Patent Publication Number: US-2019170683-A1

Title: Thin layer chromatography plate and sample analysis method using same

Description:
TECHNICAL FIELD 
     The present disclosure relates to a thin layer chromatography plate and a sample analysis using the same. 
     BACKGROUND ART 
     Chromatography and electrophoresis, for example, have been known as a method for separating a specific component from a mixture containing multiple components. Thin layer chromatography which is a kind of chromatography techniques makes it possible to easily and quickly separate multiple components from each other. 
     As shown in  FIG. 15 , PTL 1 discloses thin layer chromatography plate  2000  provided with first separating agent layer  2031  and second separating agent layer  2032 . Second separating agent layer  2032  is adjacent to first separating agent layer  2031 . First separating agent layer  2031  and second separating agent layer  2032  are respectively formed from separating agents having different optical responses. 
     When thin layer chromatography plate  2000  is used, multiple components can be separated from each other as described below. Sample  2060  is placed on first separating agent layer  2031  and developed in direction X. Then, second separating agent layer  2032  is dried. Next, the orientation of thin layer chromatography plate  2000  is changed, and sample  2060  is developed in direction Y orthogonal to direction X. The multiple components are separated from each other in second separating agent layer  2032 . 
     CITATION LIST 
     Patent Literature 
     PTL 1: WO 2011/149041 A 
     SUMMARY OF THE INVENTION 
     According to the method disclosed in PTL 1, it is necessary that, after the sample is developed in first separating agent layer  2031 , second separating agent layer  2032  is dried. 
     The present disclosure aims to provide a technique for separating multiple components from each other more easily and more quickly. 
     Specifically, the present disclosure provides a thin layer chromatography plate described below. The thin layer chromatography plate includes a substrate and a separation layer disposed on the substrate, the sepatatgion layer separating multiple components included in a sample from each other. The separation layer has a first layer that has a band shape and extends in a first development direction and a second layer that extends in a second development direction orthogonal to the first development direction. The second layer is in contact with the first layer, the first layer includes a hydrophilic porous body, and the second layer includes a hydrophobic porous body. 
     According to the thin layer chromatography plate in the present disclosure, multiple components can be separated from each other more easily and more quickly. 
    
    
     
       BRIEF DESCRIPTION OF DRAWINGS 
         FIG. 1A  is a plan view of a thin layer chromatography plate according to a first exemplary embodiment of the present disclosure. 
         FIG. 1B  is a sectional view of the thin layer chromatography plate shown in  FIG. 1A  along second development direction Y. 
         FIG. 2A  is a view showing a state where a sample is placed on the thin layer chromatography plate according to the first exemplary embodiment of the present disclosure. 
         FIG. 2B  is a view showing a state where the thin layer chromatography plate shown in  FIG. 2A  is brought into contact with a first developing solvent. 
         FIG. 2C  is a view showing a state where the thin layer chromatography plate shown in  FIG. 2B  is brought into contact with a second developing solvent by changing the orientation of the thin layer chromatography plate. 
         FIG. 3A  is a plan view of a thin layer chromatography plate according to a second exemplary embodiment of the present disclosure. 
         FIG. 3B  is a sectional view of the thin layer chromatography plate shown in  FIG. 3A  along second development direction Y. 
         FIG. 3C  is a sectional view of a thin layer chromatography plate along second development direction Y according to a modification of the second exemplary embodiment of the present disclosure. 
         FIG. 3D  is a sectional view of a thin layer chromatography plate along second development direction Y according to another modification of the second exemplary embodiment of the present disclosure. 
         FIG. 4A  is a plan view of a thin layer chromatography plate according to a third exemplary embodiment of the present disclosure. 
         FIG. 4B  is a sectional view of the thin layer chromatography plate shown in  FIG. 4A  along second development direction Y. 
         FIG. 5A  is a plan view of a thin layer chromatography plate according to a fourth exemplary embodiment of the present disclosure. 
         FIG. 5B  is a sectional view of the thin layer chromatography plate shown in  FIG. 5A  along line VB-VB. 
         FIG. 6A  is a view showing a state where a sample is placed on the thin layer chromatography plate according to the fourth exemplary embodiment of the present disclosure. 
         FIG. 6B  is a view showing a state where a voltage is applied to a pair of electrodes in the thin layer chromatography plate shown in  FIG. 6A . 
         FIG. 6C  is a view showing a state where the thin layer chromatography plate shown in  FIG. 6B  is brought into contact with a second developing solvent. 
         FIG. 7A  is a plan view of a thin layer chromatography plate according to a fifth exemplary embodiment of the present disclosure. 
         FIG. 7B  is a sectional view of the thin layer chromatography plate shown in  FIG. 7A  along second development direction Y. 
         FIG. 8A  is a plan view of a thin layer chromatography plate according to a sixth exemplary embodiment of the present disclosure. 
         FIG. 8B  is a sectional view of the thin layer chromatography plate shown in  FIG. 8A  along second development direction Y. 
         FIG. 9A  is a view showing a state where a sample is placed on the thin layer chromatography plate according to the sixth exemplary embodiment of the present disclosure. 
         FIG. 9B  is a view showing a state where the thin layer chromatography plate shown in  FIG. 9A  is brought into contact with a first developing solvent. 
         FIG. 9C  is a view showing a state where the thin layer chromatography plate shown in  FIG. 9B  is brought into contact with a second developing solvent by changing the orientation of the thin layer chromatography plate. 
         FIG. 10A  is a plan view of a thin layer chromatography plate according to a seventh exemplary embodiment of the present disclosure. 
         FIG. 10B  is a sectional view of the thin layer chromatography plate shown in  FIG. 10A  along second development direction Y. 
         FIG. 10C  is a sectional view of a thin layer chromatography plate along second development direction Y according to a modification of the seventh exemplary embodiment of the present disclosure. 
         FIG. 10D  is a sectional view of a thin layer chromatography plate along second development direction Y according to another modification of the seventh exemplary embodiment of the present disclosure. 
         FIG. 11A  is a plan view of a thin layer chromatography plate according to an eighth exemplary embodiment of the present disclosure. 
         FIG. 11B  is a sectional view of the thin layer chromatography plate shown in  FIG. 11A  along second development direction Y. 
         FIG. 12A  is a plan view of a thin layer chromatography plate according to a ninth exemplary embodiment of the present disclosure. 
         FIG. 12B  is a sectional view of the thin layer chromatography plate shown in  FIG. 12A  along line XIIB-XIIB. 
         FIG. 13A  is a view showing a state where a sample is placed on the thin layer chromatography plate according to the ninth exemplary embodiment of the present disclosure. 
         FIG. 13B  is a view showing a state where a voltage is applied to a pair of electrodes in the thin layer chromatography plate shown in  FIG. 13A . 
         FIG. 13C  is a view showing a state where the thin layer chromatography plate shown in  FIG. 13B  is brought into contact with a second developing solvent. 
         FIG. 14A  is a plan view of a thin layer chromatography plate according to a tenth exemplary embodiment of the present disclosure. 
         FIG. 14B  is a sectional view of the thin layer chromatography plate shown in  FIG. 14A  along second development direction Y. 
         FIG. 14C  is a sectional view of a thin layer chromatography plate along second development direction Y according to a modification of the tenth exemplary embodiment of the present disclosure. 
         FIG. 14D  is a sectional view of a thin layer chromatography plate along second development direction Y according to another modification of the tenth exemplary embodiment of the present disclosure. 
         FIG. 15  is a plan view of a conventional thin layer chromatography plate. 
     
    
    
     DESCRIPTION OF EMBODIMENTS 
     Underlying Knowledge of the Present Disclosure 
     A human skin state can be checked by analyzing proteins included in skin. The protein analysis is conducted in the manner described below, for example. A sample such as a surface skin is extracted from the skin of a subject. The sample includes a plural kinds of proteins. The multiple proteins included in the sample are separated from each other using thin layer chromatography. Each of the separated proteins is identified. 
     For example, if the sample includes a protein related to by rough skin, it is found that the subject has rough skin If the skin state of the subject can be recognized, cosmetics suitable for the subject can be recommended. It is convenient to check the skin state of the subject and recommend cosmetics based on the check result in cosmetics retail stores. When doing so, protein analysis needs to be quickly conducted during a waiting time of the subject. 
     A thin layer chromatography plate according to a first aspect of the present disclosure has the configurations described below. 
     Specifically, the thin layer chromatography plate includes: a substrate; and a separation layer disposed on the substrate for separating multiple components included in a sample from each other. The separation layer has a first layer that has a band shape and extends in a first development direction and a second layer that extends in a second development direction orthogonal to the first development direction. The second layer is in contact with the first layer, the first layer includes a hydrophilic porous body, and the second layer includes a hydrophobic porous body. 
     According to the first aspect, the second layer in the separation layer includes a hydrophobic porous body, whereby water is difficult to penetrate into the second layer. That is, if a developing solvent is selected as appropriate, pores in the porous body constituting the second layer hardly contains the developing solvent, after the multiple components included in the sample are developed in the first development direction. Therefore, it is unnecessary to dry the second layer after the multiple components are developed in the first development direction. Thus, the multiple components can be separated from each other more easily and more quickly. 
     According to a second aspect of the present disclosure, the first layer and the second layer of the thin layer chromatography plate according to the first aspect are both disposed on the substrate, and a lateral surface of the first layer is in contact with a lateral surface of the second layer, for example. According to the second aspect, the developing solvent can be easily moved from the first layer to the second layer. 
     According to a third aspect of the present disclosure, the separation layer of the thin layer chromatography plate according to the first aspect further includes a third layer that is in contact with the second layer, and the first layer, the second layer, and the third layer are arrayed in sequence in the second development direction, for example. In this configuration, the third layer includes a porous body. Further, at least one requirement selected from among a requirement of a composition of the third layer being different from a composition of the second layer and a requirement of a structure of the third layer being different from a structure of the second layer is satisfied. 
     According to the third aspect, the second layer in the separation layer includes a hydrophobic porous body, whereby water is difficult to penetrate into the second layer. That is, if a developing solvent is selected as appropriate, pores in the porous body constituting the second layer hardly contains the developing solvent, after the multiple components included in the sample are developed in the first development direction. Therefore, it is unnecessary to dry the second layer after the multiple components are developed in the first development direction. Thus, the multiple components can be separated from each other more easily and more quickly. The third layer in the separation layer induces an interaction different from the interaction induced by the second layer, with respect to the multiple components included in the sample. Therefore, the multiple components which are not separated from each other in the second layer are separated from each other in the third layer. 
     According to a fourth aspect of the present disclosure, the first layer, the second layer, and the third layer of the thin layer chromatography plate according to the third aspect are disposed on the substrate, and a lateral surface of the first layer and a lateral surface of the second layer are in contact with each other, for example. According to the fourth aspect, the developing solvent can be easily moved from the first layer to the second layer. 
     According to a fifth aspect of the present disclosure, the separation layer of the thin layer chromatography plate according to the second aspect or the fourth aspect further includes a functional layer that extends in the second development direction and includes the hydrophobic porous body or another hydrophobic porous body, for example. The functional layer is in contact with the first layer, and the functional layer, the first layer, and the second layer are arrayed in sequence in the second development direction. According to the fifth aspect, a gradient occurring in a movement distance of the developing solvent in the second development direction is reduced in the functional layer. Thus, the multiple components can move straight in the second development direction in the second layer. 
     According to a sixth aspect of the present disclosure, the second layer of the thin layer chromatography plate according to the first aspect or the third aspect is disposed on the substrate, the first layer is disposed on the second layer, and a lower surface of the first layer and an upper surface of the second layer are in contact with each other, for example. According to the sixth aspect, a gradient occurring in a movement distance of the developing solvent in the second development direction is reduced in a part of the second layer. Thus, the multiple components can move straight in the second development direction in the second layer. 
     According to the seventh aspect of the present disclosure, the first layer of the thin layer chromatography plate according to the sixth aspect is located between one end and the other end of the second layer in the second development direction, for example. According to the seventh aspect, the multiple components can move straight in the second development direction in the second layer. 
     According to an eighth aspect of the present disclosure, the hydrophobic porous body of the thin layer chromatography plate according to any one of the first to seventh aspects is an aggregate of silica gel particles modified with a hydrophobic functional group, for example. According to the eighth aspect, it is unnecessary to dry the second layer after the multiple components included in the sample are developed in the first development direction. Thus, the multiple components can be separated from each other more easily and more quickly. 
     According to a ninth aspect of the present disclosure, the thin layer chromatography plate according to any one of the first to eighth aspects further includes a pair of electrodes disposed at both ends of the first layer in the first development direction, for example. According to the ninth aspect, electrophoresis of the multiple components included in the sample can be achieved. 
     A sample analysis method according to a tenth aspect of the present disclosure includes the following steps. Specifically, the sample analysis method includes: placing a sample onto a first layer of the thin layer chromatography plate according to any one of the first to eighth aspects; and bringing an end of the first layer in the first development direction into contact with a first developing solvent. The sample analysis method further includes changing the orientation of the thin layer chromatography plate to bring the thin layer chromatography plate into contact with a second developing solvent containing an organic solvent with the first layer being impregnated with the first developing solvent. 
     According to the tenth aspect, due to the second layer in the separation layer including the hydrophobic porous body, pores in the porous body constituting the second layer hardly contains the first developing solvent, after the multiple components included in the sample are developed in the first development direction. Therefore, it is unnecessary to dry the second layer after the multiple components included in the sample are developed in the first development direction. Thus, the multiple components can be separated from each other more easily and more quickly. 
     According to an eleventh aspect of the present disclosure, the first developing solvent used in the sample analysis method according to the tenth aspect is an aqueous solution, for example. According to the eleventh aspect, it is unnecessary to dry the second layer after the multiple components included in the sample are developed in the first development direction. Thus, the multiple components can be separated from each other more easily and more quickly. 
     According to a twelfth aspect of the present disclosure, the second developing solvent used in the sample analysis method according to the tenth or eleventh aspect is a mixed solvent containing the organic solvent and water, for example. According to the tenth aspect, the multiple components included in the sample can be easily dissolved in the second developing solvent. 
     A sample analysis method according to a thirteenth aspect of the present disclosure includes the following steps. Specifically, the sample analysis method includes: placing a sample onto the first layer of the thin layer chromatography plate according to the ninth aspect; applying a voltage to the pair of electrodes; and bringing the thin layer chromatography plate into contact with a developing solvent containing an organic solvent. 
     According to the eleventh aspect, due to the second layer in the separation layer including the hydrophobic porous body, pores in the porous body constituting the second layer hardly contains the developing solvent, after the multiple components included in the sample are developed in the first development direction. Therefore, it is unnecessary to dry the second layer after the multiple components included in the sample are developed in the first development direction. Thus, the multiple components can be separated from each other more easily and more quickly. 
     Exemplary embodiments of the present disclosure will be described below with reference to the drawings. The present disclosure is not limited to the following exemplary embodiments. 
     First Exemplary Embodiment 
     As shown in  FIGS. 1A and 1B , thin layer chromatography plate  100  (hereinafter referred to as “TLC plate  100 ”) according to the present first exemplary embodiment has substrate  10  and separation layer  20 . Substrate  10  has a plate shape, for example. Substrate  10  has, for example, a rectangular shape in a plan view. Separation layer  20  is disposed on substrate  10 . Separation layer  20  covers the surface of substrate  10 . Substrate  10  has two pairs of end faces facing each other. In the present exemplary embodiment, first development direction X extends from one end face of one of two pairs of end faces of substrate  10  to the other end face. Multiple components included in the sample are developed in first development direction X in a first stage. Second development direction Y extends from one end face of the other of two pairs of end faces of substrate  10  to the other end face. Multiple components included in the sample are developed in second development direction Y in a second stage. Second development direction Y is orthogonal to first development direction X. 
     Separation layer  20  separates the multiple components included in the sample from each other. Separation layer  20  includes first layer  31  and second layer  32 . First layer  31  is a band-shaped layer. First layer  31  has a rectangular band shape in a plan view. First layer  31  extends in first development direction X. First layer  31  extends from one (end face  91 ) of a pair of end faces (end face  91  and end face  92 ) of substrate  10  to the other (end face  92 ) in first development direction X. Note that first layer  31  may not extend to the other end face (end face  92 ) of the pair of end faces of substrate  10  in first development direction X. 
     Second layer  32  has a rectangular shape in a plan view. Second layer  32  extends in second development direction Y. Second layer  32  is in contact with first layer  31 . When separation layer  20  is viewed in a plan view, one side (long side) of first layer  31  is in contact with one side of second layer  32 . The length of one side (long side) of first layer  31  is equal to the length of one side of second layer  32 . First layer  31  and second layer  32  constitute boundary face  40 . Boundary face  40  extends in first development direction X. Second layer  32  extends from boundary face  40  to end face  93  of substrate  10  in second development direction Y. Note that second layer  32  may not extend to end face  93  of substrate  10  in second development direction Y. 
     In the present exemplary embodiment, first layer  31  and second layer  32  are both disposed on substrate  10 . In other words, first layer  31  and second layer  32  are both in contact with substrate  10 . A lateral surface of first layer  31  and a lateral surface of second layer  32  are in contact with each other. When the multiple components are developed in second development direction Y, a developing solvent can easily move from first layer  31  to second layer  32  through boundary face  40 . 
     The material of substrate  10  is not particularly limited, as long as it can maintain the shape of TLC plate  100  without being eluted in a developing solvent. The material of substrate  10  is glass, resin, metal, or paper, for example. Substrate  10  is typically a glass plate or an aluminum film. 
     First layer  31  includes a hydrophilic porous body. In the present specification, “being hydrophilic” means that, when water is brought into contact with the porous body, water can move in the porous body at a rate of 5 mm/min or higher due to capillary force, for example. When water is selected as the developing solvent, first layer  31  can carry water from one end to the other end of first layer  31  in first development direction X due to capillary force. The hydrophilic porous body is not particularly limited. The material of the hydrophilic porous body includes at least one selected from the group consisting of a fiber material, and an inorganic material and a polymer material which are hydrophilic, for example. 
     The fiber material includes at least one selected from the group consisting of a plant fiber, an animal fiber, a recycled fiber, a synthetic fiber, and a glass fiber, for example. The plant fiber includes cellulose, for example. The synthetic fiber includes cellulose acetate, for example. 
     The hydrophilic polymer material includes at least one selected from the group consisting of agarose, dextran, and mannan, for example. The inorganic material includes at least one selected from the group consisting of alumina, silicon dioxide, and zirconia, for example. 
     The hydrophilic porous body is filter paper, for example. The hydrophilic porous body is an aggregate of at least one kind of inorganic particles selected from the group consisting of alumina particles, silica gel particles, silicon pillar, zeolite particles, diatomaceous earth, and zirconia particles, for example. 
     An average pore diameter of first layer  31  may be within a range from 0.01 μm to 100 μm. When the hydrophilic porous body is an aggregate of inorganic particles, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. The “average pore diameter” can be measured with the following method. Specifically, the surface or cross-section of first layer  31  is observed with an electron microscope (for example, an scanning electron microscope). Pore diameters of a plurality of observed pores (for example, random 50 pores) are measured. The average pore diameter is determined based on the average value calculated using the measured values. The diameter of a circle having an area equal to the area of the pore observed with the electron microscope can be regarded as the pore diameter. The “average particle diameter” can be measured with the following method. Specifically, the surface or cross-section of first layer  31  is observed with an electron microscope, and diameters of random number (for example, 50) of particles constituting first layer  31  are measured. The average particle diameter is determined based on the average value calculated using the obtained measured values. The diameter of a circle having an area equal to the area of the particle observed with the electron microscope can be regarded as the particle diameter. 
     First layer  31  may further include an additive. Examples of the additive include a fluorescence indicator, a binder, and a metal oxide. 
     Examples of the fluorescence indicator include magnesium tungstate and zinc silicate containing manganese. When first layer  31  includes the fluorescence indicator, positions of the multiple components can be detected by irradiating first layer  31  with ultraviolet ray. 
     The binder includes at least one selected from the group consisting of an inorganic binder, an organic fiber, a thickener, and an organic binder, for example. Examples of the inorganic binder include plaster and colloidal silica. Examples of the organic fiber include microfibrillar cellulose. Examples of the thickener include hydroxyethyl cellulose and carboxymethyl cellulose. Examples of the organic binder include polyvinyl alcohol and polyacrylic acid. When first layer  31  includes the binder, adhesiveness between substrate  10  and first layer  31  is improved. When the porous body of first layer  31  is an aggregate of inorganic particles, durability of the aggregate of inorganic particles is improved due to the binder. 
     The metal oxide includes at least one selected from the group consisting of titanium oxide, aluminum oxide, tin oxide, zinc oxide, tungsten oxide, manganese oxide, nickel oxide, copper oxide, and magnesium oxide, for example. The metal oxide may be charged when getting wet with the developing solvent. Therefore, when first layer  31  includes the metal oxide, the interaction between the multiple components included in the sample and first layer  31  varies. Thus, the multiple components may be easily separated from each other in first layer  31 . 
     The above-mentioned additives may be mixed into the porous body constituting first layer  31 . When the additive is a binder or a metal oxide, the additive may cover the surface of the porous body. The additive may coat the surfaces of inorganic particles constituting the porous body. 
     Second layer  32  includes a hydrophobic porous body. In the present specification, “being hydrophobic” means that, when water is brought into contact with the porous body, water can move in the porous body at a rate lower than 5 mm/min due to capillary force, or water does not move inside the porous body due to capillary force, for example. When water moves inside the hydrophobic porous body, the movement rate of water may be lower than 1 mm/min. When second layer  32  is brought into contact with water, water hardly penetrates into second layer  32 . When the developing solvent contains an organic solvent, second layer  32  can carry the developing solvent from one end to the other end of second layer  32  in second development direction Y due to capillary force. The hydrophobic porous body is not particularly limited. The material of the hydrophobic porous body is, for example, a hydrophobic polymer material. The hydrophobic polymer material includes at least one selected from the group consisting of fluororesin, polystyrene, polyethylene, and polypropylene, for example. 
     The hydrophobic porous body is, for example, a porous body of a hydrophobic polymer material or an aggregate of hydrophobic polymer material particles. The hydrophobic porous body is an aggregate of inorganic particles modified with a hydrophobic functional group, for example. The hydrophobic functional group includes a functional group having a hydrocarbon group at the end, for example. The hydrocarbon group includes at least one selected from the group consisting of an octadecyl group, an octyl group, a t-butyl group, a trimethylsilyl group, and a phenyl group, for example. The inorganic particles include at least one kind selected from the group consisting of alumina particles, silica gel particles, silicon pillar, zeolite particles, diatom earth, and zirconia particles, for example. The hydrophobic porous body is typically an aggregate of silica gel particles modified with a hydrophobic functional group. Whether the inorganic particles are modified with a hydrophobic functional group can be confirmed by conducting an elemental analysis on the cross-section of second layer  32 , for example. The elemental analysis can be conducted by X-ray photoelectron spectroscopy (XPS) or energy dispersive X-ray spectroscopy (EDX), for example. 
     An average pore diameter of second layer  32  may be within a range from 0.01 μm to 100 μm. When the hydrophobic porous body is an aggregate of inorganic particles modified with a hydrophobic functional group, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. Second layer  32  may further include any of the above-mentioned additives. 
     Length L 1  of first layer  31  in first development direction X is not particularly limited. Length L 1  is determined according to the porous body constituting first layer  31 , a size of a container for housing TLC plate  100 , and the like. Length L 1  is 10 mm to 100 mm, for example. The length of second layer  32  and the length of substrate  10  in first development direction X are typically equal to length L 1 . 
     Length L 2  of first layer  31  in second development direction Y is not particularly limited. Length L 2  is determined according to an amount of the sample to be placed on first layer  31 , for example. As length L 2  is smaller, the multiple components are more easily separated from each other when the multiple components are developed in second development direction Y. Length L 2  is 0.5 mm to 10 mm, for example. 
     Length L 3  of second layer  32  in second development direction Y is not particularly limited. Length L 3  is determined according to the porous body constituting second layer  32 , a size of a container for housing TLC plate  100 , and the like. Length L 3  is 20 mm to 200 mm, for example. The length of substrate  10  in second development direction Y is typically equal to the total of length L 2  and length L 3 . 
     Thickness L 4  of first layer  31  is not particularly limited. Thickness L 4  is determined according to the porous body constituting first layer  31 , for example. Thickness L 4  is 0.05 mm to 1 mm, for example. The thickness of second layer  32  is typically equal to thickness L 4  of first layer  31 . 
     Thickness L 5  of substrate  10  is not particularly limited as long as the shape of TLC plate  100  can be maintained. Thickness L 5  is 0.1 mm to 5 mm, for example. 
     Next, a manufacturing method of TLC plate  100  will be described. 
     First, a first dispersion liquid containing inorganic particles is prepared. The first dispersion liquid can be obtained by dispersing inorganic particles into a coating solvent. 
     The coating solvent includes at least one selected from the group consisting of water and an organic solvent, for example. The organic solvent includes at least one selected from the group consisting of alcohol, ketone, ether, nitrile, sulfoxide, sulfone, ester, carboxylic acid, amide, hydrocarbon, aromatic hydrocarbon, and halogen-containing compound, for example. 
     Examples of alcohol include methanol, ethanol, and isopropyl alcohol. Examples of ketone include acetone and ethyl methyl ketone. Examples of ether include tetrahydrofuran and dioxane. Examples of nitrile include acetonitrile. Examples of sulfoxide include dimethyl sulfoxide. Examples of sulfone include sulfolane. Examples of ester include ethyl acetate. Examples of carboxylic acid includes formic acid and acetic acid. Examples of amide include dimethylformamide. Examples of hydrocarbon include pentane and hexane. Examples of aromatic hydrocarbon include benzene, toluene, and xylene. Examples of halogen-containing compound include methylene chloride, chloroform, bromoform, chlorobenzene, and bromobenzene. 
     The first dispersion liquid is applied on a part of the surface of substrate  10  to form a coating film. The coating film is dried, whereby first layer  31  is formed on substrate  10 . When the hydrophilic porous body is filter paper, first layer  31  is formed on substrate  10  by bonding the hydrophilic porous body to a part of the surface of substrate  10  under pressure. 
     Next, a second dispersion liquid containing inorganic particles modified with a hydrophobic functional group is prepared. The second dispersion liquid can be obtained by dispersing inorganic particles modified with a hydrophobic functional group into a coating solvent. The materials mentioned above can be used for the coating solvent. 
     The second dispersion liquid is applied on a part of the surface of substrate  10  to form a coating film. The coating film is dried, whereby second layer  32  is formed on substrate  10 . When the hydrophobic porous body is a porous body of a hydrophobic polymer material, second layer  32  is formed on substrate  10  by bonding the hydrophobic porous body to a part of the surface of substrate  10  under pressure. 
     The second dispersion liquid may contain inorganic particles not modified with a hydrophobic functional group, in place of inorganic particles modified with a hydrophobic functional group. In such a case, second layer  32  is formed in the manner described below. The second dispersion liquid is applied on substrate  10  to form a coating film. The coating film is dried, whereby a untreated layer of second layer  32  is formed. A silane coupling agent having a hydrophobic functional group is applied on the untreated layer. The silane coupling agent is reacted with the inorganic particles included in the untreated layer. Thus, second layer  32  is formed on substrate  10 . The silane coupling agent may be applied on a coating film, not on the untreated layer. 
     The silane coupling agent is not particularly limited. The silane coupling agent may include at least one selected from the group consisting of dimethyloctadecylchlorosilane, dimethyloctylchlorosilane, t-butyldimethylchlorosilane, trimethylchlorosilane, and phenyldimethylchlorosilane. 
     First layer  31  and second layer  32  may be formed by the following method. The first dispersion liquid is applied on the entire surface of substrate  10  to form a coating film The coating film is dried, whereby a untreated layer of second layer  32  and first layer  31  are formed on substrate  10 . A silane coupling agent having a hydrophobic functional group is applied on the untreated layer of second layer  32 . The silane coupling agent is reacted with the inorganic particles. Thus, first layer  31  and second layer  32  are formed on substrate  10 . 
     The order of formation of first layer  31  and second layer  32  on substrate  10  is not particularly limited. First layer  31  may be formed on substrate  10  after second layer  32  is formed on substrate  10 . 
     Next, the sample analysis method using TLC plate  100  will be described. 
     First, sample  60  is placed on first layer  31  of separation layer  20  of TLC plate  100 , as shown in  FIG. 2A . When sample  60  is placed on first layer  31 , sample  60  penetrates into first layer  31 , so that circular spot  61  is formed. Sample  60  is an aqueous solution containing a plurality of proteins, for example. The content of the plurality of proteins in sample  60  is from 0.01 wt. % to 1 wt. %, for example. The volume of sample  60  placed on first layer  31  is 0.1 μL to 2 μL, for example. The position where sample  60  is to be placed on first layer  31  is not particularly limited, as long as sample  60  is not in direct contact with the first developing solvent and the second developing solvent. 
     Then, as shown in  FIG. 2B , TLC plate  100  is placed in container  75  with end  31   a  of first layer  31  in first development direction X being directed downward. Container  75  contains first developing solvent  70 . Container  75  is a glass jar, for example. Container  75  may be installed inside an analyzing device (not shown). 
     First developing solvent  70  is not particularly limited, as long as it does not move to the inside of second layer  32  when being brought into contact with the surface of second layer  32 . First developing solvent  70  is water or an aqueous solution, for example. A solute of the aqueous solution contains at least one selected from the group consisting of phosphate, citrate, acetate, and borate, for example. The aqueous solution may be a buffer solution such as a phosphate buffer solution, a tris buffer solution, a citrate buffer solution, an acetate buffer solution, or a borate buffer solution. In the present exemplary embodiment, first developing solvent  70  does not contain an organic solvent. However, first developing solvent  70  may contain an organic solvent. When first developing solvent  70  contains an organic solvent, first developing solvent  70  typically contains 80 volume % or more water. 
     When TLC plate  100  is placed in container  75 , end  31   a  of first layer  31  is in contact with first developing solvent  70 . The liquid level of first developing solvent  70  is set to prevent direct contact between first developing solvent  70  and sample  60 . Due to the capillary force, first developing solvent  70  moves in first development direction X from end  31   a  of first layer  31 . When first developing solvent  70  and sample  60  are brought into contact with each other, the multiple components included in sample  60  are dissolved into first developing solvent  70 . The multiple components dissolved in first developing solvent  70  move in first development direction X along with first developing solvent  70 . The multiple components move while repeatedly adsorbing and desorbing to and from the porous body constituting first layer  31 . The frequency of adsorption and desorption varies in each component, and thus, the multiple components are separated from each other in first layer  31 . Due to the development of sample  60  in first development direction X, spots  62 ,  63 ,  64 , and  65  are newly generated. Spots  62 ,  63 ,  64 , and  65  respectively indicate that any of the multiple components included in the sample is located therein. 
     Then, the orientation of TLC plate  100  is changed. The analyzing device may include a mechanism for changing the orientation of TLC plate  100 . As shown in  FIG. 2C , TLC plate  100  is placed in container  76  with end  31   b  of first layer  31  in second development direction Y being directed downward. Container  76  contains second developing solvent  71 . Container  76  is a glass jar, for example. Container  76  may be installed inside the analyzing device. 
     Second developing solvent  71  is not particularly limited, as long as it contains an organic solvent. Second developing solvent  71  contains an organic solvent, so that it can penetrate into second layer  32 . The materials mentioned above as examples of the coating solvent can be used as the organic solvent. The organic solvent contains at least one selected from the group consisting of methanol, ethanol, isopropyl alcohol, acetonitrile, and acetic acid, for example. When second developing solvent  71  contains carboxylic acid and the sample contains proteins, the frequency of absorption and desorption of proteins to and from the porous body constituting second layer  32  is improved. Second developing solvent  71  may contain the organic solvent in an amount of 20 wt. % or more. Second developing solvent  71  may contain water in addition to the organic solvent. That is, second developing solvent  71  may be a mixed solvent containing the organic solvent and water. When second developing solvent  71  contains water and the sample contains proteins, solubility of the proteins in second developing solvent  71  is improved. In other words, the multiple components included in the sample can be easily dissolved in second developing solvent  71 . Specific examples of second developing solvent  71  include a mixed solvent in which isopropyl alcohol, acetic acid, and water are mixed in a weight ratio of 40:5:55. 
     When TLC plate  100  is placed in container  76 , end  31   b  of first layer  31  is in contact with second developing solvent  71 . At that time, first layer  31  is impregnated with first developing solvent  70 . The liquid level of second developing solvent  71  is set to prevent direct contact between second developing solvent  71  and spots  62 ,  63 ,  64 , and  65 . Second developing solvent  71  penetrates into first layer  31 . Second developing solvent  71  moves from end  31   b  of first layer  31  in second development direction Y due to capillary force, along with first developing solvent  70  penetrating into first layer  31 . When second developing solvent  71  is brought into contact with the multiple components located in spots  62 ,  63 ,  64 , and  65 , the multiple components are dissolved into second developing solvent  71 . The multiple components dissolved in second developing solvent  71  move in second development direction Y along with second developing solvent  71 . The multiple components move while repeatedly adsorbing and desorbing to and from the porous body constituting second layer  32 . The multiple components which are not separated from each other in first layer  31  are separated from each other in second layer  32 . 
     Second layer  32  of TLC plate  100  includes a hydrophobic porous body. First developing solvent  70  hardly penetrates into second layer  32 . That is, pores in the porous body constituting second layer  32  hardly contain first developing solvent  70 , after the multiple components included in sample  60  are developed by first developing solvent  70 . Therefore, it is unnecessary to dry second layer  32  after the multiple components included in the sample are developed in first development direction X. TLC plate  100  can be brought into contact with second developing solvent  71  with first layer  31  being impregnated with first developing solvent  70 . In other words, it is unnecessary to heat TLC plate  100  to a temperature higher than room temperature or leave TLC plate  100  in an atmosphere with a pressure lower than atmospheric pressure, during a period from when TLC plate  100  is lifted up from first developing solvent  70  till TCL plate  100  is brought into contact with second developing solvent  71 . According to TLC plate  100 , the multiple components can be developed in second development direction Y just after the development of the multiple components in first development direction X. Thus, the multiple components can be separated from each other more easily and more quickly. Note that, with the sample analysis method in the present exemplary embodiment, separation layer  20  may be dried before TLC plate  100  is brought into contact with second developing solvent  71 . 
     A method for detecting positions of multiple components is not particularly limited, and any known methods can be employed. For example, when first layer  31  and second layer  32  contain a fluorescence indicator, separation layer  20  may be irradiated with ultraviolet ray to detect the positions of multiple components. In such a case, each of the multiple components can be a compound that absorbs ultraviolet ray. The analyzing device may have a mechanism for emitting ultraviolet ray. The positions of the multiple components may be detected by depositing a coloring reagent onto separation layer  20 . In such a case, TLC plate  100  may be heated as necessary. Any known coloring reagent can be used. Examples of the coloring reagent include anisaldehyde, phosphomolybdic acid, iodine, ninhydrin, chameleon solution, 2,4-dinitrophenylhydrazine, manganese chloride, and bromocresol green. 
     Under the same condition, the positions of the multiple components after sample  60  is developed are determined for each component. Therefore, with the sample analysis method according to the present exemplary embodiment, each of the separated multiple components can be identified. For example, a component having a known structure is developed on TLC plate  100  under the condition same as the condition for developing sample  60 . Data in which the position of the component after the development and the structure of the component are associated with each other is acquired. This data may be stored in a memory of the analyzing device in advance. Through comparison with the data, each of the multiple components can be identified based on the position of each component after sample  60  is developed. 
     Second Exemplary Embodiment 
     As shown in  FIGS. 3A and 3B , TLC plate  200  according to the present second exemplary embodiment includes separation layer  21  having first layer  31 , second layer  32 , and functional layer  30 . A structure of TLC plate  200  is the same as the structure of TLC plate  100  in the first exemplary embodiment except for functional layer  30 . Therefore, constituent elements which are common between TLC plate  100  in the first exemplary embodiment and TLC plate  200  in the present exemplary embodiment are denoted by the same reference marks and may not be described in detail below. That is, the descriptions regarding the following exemplary embodiments are mutually applicable, in so far as they are technically consistent with one another. In addition, the respective exemplary embodiments may be combined with one another, in so far as they are technically consistent with one another. 
     Functional layer  30  has a rectangular shape in a plan view. Functional layer  30  extends in second development direction Y. Functional layer  30  is in contact with first layer  31 . When separation layer  21  is viewed in a plan view, one side (long side) of first layer  31  is in contact with one side of functional layer  30 . The length of one side of functional layer  30  is equal to the length of one side (long side) of first layer  31 . First layer  31  and functional layer  30  constitute boundary face  41 . Boundary face  41  extends in first development direction X. Functional layer  30  extends from an end face of substrate  10  in second development direction Y to boundary face  41 . Functional layer  30 , first layer  31 , and second layer  32  are arrayed in this order in second development direction Y. 
     In the present exemplary embodiment, first layer  31 , second layer  32 , and functional layer  30  are disposed on substrate  10 . In other words, first layer  31 , second layer  32 , and functional layer  30  are in contact with substrate  10 . A lateral surface of first layer  31  and a lateral surface of second layer  32  are in contact with each other. A lateral surface of first layer  31  and a lateral surface of functional layer  30  are in contact with each other. When the multiple components are developed in second development direction Y, the developing solvent can easily move from functional layer  30  to first layer  31  through boundary face  41 . 
     Functional layer  30  includes a hydrophobic porous body. The hydrophobic porous body may be the same as any of those described as examples of the porous body constituting second layer  32 . An average pore diameter of functional layer  30  may be within a range from 0.01 μm to 100 μm. When the hydrophobic porous body is an aggregate of inorganic particles modified with a hydrophobic functional group, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. Functional layer  30  may further include any of the additives mentioned above. 
     A composition of functional layer  30  may be the same as or different from a composition of second layer  32 . A structure of functional layer  30  may be the same as or different from a structure of second layer  32 . “The structure of functional layer  30  being different from the structure of second layer  32 ” means that at least one factor selected from among an average pore diameter of the porous body constituting functional layer  30 , a void ratio of the porous body, and an average particle diameter of the material of the porous body is different from that of the porous body constituting second layer  32 , for example. 
     Length L 6  of functional layer  30  of TLC plate  200  in second development direction Y is not particularly limited. Length L 6  is determined according to the porous body constituting functional layer  30 , a size of a container for housing TLC plate  200 , and the like. Length L 6  is 5 mm to 50 mm, for example. 
     As a method for forming functional layer  30  on substrate  10 , the methods described above as examples of the method for forming second layer  32  on substrate  10  in the first exemplary embodiment can be used, for example. 
     Each of second layer  32  and functional layer  30  of TLC plate  200  includes a hydrophobic porous body. Therefore, first developing solvent  70  is difficult to penetrate into each of second layer  32  and functional layer  30 . That is, pores in the porous bodies constituting second layer  32  and functional layer  30  hardly contain first developing solvent  70 , after the multiple components included in sample  60  are developed by first developing solvent  70 . Therefore, it is unnecessary to dry second layer  32  and functional layer  30  after the multiple components included in the sample are developed in first development direction X. 
     According to TLC plate  200 , the multiple components can move straight in second development direction Y in second layer  32 . Specifically, when the multiple components are developed in second development direction Y, an end of functional layer  30  in second development direction Y is brought into contact with second developing solvent  71 . In this case, a gradient may occur in a movement distance of second developing solvent  71  in second development direction Y. When the multiple components are developed with the gradient occurring in the movement distance, the multiple components may move in second layer  32  in a direction different from second development direction Y. However, when the gradient occurs in the movement distance, a portion of second developing solvent  71  moves in first development direction X as well as in second development direction Y. Therefore, the gradient in the movement distance of second developing solvent  71  is reduced, as second developing solvent  71  moves in functional layer  30 . In TLC plate  200 , when second developing solvent  71  moves from functional layer  30  to first layer  31 , the gradient in the movement distance of second developing solvent  71  in second development direction Y is reduced. Thus, the multiple components can move straight in second development direction Y in second layer  32 . 
     Modification of Second Exemplary Embodiment 
     As shown in  FIG. 3C , first layer  31  may be disposed on second layer  32  and functional layer  30 . In TLC plate  210 , second layer  32  and functional layer  30  are disposed on substrate  10 . Second layer  32  is not in contact with functional layer  30 . Space  50  is formed between second layer  32  and functional layer  30 . First layer  31  is in contact with second layer  32  and functional layer  30 . A lower surface of first layer  31  and an upper surface of second layer  32  constitute boundary face  42 . The lower surface of first layer  31  and an upper surface of functional layer  30  constitute boundary face  43 . Boundary faces  42  and  43  extend in first development direction X. When the sample is developed in second development direction Y, second developing solvent  71  moves from functional layer  30  to first layer  31  through boundary face  43 . Second developing solvent  71  moves from first layer  31  to second layer  32  through boundary face  42 . Due to space  50 , second developing solvent  71  does not directly move to second layer  32  from functional layer  30 . Therefore, the multiple components located in first layer  31  can easily move to second layer  32 . 
     TLC plate  210  can be manufactured in such a way that second layer  32  and functional layer  30  are formed on substrate  10 , and then, first layer  31  is formed on second layer  32  and functional layer  30 . As a method for forming first layer  31  on second layer  32  and functional layer  30 , the methods described above as examples of the method for forming first layer  31  on substrate  10  in the first exemplary embodiment can be used, for example. In TLC plate  210 , first layer  31  is formed after the formation of second layer  32  and functional layer  30 , whereby separation layer  21  can be easily manufactured. 
     Another Modification of Second Exemplary Embodiment 
     As shown in  FIG. 3D , second layer  32  may be in contact with functional layer  30 . In TLC plate  220 , a lateral surface of second layer  32  and a lateral surface of functional layer  30  constitute boundary face  44 . First layer  31  is disposed on second layer  32  and functional layer  30 . A lower surface of first layer  31  and upper surfaces of second layer  32  and functional layer  30  constitute boundary face  45 . Boundary faces  44  and  45  extend in first development direction X. When the sample is developed in second development direction Y, the developing solvent moves from functional layer  30  to second layer  32  through boundary face  44 . 
     TLC plate  220  is manufactured in the same manner as TLC plate  210 . In TLC plate  220 , first layer  31  is formed after the formation of second layer  32  and functional layer  30 , whereby separation layer  21  can be easily manufactured. 
     In TLC plate  220 , first developing solvent  70  is difficult to penetrate into each of second layer  32  and functional layer  30 . Therefore, when being developed by first developing solvent  70 , the multiple components included in sample  60  are held in first layer  31 . Then, TLC plate  220  is brought into contact with second developing solvent  71 . At that time, second layer  32  and functional layer  30  are both in contact with second developing solvent  71 . Therefore, the porous bodies constituting second layer  32  and functional layer  30  get wet with second developing solvent  71 . In this case, the multiple components held in first layer  31  tend to move to second layer  32  or functional layer  30  through boundary face  45 . That is, when the porous bodies constituting second layer  32  and functional layer  30  get wet, the multiple components tend to move in the thickness direction of separation layer  21 . This tendency is significant when alcohol is used as the organic solvent contained in second developing solvent  71 . Due to the movement of the multiple components to second layer  32  or functional layer  30 , the multiple components can be developed in second development direction Y. 
     Third Exemplary Embodiment 
     As shown in  FIGS. 4A and 4B , in TLC plate  300  according to the present third exemplary embodiment, first layer  31  is disposed on second layer  32 . Second layer  32  is disposed on substrate  10 . In other words, only second layer  32  is in contact with substrate  10 . Second layer  32  extends from one of a pair of end faces of substrate  10  to the other in second development direction Y. A lower surface of first layer  31  and an upper surface of second layer  32  are in contact with each other. First layer  31  and second layer  32  constitute boundary face  46 . Boundary face  46  extends in first development direction X. First layer  31  is located between one end  32   a  and other end  32   b  of second layer  32  in second development direction Y. 
     A distance from one end  32   a  of second layer  32  to first layer  31  in second development direction Y is equal to a value that can be assumed by length L 6  of functional layer  30  in TLC plate  200 . A distance from first layer  31  to other end  32   b  of second layer  32  in second development direction Y is equal to a value that can be assumed by length L 3  of second layer  32  in TLC plate  100 . 
     TLC plate  300  can be manufactured in such a way that second layer  32  is formed on substrate  10 , and then, first layer  31  is formed on second layer  32 . As a method for forming second layer  32  on substrate  10  and a method for forming first layer  31  on second layer  32 , the methods exemplified in the first exemplary embodiment can be used, for example. In TLC plate  300 , first layer  31  is formed after the formation of second layer  32 , whereby separation layer  22  can be easily manufactured. 
     In TLC plate  300 , first developing solvent  70  is difficult to penetrate into second layer  32 . Therefore, when being developed by first developing solvent  70 , the multiple components included in sample  60  are held in first layer  31 . Then, TLC plate  300  is brought into contact with second developing solvent  71 . At that time, second layer  32  is in contact with second developing solvent  71 . Thus, the porous body constituting second layer  32  gets wet with second developing solvent  71 . In this case, the multiple components held in first layer  31  tend to move to second layer  32  through boundary face  46 . That is, when the porous body constituting second layer  32  gets wet, the multiple components tend to move in the thickness direction of separation layer  22 . 
     This tendency is significant when alcohol is used as the organic solvent contained in second developing solvent  71 . Due to the movement of the multiple components to second layer  32 , the multiple components can be developed in second development direction Y. 
     According to TLC plate  300 , the multiple components can move straight in second development direction Y in second layer  32 . Specifically, when the multiple components are developed in second development direction Y, second developing solvent  71  is brought into contact with one end  32   a  of second layer  32 . In this case, a gradient may occur in a movement distance of second developing solvent  71  in second development direction Y. When the multiple components are developed with the gradient occurring in the movement distance, the multiple components may move in second layer  32  in a direction different from second development direction Y. However, when the gradient occurs in the movement distance, a portion of second developing solvent  71  moves in first development direction X as well as in second development direction Y. Therefore, the gradient in the movement distance of second developing solvent  71  is reduced, as second developing solvent  71  moves in second layer  32 . In TLC plate  300 , when second developing solvent  71  moves from one end  32   a  of second layer  32  to first layer  31 , the gradient in the movement distance of second developing solvent  71  in second development direction Y is reduced. Thus, the multiple components can move straight in second development direction Y in second layer  32 . 
     Fourth Exemplary Embodiment 
     The TLC plate may further include a pair of electrodes. In  FIGS. 5A and 5B , TLC plate  400  has a pair of electrodes  55 . The pair of electrodes  55  is disposed at both ends of first layer  31  in first development direction X. The pair of electrodes  55  is disposed on first layer  31 . If a voltage is applied to the pair of electrodes  55  with first layer  31  being impregnated with first developing solvent  70 , current flows through first layer  31 . A structure of TLC plate  400  is the same as the structure of TLC plate  300  in the third exemplary embodiment except for the pair of electrodes  55 . Note that an average pore diameter of the porous material constituting first layer  31  of TLC plate  400  may be within a range from 0.1 μm to 100 μm. With this configuration, the multiple components included in the sample can be easily electrophoresed in first layer  31 . 
     The pair of electrodes  55  is not particularly limited, as long as they can apply a voltage. The pair of electrodes  55  may be formed from at least one metal selected from the group consisting of platinum, gold, copper, and aluminum, for example. 
     Next, a sample analysis method using TLC plate  400  will be described. 
     First, sample  60  is placed on first layer  31  of separation layer  22  of TLC plate  400 , as shown in  FIG. 6A . When sample  60  is placed on first layer  31 , sample  60  penetrates into first layer  31 , so that circular spot  61  is formed. Sample  60  is an aqueous solution containing a plurality of proteins, for example. In first layer  31 , a position where sample  60  is to be placed is not particularly limited. Sample  60  may be placed on a middle point of first layer  31  in first development direction X. In this case, the multiple components included in sample  60  can be quickly separated from each other by electrophoresis of the multiple components. First layer  31  is impregnated with first developing solvent  70  in advance. First developing solvent  70  is typically the same as that used in the first exemplary embodiment. 
     Next, a voltage is applied to electrodes  55  on TLC plate  400  as shown in  FIG. 6B . The multiple components included in sample  60  are electrophoresed in first development direction X. The voltage can be applied by power source  80 . Power source  80  is an AC-to-DC converter, a power generating device, or a battery, for example. The electrophoresis may be conducted inside an analyzing device. The multiple components are separated from each other in first layer  31  based on isoelectric point or molecular weight of each component. Due to the electrophoresis of the multiple components in first development direction X, spots  66 ,  67 ,  68 , and  69  are newly generated. 
     Then, as shown in  FIG. 6C , TLC plate  400  is placed in container  76  with end  32   a  of second layer  32  in second development direction Y being directed downward. Container  76  contains second developing solvent  71 . Container  76  and second developing solvent  71  are typically the same as those used in the first exemplary embodiment. 
     When TLC plate  400  is placed in container  76 , end  32   a  of second layer  32  is in contact with second developing solvent  71 . At that time, first layer  31  is impregnated with first developing solvent  70 . The liquid level of second developing solvent  71  is set to prevent direct contact between second developing solvent  71  and spots  66 ,  67 ,  68 , and  69 . The porous body constituting second layer  32  gets wet with second developing solvent  71  by second developing solvent  71 . Thus, the multiple components move from first layer  31  to second layer  32 . Due to capillary force, second developing solvent  71  moves in second development direction Y from end  32   a  of second layer  32 . When second developing solvent  71  is brought into contact with the multiple components located in spots  66 ,  67 ,  68 , and  69 , the multiple components are dissolved into second developing solvent  71 . The multiple components dissolved in second developing solvent  71  move in second development direction Y along with second developing solvent  71 . The multiple components which are not separated from each other in first layer  31  are separated from each other in second layer  32 . 
     Second layer  32  of TLC plate  400  includes a hydrophobic porous body. First developing solvent  70  hardly penetrates into second layer  32 . That is, pores in the porous body constituting second layer  32  hardly contain first developing solvent  70  after the electrophoresis of the multiple components included in sample  60 . Therefore, it is unnecessary to dry second layer  32  after the multiple components included in the sample are developed in first development direction X. TLC plate  400  can be brought into contact with second developing solvent  71  with first layer  31  being impregnated with first developing solvent  70 . In other words, it is unnecessary to heat TLC plate  400  to a temperature higher than room temperature or leave TLC plate  400  in an atmosphere with a pressure lower than atmospheric pressure, during a period from when the voltage applied to electrodes  55  is removed till TLC plate  400  is brought into contact with second developing solvent  71 . According to TLC plate  400 , the multiple components can be developed in second development direction Y just after the development of the multiple components in first development direction X. Thus, the multiple components can be separated from each other more easily and more quickly. Note that, with the sample analysis method in the present exemplary embodiment, separation layer  22  may be dried before TLC plate  400  is brought into contact with second developing solvent  71 . 
     With the sample analysis method according to the present exemplary embodiment, it is unnecessary to change the orientation of TLC plate  400 . Therefore, the analyzing device used for the sample analysis method according to the present exemplary embodiment does not need a mechanism for changing the orientation of the TLC plate. 
     As a method for detecting the positions of the multiple components and a method for identifying each of the multiple components in the sample analysis method according to the present exemplary embodiment, the methods described in the first exemplary embodiment can be used. 
     Fifth Exemplary Embodiment 
     As shown in  FIGS. 7A and 7B , TLC plate  500  according to the present fifth exemplary embodiment is obtained by further providing third layer  33  to the configuration of TLC plate  100  in the first exemplary embodiment. Third layer  33  induces an interaction different from the interaction induced by second layer  32 , with respect to multiple components included in a sample. Therefore, the multiple components which are not separated from each other in second layer  32  are separated from each other in third layer  33 . 
     Third layer  33  has a rectangular shape in a plan view. Third layer  33  extends in second development direction Y. Third layer  33  is in contact with second layer  32 . When separation layer  23  is viewed in a plan view, one side of second layer  32  is in contact with one side of third layer  33 . The length of one side of third layer  33  is equal to the length of one side of second layer  32 . Third layer  33  and second layer  32  constitute boundary face  47 . Boundary face  47  extends in first development direction X. Third layer  33  extends from boundary face  47  to an end face of substrate  10  in second development direction Y. First layer  31 , second layer  32 , and third layer  33  are arrayed in this order in second development direction Y. 
     In the present exemplary embodiment, first layer  31 , second layer  32 , and third layer  33  are disposed on substrate  10 . In other words, first layer  31 , second layer  32 , and third layer  33  are in contact with substrate  10 . A lateral surface of first layer  31  and a lateral surface of second layer  32  are in contact with each other. A lateral surface of second layer  32  and a lateral surface of third layer  33  are in contact with each other. When the multiple components are developed in second development direction Y, a developing solvent can easily move from second layer  32  to third layer  33  through boundary face  47 . 
     Third layer  33  includes a porous body. The porous body constituting third layer  33  may be the same as any of those described as examples of the porous body constituting first layer  31  or second layer  32 . When the multiple components included in the sample are developed in first development direction X by electrophoresis, third layer  33  may includes a hydrophilic porous body. An average pore diameter of third layer  33  may be within a range from 0.01 μm to 100 μm. When the porous body is an aggregate of inorganic particles, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. Third layer  33  may further include any of the additives mentioned above. 
     TLC plate  500  satisfies at least one requirement selected from among a requirement in which a composition of third layer  33  is different from a composition of second layer  32  and a requirement in which a structure of third layer  33  is different from a structure of second layer  32 . Thus, third layer  33  induces an interaction different from the interaction induced by second layer  32 , with respect to the multiple components included in the sample. “The structure of third layer  33  being different from the structure of second layer  32 ” means that at least one factor selected from among an average pore diameter of the porous body constituting third layer  33 , a void ratio of the porous body, and an average particle diameter of the material of the porous body is different from that of the porous body constituting second layer  32 , for example. The multiple components which are not separated from each other in second layer  32  are separated from each other in third layer  33 . 
     Length L 7  of second layer  32  in second development direction Y is not particularly limited. Length L 7  is determined according to the porous body constituting second layer  32 , a size of a container for housing TLC plate  500 , and the like. 
     Length L 8  of third layer  33  in second development direction Y is not particularly limited. Length L 8  is determined according to the porous body constituting third layer  33 , a size of a container for housing TLC plate  500 , and the like. 
     As a method for forming third layer  33  on substrate  10 , the methods described above as examples of the method for forming first layer  31  on substrate  10  and the method for forming second layer  32  on substrate  10  in the first exemplary embodiment can be used, for example. 
     TLC plate  500  may further include functional layer  30  provided to TLC plate  200  in the second exemplary embodiment. In this configuration, functional layer  30 , first layer  31 , second layer  32 , and third layer  33  are arrayed in this order in second development direction Y. 
     Sixth Exemplary Embodiment 
     As shown in  FIGS. 8A and 8B , thin layer chromatography plate  600  (hereinafter referred to as “TLC plate  600 ”) according to the present first exemplary embodiment has substrate  10  and separation layer  20 . Substrate  10  has a plate shape, for example. Substrate  10  has, for example, a rectangular shape in a plan view. Separation layer  20  is disposed on substrate  10 . Separation layer  20  covers the surface of substrate  10 . Substrate  10  has two pairs of end faces facing each other. In the present exemplary embodiment, first development direction X extends from one end face of one of two pairs of end faces of substrate  10  to the other end face. Multiple components included in the sample are developed in first development direction X in a first stage. Second development direction Y extends from one end face of the other of two pairs of end faces of substrate  10  to the other end face. Multiple components included in the sample are developed in second development direction Y in a second stage. Second development direction Y is orthogonal to first development direction X. 
     Separation layer  20  separates the multiple components included in the sample from each other. Separation layer  20  includes first layer  31 , second layer  32 , and third layer  33 . First layer  31  is a band-shaped layer. First layer  31  has a rectangular band shape in a plan view. First layer  31  extends in first development direction X. First layer  31  extends from one of a pair of end faces of substrate  10  to the other in first development direction X. Note that first layer  31  may not extend to the other end face of substrate  10 . 
     Second layer  32  has a rectangular shape in a plan view. Second layer  32  extends in second development direction Y. Second layer  32  is in contact with first layer  31 . When separation layer  20  is viewed in a plan view, one side (long side) of first layer  31  is in contact with one side of second layer  32 . 
     The length of one side (long side) of first layer  31  is equal to the length of one side of second layer  32 . First layer  31  and second layer  32  constitute boundary face  40 . Boundary face  40  extends in first development direction X. Second layer  32  extends to third layer  33  from boundary face  40 . 
     Third layer  33  has a rectangular shape in a plan view. Third layer  33  extends in second development direction Y. Third layer  33  is in contact with second layer  32 . When separation layer  20  is viewed in a plan view, one side of second layer  32  is in contact with one side of third layer  33 . The length of one side of third layer  33  is equal to the length of one side of second layer  32 . Third layer  33  and second layer  32  constitute boundary face  41 . Boundary face  41  extends in first development direction X. Third layer  33  extends from boundary face  41  to an end face of substrate  10  in second development direction Y. Note that third layer  33  may not extend to the end face of substrate  10 . First layer  31 , second layer  32 , and third layer  33  are arrayed in this order in second development direction Y. 
     In the present exemplary embodiment, first layer  31 , second layer  32 , and third layer  33  are disposed on substrate  10 . In other words, first layer  31 , second layer  32 , and third layer  33  are in contact with substrate  10 . A lateral surface of first layer  31  and a lateral surface of second layer  32  are in contact with each other. A lateral surface of second layer  32  and a lateral surface of third layer  33  are in contact with each other. When the multiple components are developed in second development direction Y, a developing solvent can easily move from first layer  31  to second layer  32  through boundary face  40 . The developing solvent can easily move from second layer  32  to third layer  33  through boundary face  41 . 
     The material of substrate  10  is not particularly limited, as long as it can maintain the shape of TLC plate  600  without being eluted in the developing solvent. The material of substrate  10  is glass, resin, metal, or paper, for example. Substrate  10  is typically a glass plate or an aluminum film. 
     First layer  31  includes a hydrophilic porous body. In the present specification, “being hydrophilic” means that, when water is brought into contact with the porous body, water can move in the porous body at a rate of 5 mm/min or higher due to capillary force, for example. When water is selected as the developing solvent, first layer  31  can carry water from one end to the other end of first layer  31  in first development direction X due to capillary force. The hydrophilic porous body is not particularly limited. The material of the hydrophilic porous body includes at least one selected from the group consisting of a fiber material, and an inorganic material and a polymer material which are hydrophilic, for example. 
     The fiber material includes at least one selected from the group consisting of a plant fiber, an animal fiber, a recycled fiber, a synthetic fiber, and a glass fiber, for example. The plant fiber includes cellulose, for example. The synthetic fiber includes cellulose acetate, for example. 
     The hydrophilic polymer material includes at least one selected from the group consisting of agarose, dextran, and mannan, for example. The inorganic material includes at least one selected from the group consisting of alumina, silicon dioxide, and zirconia, for example. 
     The hydrophilic porous body is filter paper, for example. The hydrophilic porous body is an aggregate of at least one kind of inorganic particles selected from the group consisting of alumina particles, silica gel particles, silicon pillar, zeolite particles, diatom earth, and zirconia particles, for example. 
     An average pore diameter of first layer  31  may be within a range from 0.01 μm to 100 μm. When the hydrophilic porous body is an aggregate of inorganic particles, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. The “average pore diameter” can be measured with the following method. Specifically, the surface or cross-section of first layer  31  is observed with an electron microscope (for example, an electron scanning microscope). Pore diameters of a plurality of observed pores (for example, random 50 pores) are measured. The average pore diameter is determined based on the average value calculated using the measured values. The diameter of a circle having an area equal to the area of the pore observed with the electron microscope can be regarded as the pore diameter. The “average particle diameter” can be measured with the following method. Specifically, the surface or cross-section of first layer  31  is observed with an electron microscope, and diameters of random number (for example, 50) of particles constituting first layer  31  are measured. The average particle diameter is determined based on the average value calculated using the obtained measured values. The diameter of a circle having an area equal to the area of the particle observed with the electron microscope can be regarded as the particle diameter. 
     First layer  31  may further include an additive. Examples of the additive include a fluorescence indicator, a binder, and a metal oxide. 
     Examples of the fluorescence indicator include magnesium tungstate and zinc silicate containing manganese. When first layer  31  includes the fluorescence indicator, positions of the multiple components can be detected by irradiating first layer  31  with ultraviolet ray. 
     The binder includes at least one selected from the group consisting of an inorganic binder, an organic fiber, a thickener, and an organic binder, for example. Examples of the inorganic binder include plaster and colloidal silica. Examples of the organic fiber include microfibrillar cellulose. Examples of the thickener include hydroxyethyl cellulose and carboxymethyl cellulose. Examples of the organic binder include polyvinyl alcohol and polyacrylic acid. When first layer  31  includes the binder, adhesiveness between substrate  10  and first layer  31  is improved. When the porous body of first layer  31  is an aggregate of inorganic particles, durability of the aggregate of inorganic particles is improved due to the binder. 
     The metal oxide includes at least one selected from the group consisting of titanium oxide, aluminum oxide, tin oxide, zinc oxide, tungsten oxide, manganese oxide, nickel oxide, copper oxide, and magnesium oxide, for example. The metal oxide may be charged when getting wet with the developing solvent. Therefore, when first layer  31  includes the metal oxide, the interaction between the multiple components included in the sample and first layer  31  varies. Thus, the multiple components may be easily separated from each other in first layer  31 . 
     The above-mentioned additives may be mixed into the porous body constituting first layer  31 . When the additive is a binder or a metal oxide, the additive may cover the surface of the porous body. The additive may coat the surfaces of inorganic particles constituting the porous body. 
     Second layer  32  includes a hydrophobic porous body. In the present specification, “being hydrophobic” means that, when water is brought into contact with the porous body, water can move in the porous body at a rate lower than  5  mm/min due to capillary force, or water does not move inside the porous body due to capillary force, for example. When water moves inside the hydrophobic porous body, the movement rate of water may be lower than 1 mm/min. When second layer  32  is brought into contact with water, water hardly penetrates into second layer  32 . When the developing solvent contains an organic solvent, second layer  32  can carry the developing solvent from one end to the other end of second layer  32  in second development direction Y due to capillary force. The hydrophobic porous body is not particularly limited. 
     The material of the hydrophobic porous body is, for example, a hydrophobic polymer material. The hydrophobic polymer material includes at least one selected from the group consisting of fluororesin, polystyrene, polyethylene, and polypropylene, for example. 
     The hydrophobic porous body is, for example, a porous body of a hydrophobic polymer material or an aggregate of hydrophobic polymer material particles. The hydrophobic porous body is an aggregate of inorganic particles modified with a hydrophobic functional group, for example. The hydrophobic functional group includes a functional group having a hydrocarbon group at the end, for example. The hydrocarbon group includes at least one selected from the group consisting of an octadecyl group, an octyl group, a t-butyl group, a trimethylsilyl group, and a phenyl group, for example. The inorganic particles include at least one kind selected from the group consisting of alumina particles, silica gel particles, silicon pillar, zeolite particles, diatom earth, and zirconia particles, for example. The hydrophobic porous body is typically an aggregate of silica gel particles modified with a hydrophobic functional group. Whether the inorganic particles are modified with a hydrophobic functional group can be confirmed by conducting an element analysis on the cross-section of second layer  32 , for example. The element analysis can be conducted by X-ray photoelectron spectroscopy (XPS) or energy dispersive X-ray spectroscopy (EDX), for example. 
     An average pore diameter of second layer  32  may be within a range from 0.01 μm to 100 μm. When the hydrophobic porous body is an aggregate of inorganic particles modified with a hydrophobic functional group, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. Second layer  32  may further include any of the above-mentioned additives. 
     Third layer  33  includes a porous body. The porous body constituting third layer  33  may be the same as any of those described as examples of the porous body constituting first layer  31  or second layer  32 . When the multiple components included in the sample are developed in first development direction X by electrophoresis, third layer  33  may include a hydrophilic porous body. An average pore diameter of third layer  33  may be within a range from 0.01 μm to 100 μm. When the porous body is an aggregate of inorganic particles, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. Third layer  33  may further include any of the additives mentioned above. 
     TLC plate  600  satisfies at least one requirement selected from among a requirement in which a composition of third layer  33  is different from a composition of second layer  32  and a requirement in which a structure of third layer  33  is different from a structure of second layer  32 . Thus, third layer  33  induces an interaction different from the interaction induced by second layer  32 , with respect to the multiple components included in the sample. “The structure of third layer  33  being different from the structure of second layer  32 ” means that at least one factor selected from among an average pore diameter of the porous body constituting third layer  33 , a void ratio of the porous body, and an average particle diameter of the material of the porous body is different from that of the porous body constituting second layer  32 , for example. 
     Length L 11  of first layer  31  in first development direction X is not particularly limited. Length L 11  is determined according to the porous body constituting first layer  31 , a size of a container for housing TLC plate  600 , and the like. Length L 11  is 10 mm to 100 mm, for example. The length of second layer  32  and the length of substrate  10  in first development direction X are typically equal to length L 11 . 
     Length L 12  of first layer  31  in second development direction Y is not particularly limited. Length L 12  is determined according to an amount of the sample to be placed on first layer  31 , for example. As length L 12  is smaller, the multiple components are more easily separated from each other when the multiple components are developed in second development direction Y. Length L 12  is 0.5 mm to 10 mm, for example. 
     Length L 13  of second layer  32  in second development direction Y is not particularly limited. Length L 13  is determined according to the porous body constituting second layer  32 , a size of a container for housing TLC plate  600 , and the like. 
     Length L 14  of third layer  33  in second development direction Y is not particularly limited. Length L 14  is determined according to the porous body constituting third layer  33 , a size of a container for housing TLC plate  600 , and the like. The length of substrate  10  in second development direction Y is typically equal to the total of length L 12 , length L 13 , and length L 14 . 
     Thickness L 15  of first layer  31  is not particularly limited. Thickness L 15  is determined according to the porous body constituting first layer  31 . Thickness L 15  is 0.05 mm to 1 mm, for example. The thickness of second layer  32  and the thickness of third layer  33  are typically equal to thickness L 15  of first layer  31 . 
     Thickness L 16  of substrate  10  is not particularly limited as long as the shape of TLC plate  600  can be maintained. Thickness L 16  is 0.1 mm to 5 mm, for example. 
     Next, a manufacturing method of TLC plate  100  will be described. 
     First, a first dispersion liquid containing inorganic particles is prepared. The first dispersion liquid can be obtained by dispersing inorganic particles into a coating solvent. 
     The coating solvent includes at least one selected from the group consisting of water and an organic solvent, for example. The organic solvent includes at least one selected from the group consisting of alcohol, ketone, ether, nitrile, sulfoxide, sulfone, ester, carboxylic acid, amide, hydrocarbon, aromatic hydrocarbon, and halogen-containing compound, for example. Examples of alcohol include methanol, ethanol, and isopropyl alcohol. Examples of ketone include acetone and ethyl methyl ketone. Examples of ether include tetrahydrofuran and dioxane. Examples of nitrile include acetonitrile. Examples of sulfoxide include dimethyl sulfoxide. Examples of sulfone include sulfolane. Examples of ester include ethyl acetate. Examples of carboxylic acid includes formic acid and acetic acid. Examples of amide include dimethylformamide. Examples of hydrocarbon include pentane and hexane. Examples of aromatic hydrocarbon include benzene, toluene, and xylene. Examples of halogen-containing compound include methylene chloride, chloroform, bromoform, chlorobenzene, and bromobenzene. 
     The first dispersion liquid is applied on a part of the surface of substrate  10  to form a coating film. The coating film is dried, whereby first layer  31  is formed on substrate  10 . When the hydrophilic porous body is filter paper, first layer  31  is formed on substrate  10  by bonding the hydrophilic porous body to a part of the surface of substrate  10  under pressure. 
     Next, a second dispersion liquid containing inorganic particles modified with a hydrophobic functional group is prepared. The second dispersion liquid can be obtained by dispersing inorganic particles modified with a hydrophobic functional group into a coating solvent. The materials mentioned above can be used for the coating solvent. 
     The second dispersion liquid is applied on a part of the surface of substrate  10  to form a coating film. The coating film is dried, whereby second layer  32  is formed on substrate  10 . When the hydrophobic porous body is a porous body of a hydrophobic polymer material, second layer  32  is formed on substrate  10  by bonding the hydrophobic porous body to a part of the surface of substrate  10  under pressure. 
     The second dispersion liquid may contain inorganic particles not modified with a hydrophobic functional group, in place of inorganic particles modified with a hydrophobic functional group. In such a case, second layer  32  is formed in the manner described below. The second dispersion liquid is applied on substrate  10  to form a coating film. The coating film is dried, whereby a untreated layer of second layer  32  is formed. A silane coupling agent having a hydrophobic functional group is applied on the untreated layer. The silane coupling agent is reacted with the inorganic particles included in the untreated layer. Thus, second layer  32  is formed on substrate  10 . The silane coupling agent may be applied on a coating film, not on the untreated layer. 
     The silane coupling agent is not particularly limited. The silane coupling agent may include at least one selected from the group consisting of dimethyloctadecylchlorosilane, dimethyloctylchlorosilane, t-butyldimethylchlorosilane, trimethylchlorosilane, and phenyldimethylchlorosilane. 
     First layer  31  and second layer  32  may be formed by the following method. The first dispersion liquid is applied on the entire surface of substrate  10  to form a coating film The coating film is dried, whereby a untreated layer of second layer  32  and first layer  31  are formed on substrate  10 . A silane coupling agent having a hydrophobic functional group is applied on the untreated layer of second layer  32 . The silane coupling agent is reacted with the inorganic particles. Thus, first layer  31  and second layer  32  are formed on substrate  10 . 
     Next, third layer  33  is formed on substrate  10 . As a method for forming third layer  33  on substrate  10 , the methods described above as examples of the method for forming first layer  31  on substrate  10  and the method for forming second layer  32  on substrate  10  can be used, for example. 
     The order of formation of first layer  31 , second layer  32 , and third layer  33  on substrate  10  is not particularly limited. First layer  31  and second layer  32  may be respectively formed on substrate  10  after third layer  33  is formed on substrate  10 . 
     Next, a sample analysis method using TLC plate  600  will be described. 
     First, sample  60  is placed on first layer  31  of separation layer  20  of TLC plate  600 , as shown in  FIG. 9A . When sample  60  is placed on first layer  31 , sample  60  penetrates into first layer  31 , so that circular spot  61  is formed. Sample  60  is an aqueous solution containing a plurality of proteins, for example. The content of the plurality of proteins in sample  60  is from 0.01 wt. % to 1 wt. %, for example. The volume of sample  60  placed on first layer  31  is 0.1 μL to 2 μL, for example. The position where sample  60  is to be placed on first layer  31  is not particularly limited, as long as sample  60  is not in direct contact with the first developing solvent and the second developing solvent. 
     Then, as shown in  FIG. 9B , TLC plate  600  is placed in container  75  with end  31   a  of first layer  31  in first development direction X being directed downward. Container  75  contains first developing solvent  70 . Container  75  is a glass jar, for example. Container  75  may be installed inside an analyzing device (not shown). 
     First developing solvent  70  is not particularly limited, as long as it does not move to the inside of second layer  32  or third layer  33  when being in contact with the surface of second layer  32  or third layer  33 . In the sample analysis method according to the present exemplary embodiment, third layer  33  can include a hydrophobic porous body. First developing solvent  70  is water or an aqueous solution, for example. A solute of the aqueous solution contains at least one selected from the group consisting of phosphate, citrate, acetate, and borate, for example. The aqueous solution may be a buffer solution such as a phosphate buffer solution, a tris buffer solution, a citrate buffer solution, an acetate buffer solution, or a borate buffer solution. In the present exemplary embodiment, first developing solvent  70  does not contain an organic solvent. However, first developing solvent  70  may contain an organic solvent. When first developing solvent  70  contains an organic solvent, first developing solvent  70  typically contains 80 volume % or more water. 
     When TLC plate  600  is placed in container  75 , end  31   a  of first layer  31  is in contact with first developing solvent  70 . The liquid level of first developing solvent  70  is set to prevent direct contact between first developing solvent  70  and sample  60 . Due to the capillary force, first developing solvent  70  moves in first development direction X from end  31   a  of first layer  31 . When first developing solvent  70  and sample  60  are brought into contact with each other, the multiple components included in sample  60  are dissolved into first developing solvent  70 . The multiple components dissolved in first developing solvent  70  move in first development direction X along with first developing solvent  70 . The multiple components move while repeatedly adsorbing and desorbing to and from the porous body constituting first layer  31 . The frequency of adsorption and desorption varies in each component, and thus, the multiple components are separated from each other in first layer  31 . Due to the development of sample  60  in first development direction X, spots  62 ,  63 ,  64 , and  65  are newly generated. Spots  62 ,  63 ,  64 , and  65  respectively indicate that any of the multiple components included in the sample is located therein. 
     Then, the orientation of TLC plate  600  is changed. The analyzing device may include a mechanism for changing the orientation of TLC plate  600 . As shown in  FIG. 9C , TLC plate  600  is placed in container  76  with end  31   b  of first layer  31  in second development direction Y being directed downward. Container  76  contains second developing solvent  71 . Container  76  is a glass jar, for example. Container  76  may be installed inside the analyzing device. 
     Second developing solvent  71  is not particularly limited, as long as it contains an organic solvent. Second developing solvent  71  contains an organic solvent, so that it can penetrate into second layer  32  and third layer  33 . The materials mentioned above as examples of the coating solvent can be used as the organic solvent. The organic solvent contains at least one selected from the group consisting of methanol, ethanol, isopropyl alcohol, acetonitrile, and acetic acid, for example. When second developing solvent  71  contains carboxylic acid and the sample contains proteins, the frequency of absorption and desorption of proteins to and from the porous bodies constituting second layer  32  and third layer  33  is improved. Second developing solvent  71  may contain the organic solvent in an amount of 20 wt. % or more. Second developing solvent  71  may contain water in addition to the organic solvent. That is, second developing solvent  71  may be a mixed solvent containing the organic solvent and water. When second developing solvent  71  contains water and the sample contains proteins, solubility of the proteins in second developing solvent  71  is improved. In other words, the multiple components included in the sample can be easily dissolved in second developing solvent  71 . Specific examples of second developing solvent  71  include a mixed solvent in which isopropyl alcohol, acetic acid, and water are mixed in a weight ratio of 40:5:55. 
     When TLC plate  600  is placed in container  76 , end  31   b  of first layer  31  is in contact with second developing solvent  71 . At that time, first layer  31  is impregnated with first developing solvent  70 . The liquid level of second developing solvent  71  is set to prevent direct contact between second developing solvent  71  and spots  62 ,  63 ,  64 , and  65 . Second developing solvent  71  penetrates into first layer  31 . Second developing solvent  71  moves from end  31   b  of first layer  31  in second development direction Y due to capillary force, along with first developing solvent  70  penetrating into first layer  31 . When second developing solvent  71  is brought into contact with the multiple components located in spots  62 ,  63 ,  64 , and  65 , the multiple components are dissolved into second developing solvent  71 . The multiple components dissolved in second developing solvent  71  move in second development direction Y along with second developing solvent  71 . The multiple components move while repeatedly adsorbing and desorbing to and from the porous body constituting second layer  32  or third layer  33 . The multiple components which are not separated from each other in first layer  31  are separated from each other in second layer  32 . The multiple components which are not separated from each other in second layer  32  are separated from each other in third layer  33 . 
     Second layer  32  of TLC plate  600  includes a hydrophobic porous body. First developing solvent  70  hardly penetrates into second layer  32 . That is, pores in the porous body constituting second layer  32  hardly contain first developing solvent  70 , after the multiple components included in sample  60  are developed by first developing solvent  70 . Therefore, it is unnecessary to dry second layer  32  after the multiple components included in the sample are developed in first development direction X. TLC plate  100  can be brought into contact with second developing solvent  71  with first layer  31  being impregnated with first developing solvent  70 . In other words, it is unnecessary to heat TLC plate  600  to a temperature higher than room temperature or leave TLC plate  600  in an atmosphere with a pressure lower than atmospheric pressure, during a period from when TLC plate  600  is lifted up from first developing solvent  70  till TLC plate  600  is brought into contact with second developing solvent  71 . According to TLC plate  600 , the multiple components can be developed in second development direction Y just after the development of the multiple components in first development direction X. Thus, the multiple components can be separated from each other more easily and more quickly. Note that, with the sample analysis method in the present exemplary embodiment, separation layer  20  may be dried before TLC plate  600  is brought into contact with second developing solvent  71 . 
     A method for detecting positions of multiple components is not particularly limited, and any known methods can be employed. For example, when each of first layer  31 , second layer  32 , and third layer  33  contains a fluorescence indicator, separation layer  20  may be irradiated with ultraviolet ray to detect the positions of multiple components. In such a case, each of the multiple components can be a compound that absorbs ultraviolet ray. The analyzing device may have a mechanism for emitting ultraviolet ray. The positions of the multiple components may be detected by depositing a coloring reagent onto separation layer  20 . In such a case, TLC plate  600  may be heated as necessary. Any known coloring reagent can be used. Examples of the coloring reagent include anisaldehyde, phosphomolybdic acid, iodine, ninhydrin, chameleon solution, 2,4-dinitrophenylhydrazine, manganese chloride, and bromocresol green. 
     Under the same condition, the positions of the multiple components after sample  60  is developed are determined for each component. Therefore, with the sample analysis method according to the present exemplary embodiment, each of the separated multiple components can be identified. For example, a component having a known structure is developed on TLC plate  600  under the condition same as the condition for developing sample  60 . Data in which the position of the component after the development and the structure of the component are associated with each other is acquired. This data may be stored in a memory of the analyzing device in advance. Through comparison with the data, each of the multiple components can be identified based on the position of each component after sample  60  is developed. 
     Seventh Exemplary Embodiment 
     As shown in  FIGS. 10A and 10B , TLC plate  700  according to the present seventh exemplary embodiment includes separation layer  21  having first layer  31 , second layer  32 , third layer  33 , and functional layer  30 . A structure of TLC plate  700  is the same as the structure of TLC plate  600  in the sixth exemplary embodiment except for functional layer  30 . Therefore, constituent elements which are common between TLC plate  700  in the first exemplary embodiment and TLC plate  700  in the present exemplary embodiment are denoted by the same reference marks and may not be described in detail below. That is, the descriptions regarding the following exemplary embodiments are mutually applicable, in so far as they are technically consistent with one another. In addition, the respective exemplary embodiments may be combined with one another, in so far as they are technically consistent with one another. 
     Functional layer  30  has a rectangular shape in a plan view. Functional layer  30  extends in second development direction Y. Functional layer  30  is in contact with first layer  31 . When separation layer  21  is viewed in a plan view, one side (long side) of first layer  31  is in contact with one side of functional layer  30 . The length of one side of functional layer  30  is equal to the length of one side (long side) of first layer  31 . First layer  31  and functional layer  30  constitute boundary face  42 . Boundary face  42  extends in first development direction X. Functional layer  30  extends from an end face of substrate  10  in second development direction Y to boundary face  42 . Functional layer  30 , first layer  31 , second layer  32 , and third layer  33  are arrayed in this order in second development direction Y. 
     In the present exemplary embodiment, first layer  31 , second layer  32 , third layer  33 , and functional layer  30  are disposed on substrate  10 . In other words, first layer  31 , second layer  32 , third layer  33 , and functional layer  30  are in contact with substrate  10 . A lateral surface of first layer  31  and a lateral surface of second layer  32  are in contact with each other. A lateral surface of second layer  32  and a lateral surface of third layer  33  are in contact with each other. A lateral surface of first layer  31  and a lateral surface of functional layer  30  are in contact with each other. When the multiple components are developed in second development direction Y, a developing solvent can easily move from functional layer  30  to first layer  31  through boundary face  42 . 
     Functional layer  30  includes a hydrophobic porous body. The hydrophobic porous body may be the same as any of those described as examples of the porous body constituting second layer  32 . An average pore diameter of functional layer  30  may be within a range from 0.01 μm to 100 μm. When the hydrophobic porous body is an aggregate of inorganic particles modified with a hydrophobic functional group, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. Functional layer  30  may further include any of the additives mentioned above. 
     A composition of functional layer  30  may be the same as or different from a composition of second layer  32 . A structure of functional layer  30  may be the same as or different from a structure of second layer  32 . “The structure of functional layer  30  being different from the structure of second layer  32 ” means that at least one factor selected from among an average pore diameter of the porous body constituting functional layer  30 , a void ratio of the porous body, and an average particle diameter of the material of the porous body is different from that of the porous body constituting second layer  32 , for example. 
     Length L 17  of functional layer  30  in second development direction Y is not particularly limited in TLC plate  700 . Length L 17  is determined according to the porous body constituting functional layer  30 , a size of a container for housing TLC plate  700 , and the like. Length L 17  is 5 mm to 50 mm, for example. 
     As a method for forming functional layer  30  on substrate  10 , the methods described above as examples of the method for forming second layer  32  on substrate  10  in the sixth exemplary embodiment can be used, for example. 
     Each of second layer  32  and functional layer  30  of TLC plate  700  includes a hydrophobic porous body. Therefore, first developing solvent  70  is difficult to penetrate into each of second layer  32  and functional layer  30 . That is, pores in the porous bodies constituting second layer  32  and functional layer  30  hardly contain first developing solvent  70 , after the multiple components included in sample  60  are developed by first developing solvent  70 . Therefore, it is unnecessary to dry second layer  32  and functional layer  30  after the multiple components included in the sample are developed in first development direction X. 
     According to TLC plate  700 , the multiple components can move straight in second development direction Y in second layer  32  and third layer  33 . Specifically, when the multiple components are developed in second development direction Y, an end of functional layer  30  in second development direction Y is brought into contact with second developing solvent  71 . In this case, a gradient may occur in a movement distance of second developing solvent  71  in second development direction Y. When the multiple components are developed with the gradient occurring in the movement distance, the multiple components may move in second layer  32  and third layer  33  in a direction different from second development direction Y. However, when the gradient occurs in the movement distance, a portion of second developing solvent  71  moves in first development direction X as well as in second development direction Y. Therefore, the gradient in the movement distance of second developing solvent  71  is reduced, as second developing solvent  71  moves in functional layer  30 . In TLC plate  700 , when second developing solvent  71  moves from functional layer  30  to first layer  31 , the gradient in the movement distance of second developing solvent  71  in second development direction Y is reduced. Thus, the multiple components can move straight in second development direction Y in second layer  32  and third layer  33 . 
     Modification of Seventh Exemplary Embodiment 
     As shown in  FIG. 10C , first layer  31  may be disposed on second layer  32  and functional layer  30 . In TLC plate  710 , second layer  32 , third layer  33 , and functional layer  30  are disposed on substrate  10 . Second layer  32  is not in contact with functional layer  30 . Space  50  is formed between second layer  32  and functional layer  30 . First layer  31  is in contact with second layer  32  and functional layer  30 . A lower surface of first layer  31  and an upper surface of second layer  32  constitute boundary face  43 . The lower surface of first layer  31  and an upper surface of functional layer  30  constitute boundary face  44 . Boundary faces  43  and  44  extend in first development direction X. First layer  31  is not in contact with third layer  33 . When the sample is developed in second development direction Y, second developing solvent  71  moves from functional layer  30  to first layer  31  through boundary face  44 . Second developing solvent  71  moves from first layer  31  to second layer  32  through boundary face  43 . Due to space  50 , second developing solvent  71  does not directly move to second layer  32  from functional layer  30 . Therefore, the multiple components located in first layer  31  can easily move to second layer  32 . 
     TLC plate  710  can be manufactured in such a way that second layer  32  and functional layer  30  are formed on substrate  10 , and then, first layer  31  is formed on second layer  32  and functional layer  30 . As a method for forming first layer  31  on second layer  32  and functional layer  30 , the methods described above as examples of the method for forming first layer  31  on substrate  10  in the first exemplary embodiment can be used, for example. In TLC plate  710 , first layer  31  is formed after the formation of second layer  32 , third layer  33 , and functional layer  30 , whereby separation layer  21  can be easily manufactured. 
     Another Modification of Second Exemplary Embodiment 
     As shown in  FIG. 10D , second layer  32  may be in contact with functional layer  30 . In TLC plate  720 , a lateral surface of second layer  32  and a lateral surface of functional layer  30  constitute boundary face  45 . First layer  31  is disposed on second layer  32  and functional layer  30 . A lower surface of first layer  31  and upper surfaces of second layer  32  and functional layer  30  constitute boundary face  46 . First layer  31  is not in contact with third layer  33 . Boundary faces  45  and  46  extend in first development direction X. When the sample is developed in second development direction Y, the developing solvent moves from functional layer  30  to second layer  32  through boundary face  45 . 
     TLC plate  720  is manufactured in the same manner as TLC plate  710 . In TLC plate  720 , first layer  31  is formed after the formation of second layer  32 , third layer  33 , and functional layer  30 , whereby separation layer  21  can be easily manufactured. 
     In TLC plate  720 , first developing solvent  70  is difficult to penetrate into each of second layer  32  and functional layer  30 . Therefore, when being developed by first developing solvent  70 , the multiple components included in sample  60  are held in first layer  31 . Then, TLC plate  720  is brought into contact with second developing solvent  71 . At that time, second layer  32  and functional layer  30  are both in contact with second developing solvent  71 . Therefore, the porous bodies constituting second layer  32  and functional layer  30  get wet with second developing solvent  71 . In this case, the multiple components held in first layer  31  tend to move to second layer  32  or functional layer  30  through boundary face  46 . That is, when the porous bodies constituting second layer  32  and functional layer  30  get wet, the multiple components tend to move in the thickness direction of separation layer  21 . This tendency is significant when alcohol is used as the organic solvent contained in second developing solvent  71 . Due to the movement of the multiple components to second layer  32  or functional layer  30 , the multiple components can be developed in second development direction Y. 
     Eighth Exemplary Embodiment 
     As shown in  FIGS. 11A and 11B , in TLC plate  800  according to the present third exemplary embodiment, first layer  31  is disposed on second layer  32 . Second layer  32  and third layer  33  are disposed on substrate  10 . In other words, only second layer  32  and third layer  33  are in contact with substrate  10 . Second layer  32  extends from an end face of substrate  10  in second development direction Y to boundary face  41  between second layer  32  and third layer  33 . A lower surface of first layer  31  and an upper surface of second layer  32  are in contact with each other. First layer  31  and second layer  32  constitute boundary face  47 . Boundary face  47  extends in first development direction X. First layer  31  is located between one end  32   a  and other end  32   b  of second layer  32  in second development direction Y. First layer  31  is not in contact with third layer  33 . 
     A distance from one end  32   a  of second layer  32  to first layer  31  in second development direction Y is equal to a value that can be assumed by length L 17  of functional layer  30  in TLC plate  700 . A distance from first layer  31  to other end  32   b  of second layer  32  in second development direction Y is equal to a value that can be assumed by length L 13  of second layer  32  in TLC plate  600 . 
     TLC plate  800  can be manufactured in such a way that second layer  32  is formed on substrate  10 , and then, first layer  31  is formed on second layer  32 . As a method for forming second layer  32  on substrate  10  and a method for forming first layer  31  on second layer  32 , the methods exemplified in the sixth exemplary embodiment can be used, for example. In TLC plate  800 , first layer  31  is formed after the formation of second layer  32 , whereby separation layer  22  can be easily manufactured. 
     In TLC plate  800 , first developing solvent  70  is difficult to penetrate into second layer  32 . Therefore, when being developed by first developing solvent  70 , the multiple components included in sample  60  are held in first layer  31 . Then, TLC plate  800  is brought into contact with second developing solvent  71 . At that time, second layer  32  is in contact with second developing solvent  71 . Thus, the porous body constituting second layer  32  gets wet with second developing solvent  71 . In this case, the multiple components held in first layer  31  tend to move to second layer  32  through boundary face  47 . That is, when the porous body constituting second layer  32  gets wet, the multiple components tend to move in the thickness direction of separation layer  22 . This tendency is significant when alcohol is used as the organic solvent contained in second developing solvent  71 . Due to the movement of the multiple components to second layer  32 , the multiple components can be developed in second development direction Y. 
     According to TLC plate  800 , the multiple components can move straight in second development direction Y in second layer  32  and third layer  33 . Specifically, when the multiple components are developed in second development direction Y, second developing solvent  71  is brought into contact with one end  32   a  of second layer  32 . In this case, a gradient may occur in a movement distance of second developing solvent  71  in second development direction Y. When the multiple components are developed with the gradient occurring in the movement distance, the multiple components may move in second layer  32  and third layer  33  in a direction different from second development direction Y. However, when the gradient occurs in the movement distance, a portion of second developing solvent  71  moves in first development direction X as well as in second development direction Y. Therefore, the gradient in the movement distance of second developing solvent  71  is reduced, as second developing solvent  71  moves in second layer  32 . In TLC plate  800 , when second developing solvent  71  moves from one end  32   a  of second layer  32  to first layer  31 , the gradient in the movement distance of second developing solvent  71  in second development direction Y is reduced. Thus, the multiple components can move straight in second development direction Y in second layer  32  and third layer  33 . 
     Ninth Exemplary Embodiment 
     The TLC plate may further include a pair of electrodes. In  FIGS. 12A  and  12 B, TLC plate  900  has a pair of electrodes  55 . The pair of electrodes  55  is disposed at both ends of first layer  31  in first development direction X. The pair of electrodes  55  is disposed on first layer  31 . If a voltage is applied to the pair of electrodes  55  with first layer  31  being impregnated with first developing solvent  70 , current flows through first layer  31 . A structure of TLC plate  900  is the same as the structure of TLC plate  800  in the eighth exemplary embodiment except for the pair of electrodes  55 . Note that an average pore diameter of the porous material constituting first layer  31  of TLC plate  900  may be within a range from 0.1 μm to 100 μm. With this configuration, the multiple components included in the sample can be easily electrophoresed in first layer  31 . 
     The pair of electrodes  55  is not particularly limited, as long as they can apply a voltage. The pair of electrodes  55  may be formed from at least one metal selected from the group consisting of platinum, gold, copper, and aluminum, for example. 
     Next, a sample analysis method using TLC plate  900  will be described. 
     First, sample  60  is placed on first layer  31  of separation layer  22  of 
     TLC plate  900 , as shown in  FIG. 13A . When sample  60  is placed on first layer  31 , sample  60  penetrates into first layer  31 , so that circular spot  61  is formed. Sample  60  is an aqueous solution containing a plurality of proteins, for example. In first layer  31 , a position where sample  60  is to be placed is not particularly limited. Sample  60  may be placed on a middle point of first layer  31  in first development direction X. In this case, the multiple components included in sample  60  can be quickly separated from each other by electrophoresis of the multiple components. First layer  31  is impregnated with first developing solvent  70  in advance. First developing solvent  70  is typically the same as that used in the first exemplary embodiment. 
     Then, a voltage is applied to electrodes  55  on TLC plate  900  as shown in  FIG. 13B . The multiple components included in sample  60  are electrophoresed in first development direction X. The voltage can be applied by power source  80 . Power source  80  is an AC-to-DC converter, a power generating device, or a battery, for example. The electrophoresis may be conducted inside an analyzing device. The multiple components are separated from each other in first layer  31  based on isoelectric point or molecular weight of each component. Due to the electrophoresis of the multiple components in first development direction X, spots  66 ,  67 ,  68 , and  69  are newly generated. 
     Then, as shown in  FIG. 13C , TLC plate  900  is placed in container  76  with end  32   a  of second layer  32  in second development direction Y being directed downward. Container  76  contains second developing solvent  71 . Container  76  and second developing solvent  71  are typically the same as those used in the sixth exemplary embodiment. 
     When TLC plate  900  is placed in container  76 , end  32   a  of second layer  32  is in contact with second developing solvent  71 . At that time, first layer  31  is impregnated with first developing solvent  70 . The liquid level of second developing solvent  71  is set to prevent direct contact between second developing solvent  71  and spots  66 ,  67 ,  68 , and  69 . The porous body constituting second layer  32  gets wet with second developing solvent  71  by second developing solvent  71 . Thus, the multiple components move from first layer  31  to second layer  32 . Due to capillary force, second developing solvent  71  moves in second development direction Y from end  32   a  of second layer  32 . When second developing solvent  71  is brought into contact with the multiple components located in spots  66 ,  67 ,  68 , and  69 , the multiple components are dissolved into second developing solvent  71 . The multiple components dissolved in second developing solvent  71  move in second development direction Y along with second developing solvent  71 . The multiple components which are not separated from each other in first layer  31  are separated from each other in second layer  32 . The multiple components which are not separated from each other in second layer  32  are separated from each other in third layer  33 . 
     Second layer  32  of TLC plate  900  includes a hydrophobic porous body. First developing solvent  70  hardly penetrates into second layer  32 . That is, pores in the porous body constituting second layer  32  hardly contain first developing solvent  70  after the electrophoresis of the multiple components included in sample  60 . Third layer  33  is not in contact with first layer  31 . Therefore, first developing solvent  70  hardly penetrates into third layer  33 . That is, pores in the porous body constituting third layer  33  hardly contain first developing solvent  70  after the electrophoresis of the multiple components included in sample  60 . Therefore, it is unnecessary to dry second layer  32  and third layer  33  after the multiple components included in the sample are developed in first development direction X. TLC plate  900  can be brought into contact with second developing solvent  71  with first layer  31  being impregnated with first developing solvent  70 . In other words, it is unnecessary to heat TLC plate  900  to a temperature higher than room temperature or leave TLC plate  900  in an atmosphere with a pressure lower than atmospheric pressure, during a period from when the voltage applied to electrodes  55  is removed till TLC plate  900  is brought into contact with second developing solvent  71 . According to TLC plate  900 , the multiple components can be developed in second development direction Y just after the development of the multiple components in first development direction X. Thus, the multiple components can be separated from each other more easily and more quickly. Note that, with the sample analysis method in the present exemplary embodiment, separation layer  22  may be dried before TLC plate  900  is brought into contact with second developing solvent  71 . 
     With the sample analysis method according to the present exemplary embodiment, it is unnecessary to change the orientation of TLC plate  900 . Therefore, the analyzing device used for the sample analysis method according to the present exemplary embodiment does not need a mechanism for changing the orientation of the TLC plate. 
     As a method for detecting the positions of the multiple components and a method for identifying each of the multiple components in the sample analysis method according to the present exemplary embodiment, the methods described in the first exemplary embodiment can be used. 
     Tenth Exemplary Embodiment 
     As shown in  FIGS. 14A and 14B , TLC plate  1000  according to the present tenth exemplary embodiment is obtained by further providing fourth layer  34  to nth layer  35  to the configuration of TLC plate  700  in the second exemplary embodiment. Each of fourth layer  34  to nth layer  35  induces an interaction different from the interactions induced by second layer  32  and third layer  33 , with respect to multiple components included in a sample. Therefore, the multiple components which are not separated from each other in second layer  32  and third layer  33  are separated from each other in fourth layer  34  to nth layer  35 . 
     Each of fourth layer  34  to nth layer  35  has a rectangular shape in a plan view. Each of fourth layer  34  to nth layer  35  extends in second development direction Y. The n is an integer equal to or greater than 4. The n is an integer from 5 to 10, for example. Each of fourth layer  34  to nth layer  35  is in contact with the corresponding one of third layer  33  to (n-1)th layer (not shown). When separation layer  23  is viewed in a plan view, one side of each of fourth layer  34  to nth layer  35  is in contact with one side of the corresponding one of third layer  33  to (n-1)th layer. The length of one side of each of fourth layer  34  to nth layer  35  is equal to the length of one side of the corresponding one of third layer  33  to (n-1)th layer. Functional layer  30  and first layer  31  to nth layer  35  are arrayed in this order in second development direction Y. 
     In the present exemplary embodiment, functional layer  30  and first layer  31  to nth layer  35  are disposed on substrate  10 . In other words, functional layer  30  and first layer  31  to nth layer  35  are in contact with substrate  10 . A lateral surface of each of first layer  31  to nth layer  35  is in contact with a lateral surface of the corresponding one of functional layer  30  and first layer  31  to (n-1)th layer. When the multiple components are developed in second development direction Y, a developing solvent sequentially moves in the order of functional layer  30  and first layer  31  to nth layer  35 . 
     Each of fourth layer  34  to nth layer  35  includes a porous body. The porous body constituting each of fourth layer  34  to nth layer  35  may be the same as any of those described as examples of the porous body constituting first layer  31  or second layer  32 . When the multiple components included in the sample are developed in first development direction X by electrophoresis, each of fourth layer  34  to nth layer  35  may include a hydrophilic porous body. An average pore diameter of each of fourth layer  34  to nth layer  35  may be within a range from 0.01 μm to 100 μm. When the porous body is an aggregate of inorganic particles, an average particle diameter of the inorganic particles may be within a range from 1 μm to 100 μm. Fourth layer  34  to nth layer  35  may further include any of the additives mentioned above. 
     TLC plate  1000  satisfies at least one requirement selected from among a requirement in which second layer  32  to nth layer  35  are different in composition and a requirement in which second layer  32  to nth layer  35  are different in structure. Accordingly, second layer  32  to nth layer  35  induce different interactions with the multiple components included in the sample. “Second layer  32  to nth layer  35  being different in structure” means that second layer  32  to nth layer  35  are different in at least one factor selected from among an average pore diameter of the porous body, a void ratio of the porous body, and an average particle diameter of the material of the porous body, for example. The multiple components are separated from each other in each of second layer  32  to nth layer  35 . 
     TLC plate  1000  may not satisfy the above-mentioned requirement depending on the multiple components included in sample  60 . For example, at least two layers selected from second layer  32  to nth layer  35  may have the same composition and the same structure, as long as they are not in contact with each other. 
     Hydrophobic property may be improved in the order of second layer  32  to nth layer  35 . Hydrophobic property may be decreased in the order of second layer  32  to nth layer  35 . At least one factor selected from among the average pore diameter, the void ratio, and the average particle diameter of the material of the porous body constituting each of the second layer  32  to nth layer  35  may be increased in the order of second layer  32  to nth layer  35 . At least one factor selected from among the average pore diameter, the void ratio, and the average particle diameter of the material of the porous body constituting each of the second layer  32  to nth layer  35  may be decreased in the order of second layer  32  to nth layer  35 . The content of additives may be increased in the order of second layer  32  to nth layer  35 . The content of additives may be decreased in the order of second layer  32  to nth layer  35 . 
     The length of each of fourth layer  34  to nth layer  35  in second development direction Y is equal to a value that can be assumed by length L 14  of third layer  33  of TLC plate  600 . The lengths of second layer  32  to nth layer  35  in second development direction Y may be the same as or different from each other. 
     As a method for forming fourth layer  34  to nth layer  35  on substrate  10 , the methods described above as examples of the method for forming first layer  31  on substrate  10  and the method for forming second layer  32  on substrate  10  in the first exemplary embodiment can be used, for example. 
     Modification of Tenth Exemplary Embodiment 
     As shown in  FIG. 14C , first layer  31  may be disposed on second layer  32  and functional layer  30 . A structure of TLC plate  1010  is the same as the structure of TLC plate  710  in the seventh exemplary embodiment except for fourth layer  34  to nth layer  35 . 
     Another Modification of Tenth Exemplary Embodiment 
     As shown in  FIG. 14D , second layer  32  may be in contact with functional layer  30 . A structure of TLC plate  1020  is the same as the structure of TLC plate  720  in the seventh exemplary embodiment except for fourth layer  34  to nth layer  35 . 
     INDUSTRIAL APPLICABILITY 
     The technique disclosed in the present specification is useful for protein analysis or the like. 
     REFERENCE MARKS IN THE DRAWINGS 
     
         
           10 : substrate 
           20 ,  21 ,  22 ,  23 : separation layer 
           30 : functional layer 
           31 : first layer 
           32 : second layer 
           33 : third layer 
           55 : electrode 
           60 : sample 
           91 ,  92 ,  93 : end face 
           100 ,  200 ,  210 ,  220 ,  300 ,  400 ,  500 ,  600 ,  700 ,  710 ,  720 ,  800 ,  900 ,  1000 , 
           1010 ,  1020 : TLC plate (thin layer chromatography plate) 
         X: first development direction 
         y: second development direction