WO2011149041A1 - 薄層クロマトグラフィーによる試料の検出方法、薄層クロマトグラフィープレート、及びその製造方法 - Google Patents
薄層クロマトグラフィーによる試料の検出方法、薄層クロマトグラフィープレート、及びその製造方法 Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/02—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material
- B01J20/04—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising compounds of alkali metals, alkaline earth metals or magnesium
- B01J20/045—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising compounds of alkali metals, alkaline earth metals or magnesium containing sulfur, e.g. sulfates, thiosulfates, gypsum
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/3042—Use of binding agents; addition of materials ameliorating the mechanical properties of the produced sorbent
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/92—Construction of the plate
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/92—Construction of the plate
- G01N30/93—Application of the sorbent layer
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/94—Development
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/20—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/265—Adsorption chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/94—Development
- G01N2030/945—Application of reagents to undeveloped plate
Definitions
- the present invention relates to a thin-layer chromatography plate (hereinafter also referred to as “TLC plate”) having two kinds of separating agent layers having different optical responsiveness to ultraviolet light or a coloring reagent, a production method thereof, and a usage method thereof.
- TLC plate thin-layer chromatography plate
- TLC thin layer chromatography
- separation agents for optical isomers separation agents containing polysaccharide derivatives such as polysaccharide phenyl esters are known.
- the detection target component may not be detected by ultraviolet irradiation or color developing treatment of the coloring reagent.
- a TLC plate for example, a TLC plate in which a filler layer for optical resolution is provided on a partial surface of a support on a flat plate is known (for example, see Patent Document 1).
- an optical resolution filler layer and a silica gel-based filler layer are formed on the surface of a single support, and components (spots) separated by the optical resolution filler layer are further separated into a silica gel-based filler layer.
- the component to be detected is detected by irradiating with ultraviolet rays in the silica gel-based filler layer or by performing a color developing treatment of the color developing reagent.
- the extract component that is easily adsorbed by the optical resolution filler layer is sufficiently transferred from the optical resolution filler layer to the silica gel type filler layer compared to the raffinate component that is difficult to be adsorbed by the filler layer. Otherwise, the silica gel-based filler layer may not be sufficiently detected, and the separation state in the optical resolution filler layer may not be confirmed. For this reason, the TLC plate may not be able to obtain necessary information for the purpose of studying the optical resolution conditions by other chromatography such as column chromatography. ing.
- the present invention provides a TLC plate and a TLC that can detect separation of components in a sample by a separating agent layer that cannot detect separation of components in the sample by optical response.
- the present inventors diligently studied the use of a separating agent having optical response to ultraviolet light or a color developing reagent in TLC, and a spot developed by the first separating agent layer having the specific optical response was identified as a spot.
- a spot developed by the first separating agent layer having the specific optical response was identified as a spot.
- the present invention uses a TLC plate having a first separating agent layer and a second separating agent layer adjacent thereto, and detects a component in a sample having optical responsiveness to ultraviolet light or a coloring reagent. And in the first separating agent layer, developing the sample in a direction along the boundary between the first and second separating agent layers, drying the mobile phase of at least the second separating agent layer, The step of changing the direction of the TLC plate to develop the spot in the first separating agent layer on the second separating agent layer, and the spot moved to the second separating agent layer by ultraviolet irradiation or color developing treatment of the coloring reagent Detecting, wherein the first separating agent layer is formed of the first separating agent having optical response, and the second separating agent layer is different from the first separating agent in the optical response. Formed with a second separating agent The optical response of the component is to provide a method which is the same as the optical response of the first separating agent.
- the present invention also includes a first separating agent layer having a first direction as a developing direction, and a second separating agent layer adjacent to the first separating agent layer in a second direction orthogonal to the first direction.
- the first separating agent layer is formed of a first separating agent having optical responsiveness to ultraviolet light or a coloring reagent, and the second separating agent layer is different from the first separating agent.
- the spot is developed in the first direction in the first separating agent layer
- the spot movement in the first direction in the first separating agent layer is formed by the second separating agent having optical response.
- a TLC plate in which the ratio of the distance W2 of the spot in the second direction in the first separating agent layer to the distance W1 is ⁇ 0.5 or less.
- the present invention also provides the above method and TLC plate, wherein the first separating agent is a separating agent for optical isomers.
- the separating agent for optical isomers may be any of an aromatic ester group, an aromatic carbamoyl group, an aromatic ether group, and a carbonyl group in which a part of or all of the polysaccharide and the hydroxyl group or amino group of the polysaccharide are replaced.
- TLC plate comprising a polysaccharide derivative comprising heel.
- the present invention also relates to a method for producing a TLC plate having a first separating agent layer and a second separating agent layer adjacent thereto, the first separating agent having optical responsiveness to ultraviolet light or a coloring reagent. And a step of applying a first slurry containing a coating solvent to a support to form a first separating agent layer, and a second separating agent having optical response different from the first separating agent And applying a second slurry containing a coating solvent to a support to form a second separating agent layer adjacent to the first separating agent layer, and in the first and second slurries A method of using the same type of coating solvent is provided.
- the spot developed in the first direction in the first separating agent layer is transferred to the second separating agent layer while maintaining the positional relationship of the spots in the first direction, an optical response is obtained. Then, it is possible to detect separation of components in the sample by a separating agent layer that cannot detect separation of components in the sample.
- the spot movement width allowed in the development direction in the first separation agent layer of the TLC plate is specified within a predetermined range, the development direction in the first separation agent layer is increased. The separation state of these components can be accurately grasped in the second separating agent layer.
- the present invention provides a TLC plate having good spot mobility from the first separating agent layer to the second separating agent layer because the types of the first and second slurry coating solvents are uniform. be able to.
- FIG. 1 It is a figure which shows the state which spotted the sample solution to an example of the TLC plate of this invention. It is a figure which shows the state which expand
- FIG. 1 In the TLC plate of FIG. 1, it is a figure which shows the state which develops the spot developed on the 1st separating agent layer 1 toward the 2nd separating agent layer 2.
- the method for detecting a sample in the present invention uses a TLC plate having a first separating agent layer and a second separating agent layer adjacent to the first separating agent layer, and the components in the sample having optical responsiveness to ultraviolet light or a coloring reagent. It is a method of detection.
- the detection method of the present invention in the first separating agent layer, the step of developing the sample in a direction along the boundary between the first and second separating agent layers and the mobile phase of at least the second separating agent layer are dried.
- a step of changing the direction of the TLC plate to develop the spot in the first separating agent layer on the second separating agent layer, and the spot moved to the second separating agent layer is irradiated with ultraviolet rays or a coloring reagent. Detecting by color development.
- the step of developing the sample in the direction along the boundary between the first and second separating agent layers is performed by the first separating agent along the second separating agent layer. It can be performed in the same manner as normal TLC except that the sample is developed only in the layer. That is, in the first step, the sample is spotted on the first separating agent layer, the sample spot is dried as necessary, and the direction along the boundary between the first and second separating agent layers (" The first direction is also referred to as the “first direction”, and the ends of the first and second separating agent layers are immersed in the mobile phase with the end of the separating agent layer on which the sample is deposited facing down. This can be done by developing the sample in the agent layer.
- the step (second step) of drying the mobile phase of at least the second separating agent layer can be performed in the same manner as the mobile phase drying in normal TLC.
- the second step if the second separating agent layer is dried, it is possible to move the spot to the second separating agent layer, but not only the second separating agent layer but also the second separating agent layer. It is preferable to dry the mobile phase of one separating agent layer from the viewpoint of preventing the spread of spots in the first separating agent layer and preserving the positional relationship.
- the second step is preferably air-dried in the vicinity of the local exhaust device from the viewpoint of preventing fire and poisoning.
- the second step may be performed under reduced pressure.
- the step (third step) of changing the direction of the TLC plate to develop the spot in the first separating agent layer on the second separating agent layer (third step), the spot in the first separating agent layer to the second separating agent layer
- the direction of the TLC plate is changed so that the mobile phase develops in a direction crossing the boundary between the first and second separating agent layers (also referred to as “second direction”), This can be done by immersing the end of the plate in the mobile phase and developing the mobile phase so that the spot in the first separating agent layer moves over the boundary to the second separating agent layer.
- the second direction can be determined from a range of greater than 0 ° and less than or equal to 90 ° with respect to the first direction. From the viewpoint of the accuracy of storing the positional relationship of spots in the first mobile phase, the second direction 90 ° is preferred, 60 to 90 ° is more preferred, and 90 ° is even more preferred.
- the mobile phase in the first step and the mobile phase in the third step may be the same or different.
- the spot in the third step, if a mobile phase having a higher polarity than the mobile phase in the first step is used, the spot can be moved more quickly from the first separating agent layer to the second separating agent layer. it can.
- the mobile phase water, an aqueous salt solution, an organic solvent, or a mixed solvent thereof can be used.
- One or more organic solvents may be used.
- the aqueous salt solution include an aqueous copper sulfate solution and a perchlorate.
- the organic solvent include alcohols such as methanol, ethanol and isopropanol, ketones such as acetone and ethyl methyl ketone, ethers such as tetrahydrofuran and dioxane, nitriles such as acetonitrile, sulfoxides such as dimethyl sulfoxide, sulfones such as sulfolane, and the like.
- Esters such as ethyl acetate, amides such as dimethylformamide, hydrocarbons such as pentane, hexane, petroleum ether, aromatic hydrocarbons such as benzene, toluene, xylene, and methylene chloride, chloroform, bromoform, And halogen-containing compounds such as chlorobenzene and bromobenzene.
- an appropriate amount of acid or alkali may be mixed in the mobile phase from the viewpoint of stabilizing the component to be detected.
- the acid include acetic acid, propionic acid, and tetrafluoroacetic acid
- examples of the alkali include diethylamine, monoethanolamine, and triethylamine.
- the step (fourth step) of detecting the spot that has moved to the second separating agent layer by ultraviolet irradiation or color developing treatment of the coloring reagent is performed in TLC, except that the spot in the second separating agent layer is to be detected. It can be carried out in the same manner as in the usual method for detecting spots by irradiating with ultraviolet rays or coloring treatment with a coloring reagent.
- the first separating agent layer is formed of a first separating agent having optical responsiveness to ultraviolet light or a coloring reagent.
- the optical responsiveness to ultraviolet rays refers to light emission or absorption of ultraviolet rays such as fluorescence.
- the optical responsiveness to the coloring reagent refers to color development by the coloring reagent.
- the first separating agent layer can be formed in the same manner as a thin layer of a normal TLC plate except that it is formed of the first separating agent.
- the first separating agent includes a separating agent component having the optical response.
- a first separating agent examples include a carrier-type separating agent composed of a carrier and the separating agent component supported or bonded to the carrier, and particles composed of the separating agent component.
- the first separating agent is preferably a separating agent for optical isomers, in which simple detection of a sample by TLC is generally difficult due to the performance and physical properties of the separating agent.
- the carrier is preferably a porous body from the viewpoint of improving the separation performance.
- the carrier include synthetic polymers such as cross-linked polystyrene, cross-linked acrylic polymer, and epoxy polymer, cellulose and cross-linked cellulose reinforced by cross-linking, cross-linked agarose, cross-linked dextran, and cross-linked mannan cross-linked polysaccharides, and , Alumina, silica gel, mesoporous silica gel, zeolite, diatomaceous earth, fused silica, viscosity mineral, zirconia, metals and other inorganic materials.
- separating agent component in the first separating agent either a low molecular separating agent or a high molecular separating agent having optical response can be used.
- the low molecular separation agent include a ligand exchange type separation agent, a charge transfer ( ⁇ - ⁇ ) type separation agent, a hydrogen bond type separation agent, an inclusion type separation agent, and an ion bond type separation agent.
- examples thereof include a separating agent, an intercalating type separating agent, a crown ether or a derivative thereof, and a cyclodextrin or a derivative thereof.
- the polymer separating agent include polysaccharide derivatives, polyamides, polymethacrylic acid esters, polyacrylamides, proteins, and tartaric acid derivatives.
- polysaccharide derivatives examples include aromatic ester groups, aromatic carbamoyl groups, aromatic ether groups, and carbonyls, which are used in separating agents for optical isomers, for example, by replacing part or all of hydroxyl groups or amino groups of polysaccharides and polysaccharides.
- examples thereof include polysaccharide derivatives composed of any of the groups, and examples thereof include phenyl carbamate derivatives of cellulose, phenyl ester derivatives of cellulose, phenyl carbamate derivatives of amylose, and phenyl ester derivatives of amylose.
- the phenyl group in these derivatives may have one or more substituents selected from the group consisting of hydrocarbons having 1 to 20 carbon atoms and halogens.
- the second separating agent layer is formed of a second separating agent having an optical response different from that of the first separating agent.
- the different optical responsiveness means that one optical response and the other optical response due to ultraviolet irradiation or color developing treatment of the coloring reagent are different from each other to the extent that they can be optically discriminated by color and brightness.
- the second separating agent may or may not have separation ability for the components in the sample. From the viewpoint of accurate detection of the positional relationship between spots in the first separating agent layer, it is preferable that the second separating agent does not have separability with respect to components in the sample. Examples of such a second separating agent include the carriers and separating agents described above, but a carrier is preferable.
- the particle size of the first and second separation agents can be determined according to the purpose of the detection method of the present invention, and can be applied to medium pressure chromatography, for example, for studying the conditions of medium pressure column chromatography. From the viewpoint of obtaining satisfactory results, the thickness is preferably 10 ⁇ m or more, more preferably 10 to 100 ⁇ m, and even more preferably 20 to 100 ⁇ m.
- the particle size of each separating agent may be the same or different. As the particle size of each separating agent, an average particle size measured by a normal particle size measuring device can be adopted, but it may be a catalog value.
- the second separating agent preferably further contains a fluorescent indicator from the viewpoint of clearer detection.
- a fluorescent indicator a known fluorescent indicator can be used, and examples thereof include magnesium tungstate and manganese-containing zinc silicate.
- the content of the fluorescent indicator can be determined within a range where the components can be separated, and is generally 0.1 to 5 parts by mass with respect to 100 parts by mass of the second separating agent.
- the coloring reagent when the component is detected by optical responsiveness due to the coloring reagent, a known technique can be adopted for the coloring reagent and its coloring process.
- a coloring reagent include anisaldehyde solution, phosphomolybdic acid solution, iodine, ninhydrin solution, chameleon solution, DNPH solution, manganese chloride solution, and bromocresol green solution.
- the coloring process include a process in which a coloring reagent is applied to the TLC plate by application, dispersion, or exposure, and the TLC plate is heated as necessary to develop a color.
- the optical response of the component in the sample is the same as the optical response of the first separating agent.
- the same optical response means that one optical response and the other optical response cannot be optically distinguished by color or brightness.
- the components in the sample that are separated by at least the first separation agent but cannot be optically detected on the first separation agent are first analyzed by TLC. Separation by the separating agent can be easily detected.
- the TLC plate of the present invention described later can be suitably used as the TLC plate in the detection method of the present invention.
- the TLC plate of the present invention has a flat support, a first separating agent layer formed on the surface of the supporting member, and a second separating agent layer adjacent thereto.
- a flat plate generally used for a TLC plate can be used.
- a support examples include a flat plate made of glass, resin, metal, or paper.
- the shape of the support is not particularly limited, but from the viewpoint of simply performing the development of the sample on the first separating agent layer and the subsequent development of the spots from the first to the second separating agent layer in a preferred direction. It is preferably rectangular or square. And it is not particularly restricted but includes the size of the flat plate, the handling ease and deployment time is preferably from an economic point of view from 1 ⁇ 6,000 2, more preferably 2 ⁇ 410cm 2, 5 ⁇ More preferably, it is 75 cm 2 .
- the thickness of the flat plate is not particularly limited as long as it can be applied to the detection method of the present invention.
- the first and second separating agents two or more kinds of separating agents having different optical responsiveness can be used from among particulate separating agents having a particle size of 10 ⁇ m or more.
- the first separating agent layer is formed by applying a first slurry containing the first separating agent and a coating solvent to a support, and further drying the formed layer as necessary. Can be formed.
- the second separating agent layer is formed by applying a second slurry containing the second separating agent and a coating solvent to the support, and the formed layer is further dried as necessary. Can be formed.
- the first and second separating agent layers can be formed using a known method for producing a TLC plate. That is, the first and second separating agent layers are applied by applying the first and second slurries to the surface of the support using a spreader, or the first and second slurries are supported on the support. It can be formed by spraying on the surface.
- the order in which the first and second separating agent layers are formed is not particularly limited. After one layer is formed, the other layer may be formed, or both layers may be formed simultaneously.
- the coating solvent water, an organic solvent, and a mixed solvent thereof can be used.
- the organic solvent include alcohols such as methanol, ethanol and isopropanol, ketones such as acetone and ethyl methyl ketone, ethers such as tetrahydrofuran and dioxane, nitriles such as acetonitrile, sulfoxides such as dimethyl sulfoxide, sulfones such as sulfolane, and the like.
- Esters such as ethyl acetate, amides such as dimethylformamide, hydrocarbons such as pentane, hexane, petroleum ether, aromatic hydrocarbons such as benzene, toluene, xylene, methylene chloride, chloroform, bromoform, chlorobenzene, And halogen-containing compounds such as bromobenzene.
- the coating solvent is preferably a mixed solvent of a water-soluble organic solvent and water, more preferably a mixed solvent of alcohol and water, and further preferably a mixed solvent of ethanol and water.
- the alcohol content in the mixed solvent is preferably 0.1 to 50% by mass, more preferably 10 to 40% by mass, and further preferably 20 to 30% by mass.
- the types of the coating solvent are the same, and it is more preferable that the composition of the coating solvent is the same.
- the content of the coating solvent in the first and second slurries can be determined from the uniformity of the separating agent layer to be formed, the thickness of the layer, and an economic viewpoint. Any of these slurries is preferably 10 to 5,000 parts by weight, more preferably 50 to 1,000 parts by weight, and more preferably 100 to 300 parts by weight with respect to 100 parts by weight of the first or second separating agent. More preferably, it is part by mass.
- the first and second slurries preferably further contain a binder from the viewpoint of improving the strength of the separating agent layer to be formed.
- a binder a component that brings about a binding property to form a separating agent layer on the surface of the support can be used.
- binders include inorganic binders such as gypsum and colloidal silica, organic fibers such as microfibrillated cellulose, alkali water-soluble copolymers, thickeners such as hydroxyethyl cellulose and carboxymethyl cellulose, polyvinyl alcohol, acrylic An organic binder such as an acid may be used.
- the binder may be one type or two or more types.
- the content of the binder in the first and second slurries can be determined from the viewpoint of the rising speed of the mobile phase in the first or second separating agent layer in addition to the strength of the separating agent layer to be formed. From the viewpoint of improving the strength of the separating agent layer to be formed, it is preferable that the amount of the binder is large, but in general, the rate of increase of the mobile phase in the separating agent layer is slow in proportion to the increase in the amount of binder added. Become. In the first step, when the mobile phase rising rate in the second separating agent layer is significantly smaller than the mobile phase rising rate in the first separating agent layer, the sample spot is in the first direction.
- a phenomenon of rising while approaching the second separating agent layer may occur, and the separation of the sample by the first separating agent layer may not be analyzed correctly.
- the spot of the sample will be in the first direction relative to the first direction.
- a phenomenon of rising while moving away from the second separating agent layer occurs, and the separation of the sample by the first separating agent layer may not be correctly analyzed.
- the spot of the sample rises in the first direction (for example, vertical) as much as possible. It is preferable to do.
- the rising speed of the mobile phase in the first separating agent layer is higher than that in the second separating agent layer.
- the rising speed of the mobile phase is remarkably large, spot tailing occurs, and it may be difficult to detect the component in the second separating agent layer.
- the mobile phase in the first separating agent layer The rising speed is equal to the rising speed of the mobile phase in the second separating agent layer, or the rising speed of the mobile phase in the second separating agent layer relative to the rising speed of the mobile phase in the first separating agent layer. It is preferable to make the rising speed slightly smaller.
- the TLC plate in the present invention has a spot moving distance W1 in the first direction in the first separating agent layer when the spot is developed in the first direction in the first separating agent layer.
- the ratio (W2 / W1) of the movement distance W2 of the spot in the second direction in the first separating agent layer is ⁇ 0.5 or less.
- W1 is represented by the distance from the spotting position of the sample in the first separating agent layer to the tip of the spot.
- W2 is represented by the distance from the spotting position to the tip of the spot in the second direction.
- W2 / W1 is preferably ⁇ 0.3 or less from the viewpoint of correctly grasping the separation state in the first separating agent layer, more preferably a positive value from the viewpoint of suppressing tailing, and 0 More preferably, it is 1 or less.
- W2 is expressed by assuming that the spotting position of the sample is 0, the second separating agent layer side from the spotting position is positive, and the side opposite to the second separating agent layer from the spotting position is negative. .
- both an inorganic binder and an organic binder in both the first and second slurries from the viewpoint of improving the strength of the layer and controlling the rising speed of the mobile phase.
- Gypsum is preferably used as the inorganic binder
- carboxymethyl cellulose is preferably used as the organic binder.
- the content of the binder in the first and second slurries can be determined from the viewpoint of obtaining the strength of the layer to be formed and the appropriate range of W2 / W1 described above.
- the content of gypsum in the first slurry is preferably 0.1 to 50 parts by mass, more preferably 5 to 30 parts by mass with respect to 100 parts by mass of the first separating agent. More preferably, it is ⁇ 20 parts by mass.
- the content of the organic binder in the first slurry is preferably 0.1 to 50 parts by mass, and 0.5 to 10 parts by mass with respect to 100 parts by mass of the first separating agent. Is more preferably 1 to 3 parts by mass.
- the content of gypsum in the second slurry is preferably 0.1 to 50 parts by mass, preferably 1 to 30 parts by mass with respect to 100 parts by mass of the second separating agent. More preferably, it is ⁇ 8 parts by mass.
- the content of the organic binder in the second slurry is preferably 0.1 to 50 parts by mass, and 0.5 to 10 parts by mass with respect to 100 parts by mass of the second separating agent. Is more preferably 2 to 4 parts by mass.
- the strength of the first and second separating agent layers only needs to be strong enough to withstand the storage and use of TLC, for example, such that the separating agent layer does not peel or collapse during storage or use. .
- W2 / W1 can be confirmed, for example, by spotting and developing a colored sample in the first separating agent layer.
- W2 / W1 is not limited to the binder type and blending amount. It is also possible to adjust by the thickness of the first and second separating agent layers.
- the first and second slurries may further contain a dispersant from the viewpoint of uniformly dispersing the binder on the surface of the dispersant.
- a dispersant examples include styrene / maleic anhydride polymer.
- the thickness of the first and second separating agent layers can be appropriately determined according to the purpose of use of the TLC plate.
- the thickness of the separating agent layer is 0 if the obtained result is applied to medium pressure column chromatography, or the obtained result is used for reaction monitoring of asymmetric synthesis reaction or fractionation of a compound. 0.01 to 100 mm is preferable, 0.03 to 10 mm is more preferable, and 0.05 to 2 mm is even more preferable.
- the size of the TLC plate is not particularly limited, but can be appropriately determined according to the separating agent, the sample, and the mobile phase, and is preferably large from the viewpoint of separation of the components. From the viewpoint of simplicity, it is desirable that it is not too large. From such a viewpoint, the size of the first and second separating agent layers can be appropriately determined.
- the length (B) of the first separating agent layer in the first direction may be long enough to develop the sample in the first direction, and the first separating agent, sample, and It can be determined appropriately according to the mobile phase.
- the length B of the first separating agent layer in the first direction is preferably 1 to 30 cm, more preferably 3 to 20 cm, more preferably 5 to 12 cm from the viewpoints of operability and detection reliability. More preferably it is.
- the length (A1) in the second direction of the first separating agent layer is preferably 0.01 to 200 cm, and preferably 0.01 to 20 cm from the viewpoints of operability and detection reliability. Is more preferably 0.02 to 5 cm.
- the length (A2) of the second separating agent layer in the second direction is preferably 0.1 to 30 cm, and preferably 0.5 to 20 cm from the viewpoint of operability and detection reliability. More preferably, the thickness is 1 to 5 cm.
- the ratio of A2 to A1 is preferably large from the viewpoint of moving the spot from the first separating agent layer to the second separating agent layer by the development of the mobile phase. From the viewpoint of simplicity of detection, it is desirable that it is small. From these viewpoints, it is preferably 0.1 to 20, more preferably 0.1 to 10, and more preferably 0.1 to 5. Is more preferable.
- the ratio of B to A1 (B / A1) is preferably large from the viewpoint of separation of components in the sample in the first separating agent layer, and is desirably small to some extent from the viewpoint of operability. From these viewpoints, B / A1 is preferably 0.01 to 9,000, more preferably 0.2 to 4,000, and even more preferably 1 to 600.
- the ratio of B to the sum of A1 and A2 (B / (A1 + A2)) is desirably large from the viewpoint of separation of components in the sample in the first separating agent layer, and from the first separating agent layer to the second separating agent layer. From the viewpoint of movement of the spot to the separating agent layer, particularly the movement of the extract component and the operability, it is desirable that it is small. From these viewpoints, B / (A1 + A2) is preferably from 0.01 to 30, more preferably from 0.1 to 30, and even more preferably from 0.8 to 12.
- the shapes of the first and second separating agent layers are not particularly limited as long as they are adjacent to each other and satisfy the above-described length condition.
- the shapes of the first and second separating agent layers may be rectangular from the viewpoints of operability, clarification of the display of the positional relationship of spots in the first separating agent layer, and ease of formation of each layer.
- the shape of the first separating agent layer is a rectangle having the first direction as a longitudinal direction, and the second separating agent layer has the same length as the first separating agent layer in the first direction. It is more preferable that the rectangle has a size satisfying the ratio A2 / A1.
- the TLC plate of the present invention has a first separating agent layer 1 and a second separating agent layer 2 which are provided adjacent to the surface of a support (not shown) having a square shape.
- a glass plate is used as the support
- the first separating agent constituting the first separating agent layer 1 is a separating agent having the ability to separate components in the sample, for example, 3,5-dimethylphenyl of amylose.
- a separating agent for optical isomers formed by carrying or chemically bonding a carbamate derivative on silica gel is used, and the second separating agent constituting the second separating agent layer 2 is an optical material different from the first separating agent.
- a responsive separating agent such as silica gel, cellulose, or octadecyl silylated silica gel whose surface is modified with an octadecyl group is used.
- the first separating agent layer 1 is formed by applying a first slurry comprising a first separating agent, an aqueous ethanol solution, gypsum and carboxymethyl cellulose to the surface of the support.
- the second separating agent layer 2 comprises a second slurry comprising a second separating agent, an aqueous ethanol solution, gypsum and carboxymethyl cellulose, and a fluorescent indicator, and the first separating agent layer 1 on the surface of the support. It is formed by applying at a position adjacent to the film.
- the blending ratio of the binder in each of the first and second slurries is the same as when the sudan or 1,4-aminoazobenzene was developed as a standard sample in the first separating agent layer 1.
- the ratio (W2 / W1) of the distance W2 from the spotting position in the second direction to the tip of the raffinate component spot to the distance W1 from the spotting position in the first direction to the tip of the raffinate component spot is It is adjusted to be ⁇ 0.5 or less.
- the standard sample has a similar or characteristic structure (for example, the same or similar skeleton structure, etc.) from the known substances to the sample to be analyzed by the TLC plate.
- a colored compound can be appropriately selected.
- the particle sizes of the first and second separating agents are each 20 ⁇ m, and the thicknesses of the first and second separating agents are 150 ⁇ m, respectively.
- the first direction in the TLC plate is a direction along the boundary between the first and second separating agent layers 1 and 2 (the direction indicated by the arrow X in FIG. 1), and the second direction is the X direction.
- the direction is orthogonal (the direction indicated by the arrow Y in FIG. 1).
- the first separating agent layer 1 is formed in a rectangle whose longitudinal direction is the X direction.
- the second separating agent layer 2 is formed in a rectangular shape that shares one long side of the first separating agent layer 1.
- the length B of the first and second separating agent layers 1 and 2 in the X direction is 10 cm
- the length A1 of the first separating agent layer 1 in the Y direction is 2 cm
- the second separation in the same direction The length A2 of the agent layer 2 is 2.5 cm.
- the X direction is the vertical direction of the TLC plate, and a sample solution is spotted on the lower part of the first separating agent layer 1.
- the sample solution is, for example, a solution of a racemic volatile solvent of an optical isomer having ultraviolet absorptivity.
- the spotting of the sample solution is 1.5 cm from the lower edge of the TLC plate and the side edge of the first separating agent layer 1 not adjacent to the second separating agent layer 2 in the TLC plate, respectively. It is performed using a capillary or a micropipette so that the spot diameter is 2 mm or less. After spotting, the spot 3 of the sample solution is dried.
- the TLC plate spotted with the sample solution can accommodate the TLC plate and is accommodated in a container (developing tank) capable of accommodating a mobile phase (developing liquid) at a depth of less than 1 cm.
- a mobile phase for example, a mixed solvent of n-hexane and ethanol or a mixed solvent of n-hexane and ethyl acetate is used.
- the mobile phase is sucked up by the first and second separating agent layers 1 and 2 in the X direction, and the raffinate component and the extract component in the sample solution become the first separating agent layer as the mobile phase is sucked up.
- the spot R of the raffinate component and the spot E of the extract component are formed as shown in FIG.
- W1 is the distance of movement of the spot in the first direction X in the first separating agent layer 1, and the distance between the tip of the spot and the spot position of the sample solution in the first direction. It is represented by Further, as shown in FIG. 2, W2 is the moving distance of the spot in the second direction Y in the first separating agent layer 1, and in the second direction between the tip of the spot and the spotting position. Expressed in distance.
- the TLC plate is accommodated in the sealed container with the first separating agent layer 1 as the bottom and the Y direction as the vertical direction of the TLC plate.
- the mobile phase at this time may be the same as or different from the mobile phase used for the development of the previous sample.
- the mobile phase is sucked up in the Y direction from the first separating agent layer 1 to the second separating agent layer 2, and as the mobile phase is sucked up, as shown in FIG.
- the raffinate component and the extract component move to the second separating agent layer 2 and cannot be visually confirmed, but the spot R of the raffinate component and the spot E of the extract component are formed on the second separating agent layer 2. .
- the TLC plate is taken out from the sealed container, and the mobile phase of the first and second separating agent layers 1 and 2 is cooled by cold air. Is dried and the spots R and E in the second separating agent layer 2 are fixed.
- the spots R and E and the first separating agent layer 1 have a similar optical response (for example, as shown in FIG. 5 (for example, Observed as a dark area due to absorption of ultraviolet light or a colored area due to coloration of the coloring reagent).
- the first step is performed without changing the direction of the TLC plate in the third step.
- the spot of the separating agent layer is developed on the second separating agent layer (that is, in the third step, the second direction is determined from a range from 0 ° to 90 ° with respect to the first direction.
- the component in the sample by the first separating agent layer cannot be detected by optical response even if the spot of one separating agent layer is developed on the second separating agent layer) Separation can be detected.
- the second step can be omitted, and the present invention includes such a case.
- Example 1 Preparation of TLC plate (adjustment of first separating agent layer) Daicel Chemical Industries, Ltd. CHIRALPAK IA (Registered Trademark) filler (6 g), gypsum 0.89 g, 2% CMC (carboxymethylcellulose) 1110 (Daicel Chemical Industries, Ltd.) ) 6 g of an aqueous solution was added to a mixed solution of 3.03 g of water and 3.01 g of ethanol, and stirred sufficiently while irradiating ultrasonic waves to obtain a slurry.
- CHIRALPAK IA Registered Trademark
- the TLC plate coated with the slurry is air-dried and vacuum-dried at 60 ° C. for 3 hours while being drawn with a vacuum pump.
- the IA filler layer is used as the first separating agent layer, and the silica gel layer is used as the second separating agent layer.
- a TLC plate was obtained.
- Example 2 Example of optical resolution of trans-stilbenzene oxide About 1 ⁇ L of a 1% ethyl acetate solution of trans-stilbenzene oxide (t-SO) was used with the longitudinal direction of the TLC plate prepared in Example 1 as vertical. It was spotted at a position about 1.5 cm from the bottom of the time. In a developing tank containing a mixed solvent containing n-hexane and ethanol in a volume ratio of 9: 1, the TLC plate is accommodated with the sample spot facing down, and the sample is developed in the longitudinal direction of the IA separating agent layer. It was.
- t-SO trans-stilbenzene oxide
- the TLC plate is once dried with cold air, rotated at 90 ° C. so that the IA separating agent layer is located below, and the TLC plate is accommodated in the developing layer, and the IA separating agent layer using ethanol.
- the sample was further developed from to the silica gel layer.
- the TLC plate was dried with cold air, irradiated with ultraviolet rays, and the positions of the raffinate component and extract component spots were confirmed in the silica gel layer.
- the distance from the spotting position to the reaching position of the developing liquid, and in the second separating agent layer, the distance from the spotting position of the sample in the first developing direction to the center of each spot From the above, the Rf value of each spot was determined.
- the Rf value of the raffinate component of t-SO was 0.67
- the Rf value of the extract component was 0.58.
- Example 3 Example of optical resolution of flavanone About 1 ⁇ L of 1% ethyl acetate solution of flavanone (FLV) was spotted on the TLC plate prepared in Example 1, and developed in the same manner as in Example 2.
- FLV flavanone
- the TLC plate was dried with cold air, irradiated with ultraviolet rays, and the positions of the raffinate component and extract component spots were confirmed in the silica gel layer.
- the Rf value was determined in the same manner as in Example 2, the Rf value of the raffinate component of FLV was 0.40, and the Rf value of the extract component was 0.29.
- Example 4 Example of optical resolution of benzoin ethyl ether About 1 ⁇ L of a 1% ethyl acetate solution of benzoin ethyl ether was spotted on the TLC plate prepared in Example 1, and n-hexane and ethyl acetate in a volume ratio of 9: 1 was used in the same manner as in Example 2 except that the mixed solvent contained in 1 was used as the developing solution in the first development and ethyl acetate was used in the developing solution in the second development.
- the TLC plate was dried with cold air, irradiated with ultraviolet rays, and the positions of the raffinate component and extract component spots were confirmed in the silica gel layer.
- the Rf value was determined in the same manner as in Example 2, the Rf value of the raffinate component of benzoin ethyl ether was 0.57, and the Rf value of the extract component was 0.30.
- Example 5 Example of optical resolution of Hexobarbital Approximately 1 ⁇ L of 1% ethyl acetate solution of Hexobarbital is spotted on the TLC plate prepared in Example 1, and a mixture containing n-hexane and isopropanol at a volume ratio of 9: 1. Development was performed in the same manner as in Example 2 except that the solvent was used as the developing solution in the first development and isopropanol was used as the developing solution in the second development.
- the TLC plate was dried with cold air, irradiated with ultraviolet rays, and the positions of the raffinate component and extract component spots were confirmed in the silica gel layer.
- the Rf value was determined in the same manner as in Example 2, the Rf value of the raffinate component of Hexobarbital was 0.33, and the Rf value of the extract component was 0.21.
- Example 6 Example of optical resolution of treger base About 1 ⁇ L of 1% ethyl acetate solution of treger base (TB) was spotted on the TLC plate prepared in Example 1, and developed in the same manner as in Example 2. .
- the TLC plate was dried with cold air, irradiated with ultraviolet rays, and the positions of the raffinate component and extract component spots were confirmed in the silica gel layer.
- the Rf value was determined in the same manner as in Example 2, the Rf value of the raffinate component of TB was 0.56, and the Rf value of the extract component was 0.41.
- the sample is developed in the longitudinal direction of the TLC plate, the distance W1 from the spotting position to the tip of the spot in the longitudinal direction of the TLC plate, and And the distance W2 from the spotting position in the direction orthogonal to the longitudinal direction to the spot tip was measured, and the ratio of W2 to W1 (W2 / W1) was determined.
- W2 the spotting position was set to zero, the second separating agent layer side was positive, and the opposite side of the TLC plate in the first separating agent layer was negative. Since the spots of sudan and p-aminoazobenzene originally have unique colors, the spot detection in the first separating agent layer was performed by visual observation. The results are shown in Table 1. From Table 1, it became clear that W2 / W1 can be controlled by changing the addition amount of gypsum and CMC1110.
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Abstract
Description
(第一分離剤層の調整)
ダイセル化学工業株式会社製CHIRALPAK IA(同社の登録商標)の充填剤(「IA充填剤」とも言う)6gと、石膏0.89gと、2%CMC(カルボキシメチルセルロース)1110(ダイセル化学工業株式会社製)水溶液6gとを、水3.03g、エタノール3.01gの混合溶液に添加し、超音波を照射しながら十分に攪拌してスラリーを得た。
シリカゲル10.01g(ダイソー株式会社製液体クロマトグラフィー用、IR-60-5/20-U)、石膏0.50g、2%CMC(カルボキシメチルセルロース)1110(ダイセル化学工業株式会社製)水溶液14.99g、マンガン含有ケイ酸亜鉛0.1gとを、水4.79g、エタノール6.51gの混合溶液に添加し、超音波を照射しながら十分に攪拌してスラリーを得た。
これらのスラリーを、仕切り板を装着したTLCプレート作製用スプレッダを用いてガラス板の表面に均一に塗布し、図1におけるA1が2cm、A2が2.5cm、Bが10cm、厚さがそれぞれ150μmの分離剤層を形成した。なお、IA充填剤の平均粒径は20μmである。また、シリカゲルの平均粒径は14.4μmである。
スラリーを塗布したTLCプレートを風乾し、真空ポンプで引きながら60℃で3時間真空乾燥し、IA充填剤の層を第一の分離剤層とし、前記シリカゲルの層を第二の分離剤層とするTLCプレートを得た。
トランス-スチルベンゼンオキシド(t-SO)の1%の酢酸エチル溶液の約1μLを、実施例1で作製したTLCプレートの長手方向を縦とした時の下から約1.5cmの位置に点着した。n-ヘキサンとエタノールを体積比で9:1で含有する混合溶剤を収容した展開槽内に、試料のスポットを下にしてTLCプレートを収容し、IA分離剤層の長手方向に試料を展開させた。
フラバノン(FLV)の1%の酢酸エチル溶液の約1μLを実施例1で作製したTLCプレートに点着し、実施例2の方法と同じ方法で展開した。
ベンゾインエチルエーテルの1%の酢酸エチル溶液の約1μLを実施例1で作製したTLCプレートに点着し、n-ヘキサンと酢酸エチルを体積比で9:1で含有する混合溶剤を第一の展開における展開液に用い、酢酸エチルを第二の展開における展開液に用いた以外は、実施例2の方法と同じ方法で展開した。
Hexobarbitalの1%の酢酸エチル溶液の約1μLを実施例1で作製したTLCプレートに点着し、n-ヘキサンとイソプロパノールを体積比で9:1で含有する混合溶剤を第一の展開における展開液に用い、イソプロパノールを第二の展開における展開液に用いた以外は、実施例2の方法と同じ方法で展開した。
トレガーベース(TB)の1%の酢酸エチル溶液の約1μLを実施例1で作製したTLCプレートに点着し、実施例2の方法と同じ方法で展開した。
ダイセル化学工業株式会社製CHIRALPAK AD(同社の登録商標)の充填剤(「AD充填剤」とも言う)及び前記シリカゲル100質量部に対して下記表1に記載の質量部数で石膏及びCMC1110を用いて、バインダの添加量を変えたTLCプレート1~6を作製し、作製したTLCプレート1~6を用い、スダンI及びp-アミノアゾベンゼンの各試料の1%の酢酸エチル溶液の約1μLを、TLCプレートの長手方向を縦とした時の下から約1.5cmの位置に点着し、アセトニトリルを収容した展開槽内に、試料のスポットを下にしてTLCプレートを収容し、TLCプレートの長手方向に試料を展開させ、TLCプレートの長手方向における点着位置からスポットの先端までの距離W1、及び、前記長手方向に直交する方向における点着位置から前記スポット先端までの距離W2を測定し、W1に対するW2の比(W2/W1)を求めた。W2は、点着位置をゼロとし、第二の分離剤層側を正、その反対の、第一の分離剤層におけるTLCプレートの端縁側を負とした。スダン及びp-アミノアゾベンゼンのスポットは元来固有の色を有することから、第一の分離剤層におけるスポットの検出は、肉眼での観察によって行った。結果を表1に示す。表1から、石膏及びCMC1110の添加量を変えることで、W2/W1がコントロールできるが明らかとなった。
2 第二の分離剤層
3 試料溶液のスポット
E エクストラクト成分のスポット
R ラフィネート成分のスポット
X 本発明における第一の方向を示す矢印
Y 本発明における第二の方向を示す矢印
Claims (7)
- 第一の分離剤層とこれに隣接する第二の分離剤層とを有する薄層クロマトグラフィープレートを用いて、紫外線又は発色試薬に対する光学応答性を有する試料中の成分を検出する方法であって、
第一の分離剤層において、第一及び第二の分離剤層の境界に沿う方向へ試料を展開させる工程と、
少なくとも第二の分離剤層の移動相を乾燥させる工程と、
前記薄層クロマトグラフィープレートの向きを変えて第一の分離剤層におけるスポットを第二の分離剤層に展開させる工程と、
第二の分離剤層に移動したスポットを紫外線の照射又は発色試薬の発色処理によって検出する工程と、を含み、
第一の分離剤層は、前記光学応答性を有する第一の分離剤で形成され、
第二の分離剤層は、第一の分離剤とは異なる前記光学応答性を有する第二の分離剤で形成され、
前記成分の光学応答性が、第一の分離剤の光学応答性と同じであることを特徴とする試料の検出方法。 - 第一の分離剤が光学異性体用分離剤であることを特徴とする請求項1に記載の試料の検出方法。
- 光学異性体用分離剤が、多糖と多糖の水酸基或いはアミノ基の一部又は全部と置き換わった芳香族エステル基、芳香族カルバモイル基、芳香族エーテル基、及びカルボニル基のいずれかとからなる多糖誘導体を含むことを特徴とする請求項2に記載の試料の検出方法。
- 第一の方向を展開方向とする第一の分離剤層と、第一の方向に直交する第二の方向において第一の分離剤層に隣接する第二の分離剤層とを有する薄層クロマトグラフィープレートにおいて、
第一の分離剤層が、紫外線又は発色試薬に対する光学応答性を有する第一の分離剤で形成され、
第二の分離剤層が、第一の分離剤とは異なる前記光学応答性を有する第二の分離剤で形成され、
第一の分離剤層で第一の方向にスポットを展開させたときに、第一の分離剤層での第一の方向におけるスポットの移動距離W1に対する、第一の分離剤層での第二の方向におけるスポットの移動距離W2の比が±0.5以下となることを特徴とする薄層クロマトグラフィープレート。 - 第一の分離剤が光学異性体用分離剤であることを特徴とする請求項4に記載の薄層クロマトグラフィープレート。
- 光学異性体用分離剤が、多糖と多糖の水酸基或いはアミノ基の一部又は全部と置き換わった芳香族エステル基、芳香族カルバモイル基、芳香族エーテル基、及びカルボニル基のいずれかを含むことを特徴とする請求項5に記載の薄層クロマトグラフィープレート。
- 第一の分離剤層とこれに隣接する第二の分離剤層とを有する薄層クロマトグラフィープレートを製造する方法であって、
紫外線又は発色試薬に対する光学応答性を有する第一の分離剤と塗布用溶剤とを含有する第一のスラリーを支持体に塗布して第一の分離剤層を形成する工程と、第一の分離剤とは異なる前記光学応答性を有する第二の分離剤と塗布用溶剤とを含有する第二のスラリーを支持体に塗布して第一の分離剤層に隣接する第二の分離剤層を形成する工程とを含み、
第一及び第二のスラリーにおいて、同じ種類の塗布用溶剤を用いる、薄層クロマトグラフィープレートの製造方法。
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CN103148997A (zh) * | 2012-12-21 | 2013-06-12 | 中国飞行试验研究院 | 航空煤油显色剂及其显色方法以及该显色剂制造方法 |
WO2013105572A1 (ja) * | 2012-01-11 | 2013-07-18 | 株式会社ダイセル | クロマトグラフ媒体 |
WO2013115350A1 (ja) * | 2012-02-03 | 2013-08-08 | 株式会社ダイセル | クロマトグラフ媒体 |
US20140248196A1 (en) * | 2011-10-05 | 2014-09-04 | Daicel Corporation | Thin-layer chromatography plate |
WO2018123476A1 (ja) * | 2016-12-26 | 2018-07-05 | パナソニックIpマネジメント株式会社 | 薄層クロマトグラフィープレート及びそれを用いた試料の分析方法 |
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WO2019102802A1 (ja) * | 2017-11-24 | 2019-05-31 | キヤノン株式会社 | クロマトグラフィー用のキット |
JP2021501889A (ja) * | 2018-06-11 | 2021-01-21 | エルジー・ケム・リミテッド | 薄膜クロマトグラフィーを利用したアルデヒド及びケトンの検出方法 |
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GB201719905D0 (en) * | 2017-11-30 | 2018-01-17 | Ucl Business Plc | Method and system for assessing analyte concentrations in samples |
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JPWO2013105572A1 (ja) * | 2012-01-11 | 2015-05-11 | 株式会社ダイセル | クロマトグラフ媒体 |
EP2803995A4 (en) * | 2012-01-11 | 2015-10-14 | Daicel Corp | CHROMATOGRAPHIC ENVIRONMENT |
WO2013105572A1 (ja) * | 2012-01-11 | 2013-07-18 | 株式会社ダイセル | クロマトグラフ媒体 |
US9726649B2 (en) | 2012-01-11 | 2017-08-08 | Daicel Corporation | Chromatographic medium |
WO2013115350A1 (ja) * | 2012-02-03 | 2013-08-08 | 株式会社ダイセル | クロマトグラフ媒体 |
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WO2019102802A1 (ja) * | 2017-11-24 | 2019-05-31 | キヤノン株式会社 | クロマトグラフィー用のキット |
CN108459133A (zh) * | 2018-04-12 | 2018-08-28 | 成都爱恩通医药科技有限公司 | 薄层色谱分析自动化智能检测仪器及其实时在线检测方法 |
CN108459133B (zh) * | 2018-04-12 | 2024-04-23 | 成都爱恩通医药科技有限公司 | 薄层色谱分析自动化智能检测仪器及其实时在线检测方法 |
JP2021501889A (ja) * | 2018-06-11 | 2021-01-21 | エルジー・ケム・リミテッド | 薄膜クロマトグラフィーを利用したアルデヒド及びケトンの検出方法 |
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US11604176B2 (en) | 2018-06-11 | 2023-03-14 | Lg Chem, Ltd. | Method for detecting aldehyde and ketone by using thin layer chromatography |
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Also Published As
Publication number | Publication date |
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CN103026224A (zh) | 2013-04-03 |
JPWO2011149041A1 (ja) | 2013-07-25 |
EP2579034A4 (en) | 2014-07-16 |
US9234881B2 (en) | 2016-01-12 |
CN103026224B (zh) | 2015-07-22 |
EP2579034B1 (en) | 2016-11-02 |
JP5941405B2 (ja) | 2016-06-29 |
US20130067996A1 (en) | 2013-03-21 |
EP2579034A1 (en) | 2013-04-10 |
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