し、それぞれに生理食塩水3.0mLを注入し(0.12mL/cm2試験片面積)、それらを前腕内側に3時間貼付した。なお、吸収性物品Aは市販の使い捨ておむつであり、吸収性物品Bは市販の綿製のおむつである。
試験片貼付前後の皮膚水分量を、試験例1と同様コルネオメーターで測定し、試験片貼付前後の皮膚水分量の変化量を算出した。その結果を表6に示す。 (2) Measurement of skin moisture using a corneometer Two types of absorbent articles (absorbent article A and absorbent article B) were cut into 5.0 cm × 5.0 cm to prepare test pieces, respectively. Saline (3.0 mL) was injected (0.12 mL / cm 2 test piece area), and they were affixed to the inner side of the forearm for 3 hours. The absorbent article A is a commercially available disposable diaper, and the absorbent article B is a commercially available cotton diaper.
The skin moisture content before and after applying the test piece was measured with a corneometer in the same manner as in Test Example 1, and the amount of change in the skin moisture content before and after applying the test piece was calculated. The results are shown in Table 6.
表6に示すように、吸収性物品Aを貼付した場合と比較して、吸収性物品Bを貼付した場合、皮膚水分量が増加していた。
また、吸収性物品Aを貼付した場合の皮膚水分量の変化量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の皮膚水分量の変化量(19.8±8.6(A.U))よりも少なかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の皮膚水分量の変化量は、前記基準値よりも多いので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 6, when the absorbent article B was affixed, the skin moisture content was increased compared to the case where the absorbent article A was affixed.
The amount of change in skin moisture when the absorbent article A was applied was the reference value in Test Example 1, and the amount of change in skin moisture when the moisture load was 0.04 mL / cm 2 (19.8 ± 8.6 (AU)). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the amount of change in the amount of skin moisture when the absorbent article B is affixed is greater than the reference value, it can be evaluated that the barrier function decreases when the absorbent article B is affixed.
また、吸収性物品Aを貼付した場合の皮膚水分量の変化量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の皮膚水分量の変化量(19.8±8.6(A.U))よりも少なかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の皮膚水分量の変化量は、前記基準値よりも多いので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 6, when the absorbent article B was affixed, the skin moisture content was increased compared to the case where the absorbent article A was affixed.
The amount of change in skin moisture when the absorbent article A was applied was the reference value in Test Example 1, and the amount of change in skin moisture when the moisture load was 0.04 mL / cm 2 (19.8 ± 8.6 (AU)). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the amount of change in the amount of skin moisture when the absorbent article B is affixed is greater than the reference value, it can be evaluated that the barrier function decreases when the absorbent article B is affixed.
(3)共焦点ラマン分光顕微鏡を用いた皮膚水分量の測定
試験例1と同様に、皮膚表面から40μmまで、深さ方向に2μm間隔毎にラマンスペクトルを測定した。このようにして測定し特開2012-50739号公報に記載の方法に従って水分量を近似的に算出し、深さ方向の皮膚内の水分量分布の値をグラフにプロットした。その結果を図3に示す。 (3) Measurement of skin water content using confocal Raman spectroscopic microscope In the same manner as in Test Example 1, a Raman spectrum was measured every 2 μm in the depth direction from the skin surface to 40 μm. The water content was measured and measured in accordance with the method described in JP2012-50739A, and the water content distribution value in the skin in the depth direction was plotted on a graph. The result is shown in FIG.
試験例1と同様に、皮膚表面から40μmまで、深さ方向に2μm間隔毎にラマンスペクトルを測定した。このようにして測定し特開2012-50739号公報に記載の方法に従って水分量を近似的に算出し、深さ方向の皮膚内の水分量分布の値をグラフにプロットした。その結果を図3に示す。 (3) Measurement of skin water content using confocal Raman spectroscopic microscope In the same manner as in Test Example 1, a Raman spectrum was measured every 2 μm in the depth direction from the skin surface to 40 μm. The water content was measured and measured in accordance with the method described in JP2012-50739A, and the water content distribution value in the skin in the depth direction was plotted on a graph. The result is shown in FIG.
図3に示すように、吸収性物品Aを貼付した場合と比較して、吸収性物品Bを貼付した場合、皮膚水分量が増加していた。また同様に、吸収性物品Aを貼付した場合と比較して、吸収性物品Bを貼付した場合、水分負荷前の初期の角層厚よりも深いところの水分量が増加した。
よって、水分分布を用いて初期の角層厚よりも深部の水分量の差からも、吸収性物品Aを貼付してもバリア機能は変化しないのに対し、吸収性物品Bを貼付するとバリア機能は低下する、と評価できる。 As shown in FIG. 3, compared with the case where the absorbent article A was affixed, the amount of skin moisture was increased when the absorbent article B was affixed. Similarly, when the absorbent article B is affixed, the amount of water deeper than the initial stratum corneum thickness before the moisture load is increased as compared with the case where the absorbent article A is affixed.
Therefore, the barrier function does not change even if the absorbent article A is pasted from the difference in the moisture content deeper than the initial stratum corneum thickness using the moisture distribution, whereas the barrier function when the absorbent article B is pasted Can be evaluated as decreasing.
よって、水分分布を用いて初期の角層厚よりも深部の水分量の差からも、吸収性物品Aを貼付してもバリア機能は変化しないのに対し、吸収性物品Bを貼付するとバリア機能は低下する、と評価できる。 As shown in FIG. 3, compared with the case where the absorbent article A was affixed, the amount of skin moisture was increased when the absorbent article B was affixed. Similarly, when the absorbent article B is affixed, the amount of water deeper than the initial stratum corneum thickness before the moisture load is increased as compared with the case where the absorbent article A is affixed.
Therefore, the barrier function does not change even if the absorbent article A is pasted from the difference in the moisture content deeper than the initial stratum corneum thickness using the moisture distribution, whereas the barrier function when the absorbent article B is pasted Can be evaluated as decreasing.
さらに、図3に示す水分プロファイルから、表面水分量(皮膚表面からの深さ2μmの部位の水分量)を算出した。その結果を表7に示す。
Furthermore, from the moisture profile shown in FIG. 3, the surface moisture content (moisture content at a site having a depth of 2 μm from the skin surface) was calculated. The results are shown in Table 7.
表7に示すように、水負荷前後の表面水分量(%)は吸収性物品Aの貼付後は貼付前と比較して、1.19倍増加した。これに対して吸収性物品Bの貼付後は貼付前と比較して、2.86倍増加した。
また、吸収性物品Aを貼付した場合の表面水分量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の表面水分量の増加分(2.40倍)よりも小さかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の表面水分量の増加分は、前記基準値よりも大きいので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 7, the surface moisture content (%) before and after the water load increased 1.19 times after application of the absorbent article A compared to before application. On the other hand, it increased 2.86 times after sticking of the absorbent article B compared with before sticking.
Further, the surface moisture content when the absorbent article A was applied was the reference value in Test Example 1, and the increase in the surface moisture content when the moisture load was 0.04 mL / cm 2 (2.40 times). Was smaller than. Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the increase in the amount of surface moisture when the absorbent article B is affixed is greater than the reference value, it can be evaluated that the barrier function decreases when the absorptive article B is affixed.
また、吸収性物品Aを貼付した場合の表面水分量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の表面水分量の増加分(2.40倍)よりも小さかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の表面水分量の増加分は、前記基準値よりも大きいので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 7, the surface moisture content (%) before and after the water load increased 1.19 times after application of the absorbent article A compared to before application. On the other hand, it increased 2.86 times after sticking of the absorbent article B compared with before sticking.
Further, the surface moisture content when the absorbent article A was applied was the reference value in Test Example 1, and the increase in the surface moisture content when the moisture load was 0.04 mL / cm 2 (2.40 times). Was smaller than. Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the increase in the amount of surface moisture when the absorbent article B is affixed is greater than the reference value, it can be evaluated that the barrier function decreases when the absorptive article B is affixed.
次に、図3に示す水分プロファイルから、皮膚表面から深さ13μmの初期の角層厚に相当する深さの水分量の変化量(Δ深さ13μmの水分量)を算出した。その結果を表8に示す。
Next, from the moisture profile shown in FIG. 3, the amount of change in the moisture content at a depth corresponding to the initial stratum corneum thickness of 13 μm deep from the skin surface (Δ 13 μm moisture content) was calculated. The results are shown in Table 8.
表8に示すように、吸収性物品Aを貼付した場合と比較して、吸収性物品Bを貼付した場合、深さ13μmにおける水分量が大きく増加した。
また、吸収性物品Aを貼付した場合の初期の角層厚に相当する深さの水分量の変化量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の初期の角層厚に相当する深さの水分量の変化量(9.21±9.2(%))よりも少なかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の初期の角層厚に相当する深さの水分量の変化量は、前記基準値よりも多いので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 8, when the absorbent article B was affixed, the water content at a depth of 13 μm was greatly increased as compared to the case where the absorbent article A was affixed.
In addition, when the absorbent article A is pasted, the amount of change in the water content at the depth corresponding to the initial stratum corneum thickness is the reference value in Test Example 1, and the water load is 0.04 mL / cm 2 It was less than the amount of change in water content at the depth corresponding to the initial stratum corneum thickness (9.21 ± 9.2 (%)). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the amount of change in the moisture content at the depth corresponding to the initial stratum corneum thickness when the absorbent article B is affixed is larger than the reference value, it is evaluated that the barrier function decreases when the absorptive article B is affixed. it can.
また、吸収性物品Aを貼付した場合の初期の角層厚に相当する深さの水分量の変化量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の初期の角層厚に相当する深さの水分量の変化量(9.21±9.2(%))よりも少なかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の初期の角層厚に相当する深さの水分量の変化量は、前記基準値よりも多いので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 8, when the absorbent article B was affixed, the water content at a depth of 13 μm was greatly increased as compared to the case where the absorbent article A was affixed.
In addition, when the absorbent article A is pasted, the amount of change in the water content at the depth corresponding to the initial stratum corneum thickness is the reference value in Test Example 1, and the water load is 0.04 mL / cm 2 It was less than the amount of change in water content at the depth corresponding to the initial stratum corneum thickness (9.21 ± 9.2 (%)). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the amount of change in the moisture content at the depth corresponding to the initial stratum corneum thickness when the absorbent article B is affixed is larger than the reference value, it is evaluated that the barrier function decreases when the absorptive article B is affixed. it can.
さらに、図3に示す水分分布から、試験片貼付前後の皮膚表面からの深さ1μmから13μmの深さまでの水分量の曲線下面積(AUC of water profile 1-13μm)を算出した。その結果を表9に示す。
Furthermore, from the moisture distribution shown in FIG. 3, the area under the curve (AUC of water profile 1-13 μm) of the moisture content from the depth of 1 μm to the depth of 13 μm from the skin surface before and after applying the test piece was calculated. The results are shown in Table 9.
表9に示すように、水負荷前後の皮膚表面からの深さ1μmから13μmまでの深さの水分量の曲線下面積は吸収性物品Aの貼付後は貼付前と比較して、1.08倍増加した。これに対して吸収性物品Bの貼付後は貼付前と比較して、1.89倍増加した。
また、吸収性物品Aを貼付した場合の曲線下面積の変化量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の曲線下面積の増加分(1.59倍)よりも小さかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の曲線下面積の増加分は前記基準値よりも大きいので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 9, the area under the curve of the water content at a depth of 1 μm to 13 μm from the skin surface before and after water load is 1.08 after the application of the absorbent article A and before the application. Doubled. On the other hand, after application of the absorbent article B, the increase was 1.89 times as compared with before application.
In addition, the amount of change in the area under the curve when the absorbent article A was applied was the reference value in Test Example 1, and the increase in the area under the curve when the moisture load was 0.04 mL / cm 2 (1. 59 times). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the increase in the area under the curve when the absorbent article B is pasted is larger than the reference value, it can be evaluated that the barrier function is lowered when the absorbent article B is pasted.
また、吸収性物品Aを貼付した場合の曲線下面積の変化量は、試験例1において基準値とした、水分負荷量が0.04mL/cm2の場合の曲線下面積の増加分(1.59倍)よりも小さかった。よって、吸収性物品Aを貼付しても、バリア機能は変化していないと評価できる。一方、吸収性物品Bを貼付した場合の曲線下面積の増加分は前記基準値よりも大きいので、吸収性物品Bを貼付するとバリア機能は低下すると評価できる。 As shown in Table 9, the area under the curve of the water content at a depth of 1 μm to 13 μm from the skin surface before and after water load is 1.08 after the application of the absorbent article A and before the application. Doubled. On the other hand, after application of the absorbent article B, the increase was 1.89 times as compared with before application.
In addition, the amount of change in the area under the curve when the absorbent article A was applied was the reference value in Test Example 1, and the increase in the area under the curve when the moisture load was 0.04 mL / cm 2 (1. 59 times). Therefore, even if the absorbent article A is pasted, it can be evaluated that the barrier function is not changed. On the other hand, since the increase in the area under the curve when the absorbent article B is pasted is larger than the reference value, it can be evaluated that the barrier function is lowered when the absorbent article B is pasted.
(4)物質透過性の評価
試験片を除去した直後に、0.007質量%の濃度のニコチン酸メチル水溶液を15秒、試験片を貼付した皮膚に負荷し、ニコチン酸メチル処理から20分後の紅斑スコアを試験例1と同様にして目視判定した。
その結果を表10に示す。 (4) Evaluation of substance permeability Immediately after removing the test piece, a 0.007 mass% concentration of methyl nicotinate aqueous solution was applied to the skin on which the test piece was applied for 15 seconds, and 20 minutes after treatment with methyl nicotinate. The erythema score was visually determined in the same manner as in Test Example 1.
The results are shown in Table 10.
試験片を除去した直後に、0.007質量%の濃度のニコチン酸メチル水溶液を15秒、試験片を貼付した皮膚に負荷し、ニコチン酸メチル処理から20分後の紅斑スコアを試験例1と同様にして目視判定した。
その結果を表10に示す。 (4) Evaluation of substance permeability Immediately after removing the test piece, a 0.007 mass% concentration of methyl nicotinate aqueous solution was applied to the skin on which the test piece was applied for 15 seconds, and 20 minutes after treatment with methyl nicotinate. The erythema score was visually determined in the same manner as in Test Example 1.
The results are shown in Table 10.
表10に示すように、吸収性物品Aを貼付した場合と比較して、吸収性物品Bを貼付した場合紅斑スコアが上昇し、ニコチン酸メチルの透過性が亢進していた。
As shown in Table 10, when the absorbent article B was affixed, the erythema score increased and the permeability of methyl nicotinate was enhanced as compared with the case where the absorbent article A was affixed.
上記結果に示すように、吸収性物品Aを貼付した場合、コルネオメーターによる皮膚の水分量の増加、並びにラマン分光法で測定した皮膚の表面水分量の増加、深さ13μmにおける水分量の増加、及び皮膚表面からの深さ1μmから13μmまでの水分量の曲線下面積の増加はいずれも、基準とする水分分布に比べて小さいと評価できた。よって、皮膚の水分量吸収性物品Aを貼付した場合は、水溶性物質の透過性が低いと判断される。吸収性物品Aを貼付して水溶性物質の透過性を実際に判定したところ、皮膚の外側から内側への水溶性物質の透過性が低かった。すなわち吸収性物品Aは、上記結果から、水溶性物質の皮膚への透過性は亢進されず、皮膚トラブルを起こしにくい吸収性物品であると言える。
As shown in the above results, when the absorbent article A was affixed, an increase in the amount of moisture in the skin with a corneometer, an increase in the amount of moisture on the skin as measured by Raman spectroscopy, an increase in the amount of moisture at a depth of 13 μm, In addition, the increase in the area under the curve of the water content from the depth of 1 μm to 13 μm from the skin surface could be evaluated to be smaller than the reference water distribution. Therefore, when skin moisture-absorbing article A is affixed, it is determined that the permeability of the water-soluble substance is low. When the absorbent article A was applied and the permeability of the water-soluble substance was actually judged, the permeability of the water-soluble substance from the outside to the inside of the skin was low. That is, from the above results, it can be said that the absorbent article A is an absorbent article that does not increase the permeability of the water-soluble substance to the skin and hardly causes skin troubles.
一方、吸収性物品Bを貼付した場合、コルネオメーターによる皮膚の水分量の増加、並びにラマン分光法で測定した皮膚の表面水分量の増加、深さ13μmにおける水分量の増加、及び皮膚表面からの深さ1μmから13μmまでの水分量の曲線下面積の増加はいずれも、基準とする水分分布に比べて大きいと評価できた。よって、吸収性物品Bを貼付した場合は、水溶性物質の透過性が高いと判断される。吸収性物品Bを貼付して水溶性物質の透過性を実際に判定したところ、驚くべきことに皮膚の外側から内側への水溶性物質の透過性が上昇していたことがわかった。すなわち吸収性物品Bは、水溶性物質の皮膚への透過性が亢進され、皮膚トラブルを起こす可能性がある吸収性物品であると言える。
On the other hand, when the absorbent article B is affixed, an increase in the amount of moisture in the skin by a corneometer, an increase in the amount of moisture on the skin measured by Raman spectroscopy, an increase in the amount of moisture at a depth of 13 μm, and It was possible to evaluate that the increase in the area under the curve of the water content from the depth of 1 μm to 13 μm was larger than the reference water distribution. Therefore, when the absorbent article B is affixed, it is determined that the water-soluble substance has high permeability. When the absorbent article B was affixed and the permeability of the water-soluble substance was actually judged, it was surprisingly found that the permeability of the water-soluble substance from the outside to the inside of the skin was increased. That is, it can be said that the absorbent article B is an absorbent article in which the permeability of the water-soluble substance to the skin is enhanced and skin trouble may occur.
以上より、本発明の方法に従って被験物質又は物品をヒトの皮膚に接触させ、接触前後の皮膚の水分量又は水分分布を測定した結果から、前記被験物質又は物品が、皮膚のバリア機能に与える影響を評価する方法として有効であることが確認できた。
As described above, the test substance or article is brought into contact with human skin according to the method of the present invention, and the effect of the test substance or article on the skin barrier function is determined from the result of measuring the moisture content or moisture distribution of the skin before and after contact. It was confirmed that it is effective as a method for evaluating
本発明をその実施態様とともに説明したが、我々は特に指定しない限り我々の発明を説明のどの細部においても限定しようとするものではなく、添付の請求の範囲に示した発明の精神と範囲に反することなく幅広く解釈されるべきであると考える。
While this invention has been described in conjunction with its embodiments, we do not intend to limit our invention in any detail of the description unless otherwise specified and are contrary to the spirit and scope of the invention as set forth in the appended claims. I think it should be interpreted widely.
Claims (29)
- 被験物質又は物品をヒトの皮膚に接触させ、
接触前後の皮膚の水分量又は水分分布の変化量を、基準値と比較し、
比較結果から、前記被験物質又は物品が、皮膚バリア機能に与える影響を評価する、
被験物質又は物品の評価方法であって、
水の負荷量を変えての皮膚への水負荷試験により測定された、皮膚の水分量又は水分分布と、皮膚のバリア機能とが関連付けられ、
皮膚の水分量又は水分分布と皮膚のバリア機能との関連性から、皮膚のバリア機能に変化が確認されるときの皮膚の水分量又は水分分布が、前記基準値として設定されたものである、
被験物質又は物品の評価方法。 Bringing the test substance or article into contact with human skin;
Compare the amount of moisture or moisture distribution in the skin before and after contact with the reference value,
From the comparison results, the effect of the test substance or article on the skin barrier function is evaluated.
A method for evaluating a test substance or article,
The water content or distribution of the skin, measured by a water load test on the skin with different water loads, is associated with the barrier function of the skin,
From the relationship between the moisture content or moisture distribution of the skin and the barrier function of the skin, the moisture content or moisture distribution of the skin when the change is confirmed in the skin barrier function is set as the reference value.
Evaluation method of test substance or article. - 皮膚のバリア機能の変化を確認するために、皮膚外部から内部への指標物質の浸透性がさらに測定され、指標物質の浸透性の結果から前記基準値が設定されたものである、請求項1に記載の方法。 In order to confirm the change in the barrier function of the skin, the permeability of the indicator substance from the outside to the inside of the skin is further measured, and the reference value is set from the result of the permeability of the indicator substance. The method described in 1.
- 前記被験物質又は物品の接触後の皮膚水分量が、基準値以上の変化量で増加した場合、バリア機能が低下したと評価する、請求項1又は2に記載の方法。 The method according to claim 1 or 2, wherein when the amount of skin moisture after contact with the test substance or article increases with a change amount greater than or equal to a reference value, the barrier function is evaluated to be lowered.
- 前記被験物質又は物品の接触後の皮膚水分量の変化量が、基準値より小さい場合、バリア機能は変化していないと評価する、請求項1~3のいずれか1項に記載の方法。 The method according to any one of claims 1 to 3, wherein when the amount of change in skin moisture after contact with the test substance or article is smaller than a reference value, the barrier function is evaluated not to change.
- 被験物質又は物品をヒトの皮膚に接触させた後のヒトの皮膚の深さごとの水分量を測定して、被験物質又は物品の接触後の水分分布を取得し、
被験物質又は物品の接触後の水分分布と基準水分分布とを比較することで、被験物質又は物品が、ヒトの皮膚のバリア機能に与える影響を評価する、被験物質又は物品の評価方法であって、
水又は水溶液のヒトの皮膚への負荷前に、ヒトの皮膚の深さ毎の水分量を測定して初期水分分布を取得し、
水又は水溶液のヒトの皮膚への負荷終了直後に、ヒトの皮膚の深さ毎の水分量の測定による水又は水溶液の負荷後の水分分布の取得と、指標物質を皮膚に適用して該指標物質の浸透度の測定を行い、
初期水分分布から水又は水溶液の負荷終了直後の水分分布への変化と、指標物質の浸透度から、指標物質の浸透度と水又は水溶液の負荷量との関連性を決定し、
前記関連性が変化したときのヒトの皮膚の深さ毎の水又は水溶液の負荷後の水分分布を、前記基準水分分布とする、
被験物質又は物品の評価方法。 Measure the amount of water per depth of human skin after contacting the test substance or article with human skin to obtain the moisture distribution after contact with the test substance or article,
A test substance or article evaluation method for evaluating the influence of a test substance or article on the barrier function of human skin by comparing the moisture distribution after contact of the test substance or article with a reference moisture distribution. ,
Before loading water or aqueous solution on human skin, measure the amount of water for each depth of human skin to obtain the initial moisture distribution,
Immediately after the end of loading of water or aqueous solution on human skin, acquisition of water distribution after loading water or aqueous solution by measuring the amount of water for each depth of human skin, and applying the indicator substance to the skin, the indicator Measure the permeability of the substance,
From the initial moisture distribution to the moisture distribution immediately after the end of the load of water or aqueous solution, and the permeability of the indicator substance, determine the relationship between the permeability of the indicator substance and the load of water or aqueous solution,
The water distribution after loading with water or an aqueous solution at every depth of the human skin when the relevance is changed as the reference water distribution,
Evaluation method of test substance or article. - 被験物質又は物品の接触前と接触後のヒトの皮膚の所定の深さ位置の水分量を測定し、接触前の水分量と接触後の水分量との差を取得し、
取得した水分量の差と、前記基準水分分布とに基づき、皮膚バリア機能に与える影響を評価する、
請求項5に記載の評価方法。 Measure the moisture content at a predetermined depth of human skin before and after contact with the test substance or article, and obtain the difference between the moisture content before contact and the moisture content after contact,
Based on the obtained moisture difference and the reference moisture distribution, to evaluate the effect on the skin barrier function,
The evaluation method according to claim 5. - 前記指標物質が、ニコチン酸メチル、ニコチン酸ヘキシル、乳酸、黄色4号、赤色215号、カフェイン、ユビナール、カロテノイド、レチノール、並びに、皮膚保湿剤、皮膚外用剤、皮膚化粧料若しくは蒸気温熱具に含まれる有効成分若しくは薬効成分からなる群より選ばれる少なくとも1種である、請求項2~6のいずれか1項に記載の方法。 The indicator substance is methyl nicotinate, hexyl nicotinate, lactic acid, yellow No. 4, red No. 215, caffeine, ubinal, carotenoid, retinol, and skin moisturizer, skin external preparation, skin cosmetic, or steam heating device. The method according to any one of claims 2 to 6, which is at least one selected from the group consisting of an active ingredient or a medicinal ingredient.
- 皮膚の水分量を皮膚静電容量測定手段により測定し解析する、請求項1~7のいずれか1項に記載の方法。 The method according to any one of claims 1 to 7, wherein the moisture content of the skin is measured and analyzed by a skin capacitance measuring means.
- 健常者の皮膚を評価対象とし、被験物質又は物品の接触後の皮膚の水分量を皮膚静電容量測定手段により測定し、被験物質又は物品の接触後の皮膚の水分量が、被験物質又は物品の接触前の皮膚の水分量に対して、8(A.U.)以上の変化量で増加した場合、バリア機能は低下したと評価する、請求項8に記載の方法。 Measure skin moisture after contact with the test substance or article with the skin capacitance measuring means, and the skin moisture after contact with the test substance or article is the test substance or article. The method according to claim 8, wherein the barrier function is evaluated to be reduced when the moisture content of the skin before contact increases by a change amount of 8 (AU) or more.
- 健常者の皮膚を評価対象とし、被験物質又は物品の接触後の皮膚の水分量を皮膚静電容量測定手段により測定し、被験物質又は物品の接触後の皮膚の水分量が、被験物質又は物品の接触前の皮膚の水分量に対して、16(A.U.)増加より小さい変化量の場合、バリア機能は変化していないと評価する、請求項8に記載の方法。 Measure skin moisture after contact with the test substance or article with the skin capacitance measuring means, and the skin moisture after contact with the test substance or article is the test substance or article. The method according to claim 8, wherein the barrier function is evaluated as not changing when the amount of change is smaller than an increase of 16 (AU) with respect to the amount of moisture in the skin before contact.
- 皮膚表面からの深さ2~40μmの深さまでの水分量又は水分分布をラマン分光法により測定し解析する、請求項1~7のいずれか1項に記載の方法。 The method according to any one of claims 1 to 7, wherein the moisture content or moisture distribution from the skin surface to a depth of 2 to 40 µm is measured and analyzed by Raman spectroscopy.
- 健常者の皮膚を評価対象とし、皮膚表面から所定の深さまでにおける、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合、バリア機能が低下した、と評価する、請求項11に記載の方法。 The skin moisture distribution before and after contact with the test substance or article is measured by Raman spectroscopy from the skin surface to a predetermined depth from the skin surface, and the moisture distribution of the skin before and after contact with the test substance or article is evaluated. The method according to claim 11, wherein the barrier function is evaluated to be lowered when the skin moisture content in the deeper part than the initial stratum corneum thickness is increased by contact with the test substance or article.
- 健常者の皮膚を評価対象とし、ラマン分光法より得られた被験物質又は物品の接触後の皮膚表面水分量が、被験物質又は物品の接触前の皮膚表面水分量に対して、2.0倍以上に増加した場合、バリア機能は低下したと評価する、請求項11に記載の方法。 The skin surface moisture after contact of the test substance or article obtained by Raman spectroscopy is 2.0 times the skin surface moisture before contact of the test substance or article. The method according to claim 11, wherein the barrier function is evaluated to be lowered when the increase is made.
- 健常者の皮膚を評価対象とし、ラマン分光法より得られた被験物質又は物品の接触後の皮膚表面水分量が、被験物質又は物品の接触前の皮膚表面水分量に対して、3.0倍増加より小さい場合、バリア機能は変化していないと評価する、請求項11に記載の方法。 The skin surface moisture after contact of the test substance or article obtained by Raman spectroscopy is 3.0 times the skin surface moisture before contact of the test substance or article. 12. The method of claim 11, wherein if the increase is less than the increase, the barrier function is evaluated as not changing.
- 健常者の皮膚を評価対象とし、ラマン分光法より得られた、被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、被験物質又は物品の接触前の初期の角層厚に相当する部位の水分量に対して、8%以上増加した場合、バリア機能は低下したと評価する、請求項11に記載の方法。 The amount of water at the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the initial angle before contact with the test substance or article. The method according to claim 11, wherein the barrier function is evaluated to be lowered when the water content in the region corresponding to the layer thickness increases by 8% or more.
- 健常者の皮膚を評価対象とし、ラマン分光法より得られた、被験物質又は物品の接触後の初期の角層厚に相当する部位の水分量が、被験物質又は物品の接触前の初期の角層厚に相当する部位の水分量に対して、14%増加より小さい場合、バリア機能は変化していないと評価する、請求項11に記載の方法。 The amount of water at the site corresponding to the initial stratum corneum thickness after contact with the test substance or article obtained by Raman spectroscopy is the initial angle before contact with the test substance or article. The method according to claim 11, wherein the barrier function is evaluated as not changing when the water content in the region corresponding to the layer thickness is smaller than an increase of 14%.
- 健常者の皮膚を評価対象とし、ラマン分光法より得られた、皮膚表面からの所定の深さまでにおける、被験物質又は物品の接触後の皮膚の水分量の曲線下面積を求め、被験物質又は物品の接触後の皮膚の水分量の曲線下面積が、被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対して1.4倍以上に増加した場合、バリア機能は低下したと評価する、請求項11に記載の方法。 Determine the area under the curve of the moisture content of the skin after contact with the test substance or article up to a predetermined depth from the skin surface, obtained from Raman spectroscopy, using the skin of a healthy person as the evaluation target, and the test substance or article When the area under the curve of the moisture content of the skin after contact with the test substance or article increased more than 1.4 times the area under the curve of the moisture content of the skin before the contact with the test substance or article, the barrier function was evaluated to be reduced. The method according to claim 11.
- 健常者の皮膚を評価対象とし、ラマン分光法より得られた、皮膚表面からの所定の深さまでにおける、被験物質又は物品の接触後の皮膚の水分量の曲線下面積を求め、被験物質又は物品の接触後の皮膚の水分量の曲線下面積が、被験物質又は物品の接触前の皮膚の水分量の曲線下面積に対して1.8倍より小さい場合、バリア機能は変化していないと評価する、請求項11に記載の方法。 Determine the area under the curve of the moisture content of the skin after contact with the test substance or article up to a predetermined depth from the skin surface, obtained from Raman spectroscopy, using the skin of a healthy person as the evaluation target, and the test substance or article When the area under the curve of the moisture content of the skin after contact is less than 1.8 times the area under the curve of the moisture content of the skin before contact with the test substance or article, the barrier function is evaluated as unchanged. The method according to claim 11.
- 被験物質又は物品が、ヒトの皮膚のバリア機能に与える影響を評価する方法であって、
被験物質又は物品のヒトの皮膚への接触前と接触後の皮膚の深さ毎の水分量を測定して水分分布を取得し、
接触前と接触後の水分分布における、接触前の初期の角層厚よりも深部の水分量の差を指標として、被験物質又は物品がヒトの皮膚のバリア機能に与える影響を評価する、
被験物質又は物品の評価方法。 A method for evaluating the effect of a test substance or article on the barrier function of human skin,
Obtain water distribution by measuring the amount of water at each skin depth before and after contact of the test substance or article with human skin,
Evaluate the effect of the test substance or article on the barrier function of human skin, using the difference in water content deeper than the initial stratum corneum thickness before contact in the moisture distribution before and after contact,
Evaluation method of test substance or article. - 皮膚表面からの深さ2~40μmの深さまでの水分量をラマン分光法により測定し解析する、請求項19に記載の方法。 20. The method according to claim 19, wherein the moisture content from the skin surface to a depth of 2 to 40 μm is measured and analyzed by Raman spectroscopy.
- 健常者の皮膚を評価対象とし、皮膚表面から所定の深さまでにおける、被験物質又は物品の接触前後の皮膚の水分分布をラマン分光法により測定し、被験物質又は物品の接触前後の皮膚の水分分布を比較し、被験物質又は物品の接触により、初期の角層厚より深部における皮膚の水分量が増加した場合、バリア機能が低下した、と評価する、請求項19又は20に記載の方法。 The skin moisture distribution before and after contact with the test substance or article is measured by Raman spectroscopy from the skin surface to a predetermined depth from the skin surface, and the moisture distribution of the skin before and after contact with the test substance or article is evaluated. 21. The method according to claim 19 or 20, wherein the barrier function is evaluated to be lowered when the amount of moisture in the skin at a deeper depth than the initial stratum corneum thickness is increased by contact with a test substance or an article.
- 皮膚水分量又は水分分布を非侵襲的に測定する、請求項1~21のいずれか1項に記載の方法。 The method according to any one of claims 1 to 21, wherein the skin moisture content or moisture distribution is measured noninvasively.
- 被験物質又は物品を負荷していない状態で正常なバリア機能を有するヒト(健常者)の皮膚で評価する、請求項1~22のいずれか1項に記載の方法。 The method according to any one of claims 1 to 22, wherein the evaluation is performed on the skin of a human (healthy person) having a normal barrier function in a state where the test substance or article is not loaded.
- ラマン分光法、皮膚静電容量測定手段、高周波電流測定手段、赤外分光法、及び近赤外分光法からなる群より選ばれる少なくとも1つに従い皮膚の水分量を測定する、請求項1~7、19、22及び23のいずれか1項に記載の方法。 The moisture content of the skin is measured according to at least one selected from the group consisting of Raman spectroscopy, skin capacitance measurement means, high-frequency current measurement means, infrared spectroscopy, and near infrared spectroscopy. , 19, 22 and 23.
- 前記被験物質が、水若しくは水蒸気、皮膚化粧料、皮膚保湿剤若しくは皮膚外用剤、蒸気温熱具に含まれる有効成分若しくは薬効成分、生理食塩水、リン酸緩衝生理食塩水、並びに生体由来の分泌物若しくは***物からなる群より選ばれる少なくとも1種である、請求項1~24のいずれか1項に記載の方法。 The test substance is water or water vapor, skin cosmetics, skin moisturizer or external preparation for skin, active ingredient or medicinal ingredient contained in steam heating device, physiological saline, phosphate buffered saline, and biologically derived secretions Alternatively, the method according to any one of claims 1 to 24, wherein the method is at least one selected from the group consisting of excreta.
- 前記物品が、吸収性物品、蒸気温熱具、創傷保護材、マスク、又はゴム手袋である、請求項1~24のいずれか1項に記載の方法。 The method according to any one of claims 1 to 24, wherein the article is an absorbent article, a steam heating device, a wound protection material, a mask, or a rubber glove.
- 前記吸収性物品が生理食塩水又は擬似尿などを含有する、請求項26に記載の方法。 The method according to claim 26, wherein the absorbent article contains physiological saline or simulated urine.
- 前記被験物質又は物品をヒトの皮膚に接触させる時間が、1時間以上6時間以下である、請求項1~27のいずれか1項に記載の方法。 The method according to any one of claims 1 to 27, wherein the time for contacting the test substance or article with human skin is 1 hour or more and 6 hours or less.
- 被験物質又は物品のヒトの皮膚への接触による皮膚の水分量又は水分分布の変化量と皮膚のバリア機能との関連性に関する情報を格納し、関連性に関する情報に基づき、被験物質又は物品の接触後の皮膚の水分量又は水分分布から、被験物質又は物品が、皮膚のバリア機能に与える影響を評価するためのシステム。 Stores information on the relationship between the amount of moisture in the skin or the distribution of moisture distribution due to contact of the test substance or article with human skin and the barrier function of the skin, and based on the information on the relationship, contact of the test substance or article A system for evaluating the influence of a test substance or article on the barrier function of the skin from the moisture content or moisture distribution of the later skin.
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