JPH05232099A - Thin layer chromatogram - Google Patents

Abstract

PURPOSE:To provide a thin later chromatogram which can separate by means of thin later chromatography each of optical isomers from a mixture containing optical isomers. CONSTITUTION:This thin layer chromatogram is made up by being provided with a filling agent layer 11 for optical resolution on the surface of a flat platelike support body. The filling agent layer 11 for optical resolution is provided on the whole surface or the partial surface of the flat platelike support body.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a thin layer chromatogram, and more particularly to an optically resolvable thin layer chromatogram capable of efficiently separating optical isomers.

[0002]

BACKGROUND OF THE INVENTION As is well known, the optical isomers of chemically identical compounds generally have different effects on the living body. Therefore, in the fields of medicine, pharmaceuticals, biochemistry-related industries, etc., it has become a very important issue to prepare an optically pure compound for the purpose of improving the drug efficacy per unit and preventing the chemical damage due to side effects. .. In order to separate, ie, optically resolve a mixture of such optical isomers, there are conventionally known diastereomer method, crystallization method, enzyme method, separation membrane method, etc. In many cases, the types are limited, and there is a drawback that they are generally unsuitable. An object of the present invention is to provide a thin layer chromatogram capable of separating each optical isomer from a mixture containing optical isomers by thin layer chromatography.

[0003]

The invention according to claim 1 for solving the above-mentioned problems is characterized in that an optical resolution filler layer is provided on the surface of a flat plate-like support. It is a layer chromatogram, and the invention according to claim 2 is the thin layer chromatogram according to claim 1, wherein the filling layer for optical resolution is provided on the entire surface of a flat plate-like support, and the invention according to claim 3 is The invention is the thin-layer chromatogram according to claim 1, wherein the filler layer for optical resolution is provided on a part of the surface of a flat support.

The present invention will be described in more detail below. -Plate-shaped support-As the plate-shaped support in the present invention, all conventional materials can be used, and examples thereof include glass, polyester film, aluminum foil, and plastic film. Further, it is preferable that the surface of the support is clean and smooth. The size of the flat plate-shaped support is not particularly limited. The thickness of the support is not particularly limited.

-Filler for optical resolution- The filler for optical resolution is an optically active polymer compound,
And a filler using a low molecular weight compound having optical resolution. Examples of the optically active polymer compound include a filler in which a silica gel derivative is supported by a polysaccharide derivative, a polyacrylate derivative, or a polyamide derivative, or a filler in which the polymer itself is granulated without using silica gel. You can

Examples of polysaccharide derivatives include those obtained by derivatizing polysaccharides. As the above-mentioned polysaccharide,
There is no particular limitation as long as it has optical activity, and examples thereof include synthetic polysaccharides, natural polysaccharides and natural product-modified polysaccharides. Particularly preferred is one with a high degree of regularity in the binding mode.

Examples of the above-mentioned polysaccharides include β-1,4 glucan (cellulose), α-1,4-glucan (amylose and amylopectin), α-1,6-glucan (dextran) and β-1,6-. Glucan (Bustulan), β
-1,3-Dalcan (for example, curdlan, schizophyllan, etc.), α-1,3-glucan, β-1,2-glucan (Crown Gall polysaccharide), β-1,4-galactan, β-1,4 -Mannan, α-1, 6-mannan, β
-1,2-fructan (inulin), β-2,6-fructan (levan), β-1,4-xylan, β-1,3
-Xylan, β-1,4-chitosan, β-1,4-N-
Acetyl chitosan (chitin), pullulan, agarose,
Examples thereof include alginic acid and starch containing amylose.

Among them, particularly preferable ones are cellulose, amylose, β-1,4-chitosan, chitin, β-1,4-mannan, β-1,4-, from which high-purity polysaccharide can be easily obtained. Xylan, inulin, cardran and the like. The number average degree of polymerization of these polysaccharides (the average number of pyranose or furanose rings contained in one molecule) is preferably 2 or more, and particularly preferably 5 or more. The upper limit of the number average degree of polymerization of the polysaccharide is not particularly limited, but it is preferably 500 or less in consideration of easy handling.

Examples of the method for derivatizing the above-mentioned polysaccharide include esterification, etherification, carbamate formation and the like. Examples of the low molecular weight compound having optical resolution include crown ether or its derivative and cyclodextrin or its derivative. These low molecular weight compounds are usually used by supporting them on a carrier such as silica gel.

The optical resolution filler may be a commercially available product, for example, CH manufactured by Daicel Chemical Industries, Ltd.
IRALCEL OB (registered trademark), CHIRALCEL OD (registered trademark),
CROWNPAK CR (+) (registered trademark), CHIRALCEL CA-1 (registered trademark), CHIRALCEL OA (registered trademark), CHIRALCEL OK (registered trademark), CHIRALCEL OJ (registered trademark), CHIRALCEL OC (registered trademark), CHIRALCEL OF ( Registered trademark), CHIRALCEL OG
(Registered trademark), CHIRALPAK WH (registered trademark), CHIRALPAK
WM (registered trademark), CHIRALPAK WE (registered trademark), CHIRALPA
Preferable examples include K OT (+) (registered trademark), CHIRALPAK OP (+) (registered trademark), CHIRALPAK AS (registered trademark), and CHIRALPAK AD (registered trademark).

The average particle size of the filler varies depending on the type of optical isomer to be optically resolved and the type of eluent described later, but is usually 0.1 to 100 μm, preferably 1 to 75 μm. Is.

-Formation of Filler Layer for Optical Resolution- In the present invention, the filler layer for optical resolution is provided on the entire surface or a part of the surface of the flat plate-shaped support. The filler layer for optical resolution is preferably provided on the entire surface or a part of the surface of the plate-shaped support, depending on the detection method, the isomer to be optically resolved, the characteristics of the filler for optical resolution, and the like.

In particular, in the case of using a fluorescence detection method which is commonly used in the detection of thin layer chromatography, the isomers to be optically resolved and the difference in the ultraviolet absorption band of the packing for optical resolution are used. Therefore, it is preferable to provide the filler layer for optical resolution on the entire surface or a part of the surface. For example, when the filler for optical resolution does not emit fluorescence, or even when it emits fluorescence, it has an ultraviolet absorption band different from the ultraviolet absorption band of the isomer to be optically resolved, as described above, A filler layer for optical resolution may be provided on the entire surface of the flat plate-shaped support.

However, when the filler for optical resolution has the same ultraviolet absorption band as the ultraviolet absorption band of the isomer to be optically resolved, when ultraviolet rays are irradiated at the time of detection,
Not only the optically resolved isomer spots but also the optically resolving filler layer emits fluorescence. Therefore, if spots of the desired isomers are present on the filler layer for optical resolution, these isomers cannot be detected by fluorescence. Therefore, it is necessary to carry out the detection by making the optically resolved fluorescing spot of the isomer exist on the filler layer which does not fluoresce or has a different ultraviolet absorption band.

Therefore, in the above-mentioned case, a filler layer for optical resolution is provided on a part of the surface of the flat plate-like support, and the other surface is filled with another filler layer such as a usual filler. An agent layer is provided. That is, in order to optically resolve the mixture of isomers in the filler layer for optical resolution and to detect the optically resolved isomers in the usual filler layer,
The filler for optical resolution is provided on a part of the surface of the support on the flat plate.

In the above-mentioned filler for optical resolution, examples of fluorescent materials that emit fluorescence by ultraviolet rays include fluorescent indicators such as magnesium tungstate or zinc silicate containing manganese. Therefore, when these fillers for optical resolution are used, the filler layer for optical resolution is provided after considering whether or not the ultraviolet absorption band is the same as the ultraviolet absorption band of the substance for optical resolution. It is preferably formed.

The shape of the partial surface is not particularly limited as long as the object of the present invention is not impaired. Also, the size of the area of the partial surface is not particularly limited as long as the object of the present invention is not impaired.

For example, as shown in FIG. 1, the surface of the lower half of the support may be the filler layer for optical resolution, and the surface of the upper half may be the usual filler layer. Further, as shown in FIG. 2, the surface of the support on which the filler layer for optical resolution is formed may have a triangular shape. Further, as shown in FIG. 3, the surface of the support on which the optical division filler layer is formed may have a circular shape.

For the optical resolution filler layer, a suspension obtained by mixing the optical resolution filler with water or a solvent is drawn on the flat plate-shaped support, and then the water and It is formed by evaporating the solvent. The solvent may be selected according to the filler for optical resolution to be used, but usually, suitable solvents are water, alcohols,
Aliphatic hydrocarbons, halogenated solvents, aromatic hydrocarbons, ethers and the like can be mentioned. Among them, hexene, heptane and carbon tetrachloride are particularly preferable.

The above-mentioned solvent may be used not only in one kind but also in a mixture of two or more kinds, and the ratio thereof can be arbitrarily selected. For example, hexane-carbon tetrachloride (1: 2) is preferably used. To be done. In the present invention, the suspension may optionally contain a binder. Examples of the binder include synthetic polymers such as acrylic acid ester.

As a method for drawing the suspension on the surface of the plate-shaped support, a method using a spreader is usually used,
A known method such as a spraying method can be used. The spreader is a device that has a gap corresponding to the thickness of the layer to be formed and through which the suspension flows out, and may be a commercially available product or a self-made product.

As a method of using the spreader, there are a method of fixing the support and moving the spreader, a method of fixing the spreader and moving the support. The above-mentioned spraying method is a method in which the suspension is sprayed onto the support, which is suitable when the support is small.

After the suspension is drawn on the support by the above method, the water and the solvent in the suspension are evaporated. Examples of the method for evaporating water and the solvent include methods such as air drying and heat drying. However, when a volatile solvent is used, the target optical resolution filler layer is formed by sufficiently air drying. can do.

The thickness of the filler layer for optical resolution in the present application is usually preferably 100 μm to 5 mm, and particularly 100
μm to 1 mm is preferable. If the thickness of the filler layer for optical resolution is larger than 5 mm, a large amount of sample is required, and 1
If it is smaller than 00 μm, it becomes difficult to detect the sample. In the case where the filler layer for optical resolution is provided on a part of the surface of the support on the flat plate, the remaining surface of the support on the flat plate has a non-fluorescent filler layer, or an isomer for optical resolution. It is preferable to provide a filler layer having an ultraviolet absorption band different from the ultraviolet absorption band of the body.

[0025] The filler layer having no fluorescence, or
As the filler layer having an ultraviolet absorption band different from the ultraviolet absorption band of the substance to be optically resolved, commonly used silica gel, alumina, silicates such as magnesium silicate, calcium sulfate, activated carbon, polyamide, urea, cernose, etc. Fillers of can be used. The method for forming the filler layer having no fluorescence, or the filler layer having an ultraviolet absorption band different from the ultraviolet absorption band of the isomer to be optically resolved is the same as the method for forming the filler layer for optical resolution described above. Can be formed.

-Optical isomers which can be optically resolved by the thin layer chromatogram of the present invention-The optical isomers which can be optically resolved by the thin layer chromatogram of the present invention are not particularly limited,
For example, various compounds used in the fields of medicine, agricultural chemicals, foods, feeds, flavors, etc., such as β-blockers for pharmaceuticals, EPN which is an organophosphorus pesticide, glutamic acid monosodium salt which is a chemical seasoning, and menthol which is a flavoring agent. And the like, and further, optically active alcohols, optically active esters, and the like.

-Method of Optical Resolution Using Thin Layer Chromatogram of the Present Invention-Optical resolution using the thin layer chromatogram obtained by the present invention is carried out by an ordinary thin layer chromatography method. be able to. That is, a trace amount of the mixture of optical isomers is attached to the bottom of the thin-layer chromatogram of the present invention, and one end of the thin-layer chromatogram is immersed in an eluent in a closed developing tank to obtain the optical isomer mixture. It can be optically divided depending on the developing method.

Generally, the optical isomers have different degrees of adsorption with respect to the filler for optical resolution. Therefore,
Due to this difference in the degree of adsorption, the migration distance of each isomer on the thin layer chromatogram differs. Therefore, a filler layer for optical resolution is provided in the thin layer chromatogram of the present invention so that the optical isomers can be separated by utilizing this difference in movement.

The method for optical resolution using the thin layer chromatogram of the present invention will be specifically described below.
When the mixture of these optical isomers is attached to a thin layer chromatogram, it is usually preferable to form a dilute solution of about 0.1 to 5%.

As a solution used for preparing a dilute solution, for example, alcohols such as methanol, ethanol and isopropanol, organic solvents such as hydrocarbons such as hexane, salts of an aqueous solution of copper sulfate and an aqueous solution of perchlorate are used. An aqueous solution containing can be mentioned. Which solution is preferable is appropriately determined according to the kind of compound to be subjected to optical resolution. In addition, as a solution used for forming a dilute solution, it is usually preferable to use the same solution as the eluent.

As a method for attaching the mixture of optical isomers to the thin layer chromatogram, a known method can be used, and the mixture is attached to the thin layer chromatogram of the present invention by using a capillary, a micropipette or the like. When the mixture of optical isomers is deposited, it is preferable that the spot has a diameter of about 2 mm or less and is deposited in several times so as not to spread.

The thin layer chromatogram with the attached optical isomers is immersed in the eluent. As the eluent,
Examples thereof include alcohols such as methanol, ethanol and isopropanol, organic solvents such as hydrocarbons such as hexane, and aqueous solutions containing salts such as copper sulfate aqueous solution and perchlorate aqueous solution. Which eluent is preferable is appropriately determined depending on the type of isomer to be optically resolved.

As a method of immersing in the eluent, a usual method can be used and there is no particular limitation. For example, in a sealed container, one end of the thin layer chromatogram on the side on which the optical isomer mixture is attached is soaked in the eluent so that the attached optical isomer mixture is not submerged in the eluent. Examples thereof include a method of leaning a chromatogram.

The time of immersion in the eluent is not particularly limited as long as the optical isomer can be sufficiently optically resolved. However, when using a thin-layer chromatogram in which the optical resolution filler layer is partially provided on the surface, the isomers optically resolved by the optical resolution filler layer are packed in a layer other than the optical resolution filler layer. The immersion time is appropriately selected so that it can be detected on the agent layer.

As a method for detecting the optically resolved isomer, a commonly used method can be used without any particular limitation. For example, a chemical method such as a method of detecting an optically resolved isomer by spraying a color former, a physical method such as a method of detecting by irradiating with ultraviolet rays to cause fluorescence, and further, a thin layer chromatogram is used as a medium. Biological methods such as a method of detecting by a growth inhibitory effect after contact with the.

[0036]

Industrial Applicability According to the method of the present invention, it is possible to provide a thin layer chromatogram capable of separating each optical isomer from a mixture containing optical isomers by thin layer chromatography.

[0037]

【Example】

(Example 1) Cellulose tris (3,5-dimethylphenyl _{carbamate) (CTPC-3,5-Me 2} ),
A filler for optical resolution was used which was adsorbed by about 20% by weight on a silica gel (manufactured by Fuji Davison) having a particle size of 20 μm, a pore size of 800 Å and treated with diphenyldichloromethane.

9.5 g of this optical resolution filler was mixed with 75 ml of a hexane-carbon tetrachloride solution having a mixing ratio of 1: 2, and the resulting suspension was placed on a glass slide of 2.5 × 7.5 cm. A thin layer chromatogram was prepared by coating. Using the thin-layer chromatogram obtained by the above method, the optical resolution of the racemic anthracene derivative as shown in the figure below was performed.

[0039]

[Chemical 1]

Hexane-2-propanol having a mixing ratio of 90:10 was used as an eluent. It took about two and a half minutes for the eluent to reach near the top of the thin layer chromatogram. This development was repeated 3 times. After the development, the thin layer chromatogram was irradiated with an ordinary ultraviolet lamp, and the result as shown in FIG. 4 was obtained. As shown in FIG. 4, two completely separated fluorescent spots appeared on the optical resolution filler layer, and each isomer could be optically resolved from the racemate.

[Brief description of drawings]

FIG. 1 is an explanatory view showing an example of a thin layer chromatogram of the present invention.

FIG. 2 is an explanatory view showing an example of the thin layer chromatogram of the present invention.

FIG. 3 is an explanatory view showing an example of the thin layer chromatogram of the present invention.

FIG. 4 is an explanatory view showing the result of one experiment of optical resolution using the thin layer chromatogram of the present invention.

[Explanation of symbols]

11 Packing Layer for Optical Resolution in Thin Layer Chromatogram 12 Silica Gel Layer in Thin Layer Chromatogram 20 When a mixture of optical isomers is attached 21 Fluorescent spot of optically resolved optical isomers 22 Optically resolved optical isomers Fluorescent spot

Claims (3)

[Claims] 1. A thin-layer chromatogram, which comprises a flat support and an optical resolution filler layer provided on the surface of the support. 2. The thin layer chromatogram according to claim 1, wherein the filling layer for optical resolution is provided on the entire surface of a flat plate-shaped support. 3. The thin layer chromatogram according to claim 1, wherein the optical resolution packing layer is provided on a part of the surface of a flat plate-shaped support.