JPH10185894A - Preparation of filler for high-performance liquid chromatography - Google Patents

Abstract

PROBLEM TO BE SOLVED: To make a liquid chromatograph to sufficiently display a separating performance which a filler has, by adopting a nitrogen flash drying method in order to dry and remove a coating solvent. SOLUTION: As a substance, with which a carried is coated and which has a separating function, it is preferable to use a polysaccharide derivative. As the kind of the polysaccharide derivative, an ester derivative, a carbonate derivative or the like is enumerated, but an aromatic carbonate derivative is preferable. On the other hand, as a carrier, it is preferable to use an inorganic carrier, but it is especially preferable to use silica gel. Then, in a coating operation, a polysaccharide aromatic carbonate derivative is dissolved in a solvent so as to be used as a dope, and it is dropped slowly while it is stirred in the carrier, and the carrier is coated uniformly. At this time, in order to remove the coating solvent after the coating operation, a nitrogen flash drying method is used, and the solvent is heated under a nitrogen flash so as to be dried and removed. In this manner, it is possible to prepare a filler which sufficiently displays a separating performance.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing a packing material for high performance liquid chromatography, and more particularly to a method for producing a packing material having excellent separation performance in the field of optical isomer separation. is there.

[0002]

2. Description of the Related Art As a packing material for high performance liquid chromatography, a substance having a separating function in the form of particles itself (crushed, beaded, etc.) is used as a packing material. There are types such as those bonded and coated on a carrier. In particular, in the field of optical isomer separation, a substance obtained by coating an optically active substance having an optical isomer separation function on a carrier is actually often used. For example, a filler in which an optically active crown ether compound as a low molecular weight compound is coated on a carrier (Japanese Patent Application Laid-Open No. 62-21053), an optically active triphenylmethyl methacrylate polymer as a synthetic polymer Filler coated on a carrier (JP-A-57-150,4
No. 32), a filler obtained by coating cellulose triacetate which is a polysaccharide derivative on a carrier (Japanese Patent Laid-Open No. 60-8282,
No. 858), a filler in which cellulose benzoate is coated on a carrier (Japanese Patent Application Laid-Open No. 60-040,952), a filler in which cellulose phenylcarbamate is coated on a carrier ( Japanese Patent Laid-Open No. 60-108,751) and the like are known, and are commercially available due to their high optical resolution and are widely used.

[0003] However, in columns packed with these packing materials, separation is often insufficient depending on the racemic compound to be separated. In such a case, countermeasures have been taken by, for example, performing separation by changing the type of the substance having the separation function. In view of such problems, the problem to be solved by the present invention is to provide a high performance liquid chromatography in which a substance having a separation function is coated on a carrier.
It is an object of the present invention to provide a method for producing a filler capable of sufficiently exhibiting its performance without changing the type of a substance having a separating function in the method for producing a filler for use.

[0004]

Means for Solving the Problems As a result of intensive studies, the present inventor has found that a method for producing a packing material for high performance liquid chromatography in which a substance having a separating function is coated on a carrier, after the coating, Instead of the conventional vacuum drying method (vacuum drying method) as a method for drying and removing the coating solvent,
By adopting the nitrogen flash drying method, it has been found that the separation performance of the filler is sufficiently exhibited, and the present invention has been completed.

That is, the present invention relates to a method for producing a filler comprising a substance having a separating function coated on a carrier,
A method for producing a packing for high-performance liquid chromatography, comprising drying and removing a coating solvent by a nitrogen gas drying method after coating.

A conventional method of removing a coating solvent by drying after coating is disclosed in the above-mentioned Japanese Patent Application Laid-Open No. 60-0828.
No. 58 (filler in which cellulose triacetate is coated on a carrier) and JP-A-60-040,952 (filler in which cellulose benzoate is coated on a carrier); As specifically disclosed in Examples of JP-A-60-108,751 (filler obtained by coating cellulose carbamate on a carrier), a vacuum drying method (vacuum drying method) is used. Was. The vacuum drying method is a method of heating under reduced pressure to evaporate and dry the coating solvent.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, a substance having a separating function may be any substance capable of separating a liquid mixture, and may be an optically active triphenylmethyl methacrylate polymer, an optically active (meth ) Acrylic amide polymers, polysaccharide derivatives and the like are preferred, and polysaccharide derivatives are preferred.

[0008] The polysaccharide used as the raw material of the polysaccharide derivative may be any one of a synthetic polysaccharide, a natural polysaccharide, and a natural modified polysaccharide.
Any substance may be used as long as it is optically active, but those having a high binding mode regularity are preferred. For example, β-1,4-
Glucan (cellulose), α-1,4-glucan (amylose, amylopectin), α-1,6-glucan, β
-1,4-galactan, β-1,6-glucan (pusulan), β-1,3-glucan (for example, cardan, schizophyllan), α-1,3-glucan, β-1,4-mannan, α-1,6-mannan, β-1,2-fructan (inulin), β-2,6-fructan (levan),
β-1,4-xylan, β-1,4-chitosan, β-
1,4-N-acetylchitosan (chitin), pullulan,
These include agarose and alginic acid, and also include amylose-containing starch and cyclodextrin, which is a cyclic polysaccharide. Particularly preferred are β-1,4-glucan (cellulose) and α-1,4-, from which high-purity polysaccharides can be easily obtained.
Glucan (amylose, amylopectin), β-1,4
-Chitosan, β-1,4-N-acetylchitosan (chitin), β-1,4-mannan, β-1,4-xylan,
Inulin, cardan, cyclodextrin and the like.
The number average degree of polymerization of these polysaccharides (the average number of pyranoth rings or furanoth rings contained in one molecule) is 2 or more, preferably 5 or more, and there is no particular upper limit, but it is 500 or less. Is preferred for ease of handling.

Examples of the type of polysaccharide derivative used in the present invention include ester derivatives, carbamate derivatives, ether derivatives and the like. Preferred are carbamate derivatives, and more preferred are aromatic carbamate derivatives. -Derivative.

The aromatic carbamate derivative of the polysaccharide used in the present invention is an average of three hydroxyl groups of the polysaccharide.
0 to 100%, preferably 85 to 100% on average, is substituted with a group represented by the following formula (I).

[0011]

Embedded image (Wherein, R ^{1 to} R ⁵ are a hydrogen atom, an alkyl group having 1 to 8 carbon atoms, an alkoxy group having 1 to 8 carbon atoms, and 6 to 14 carbon atoms.
Or a halogen atom. In order to synthesize the aromatic carbamate derivative of this polysaccharide,
The usual reaction for producing urethane from alcohol and isocyanate can be applied as it is. For example, it can be obtained by reacting a corresponding isocyanate with a polysaccharide in a suitable solvent using a Lewis base such as a tertiary amine or a Lewis acid such as a tin compound as a catalyst. Also,
The isocyanate can be easily synthesized, for example, by allowing phosgene to act on the amino group of the corresponding aniline derivative.

The carrier used in the present invention includes an organic carrier or an inorganic carrier, preferably an inorganic carrier.
Suitable as inorganic carriers are silica gel, alumina,
Examples include magnesia, titanium oxide, glass, silicate, kaolin and the like, and particularly preferred is silica gel. The particle size of the carrier varies depending on the size of the column used, but is generally 1 μm to 10 mm, preferably 1 μm
３００300 μm, preferably porous, with an average pore diameter of 10 Å-100 μm, preferably 50 Å-50,000.
Angstrom. The amount of the polysaccharide derivative retained on the carrier is 1 to 100% by weight, preferably 5 to 100% by weight based on the carrier.
５０50% by weight.

As a method of coating an aromatic carbamate derivative of a polysaccharide on a carrier, a dope is prepared by dissolving the aromatic carbamate derivative of a polysaccharide in a solvent, and the dope is slowly added to the carrier with stirring. Add dropwise and uniformly coat the carrier
Let me As the coating solvent, a solvent that can well dissolve the aromatic carbamate derivative of the polysaccharide is used, and specific examples include acetone, methylene chloride, dichloromethane, tetrahydrofuran, dioxane and the like.

In the present invention, the coating solvent removal method after coating uses a nitrogen flash drying method. this is,
The solvent is heated under a nitrogen stream to dry and remove the solvent. Drying conditions, that is, nitrogen flow rate, temperature, pressure, time, etc.
Since it differs depending on the type of the coating solvent, it is determined experimentally.

The method of packing the obtained packing material into a column is as follows.
An ordinary slurry filling method is used, and n-hexane, 2-propanol, methanol or the like is used as a solvent at this time.

[0016]

EXAMPLES The present invention will now be described specifically with reference to examples, but the present invention is not limited to these examples.

Example 1 A viscous solution prepared by dissolving 10 g of cellulostris (3,5-dimethylphenylcarbamate) in 60 ml of acetone.
The solution was slowly added dropwise to 40 g of aminopropylsilane-treated silica gel (manufactured by Nagel Co., Ltd., average particle diameter 7 μm, average pore diameter 1000 Å) while stirring at 25 ° C. Keep stirring at a temperature of 45
After the temperature was raised to 0 ° C., acetone as a coating solvent was removed by passing dry nitrogen gas through the system at 1 ml / min. After the dried filler was dispersed in 250 ml of methanol, the slurry was filtered to recover a solid part (powder). This solid portion was removed under reduced pressure (2 to 5 Toor.).
At 60 ° C. for 3 hours to obtain a filler. Subsequently, this filler was mixed with n-hexane / 2-propanol (= 9/1).
(Volume ratio)), a stainless steel column (inner diameter 0.46 cm, length 25 cm) by a slurry method using a mixed solvent of
Under the following conditions, and an optical resolution experiment was performed on a Trega-base racemate.

Mobile phase: n-hexane / 2-propanol (= 9/1 (volume ratio)) Flow rate: 1.0 ml / min Detection: UV detector 254 nm (sensitivity 0.16 AFU)
S) Temperature: 25 ° C. Injection amount: 5 μl (500 ppm) As a result of the optical resolution experiment, the separation coefficient (α) was 1.45.

Example 2 A packing material was obtained and packed in a column in the same manner as in Example 1 except that the drying time of the coating solvent was changed, and a tracer-based racemic optical resolution experiment was carried out. went. As a result of the optical resolution experiment, the separation coefficient (α) was 1.51.

Comparative Example 1 A viscous solution prepared by dissolving 10 g of cellulostris (3,5-dimethylphenylcarbamate) in 60 ml of acetone.
The solution was slowly added dropwise to 40 g of aminopropylsilane-treated silica gel (manufactured by Nagel Co., Ltd., average particle diameter 7 μm, average pore diameter 1000 Å) while stirring at 25 ° C. Keep stirring at a temperature of 45
After the temperature was lowered to 10 ° C., the pressure in the system was reduced (10 Torr), and acetone as a coating solvent was dried and removed. After the dried filler was dispersed in 250 ml of methanol, the slurry was filtered to recover a solid part (powder). The solid part was dried under reduced pressure (2 to 5 Toor.) At 60 ° C. for 3 hours to obtain a filler. Next, this filler was packed in a stainless steel column (inner diameter 0.46 cm, length 25 cm) by a slurry method using a mixed solvent of n-hexane / 2-propanol (= 9/1 (volume ratio)). In the same manner as in Example 1, an optical resolution experiment was carried out on a tregabase racemate. As a result of the optical resolution experiment, the separation coefficient (α) was 1.3.
It was 8.

Comparative Example 2 A packing material was obtained and packed in a column in exactly the same manner as in Comparative Example 1 except that the drying time of the coating solvent was changed. went. As a result of the optical resolution experiment, the separation coefficient (α) was 1.35.

[0022]

According to the present invention, there is provided a method for producing a packing material capable of sufficiently exhibiting separation performance in a method for producing a packing material for high performance liquid chromatography in which a substance having a separating function is coated on a carrier. We were able to.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI G01N 30/56 G01N 30/56 Z

Claims (6)

[Claims] 1. A method for producing a filler in which a substance having a separating function is coated on a carrier, wherein the coating solvent is dried and removed by a nitrogen gas drying method after the coating. A method for producing a packing material for high performance liquid chromatography. 2. The packing material for high performance liquid chromatography according to claim 1, wherein the substance having a separating function is selected from optically active triphenylmethyl methacrylate polymer, optically active (meth) acrylic amide polymer and polysaccharide derivative. Manufacturing method. 3. The method for producing a packing for high performance liquid chromatography according to claim 1, wherein the substance having a separating function is a polysaccharide derivative. 4. The polysaccharide derivative is an aromatic carbamate derivative of a polysaccharide in which 30 to 100% of the hydroxyl groups of the polysaccharide are substituted on the average by a group represented by the following formula (I). The method for producing a packing for high performance liquid chromatography according to the above. Embedded image (Wherein, R ^{1 to} R ⁵ are a hydrogen atom, an alkyl group having 1 to 8 carbon atoms, an alkoxy group having 1 to 8 carbon atoms, and 6 to 14 carbon atoms.
Or a halogen atom. ) 5. The aromatic carbamate derivative of a polysaccharide is cellulose tris (3,5-dimethylphenylcarbamate).
5. The high performance liquid chromatography according to claim 4, wherein
Manufacturing method for fillers. 6. The method according to claim 1, wherein the separation function is an optical isomer separation function.