JP4990010B2 - Skin quality judgment method - Google Patents

Description

  The present invention relates to a skin quality determination method related to the skin quality of a subject.

  Skin and scalp conditions such as sensitive skin, dry skin, oily skin, skin with poor moisture retention, skin with poor barrier function, skin that is susceptible to acne, skin that is prone to scaling, skin that is prone to erythema, or psoriasis In order to improve the skin condition caused by atopic dermatitis, an appropriate skin quality improving agent needs to be prescribed according to the skin condition. Conventionally, as skin quality improving agents, for example, various steroid preparations, vitamin D3 preparations, vitamin A derivative preparations and other psoriasis treatment agents and skin disease treatment agents such as atopic dermatitis treatment agents are known. Further, as skin quality improving agents, compositions classified into cosmetics such as a moisturizing composition and a whitening composition are also known.

  Conventionally, methods for evaluating skin quality include a method using the abundance of interleukin and interleukin 1 receptor antagonist in the stratum corneum as an index (Patent Document 1), and nerve growth factor (NGF) in the skin stratum corneum. And a method using the abundance of neurotrophic factor (NT-4) as an index is known (Patent Document 2).

  On the other hand, lipids in the stratum corneum are known to be involved in the skin barrier function and water retention function, and are known to be components that greatly affect the skin quality. Here, the lipid refers to a biological molecule having a long-chain fatty acid or a hydrocarbon chain, and includes fatty acid, glyceride, wax ester, sphingolipid, phospholipid, cholesterol and the like. Among them, ceramide, which is a kind of sphingolipid, is a lipid that is greatly involved in these functions, and it has been suggested that the reduction of certain ceramides is related to skin problems such as atopic dermatitis.

Japanese Patent Laid-Open No. 10-216106 JP 2004-28698 A

  However, little is known about what changes in lipid components contained in the skin stratum corneum are related to skin quality and skin-related diseases, and knowledge relating the changes in skin quality and lipid components is known. It is not done. Then, this invention aims at providing the skin quality determination method which can determine the skin quality of a test subject with high precision by clarifying the relationship between various skin quality and the fluctuation | variation of a lipid component. To do.

  As a result of intensive studies by the present inventors in order to achieve the above-described object, the specific skin quality is related to lipid composition information indicating the abundance and abundance ratio of lipid components, in other words, specific skin The quality was found to show characteristic lipid composition information. Then, it was confirmed that the skin quality of the subject can be evaluated and determined by utilizing new knowledge such as the relationship between the skin quality and the lipid composition information, and the present invention has been completed.

  That is, the skin quality determination method according to the present invention is based on lipid composition information obtained from a subject, information on lipid components whose composition ratio changes in relation to skin quality, lipid composition information obtained from a healthy individual, and specific information The skin quality of the subject is determined by comparison with a reference value calculated based on the lipid composition information obtained from the individual with skin quality. Here, examples of the lipid component to be compared include sphingolipids, particularly ceramide.

  In the skin quality determination method according to the present invention, the lipid composition information is obtained by separating and detecting a lipid sample collected from the subject by liquid chromatography-mass spectrometry and analyzing the detection data by a multistage mass chromatogram. It is preferable that it is the data produced | generated by this.

  In the skin quality determination method according to the present invention, skin quality refers to psoriasis, atopic dermatitis, sensitive skin, dry skin, oily skin, skin with poor moisture retention, skin with poor barrier function, and skin that is susceptible to acne. Examples of the skin condition include scalp such as skin that tends to cause scaling and skin that tends to cause erythema. That is, in the skin quality determination method according to the present invention, as information on the lipid component whose composition ratio changes in relation to the skin quality, the information on the lipid component correlated with the skin disease, the skin quality, or the skin condition as described above. Preferably there is. For example, psoriasis and atopic dermatitis can be mentioned as the skin disease, skin quality or skin condition.

  In the case of psoriasis as the above skin disease, skin quality or skin condition, N-acyl-sphingosine / ceramide component (for example, 15% by weight as a reference value), N-acyl-phytosphingosine / ceramide component (for example, 12% as a reference value) %) And N- (α-OH-acyl) -sphingosine / ceramide component (reference value, for example, 10% by weight), the composition ratio of ceramide to the total amount of at least one component selected from the group By doing so, it is possible to determine psoriasis (trend and severity of injury; the same applies hereinafter).

  In the case of psoriasis as the above skin disease, skin quality or skin condition, N-acyl-dihydrosphingosine / ceramide components (for example, 45 as reference values), N-acyl-sphingosine / ceramide components (for example, 44 as reference values) ), N-acyl-6-OH-sphingosine / ceramide components (for example, 44 as reference values) and N- (α-OH-acyl) -6-OH-sphingosine / ceramide components (for example 43 as reference values) .5), for at least one component selected from the group consisting of 5), the average carbon number of the molecular species contained in the component is calculated as a weighted average value weighted by the abundance ratio of each molecular species, and the carbon number It is possible to determine psoriasis by comparing the weighted average value of with the reference value.

  Further, in the case of psoriasis as the above skin disease, skin quality or skin condition, N-acyl-dihydrosphingosine / ceramide component (reference value, for example, 1% by weight), N-acyl-sphingosine / ceramide component (reference value, for example, 1% by weight), N-acyl-6-OH-sphingosine / ceramide component (reference value, for example, 1% by weight), N- (α-OH-acyl) -sphingosine / ceramide component (reference value, for example, 1% by weight) %), N- (α-OH-acyl) -6-OH-sphingosine / ceramide component (reference value, for example, 1% by weight), N- (α-OH-acyl) -phytosphingosine / ceramide component (reference value) For example, 1% by weight) and N- (ω-OH-acyl) acyl-sphingosine ceramide component (for example, 20 For at least one component selected from the group consisting of%), the percentage unsaturation of 1 or more often species can determine psoriasis by comparing with the reference value.

  Furthermore, in the case of psoriasis as the above skin disease, skin quality or skin condition, the amount per 1 μg of protein of 34 molecular species in the total N-acyl-sphingosine / ceramide component (for example, 0.07 ( ng) and / or psoriasis can be determined by comparing the composition ratio (for example, 3.0% by weight as a reference value) to the total amount of all N-acyl-sphingosine ceramide components with the reference value.

  On the other hand, in the case of atopic dermatitis as the skin disease, skin quality or skin condition, N-acyl-sphingosine / ceramide component (10% by weight as a reference value), N-acyl-6-OH-sphingosine / ceramide component (As a reference value, for example, 19% by weight), N-acyl-phytosphingosine / ceramide component (as a reference value, for example, 15% by weight), N- (α-OH-acyl) sphingosine / ceramide component (as a reference value, for example, 6% by weight), N- (ω-OH-acyl) acyl-6-OH-sphingosine / ceramide component (for example, 1.5% by weight as reference value) and N- (ω-OH-acyl) acyl-phytosphingosine -Ceramide with respect to at least one component selected from the group consisting of ceramide components (for example, 0.5% by weight as a reference value) Determining atopic dermatitis composition ratio to the amount by comparing with the reference value (trends and severity. Hereinafter, the same) can be.

  Further, in the case of atopic dermatitis as the skin disease, skin quality or skin condition, N-acyl-dihydrosphingosine / ceramide component (46.5 as a reference value), N-acyl-sphingosine / ceramide component (standard) As values, for example, 46), N-acyl-6-OH-sphingosine / ceramide component (for example, 44 as reference values) and N- (α-OH-acyl) -6-OH-sphingosine / ceramide component (reference) As a value, for example, for at least one component selected from the group consisting of 44), the average carbon number of the molecular species contained in the component is calculated as a weighted average value with the abundance ratio of each molecular species as a weight, Atopic dermatitis can be determined by comparing the weighted average value of the carbon number.

  Further, in the case of atopic dermatitis as the above skin disease, skin quality or skin condition, N-acyl-6-OH-sphingosine / ceramide component (for example, 1% by weight as a reference value) and / or N- (ω-OH -Regarding the acyl) acyl-sphingosine-ceramide component (as a reference value, for example, 20% by weight), atopic dermatitis can be determined by comparing the ratio of molecular species having one or more unsaturated bonds to the reference value. it can.

  Furthermore, in the case of atopic dermatitis as the above skin disease, skin quality or skin condition, the amount per 1 μg of protein of 34 molecular species in all N-acyl-sphingosine / ceramide components (for example, 0 as a reference value) .04 (ng)) and / or the composition ratio of the total N-acyl-sphingosine / ceramide component to the total amount (for example, 2.5% by weight as a reference value) is compared with the reference value to determine atopic dermatitis can do.

  ADVANTAGE OF THE INVENTION According to this invention, the skin quality of a test subject can be objectively evaluated and the skin quality determination method which can be utilized for diagnosis of various skin related diseases, skin quality, or a skin state can be provided. According to the skin quality determination method of the present invention, for example, it is possible to monitor changes in skin quality when a substance that affects the skin, such as an external preparation for skin, cosmetics, or quasi drugs, is administered. The presence or absence of an effect on the subject can also be determined.

Hereinafter, the present invention will be described in detail with reference to the drawings.
The present invention is a skin quality determination method capable of objectively evaluating the skin quality of a subject. The skin quality determination method according to the present invention is based on lipid composition information obtained from a subject, information on lipid components whose composition ratio changes in relation to skin quality, lipid composition information obtained from a healthy individual, and specific skin quality The skin quality of the subject is determined by comparison with a reference value calculated based on the lipid composition information obtained from the individual. Examples of the subject include animals such as humans, monkeys, chimpanzees, dogs, cats, cows, pigs, rats, and mice. When an animal as described above is applied as a subject, lipid composition information can be obtained from the skin stratum corneum extract of the animal. In addition, as the subject, the above-mentioned humans and animals, skin and cadaver skin collected from living bodies, cells, reconstructed cells and skin tissues, stratum corneum, blood, body fluid, hair, scalp and organs are used. You can also. In particular, as a subject, it is preferable to use a skin stratum corneum collected from a site where the skin quality is to be evaluated or from the vicinity thereof. When the skin stratum corneum is collected from a predetermined site, there is no particular limitation. For example, a method of applying an adhesive surface of an adhesive tape to the site and then peeling the tape can be applied. At this time, it is preferable to perform a pretreatment such as removing body hair, hair, sebum components present on the surface, and impurities on the site where the skin stratum corneum is collected.

  In the present invention, the lipid composition information means information indicating the abundance or abundance ratio of various lipid components extracted from the skin stratum corneum. Examples of the lipid component include components derived from organisms having a long-chain fatty acid or a hydrocarbon chain, and include fatty acids, glycerides, wax esters, sphingolipids, phospholipids, cholesterol, and the like. In particular, in the present invention, it is preferable to obtain lipid composition information indicating the type of ceramide (ceramide class), which is a kind of sphingolipid, and the composition ratio of the molecular species constituting each ceramide class. Here, ceramide means a compound having a structure in which a fatty acid is bonded with an acid amide to the amino group of a sphingosine base. Therefore, ceramides include various ceramide classes depending on the combination of sphingosine base and fatty acid.

  Examples of the ceramide class include N- (ω-OH-acyl) acyl-dihydrosphingosine (EODS), N- (ω-OH-acyl) acyl-sphingosine (EOS), and N- (ω-OH-acyl) acyl. -Phytophingosine (EOP), N- (ω-OH-acyl) acyl-6-OH-sphingosine (EOH), N-acyl-dihydrosphingosine (NDS), N-acyl-sphingosine (NS), N-acyl-phystophosphines (NOH) NP), N-acyl-6-OH-sphingosine (NH), N- (α-OH-acyl) -dihydrosphingosine (ADS), N- (α-OH-acyl) -sphingosine (AS), N- (α-OH-acyl) -phytosphingosine (AP) and N- (α-OH-acyl) can be exemplified -6-OH-sphingosine (AH).

  These ceramide classes include N- (ω-OH-acyl) acyl-dihydrosphingosine ceramide (sometimes referred to as Cer [EODS]), N- (ω-OH-acyl) acyl-sphingosine ceramide (Cer [ EOS], N- (ω-OH-acyl) acyl-phytosphingosine ceramide (sometimes referred to as Cer [EOP]), N- (ω-OH-acyl) acyl-6-OH -Sphingosine ceramide (sometimes referred to as Cer [EOH]), N-acyl-dihydrosphingosine ceramide (sometimes referred to as Cer [NDS]), N-acyl-sphingosine ceramide (referred to as Cer [NS]) N-acyl-phytosphingosine ceramide (sometimes referred to as Cer [NP]), N-acyl-6-OH-sphingosine ceramide (also referred to as Cer [NH]). N- (α-OH-acyl) -dihydrosphingosine ceramide (sometimes referred to as Cer [ADS]), N- (α-OH-acyl) -sphingosine ceramide (Cer [AS]) N- (α-OH-acyl) -phytosphingosine ceramide (sometimes referred to as Cer [AP]) and N- (α-OH-acyl) -6-OH-sphingosine ceramide (Cer [ AH] may also be referred to). The chemical formulas of these 12 kinds of ceramides are described below.

  These ceramide classes include various molecular species depending on the carbon number of fatty acids and sphingoids, the degree of unsaturation of fatty acids and sphingoids, and / or the position of unsaturated bonds contained in fatty acids and sphingoids. It will be. That is, in the present invention, the lipid composition information is preferably information indicating the abundance or abundance ratio of molecular species included in each class for each ceramide class.

  As a method for acquiring the lipid composition information, for example, a conventionally known thin layer chromatography method using a silica gel plate can be applied. According to the lipid analysis method using thin layer chromatography (TLC), it is possible to analyze lipids by class or type, but it may be difficult to obtain information by lipid molecule. Therefore, as a method of overcoming the drawbacks of this thin layer chromatographic method, lipids are separated by normal phase liquid chromatography, ionized by electrospray ionization (ESI), and detected by a mass spectrometer. Lipid composition information may be obtained by applying a method for identifying the chemical structure of a molecule (see JP 2003-28849 A).

In addition, methods for obtaining lipid composition information include gas chromatograph mass spectrometer (GC-MS), TLC, liquid chromatograph mass spectrometer (LC-MS or LC-MS / MS), liquid chromatograph time-of-flight type Mass spectrometer (LC-TOF-MS or LC-TOF-MS / MS), matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS or MALDI-TOF-MS / MS), nuclear magnetic resonance (NMR) apparatus etc. can be used individually or in combination. These methods are described in the following literature: Raith K, Darius J, Neubert RH. Ceramide analysis utilizing gas chromatography-mass spectrometry. J Chromatogr A. 2000 Apr 21; 876 (1-2): 229-33; Nakamura K , Suzuki Y, Goto-Inoue N, Yoshida-Noro C, Suzuki A. Structural characterization of neutral glycosphingolipids by thin-layer chromatography coupled to matrix-assisted laser desorption / ionization quadrupoleion trap time-of-flight MS / MS. 2006 Aug 15; 78 (16): 5736-43; Farwanah H, Wohlrab J, Neubert RH, Raith K. Profiling of human stratum corneum ceramides by means of normal phase LC / APCI-MS. Anal Bioanal Chem. 2005 Oct; 383 (4): 632-7. Epub 2005 Oct 19; and Ponec M, Weerheim A, Lankhorst P, Wertz P. New acylceramide in native and reconstructed epidermis. J Invest Dermatol. 2003 Apr; 120 (4): 581-8 You can refer to it.
Furthermore, as a technique for acquiring lipid composition information, it is particularly preferable to apply the technique disclosed in Japanese Patent Application No. 2005-300251. In summary, this method separates a lipid sample prepared from a sample collected from the stratum corneum with a liquid chromatograph, ionizes and detects it with a mass spectrometer, and displays the data output from the mass spectrometer with retention time and mass. This is a method of analysis by a multistage mass chromatogram developed on three axes of number / charge (m / z) and ionic strength.

  Specifically, for example, lipid composition information can be acquired using an analysis system 1 as shown in FIG. Hereinafter, a method of analyzing ceramide molecular species using the analysis system 1 shown in FIG. 1 will be described. The analysis system 1 includes a liquid chromatograph 10, an ionization promoting liquid delivery device 20, a mass spectrometer 30, and a calculation device 40.

  The liquid chromatograph 10 includes high-pressure gradient pumps 11a and 11b for feeding two kinds of eluents a and b, an autoinjector 12 for introducing a lipid sample solution d, a guard column 13 and a separation column 14. Here, as the lipid sample solution d, a sample solution containing ceramide prepared by extraction or the like from a sample of the skin stratum corneum, or skin, cadaver skin or stratum corneum, cells, reconstructed from a human, an animal, or a living body A sample solution containing ceramide prepared by extraction or the like from collected cells, skin tissue, blood, body fluid, hair, scalp, organ, or the like is used.

  As a method for collecting the skin stratum corneum, a method with low risk to the subject and low burden is preferable. Examples of such a sampling method include a tape peeling method (tape stripping method). As a method of preparing a lipid sample solution d containing ceramide from a sample of skin stratum corneum peeled off with tape, ceramide is extracted from the sample using a solvent in which the solubility of ceramide is high and the tape is difficult to dissolve. It is preferable to do. Examples of such a solvent include methanol, ethanol, isopropanol and the like.

  Moreover, as a method for preparing the lipid sample solution d from the sample, a known method can be applied as a method for separating and purifying a lipid component from a biological sample. As an example, the lipid sample solution d can be prepared from the culture using the Bligh and Dyer method. In addition, the lipid sample solution d can be prepared from the culture by applying the Folch method or the like.

  On the other hand, as the eluents a and b, those that are mixed well, can be separated by ceramide and class while appropriately holding ceramide, and do not contain non-volatile acids and salts at high concentrations. preferable. As a combination example of the eluents a and b, for example, the eluent a can be a mixed liquid of hexane / isopropanol and the eluent b can be a mixed liquid of hexane / isopropanol / water.

The guard column 13 is provided as necessary to protect the separation column 14 and is filled with the same filler as the separation column 14. As a packing material for the guard column 13 and the separation column 14, for example, silica gel or a highly polar column such as silica gel bonded with a diol group, a CN group, or an NH ₂ group can be used.

  By configuring the liquid chromatograph 10 as described above, the lipid containing ceramide can be separated based on the difference in polarity. The lipid containing ceramide separated by the liquid chromatograph 10 will be introduced into the ionization promoting liquid delivery device 20 at the subsequent stage.

  The ionization promoting liquid delivery apparatus 20 includes a pump 21 for delivering the ionization promoting liquid c, and a tee connector 22 for mixing the eluent from the separation column 14 and the ionization promoting liquid c. The ionization promoting liquid c is mixed with the eluent from the separation column 14 via the tee connector 22. The mixed solution is introduced into the ionization device 31 of the mass spectrometer 30.

  The ionization accelerating liquid c is used to improve that it is difficult to obtain sufficient ionization efficiency with ESI when a low polarity solvent such as hexane is used as an eluent. As the ionization accelerating liquid c, a liquid having a property such as surface tension, viscosity, ion generation ability, and solvating power suitable for ionizing the eluent by mixing well with the eluent is appropriately selected. For example, when hexane is used as an eluent, a polar solvent such as isopropanol, ethanol, or methanol is used as the ionization promoting liquid c.

In the ionization promoting liquid c, [M + H] ⁺ and [M + H−H ₂ O] ⁺ are detected with high sensitivity in the positive ion mode, and [M−H] ⁻ and [M + HCOO] ⁻ are detected with high sensitivity in the negative ion mode. As detected, salts such as ammonium formate, ammonium acetate, etc. can be added. In addition, a volatile acid such as formic acid, acetic acid, and trifluoroacetic acid may be added to the ionization promoting liquid c.

  The mass spectrometer 30 includes an ionizer 31 and a mass separation detector 32. A mixed solution of an ionization promoting solution and an eluent is introduced through the tee connector 22 to ionize lipid components including ceramide. .

  Electrospray ionization (ESI) is preferred as an ionization method in the ionizer 31 from the viewpoint of sensitivity, but atmospheric pressure chemical ionization (APCI), atmospheric pressure photoionization (APPI), and the like are also used. You can also.

  The mass separation detection device 32 separates and detects ions generated by the ionization device 31 for each m / z. As the mass separation detector 32, a mass spectrometer such as a quadrupole (Q) mass spectrometer or an ion trap (IT) mass spectrometer, or a hybrid mass spectrometer such as a Q-TOF type or IT-TOF type is used. Alternatively, a tandem mass spectrometer (MS / MS) such as a triple quadrupole type can be used.

  The computing device 40 has computing means that develops the retention time in the liquid chromatograph 10, the m / z and the ion intensity detected by the mass spectrometer 30 on three axes, and forms a multistage mass chromatogram. Further, although not shown, it is preferable that the arithmetic unit 40 be able to access a database in which the retention time and m / z are associated with each molecular species belonging to each ceramide class. Further, the computing device 40 uses the multistage mass chromatogram formed by the computing means as input data, searches the database based on the peak retention time and m / z included in the input multistage mass chromatogram, It is preferable to have a comparison calculation means for specifying the ceramide molecular species corresponding to each peak. Furthermore, the calculation device 40 has a display means for outputting and displaying the multistage mass chromatogram formed by the calculation means and / or the ceramide molecular species corresponding to each peak specified by the comparison calculation means in a desired format. Is preferred.

  In the analysis system configured as described above, as the liquid chromatograph 10 and the mass spectrometer 30, a commercially available liquid chromatograph-mass spectrometer in which these are integrated can be used.

  In the analysis system shown in FIG. 1, a multistage mass chromatogram as shown in FIG. 2 is shown as an example in which all data detected by the mass spectrometer 30 are developed on three axes of retention time, m / z, and ion intensity. Can be output. According to the multistage mass chromatogram, all the ceramide molecules contained in the lipid sample solution can be captured comprehensively, and the recognition of homologues and trace components becomes easy, so from the information on retention time and m / z Individual ceramide molecules can be easily identified. For example, ceramides such as ceramide EODS, EOS, EOP, EOH, NDS, NS, NP, NH, ADS, AS, AP and AH can be separated and analyzed, and the total carbon in each ceramide class It is possible to separate and analyze molecular species having different numbers, degrees of unsaturation of fatty acids and sphingoids, and positions of unsaturated bonds contained in fatty acids and sphingoids.

  In the present analysis system 1, the computing device 40 accesses a database in which retention times and m / z are associated with each ceramide molecular species belonging to each ceramide class, and is included in the output multistage mass chromatogram. Each peak can be identified based on the retention time of each peak and m / z. That is, when the computing device 40 outputs a multistage mass chromatogram, information on the molecular species of each peak (for example, the total number of carbon atoms, the degree of unsaturation of fatty acids and sphingoids, and unsaturated bonds contained in fatty acids and sphingoids). Can be linked and output.

  In the present analysis system 1, a plurality of specific ceramide molecules can be detected with high sensitivity by performing a selected ion monitoring method in the mass spectrometer 30. Also for the detection data by the selected ion monitoring method, a multistage mass chromatogram similar to that in FIG. 2 can be formed, and a plurality of specific ceramide molecules can be comprehensively captured.

FIG. 2 is an example of a multistage mass chromatogram in which a lipid sample actually prepared from a sample of the stratum corneum is analyzed by the analysis system 1. As shown in the figure, according to this multi-stage mass chromatogram, the lipid sample solution d contains 12 classes of ceramide, and it is also possible to discriminate homologues having different carbon numbers and unsaturation levels of these ceramides. It can be seen that various ceramide molecules contained in the lipid sample solution d can be comprehensively identified. Although the figure shows only the measurement result in the positive ion mode, the ion species observed differs depending on the type of ceramide, and [M + H] ⁺ is easily observed, and [M + H−H ₂ O] ⁺ is Since some of them are easily observed, it is preferable to analyze the measurement results in the negative ion mode as well.

  In the analysis of ceramide, it is possible to handle quantitatively only by comparing the peak areas of each ceramide molecule. However, in order to perform quantitative handling more accurately, an internal standard substance is added to the lipid sample solution d. It is preferable to determine the peak area ratio of each ceramide molecule to the peak area of the internal standard substance. More preferably, a standard product of all ceramide molecules is prepared and the absolute amount is calculated from the absolute calibration curve. When using an internal standard substance, it is preferable to use a ceramide molecule that is hardly present in the skin stratum corneum. For example, N-heptadecanyl-D-erythro-sphingosine ceramide is used.

  When determining the absolute or relative amount of each ceramide molecule, measure the area of the exfoliated stratum corneum, the weight of the stratum corneum, the amount of protein, the number of cells, etc., and divide by one of these values. Therefore, it is preferable to standardize.

  In the present invention, as described above, the multistage mass chromatogram output by the analysis system 1 can be used as the lipid composition information obtained from the subject. In the skin quality determination method according to the present invention, lipid composition information obtained from a subject is compared with a reference value. Here, the reference value is a specific value defined for a predetermined lipid composition relating to a specific skin quality, and is obtained from a lipid composition information obtained from a healthy individual and a lipid composition information obtained from an individual of a specific skin quality. Calculated by comparison. Here, the specific skin quality can include the skin quality of patients with skin-related diseases such as the skin quality of patients with psoriasis and the skin quality of patients with atopic dermatitis. Specific skin types include sensitive skin, dry skin, oily skin, skin with poor moisturizing power, skin with poor barrier function, skin that is prone to acne, skin that is prone to scaling, and skin that is prone to erythema It means to include the condition of the skin including the scalp.

  Moreover, when calculating a reference value, although it does not specifically limit, lipid composition information is first acquired about a healthy subject group and a specific skin quality person group as mentioned above. Next, the lipid composition information obtained from the healthy group is compared with the lipid composition information obtained from the specific skin group. At this time, when a multistage mass chromatogram as described above is acquired as the lipid composition information, the healthy group and the specific skin quality group may be compared for a specific molecular species in the ceramide class. Such comparisons identify lipid components (eg, ceramide class molecular species) that are significantly different in abundance, composition ratio, etc. between the healthy group and the specific skin type group, It is possible to calculate the abundance and composition ratio of the lipid component characteristic to the skin quality person. The value calculated in this way can be defined as the reference value.

<Example of reference value 1>
Specifically, in the psoriasis patient group, the composition ratio of Cer [NS], Cer [NP], and Cer [AS] to the total amount of ceramide in the ceramide class is significantly different from the healthy group. . Specifically, Cer [NS] in the psoriasis patient group has a significantly increased composition ratio relative to the total amount of ceramide compared to the healthy group. Moreover, Cer [NP] in the psoriasis patient group has a significant decrease in the composition ratio with respect to the total amount of ceramide compared to the healthy group. Furthermore, Cer [AS] in the psoriasis patient group has a significant increase in the composition ratio relative to the total amount of ceramide compared to the healthy group.

  In the group of patients with atopic dermatitis, ceramides of Cer [NS], Cer [NH], Cer [NP], Cer [AS], Cer [EOH] and Cer [EOP] are among the ceramide classes. The composition ratio with respect to the total amount is significantly different from that of the healthy subject group. Specifically, Cer [NS] in the atopic dermatitis patient group has a significant increase in the composition ratio relative to the total amount of ceramide compared to the healthy group. Moreover, Cer [NH] in the atopic dermatitis patient group has a significant decrease in the composition ratio with respect to the total amount of ceramide compared with the healthy subject group. Furthermore, Cer [NP] in the atopic dermatitis patient group has a significantly reduced composition ratio with respect to the total amount of ceramide compared to the healthy group. Furthermore, Cer [AS] in the atopic dermatitis patient group has a significant increase in the composition ratio relative to the total amount of ceramide compared to the healthy group. Furthermore, Cer [EOH] in the atopic dermatitis patient group has a significant decrease in the composition ratio relative to the total amount of ceramide compared to the healthy group. Furthermore, Cer [EOP] in the atopic dermatitis patient group has a significantly reduced composition ratio relative to the total amount of ceramide compared to the healthy group.

  Although details will be described later, specifically, as shown in Table 1, for example, a reference value can be defined, and how the reference value is determined to be an abnormal value can be defined. In Table 1 below, for example, the NS column in the row of atopic dermatitis is described as “10 or more” because the composition ratio of Cer [NS] to the total amount of Ceramide in the lipid composition information collected from the subject Means that it is recognized as an abnormal value when it is 10% by weight or more.

  An abnormal value in the lipid composition information collected from the subject can be detected by comparing the reference value regarding the composition ratio of the ceramide class shown in Table 1 with the lipid composition information collected from the subject. Then, by referring to the number and degree of ceramide classes showing abnormal values, the skin quality of the subject is similar to that of psoriasis patients or similar to that of atopic dermatitis patients, or the skin of healthy subjects It can be judged whether it is quality.

  In particular, in the lipid composition information collected from the subject, it can be judged that there is a tendency for atopic dermatitis if there are 1 or more abnormal values exceeding the reference values shown in Table 1, and there are 3 or more abnormal values. Then, it can be judged that atopic dermatitis is serious. In addition, in the lipid composition information collected from the subject, if the abnormal value exceeding the reference value shown in Table 1 is one or more of NS, NP and AS, it can be determined that there is a tendency for psoriasis. If all three AS values are abnormal, it can be determined that psoriasis is severe.

<Example 2 of reference value>
In the psoriasis patient group, the average carbon number of the molecular species included in Cer [NDS], Cer [NS], Cer [NH] and Cer [AH] is significantly different from that in the healthy group among the ceramide class. ing. Here, the average carbon number means a weighted average value that is calculated by using the abundance ratio of each molecular species as a weight for a plurality of molecular species having different carbon numbers. Specifically, the mean carbon numbers of Cer [NDS], Cer [NS], Cer [NH] and Cer [AH] in the psoriasis patient group are all significantly reduced compared to the healthy group.

  In the atopic dermatitis patient group, the average carbon number of molecular species included in Cer [NDS], Cer [NS], Cer [NH] and Cer [AH] is particularly healthy among the ceramide class. Significantly different from the group. Here, as described above, the average carbon number means a weighted average value that is calculated by using the abundance ratio of each molecular species as a weight for a plurality of types of molecular species having different carbon numbers. Specifically, the average carbon number of Cer [NDS], Cer [NS], Cer [NH] and Cer [AH] in all patients with atopic dermatitis is significantly reduced compared to the healthy group. Yes.

  Although details will be described later, specifically, for example, a reference value and an abnormal value can be defined as shown in Table 2. In Table 2 below, for example, the NS column in the row of atopic dermatitis is described as “less than 46” because the average carbon of Cer [NS] molecular species from the lipid composition information collected from the subject This means that the number is calculated, and if this value is less than 46, it is recognized as an abnormal value.

  Detecting abnormal values in the lipid composition information collected from the subject by comparing the reference value regarding the average carbon number of the molecular species included in the ceramide class shown in Table 2 with the lipid composition information collected from the subject. Can do. Then, by referring to the number and degree of ceramide classes showing abnormal values, the skin quality of the subject is similar to that of psoriasis patients or similar to that of atopic dermatitis patients, or the skin of healthy subjects It can be judged whether it is quality.

  In particular, in the lipid composition information collected from the subject, if either NDS or NS shown in Table 2 is an abnormal value, it can be determined that there is a tendency for atopic dermatitis. If it is an abnormal value, it can be judged that atopic dermatitis is serious. In addition, in the lipid composition information collected from the subject, if either NDS or NS shown in Table 2 is abnormal, it can be determined that there is a tendency for psoriasis. In addition, a total of three or more abnormal values are abnormal. If so, it can be judged that psoriasis is severe.

<Example 3 of reference value>
In the psoriasis patient group, the composition ratio of the molecular species having 34 carbon atoms in Cer [NS] is significantly increased in the ceramide class. In the atopic dermatitis patient group, the composition ratio of molecular species having 34 carbon atoms in Cer [NS] is particularly significantly increased in the ceramide class.

  Although details will be described later, specifically, for example, as shown in Table 3, a reference value and an abnormal value can be defined. In Table 3 below, for example, the column of the molecular weight of 34 carbon atoms in Cer [NS] in the row of atopic dermatitis is described as “0.04 or more and less than 0.07”, which was collected from subjects. This means that the molecular species having 34 carbon atoms in Cer [NS] is quantified from the lipid composition information, and when this value is 0.04 or more and less than 0.07, it is recognized as an abnormal value.

  As shown in Table 3, the reference value regarding the amount and / or composition ratio of the molecular species having 34 carbon atoms in Cer [NS] was collected from the subject by comparing it with the lipid composition information collected from the subject. An abnormal value in the lipid composition information can be detected. Then, by referring to the number and degree of ceramide classes showing abnormal values, the skin quality of the subject is similar to that of psoriasis patients or similar to that of atopic dermatitis patients, or the skin of healthy subjects It can be judged whether it is quality.

  In particular, in the lipid composition information collected from the subject, when one of the molecular species amount and the composition ratio shown in Table 3 is an abnormal value, it can be determined that there is a tendency for atopic dermatitis, and both are abnormal values. If it is, it can be judged that atopic dermatitis is severe. In addition, in the lipid composition information collected from the subject, when one of the molecular species amount and the composition ratio shown in Table 3 is an abnormal value, it can be determined that there is a tendency for psoriasis, and both are abnormal values It can be judged that psoriasis is severe.

<Example 4 of reference value>
In the psoriasis patient group, among Ceramide class, Cer [NDS], Cer [NS], Cer [NH], Cer [AS], Cer [AH], Cer [AP] and Cer [EOS] Molecular species with one or more unsaturated bonds are significantly increased.

  In the atopic dermatitis patient group, among the ceramide class, especially for Cer [NH] or Cer [EOS], molecular species having one or more unsaturated bonds are significantly increased. Therefore, due to the test substance, the molecular species having one or more unsaturated bonds are significantly reduced in Cer [NH] and Cer [EOS] in the subject.

  Although details will be described later, specifically, for example, as shown in Table 4, a reference value and an abnormal value can be defined. In Table 4 below, for example, Cer [NS] in the line of psoriasis: the column of NS amount / NS total amount of 2 or more unsaturateds is described as “1 or more” because the lipid composition collected from the subject Calculate the ratio of two or more unsaturated molecular species in Cer [NS] from the information, and if this value is 1% by weight or more with respect to the total amount of Cer [NS] I mean.

  As shown in Table 4, samples were taken from subjects by comparing reference values for the proportion of molecular species with one or more unsaturated bonds in a particular ceramide class with lipid composition information collected from subjects. An abnormal value in the lipid composition information can be detected. Then, by referring to the number and degree of ceramide classes showing abnormal values, the skin quality of the subject is similar to that of psoriasis patients or similar to that of atopic dermatitis patients, or the skin of healthy subjects It can be judged whether it is quality.

  In particular, in the lipid composition information collected from the subject, when one of the molecular species having one or more unsaturated bonds in NH and EOS shown in Table 4 has an abnormal value, there is a tendency for atopic dermatitis. If both are abnormal values, it can be determined that atopic dermatitis is severe. In addition, in the lipid composition information collected from the subject, if any one of NDS, NS, AS, AH and AP shown in Table 4 is abnormal, it is judged that there is a tendency for psoriasis, and NDS, NS, NH, AS Psoriasis can be judged to be severe if it is an abnormal value of 3 or more of AH, AP and EOS.

  EXAMPLES Hereinafter, although this invention is demonstrated in detail using an Example, the technical scope of this invention is not limited to a following example.

[Example 1]
In this example, a ceramide component, which is a kind of lipid component, was analyzed using a skin stratum corneum collected from a subject.

Atopic dermatitis patient 8 people attending to the subject dermatology clinic (16 to 36 years) and healthy volunteers seven corresponding to it (25 to 37 years old), psoriasis patients 10 people attending the dermatology clinic (36 to 74 years old ) And 9 healthy volunteers (39-76 years old), 7 dry skins, 6 normal skins, 12 skins with low barrier function in their 30s (female), and 10 normal skins in their 30s (female) )

Extraction of skin stratum corneum In dry skin test subjects, normal skin test subjects forearm flexion side, in any patient the skin rash part of the arm and the adjacent non-rash part, in healthy subjects press the tape on the same site as the patient in the arm, The skin stratum corneum was peeled off. In females with low barrier function and normal skin, tape was pressed against the same part of the cheek to peel the skin stratum corneum. Each tape was cut in half, one was used for ceramide component analysis and the other was used for protein quantification. For protein quantification, add 0.1N NaOH, 1% SDS aqueous solution to the other half of the tape, heat at 60 ° C for 2 hours, solubilize the protein, cool to room temperature, and then add 2N HCl to neutralize, Using BCA Protein Assay, the quantitative value of the protein was obtained from the calibration curve by BSA.

Extraction of Lipid Molecules Methanol containing N-heptadecanyl-D-erythro-sphingosine was added to the tape from which the skin stratum corneum was collected, and lipid molecules were extracted by irradiating with ultrasonic waves.

Preparation of crude fraction of ceramide and sample solution The above methanol extract was dried under a nitrogen stream and dissolved by adding chloroform / methanol = 99.5 / 0.5 (v / v) to a silica gel cartridge for solid phase extraction. Applied. After sufficiently applying chloroform / methanol = 99.5 / 0.5 (v / v), chloroform / methanol = 95/5 (v / v) was applied to obtain the eluate. The eluate was dried under a nitrogen stream and then dissolved by adding hexane / isopropanol / formic acid = 95/5 / 0.1 (v / v / v) to obtain a sample solution.

Analysis conditions of ceramide component In this example, a liquid chromatograph-mass spectrometer was used. Specifically, Yokogawa Analytical Systems, Inc., Agilent 1100 Series LC / MSD was used as an analysis system in which a liquid chromatograph and a mass spectrometer were integrated.
As a separation column, GL Sciences Inc., Inertsil SIL 100A-3, 1.5 mmφ × 150 mm (3 μm) was used. As a guard column, GL Sciences Inc., Inertsil SIL 100A-3, 1.5 mmφ × 10 mm (3 μm) was used. As eluent a (see FIG. 1), hexane / isopropanol / formic acid = 95/5 / 0.1 (v / v / v) is used, and as eluent b (see FIG. 1), hexane / isopropanol / 50 mmol / L. Ammonium formate aqueous solution = 25/65/10 (v / v / v) was used. The gradient conditions are shown in Table 5.

  As the ionization promoting solution, isopropanol / 5 mmol / L ammonium formate methanol solution = 50/50 (v / v) was used. The flow rate of the ionization promoting solution was 0.1 mL / min.

  The analysis conditions in the mass spectrometer are as follows. Ionization method: ESI, polarity: positive ion, measurement mass range: 250-1500, fragmentor voltage: 150V, Vcap voltage: 3500V, nebulizer pressure: 20psig, drying gas temperature: 300 ° C, drying gas flow rate: 8L / min

Results The data obtained from the mass spectrometer was developed into a multistage mass chromatogram having three axes of retention time, m / z and ionic strength. Thereafter, each peak included in the multistage mass chromatogram was identified using a database storing information on retention time and m / z for each known ceramide molecular species. Then, the peak area of each ceramide molecule was determined, the peak area ratio with respect to the internal standard substance was calculated, and further divided by the protein amount, thereby calculating the relative amount of each ceramide molecule per unit protein amount. The absolute amount of each ceramide molecular species per unit protein amount was calculated by multiplying these by the detection sensitivity correction coefficient for each ceramide molecular species determined in advance.

  The result of comparing the composition ratio of the ceramide component in the atopic dermatitis patient group and the healthy group is shown in FIG. 3, and the result of comparing the composition ratio of the ceramide component in the psoriasis patient group and the healthy group is shown in FIG. From the results of FIGS. 3 and 4, it has been clarified that the class composition of ceramide is greatly different between the atopic dermatitis patient and the psoriasis patient as compared with the healthy subject.

  In particular, as shown in FIG. 3, the composition ratio of Cer [NS] in the atopic dermatitis patient group to the total amount of ceramide is significantly higher than that in the healthy group, and the Cer [ The composition ratio of NH] to the total amount of ceramide was significantly decreased compared to the healthy group, and the composition ratio of Cer [NP] to the total amount of ceramide of the patient with atopic dermatitis was significantly compared to the healthy group. The composition ratio of Cer [AS] to the total amount of ceramide in the atopic dermatitis patient group was significantly higher than that in the healthy group, and the Cer [EOH] ceramide in the atopic dermatitis patient group was significantly increased. The composition ratio to the total amount was significantly reduced compared to the healthy group, and the composition ratio of Cer [EOP] in the atopic dermatitis patient group to the total amount of ceramide was significantly decreased compared to the healthy group. Yes.

  In particular, as shown in FIG. 4, the composition ratio of Cer [NS] in the psoriasis patient group to the total amount of ceramide was significantly higher than that in the healthy group, and the composition of Cer [NP] in the patient group of psoriasis to the total amount of ceramide. The ratio was significantly decreased compared to the healthy group, and the composition ratio of Cer [AS] to the total amount of ceramide in the psoriasis patient group was significantly increased compared to the healthy group.

  Of the characteristics shown in FIGS. 3 and 4, the composition ratio of Cer [NP] to the total amount of ceramide clearly shows a very low value in patients with atopic dermatitis and psoriasis compared with the healthy group. became.

  From the results shown in FIGS. 3 and 4, for the ceramide class in which the composition ratio is significantly different between the psoriasis patient group and the atopic dermatitis patient group and the healthy subject group, the reference value is set as shown in Table 1 above. Can be calculated.

  On the other hand, FIG. 5 shows the results of comparing the weighted average values with weights of the abundance ratios of the carbon species of the molecular species constituting the ceramide class in the atopic dermatitis patient group and the healthy person group. FIG. 6 shows the result of comparing the weighted average values with the abundance ratios as weights for the carbon numbers of the molecular species constituting the ceramide class in the group. From the results of FIGS. 5 and 6, it became clear that the weighted average value of the total carbon number tends to be smaller in atopic dermatitis and psoriasis than in the healthy subject group. In particular, as shown in FIG. 5, the average carbon number of molecular species contained in Cer [NDS], Cer [NS], Cer [NH] and Cer [AH] in the atopic dermatitis patient group is significantly different from that in the healthy subject group. Has decreased. In addition, as shown in FIG. 6, the molecular species contained in Cer [NDS], Cer [NS], Cer [NH], Cer [ADS], Cer [AS] and Cer [AH] of the atopic dermatitis patient group The average carbon number is significantly reduced compared with the healthy group.

  From the results shown in FIGS. 5 and 6, the ceramide class in which the average carbon number of the molecular species is significantly different between the psoriasis patient group and the atopic dermatitis patient group and the healthy subject group is shown in Table 2 above. A reference value can be calculated as shown.

  Further, FIG. 7 shows the result of comparing the amount per 34 μg of protein of the molecular species having 34 carbon atoms among Cer [NS] in the atopic dermatitis patient group and the healthy subject group. Cer in the psoriasis patient group and the healthy subject group is shown in FIG. FIG. 8 shows the result of comparing the amount per [μg] of the protein of 34 molecular species in [NS]. Moreover, the result of comparing the composition ratio of the molecular species having 34 carbon atoms to the total amount of Cer [NS] in Cer [NS] in the atopic dermatitis patient group and the healthy subject group is shown in FIG. FIG. 10 shows the result of comparing the composition ratio of the molecular species having 34 carbon atoms to the total amount of Cer [NS] among the Cer [NS] in the group.

  As shown in FIGS. 7 to 10, in atopic dermatitis patients and psoriasis patients, the absolute amount of molecular species having a total carbon number of 34 among Cer [NS] and the composition ratio to the total amount of Cer [NS] are healthy subjects. It became clear that there was more compared with the group.

  From the results shown in FIGS. 7 to 10, the amount and composition ratio of the molecular species having 34 carbon atoms in Cer [NS] are significantly different among the psoriasis patient group, the atopic dermatitis patient group, and the healthy subject group. Therefore, the reference value can be calculated as shown in Table 3 above.

  Furthermore, FIG. 11 shows the result of comparing the composition ratio of molecular species having one or more unsaturated bonds for the ceramide class in the atopic dermatitis patient group and the healthy subject group. FIG. 12 shows the result of comparing the composition ratios of molecular species having one or more unsaturated bonds for each class. As shown in FIG. 11, in the atopic dermatitis patient group, especially for Cer [NH] and Cer [EOS], the molecular species having one or more unsaturated bonds are significantly increased. Moreover, as shown in FIG. 12, in the psoriasis patient group, Cer [NDS], Cer [NS], Cer [NH], Cer [AS], Cer [AH], Cer [AP] and Cer [EOS] The molecular species having one or more unsaturated bonds is significantly increased.

  From the results shown in FIGS. 11 and 12, a specific ceramide class in which the composition ratio of the molecular species having one or more unsaturated bonds is different between the psoriasis patient group and the atopic dermatitis patient group and the healthy subject group. As shown in Table 4, the reference value can be calculated.

  The result of comparing the composition ratio of the ceramide component in the dry skin group and the normal skin group is shown in FIG. 13, and the result of comparing the composition ratio of the ceramide component in the skin group with low barrier function in the 30s and the normal skin group in the 30s. As shown in FIG. From the results of FIGS. 13 and 14, it was clarified that the class composition of ceramide is significantly different between the dry skin test subject and the skin test subject with low barrier function as compared with the normal skin.

  In particular, as shown in FIG. 13, in the dry skin group, Cer [NDS], Cer [NS], Cer [NH], Cer [NP], Cer [AS], Cer [ The composition ratio of AH], Cer [AP], Cer [EOS] and Cer [EOH] to the total amount of ceramide is different from that of the normal skin group. Specifically, Cer [NDS] in the dry skin group has a reduced composition ratio to the total amount of ceramide compared to the normal skin group. Cer [NS] has an increased composition ratio to the total amount of ceramide compared to the normal skin group. Further, Cer [NH] in the dry skin group has a reduced composition ratio with respect to the total amount of ceramide as compared with the normal skin group. Furthermore, Cer [NP] in the dry skin group has a reduced composition ratio with respect to the total amount of ceramide compared to the normal skin group. Furthermore, Cer [AS] in the dry skin group has an increased composition ratio relative to the total amount of ceramide compared to the normal skin group. Furthermore, Cer [AH] in the dry skin group has a reduced composition ratio relative to the total amount of ceramide compared to the normal skin group. Furthermore, Cer [AP] in the dry skin group has a reduced composition ratio to the total amount of ceramide compared to the normal skin group. Furthermore, Cer [EOH] in the dry skin group has a reduced composition ratio with respect to the total amount of ceramide compared to the normal skin group.

  In the skin group with low barrier function, as shown in FIG. 14, among the ceramide classes, in particular, Cer [NDS], Cer [NS], Cer [NH], Cer [NP], Cer [AS] The composition ratio of Cer [EOS] and Cer [EOH] to the total amount of ceramide is significantly different from that of the normal skin group. Specifically, Cer [NDS] in the skin group with a low barrier function has a significantly increased composition ratio with respect to the total amount of ceramide compared to the normal skin group. In addition, Cer [NS] in the skin group with a low barrier function has a significantly increased composition ratio with respect to the total amount of ceramide as compared with the normal skin group. Furthermore, Cer [NH] in the skin group with a low barrier function has a significantly reduced composition ratio with respect to the total amount of ceramide compared with the normal skin group. Furthermore, Cer [NP] of the skin group with a low barrier function has a significantly reduced composition ratio relative to the total amount of ceramide compared to the normal skin group. Furthermore, Cer [AS] of the skin group with a low barrier function has a significantly increased composition ratio with respect to the total amount of ceramide compared to the normal skin group. Furthermore, Cer [EOS] in the skin group with a low barrier function has a significantly reduced composition ratio with respect to the total amount of ceramide compared to the normal skin group. Furthermore, Cer [EOH] in the skin group with a low barrier function has a significantly reduced composition ratio with respect to the total amount of ceramide compared to the normal skin group.

  In the skin group with low barrier function, as shown in FIG. 15, the average carbon number of the molecular species included in Cer [NDS] and Cer [NH] in the ceramide class is significantly different from that in the normal skin group. Is different. Here, as described above, the average carbon number means a weighted average value that is calculated by using the abundance ratio of each molecular species as a weight for a plurality of types of molecular species having different carbon numbers. Specifically, the average carbon number of Cer [NDS] and Cer [NH] in the skin group with low barrier function are all significantly decreased as compared with the normal skin group.

  Furthermore, in the skin group with a low barrier function, as shown in FIGS. 16 and 17, the amount and composition ratio of the molecular species having 34 carbon atoms in Cer [NS] are significantly increased in the ceramide class. ing.

  In the dry skin group, as shown in FIG. 18, the composition ratio of molecular species having 34 carbon atoms in Cer [NS] is particularly increased in the ceramide class.

It is a block diagram which shows roughly the structure of the analysis system for acquiring lipid composition information. It is a characteristic view which shows an example of the multistage mass chromatogram output from the analysis system shown in FIG. In the experimental system shown in Example 1, it is a characteristic view which shows the result of having compared the composition ratio of the ceramide component in an atopic dermatitis patient group and a healthy subject group. In the experimental system shown in Example 1, it is a characteristic view which shows the result of having compared the composition ratio of the ceramide component in a psoriasis patient group and a healthy subject group. In the experimental system shown in Example 1, the characteristic figure which shows the result of having compared the weighted average value which weighted the existence ratio about the carbon number of the molecular species which comprise the ceramide class in an atopic dermatitis patient group and a healthy subject group It is. In the experimental system shown in Example 1, it is a characteristic figure which shows the result of having compared the weighted average value which weighted the existence ratio about carbon number of the molecular species which comprise the ceramide class in a psoriasis patient group and a healthy subject group. In the experimental system shown in Example 1, it is a characteristic figure which shows the result of having compared the quantity per 1 microgram protein of the molecular species of 34 carbon atoms among Cer [NS] in an atopic dermatitis patient group and a healthy subject group. In the experimental system shown in Example 1, it is a characteristic figure which shows the result of having compared the quantity per 1 microgram protein of the molecular species of 34 carbon atoms among Cer [NS] in a psoriasis patient group and a healthy subject group. In the experimental system shown in Example 1, the characteristic figure which shows the result of having compared the composition ratio with respect to the total amount of Cer [NS] of the molecular species of 34 carbon atoms among Cer [NS] in the atopic dermatitis patient group and the healthy subject group It is. In the experimental system shown in Example 1, it is a characteristic figure which shows the result of having compared the composition ratio with respect to Cer [NS] total amount of the molecular species of 34 carbon atoms among Cer [NS] in a psoriasis patient group and a healthy subject group. In the experimental system shown in Example 1, it is a characteristic figure which shows the result of having compared the composition ratio of the molecular species with one or more unsaturated bonds about the ceramide class in an atopic dermatitis patient group and a healthy subject group. In the experimental system shown in Example 1, it is a characteristic view which shows the result of having compared the composition ratio of the molecular species with one or more unsaturated bonds about the ceramide class in a psoriasis patient group and a healthy subject group. In the experimental system shown in Example 1, it is a characteristic view which shows the result of having compared the composition ratio of the ceramide component in a dry skin group and a normal skin group. In the experimental system shown in Example 1, it is a characteristic view which shows the result of having compared the composition ratio of the ceramide component in a skin group with a low barrier function, and a normal skin group. In the experimental system shown in Example 1, the characteristic figure which shows the result of having compared the weighted average value which weighted the existence ratio about carbon number of the molecular species which comprise the ceramide class in the skin group with a low barrier function, and a normal skin group It is. In the experimental system shown in Example 1, it is a characteristic figure which shows the result of having compared the quantity per 1 microgram protein of the molecular species of carbon number 34 among Cer [NS] in a skin group with a low barrier function, and a normal skin group. In the experimental system shown in Example 1, the characteristic figure which shows the result of having compared the composition ratio with respect to Cer [NS] total amount of the molecular species of 34 carbon atoms among Cer [NS] in a skin group with a low barrier function, and a normal skin group. It is. In the experimental system shown in Example 1, it is a characteristic view which shows the result of having compared the composition ratio for every total carbon number of Cer [NS] in a dry skin group and a normal skin group.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... System 10 which performs analysis of lipid in skin stratum corneum ... Liquid chromatograph 11a, 11b ... High pressure gradient pump 12 ... Auto injector 13 ... Guard column 14 ... Separation column 20 ... Ionization promotion liquid feeding device 21 ... Pump 22 ... Tee connector 30 ... Mass spectrometer 31 ... Ionizer 32 ... Mass separation detector 40 ... Calculators a, b ... Eluent c ... Ionization accelerator d ... Lipid sample solution

Claims (16)

Among the lipid composition information obtained from subjects, N-acyl-sphingosine / ceramide component, N-acyl-6-OH-sphingosine / ceramide component, N- (α-OH-acyl) sphingosine / ceramide component, N- Composition of at least one component selected from the group consisting of (ω-OH-acyl) acyl-6-OH-sphingosine-ceramide component and N- (ω-OH-acyl) acyl-phytosphingosine-ceramide component with respect to the total amount of ceramide The ratio is compared with a reference value calculated based on lipid composition information obtained from healthy individuals and lipid composition information obtained from individuals with atopic dermatitis,
When the composition ratio of the N-acyl-sphingosine / ceramide component is equal to or higher than the reference value, when the composition ratio of the N-acyl-6-OH-sphingosine / ceramide component is less than the reference value, the N- (α- When the composition ratio of the OH-acyl) sphingosine / ceramide component is equal to or higher than the reference value, the composition ratio of the N- (ω-OH-acyl) acyl-6-OH-sphingosine / ceramide component is less than the reference value. And when the composition ratio of the N- (ω-OH-acyl) acyl-phytosphingosine / ceramide component is less than the reference value, and when at least one abnormal value is detected, the subject is atopic. A skin quality determination method for determining that there is a tendency for dermatitis.   The composition ratio of the N-acyl-sphingosine / ceramide component is 10% by weight as a reference value and an abnormal value when the reference value is equal to or greater than the reference value. The composition ratio of the N-acyl-6-OH-sphingosine / ceramide component 19% by weight as a reference value and an abnormal value when less than the reference value, and the composition ratio of the N- (α-OH-acyl) -sphingosine / ceramide component is 6% by weight as the reference value. When the value is equal to or higher than the value, an abnormal value is assumed, and the composition ratio of the N- (ω-OH-acyl) acyl-6-OH-sphingosine / ceramide component is less than the reference value with 1.5% by weight as a reference value. The composition ratio of the N- (ω-OH-acyl) acyl-phytosphingosine / ceramide component is 0.5% by weight as a reference value. Skin quality determination method according to claim 1, characterized in that an abnormal value in the case of less than the reference value. Among the lipid composition information obtained from subjects, N-acyl-dihydrosphingosine-ceramide component, N-acyl-sphingosine-ceramide component, N-acyl-6-OH-sphingosine-ceramide component and N- (α-OH -Acyl) -6-OH-sphingosine / ceramide For at least one component selected from the group consisting of components, the weighted average of the average carbon number of the molecular species contained in the component, with the abundance ratio of each molecular species as a weight Calculated as a value, and compared with a reference value calculated based on lipid composition information obtained from healthy individuals and lipid composition information obtained from individuals with atopic dermatitis ,
When the weighted average value of the carbon number of the N-acyl-dihydrosphingosine / ceramide component is less than the reference value, or when the weighted average value of the carbon number of the N-acyl-sphingosine / ceramide component is less than the reference value The weight average value of the carbon number of the N-acyl-6-OH-sphingosine-ceramide component is less than the reference value, and the N- (α-OH-acyl) -6-OH-sphingosine-ceramide component Skin quality determination , wherein when the weighted average value of the carbon number is less than the reference value, an abnormal value is detected, and when at least one abnormal value is detected, the subject determines that the subject has a tendency to atopic dermatitis Method. Regarding the weighted average value of the carbon number of the N-acyl-dihydrosphingosine / ceramide component, the reference value is 46.5 and an abnormal value is obtained when the reference value is less than the reference value. For the weighted average value of carbon number, the reference value is 46, and when it is less than the reference value, it is an abnormal value. For the weighted average value of the carbon number of the N-acyl-6-OH-sphingosine / ceramide component, the reference value 44 is an abnormal value when it is less than the reference value, and the weighted average value of the carbon number of the N- (α-OH-acyl) -6-OH-sphingosine / ceramide component is 44 reference values. The skin quality determination method according to claim 3, wherein an abnormal value is set when the value is less than the reference value. Among the lipid composition information obtained from the subject, the N-acyl-6-OH-sphingosine / ceramide component and / or the N- (ω-OH-acyl) acyl-sphingosine / ceramide component has one or more unsaturated bonds. Compare the ratio of many molecular species with the reference value calculated based on the lipid composition information obtained from healthy individuals and the lipid composition information obtained from individuals with atopic dermatitis ,
When the ratio of the N-acyl-6-OH-sphingosine / ceramide component is not less than the reference value, and the ratio of the N- (ω-OH-acyl) acyl-sphingosine / ceramide component is not less than the reference value. A skin quality determination method that determines that the subject has a tendency to atopic dermatitis when an abnormal value is detected and at least one abnormal value is detected . About the said ratio of the said N-acyl-6-OH- sphingosine and a ceramide component, when it is 1 weight% or more and the said reference value or more, it is set as an abnormal value, 6. The skin quality determination method according to claim 5, wherein the ratio of the ceramide component is determined to be an abnormal value when 20% by weight or more is a reference value and the reference value or more. Among the lipid composition information obtained from the subject, the total N-acyl-sphingosine / ceramide component in the total N-acyl-sphingosine / ceramide component and / or the total amount of all N-acyl-sphingosine / ceramide components per 34 g of protein of the molecular species having 34 carbon atoms. Comparing the composition ratio with a reference value calculated based on lipid composition information obtained from healthy individuals and lipid composition information obtained from individuals with atopic dermatitis ,
When the amount per 1 μg of protein of the molecular species having 34 carbon atoms is equal to or higher than the reference value, and when the composition ratio of the molecular species having 34 carbon atoms is equal to or higher than the reference value, an abnormal value is set, and at least one abnormal value The skin quality determination method of determining that the said subject has the tendency for atopic dermatitis when a value is detected . The amount per 1 μg protein of the molecular species having 34 carbon atoms is 0.04 (ng) as a reference value, and an abnormal value is obtained when the amount is equal to or higher than the reference value, and the composition ratio of the molecular species having 34 carbon atoms is the reference value. The skin quality determination method according to claim 7, wherein an abnormal value is set when the value is 2.5% by weight or more than the reference value. Selected from the group consisting of N-acyl-sphingosine-ceramide component, N-acyl-phytosphingosine-ceramide component and N- (α-OH-acyl) -sphingosine-ceramide component in the lipid composition information obtained from the subject Comparing the composition ratio with respect to the total amount of ceramide for at least one component to be compared with a reference value calculated based on lipid composition information obtained from a healthy individual and lipid composition information obtained from an individual with psoriasis,
The composition ratio of the N-acyl-sphingosine / ceramide component is an abnormal value when the reference value is 15% by weight or more, and the composition ratio of the N-acyl-phytosphingosine / ceramide component is a criterion. When the value is less than the reference value when the value is 12% by weight, an abnormal value is obtained. For the composition ratio of the N- (α-OH-acyl) -sphingosine / ceramide component, the reference value is 10% by weight. In case
The skin quality determination method of determining that the said subject has the tendency of psoriasis when at least 1 or more abnormal value is detected. Among the lipid composition information obtained from subjects, N-acyl-dihydrosphingosine-ceramide component, N-acyl-sphingosine-ceramide component, N-acyl-6-OH-sphingosine-ceramide component and N- (α-OH -Acyl) -6-OH-sphingosine / ceramide For at least one component selected from the group consisting of components, the weighted average of the average carbon number of the molecular species contained in the component, with the abundance ratio of each molecular species as a weight Calculated as a value, and compared with the lipid composition information obtained from healthy individuals and the lipid composition information obtained from individuals with psoriasis ,
When the weighted average value of the carbon number of the N-acyl-dihydrosphingosine / ceramide component is less than the reference value, the weighted average value of the carbon number of the N-acyl sphingosine / ceramide component is less than the reference value, When the weighted average value of the carbon number of the N-acyl-6-OH-sphingosine / ceramide component is less than the reference value, and of the N- (α-OH-acyl) -6-OH-sphingosine / ceramide component A skin quality determination method in which when the weighted average value of the carbon number is less than the reference value, an abnormal value is determined, and when at least one abnormal value is detected, the subject is determined to have a psoriasis tendency . Regarding the weighted average value of the carbon number of the N-acyl-dihydrosphingosine / ceramide component, the reference value is 45, and an abnormal value is obtained when the reference value is less than the reference value. As for the weighted average value, 44 reference values are set as abnormal values when the reference value is less than the reference value. For the weighted average value of the carbon number of the N-acyl-6-OH-sphingosine / ceramide component, 44 reference values are set. As an abnormal value when it is less than the reference value, the weighted average value of the carbon number of the N- (α-OH-acyl) -6-OH-sphingosine / ceramide component is 43.5 reference values. The skin quality determination method according to claim 10, wherein an abnormal value is set when the value is less than the reference value. Among the lipid composition information obtained from the subject, N-acyl-dihydrosphingosine and ceramide components, N-acyl-sphingosine and ceramide components, N-acyl-6-OH-sphingosine and ceramide components, N- (α-OH -Acyl) -sphingosine-ceramide component, N- (α-OH-acyl) -6-OH-sphingosine-ceramide component, N- (α-OH-acyl) -phytosphingosine-ceramide component and N- (ω-OH -Acyl) For at least one component selected from the group consisting of acyl-sphingosine and ceramide components, the ratio of molecular species having one or more unsaturated bonds was obtained from the lipid composition information obtained from healthy individuals and from psoriasis individuals. Compare with the reference value calculated based on the lipid composition information ,
When the ratio of the N-acyl-dihydrosphingosine / ceramide component is not less than the reference value, the ratio of the N-acyl-sphingosine / ceramide component is not less than the reference value, and the N-acyl-6-OH- When the ratio of the sphingosine / ceramide component is equal to or higher than the reference value, the N- (α-OH-acyl) when the ratio of the N- (α-OH-acyl) -sphingosine / ceramide component is higher than the reference value. ) -6-OH-sphingosine / ceramide component when the ratio is equal to or higher than the reference value, the N- (α-OH-acyl) -phytosphingosine / ceramide component is equal to or higher than the reference value, and When the ratio of the N- (ω-OH-acyl) acyl-sphingosine / ceramide component is equal to or higher than the reference value, an abnormal value is set. If it detects more abnormal values, the subject is judged that there is a tendency of psoriasis, skin type determination method. The above-mentioned ratio of the N-acyl-dihydrosphingosine / ceramide component is an abnormal value when the reference value is 1% by weight or more, and the above-mentioned ratio of the N-acyl-sphingosine / ceramide component is the reference value. 1% by weight is an abnormal value if it is above the reference value, and the above ratio of the N-acyl-6-OH-sphingosine / ceramide component is an abnormal value if the reference value is 1% by weight or more. The ratio of the N- (α-OH-acyl) -sphingosine / ceramide component is defined as an abnormal value when the reference value is 1% by weight or more, and the N- (α-OH-acyl)- The above ratio of the 6-OH-sphingosine / ceramide component is defined as an abnormal value when the reference value is 1% by weight or more, and the above N- (α The above ratio of the OH-acyl) -phytosphingosine / ceramide component is defined as an abnormal value when the reference value is 1% by weight or more, and the N- (ω-OH-acyl) acyl-sphingosine / ceramide component skin quality determination method of claim 12, characterized in that an abnormal value in the case of more than the reference value the reference value as a 20 wt% for the ratio. Among the lipid composition information obtained from the subject, the composition ratio of the total N-acyl-sphingosine / ceramide component to the amount of the molecular species having 34 carbon atoms and / or the total amount of the total N-acyl-sphingosine / ceramide component, Compared with the reference value calculated based on the lipid composition information obtained from healthy individuals and the lipid composition information obtained from individuals with psoriasis ,
An abnormal value was detected when the amount of the molecular species having 34 carbon atoms was equal to or greater than the reference value, and when the composition ratio of the molecular species having 34 carbon atoms was equal to or greater than the reference value, and at least one abnormal value was detected. In the case, the skin quality determination method of determining that the subject is prone to psoriasis . Regarding the amount of the molecular species having 34 carbon atoms, 0.07 (ng) per 1 μg of protein is used as a reference value, and an abnormal value is obtained when the amount exceeds the reference value. The composition ratio of the molecular species having 34 carbon atoms is the reference value. The skin quality determination method according to claim 14, wherein an abnormal value is set when the value is 3.0% by weight or more than the reference value.   The lipid composition information is data generated by separating and detecting a lipid sample collected from the subject by liquid chromatography-mass spectrometry and analyzing the detection data by a multistage mass chromatogram. The skin quality determination method according to any one of claims 1 to 15.

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